Methods of treating sarcoidosis with TNF alpha antibodies and related compositions and methods

Abstract

Compositions and related methods are provided, the compositions comprising an anti-TNF alpha antibody sialylated with one or more N-acetylneuraminic acid molecules. In some embodiments, methods for treating or preventing sarcoidosis and / or granulomegaly or formation in a patient in need thereof are provided.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 431,607, filed on December 9, 2022, and U.S. Provisional Patent Application No. 63 / 487,060, filed on February 27, 2023. The content of each of these provisional applications is incorporated by reference in its entirety.

[0003] Sequence Listing Incorporated by Reference

[0004] This application contains an ST.26 compliant sequence listing, which is submitted in parallel in xml format via the Patent Center and is hereby incorporated by reference in its entirety. The.xml copy created on December 7, 2023 is named 140505-8004WO00_SL.xml and has a size of 0.026MB (25.2KB) (25,856 bytes). Background Art

[0005] Sarcoidosis is a chronic multisystem inflammatory disorder of unknown etiology, characterized by the presence of non-caseating epithelioid granulomas with mononuclear cell infiltration and microarchitectural disruption. This rare disease affects the skin, eyes, heart, and central nervous system, and >90% of cases involve the lungs. Subjects with pulmonary manifestations may progress to loss of lung function, and immunomodulatory therapies may be beneficial in these cases. Current treatment options are suboptimal due to adverse side effects, limited efficacy, or unavailability of the most effective treatments.

[0006] Macrophage-derived tumor necrosis factor alpha (TNFα) is involved in the induction and maintenance of granulomas, and high levels of TNFα (and high levels of TNFα released from alveolar macrophages) appear to be associated with disease progression. In chronic sarcoidosis, preventing granuloma formation and limiting tissue damage and fibrosis are the goals of therapy. In some cases, these goals can be achieved by targeted immunosuppression of pro-inflammatory cytokines such as TNFα. Infliximab is a monoclonal antibody (mAb) that targets TNFα and is used to treat sarcoidosis. In a randomized controlled study of subjects with chronic refractory pulmonary sarcoidosis, low-dose and high-dose intravenous (IV) infusions of infliximab increased predicted forced vital capacity by 2.5% at 24 weeks. Notably, other TNF inhibitors have shown no efficacy for the treatment of sarcoidosis or may paradoxically cause sarcoid-like drug reactions.

[0007] First-line agents include corticosteroids (which are tapered over time), followed by second- and third-line immunomodulators, which are carefully selected to mitigate side effects, particularly those associated with systemic steroid exposure. Although corticosteroids are effective in resolving the granulomatous inflammation of sarcoidosis, up to 20% of subjects treated with systemic steroid therapy continue to exhibit a persistent granulomatous inflammatory process and progress to irreversible tissue remodeling and fibrosis.

[0008] In subjects with advanced or severe refractory sarcoidosis who are unresponsive to conventional immunosuppressive therapy, the use of TNFα inhibitors is recommended. However, pharmacokinetic analysis of the use of TNFα inhibitors in sarcoidosis is limited. Therefore, there remains a need for dose ranging to determine a dosage regimen that is safe and effective for the sarcoidosis population. SUMMARY OF THE INVENTION

[0009] In some aspects, the technology of the present invention includes a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control treatment, the method comprising administering to the subject a first dose of a composition at about 0.5 mg / kg to about 6.0 mg / kg about once every four weeks for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[0010] In some aspects, the technology of the present invention includes a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control treatment, the method comprising administering to the subject a first dose of a composition at about 0.5 mg / kg to about 6.0 mg / kg about once every two weeks for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[0011] In some aspects, the technology of the present invention includes a method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about 0.5 mg / kg to about 6.0 mg / kg about once every four weeks for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[0012] In some aspects, the technology of the present invention includes a method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about 0.5 mg / kg to about 6.0 mg / kg about once every two weeks for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[0013] In some aspects, the technology of the present invention includes a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every four weeks for a first period of time; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks for a second period of time.

[0014] In some aspects, the technology of the present invention includes a method for reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every four weeks for a first period of time; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks for a second period of time.

[0015] In some aspects, the technology of the present invention includes a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every four weeks for a first period of time.

[0016] In some aspects, the technology of the present invention includes a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every two weeks for a first period of time.

[0017] In some aspects, the technology of the present invention includes a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method for reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every four weeks for a first period of time.

[0018] In some aspects, the technology of the present invention includes a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, and the composition is used in a method of reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg approximately once every two weeks for a first time period.

[0019] In some embodiments, the first time period is about 6 weeks to about 24 weeks.

[0020] In some embodiments, the first dose is about 2 mg / kg of the composition.

[0021] In some embodiments, the first dose is about 4 mg / kg of the composition.

[0022] In some embodiments, the method further comprises administering a second dose of the composition to the subject for a second time period.

[0023] In some embodiments, the second time period is about 6 weeks to about 16 weeks.

[0024] In some embodiments, wherein the second dose comprises about 1 mg / kg to about 5 mg / kg.

[0025] In some embodiments, the second dose is about twice the first dose.

[0026] In some embodiments, the administration frequency of the second dose comprises the same administration frequency as that of the first dose.

[0027] In some embodiments, the first dose and the second dose are each administered approximately once every four weeks.

[0028] In some embodiments, the second dose is a dose substantially the same as the first dose.

[0029] In some embodiments, the first dose and the second dose are about 2 mg / kg.

[0030] In some embodiments, the administration frequency of the second dose is about twice the administration frequency of the first dose.

[0031] In some embodiments, the first dose is administered approximately once every four weeks, and the second dose is administered approximately once every two weeks.

[0032] In some embodiments, the anti-TNFα antibody comprises an asparagine-linked glycosylation site located on at least one heavy chain in the constant Fc region.

[0033] In some embodiments, the anti-TNFα antibody is sialylated at Asn 297.

[0034] In some embodiments, the anti-TNFα antibody does not contain N-glycolylneuraminic acid (Neu5Gc).

[0035] In some embodiments, the anti-TNFα antibody is an immunoglobulin G (IgG) antibody.

[0036] In some embodiments, the IgG antibody is an IgG1 antibody.

[0037] In some embodiments, the IgG1 antibody comprises a κ isotype.

[0038] In some embodiments, the anti-TNFα antibody is a chimeric antibody.

[0039] In some embodiments, the anti-TNFα antibody comprises a C-terminal lysine on at least one heavy chain.

[0040] In some embodiments, the C-terminal lysine is not trimmed.

[0041] In some embodiments, the composition is administered by infusion.

[0042] In some embodiments, the infusion is an intravenous (IV) infusion.

[0043] In some embodiments, the infusion is administered over a period of at least about 2 hours.

[0044] In some embodiments, the subject receives a cumulative dose of about 6 mg / kg, 8 mg / kg, or 16 mg / kg of the composition during the first period.

[0045] In some embodiments, the subject receives a cumulative dose of about 10 mg / kg to about 16 mg / kg of the composition during the second period.

[0046] In some embodiments, the subject receives a cumulative dose of about 28 mg / kg to about 60 mg / kg of the composition.

[0047] In some embodiments, the subject is administered at least 3, 4, 6, or 7 doses of the composition.

[0048] In some embodiments, the at least 3, 4, 6, or 7 doses are administered over a period of about 4 weeks to about 28 weeks.

[0049] In some embodiments, administration of the composition reduces the level of one or more transcripts selected from the group consisting of angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL6), interleukin 2 receptor (IL2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL1B), interleukin 12 (IL12), interleukin 18 (IL18), interferon γ (IFNG), interleukin 8 (IL8), monocyte chemoattractant protein-1 (MCP-1), chemokine (C-X-C motif) ligand 1 (CXCL1), chemokine (C-X-C motif) ligand 2 (CXCL2), chemokine (C-X-C motif) ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JunB), mitogen-activated protein kinase phosphatase 1 (MKP1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL32), intercellular adhesion molecule 1 (ICAM-1), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), and vascular cell adhesion molecule 1 (VCAM-1), relative to a control.

[0050] In some embodiments, administration of the composition reduces the levels of one or more proteins selected from the group consisting of angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL-6), interleukin 2 receptor (IL-2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL-1B), interferon γ (IFN-γ), interleukin 8 (IL-8), interleukin 12 (IL-12), interleukin 18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), chemokine (C-X-C motif) ligand 1 (CXCL1), chemokine (C-X-C motif) ligand 2 (CXCL2), chemokine (C-X-C motif) ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JUNB), mitogen-activated protein kinase phosphatase 1 (MKP-1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL-32), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1), relative to a control.

[0051] In some embodiments, administration of the composition increases one or more of the following: forced vital capacity (FVC) measurement, forced expiratory volume in 1 second (FEV1) measurement, Leicester Cough Questionnaire (LCQ) score, Short Form 36 Health Survey (SF-36) score, King's Sarcoidosis Questionnaire (KSQ) score, steroid toxicity scale (STS) score, or 6-minute walk test (6MWT) distance, relative to a control.

[0052] In some embodiments, administration of the composition reduces one or more of the following relative to a control: a patient global assessment (PGA) score, a fatigue assessment scale (FAS) score, a modified Medical Research Council (mMRC) dyspnea scale score, a Saint George's Respiratory Questionnaire (SGRQ) score, or a Borg's CR10 dyspnea score.

[0053] In some embodiments, administration of the composition reduces the need for anti-inflammatory drugs relative to a control.

[0054] In some embodiments, the subject has received or is receiving an anti-inflammatory drug.

[0055] In some embodiments, the anti-inflammatory drug is selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, efzofitimod, rituximab, adalimumab, golimumab, namilumab, and infliximab.

[0056] In some embodiments, the corticosteroid is selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone, and triamcinolone.

[0057] In some embodiments, administration of the composition reduces the granuloma size of the subject or reduces granuloma formation.

[0058] In some embodiments, the granuloma is a cutaneous granuloma, a pulmonary granuloma, a lymph node granuloma, an ocular granuloma, a hepatic granuloma, a splenic granuloma, a cardiac granuloma, or a rheumatoid granuloma.

[0059] In some embodiments, administration of the composition reduces the calcitriol level of the subject relative to a control.

[0060] In some embodiments, administration of the composition reduces the neopterin level of the subject relative to a control.

[0061] In some embodiments, administration of the composition reduces the lysozyme level of the subject relative to a control.

[0062] In some embodiments, administration of the composition reduces the soluble IL-2R (sIL-2R) level of the subject relative to a control.

[0063] In some embodiments, the subject has one or more of the following: acute sarcoidosis, chronic sarcoidosis, progressive sarcoidosis, pulmonary sarcoidosis, remitting sarcoidosis, refractory sarcoidosis, advanced sarcoidosis, ocular sarcoidosis, cutaneous sarcoidosis, or nervous system sarcoidosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] Figure 1 Shows TNFα-mediated cytotoxicity of anti-TNFα antibody XTMAB-16 relative to infliximab in A375 human melanoma cells. DP: Drug Product; TNFα: Tumor Necrosis Factor α.

[0065] Figure 2 Shows transmembrane tumor necrosis factor α (mTNFα) binding on the surface of transformed SP2 / 0 cells. Results for XTMAB-16 (squares) and infliximab (circles) are shown.

[0066] Figure 3 Is a sensorgram that shows the binding kinetics of XTMAB-16 to TNFα at different concentrations.

[0067] Figure 4 Shows the arthritis scores over time of animals treated with 20 mg / kg XTMAB-16 or 40 mg / kg XTMAB-16 relative to vehicle-treated controls. The data shown are mean ± standard error.

[0068] Figure 5A and 5B Provides a sensorgram that shows the binding kinetics of XTMAB-16 to FcγRI for batch A ( Figure 5A ) and batch B ( Figure 5B ). DS: Drug Substance; RS: Reference Standard.

[0069] Figure 6A and 6B Provides a sensorgram that shows the binding kinetics of XTMAB-16 to FcγRI for batch A ( Figure 6A ) and batch B ( Figure 6BBinding kinetics of XTMAB-16 to FcγRIIIa. DS: Drug substance; RS: Reference standard.

[0070] Figure 7A and 7B Provided are sensorgrams that show the binding kinetics of XTMAB-16 to FcγRIIb for batch A ( Figure 7A ) and batch B ( Figure 7B ). DS: Drug substance; RS: Reference standard.

[0071] Figure 8A and 8B Provided are sensorgrams that show the binding kinetics of XTMAB-16 to FcRn for batch A ( Figure 8A ) and batch B ( Figure 8B ). DS: Drug substance; RS: Reference standard.

[0072] Figure 9 Shows XTMAB-16 serum concentration over time under treatment with 10 mg / kg, 20 mg / kg, or 40 mg / kg XTMAB-16.

[0073] Figure 10 Shows the XTMAB-16 dose proportionality from the perspective of the mean maximum observed concentration (C 最大值 ).

[0074] Figure 11 Shows the XTMAB-16 dose proportionality from the perspective of the mean area under the curve (AUC (0-∞) ).

[0075] Figure 12 Shows the concentration of XTMAB-16 on day 15 after dosing on days 1, 8, and 15 in ICR mice (N = 1 to 2 / time point). ICR: Institute of Cancer Research.

[0076] Figure 13 shows the pharmacokinetic results of XTMAB-16 serum concentration over time in non-sarcoidosis subjects. The results are shown on a linear ( Figure 13A ) scale and a logarithmic ( Figure 13B ) scale. All results are presented as mean ± SD. LLOQ: Lower limit of quantification (400 ng / mL).

[0077] Figure 14 shows the pharmacokinetic results of XTMAB-16 serum concentration over time in non-sarcoidosis subjects summarized by treatment and ADA incidence. The results are shown on a linear ( Figure 14A ) scale and a logarithmic ( Figure 14B)The scale shows the results. All results are presented as mean ± SD. LLOQ: Lower Limit of Quantification (400 ng / mL).

[0078] Figure 15 Schematic illustration of the in vitro granuloma model that generates sarcoidosis. PBMC: Peripheral blood mononuclear cells.

[0079] Figure 16 Representative micrographs of the in vitro model of sarcoidosis from granuloma-like cell aggregates. PPD: Purified protein derivative; UNC: Uncoated beads.

[0080] Figure 17A - 17C Shows high responder subjects ( Figure 17A ), low responder subjects ( Figure 17B ), and subjects with sarcoidosis ( Figure 17C ) in the group analysis of granuloma formation. N = 5 / group; *p < 0.001 compared to the uncoated bead (UNC) treatment group; relative to the PPD-treated group, p < 0.01, p < 0.001, and §p < 0.05. Area fraction = percentage of the cumulative granuloma area to the total area of the image. Bar graphs are mean (μg / mL) ± SD. PPD: Purified protein derivative; PRED: Prednisone; UNC: Uncoated beads.

[0081] Figure 18A - 18C Shows the supernatant release of TNFα ( Figure 18A ), IL-1β ( Figure 18B ), and IFNγ ( Figure 18C ) in high responder subjects with sarcoidosis. N = 5 / group; *p < 0.001 compared to the uncoated bead (UNC) treatment group; relative to the PPD-treated group, p < 0.01, p < 0.001 (at the bottom, p < 0.05), and §p < 0.05. Bar graphs are mean (μg / mL) ± SD. IFN: Interferon; IL: Interleukin; PPD: Purified protein derivative; PRED: Prednisone; SD: Standard deviation; TNF: Tumor necrosis factor; UNC: Uncoated beads.

[0082] Figure 19A and 19B Show the effect of XTMAB-16 concentration on the area fraction relative to PPD ( Figure 19A ) and the effect of XTMAB-16 concentration on IL-1β level ( Figure 19B)。CI: Confidence Interval; IC50: Half Maximal Inhibitor Concentration; IL: Interleukin; PPD: Purified Protein Derivative

[0083] Figure 20A and 20B show the XTMAB-16 concentration against time on a linear scale ( Figure 20A ) and a semi-logarithmic scale ( Figure 20B ). BLQ: Below Limit of Quantification; SD: Standard Deviation; T 最大值 : Time to reach maximum concentration.

[0084] Figure 21A - 21F show the model-predicted pulmonary XTMAB-16 concentration in virtual subjects. Figure 21A and 21B show the population prediction. Figure 21C and 21D show the individual prediction. Figure 21E show the conditional weighted residuals against the population prediction. Figure 21F show the conditional weighted residual analysis against time after dosing. IC50: Concentration at half maximal MIPAR area fraction, which is defined as the percentage of the cumulative granuloma area to the total area of the image; Q2W: Every 2 weeks; Q4W: Every 4 weeks.

[0085] Figure 22 show the visual predictive check related to the prediction of the XTMAB-16 concentration over time. CL: Clearance; VPC: Visual Predictive Check.

[0086] Figure 23 show the model-predicted pulmonary XTMAB-16 concentration in a 70g individual under various dosing regimens. Q2W: Every 2 weeks; Q4W: Every 4 weeks.

[0087] Figure 24 show a high-level overview of the evaluation of XTMAB-16 in human subjects by using both a multiple ascending dose (MAD) study and a proof-of-concept (POC) study.

[0088] Figure 25 show a schematic diagram of the MAD study for evaluating XTMAB-16 in human subjects. kg: Kilogram; mg: Milligram; Q2W: Every 2 weeks; Q4W: Every four weeks.

[0089] Figure 26 show a schematic diagram of the POC study for evaluating XTMAB-16 in human subjects. Open circles represent the Q2W dosing schedule. Filled black circles represent the Q4W dosing schedule. kg: Kilogram; mg: Milligram; Q2W: Every 2 weeks; Q4W: Every four weeks; TBD: To Be Determined.

[0090] Figure 27 shows Figure 25 A schematic diagram of dose escalation in the MAD study of. kg: kilogram; mg: milligram; Q2W: every 2 weeks; Q4W: every four weeks. Detailed implementation mode

[0091] Although the technical solution of the present invention can be embodied in various forms, the following description of several embodiments is made on the understanding that the technical solution of the present invention is regarded as an illustration of the technical solution of the present invention and is not intended to limit the technical solution of the present invention to the specific embodiments shown. The titles are provided for convenience only and should not be construed as limiting the technical solution of the present invention in any way. The embodiments shown under any title can be combined with the embodiments shown under any other title.

[0092] Unless otherwise expressly indicated, the numerical values used in the various quantitative values specified in this application are represented as approximate values, as if the minimum and maximum values within the stated ranges both begin with the word "about". It should be understood that although not always explicitly stated, all numerical values are preceded by the term "about". It should be understood that such range formats are used for convenience and brevity and should be flexibly understood to include the values expressly specified as the limits of the range, as well as all individual values or sub-ranges subsumed within the stated range, as if each value and sub-range were expressly specified. For example, a ratio within the range of about 1 to about 200 should be understood to include the expressly recited limits of about 1 and about 200, but also to include individual ratios such as about 2, about 3, and about 4, as well as sub-ranges such as about 10 to about 50, about 20 to about 100, etc. It should also be understood that although not always explicitly set forth, the reagents described herein are merely exemplary and their equivalent forms are known in the art. In addition, as used herein, the term "about" when referring to measurable values such as amounts or concentrations is intended to cover variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of the specified amount.

[0093] Similarly, the scope of the technical solution of the present invention is intended to be a continuous range, including every value between the recited minimum and maximum values, as well as any range that can be formed by these values. The technical solution of the present invention includes any and all numerical values (and any range of such ratios) that can be formed by dividing the numerical values of the technical solution of the present invention by any other numerical value of the technical solution of the present invention. Therefore, those skilled in the art will understand that many such ratios, ranges, and ranges of ratios can be explicitly derived from the numerical values presented herein; and in all cases, such ratios, ranges, and ranges of ratios represent various embodiments of the technical solution of the present invention.

[0094] The terms "bind" and "binding" refer to all types of physical and chemical binding, reaction, complexation, attraction, chelation, etc.

[0095] "Amino acid" refers to a molecule containing an amino group, a carboxylic acid group, and a side chain that is unique to each amino acid. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen, and it has the general formula H2N—CHR—COOH, where R represents the side chain group. The various α-amino acids differ in the side chain moiety attached to the α-carbon. The "amino acids" of the technology of the present invention include the known naturally occurring protein amino acids, which are denoted by both the common three-letter abbreviations and the single-letter abbreviations. Generally refer to "Synthetic Peptides: A User's Guide", edited by G.A. Grant, W.H. Freeman & Co., New York (1992), including the text and tables set forth on pages 11 to 24, the teachings of which are incorporated herein by reference. As described above, the term "amino acid" also includes stereoisomers and modifications of naturally occurring protein amino acids, non-protein amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, constructs or structures designed to mimic amino acids, etc. Modified and uncommon amino acids are generally described in "Synthetic Peptides: A User's Guide", supra; Hruby et al., Biochem. J. 268:249-262 (1990); and Toniolo, Int. J. Peptide Protein Res. 35:287-300 (1990); the teachings of the entire contents of which are incorporated herein by reference.

[0096] "Antibody" refers to any form of antibody that exhibits a desired biological activity, such as inhibiting the binding of a ligand to its receptor or inhibiting the signal transduction of a ligand-induced receptor. Thus, "antibody" is used in the broadest sense and specifically encompasses, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies).

[0097] "Antibody fragment" and "antibody-binding fragment" mean antigen-binding fragments and analogs of an antibody, which typically include at least a portion of the antigen-binding region or variable region (e.g., one or more CDRs) of the parent antibody. Antibody fragments retain at least some of the binding specificity of the parent antibody. Typically, when the parent binding activity is expressed on a molar basis, the antibody fragment retains at least 10% of said activity. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the binding affinity of the parent antibody for the target. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2 and Fv fragments; bispecific antibodies; linear antibodies; single-chain antibody molecules (e.g., sc-Fv), monovalent antibodies (technology from Genmab); nanobodies (technology from Domantis); domain antibodies (technology from Ablynx); and multispecific antibodies formed from antibody fragments. Engineered antibody variants are reviewed in Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

[0098] The "Fc" region contains two heavy-chain fragments, the two heavy-chain fragments containing the C H 1 domain and C H 2 domain. The two heavy-chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the C H 3 domain.

[0099] As used herein, an antibody fragment can contain a portion of the constant region sufficient to permit heavy-chain dimerization (or multimerization) with reduced disulfide-bonding ability, e.g., in the case of such dimerization, at least one hinge cysteine that is normally involved in inter-heavy-chain disulfide bonding is altered as described herein. In another embodiment, an antibody fragment (e.g., an antibody fragment containing an Fc region) retains at least one of the biological functions normally associated with the Fc region when present in a full antibody, such as FcRn binding, antibody half-life modulation, antibody-dependent cell-mediated cytotoxicity (ADCC) function, and / or complement binding (e.g., in such case, the antibody has the glycosylation profile necessary for ADCC function or complement binding).

[0100] A "Fab fragment" contains one light chain and the C H 1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy-chain molecule.

[0101] The "Fc" region contains two heavy-chain fragments, the two heavy-chain fragments containing the C H1 domain and C H 2 domains. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the C H 3 domain.

[0102] A "Fab' fragment" contains a light chain and a heavy chain containing the V H domain and C H 1 domain and also includes a portion of the region located between the C H 1 domain and the C H 2 domain such that an interchain disulfide bond can be formed between the two heavy chains of two Fab' fragments to form an F(ab')2 molecule.

[0103] An "F(ab')2 fragment" contains two light chains and two heavy chains containing the constant region located between the C H 1 domain and the C H 2 domain such that an interchain disulfide bond is formed between the two heavy chains. Thus, an F(ab')2 fragment consists of two Fab' fragments held together by a disulfide bond located between the two heavy chains.

[0104] An "Fv region" contains the variable regions from both the heavy and light chains but lacks the constant regions.

[0105] A "single-chain Fv antibody" (or "scFv antibody") refers to an antibody fragment containing the V H domain and the V L domain of an antibody, wherein these domains are present in a single polypeptide chain. Typically, the Fv polypeptide further contains a polypeptide linker located between the V H domain and the V L domain, and the polypeptide linker enables the scFv to form the structure required for antigen binding. For a review of scFv, see Pluckthun (1994) THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, Volume 113, edited by Rosenberg and Moore, Springer-Verlag, New York, pages 269 - 315. See also International Patent Application Publication No. WO 88 / 01649 and U.S. Patents Nos. 4,946,778 and 5,260,203.

[0106] A "bispecific antibody" is a small antibody fragment having two antigen-binding sites. The fragment contains a V H -V L or V L -V H) variable light chain domain (V L ) variable heavy chain domain (V H ). By using a linker short enough to permit pairing between the two domains on the same chain, the domains are forced to pair with the complementary domain of the other chain and create two antigen-binding sites. Bifunctional antibodies are more fully described, for example, in EP 404,097; WO 93 / 11161; and Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448.

[0107] "Domain antibody fragment" is an immunologically functional immunoglobulin fragment containing only the variable region of the heavy chain or the variable region of the light chain. In some instances, two or more V H regions are covalently linked with a peptide linker to produce a bivalent domain antibody fragment. The two V H regions of the bivalent domain antibody fragment can target the same or different antigens.

[0108] The term "chimeric" antibody refers to an antibody in which part of the heavy and / or light chain is identical or homologous to the corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to the corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass and fragments of such antibodies, so long as the antibody exhibits the desired biological activity (see, e.g., U.S. Patent No. 4,816,567; and Morrison et al., 1984, Proc. Natl. Acad. Sci. USA 81:6851-6855).

[0109] The "humanized" form of a non-human (e.g., murine) antibody is a chimeric antibody that contains a minimal sequence derived from a non-human immunoglobulin. For the most part, a humanized antibody is a human immunoglobulin (recipient antibody) in which the residues of the hypervariable regions from the recipient have been replaced by the residues of the hypervariable regions from a non-human species (donor antibody) such as a mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and capacity. In some instances, the Fv framework region (FR) residues of the human immunoglobulin are replaced by the corresponding non-human residues. In addition, a humanized antibody may contain residues that are not present in the recipient antibody or the donor antibody. These modifications are made to further improve antibody performance. Generally, a humanized antibody will contain substantially all of the variable domains in at least one and typically two variable domains in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. A humanized antibody optionally will also contain at least a portion of the immunoglobulin constant region (Fc), usually at least a portion of a human immunoglobulin constant region. For additional details, see Jones et al., Nature, 321:522-525 (1986); Reichmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992).

[0110] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the techniques of the present invention may be prepared by the hybridoma method first described by Kohler et al., 1975, Nature 256:495, or may be prepared by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). For example, "monoclonal antibodies" may also be isolated from phage antibody libraries using the techniques described in Clackson et al., 1991, Nature 352:624-628 and Marks et al., 1991, J. Mol. Biol. 222:581-597. Monoclonal antibodies specifically include "chimeric" antibodies herein.

[0111] As used herein, a sequence "variant" refers to a sequence that differs from the sequence of the technology of the present invention at one or more amino acid residues but retains the biological activity of the resulting molecule.

[0112] As used herein, "percent identity" between two sequences refers to a function of the number of identical positions shared by the sequences (i.e., % homology = number of identical positions / total number of positions × 100), which function takes into account the number of gaps introduced for optimal alignment of the two sequences and the length of each gap. Comparison of sequences and determination of the percent identity between two sequences can be accomplished using mathematical algorithms. For example, the percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17 (1988)) incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Additionally, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol., 48:444-453 (1970)) algorithm of the GAP program incorporated into the GCG software package (available at www.gcg.com), using a Blossum62 matrix or a PAM250 matrix, gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0113] When referring to a ligand / receptor, antibody / antigen, or other binding pair, "specific" binding refers to a binding reaction that determines the presence of a protein (e.g., TNFα) in a heterogeneous population composed of proteins and / or other biological agents. Thus, under specified conditions, the specified ligand / antigen binds to a specific receptor / antibody and does not bind to other proteins present in the sample in significant amounts.

[0114] As used herein, a "pharmaceutically acceptable carrier" of a composition refers to a carrier or diluent that does not cause significant irritation to an organism, does not destroy the biological activity and properties of the administered active ingredient, and / or does not interact in a harmful manner with other components of the composition containing it. The term "carrier" encompasses any excipient, binder, diluent, filler, salt, buffer, solubilizer, lipid, stabilizer, or other materials known in the art for use in pharmaceutical formulations. The choice of carrier for a composition will depend on the intended route of administration of the composition. The preparation of pharmaceutically acceptable carriers and formulations containing these materials is described, for example, in Remington's Pharmaceutical Sciences, 21st Edition, Editor: University of the Sciences in Philadelphia, Lippincott, Williams & Wilkins, Philadelphia, Pa., 2005, which is incorporated herein by reference in its entirety. Some examples of physiologically acceptable carriers include antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or nonionic surfactants, such as (ICI, Inc.; Bridgewater, N.J.), polyethylene glycol (PEG), and PLURONICS TM (BASF; Florham Park, N.J.). An "excipient" of a composition refers to an inert substance added to the composition to further facilitate the administration of the compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, and various types of starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol.

[0115] As used herein, when referring to an antibody, a "control" can refer to a composition lacking an anti-TNFα antibody of the technology of the present invention or a composition having an antibody with characteristics different from those of the anti-TNFα antibody of the technology of the present invention. For example, a control can comprise an anti-TNFα antibody having an amino acid sequence or a post-translational modification state different from that of the anti-TNFα antibody of the technology of the present invention.

[0116] As used herein, when referring to a subject, a "control" can refer to a subject at baseline (i.e., before administration of a composition of the technology of the present invention or another therapeutic intervention), a subject without sarcoidosis and / or granulomas, an untreated subject, a subject administered a placebo, or a subject administered a composition different from the composition administered to the treated subject.

[0117] Anti - TNFα Antibody

[0118] Amino acid sequence

[0119] The technology of the present invention provides a composition comprising an anti-TNFα antibody or an antigen-binding fragment thereof. The anti-TNFα antibody of the technology of the present invention specifically binds to at least a portion of a TNFα protein or peptide. The TNFα protein or peptide can be produced by macrophages, monocytes, neutrophils, T cells, NK cells, or fibroblasts and can be membrane-bound or soluble. The anti-TNFα antibody can bind to human or non-human targets. In some embodiments, the anti-TNFα antibody of the technology of the present invention binds to the proteins listed in Table 1. In some embodiments, the anti-TNFα antibody binds to a protein comprising an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:1 or SEQ ID NO:2. In some embodiments, the anti-TNFα antibody binds to a protein comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:1 or SEQ ID NO:2. In some embodiments, the anti-TNFα antibody binds to a protein comprising an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:1 or SEQ ID NO:2.

[0120] Table 1: TNFα protein

[0121]

[0122] The anti-TNFα antibody can comprise a heavy chain comprising one or more amino acid sequences that are about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the sequences listed in Table 2.

[0123] The anti-TNFα antibody of the technology of the present invention comprises a heavy chain having at least one complementarity-determining region (CDR) sequence. At least one CDR sequence confers binding to a specific antigen (e.g., a TNFα peptide).

[0124] In some embodiments, at least one CDR sequence comprises an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the CDR sequences listed in Table 2 (e.g., SEQ ID NO: 3, 4, 5, 6, 7, or 8). In some embodiments, the anti-TNFα antibody comprises a heavy chain having a sequence comprising a CDR sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NOs: 3, 4, 5, 6, 7, and 8. The CDR sequences can be different from other CDR sequences (e.g., do not overlap with another CDR sequence), or at least a portion of the CDR sequence can overlap with a portion of another CDR sequence (e.g., two CDR sequences share at least a portion of the amino acid sequence).

[0125] In some embodiments, the anti-TNFα antibody comprises a heavy chain comprising a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 3.

[0126] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 3.

[0127] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 3.

[0128] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 4.

[0129] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 4.

[0130] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 4.

[0131] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO: 5.

[0132] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:5.

[0133] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:5.

[0134] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:6.

[0135] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:6.

[0136] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:6.

[0137] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:7.

[0138] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:7.

[0139] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:7.

[0140] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:8.

[0141] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:8.

[0142] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:8.

[0143] In some embodiments, the anti-TNFα antibody comprises a variable heavy (V H ) domain having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:9.

[0144] In some embodiments, the anti-TNFα antibody comprises a V H domain having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:9.

[0145] In some embodiments, the anti-TNFα antibody comprises a V H .

[0146] Table 2: Heavy chain anti-TNFα antibody sequences

[0147]

[0148] The anti-TNFα antibody can comprise a light chain that comprises one or more amino acid sequences that are about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the sequences listed in Table 3.

[0149] The anti-TNFα antibody of the present invention's technology comprises a light chain having at least one CDR sequence. The at least one CDR sequence confers binding to a specific antigen (e.g., TNFα peptide).

[0150] In some embodiments, the at least one CDR sequence comprises an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the CDR sequences listed in Table 2 (e.g., SEQ IDNO:10, 12, 13, 14, 15, or YA). In some embodiments, the anti-TNFα antibody comprises a light chain having a sequence that comprises CDR sequences that are about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:10, 12, 13, 14, 15, and YA. The CDR sequences can be different (e.g., do not overlap another CDR sequence) or can overlap (e.g., share at least a portion of the amino acid sequence).

[0151] In some embodiments, the anti-TNFα antibody comprises a light chain, and the light chain comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:10.

[0152] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:10.

[0153] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:10.

[0154] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to YA.

[0155] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:YA.

[0156] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:YA.

[0157] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:12.

[0158] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:12.

[0159] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:12.

[0160] In some embodiments, the anti-TNFα antibody comprises a CDR having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:13.

[0161] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:13.

[0162] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:13.

[0163] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:14.

[0164] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:14.

[0165] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:14.

[0166] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:15.

[0167] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:15.

[0168] In some embodiments, the anti-TNFα antibody comprises CDRs having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:15.

[0169] In some embodiments, the anti-TNFα antibody comprises a variable light (V L ) domain having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:16.

[0170] In some embodiments, the anti-TNFα antibody comprises a V domain having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:16. L domain.

[0171] In some embodiments, the anti-TNFα antibody comprises a V domain having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:16. L .

[0172] Table 3: Light chain anti-TNFα antibody sequences

[0173]

[0174] Anti-TNFα antibody function

[0175] The affinity of an anti-TNFα antibody for a TNFα protein or peptide is determined by the amino acid sequence on the heavy chain and / or light chain of each antibody. These amino acid sequences can include one or more CDRs. Binding of the anti-TNFα antibody to the TNFα protein or peptide can reduce the activity of TNFα. For example, the anti-TNFα antibody can reduce or prevent the consequences of TNFα signaling, including but not limited to nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling, mitogen-activated protein kinase (MAPK) signaling, pro-apoptotic signaling, cytokine and / or chemokine production (e.g., interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-12 (IL-12), interleukin-18 (IL-18), tumor necrosis factor beta (TNF-β), chemokine (C-X-C motif) ligand 1, 2, 5, 9, 10 and chemokine (C-C motif) ligand 2, 5 and 20) upregulation, cell adhesion molecule (e.g., intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin and P-selectin) and reactive oxygen species (ROS) production (e.g., superoxide anion, hydrogen peroxide and hydroxyl radical) upregulation.

[0176] In some embodiments, the binding of the anti-TNFα antibody to the TNFα protein or peptide can reduce the concentration of TNFα in the plasma or other tissues of a subject. For example, the anti-TNFα antibody can reduce membrane TNFα and / or cytoplasmic TNFα in resident cells, migratory cells, infiltrating cells, and / or intravascular cells (e.g., monocytes, epithelial cells, endothelial cells, glomerular cells, muscle cells, nerve cells, or myoepithelial cells) in the bladder, cervix, esophagus, large intestine, liver (Kupffer cells), lymph nodes, lung, pancreas, placenta (Hofbauer cells), parathyroid gland, salivary gland, skin, small intestine, spleen, stomach, thyroid gland, thymus, tonsils, and uterus.

[0177] Complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity:

[0178] The anti-TNFα antibody can mediate complement-dependent cytotoxicity (CDC) and / or antibody-dependent cell-mediated cytotoxicity (ADCC) of cells expressing mTNF. CDC or ADCC can be mediated by the binding of the anti-TNFα antibody to one or more of C1q, Fc receptors, FcγRI, FcγRIIb, or FcRn. CDC can determine the ability of the anti-TNFα antibody to induce cytotoxic effects on cells expressing TNFα in the presence of complement factors. ADCC can be mediated by NK cells as part of the humoral immune response. The ADCC of the anti-TNFα antibody can promote the lysis of cells expressing the TNFα protein or its peptide in the presence of effector cells of the immune system such as NK cells, muscle cells, macrophages, or eosinophils.

[0179] C1q Binding:As the first subcomponent of the C1 complex in the classical pathway of complement activation, C1q is considered to bind to sites on the CH2 domain of IgG. C1q contains the C1qA (accession number: NP_057075.1), C1qB (isoform 1 accession number: NP_000482.3; isoform 2 accession number: NP_001365085.1), and C1qC (isoform 1 accession number: NP_758957.2; isoform 2 accession number: NP_001334549.1) polypeptide chains. Together with two serine proteases, C1r (isoform 1 accession number: NP_001724.4; isoform 2 accession number: NP_001341275.1) and C1s (isoform 1 accession number: NP_001725.1; isoform 2 accession number: NP_001333779.1), C1q forms the complex C1, the first component of the complement. Activation of the complement cascade system generates C3 and C5 convertases. These enzymes produce the pro-inflammatory C3a and C5a anaphylatoxins and catalyze the formation of the pore-like membrane attack complex (C5-9), which can insert into the cell membrane by lysis or sub-lysis mechanisms, causing damage.

[0180] In some embodiments, the Fc portion of an anti-TNFα antibody or an antigen-binding fragment thereof that contains the Fc portion can bind to C1q or at least a portion of C1q (e.g., C1qA, C1qB, C1qC, or a peptide thereof). In some embodiments, the binding occurs at the CH2 domain of the anti-TNFα antibody. Binding to C1q or at least a portion of C1q can block C1q function and / or signaling. Non-limiting examples of C1q signaling include activation of the complement cascade system.

[0181] Fc Receptor Binding: CDC and / or ADCC can occur through the binding of an anti-TNFα antibody to Fc receptors (e.g., FcγRI, RII, and RIII), also known as CD64 (GenBank: AAI52384.1), CD32 (GenBank: AAI48274.1), and CD16 (NCBI: NP_001373379.1), respectively. In some embodiments, the Fc portion of an anti-TNFα antibody or an antigen-binding fragment thereof that contains the Fc portion can bind to an Fc receptor or at least a portion of an Fc receptor (e.g., FcγRI, RII, RIII, or a peptide thereof). Binding to the Fc receptor or at least said portion of the Fc receptor can trigger ADCC and cytotoxic cell activation and may result in the destruction of cells expressing TNFα.

[0182] FcγRI Binding:FcγRI (also known as CD64 (GenBank: AAI52384.1)) is a 72 kDa glycoprotein expressed on mononuclear phagocytes and can bind to IgG via its Fc-binding domain. FcγRI exhibits high affinity for IgG, Ka = 108 - 109 M-1, and can bind monomeric IgG. FcγRI released into biological fluids can act as an immunoglobulin-binding factor. FcγRI expression can be induced by interferon (IFN)-γ and can play a role in ADCC and the clearance of immune complexes. FcγRI binding can also be involved in the pathogenesis of immune complex- and autoantibody-mediated diseases. In some embodiments, the Fc portion of an anti-TNFα antibody or an antigen-binding fragment thereof comprising the Fc portion can bind to FcγRI or at least a portion of FcγRI. Binding to FcγRI can trigger ADCC and cytotoxic cell activation and may result in the destruction of cells expressing TNFα.

[0183] FcRn Binding: FcRn (Accession No.: AAF72596.1) consists of two subunits (an α subunit and β2-microglobulin). The α subunit is structurally related to major histocompatibility complex (MHC) class I proteins and can also associate with β2-microglobulin. The binding of IgG to pH-dependent FcRn protects antibodies from degradation via the endocytic salvage pathway, thereby prolonging the half-life of IgG in serum.

[0184] In some embodiments, an anti-TNFα antibody or an antigen-binding fragment thereof can bind to FcRn, at least a portion of FcRn (e.g., the α subunit, β2-microglobulin, or a peptide thereof). Binding to FcRn can trigger protection of the anti-TNFα antibody from degradation and / or prolong the half-life of the anti-TNFα antibody.

[0185] FcγRIIb Binding: The binding of an anti-TNFα antibody to FcγRIIb (isoform 1 Accession No.: NP_001381406.1) can modulate immune responses such as ADCC. FcγRIIb exhibits weak affinity for monomeric IgG, K a ≤107 M-1, and can only interact effectively with multimeric immune complexes. In some embodiments, an anti-TNFα antibody or an antigen-binding fragment thereof can bind to FcγRIIb or at least a portion of FcγRIIb. Binding to FcγRIIb can trigger ADCC and cytotoxic cell activation and may result in the destruction of cells expressing TNFα.

[0186] Post-translational modification

[0187] The anti-TNFα antibodies of the present invention's technology may comprise post-translational modifications (e.g., glycosylation, N-terminal glutaminyl cyclization, C-terminal lysine trimming, sialylation, methylation, acetylation, ubiquitination, sumoylation, phosphorylation, proteolytic cleavage), and such post-translational modifications may affect the characteristics of the antibody (e.g., immunogenicity, stability, affinity, dissociation rate). For example, the anti-TNFα antibody may be sialylated with one of a plurality of N-acetylneuraminic acid (Neu5Ac) molecules and / or may lack N-glycolylneuraminic acid (Neu5Gc). Neu5Ac is the predominant sialic acid present in human cells, while the major sialic acid form of other TNF inhibitors is Neu5Gc, which is only present in non-human cells and can be immunogenic.

[0188] In some embodiments, the anti-TNFα antibody is sialylated at Asn 297. In some embodiments, Asn 297 is sialylated with Neu5Ac.

[0189] In some embodiments, the anti-TNFα antibodies of the present invention's technology are glycosylated. In some embodiments, the anti-TNFα antibody is glycosylated at an asparagine residue (e.g., Asn 300) on at least one heavy chain. In some embodiments, the anti-TNFα antibody is glycosylated at an asparagine residue (e.g., Asn 300) on both heavy chains.

[0190] In some embodiments, the anti-TNFα antibodies of the present invention's technology are fucosylated.

[0191] In some embodiments, the anti-TNFα antibodies of the present invention's technology comprise one or more disulfide bonds (e.g., intra-chain or inter-chain). In some embodiments, relative to an antibody lacking disulfide bonds, one or more disulfide bonds enhance the stability and / or structural integrity of the antibody. In some embodiments, the anti-TNFα antibody comprises 16 disulfide bonds (12 intra-chain and 4 inter-chain).

[0192] In some embodiments, the anti-TNFα antibodies of the present invention's technology comprise a C-terminal lysine located on at least one heavy chain. In some embodiments, the anti-TNFα antibodies of the present invention's technology comprise a C-terminal lysine located on both heavy chains. In some embodiments, the C-terminal lysine is not trimmed. In some embodiments, the charge of the anti-TNFα antibody can be altered by performing C-terminal lysine trimming on one or both of the heavy chains.

[0193] In some embodiments, anti-TNFα antibodies can be produced using a biological system that affects the type of post-translational modification of the anti-TNFα antibody. For example, anti-TNFα antibodies can be produced using a Chinese hamster ovary (CHO) cell line rather than a cell line used to produce other TNFα inhibitors (e.g., SP2 cell line). Non-limiting examples of cell lines that can be used to produce anti-TNFα antibodies include CHO cells, NS0 cells, hybridoma cells, SP2 / 0 cells, human embryonic kidney (HEK) cells, PER.C6 cells, yeast cells, and myeloma cells.

[0194] Pharmacodynamics and Pharmacokinetics

[0195] The anti-TNFα antibodies of the technology of the present invention can include characteristics that alter pharmacodynamic parameters (e.g., binding, efficacy, and / or potency) and / or pharmacokinetic parameters (e.g., absorption, distribution, metabolism, or elimination) relative to a control (e.g., an anti-TNFα antibody that includes a sequence and / or a different post-translational modification state that is different from the sequence or post-translational modification state of the anti-TNFα antibody of the technology of the present invention). Characteristics of the anti-TNFα antibody (such as amino acid sequence or post-translational modification) may result in pharmacodynamic or pharmacokinetic differences relative to the control.

[0196] In some embodiments, the anti-TNFα antibodies of the technology of the present invention have an increased affinity for TNFα protein or its peptides relative to the affinity of the control. In some embodiments, the anti-TNFα antibodies of the technology of the present invention include an increased affinity for C1q protein, Fc receptor protein, FcγRI protein, FcγRIIb protein, or FcRn protein or their peptides.

[0197] Affinity can be evaluated by the dissociation constant (K D ) value. In some embodiments, the anti-TNFα antibodies of the technology of the present invention include a K D from TNFα protein or its peptides that is reduced relative to the K D of the control. In some embodiments, the anti-TNFα antibodies of the technology of the present invention include a K D from C1q protein, Fc receptor protein, FcγRI protein, FcγRIIb protein, or FcRn protein or their peptides that is reduced.

[0198] In some embodiments, the reduction in K D is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 99% reduction relative to the control.

[0199] In some embodiments, K DThe reduction is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 99% lower than the control.

[0200] In some embodiments, K D The reduction is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 99% lower than the control.

[0201] In some embodiments, the anti-TNFα antibody of the technology of the present invention has an increased half-life relative to the half-life of a control. The control may comprise an anti-TNFα antibody lacking post-translational modifications possessed by the anti-TNFα antibody of the technology of the present invention or other TNFα inhibitors.

[0202] In some embodiments, the increase in half-life is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900% or 1000% relative to the control.

[0203] In some embodiments, the increase in half-life is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900% or 1000% relative to the control.

[0204] In some embodiments, the increase in half-life is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900% or 1000% relative to the control.

[0205] In some embodiments, the anti-TNFα antibody of the technology of the present invention has a reduced clearance rate (CL) relative to the CL of a control. The control can include an anti-TNFα antibody lacking the post-translational modifications possessed by the anti-TNFα antibody of the technology of the present invention or other TNFα inhibitors. "Clearance rate" can refer to the elimination, absorption, and / or metabolism of the anti-TNFα antibody in the plasma of a subject. The clearance rate can be evaluated as the volume of plasma that clears the anti-TNFα antibody over time (e.g., milliliters per minute, liters per hour, or L / day) and / or can be normalized relative to the body weight of the subject (e.g., milliliters per minute per kilogram). A reduced clearance rate can also be indicated by an increase in the half-life or volume of distribution (V d ). In some embodiments, measuring the clearance rate includes measuring the terminal elimination rate constant (λ z ) or the intercompartmental clearance rate (Q).

[0206] In some embodiments, the reduced clearance rate includes measurements taken at about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0207] In some embodiments, the reduced clearance rate includes measurements taken at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0208] In some embodiments, the reduced clearance rate includes measurements taken at least about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0209] In some embodiments, the reduced clearance rate includes measurements taken during the administration of the anti-TNFα antibody.

[0210] In some embodiments, the reduced clearance rate includes measurements taken at the completion of the administration of the anti-TNFα antibody.

[0211] In some embodiments, the reduced clearance rate is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% lower than that of the control.

[0212] In some embodiments, the reduction in clearance is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% lower than the control.

[0213] In some embodiments, the reduction in clearance is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% lower than the control.

[0214] In some embodiments, the anti-TNFα antibody of the technology of the present invention has increased tissue, plasma, and / or serum concentrations relative to the tissue, plasma, and / or serum concentrations of a control. The control can be an anti-TNFα antibody lacking the post-translational modification possessed by the anti-TNFα antibody of the technology of the present invention or other TNFα inhibitors. "Concentration" can include measurements reflecting one or more of the absolute amount of the anti-TNFα antibody, the absorption of the anti-TNFα antibody, the metabolism of the anti-TNFα antibody, or the elimination of the anti-TNFα antibody.

[0215] The increase in tissue, plasma, and / or serum concentration can be an increase in the concentration of the anti-TNFα antibody at a given time point relative to the concentration of the control administered at the same dose and measured at the same time point. The concentration can be measured at an intermediate time point or a final time point and can be measured as the mean residence time (MRT), the average concentration (C 平均值 ), the trough concentration (C 谷 ), or the concentration at the end of the administration (e.g., infusion) time (C T ).

[0216] In some embodiments, the increase in concentration is reflected by an increase in the peak plasma concentration (C 最大值 ). The increase in C 最大值 can indicate increased absorption, decreased metabolism, or slower elimination of the anti-TNFα antibody relative to the control. In some embodiments, C 最大值 includes the dose-normalized C 最大值 (DNC 最大值 ).

[0217] In some embodiments, the increase in concentration is reflected by a decrease in the time (T 最大值 ) to reach C 最大值 . The decrease in T 最大值 can indicate increased absorption, decreased metabolism, or slower elimination of the anti-TNFα antibody relative to the control.

[0218] In some embodiments, the increased concentration is reflected by an increase in the area under the curve (AUC). The increase in AUC may mean an increase in the exposure of the anti-TNFα antibody relative to the control and / or may indicate increased absorption, decreased metabolism, or slower elimination of the anti-TNFα antibody. The AUC measurement result may include the area under the curve of the drug concentration (DNAUC) in non-compartmental analysis.

[0219] In some embodiments, the increased tissue, plasma, and / or serum concentration includes measurements taken at about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0220] In some embodiments, the increased tissue, plasma, and / or serum concentration includes measurements taken at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0221] In some embodiments, the increased tissue, plasma, and / or serum concentration includes measurements taken at least about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0222] In some embodiments, the increased tissue, plasma, and / or serum concentration includes measurements taken during administration of the anti-TNFα antibody.

[0223] In some embodiments, the increased tissue, plasma, and / or serum concentration includes measurements taken at the completion of administration of the anti-TNFα antibody.

[0224] In some embodiments, the increased tissue, plasma, and / or serum concentration is an increase of about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to the control.

[0225] In some embodiments, the increase in tissue, plasma, and / or serum concentration is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to the control.

[0226] In some embodiments, the increase in tissue, plasma, and / or serum concentration is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to the control.

[0227] In some embodiments, the distribution of the anti-TNFα antibody of the technology of the present invention is increased relative to the distribution of the control. The control can be an anti-TNFα antibody lacking the post-translational modification possessed by the anti-TNFα antibody of the technology of the present invention or other TNFα inhibitors.

[0228] The increase in distribution can be an increase in the distribution of the anti-TNFα antibody at a given time point relative to the distribution of the control administered at the same dose and measured at the same time point. The distribution can be measured at an intermediate time point or at the final time point.

[0229] In some embodiments, the measurement of the distribution includes measuring the distribution volume (V d or V dβ ) at the end stage, the central distribution volume (V), the peripheral distribution volume (V2), the apparent distribution volume (V z ), or the measurement of the distribution includes measuring the distribution volume (V ss ) at steady state. V d 、V dβ 、V z and / or V ss being high or increased can indicate a large distribution beyond the tissue, plasma, and / or serum compartments relative to the control.

[0230] In some embodiments, the increase in distribution includes measurements taken at about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0231] In some embodiments, the increased distribution comprises measurements taken at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0232] In some embodiments, the increased distribution comprises measurements taken at least about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the anti-TNFα antibody.

[0233] In some embodiments, the increased distribution comprises measurements taken during administration of the anti-TNFα antibody.

[0234] In some embodiments, the increased distribution comprises measurements taken at the completion of administration of the anti-TNFα antibody.

[0235] In some embodiments, the increased distribution is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control.

[0236] In some embodiments, the increased distribution is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control.

[0237] In some embodiments, the increased distribution is at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control.

[0238] Signal peptide

[0239] The anti-TNFα antibodies of the present invention's technology may comprise one or more signal peptides. The signal peptides may be present in the precursor and / or mature forms of the anti-TNFα antibodies and may direct the peptides of the anti-TNFα antibodies to specific cellular locations. Non-limiting examples of signal peptides include signal peptides that direct peptides to organelles (e.g., endoplasmic reticulum), signal peptides that facilitate peptide secretion, signal peptides that regulate protein sorting to subcellular components, signal peptides that facilitate peptide transmembrane translocation, and signal peptides that facilitate peptide insertion into membranes. In some embodiments, the signal peptides are cleaved.

[0240] In some embodiments, the signal peptide comprises SEQ ID NO:17 or SEQ ID NO:18 listed in Table 4.

[0241] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:17.

[0242] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:17.

[0243] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:17.

[0244] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:18.

[0245] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:18.

[0246] In some embodiments, the anti-TNFα antibody comprises a signal peptide having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:18.

[0247] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:19.

[0248] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:19.

[0249] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:19.

[0250] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and having an amino acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:20.

[0251] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and having an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:20.

[0252] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and having an amino acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:20.

[0253] Table 4: Signal Peptide

[0254]

[0255] Nucleic acid sequence

[0256] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:21.

[0257] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:21.

[0258] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ.ID NO:21.

[0259] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:22.

[0260] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:22.

[0261] In some embodiments, the anti-TNFα antibody comprises a signal peptide encoded by a nucleic acid sequence that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:22.

[0262] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:23.

[0263] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:23.

[0264] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:23.

[0265] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:24.

[0266] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is at least 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:24.

[0267] In some embodiments, the anti-TNFα antibody comprises a signal peptide located on the peptide chain and encoded by a nucleic acid sequence strand that is at least about 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ.ID NO:24.

[0268] Table 5: Nucleic Acid Sequences

[0269]

[0270]

[0271] Other anti-TNFα antibody characteristics

[0272] The anti-TNFα antibodies of the technology of the present invention may comprise isotypes that affect the functional properties and / or structural characteristics of the antibodies. In some embodiments, the anti-TNFα antibodies comprise isotypes selected from the group consisting of IgG, IgM, IgA, IgD, and IgE. The light chain may comprise isotype κ or isotype λ.

[0273] In some embodiments, the anti-TNFα antibodies of the technology of the present invention are chimeric antibodies. Non-limiting examples of chimeric antibodies include human-mouse antibodies, human-rat antibodies, human-hamster antibodies, human-rabbit antibodies, human-bovine antibodies, and human-goat antibodies. In some embodiments, the chimeric antibodies comprise humanized antibodies.

[0274] Compositions and Formulations

[0275] One or more anti-TNFα antibodies of the technology of the present invention may be present in a composition (e.g., a pharmaceutical composition). In some embodiments, the composition comprises a pharmaceutically acceptable carrier and / or excipient. The pharmaceutically acceptable carrier and / or excipient may comprise an aqueous or non-aqueous carrier that can facilitate the dissolution of the anti-TNFα antibody. Non-limiting examples of the pharmaceutically acceptable carrier and / or excipient include sucrose, polysorbate 80, monosodium phosphate, monohydrate, disodium phosphate, dihydrate. The composition may comprise a filler or diluent (e.g., lactose, mannitol, and microcrystalline cellulose), a disintegrant (e.g., croscarmellose sodium, crospovidone, starch), a buffer (e.g., phosphate buffer or acetate buffer), or a solvent (e.g., water, alcohol, glycerol).

[0276] In some embodiments, the composition does not contain a preservative.

[0277] In some embodiments, the composition is sterilized (e.g., by filtration).

[0278] In some embodiments, the composition comprises one or more anti-TNFα antibodies of the technology of the present invention in an amount of: about 0.1% w / v or w / w of the composition; at least about 1% w / v or w / w of the composition; about 10% w / v or w / w of the composition; about 20% w / v or w / w of the composition; about 30% w / v or w / w of the composition; about 40% w / v or w / w of the composition; about 50% w / v or w / w of the composition; about 60% w / v or w / w of the composition; about 70% w / v or w / w of the composition; about 80% w / v or w / w of the composition; about 90% w / v or w / w of the composition; about 95% w / v or w / w of the composition; or about 99% w / v or w / w of the composition.

[0279] In some embodiments, the composition comprises one or more anti-TNFα antibodies of the technology of the present invention in an amount of: at least 0.1% w / v or w / w of the composition; at least at least 1% w / v or w / w of the composition; at least 10% w / v or w / w of the composition; at least 20% w / v or w / w of the composition; at least 30% w / v or w / w of the composition; at least 40% w / v or w / w of the composition; at least 50% w / v or w / w of the composition; at least 60% w / v or w / w of the composition; at least 70% w / v or w / w of the composition; at least 80% w / v or w / w of the composition; at least 90% w / v or w / w of the composition; at least 95% w / v or w / w of the composition; or at least 99% w / v or w / w of the composition.

[0280] In some embodiments, the composition comprises one or more anti-TNFα antibodies of the technology of the present invention in an amount of: at least about 0.1% w / v or w / w of the composition; at least about 1% w / v or w / w of the composition; at least about 10% w / v or w / w of the composition; at least about 20% w / v or w / w of the composition; at least about 30% w / v or w / w of the composition; at least about 40% w / v or w / w of the composition; at least about 50% w / v or w / w of the composition; at least about 60% w / v or w / w of the composition; at least about 70% w / v or w / w of the composition; at least about 80% w / v or w / w of the composition; at least about 90% w / v or w / w of the composition; at least about 95% w / v or w / w of the composition; or at least about 99% w / v or w / w of the composition.

[0281] In some embodiments, the composition comprises a lyophilized drug product (lyophilized powder or cake of an anti-TNFα antibody). In some embodiments, the lyophilized drug product is reconstituted (e.g., using water or saline) prior to administration.

[0282] In some embodiments, the reconstituted composition provides a solution having a pH of about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8.0.

[0283] In some embodiments, the reconstituted composition provides a solution having a pH of at least 5.0, at least 5.1, at least 5.2, at least 5.3, at least 5.4, at least 5.5, at least 5.6, at least 5.7, at least 5.8, at least 5.9, at least 6.0, at least 6.1, at least 6.2, at least 6.3, at least 6.4, at least 6.5, at least 6.7, at least 6.8, at least 6.9, at least 7.0, at least 7.1, at least 7.2, at least 7.3, at least 7.4, at least 7.5, at least 7.6, at least 7.7, at least 7.8, at least 7.9, or at least 8.0.

[0284] In some embodiments, the reconstituted composition provides a solution having a pH of at least about 5.0, at least about 5.1, at least about 5.2, at least about 5.3, at least about 5.4, at least about 5.5, at least about 5.6, at least about 5.7, at least about 5.8, at least about 5.9, at least about 6.0, at least about 6.1, at least about 6.2, at least about 6.3, at least about 6.4, at least about 6.5, at least about 6.7, at least about 6.8, at least about 6.9, at least about 7.0, at least about 7.1, at least about 7.2, at least about 7.3, at least about 7.4, at least about 7.5, at least about 7.6, at least about 7.7, at least about 7.8, at least about 7.9, or at least about 8.0.

[0285] In some embodiments, the reconstituted composition provides a solution having an anti-TNFα antibody at a concentration of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 11 mg / mL, about 12 mg / mL, about 13 mg / mL, about 14 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / ML, about 30 mg / mL, or about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL or about 100 mg / mL.

[0286] In some embodiments, the reconstituted composition provides a solution having an anti-TNFα antibody at a concentration of at least 1 mg / mL, at least 2 mg / mL, at least 3 mg / mL, at least 4 mg / mL, at least 5 mg / mL, at least 6 mg / mL, at least 7 mg / mL, at least 8 mg / mL, at least 9 mg / mL, at least 10 mg / mL, at least 11 mg / mL, at least 12 mg / mL, at least 13 mg / mL, at least 14 mg / mL, at least 15 mg / mL, at least 20 mg / mL, at least 25 mg / ML, at least 30 mg / mL, or at least 40 mg / mL, at least 50 mg / mL, at least 60 mg / mL, at least 70 mg / mL, at least 80 mg / mL, at least 90 mg / mL or at least 100 mg / mL.

[0287] In some embodiments, the reconstituted composition provides a solution having an anti-TNFα antibody at a concentration of at least about 1 mg / mL, at least about 2 mg / mL, at least about 3 mg / mL, at least about 4 mg / mL, at least about 5 mg / mL, at least about 6 mg / mL, at least about 7 mg / mL, at least about 8 mg / mL, at least about 9 mg / mL, at least about 10 mg / mL, at least about 11 mg / mL, at least about 12 mg / mL, at least about 13 mg / mL, at least about 14 mg / mL, at least about 15 mg / mL, at least about 20 mg / mL, at least about 25 mg / ML, at least about 30 mg / mL, or at least about 40 mg / mL, at least about 50 mg / mL, at least about 60 mg / mL, at least about 70 mg / mL, at least about 80 mg / mL, at least about 90 mg / mL or at least about 100 mg / mL.

[0288] In some embodiments, the composition or compositions are diluted for administration. Non-limiting examples of diluents include sodium chloride, water (e.g., Sterile Water for Injection (SWFI)), dextrose solutions, bacteriostatic solutions, or Ringer's solution (e.g., a solution containing electrolytes, lactated Ringer's solution).

[0289] The composition can be formulated for intraperitoneal, intravenous, parenteral, subcutaneous, intramuscular, intraventricular, or oral administration. In some embodiments, the composition formulated for parenteral administration comprises the composition in the form of an aqueous or non-aqueous isotonic sterile injectable solution or suspension. As used herein, the term "parenteral" includes subcutaneous, intravenous, intraperitoneal, intramuscular, and intralesional or infusion techniques.

[0290] Related Methods

[0291] The present technology provides methods comprising administering one or more compositions of the present technology to a subject in need thereof. In some embodiments, the present technology comprises a method of treating or reducing sarcoidosis in a subject in need thereof relative to a control. In some embodiments, the present technology comprises a method of reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control. The control can refer to the subject at baseline, the subject at an intermediate time point, a second untreated subject, the second subject at an intermediate time point, or the second subject at baseline.

[0292] In some embodiments, the subject has one or more of the following: acute sarcoidosis, chronic sarcoidosis, progressive sarcoidosis, pulmonary sarcoidosis (e.g., with or without extrapulmonary manifestations), remitting sarcoidosis, refractory sarcoidosis, advanced sarcoidosis, ocular sarcoidosis, cutaneous sarcoidosis, or nervous system sarcoidosis. In some embodiments, the sarcoidosis comprises high responder or low responder sarcoidosis, wherein the names high responder sarcoidosis or low responder sarcoidosis can indicate the response to a therapeutic intervention in terms of in vitro granuloma formation or the response to beads coated with purified protein derivative (PPD).

[0293] In some embodiments, the subject has one or more granulomas. In some embodiments, the one or more granulomas comprise cutaneous granulomas, pulmonary granulomas, lymph node granulomas, ocular granulomas, hepatic granulomas, splenic granulomas, cardiac granulomas, or rheumatoid granulomas.

[0294] In some embodiments, the subject has a modified Medical Research Council (mMRC) dyspnea scale ≥ 1.

[0295] In some embodiments, the subject has a Scadding CXR grade determined by a chest X-ray based method. In some embodiments, the subject has a Scadding CXR grade selected from the group consisting of stage 0, stage II, stage III, or stage IV.

[0296] In some embodiments, the subject is at least 18 years of age. In some embodiments, the subject is between about 18 years and about 80 years of age. In some embodiments, the subject weighs at least 45 kg. In some embodiments, the subject weighs between about 45 kg and about 160 kg.

[0297] In some embodiments, the subject has sarcoidosis and / or one or more granulomas and one or more additional inflammatory conditions. In some embodiments, the inflammatory condition is an autoimmune disorder. Non-limiting examples of autoimmune disorders include rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), multiple sclerosis (MS), type 1 diabetes (T1D), psoriasis, celiac disease, Hashimoto's thyroiditis, Graves' disease, Crohn's disease, and myasthenia gravis (MG).

[0298] In some embodiments, the subject comprises anti-drug antibodies (ADA). The ADA can be produced by the subject's immune system in response to a therapeutic agent (e.g., a therapeutic agent comprising a TNFα inhibitor). The ADA can recognize and / or bind to the therapeutic agent, thereby potentially reducing the efficacy of the therapeutic agent (e.g., neutralizing the therapeutic agent and / or triggering an immune response against the therapeutic agent).

[0299] In some embodiments, the subject comprises neutralizing antibodies (nAB) that can inhibit the biological activity of a therapeutic agent (e.g., a therapeutic agent comprising a TNFα inhibitor). In the presence of the nAb, it can prevent the therapeutic agent from binding to its target or interfere with its function, thereby reducing its efficacy.

[0300] In some embodiments, the subject has received an anti-inflammatory drug. In some embodiments, the subject is receiving an anti-inflammatory drug. In some embodiments, the anti-inflammatory drug is selected from the group consisting of corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, ezogabine, rituximab, adalimumab, golimumab, natalizumab, and infliximab.

[0301] In some embodiments, the corticosteroid is selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone, and triamcinolone. In some embodiments, the subject has received or is receiving from about 7.5 mg / day to about 25 mg / day of corticosteroid.

[0302] In some embodiments, the subject has received or is receiving a therapeutic agent selected from the group consisting of: short-acting β-agonist (SABA) (e.g., albuterol / salbutamol), short-acting muscarinic antagonist (SAMA) (e.g., ipratropium bromide), long-acting β-agonist (LABA) (e.g., formoterol or salmeterol), ultra-long-acting β-agonist (Ultra-LABA) (e.g., indacaterol, vilanterol, or olodaterol), and long-acting muscarinic antagonist (LAMA) (e.g., tiotropium, umeclidinium, aclidinium, or glycopyrronium).

[0303] Dose / Period

[0304] In some embodiments, the composition is administered to the subject at a first dose. The first dose can comprise administering the composition to the subject in an amount of about 0.5 mg / kg, about 1.0 mg / kg, about 1.5 mg / kg, about 2.0 mg / kg, about 2.5 mg / kg, about 3.0 mg / kg, about 3.5 mg / kg, about 4.0 mg / kg, about 4.5 mg / kg, about 5.0 mg / kg, about 5.5 mg / kg, or about 6.0 mg / kg.

[0305] In some embodiments, the composition is administered to the subject at a first dose in an amount of at least 0.5 mg / kg, at least 1.0 mg / kg, at least 1.5 mg / kg, at least 2.0 mg / kg, at least 2.5 mg / kg, at least 3.0 mg / kg, at least 3.5 mg / kg, at least 4.0 mg / kg, at least 4.5 mg / kg, at least 5.0 mg / kg, at least 5.5 mg / kg, or at least 6.0 mg / kg.

[0306] In some embodiments, the composition is administered to a subject in a first dose of at least about 0.5 mg / kg, at least about 1.0 mg / kg, at least about 1.5 mg / kg, at least about 2.0 mg / kg, at least about 2.5 mg / kg, at least about 3.0 mg / kg, at least about 3.5 mg / kg, at least about 4.0 mg / kg, at least about 4.5 mg / kg, at least about 5.0 mg / kg, at least about 5.5 mg / kg, or at least about 6.0 mg / kg.

[0307] In some embodiments, the first dose of the composition is administered about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, or about once every six weeks for a first period of time.

[0308] In some embodiments, the first dose of the composition is administered at least once a week, at least once every two weeks, at least once every three weeks, at least once every four weeks, at least once every five weeks, or at least once every six weeks for a first period of time.

[0309] In some embodiments, the first dose of the composition is administered at least about once a week, at least about once every two weeks, at least about once every three weeks, at least about once every four weeks, at least about once every five weeks, or at least about once every six weeks for a first period of time.

[0310] In some embodiments, the first period of time is about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 18 weeks, about 20 weeks, about 22 weeks, or about 24 weeks.

[0311] In some embodiments, the first period of time is at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 18 weeks, at least 20 weeks, at least 22 weeks, or at least 24 weeks.

[0312] In some embodiments, the first period of time is at least about 6 weeks, at least about 8 weeks, at least about 10 weeks, at least about 12 weeks, at least about 14 weeks, at least about 15 weeks, at least about 16 weeks, at least about 18 weeks, at least about 20 weeks, at least about 22 weeks, or at least about 24 weeks.

[0313] In some embodiments, a cumulative dose of the composition of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg is administered to a subject during a first time period.

[0314] In some embodiments, a composition having a cumulative dose of at least 2 mg / kg, at least 3 mg / kg, at least 4 mg / kg, at least 5 mg / kg, at least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 10 mg / kg, at least 11 mg / kg, at least 12 mg / kg, at least 13 mg / kg, at least 14 mg / kg, at least 15 mg / kg, at least 16 mg / kg, at least 17 mg / kg, at least 18 mg / kg, at least 19 mg / kg, or at least 20 mg / kg is administered to a subject during a first time period.

[0315] In some embodiments, a composition having a cumulative dose of at least about 2 mg / kg, at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, at least about 18 mg / kg, at least about 19 mg / kg, or at least about 20 mg / kg is administered to a subject during a first time period.

[0316] In some embodiments, the composition is administered to the subject in a second dose. In some embodiments, the second dose comprises administering the composition to the subject in an amount of about 0.5 mg / kg, about 1.0 mg / kg, about 1.5 mg / kg, about 2.0 mg / kg, about 2.5 mg / kg, about 3.0 mg / kg, about 3.5 mg / kg, about 4.0 mg / kg, about 4.5 mg / kg, about 5.0 mg / kg, about 5.5 mg / kg, or about 6.0 mg / kg.

[0317] In some embodiments, the composition is administered to a subject at a second dose in an amount of at least 0.5 mg / kg, at least 1.0 mg / kg, at least 1.5 mg / kg, at least 2.0 mg / kg, at least 2.5 mg / kg, at least 3.0 mg / kg, at least 3.5 mg / kg, at least 4.0 mg / kg, at least 4.5 mg / kg, at least 5.0 mg / kg, at least 5.5 mg / kg, or at least 6.0 mg / kg.

[0318] In some embodiments, the composition is administered to a subject at a second dose in an amount of at least about 0.5 mg / kg, at least about 1.0 mg / kg, at least about 1.5 mg / kg, at least about 2.0 mg / kg, at least about 2.5 mg / kg, at least about 3.0 mg / kg, at least about 3.5 mg / kg, at least about 4.0 mg / kg, at least about 4.5 mg / kg, at least about 5.0 mg / kg, at least about 5.5 mg / kg, or at least about 6.0 mg / kg.

[0319] In some embodiments, the second dose of the composition is administered about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once every 5 weeks, or about once every 6 weeks for a second period of time.

[0320] In some embodiments, the second dose of the composition is administered at least once a week, at least once every two weeks, at least once every three weeks, at least once every four weeks, at least once every 5 weeks, or at least once every 6 weeks for a second period of time.

[0321] In some embodiments, the second dose of the composition is administered at least about once a week, at least about once every two weeks, at least about once every three weeks, at least about once every four weeks, at least about once every 5 weeks, or at least about once every 6 weeks for a second period of time.

[0322] In some embodiments, the second period of time is about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 18 weeks, about 20 weeks, about 22 weeks, or about 24 weeks.

[0323] In some embodiments, the second period of time is at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 18 weeks, at least 20 weeks, at least 22 weeks, or at least 24 weeks.

[0324] In some embodiments, the second period of time is at least about 6 weeks, at least about 8 weeks, at least about 10 weeks, at least about 12 weeks, at least about 14 weeks, at least about 15 weeks, at least about 16 weeks, at least about 18 weeks, at least about 20 weeks, at least about 22 weeks, or at least about 24 weeks.

[0325] In some embodiments, the second dose is about twice the first dose.

[0326] In some embodiments, the second dose is at least twice the first dose.

[0327] In some embodiments, the second dose is at least about twice the first dose.

[0328] In some embodiments, the second dose is substantially the same as the first dose.

[0329] In some embodiments, the administration frequency of the second dose includes the same administration frequency as that of the first dose.

[0330] In some embodiments, the administration frequency of the second dose consists of the same administration frequency as that of the first dose.

[0331] In some embodiments, the administration frequency of the second dose is about twice the administration frequency of the first dose.

[0332] In some embodiments, each of the first dose and the second dose is administered about once every four weeks.

[0333] In some embodiments, the first dose is administered about once every four weeks, and the second dose is administered about once every two weeks.

[0334] In some embodiments, a composition having a cumulative dose of about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg is administered to a subject during a second time period.

[0335] In some embodiments, a composition having a cumulative dose of at least 5 mg / kg, at least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 10 mg / kg, at least 11 mg / kg, at least 12 mg / kg, at least 13 mg / kg, at least 14 mg / kg, at least 15 mg / kg, at least 16 mg / kg, at least 17 mg / kg, at least 18 mg / kg, at least 19 mg / kg, or at least 20 mg / kg is administered to a subject during a second time period.

[0336] In some embodiments, a composition is administered to a subject during a second time period at a cumulative dose of at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, at least about 18 mg / kg, at least about 19 mg / kg, or at least about 20 mg / kg.

[0337] In some embodiments, the composition is administered to the subject at a third dose. In some embodiments, the third dose comprises administering the composition to the subject in an amount of about 0.5 mg / kg, about 1.0 mg / kg, about 1.5 mg / kg, about 2.0 mg / kg, about 2.5 mg / kg, about 3.0 mg / kg, about 3.5 mg / kg, about 4.0 mg / kg, about 4.5 mg / kg, about 5.0 mg / kg, about 5.5 mg / kg, or about 6.0 mg / kg.

[0338] In some embodiments, the composition is administered to the subject at a third dose in an amount of at least 0.5 mg / kg, at least 1.0 mg / kg, at least 1.5 mg / kg, at least 2.0 mg / kg, at least 2.5 mg / kg, at least 3.0 mg / kg, at least 3.5 mg / kg, at least 4.0 mg / kg, at least 4.5 mg / kg, at least 5.0 mg / kg, at least 5.5 mg / kg, or at least 6.0 mg / kg.

[0339] In some embodiments, the composition is administered to the subject at a third dose in an amount of at least about 0.5 mg / kg, at least about 1.0 mg / kg, at least about 1.5 mg / kg, at least about 2.0 mg / kg, at least about 2.5 mg / kg, at least about 3.0 mg / kg, at least about 3.5 mg / kg, at least about 4.0 mg / kg, at least about 4.5 mg / kg, at least about 5.0 mg / kg, at least about 5.5 mg / kg, or at least about 6.0 mg / kg.

[0340] In some embodiments, the third dose of the composition is administered about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once every 5 weeks, or about once every 6 weeks for a third time period.

[0341] In some embodiments, the third dose of the composition is administered at least once a week, at least once every two weeks, at least once every three weeks, at least once every four weeks, at least once every 5 weeks, or at least once every 6 weeks for a third time period.

[0342] In some embodiments, the third dosing regimen composition is administered at least about once a week, at least about once every two weeks, at least about once every three weeks, at least about once every four weeks, at least about once every five weeks, or at least about once every six weeks for a third time period.

[0343] In some embodiments, the third time period is about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 18 weeks, about 20 weeks, about 22 weeks, or about 24 weeks.

[0344] In some embodiments, the third time period is at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 18 weeks, at least 20 weeks, at least 22 weeks, or at least 24 weeks.

[0345] In some embodiments, the third time period is at least about 6 weeks, at least about 8 weeks, at least about 10 weeks, at least about 12 weeks, at least about 14 weeks, at least about 15 weeks, at least about 16 weeks, at least about 18 weeks, at least about 20 weeks, at least about 22 weeks, or at least about 24 weeks.

[0346] In some embodiments, a composition having a cumulative dose of about 20 mg / kg, about 22 mg / kg, about 24 mg / kg, about 25 mg / kg, about 26 mg / kg, about 28 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 44 mg / kg, about 46 mg / kg, about 48 mg / kg, about 50 mg / kg, about 52 mg / kg, about 54 mg / kg, about 56 mg / kg, about 58 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, or about 75 mg / kg is administered to a subject over all time periods.

[0347] In some embodiments, a composition having a cumulative dose of at least 20 mg / kg, at least 22 mg / kg, at least 24 mg / kg, at least 25 mg / kg, at least 26 mg / kg, at least 28 mg / kg, at least 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 44 mg / kg, at least 46 mg / kg, at least 48 mg / kg, at least 50 mg / kg, at least 52 mg / kg, at least 54 mg / kg, at least 56 mg / kg, at least 58 mg / kg, at least 60 mg / kg, at least 65 mg / kg, at least 70 mg / kg, or at least 75 mg / kg is administered to a subject over all time periods.

[0348] In some embodiments, a composition is administered to a subject at a cumulative dose of at least about 20 mg / kg, at least about 22 mg / kg, at least about 24 mg / kg, at least about 25 mg / kg, at least about 26 mg / kg, at least about 28 mg / kg, at least about 30 mg / kg, at least about 35 mg / kg, at least about 40 mg / kg, at least about 44 mg / kg, at least about 46 mg / kg, at least about 48 mg / kg, at least about 50 mg / kg, at least about 52 mg / kg, at least about 54 mg / kg, at least about 56 mg / kg, at least about 58 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, or at least about 75 mg / kg across all time periods.

[0349] In some embodiments, the composition is administered using an infusion (e.g., intravenous infusion). In some embodiments, the infusion is administered over a time period of about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, or about 5 hours.

[0350] In some embodiments, the infusion is administered over a time period of at least 30 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, or at least 5 hours.

[0351] In some embodiments, the infusion is administered over a time period of at least about 30 minutes, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, or at least about 5 hours.

[0352] Results of administering and using the composition

[0353] In some embodiments, relative to a control, a subject administered a composition of the technology of the present invention experiences a decrease in the level of one or more transcripts selected from the group consisting of angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL6), interleukin 2 receptor (IL2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL1B), interleukin 12 (IL12), interleukin 18 (IL18), interferon γ (IFNG), interleukin 8 (IL8), monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), chemokine ligand 2 (CXCL2), chemokine ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JunB), mitogen-activated protein kinase (MKP1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL32), intercellular adhesion molecule 1 (ICAM-1), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), and vascular cell adhesion molecule 1 (VCAM-1).

[0354] In some embodiments, relative to a control, a subject administered a composition of the technology of the present invention experiences a decrease in the level of one or more proteins selected from the group consisting of: angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL-6), interleukin 2 receptor (IL-2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL-1B), interferon γ (IFN-γ), interleukin 8 (IL-8), interleukin 12 (IL-12), interleukin 18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), chemokine (C-X-C motif) ligand 1 (CXCL1), chemokine (C-X-C motif) ligand 2 (CXCL2), chemokine (C-X-C motif) ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JUNB), mitogen-activated protein kinase phosphatase 1 (MKP-1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL-32), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1).

[0355] In some embodiments, relative to a control, a subject administered a composition of the technology of the present invention experiences an increase in one or more of the following: forced vital capacity (FVC) measurement, forced expiratory volume in 1 second (FEV1) measurement, Leicester Cough Questionnaire (LCQ) score, Short Form 36 Health Survey (SF-36) score, King's Sarcoidosis Questionnaire (KSQ) score (e.g., KSQ general health status module or KSQ lung score), steroid toxicity scale (STS) score, or 6-minute walk test (6MWT) distance.

[0356] In some embodiments, relative to a control, a subject administered a composition of the technology of the present invention experiences a decrease in one or more of the following: patient global assessment (PGA) score, fatigue assessment scale (FAS) score, modified Medical Research Council (mMRC) dyspnea scale score, St. George's Respiratory Questionnaire (SGRQ) score, or Borg CR10 dyspnea score.

[0357] In some embodiments, subjects administered the compositions of the technology of the present invention experience a decrease in the stage of Scadding CXR relative to a control.

[0358] In some embodiments, subjects administered the compositions of the technology of the present invention experience an improvement in quality of life relative to a control. In some embodiments, quality of life is assessed by one or more of a KSQ score, an LCQ score, a PGA score, a FAS score, a mMRC dyspnea scale score, an SF-36 score, or a steroid toxicity questionnaire (STQ) score.

[0359] In some embodiments, subjects administered the compositions of the technology of the present invention experience an improvement in one or more limitations selected from the group consisting of: limitations on physical activity due to health problems, limitations on social activity due to physical or emotional problems, limitations on usual role activities due to physical health problems, and limitations on usual role activities due to emotional problems.

[0360] In some embodiments, subjects administered the compositions of the technology of the present invention experience a reduction in body pain relative to a control.

[0361] In some embodiments, subjects administered the compositions of the technology of the present invention experience an improvement in mental health relative to a control.

[0362] In some embodiments, subjects administered the compositions of the technology of the present invention experience an increase in vitality (e.g., an increase in energy and / or a reduction in fatigue) relative to a control.

[0363] In some embodiments, subjects administered the compositions of the technology of the present invention experience an improvement in the perception of general health relative to a control.

[0364] In some embodiments, subjects administered the compositions of the technology of the present invention experience an increase in lifespan relative to a control.

[0365] In some embodiments, subjects administered the compositions of the technology of the present invention experience a decrease in the need for anti-inflammatory drugs (e.g., the frequency, dose, or duration of administration) relative to a control. In some embodiments, the anti-inflammatory drugs are selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, ezogabine, rituximab, adalimumab, golimumab, namilumab, and infliximab. In some embodiments, the corticosteroids are selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone, and triamcinolone.

[0366] In some embodiments, subjects administered a composition of the technology of the present invention experience a reduction in granuloma formation or a decrease in granuloma size relative to a control. In some embodiments, the granuloma is a cutaneous granuloma, a pulmonary granuloma, a lymph node granuloma, an ocular granuloma, a hepatic granuloma, a splenic granuloma, a cardiac granuloma, or a rheumatoid granuloma.

[0367] In some embodiments, subjects administered a composition of the technology of the present invention experience a decrease in the levels of calcitriol, neopterin, soluble IL-2R (sIL-2R), or lysozyme in the subject relative to a control.

[0368] In some embodiments, subjects administered a composition of the technology of the present invention experience a decrease in immune cell energy relative to a control.

[0369] In some embodiments, subjects administered a dose of one or more compositions of the technology of the present invention exhibit one or more of the following:

[0370] (a) a reduction in ACE2 transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0371] (b) a reduction in IL6 transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0372] (c) a reduction in IL2R transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0373] (d) a reduction in TNFα transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0374] (e) a reduction in CRP transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0375] (f) a reduction in IL1B transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0376] (g) a reduction in IL12 transcript level of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;

[0377] (h) The level of IL18 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0378] (i) The level of IFNG transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0379] (j) The level of IL8 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0380] (k) The level of MCP-1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0381] (l) The level of CXCL1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0382] (m) The level of CXCL2 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0383] (o) The level of CXCL3 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0384] (p) The level of EGR1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0385] (q) The level of JunB transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0386] (r) The level of MKP1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0387] (s) The level of TNFAIP1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0388] (t) The level of TNFAIP2 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0389] (u) The level of TNFAIP3 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0390] (v) The level of SDC4 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0391] (w) The level of SOD2 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0392] (x) The level of COX2 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0393] (y) The level of IL32 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0394] (z) The level of ICAM-1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0395] (aa) The level of KL-6 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0396] (bb) The level of CXCL10 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0397] (cc) The level of CXCL9 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0398] (dd) The level of CHIT1 transcript is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0399] (ee) The SCGB1A1 transcript level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0400] (ff) The SAA1 transcript level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0401] (gg) The VCAM-1 transcript level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0402] (hh) The ACE2 protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0403] (ii) The IL-6 protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0404] (jj) The IL-2R protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0405] (kk) The TNFα protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0406] (ll) The CRP protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0407] (mm) The IL-1B protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0408] (nn) The MCP-1 protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0409] (oo) The CXCL1 protein level was reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0410] (pp) The CXCL2 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0411] (qq) The CXCL3 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0412] (rr) The EGR1 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0413] (ss) The JUNB protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0414] (tt) The MKP-1 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0415] (uu) The TNFAIP1 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0416] (vv) The TNFAIP2 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0417] (ww) The TNFAIP3 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0418] (xx) The SDC4 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0419] (yy) The SOD2 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0420] (zz) The COX2 protein level was reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0421] (aaa) The level of IL-32 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0422] (bbb) The level of KL-6 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0423] (ccc) The level of CXCL10 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0424] (ddd) The level of CXCL9 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0425] (eee) The level of CHIT1 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0426] (fff) The level of SCGB1A1 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0427] (ggg) The level of SAA1 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0428] (hhh) The level of ICAM-1 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0429] (iii) The level of VCAM-1 protein is reduced by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0430] (jjj) The FVC measurement result is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0431] (kkk) The FEV1 measurement result is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0432] (lll) The LCQ score increased by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, or 18 points relative to the control;

[0433] (mmm) The SF-36 score increased by at least 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points, or 100 points relative to the control;

[0434] (nnn) The KSQ score increased by at least 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points, or 100 points relative to the control;

[0435] (ooo) The STS score increased by at least 1 point, 2 points, 3 points, 4 points, 5 points or 6 points relative to the control;

[0436] (ppp) The 6MWT distance increased by at least 10 m, 20 m, 30 m, 40 m, 50 m, 60 m, 70 m, 80 m, 90 m, 100 m, 125 m, 150 m, 175 m, 200 m, 250 m, 300 m, 350 m, 400 m, 450 m or 500 m relative to the control;

[0437] (qqq) The PGA score decreased by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points or 10 points relative to the control;

[0438] (rrr) The FAS score decreased by at least about 1 point, 2 points, 3 points or 4 points relative to the control;

[0439] (sss) The mMRC scale score decreased by at least about 1 level, 2 levels, 3 levels or 4 levels relative to the control;

[0440] (ttt) The SGRQ score decreased by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points or 100 points relative to the control;

[0441] (uuu) The Borg CR10 dyspnea score decreased by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points or 10 points relative to the control;

[0442] (vvv) Granuloma formation was reduced by about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0443] (www) Granuloma size was decreased by about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0444] (xxx) The calcitriol level was decreased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0445] (yyy) The neopterin level was decreased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0446] (zzz) The lysozyme level was decreased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control;

[0447] (aaaa) The sIL-2R level was decreased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control; and

[0448] (bbbb) The Scadding CXR stage was increased by at least 1, 2, 3 or 4 stages relative to the control.

[0449] Example 1: Evaluation of the Major Pharmacodynamics of XTMAB - 16

[0450] Inhibition of TNFα-mediated cytotoxicity

[0451] XTMAB-16 is an immunosuppressive and immunomodulatory agent due to its binding to TNFα. The potency of XTMAB-16 was tested by measuring the inhibition of TNFα-induced cytotoxicity in a melanoma cell line expressing the tumor necrosis factor receptor (TNFR). Cells from the melanoma cell line A375 (ATCC CRL-1619) were incubated with TNFα, actinomycin-D, and XTMAB-16 at concentrations from 1 ng / mL to 1000 ng / mL for 24 hours. Cell density was then determined by luminescence. The potency of XTMAB-16 was compared to the potency of another TNFα blocker (infliximab), as Figure 1 shown.

[0452] The potencies of experimental batches containing XTMAB-16 and infliximab are shown in Table 6. The potency of the XTMAB-16 batches was 90% to 103% relative to infliximab.

[0453] Table 6: Relative Potency of XTMAB-16

[0454]

[0455]

[0456] Abbreviations: DP = drug product; DS = drug substance; RS = reference standard

[0457] Transmembrane TNFα (mTNFα) Binding

[0458] A cell-based enzyme-linked immunosorbent assay (ELISA) was used to test the binding of XTMAB-16 to mTNFα. The assay used a cell line (T3-33) expressing mTNFα developed from SP2 / 0 cells (mouse myeloma cells). T3-33 cells were incubated with XTMAB16 or infliximab at 5 ng / mL to 3000 ng / mL in 3% bovine serum albumin (BSA). Bound antibody was detected using horseradish peroxidase (HRP) conjugated to mouse anti-human IgG1 and the chromogen 3,3',5,5'-tetramethylbenzidine (TMB). Binding of XTMAB-16 to TNFα expressed on the cell membrane was then detected at 450 nm ( Figure 2 ). XTMAB-16 bound to mTNFα with 91% to 95%, as shown in Table 7.

[0459] Table 7: mTNFα Binding, Cell-based ELISA

[0460]

[0461] Abbreviations: ELISA = enzyme-linked immunosorbent assay; mTNFα = transmembrane tumor necrosis factor α; No. = number. Note: Results from duplicate measurements are presented as mean values.

[0462] a Binding relative to infliximab

[0463] TNFα binding kinetics

[0464] A biosensor coated with the antibody's anti-human IgG fragment crystallizable region (Fc) was used to capture the antibody and immobilize XTMAB-16, which was incubated with different levels of recombinant TNFα to determine the binding kinetics between TNFα and the antigen-binding fragment antibody region (Fab) domain of XTMAB-16.

[0465] Representative kinetic curves of TNFα binding to XTMAB-16 are presented in Figure 3 . Kinetic parameters for different batches of XTMAB16 are presented in Table 8.

[0466] Determine the binding kinetics of TNFα to XTMAB16 at an affinity constant (K -11 ) of 5.68 to 6.64×10 D M.

[0467] Table 8: TNFα binding kinetics

[0468]

[0469] Abbreviations: k D = affinity constant; k 解离 = dissociation constant; k 缔合 = association constant; No. = number;

[0470] TNFα = tumor necrosis factor α.

[0471]

[0472] Abbreviations: k D = affinity constant; k 解离 = dissociation constant; k 缔合 = association constant; No. = number;

[0473] TNFα = tumor necrosis factor α.

[0474] Cross-species binding study

[0475] The binding of XTMAB16 to human TNFα was confirmed by indirect sandwich ELISA and direct ELISA. The binding of XTMAB16 to human TNFα was higher than 1,000 pg / mL at the calculated half maximal effective concentration (EC50) of 60,675 pg / mL as observed by indirect sandwich ELISA. As determined by the direct ELISA method, the binding of XTMAB16 to human TNFα was higher than 10,000 pg / mL at the calculated EC50 of 5.78×10 5 pg / mL. The binding of human capture antibody to human TNFα was higher than 10,000 pg / mL at the calculated EC50 of 7.37×10 6 pg / mL. XTMAB-16 did not bind to TNFα from mouse, rat, dog, or rhesus or cynomolgus monkeys. These data indicate that none of the traditional non-clinical species are considered relevant for the assessment of the safety of XTMAB16.

[0476] Tissue cross-reactivity study

[0477] In a Good Laboratory Practice (GLP) study, the cross-reactivity of XTMAB-16 in cryosections from a complete panel consisting of normal human tissues and human sarcoidosis tissues was investigated. The tissue panel used as the test system included the "Suggested list of human tissues for immunohistochemical or cytochemical studies of cross-reactivity of monoclonal antibodies" in Annex I of the European Medicines Agency (EMA) document "Guideline on Development, Production, Characterisation, and Specifications for Monoclonal Antibodies and Related Products" adopted by the Committee for Medicinal Products for Human Use (CHMP). All tissues on "human tissues to be used for immunohistochemical or cytochemical investigations of crossreactivity of monoclonal antibodies)" and all tissues recommended in the document "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use" by the Center for Biologics Evaluation and Research (CBER) of the US Food and Drug Administration (FDA). Tonsils, human lungs and lymph nodes of patients with sarcoidosis and small intestines of patients with Crohn's disease were also included in the study.

[0478] Samples from each of the listed tissues from 1 to 3 individual donors were evaluated. Fresh unfixed tissue samples were placed in molds filled with Tissue- OCT compound and frozen at -85 °C to -70 °C until sectioning. Sections were cut at approximately 5 μm and fixed in acetone for 10 minutes at room temperature. Human blood smears were stored at -85 °C to -70 °C until fixation and staining. XTMAB16 (primary antibody) was incubated overnight with biotinylated donkey anti-human IgG secondary labeled antibody. XTMAB16 and control product hIgG1 were tested at concentrations of 5 μg / mL and 20 μg / mL. Immunopathology was evaluated by light microscopy.

[0479] XTMAB16 produced weak to strong membrane and cytoplasmic staining of rare or occasional positive control T3-33 (expressing TNFα) cells at both staining concentrations. The antibody did not react specifically with negative control frozen sections. The control product did not react specifically with positive or negative control materials, and the control slides were unstained. The specific reaction of XTMAB16 with positive control materials, the lack of specific reactivity with negative control materials, and the lack of reactivity with the control product in all staining runs indicated that the assay was sensitive, specific, and reproducible.

[0480] The results of these studies are shown in Table 9.

[0481] Table 9: Binding of XTMAB-16 to normal human tissues.

[0482]

[0483]

[0484] The staining of monocytes, glial cells, and spindle cells observed with XTMAB-16 was consistent with TNFα expression in macrophages, T lymphocytes, NK cells, microglia, and fibroblasts. Additionally, the staining observed with XTMAB-16 on endometrial epithelial cells and placental decidual cells was consistent with the reported expression of TNFα. The membrane and cytoplasmic binding of XTMAB-16 in epithelial cells of the liver and myoepithelial cells of the breast was not described in the literature. Although the subcellular localization of the binding of XTMAB-16 to the neuropil could not be definitively determined using light microscopy, the binding was considered to have limited in vivo significance due to the restricted access of the mAb across the blood-brain barrier.

[0485] The remaining staining performed with XTMAB-16 was cytoplasmic in nature, and according to ICH S6(R1) and other literature, the binding of the mAb to cytoplasmic sites is generally considered to have little toxicological significance since monoclonal therapeutic agents generally cannot enter the cytoplasmic compartment in vivo. In addition, it should be noted that the binding of XTMAB-16 to most non-monocytes in this study was of low affinity (i.e., binding was only observed at higher concentrations of XTMAB-16 in most tissues).

[0486] In tissues obtained from subjects, monocytes in these tissues were selected as secondary controls since TNFα expression was reported in monocytes in human tonsils, small intestine with Crohn's disease, lungs with sarcoidosis, and lymph nodes with sarcoidosis. The purpose of the secondary control tissues was to exhibit XTMAB-16 specific binding in the presence of relevant cell types. XTMAB-16 weakly to strongly stained the membrane and cytoplasm of rare to occasional monocytes in human tonsils at higher staining concentrations and weak to strong and rare staining was noted at lower concentrations. XTMAB-16 also weakly to strongly stained the membrane and cytoplasm of rare to occasional monocytes in the small intestine of humans with Crohn's disease, lungs with sarcoidosis, and lymph nodes with sarcoidosis at higher staining concentrations and medium to rare staining at lower concentrations. No staining was performed on the secondary control tissues on the assay control slides.

[0487] This immunohistochemical study provides support related to the mechanism of XTMAB-16 as a TNFα blocker. TNFα is a pleiotropic cytokine produced by macrophages, monocytes, neutrophils, T cells, NK cells, and fibroblasts. Alveolar macrophage-derived TNFα is involved in the induction and maintenance of granulomas in sarcoidosis, and high levels of TNFα (and high levels of TNFα released from alveolar macrophages) appear to be associated with disease progression. Similar inflammatory mechanisms are believed to underlie sarcoidosis involvement in other organs such as the skin. In this study, cross-reactivity of human tissues with XTMAB-16 was expected in both normal and diseased samples. Thus, these immunohistochemical results support the use of XTMAB-16 in subjects with sarcoidosis and are not expected to cause toxicity in tissues not affected by sarcoidosis.

[0488] In Vivo Pharmacology

[0489] In a 6-week repeated-dose toxicity study, the in vivo pharmacology of XTMAB-16 was evaluated in transgenic TC (TgTC) mice (a rheumatoid arthritis model). XTMAB-16 was administered to TgTC mice (10 per sex / dose) at doses of 0 mg / kg / dose, 20 mg / kg / dose, and 40 mg / kg / dose every two weeks on days 1, 15, and 29, along with a 2-week recovery period.

[0490] The inhibitory effect of XTMAB-16 as a TNFα blocker and the potential pharmacologic benefits of XTMAB-16 were evaluated by comparing arthritis scores and histopathological examinations of vehicle-treated animals with those of animals treated with the test article. The study showed a statistically significant improvement in arthritis scores in animals treated with XTMAB-16 at 20 mg / kg / dose and 40 mg / kg / dose compared to vehicle control-treated animals. This effect was observed at the end of the 29-day treatment period and at the end of the 43-day recovery period (p <.01), as Figure 4 shown. There was no significant difference in arthritis scores between the 20 mg / kg / dose and 40 mg / kg / dose (t-test: p >.05), presumably because these dose levels exceeded the maximum pharmacologic effect dose.

[0491] Example 2: Evaluation of the Minor Pharmacodynamics of XTMAB - 16

[0492] The mechanism of action of XTMAB-16 can involve neutralizing TNFα by binding to soluble or membrane-bound TNFα. Under certain conditions, TNFα binding molecules can mediate complement-dependent cytotoxicity (CDC) and / or antibody-dependent cell-mediated cytotoxicity (ADCC) of cells carrying mTNF. Therefore, immune responses that activate ADCC and CDC can serve as secondary mechanisms of action of XTMAB-16 and may also affect drug safety, and the binding affinities to C1q and the FcγR family, as well as the neonatal Fc receptor (FcRn), may affect the antibody half-life (T 1 / 2 )

[0493] C1q binding

[0494] C1q-ELISA assays were performed using plates coated with C1q and XTMAB-16 binding curves ranging from 4 μg / mL to 2000 μg / mL. However, the C1q binding results using this ELISA method cannot predict the relative activity results of the CDC assay. Since the ELISA for C1q binding was performed at relatively high concentrations, it may be subject to interference. Additional development will be carried out to determine the appropriate conditions and binding curve ranges for the assay.

[0495] Complement-dependent cytotoxicity

[0496] The CDC of XTMAB-16 was assayed using cells expressing mTNFα incubated with human serum as a source of complement factors. The cells were incubated in the presence of human serum (Quidel) and increasing amounts of XTMAB-16 and incubated for 4 hours. The cell density was determined by luminescence after adding the CellTiterGlo reagent. The luminescence signal is proportional to the cell density.

[0497] The CDC activities are presented in Table 10. The XTMAB-16 batches were stable at 2 - 8 °C for 2 months, with CDC activities of 92% to 93%. Similar results were previously achieved, which demonstrated the CDC activities of 4 different XTMAB-16 batches, with the range of CDC activities between 92% and 119% compared to infliximab.

[0498] Table 10: Relative CDC activities of XTMAB-16 DS and DP

[0499] XTMAB - 16 Batches Reference Materials Relative Activity (%) A DP Infliximab 110 A DP XTMAB - 16: B RS 97 ADS XTMAB - 16: B RS 92 A, DS, 1 month, at 2 - 8°C XTMAB - 16: B RS 93 A, DS, 2 months, at 2 - 8°C XTMAB - 16: B RS 92

[0500] Abbreviations: CDC = complement-dependent cytotoxicity; Abbreviation: DP = drug product; DS = drug substance; RS = reference standard.

[0501] Antibody-dependent cell-mediated cytotoxicity

[0502] ADCC assays were performed using isolated peripheral blood mononuclear cells (PBMCs). Target T3-33 cells stably expressing mTNFα (3.75×10 4 cells / well) were incubated with different concentrations of XTMAB-16, and 375,000 signals were proportional to cell death. Compared to infliximab, the ADCC activity of reference standard (RS) lot B was 74% to 79%, and the ADCC activity of lot A was 73% to 113% relative to lot B (Table 11).

[0503] Table 11: Relative ADCC activities of XTMAB-16 DS and DP compared to B

[0504]

[0505]

[0506] Abbreviations: ADCC = antibody-dependent cell-mediated cytotoxicity; DP = drug product; DS = drug substance; RS = reference standard.

[0507] Fc receptor binding

[0508] Receptors that recognize the Fc portion of anti-TNFα antibodies can be expressed on leukocytes and are divided into three groups, which are named: FcγRI, RII, and RIII (also known as CD64 (GenBank: AAI52384.1), CD32 (GenBank: AAI48274.1), and CD16 (NCBI: NP_001373379.1), respectively). FcγRI exhibits high affinity for IgG (10-8 M to 10-9 M) and can bind monomeric IgG. In contrast, FcγRII and FcγRIII can show weaker affinity for monomeric IgG, association constants (K a ≤10-7 M), and can only interact effectively with polymeric immune complexes. FcγRI, RII, and RIII can participate in ADCC.

[0509] Thus, the evaluation of different aspects of XTMAB-16Fc binding via Fc receptors can provide insights into possible conformational changes. Generally, for Fc binding assays, label-free assays using the Octet QK384 system were used to detect the binding kinetics of XTMAB-16, and the resulting signals were converted into affinity parameters by Octet analysis software (Forte'Bio Data Acquisition 9.0 software and Forte'Bio Data Analysis 9.0 software). Two methods were employed for comparative analysis of Fc binding: ELISA for evaluating relative binding and analysis of binding kinetics using the Octet QK384 system and SoftMaxPro software.

[0510] Binding to FcγRI

[0511] The binding kinetics of XTMAB-16 to FcγRI (also known as CD64 (GenBank: AAI52384.1)) were determined using the Octet QK384 system. Briefly, FcγRI was immobilized onto a nickel-coupled biosensor (Ni-NTA). The binding kinetics of XTMAB-16 to FcγRI were analyzed at 7 XTMAB-16 concentrations, and binding was determined by fitting to a 1:1 model. Figure 5A and 5B Sensorgrams are shown in, which show the binding kinetics of XTMAB-16 batches to FcγRI. The binding affinity of XTMAB-16 to FcγRI is presented in Table 12.

[0512] Table 12: Binding affinity of XTMAB-16 to FcγRI

[0513] Batch <![CDATA[K D (nM)]]> XTMAB - 16D DS 0.68 XTMAB - 16B RS 0.63

[0514] Abbreviations: DS = drug substance; K D = affinity constant; RS = reference standard.

[0515]

[0516]

[0517] Abbreviations: DS = drug substance; K D = affinity constant; RS = reference standard.

[0518] The K D of XTMAB-16 batches for FcγRI was in the range of 5 to 9×10-10 M.

[0519] An ELISA was performed to determine the relative binding of XTMAB-16 to the FcγRI receptor using recombinant purified His-tagged FcγRI, which was bound to the ELISA plate via an anti-His tag antibody and incubated with XTMAB-16 or infliximab at 3 ng / mL to 3000 ng / mL. Binding was detected using peroxidase-conjugated F(ab')2 goat anti-human IgG and the HRP chromogenic substrate TMB with a microplate reader set at 450 nm.

[0520] Binding to FcγRIIIa

[0521] ADCC can be initiated through the interaction of the Fc region with FcγRIIIa. FcγRIIIa (CD16) is a 50 kDa to 70 kDa membrane glycoprotein that can be expressed by neutrophils, eosinophils, NK cells, and tissue macrophages. FcγRIII shows weak affinity for monomeric IgG, Ka ≤ 10 7 M -1 , and thus can only interact effectively with polymeric immune complexes.

[0522] To determine the binding kinetics of XTMAB-16 to FcγRIIIa, His-tagged FcγRIIIa was immobilized onto a biosensor that binds to an anti-penta-His (SEQ ID NO:25) antibody. The binding kinetics of XTMAB-16 to FcγRIIIa were analyzed at 7 XTMAB-16 concentrations, and kinetic analysis was performed using the Octet QK384 (Forte'Bio) instrument with a 2:1 heteroligand model fit. The results are shown in Figure 6A and 6B and Table 13.

[0523] Table 13: Binding affinity of XTMAB-16 to FcγRIIIa

[0524] Batch <![CDATA[K D (M)]]> XTMAB - 1DS <![CDATA[6.5×10 -7 <!-- 43 -->]]> XTMAB - 16RS <![CDATA[5.2×10 -7 >

[0525] Abbreviations: DS = drug substance; K D = affinity constant; RS = reference standard

[0526] Steady-state K determined from the binding curves of different XTMAB-16 batches to FcγRIIIa D in the range of 3 to 7 × 10 -7 M.

[0527] An ELISA was performed to evaluate the comparative binding of XTMAB-16 to FcγRIIIa. Relative to infliximab, XTMAB-16 exhibited 79% to 92% binding to FcγRIIIa. This lower binding may be attributed to a lower level of fucosylated glycans in the XTMAB-16 batches compared to infliximab. Fucosylated IgG1 may have a lower affinity for FcγRIIIa, which may be due to steric hindrance caused by fucose.

[0528] Binding to FcγRIIb

[0529] To determine the binding kinetics of XTMAB-16 to FcγRIIb, FcγRIIb was immobilized onto a nickel-coupled biosensor (Ni-NTA), and XTMAB-16 binding was analyzed at 6 concentrations. Kinetic analysis was performed using an Octet QK384 (Forte'bio) instrument with a 1:2 heterologous ligand model fit. The results are summarized in Figure 7A 、 Figure 7B and Table 14.

[0530] Table 14: Binding affinity of XTMAB 16 to FcγRIIb

[0531] Batch <![CDATA[K D (M)]]> XTMAB - 16DS <![CDATA[5.6×10 -7 > XTMAB - 16RS <![CDATA[6.4×10 -7 >

[0532] Abbreviations: DS = drug substance; K D = affinity constant; RS = reference standard.

[0533] An ELISA assay performed to compare XTMAB-16 binding with infliximab binding showed that relative to infliximab, the binding of XTMAB-16 to FcγRIIb was 97% to 114%.

[0534] Binding to FcRn

[0535] The binding of IgG to FcRn can protect the antibody from degradation, thereby targeting the antibody to the endocytic salvage pathway, which returns the antibody to the serum, thereby prolonging the T 1 / 2 of IgG. The binding of IgG and FcRn can be pH-dependent: at pH 6, FcRn binds to IgG with nanomolar affinity, while at pH 7, the binding of IgG to FcRn is undetectable.

[0536] Determine the binding kinetics of XTMAB-16 to FcRn through the Fc domain. Fix His-tagged FcRn to a biosensor coated with anti-penta-His (SEQ ID NO:25) antibody. Analyze the binding kinetics of XTMAB-16 to FcRn at 7 XTMAB-16 concentrations. Use an Octet QK384 (Forte'bio) instrument to fit the kinetic analysis with a 1:1 heterologous ligand model. The results are shown in Figure 8A and Figure 8B and Table 15.

[0537] Table 15: Binding affinity of XTMAB 16 to FcRn

[0538] Batch <![CDATA[K D (nM)]]> XTMAB - 16D DS 4.7 XTMAB - 16D DS, 1 month, at 2 - 8°C 3.4 XTMAB - 16D DS, 2 months, at 2 - 8°C 3.5 to 4.5 XTMAB - 16B RS 3.4 to 4.3

[0539] Abbreviations: DS = drug substance; KD = affinity constant; RS = reference standard.

[0540] To determine the similarity to infliximab, the K D determined for the binding affinity of XTMAB-16 batches to FcRn is close to the preliminary target range (4.5 nM to 7 nM).

[0541] Two methods were used to analyze the FcRn binding of XTMAB-16: ELISA for evaluating relative binding and analysis of binding kinetics using an Octet QK384 (Forte'Bio) instrument.

[0542] The ELISA assay used purified biotinylated FcRn bound to the assay plate through streptavidin. The XTMAB-16 curve was from 4.1 μg / mL to 1100 μg / mL and bound to FcRn at pH 6.0. The level of bound XTMAB-16 was determined by HRP-conjugated anti-F(ab')2 antibody detected with TMB at 450 nm. The relative binding affinity of 2 independent experiments was 95% to 96%.

[0543] Use an Octet QK384 (Forte'Bio) instrument and biotinylated recombinant FcRn to determine the binding kinetic parameters KD, association constant (Kassociation), and dissociation constant (Kdissociation) of XTMAB-16. Fix biotinylated FcRn to a streptavidin-coated biosensor. Conduct the kinetics of the binding of XTMAB-16 to FcRn at pH 6.0.

[0544] It was found that a 2:1 heterologous ligand fitting model was applicable to the result analysis, and the results indicated the existence of 2 separate binding sites with different affinities on the FcRn molecule. At similar KD , K 缔合 and K 解离 In the case of, the binding affinities of XTMAB-16 batches E, G, H to FcRn are similar to the affinity of infliximab to said FcRn. At 56-fold K D1 In the case of, the affinity of XTMAB-16 batch F to FcRn is higher than the affinities of other batches to FcRn.

[0545] Example 3: Evaluation of Safety Pharmacology

[0546] Mice were administered a single dose of placebo or 10 mg / kg, 20 mg / kg or 40 mg / kg XTMAB-16 (intravenously infused at a volume of 10 mL / kg) on day 1, and the mice were evaluated before dosing and at 3 hours and 312 hours (day 14) after dosing. Randomization and experimental groups are outlined in Table 16.

[0547] Table 16: Randomization and groups

[0548]

[0549] Abbreviation: CNS = central nervous system.

[0550] *The last 10 (5M / 5F) randomly assigned animals were selected for CNS evaluation.

[0551] In the toxicity / CNS safety pharmacology study, there were no significant differences in the short functional observation battery test parameters between the placebo control group and the XTMAB-16 treatment group at 10 mg / kg, 20 mg / kg, 40 mg / kg, respectively, and before dosing, at 2 - 4 hours and 312 hours after dosing.

[0552] In the respiratory safety pharmacology study, there were no significant differences in the respiratory rate, tidal volume, minute ventilation, enhanced expiratory pause between the placebo control group and the XTMAB-16 treatment group at 10 mg / kg, 20 mg / kg, 40 mg / kg, respectively, and before dosing, at approximately 3 hours and 312 hours after dosing.

[0553] In summary, after a single dose in the range of 10 mg / kg to 40 mg / kg, there is no adverse effect on the CNS or respiratory system of TgTC mice treated with XTMAB-16.

[0554] The cardiovascular safety of XTMAB-16 has not been evaluated in any non-clinical studies because there is no suitable species (dog, monkey, pig, rabbit, ferret, guinea pig) pharmacologically relevant to XTMAB-16. Instead of these evaluations, cardiovascular safety was carefully evaluated in XTMAB-16-101 and no findings were noted.

[0555] In vitro examination of human tissue cross-reactivity with XTMAB-16 was performed in normal and diseased human tissues. The goal of this study was to determine the potential cross-reactivity of XTMAB-16 (i.e., a chimeric (human / mouse) IgG1 mAb against TNFα) using frozen sections from a complete set consisting of normal human tissues as well as human sarcoidosis tissues (1-3 donors (if available) for each tissue) at two concentrations [5 μg / mL and 20 μg / mL].

[0556] TNFα is a pleiotropic cytokine produced by macrophages, monocytes, neutrophils, T cells, NK cells, and fibroblasts. However, cells of the monocyte lineage such as macrophages, astrocytes, microglia, Langerhans cells, Kupffer cells, and alveolar macrophages can be the main producers of TNFα. Both soluble and transmembrane forms of TNFα have been identified. TNFα can be involved in cell proliferation, differentiation, growth, and immune responses.

[0557] In normal human tissues, XTMAB-16 stained the membranes and / or cytoplasm of resident cells, migratory cells, infiltrating cells, and / or intravascular monocytes in the bladder, cervix, esophagus, large intestine, liver (Kupffer cells), lymph nodes, lung, pancreas, placenta (Hofbauer cells), parathyroid gland, salivary gland, skin, small intestine, spleen, stomach, thyroid gland, thymus, tonsil, and uterus as expected. Monocyte staining was weak to strong, or weak to moderate and frequent, occasional to frequent, occasional, rare to occasional, or rare at higher concentrations, and equivalent, decreased, or no staining at lower concentrations.

[0558] In tissues obtained from subjects, monocytes in these tissues were selected as secondary controls because TNFα expression was reported in monocytes in human tonsils, small intestine with Crohn's disease, lungs with sarcoidosis, and lymph nodes with sarcoidosis. The purpose of the secondary control tissues was to show XTMAB-16 specific binding in the presence of relevant cell types. XTMAB-16 stained the membranes and cytoplasm of rare to occasional monocytes in human tonsils weakly to strongly at higher staining concentrations and weakly to strongly and rarely at lower concentrations.

[0559] XTMAB-16 also weakly to strongly stains the membranes and cytoplasm of rare to occasional monocytes in the small intestine of individuals with Crohn's disease, the lungs of individuals with sarcoidosis, and the lymph nodes of individuals with sarcoidosis at higher staining concentrations, and stains moderately to rarely at lower concentrations. No staining was observed in the control tissues on the assay control slides.

[0560] The following human tissues were evaluated in this cross-reactivity study:

[0561] 1. Positive control:

[0562] Cryosections of T3-33 (expressing TNFα) cells

[0563] 2. Negative control:

[0564] Cryosections of Sp2 / 0-Ag14 cells

[0565] 3. Assay control:

[0566] Cryosections of human tonsils (monocytes; 2 donors)

[0567] Cryosections of small intestine from individuals with Crohn's disease (monocytes; 1 donor)

[0568] Cryosections of lungs from individuals with sarcoidosis (monocytes; 1 donor)

[0569] Cryosections of lymph nodes from individuals with sarcoidosis (monocytes; 1 donor)

[0570] The normal human tissue types tested are shown in Table 17.

[0571] Table 17: Normal human tissue types tested (3 donors)

[0572] Adrenal Gland Kidney (glomerulus, tubules) Skin Bladder (urine) Liver Spinal Cord Blood Cells a Lung Spleen Blood Vessels (endothelium) b Lymph Nodes Striated Muscle (skeletal) Bone Marrow Ovary Testis Brain - Cerebellum Pancreas Thymus Brain – Cerebral Cortex Parathyroid Gland Thyroid Gland Breast (mammary gland) Peripheral Nerve Tonsil Eye Pituitary Gland Ureter Fallopian Tube / Oviduct Placenta Uterus - Cervix Gastrointestinal (GI) Tract c Prostate Uterus - Endometrium Heart Salivary Gland

[0573] a Evaluated by peripheral blood smear.

[0574] b Evaluated by all tissues present.

[0575] c Including esophagus, large intestine / colon, small intestine, and stomach (including underlying smooth muscle).

[0576] This immunohistochemical evaluation provides support related to the mechanism of XTMAB-16 as a TNFα blocker. As previously noted, TNFα is a pleiotropic cytokine produced by macrophages, monocytes, neutrophils, T cells, NK cells, and fibroblasts. Alveolar macrophage-derived TNFα is involved in the induction and maintenance of granulomas in sarcoidosis, and high levels of TNFα (and high levels of TNFα released from alveolar macrophages) appear to be associated with disease progression. Similar inflammatory mechanisms may underlie sarcoidosis involvement in other organs such as the skin.

[0577] In this study, cross-reactivity of human tissues with XTMAB-16 was expected for both normal and diseased samples. Thus, these immunohistochemical results support the use of XTMAB-16 in subjects with sarcoidosis and are not expected to cause toxicity in tissues not affected by sarcoidosis.

[0578] Example 4: Evaluation of the Pharmacokinetics, Absorption, Distribution, Metabolism, and Excretion of XTMAB - 16

[0579] Absorption and Pharmacokinetics

[0580] In a 3-week non-GLP study conducted in ICR mice, the tmax of a single intravenous administration of XTMAB-16 at 60 mg / kg was 5 minutes. XTMAB-16 administered intravenously weekly showed a dose-dependent relationship, where, respectively, for doses of 30 mg / kg, 40 mg / kg, 60 mg / kg, and 100 mg / kg, the mean plasma concentration at 15 minutes after the third weekly administration was determined to be 512.88 μg / mL, 889.42 μg / mL, 1598.13 μg / mL, and 1976.38 μg / mL.

[0581] According to a 6-week GLP toxicology and TK study conducted in TgTC mice, Cmax and AUC0-∞ increased in a dose-dependent manner after a single dose. After a single intravenous administration, respectively, for doses of 10 mg / kg, 20 mg / kg, and 40 mg / kg, Cmax was 174.8 μg / mL, 354.2 μg / mL, and 791.0 μg / mL. Respectively, for the 10 mg / kg, 20 mg / kg, and 40 mg / kg groups, the AUC0-∞ after a single-dose administration showed linear values of 31755.5 μg / mL*h, 74776.5 μg / mL*h, and 144168.4 μg / mL*h. For all doses, tmax was 0.08 h, and respectively, for the 10 mg / kg and 40 mg / kg doses, the t1 / 2 of XTMAB-16 was in the range between 226.9 h (9.45 days) and 327.2 h (13.63 days).

[0582] From a 13-week non-clinical GLP toxicology study (Study 14042-20005) in TgTC mice, repeated TK parameters were characterized after 6 of 7 bi-weekly intravenous injections of XTMAB-16 at 10 mg / kg, 20 mg / kg, and 40 mg / kg. After 6 administrations of the 40 mg / kg intravenous injection, Cmax was 1104.9 μg / mL, AUC0-∞ was 624865.9 μg*h / mL, and AUC0-τ was 124878.7 μg*h / mL. In the dose range of 10 mg / kg to 40 mg / kg of XTMAB-16, systemic exposure increased in a dose-proportional manner with respect to Cmax and AUC0-∞. In TgTC mice, after 6 repeated bi-weekly intravenous injections of 10 mg / kg, 20 mg / kg, and 40 mg / kg XTMAB-16, all serum samples were negative for ADA up to Day 83.

[0583] Single-dose pharmacokinetics

[0584] In Study 14042-20003, in ICR mice, initial single-dose PK was examined at the 50 mg / kg XTMAB-16 intravenous dosing level. On Day 1, a total of 4 animals (2 animals / sex) were injected with 50 mg / kg XTMAB-16, and plasma concentrations were evaluated at 2 minutes, 5 minutes, 10 minutes, and 15 minutes post-dose. Mean plasma concentrations are summarized in Table 18. Based on the time points evaluated in this study, tmax was determined to be 5 minutes post-dose.

[0585] Table 18: Mean XTMAB-16 Plasma Concentrations after 50 mg / kg Intravenous Dosing in ICR Mice

[0586] Time After Administration 2 minutes 5 minutes 10 minutes 15 minutes Average Plasma Concentration 1057.92 μg / mL 1077.23 μg / mL 723.23 μg / mL 822.86 μg / mL

[0587] Abbreviations: ICR = Institute of Cancer Research; IV = intravenous.

[0588] Examine single-dose PK in TgTC mice. Blood samples were collected before dosing and at 0.083 h, 8 h, 24 h, 48 h, 96 h, 168 h, 336 h, 672 h, 840 h, and 1008 h after dosing on Day 1, and the blood samples were processed into serum. After single intravenous administration of 10 mg / kg, 20 mg / kg, or 40 mg / kg XTMAB-16, serum concentrations were determined, and the major PK parameters are summarized in Table 19. Data for both genders were combined (2 animals / gender / time point, except for the baseline, which was 1 animal / gender / time point). In the dose range of 10 mg / kg to 40 mg / kg, a single dose of XTMAB-16 caused a dose-proportional increase in Cmax, AUC0-τ, and AUC0-∞. At 0.08 h, across doses, Tmax was consistent, and across doses of 0.3 mL / h / kg, CL was consistent.

[0589] Table 19: Single-dose XTMAB-16 Pharmacokinetic Parameters in TgTC Mice

[0590]

[0591] Abbreviations: AUC = area under the curve; CL = clearance; LLOQ = lower limit of quantification; MRT = mean residence time; TgTC = hTNF knock-in transgenic; t1 / 2 = apparent terminal half-life; tmax = time to reach the maximum observed concentration; Vz = volume of distribution at the terminal phase.

[0592] LLOQ: 31.3 ng / mL

[0593] The XTMAB-16 serum concentration vs. time profile is shown in Figure 9 below.

[0594] At doses of XTMAB-16 of 10 mg / kg, 20 mg / kg, and 40 mg / kg, systemic exposure increased proportionally in terms of mean C 最大值 and AUC, as shown in Figure 10 and Figure 11 below.

[0595] Repeated-dose Pharmacokinetics

[0596] In a 3-week exploratory toxicity study in ICR mice, the repeated-dose PK of animals administered 60 mg / kg XTMAB-16 by intraperitoneal (IP) injection and 30 mg / kg, 40 mg / kg, 60 mg / kg, and 100 mg / kg XTMAB-16 by intravenous injection was determined. For animals administered by IP injection, 4 animals (2 males / 2 females) received a dose of 60 mg / kg on Day 1, Day 8, and Day 15, and samples for PK were collected on Day 15 before dosing and at 1 hour, 2 hours, and 3 hours post-dose (N = 1 animal / time point). Plasma concentrations of XTMAB-16 are provided in Table 20. Based on the available time points, the tmax for IP injection in this study was determined to be ≥3 hours.

[0597] Table 20: Plasma Concentrations of XTMAB-16 on Day 15 after Administration of 60 mg / kg on Day 1, Day 8, and Day 15 (N = 1 animal / time point) in ICR Mice

[0598] Time Before Administration 1 hour After Administration 2 hours After Administration 3 hours After Administration Average Plasma Concentration 332.00 μg / mL 696.12 μg / mL 587.44 μg / mL 952.45 μg / mL

[0599] For animals administered by intravenous injection, 4 animals (2 / sex / dose group) in each dose group received intravenous injections of 30 mg / kg, 40 mg / kg, 60 mg / kg, and 100 mg / kg XTMAB-16 on Day 1, Day 8, and Day 15, and samples for PK were collected on Day 15 before dosing and 15 minutes post-dose. Plasma concentrations at 15 minutes post-dose on Day 15 (n = 1 - 2 mice / time point) showed a dose-related increase in concentration, with an average of 512.88 μg / mL at the 30 mg / kg / dose, 889.42 μg / mL at the 40 mg / kg / dose, 1598.13 μg / mL at the 60 mg / kg / dose, and 1976.38 μg / mL at the 100 mg / kg / dose. Pre-dose (trough) plasma concentrations on Day 15 (n = 1 mouse / time point) also showed a dose-related increase, being 322.02 μg / mL at the 30 mg / kg / dose, 353.50 μg / mL at the 40 mg / kg / dose, 479.36 μg / mL at the 60 mg / kg / dose, and 581.84 μg / mL at the 100 mg / kg / dose. The pre-dose data indicate that the analyte had not been completely eliminated from the systemic circulation within the 1-week period between the second and third doses. The dose-response data are summarized in Figure 12 in.

[0600] Pharmacokinetic (PK) samples were collected from 7 animals administered intravenously that died on Day 8 and 1 male (plasma concentration of 276.15 μg / mL) that died on Day 1 and was replaced at 60 mg / kg. At doses of 40 mg / kg / dose to 60 mg / kg / dose, the Day 8 concentrations of XTMAB-16 in these mice were generally comparable (at 40 mg / kg / dose, 624.49 μg / mL; at 50 mg / kg / dose, 359.9 μg / mL to 614.40 μg / mL; and at 60 mg / kg / dose, 430.99 μg / mL to 455.01 μg / mL), and increased at 100 mg / kg / dose (805.53 μg / mL).

[0601] Repeated-dose TK was examined in a completed 13-week TgTC mouse toxicology study. A total of 40 animals per group (20 animals / sex / dose group) received doses of 10 mg / kg, 20 mg / kg, or 40 mg / kg of XTMAB-16 every two weeks on Days 1, 15, 29, 43, 57, and 71. Repeated TK parameters were characterized after every two-week intravenous injection for 6 of the 7 every-two-week intravenous injections of XTMAB-16 at 10 mg / kg, 20 mg / kg, and 40 mg / kg. After 6 administrations of the 40 mg / kg intravenous injection, C 最大值 was 1104.9 μg / mL, AUC0-∞ was 624865.9 μg*h / mL, and AUC0-τ was 124878.7 μg*h / mL. Systemic exposure increased in a dose-proportional manner with respect to Cmax and AUC0-∞ in the dose range of 10 mg / kg to 40 mg / kg of XTMAB-16.

[0602] PK-related safety margin

[0603] In the 13-week toxicology study, mg / kg scaling using the 40 mg / kg TgTC mouse NOAEL provided safety margins of 5-fold and 10-fold for the 2 mg / kg and 4 mg / kg human dose groups, respectively. Projected human safety margins were also calculated by comparing the NOAEL exposure in the toxicology species (TgTC mice dosed at 40 mg / kg) with projected clinical exposure. Using TK data (Table 21) generated from a 6-week GLP toxicology study in TgTC mice, a 2-compartment model with first-order absorption and elimination rates using a hybrid ratio error model was developed for XTMAB-16. The model was scaled allometrically relative to humans using published allometric exponents. The allometrically scaled human model was then used to simulate the planned FIH dose of XTMAB-16 for a 70 kg human.

[0604] The predicted human dose is then compared with C 最大值 and AUC 0-∞ calculated from the 13-week NOAEL to generate an exposure-based safety margin. Using a model established from a 6-week toxicology study comparing the AUC 0-∞ (624865.9 hour·mg / mL) of the 40 mg / kg group of XT-MAB16 administered in the TgTC model toxicology study with the estimated AUC of a 1 mg / kg dose (1400 hour·mg / mL) in humans 0-∞ the safety margin for the starting dose is approximately 45-fold. Comparing the Cmax (1104.9 mg / mL) of XTMAB-16 at the 40 mg / kg dose in TgTC with the estimated Cmax (47.8 mg / mL) of a 2 mg / kg dose in humans, the safety margin for the starting dose is approximately 23-fold.

[0605] Table 21: Human safety margins compared with Cmax and AUC calculated from TgTC mice

[0606]

[0607] Abbreviations: AUC = area under the curve; Cmax = maximum observed concentration; TgTC = hTNF knock-in transgenic.

[0608] Excretion

[0609] The clearance (CL) of XTMAB-16 after a single intravenous injection of 10 mg / kg, 20 mg / kg, or 40 mg / kg in TgTC mice was calculated to be 0.3 mL / hour / kg. At steady state, the CL after 6 administrations in the 13-week toxicology study was calculated to be 0.12.

[0610] Example 5: Evaluation of the Toxicology of XTMAB - 16

[0611] In a 3-week non-GLP, exploratory toxicology study in ICR mice, 25 animals were assigned to receive weekly intravenous administrations (days 1, 8, and 15) of XTMAB-16 at 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, or 100 mg / kg. Eight of the 21 animals died within 1 hour after dosing on day 8 as follows: at 30 mg / kg, 0 / 4; at 40 mg / kg, 1 / 4; at 50 mg / kg, 3 / 4; at 60 mg / kg, 2 / 4; and at 100 mg / kg, 1 / 4. After weekly intravenous administrations, 40 mg / kg was determined to be the highest dose to be used in subsequent non-clinical studies.

[0612] In a 6-week GLP, TgTC mouse toxicology study (4-week treatment period, 2-week recovery period), 60 animals (10 animals / sex / dose level) received repeated intravenous administration of XTMAB-16 at 0 mg / kg, 20 mg / kg, or 40 mg / kg on Day 1, Day 15, and Day 29. Another 132 animals received a single intravenous administration of XTMAB-16 at a dose of 10 mg / kg, 20 mg / kg, or 40 mg / kg on Day 1 and were assigned to the TK satellite group. No animals died in the toxicology group. In the TK satellite group, 3 animals died: 1 male died 7 days after dosing (10 mg / kg group), and 2 females died 15 days after dosing (20 mg / kg group). These deaths were considered unrelated to XTMAB-16 because they occurred 1 to 2 weeks after dosing and there was no dose-response relationship. After 3 bi-weekly intravenous injections in TgTC mice, the NOAEL of XTMAB-16 was considered to be 40 mg / kg / dose.

[0613] In a 13-week non-clinical GLP toxicology study conducted in TgTC mice, after 7 bi-weekly intravenous injections in TgTC animals in the toxicity study, XTMAB-16 was well tolerated up to 40 mg / kg, and no significant test article-related clinical and pathological abnormalities were observed. There were disease model-related abnormalities. Animal deaths were observed at all doses up to and including 40 mg / kg; however, these deaths were not dose- or time-dependent, and there were no pathological changes that would allow determination of the cause of death. Overall, animals dosed after the first dose did not die, and all animals dosed with XTMAB-16 survived for at least 4 weeks after the first dose, with most surviving 9 weeks or longer. Given the death of 4 untreated spare mice, the deaths were not considered to be test article-related. The no-observed-adverse-effect level (NOAEL) of XTMAB-16 was 40 mg / kg.

[0614] Example 6: Additional Pharmacokinetic Studies of XTMAB - 16

[0615] For both the 2 mg / kg group and the 4 mg / kg group, the overall mean XTMAB-16 concentrations were presented against nominal time on linear and semi-logarithmic profiles and were evaluated in non-sarcoidosis subjects ( Figure 13A and 13B ). The PK profiles of the 2 mg / kg group and the 4 mg / kg group showed a rapid increase in plasma concentration during infusion, with t 最大值 between 2 and 3 hours, and a slow elimination in a two-phase disposition manner, with overall geometric mean half-lives of 7.8 hours and 9.6 hours in the 2 mg / kg group and the 4 mg / kg group, respectively.

[0616] For serum concentration-time profiles aggregated by treatment and ADA incidence, in the two XTMAB-16 groups, the mean profiles of subjects showing a positive ADA / Nab response exhibited a more rapid elimination from the systemic circulation compared to the profiles of subjects showing a negative ADA / Nab response ( Figure 14A and 14B ).

[0617] Separately, for XTMAB-16 in all subjects receiving a single dose of 2 mg / kg and 4 mg / kg, the geometric mean Cmax (geometric CV%) of XTMAB-16 was 63.52 μg / mL (15%) and 129.1 μg / mL (18.8%), respectively. Separately, for 2 mg / kg and 4 mg / kg, the corresponding geometric means (geometric CV%) of systemic XTMAB-16 exposure (AUC0-last) were 483.6 day*μg / mL (44.3%) and 1203 day*μg / mL (20.3%), respectively. In addition, for both groups, AUC0-∞ was comparable to AUC0-last Table 22.

[0618] Table 22: Summary of XTMAB-16 serum pharmacokinetic parameters by treatment in ADA / nAb positive and overall pooled subjects in Study XTMAB-16-101

[0619]

[0620]

[0621]

[0622]

[0623] Abbreviations: AUC0-last = AUC from time 0 to the time of the last quantifiable concentration; AUC0-∞ = AUC extrapolated from time 0 to infinite time; CL = clearance; Cmax = maximum concentration; CT = concentration at the end of the infusion time (i.e., 2 hours); CV = coefficient of variation; DNAUC0-last = dose-normalized AUC0-last; DNAUC0-∞ = dose-normalized AUC0-∞; DNCmax = dose-normalized Cmax; Geo = geometric; Max = maximum; Min = minimum;

[0624] N = The number of subjects in the pharmacokinetic population undergoing the corresponding treatment; n = The number of observations; SD = Standard deviation; tmax = The time of Cmax; t1 / 2 = Terminal half-life; Vz = Volume of distribution at the terminal stage; Vss = Volume of distribution at steady state; MRT = Mean residence time; λz = Terminal elimination rate constant

[0625] The statistical analysis of the XTMAB-16 dose proportionality using the power model is presented in Table 28. In this analysis, the slope of Cmax (1.02) is close to 1.00, and the 95% CI [0.79, 1.26] almost falls within the dose proportionality interval [0.8, 1.25]. However, the slope values of the PK parameters AUC0-last (1.31) and AUC0-∞ (1.10) are greater than 1.00, and the upper limits of the corresponding 95% CI from 2 mg / kg to 4 mg / kg show an interval greater than the usual dose proportionality interval [0.8, 1.25], thus showing a greater-than-dose-proportional increase in AUC.

[0626] Based on the dose-normalized PK parameters, the dose-normalized Cmax (DNCmax) and dose-normalized AUC0-∞ (DNAUC0-∞) between the two groups are almost exactly the same. However, there is some difference in DNAUC0-last in the 2 mg / kg group compared to the 4 mg / kg group.

[0627] Table 23: Study XTMAB-16-101: Statistical analysis of dose proportionality for evaluating the serum pharmacokinetic parameters of XTMAB-16

[0628] Parameter (unit) n μ β (SE) 95% CI of β Cmax (μg / mL) 19 3.442 1.023(0.111) 0.79；1.26 AUC0-last (day * μg / mL) 18 5.270 1.315(0.225) 0.84；1.79 AUC0-∞ (day * μg / mL) 17 5.581 1.104(0.189) 0.70；1.51

[0629] Abbreviations: N = The number of observations; μ = Intercept; β = Slope; SE = Standard error of the slope; CI = Confidence interval.

[0630] The power model is used to evaluate the dose proportionality.

[0631] Model: The fitted linear model is log(Y) = μ + β * log(dose), where Y is the PK parameter.

[0632] Population pharmacokinetic analysis

[0633] Based on the data collected in Study XTMAB-16-101, population PK analysis and simulation of XTMAB-16 in non-sarcoidosis adult subjects were performed.

[0634] The PK analysis dataset included 277 PK observations from 19 subjects. The number of subjects, measurable observations, and observations below the limit of quantification (BLQ) across studies were summarized by study in Table 24. Concentration-time profiles with intensive sampling were available for all 19 subjects at 2 mg / kg and 4 mg / kg.

[0635] Serum XTMAB-16 samples with quantifiable / reported concentration values were not excluded from the analysis. Individual subjects were not excluded from the analysis.

[0636] Table 24: Plasma Concentrations in the PK Analysis Dataset

[0637]

[0638] Abbreviations: BLQ = below the limit of quantification; PK = pharmacokinetics. Source: Modeling and Simulation Report

[0639] NCA was performed to provide initial estimates for the population PK analysis of XMTAB-16. Actual time was used in the analysis. BLQ values were set to 0 before Tmax and missing after Tmax. PK parameters were summarized by dose level and ADA status in Table 25. Exposure was reduced in ADA-positive subjects compared to ADA-negative individuals in terms of AUC.

[0640] Table 25: Summary Statistics of Serum Pharmacokinetic Parameters of XTMAB-16 after Single Intravenous Administration

[0641]

[0642]

[0643] Abbreviations: AUC∞ = area under the serum concentration-time curve; AUC last = area under the serum concentration-time curve from time 0 to the last measurable concentration; Ceoi = concentration at the end of infusion; Cmax = maximum serum concentration; CL = clearance; D = dose; MRT∞ = mean residence time to infinity; NC = not calculated; T1 / 2 = half-life; Tmax = time of the observed maximum concentration; T last = time of the last quantifiable plasma concentration; Vss = volume of distribution at steady state; Vz = volume of distribution at the terminal phase, parameters are presented as mean (CV%); n for all except Tmax and T last presents median (max - min); n Ceoi was set to the measured serum XTMAB-16 concentration at 2-hour nominal time.

[0644] Source: Modeling and Simulation Report

[0645] Using the final base population PK model, selected covariates were evaluated in a univariate manner, and the covariates included:

[0646] 1. In terms of CL, body weight, sample-level ADA, subject-level ADA, and subject-level nAb

[0647] 2. In terms of the central volume of distribution (V), body weight and subject-level ADA

[0648] 3. In terms of the peripheral volume of distribution (V2), body weight

[0649] 4. It was found that weight and subject-level ADA had a significant impact on the PK of XTMAB-16.

[0650] The final population PK model of XTMAB-16 is a two-compartment model, which includes linear elimination and inter-individual variability in terms of CL, central volume of distribution, and peripheral volume of distribution. Body weight was included as a covariate in the volume parameters, and ADA status was included in terms of CL. Based on the final model, CL was 0.009 L / h, the volume of distribution at steady state (Vss) was 3.02 L, and the estimated t1 / 2 for a typical individual (where the weight was 72.4 kg) was 243.3 hours (194.0 hours in the case of ADA-positive subjects). The parameters of the final XTMAB-16 population PK model were estimated with good precision; the percentage of relative standard error of PK parameters and random effects was <23%. The goodness-of-fit curve plot indicated reasonable fitting of the data. The predicted concentrations for individuals and the population showed good correlation with the observed data. Regarding the PK of XTMAB-16, no trend with respect to age was detected, and the residual trend with respect to gender was small. Due to the small sample size, the effects of race and ethnicity could not be strongly estimated; however, the box plot of ethnicity indicated an overlap between Hispanic / Latino subjects and non-Hispanic / non-Latino subjects.

[0651] Example 7: Evaluation of XTMAB-16 in in vitro models of sarcoidosis and granulomatous diseases

[0652] This example evaluated the activity of XTMAB-16 in an in vitro model of granuloma formation established using peripheral blood mononuclear cells from subjects with active sarcoidosis to determine a potentially effective dose range. Data obtained from the first-in-human study of XTMAB-16 (NCT04971395) were used to develop a population pharmacokinetic (PPK) model for characterizing the pharmacokinetics (PK) of XTMAB-16. Model simulations were performed to evaluate the sources of PK variability and to predict lung interstitial exposure based on the concentrations in the in vitro granuloma model.

[0653] Dose levels of XTMAB-16 of 2 mg / kg and 4 mg / kg, once every 2 weeks (Q2W) or once every 4 weeks (Q4W) for up to 12 weeks were supported by the following: data from non-clinical in vitro secondary pharmacology; a Phase 1 clinical study; and a PPK model used to guide dose level and frequency assumptions. In an in vitro granuloma model, respectively, at half-maximal inhibitory concentrations (IC50) of 5.2 μg / mL and 3.5 μg / mL, XTMAB-16 inhibited granuloma formation and blocked interleukin-1 (IL-1) secretion. The mean interstitial lung concentration following administration of 2 mg / kg or 4 mg / kg Q2W or Q4W was expected to exceed the in vitro IC50 concentration.

[0654] The data presented herein provide the rationale for the dose selection of XTMAB-16 in subjects with sarcoidosis in Example 8.

[0655] Materials and Methods

[0656] An in vitro sarcoidosis model that generates granuloma formation was used to closely replicate the molecular characteristics of granulomas in diseased sarcoid tissue (8-13). Peripheral blood mononuclear cells (PBMC) were isolated from subjects with active pulmonary sarcoidosis with lung and / or lymph node involvement who were untreated for tuberculosis (TB) (negative purified protein derivative [PPD] skin test and / or Quantiferon Gold test; N = 10), non-smokers, and not treated with potent immunosuppressants (e.g., methotrexate, azathioprine, prednisone, anti-TNF antibody) within 3 months, and cultured in standard 24-well plates. The PMBC were exposed to uncoated beads (UNC) or beads coated with the Mycobacterium tuberculosis (M. tb) antigen PPD for 7 days. Sarcoid-like PBMC were cultured for 30 minutes in the presence or absence of inhibitor pretreatment (XTMAB-16 or prednisone) (within the dose range that has been shown to inhibit the inflammatory response of cultured human PBMC in previous studies), followed by exposure to PPD-coated beads ( Figure 15 ). Cells were imaged 7 days after treatment. Quantitative analysis of granuloma size / frequency was performed (using MIPAR image processing software). The area fraction for each treatment group was calculated based on at least 4 independent images, and the area fraction was defined as the percentage of the cumulative granuloma area to the total area of the image. After imaging, supernatant samples were collected and extracellular cytokine release was quantified using an enzyme-linked immunosorbent assay.

[0657] Granuloma formation following pretreatment with XTMAB-16 (1 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, and 40 μg / mL) or prednisone (1 μM and 10 μM; 0.358 μg / mL and 3.58 μg / mL, respectively; 30 minutes prior to PPD-coated bead exposure) was compared to granuloma formation following treatment with PPD-coated beads alone. UNC served as a comparable negative control.

[0658] The model study was conducted under ClinicalTrials.gov identifier NCT01857401, which is hereby incorporated by reference.

[0659] Phase 1 XTMAB-16 Study

[0660] The Phase 1 study of XTMAB-16, XTMAB-16-101 (NCT04971395), was a randomized, double-blind, placebo-controlled first-in-human study to evaluate the safety, tolerability, PK, and immunogenicity profile of a single intravenous infusion of XTMAB-16 in non-sarcoidosis, non-smoking adults. XTMAB-16 was administered as an intravenous infusion over a 2-hour period at a dose of 2 mg / kg or 4 mg / kg. Based on the established no-observed-adverse-effect level (NOAEL) (40 mg / kg) of XTMAB-16 in the non-clinical program, the maximum clinical starting dose was 4 mg / kg.

[0661] Inclusion criteria included adult men or women without sarcoidosis; age 18 to 45 years (inclusive); weight between 45 kg and 100 kg and body mass index (BMI) between 18.0 kg / m 2 and 30.0 kg / m 2 (inclusive); and having clinically acceptable clinical laboratory values and electrocardiogram results. Exclusion criteria included receipt of any investigational compound within 90 days prior to dosing and any major disease. A total of 25 non-sarcoidosis adult subjects were enrolled and assigned to two treatment groups (2 mg / kg or 4 mg / kg) or placebo. The mean age (minimum, maximum) of the subjects was 32 years (19 years, 45 years); most were female (N = 16, 64%), black or African American (N = 20, 80%), and not Hispanic or Latino (N = 19, 76%); and the mean BMI (minimum, maximum) was 25.84 (20.9, 29.8) (Table 26).

[0662] Table 26: Demographic Characteristics: Safety Set

[0663]

[0664]

[0665] Pharmacokinetic analysis

[0666] PK analysis was performed on the data collected in XTMAB-16-101. For population PK (PPK) analysis, the dataset included information on subject identification, the date and time of XTMAB-16 administration, the calibration date and time of PK sample collection and the actual date and time, the concentration of XTMAB-16 in serum, and data on relevant covariates, including weight, age, height, BMI, dose, subject-level anti-drug antibody (ADA), sample-level ADA, gender, race, and ethnicity. For the purposes of this analysis, samples below the limit of quantification (BLQ) were set to missing.

[0667] Using NLME (v 8.4, Certara) was used for PPK model development. Based on a visual inspection of the concentration-time profiles, a two-compartment model was used as the starting point for this modeling exercise.

[0668] Covariate analysis: After selecting the base PPK model, the relationship between the covariates of XTMAB-16 and the PK parameters was first explored graphically to obtain information on covariates that might affect the parameters of interest and to guide covariate analysis. The correlation between covariates was considered during the covariate selection process (i.e., in cases where two covariates were assumed or graphically shown to be correlated, one of the two covariates would be selected in the formal covariate model testing strategy).

[0669] Model evaluation: Model evaluation was based on standard model diagnostics and goodness-of-fit criteria (e.g., accuracy of parameter estimation [i.e., 95% confidence interval excluding 0], successful model convergence) and was performed by looking at relevant graphical representations of goodness-of-fit (e.g., fitted and observed concentrations vs. time, conditional weighted residuals vs. time).

[0670] Internal quantification of the final model was performed by predictive-corrected visual predictive check (pcVPC). The pcVPC was constructed using the final model estimates, and the pcVPC involved 1000 replicates of simulated concentration-time profiles, followed by normalization based on typical population predictions to help account for differences in study design.

[0671] After successful evaluation of the final PPK model, individual estimates of PK and exposure parameters were derived by posterior Bayesian estimation, and the individual estimates were compared to the PK parameters previously determined by non-compartmental analysis.

[0672] Simulation: At 2 mg / kg and 4 mg / kg, a once-every-two-weeks (Q2W) and a once-every-four-weeks (Q4W) regimen were simulated over a 16-week period. In the first 12 weeks, a total of 6 doses were administered in the Q2W regimen and 3 doses were administered in the Q4W regimen. Monte-Carlo simulation was performed, whereby the PPK model parameters for each virtual subject (N = 500) were obtained from the sampling through the final PPK variance-covariance matrix. To account for body weight, body weight was randomly assigned from a uniform distribution of 50 - 100 kg. It was assumed that the infusion time of XTMAB-16 for each virtual subject was 2 hours. The ADA status was included as a predictor in the simulation.

[0673] To account for the concentration in lung tissue, the antibody biodistribution coefficient of lung tissue (14.9%) was used to estimate the interstitial lung concentration of XTMAB-16. Then, the exposure parameters in the lung and serum after the first and last doses were calculated for a typical individual with a body weight of 70 kg and in an ADA-negative state, and for illustrative purposes, the exposure parameters were presented in tabular form.

[0674] For all groups, 2 subjects (1 assigned to XTMAB-16 and 1 assigned to placebo; sentinel group) were dosed at least 48 hours before the other subjects in the same group. After a blinded review of the available safety and laboratory data for the sentinel group, and in the absence of an identified significant safety signal, the remaining subjects in the same group were dosed. Subjects within the group were dosed at least 1 hour apart.

[0675] Subjects in the 4 mg / kg group received XTMAB-16 after the 2 mg / kg group was completed and at least 14 to 21 days after the clinical evaluation of the subjects in the 2 mg / kg dose group.

[0676] After a screening period of up to 28 days, subjects were allowed to enter the Clinical Research Unit (CRU) on Day - 2 for COVID-19 testing and baseline procedures. After eligibility was confirmed, subjects were randomly assigned to treatment (1:1 XTMAB-16 to placebo for the sentinel group and 3:1 for the remaining subjects), and the study drug was administered on Day 1. Subjects remained in the CRU until Day 8 for PK, biomarker, anti-drug antibody (ADA), and safety assessments. After all procedures were completed on Day 8, subjects were discharged on the same day. Subjects returned to the CRU on Days 15, 29, 43, 57, and 71.

[0677] Safety assessments included adverse events, safety laboratory assessments, vital signs, electrocardiograms, and physical examinations. During the course of the study, PK collection and analysis were also completed in a similar manner.

[0678] Results

[0679] Study of the activity of XTMAB-16 in an in vitro granuloma model of sarcoidosis: Demographic characteristics of the subjects included in the in vitro study are shown in Table 27. The designation of high responders or low responders indicates the response to PPD-coated beads in terms of in vitro granuloma formation.

[0680] Table 27: Demographics of the subjects included in the in vitro study

[0681]

[0682] Representative micrographs of granuloma-like cell aggregates formed 7 days after pretreatment with XTMAB-16 or prednisone are shown in Figure 16 after incubation of high responder sarcoidosis PBMC with UNC or PPD-coated beads. Pretreatment with XTMAB-16 significantly attenuated granuloma formation in the in vitro model in a dose-dependent manner in the range of 1 μg / mL to 40 μg / mL. The effect of higher doses of XTMAB-16 in the in vitro granuloma model was comparable to the effect of pretreatment with prednisone (1 μM to 10 μM; 0.358 μg / mL to 3.58 μg / mL).

[0683] Pretreatment with XTMAB-16 for 30 minutes had a significant dose-response effect on inhibiting granuloma formation 7 days after treatment with PPD in the in vitro granuloma model, as calculated by area fraction ( Figures 17A - 17C ). For comparison, a similar response to prednisone pretreatment is shown.

[0684] Pretreatment with XTMAB-16 for 30 minutes had a significant dose-response effect on inhibiting granuloma cytokine release 7 days after treatment with PPD in the in vitro granuloma model ( Figures 18A - 18C ). For comparison, a similar response to prednisone pretreatment is shown.

[0685] Separate statistical analyses were performed to evaluate the half-maximal inhibitory concentration (IC50) of XTMAB-16 for MIPAR area fraction and the interleukin (IL)-1 concentration in the supernatant of the in vitro granuloma assay for all subject samples ( Figure 19A and 19B)。The 95% CI of the IC50 was not estimated because the software was unable to calculate the complete confidence interval. The IC50 for the reduction in area fraction was 5.197 μg / mL, and the IC50 for the reduction in IL-16β was 3.465 μg / mL.

[0686] Among subjects, the in vitro granuloma response was variable. Despite this variability, the dose response to XTMAB-16 in vitro was consistent, as reflected by the objective imaging to evaluate the granuloma area. The results were comparable to those of prednisone. In this in vitro model, IL-1b (rather than TNFα or IFNy) best reflected the granuloma response and could therefore be a biomarker for future clinical studies of sarcoidosis. Based on the in vitro dose response to XTMB-16, first-in-human study data were used for PPK modeling and simulation to inform the ongoing clinical development of XTMAB-16 for the treatment of sarcoidosis.

[0687] PPK Modeling

[0688] PK measurement results: The PK analysis dataset included 277 PK observations from 19 subjects. The measurable PK observations were 226 (81.6%); 19 (6.9%) were pre-dose BLQ samples, and 32 (12.4%) were post-dose BLQ samples. The BLQ observations after the first dose (“post-treatment BLQ”) accounted for 12.4% of all post-dose observations. Serum XTMAB-16 samples with quantifiable / reported concentration values were not excluded from the analysis. Individual subjects were not excluded from the analysis.

[0689] Subject characteristics: The baseline values of continuous and categorical covariates are summarized in Table 28. The descriptive statistics of the ADA samples are presented in Table 29.

[0690] Table 28: Summary of continuous and categorical covariates

[0691]

[0692]

[0693] Note: Subjects with at least one positive ADA sample were considered to have a positive ADA status.

[0694] Abbreviations: ADA = anti-drug antibody; BMI = body mass index; max = maximum; min = minimum; N = number of subjects with available information; N = number of subjects in a specific category;

[0695] SD = standard deviation

[0696] Table 29: Summary of ADA samples

[0697]

[0698] Abbreviations: ADA = anti-drug antibody; N = number of subjects with available information

[0699] PPK analysis: The arithmetic mean (+ standard deviation) serum drug concentration-time profiles of the study population are shown in Figure 20A and 20B . The shape of the elimination phase in the semi-logarithmic plot can indicate two-compartment kinetics, while the XTMAB-16 concentration-versus-time profile can indicate dose-proportional exposure upon dose increase.

[0700] Model development and quantification: A two-compartment model was used as the initial structural model. Inter-individual variability (IIV) was evaluated for all parameters. Due to the lack of improvement in IIV in the inter-compartment clearance (Q), only the improvement in IIV in clearance (CL), central volume of distribution (V), and peripheral volume of distribution (V2) was supported and retained in the model. A combined additive error and proportional error model was chosen to describe the residual error. Body weight and subject-level ADA were found to affect the PK of XTMAB-16. The parameter estimates of the final PPK model are presented in Table 30. Based on the final model, CL was 0.009 L / h, the volume of distribution at steady state was 3.02 L, and the estimated half-life for a typical individual (where weight was 72.4 kg) was 243.3 h (194.0 h in the case of ADA-positive subjects). The parameters of the final XTMAB-16 PPK model were estimated with good precision; the percentage of relative standard error for PK parameters and random effects was < 23%.

[0701] Table 30: Final model parameter estimates

[0702]

[0703] The final model resolved the residual covariate trends on body weight and sex for the central volume of distribution; however, there remained a small residual trend for the peripheral volume in terms of sex. However, adding the covariate on sex did not result in a significant decrease in the objective function value (Δ -2LL = 5.01), and the sex covariate was not included in the final model. Due to the small sample size, the effects of race and ethnicity were not strongly estimated.

[0704] The goodness-of-fit plot indicates reasonable fit of the data ( Figures 21A - 21F ). The predicted concentrations for individuals and the population showed good correlation with the observed data ( Figures 21A - 21D ). The CWRES were evenly distributed around 0 and did not show a trend over time or by concentration change, indicating lack of bias in the model (Figure 21E and 21F )。

[0705] pcVPC was performed to quantify the model, thereby simulating 1000 replicates of the observed subjects. The pcVPC curve (linear-log) of the final XTMAB-16PK model concentration is presented in Figure 22 . The final PPK model was able to predict the observed median and the 5th and 95th percentiles of the observed XTMAB-16 concentration with good accuracy. Model evaluation by pcVPC indicated that the model had appropriate predictive performance, and the model could capture both the central tendency and variability of the observed concentrations.

[0706] Simulation: Simulated XMTAB-16 serum and lung exposures increased with increasing dose. For a given mg / kg dose, compared with the Q4W dosing regimen, the Q2W dosing regimen produced higher trough concentrations (C 谷 ) and mean concentrations (C 评级 ) (Tables 31 and 32). The PK parameters of the simulated serum XTMAB-16 concentration after a single administration are shown in Table 33.

[0707] Table 31: Simulated XTMAB-16 exposure in serum at steady state (ADA negative)

[0708]

[0709] Table 32: Simulated XTMAB-16 exposure in lung at lung steady state (ADA negative)

[0710]

[0711] After considering the expected biodistribution of the mAb from plasma to lung tissue, simulations in adult subjects indicated that 4 mg / kg Q2W would largely achieve C 谷 values in the lung that exceeded the IC50s that exhibited reduced granuloma growth in an in vitro sarcoidosis model (granuloma area fraction IC50 = 5.197 μg / mL; IL-1μ IC50 = 3.465 μg / mL); while doses of 2 mg / kg Q2W or Q4W or 4 mg / kg Q4W would fall below the in vitro IC50s ( Figure 23 ). The predicted lung exposure for the C 平均值 was similar to the granuloma IC50 for the 4 mg / kg Q2W regimen, with the remaining regimens falling below the IC50. Therefore, it is predicted that the proposed clinical dose regimens will provide a range of XTMAB-16PK lung exposures in the steep portion of the exposure-response curve.

[0712] Table 33: Pharmacokinetic parameters of simulated serum XTMAB-16 concentration after single administration (ADA negative)

[0713]

[0714] The simulated serum XTMAB-16 concentration setting of Ceoi relative to the 2-hour nominal time.

[0715] Abbreviations: ADA = anti-drug antibody; AUC infinity = area under the concentration-time curve extrapolated from time 0 to infinity; Ceoi = concentration at the end of infusion; CL = clearance; Cmax = maximum concentration; C trough = trough concentration; MRT infinity = mean residence time; Q2W = every 2 weeks; Q4W = every 4 weeks; T1 / 2 = half-life; Tmax = time to reach maximum concentration; Vss = volume of distribution at steady state; Vz = apparent volume of distribution during the terminal phase

[0716] Example 9: Evaluation of XTMAB-16 in subjects with pulmonary sarcoidosis

[0717] This example will be conducted as a multiple ascending dose (MAD) / proof of concept (POC) study of XTMAB-16 in subjects with sarcoidosis with or without extrapulmonary involvement (hereinafter referred to as the Phase 2 project). This study consists of two parts: Part A will be a randomized, double-blind, placebo-controlled multiple-dose escalation study, and Part B will be a randomized, double-blind, placebo-controlled POC study ( Figure 24 ).

[0718] The objective of Part A is to evaluate the safety and tolerability of the MAD of XTMAB-16 and to determine the recommended Phase 2 dose and frequency of XTMAB-16 to be administered to subjects with sarcoidosis with or without extrapulmonary manifestations. The objectives and endpoints of Part A are outlined in Tables 34 - 36.

[0719] The objective of Part B is to confirm the preliminary efficacy of XTMAB-16, as measured by the ability to reduce the use of background oral corticosteroids in subjects with sarcoidosis with or without extrapulmonary manifestations. The objectives and endpoints of Part B are outlined in Table 37.

[0720] The rationale for this study is to conduct dose range finding to identify the recommended Phase 2 dose (RP2D) for Part B and to characterize various clinical and biomarker outcome measures to demonstrate the efficacy and POC of XTMAB-16 in this subject population.

[0721] Table 34: Primary objectives and endpoints of Part A (MAD)

[0722]

[0723] Table 35: A Part (MAD) Secondary Objectives and Endpoints

[0724]

[0725]

[0726] Table 36: A Part (MAD) Exploratory Objectives and Endpoints

[0727]

[0728] Abbreviations: 6MWD = 6-minute walk distance; ACE = angiotensin-converting enzyme; ADA = anti-drug antibody; AE = adverse event; AESI = adverse event of special interest; AUC = area under the curve; AUEC = area under the effect curve; C 平均值 = mean concentration; C 最大值 = maximum observed concentration; CRP = C-reactive protein; C 谷 = trough concentration; DLT = dose-limiting toxicity; E 最大值 , observed = maximum observed effect; EOI = end of infusion; FAS = Fatigue Assessment Scale; FVC = forced vital capacity; HRCT = high-resolution computed tomography; IL-1b = interleukin 1b; IL-6 = interleukin-6; KSQ = King's Sarcoidosis Questionnaire; LCQ = Leicester Cough Questionnaire; MAD = multiple ascending dose; mg = milligram; mMRC = modified Medical Research Council; nAb = neutralizing antibody; PD = pharmacodynamics; PGA = patient global assessment;

[0729] PK = pharmacokinetics; QoL = quality of life; SAE = serious adverse event; SF-36 = Short Form-36; sIL 2R = soluble interleukin-2 receptor; sTNFα = soluble tumor necrosis factor α; STQ = Steroid Toxicity Questionnaire; tE 最大值， observed = time to reach maximum observed effect.

[0730] Table 37: B Part (POC) Primary Objectives, Secondary Objectives, Exploratory Objectives and Endpoints

[0731]

[0732]

[0733] Abbreviations: 6MWD = 6-minute walk distance; ACE = angiotensin-converting enzyme; ADA = anti-drug antibody; AE = adverse event; AUC = area under the concentration-time curve; C 平均值 = mean concentration;

[0734] C 最大值= Maximum Observed Concentration; CRP = C-Reactive Protein; C 谷 = Trough Concentration; FAS = Fatigue Assessment Scale; FVC = Forced Vital Capacity; HRCT = High-Resolution Computed Tomography;

[0735] IL-1b = Interleukin-1b; IL-6 = Interleukin-6; KSQ = King's Sarcoidosis Questionnaire; LCQ = Leicester Cough Questionnaire; mg = milligram; mMRC = Modified Medical Research Council; nAb = Neutralizing Antibody; PD = Pharmacodynamics; PGA = Patient Global Assessment; PK = Pharmacokinetics; POC = Proof of Concept;

[0736] SAE = Serious Adverse Event; SF-36 = Short Form-36; sIL 2R = Soluble Interleukin-2 Receptor; sTNFα = Soluble Tumor Necrosis Factor α; STQ = Steroid Toxicity Questionnaire.

[0737] Overall Design

[0738] Example 9 includes a Multiple Ascending Dose (MAD) / POC study of XTMAB-16 to evaluate the safety, tolerability, immunogenicity, pharmacokinetics, and preliminary efficacy of XTMAB-16 in subjects with pulmonary sarcoidosis with or without extrapulmonary involvement. Part A will consist of a screening period of up to 42 days and a 12-week randomized, double-blind, placebo-controlled treatment period of the MAD protocol ( Figure 25 ), in which subjects will be randomized to XTMAB-16 or placebo at a 3:1 ratio per cohort (6 active, 2 placebo). The Recommended Phase 2 Dose (RP2D) will be determined based on the results of Part A, and Part B will commence as a double-blind, placebo-controlled POC study.

[0739] Subjects who complete Part A will maintain their assigned dosing regimen in a rollover long-term open-label safety study until the Data Safety Monitoring Committee (DSMC) has evaluated all doses. After the RP2D for Part B has been selected, subjects who are in treatment in the rollover study will transition to the RP2D.

[0740] Part B will consist of a double-blind, placebo-controlled POC study using the RP2D determined in Part A with a 1:1 XTMAB-16 to placebo assignment ratio for 24 weeks ( Figure 26 ). At the completion of Part B, all subjects who have completed up to 24 weeks of treatment and evaluation will be offered the opportunity to roll over into an ongoing long-term open-label safety study.

[0741] Number of Subjects

[0742] Up to 32 subjects will be enrolled in Part A to receive XTMAB-16 or placebo in a MAD manner in planned cohorts for 12 weeks: 2 mg / kg every four weeks (Q4W) for 12 weeks (n = 6 active, 2 placebo), 4 mg / kg Q4W for 12 weeks (n = 6 active, 2 placebo), 2 mg / kg every two weeks (Q2W) for 12 weeks (n = 6 active, 2 placebo), 4 mg / kg Q2W for 12 weeks (n = 6 active, 2 placebo).

[0743] Approximately 62 subjects will be enrolled in Part B to receive XTMAB-16 or placebo (1:1) in a randomized, double-blind, placebo-controlled manner over 24 weeks. The baseline dose of prednisone or equivalent will be used as a stratification factor during randomization.

[0744] All subjects who complete participation in the study in Part A or Part B will be offered the opportunity to roll over to an open-label long-term safety study.

[0745] Summary of major eligibility criteria: Inclusion criteria

[0746] 1. Subjects are between 18 and 80 years of age (inclusive).

[0747] 2. Weigh between 45 kg and 160 kg (99 lbs to 353 lbs) at screening.

[0748] 3. Are diagnosed with pulmonary sarcoidosis (at least 6 months prior to screening) using the 2020 American Thoracic Society (ATS) Clinical Practice Guidelines, European Respiratory Society (ERS), or World Association of Sarcoidosis and Other Granulomatous Disorders (WASOG) criteria, including excluding clinical and radiological manifestations compatible with other causes of granulomatous disease (cutaneous and ocular involvement is permitted).

[0749] 4. Modified Medical Research Council (mMRC) dyspnea scale ≥ 1.

[0750] 5. Are receiving treatment with oral prednisone or equivalent at 7.5 mg / day to 25 mg / day during the screening period and at the discretion of the investigator to be able to conduct the protocol-specific corticosteroid taper protocol.

[0751] 6. Have received treatment with methotrexate, azathioprine, mycophenolate mofetil, leflunomide, chloroquine, or hydroxychloroquine for at least 3 months before screening and have been at a stable dose for 4 weeks before screening. At the discretion of the investigator, every effort should be made to maintain a stable background therapy at the screening dose throughout the intervention period.

[0752] 7. Only for Part A: Willing to avoid grapefruit or grapefruit juice [pomelos, exotic citrus fruits, or grapefruit hybrids] from the screening visit until after the final dose administration.

[0753] 8. Negative for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by polymerase chain reaction (PCR) test or rapid antigen test at screening.

[0754] 9. Able to provide written informed consent.

[0755] 10. In the opinion of the investigator, the subject is able to understand and comply with the protocol requirements.

[0756] Summary of major eligibility criteria: Exclusion criteria

[0757] 1. Pregnant women, or women who are breastfeeding or plan to become pregnant during the study.

[0758] 2. Only for Part A: Subjects > 65 years of age.

[0759] 3. Only for Part A: Having a known potentially significant fibrotic disease and / or active inflammation only in the hilar region, as shown by high-resolution computed tomography (HRCT) and confirmed by a central reader. Subjects with current active inflammation in the hilar region and concurrent inflammation outside the hilar region may be included. For subjects with disease onset < 2 years, a historical computed tomography (CT) confirmed by central reading performed within 6 months before screening is acceptable. For subjects with disease onset > 2 years and no CT performed within 6 months before screening, a CT will be performed at screening.

[0760] Note: For all subjects, regardless of the time of disease onset, when submitting historical HRCT for diagnostic confirmation, the HRCT must have been performed within 6 months of screening. In the case of having had their last HRCT > 6 months before screening, then they will require an HRCT performed during screening for diagnostic confirmation.

[0761] Note: A significant fibrotic disease is defined as > 20% fibrosis on HRCT.

[0762] 4. Only for Part A: Any previous tumor necrosis factor α (TNFα) inhibitor therapy.

[0763] 5. Clinically significant extrapulmonary sarcoidosis requiring systemic therapy, as determined by the investigator.

[0764] 6. Part B only: Any therapy with an anti-TNFα monoclonal antibody (e.g., infliximab, adalimumab, golimumab, and biosimilars thereof) within 6 months.

[0765] 7. Baseline predicted FVC percentage < 50%.

[0766] 8. Prior treatment with rituximab or depot corticotropin injection within the previous 12 months.

[0767] 9. Clinically significant central nervous system (CNS) sarcoidosis requiring therapy, except for a history of isolated seventh cranial nerve palsy or evidence of a demyelinating neurological disease.

[0768] 10. Advanced congestive heart failure (New York Heart Association [NYHA] class 3 or 4).

[0769] 11. Current disease manifestations consistent with Löfgren's syndrome (i.e., presence of the triad of erythema nodosum, bilateral hilar lymphadenopathy on chest X-ray, and joint pain).

[0770] 12. Clinically significant pulmonary hypertension requiring therapy.

[0771] Note: Clinically significant pulmonary hypertension requiring treatment will be defined as treatment with prostanoids, phosphodiesterase 5 inhibitors, and endothelin receptor antagonists.

[0772] 13. Known hypersensitivity to any component of the formulation of XTMAB-16.

[0773] 14. During the study participation, planned liver or messenger ribonucleic acid (mRNA) vaccination or vaccination with live or mRNA vaccines within 2 weeks before Day 1.

[0774] 15. Active or latent tuberculosis (TB) demonstrated by interferon-γ release assay (IGRA) or invasive fungal infection at screening.

[0775] 16. Known positive history of malignancy within the last 2 years, except for melanoma skin cancer, including cervical carcinoma in situ that has been completely cured by radical surgery.

[0776] 17. Positive test results for a known history of hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) antibody, coronavirus disease (COVID-19), TB, or human immunodeficiency virus (HIV) infection at screening.

[0777] 18. Women who are sexually active with an unsterilized male partner of reproductive potential and are unwilling to adhere to adequate birth control measures starting from signing the informed consent form, throughout the duration of the study, and for 90 days after 5 half-lives from the last administration of the study drug.

[0778] 19. Unsterilized male subjects who are sexually active with a fertile female partner and are unwilling to use adequate contraception starting from signing the informed consent form, throughout the duration of the study, and for 90 days after 5 half-lives from the last administration of the study drug.

[0779] 20. Clinically significant liver or kidney disease, including diabetes that is uncontrolled at the discretion of the investigator.

[0780] 21. Any previous severe reaction to any type of biologic or human blood product (such as albumin, immunoglobulin G (IgG), etc.).

[0781] 22. Concurrent emphysema.

[0782] 23. Known hypercalcemia due to non-sarcoidosis conditions, such as untreated hyperparathyroidism, at the discretion of the investigator.

[0783] 24. Electrocardiogram (ECG) abnormalities: ventricular arrhythmias (non-sustained ventricular tachycardia (VT), multifocal or frequent ventricular premature beats, bundle branch block, axis deviation, or abnormal Q waves). In cases where the QTcF (QT interval corrected by Fredericia) interval > 450 milliseconds (male) or > 480 milliseconds (female; subjects with bundle branch block) or the PR interval is outside the range of 120 milliseconds to 220 milliseconds, the assessment can be repeated once at screening or baseline for eligibility determination.

[0784] 25. Donating or losing 450 mL or more of their blood volume (including plasmapheresis) or receiving any blood product within 90 days before dosing.

[0785] 26. Known uncontrolled hypertension.

[0786] Note: Despite undergoing anti-hypertensive therapy within 3 months of randomization, uncontrolled hypertension is still defined as a blood pressure ≥ 160 / 100 mmHg.

[0787] 27. Clinical signs and symptoms consistent with COVID-19, such as fever, dry cough, shortness of breath, sore throat, fatigue, new olfactory or taste disorders, or infection confirmed by appropriate laboratory tests, occurring within the last 4 weeks before screening.

[0788] 28. In the opinion of the researchers, it is not possible to tolerate the tapering of corticosteroids.

[0789] 29. Conduct parallel systemic steroid use for non-sarcoid conditions.

[0790] 30. Concurrent known autoimmune diseases that require treatment.

[0791] 31. Participate in another clinical trial of the study drug within 3 months (for small molecules) / 6 months (for biologics) of the drug or 5 half-lives (if known), whichever is longer.

[0792] 32. Any condition that requires hospitalization within 3 months prior to Day 1 or may require hospitalization during the study.

[0793] 33. Clinically significant abnormalities in the screening physical examination, medical history, vital signs, ECG, or clinical laboratory tests, which are known not to be due to concurrent sarcoidosis and, in the opinion of the researchers and medical monitors, should exclude the subject from participating in the clinical study.

[0794] Lifestyle Considerations

[0795] During the course of this study, subjects will be asked to:

[0796] 1. Avoid consuming red wine and Seville oranges from the screening visit until after the final dose. Additionally, subjects should also avoid consuming grapefruit or grapefruit juice (pomelos, exotic citrus fruits, or grapefruit hybrids) and any medications or diets that may interact with drug-metabolizing enzyme activity from the screening visit to after the final dose. This is necessary because grapefruit enhances cortisol availability and interferes with the tapering of corticosteroids that is essential for the study endpoint.

[0797] 2. Minimize interaction with household contacts who may be immunocompromised.

[0798] 3. Be vaccinated against COVID-19 at least 2 weeks prior to participation, which is defined as the initial 2-dose regimen of the Moderna vaccine or the Pfizer vaccine or a single J&J vaccine.

[0799] Screening Failure

[0800] Screening failure is defined as a subject who consented to participate in the clinical trial but was subsequently not randomly assigned or allocated to the study intervention or did not enter the study.

[0801] The minimum set of failure information needs to be screened to ensure transparency in reporting failed screenings of subjects, to meet the publication requirements of the Consolidated Standards of Reporting Trials (CONSORT), and to respond to regulatory inquiries. The minimum information includes demographics, details of screening failures, and eligibility criteria.

[0802] Individuals who do not meet the criteria for participation in this trial (screening failure) may be rescreened once at the discretion of the investigator after consultation with the medical monitor. The same subject number as that assigned to the subject at initial screening should be assigned to the rescreened subject.

[0803] Treatment failure

[0804] A subject will be considered to have treatment failure if the subject experiences any of the following:

[0805] 1. Addition of background second-line antimetabolite therapy. Note that the dose of the antimetabolite therapy can be adjusted to avoid toxicity.

[0806] 2. A steroid salvage period greater than 1.

[0807] 3. Breakthrough extrapulmonary disease requiring systemic treatment (local and ophthalmic therapies for concurrent skin and eye diseases are permitted).

[0808] 4. Failure to achieve the expected tapered dose of 5 mg by week 12.

[0809] 5. Failure to maintain a tapered or lower dose (Part B) by week 24.

[0810] 6. Failure to taper steroids to the target maintenance dose by week 12 or week 24 after the salvage period.

[0811] 7. A steroid salvage period requiring treatment longer than 2 weeks.

[0812] 8. Permanent drug discontinuation due to at least two grade 3 or one grade 4 toxicities considered by the investigator to be related to XTMAB-16 treatment (for analysis purposes, discontinuation of the study drug will be considered treatment failure).

[0813] Study intervention and concomitant therapies

[0814] The study intervention will be conducted as outlined in Table 38:

[0815] Table 38: Study treatment formulation and preparation

[0816]

[0817] Abbreviations: IV = intravenous; mg = milligram; SWFI = Sterile Water for Injection. Source: Pharmacy Handbook

[0818] Administration and Dosage: Part A Dose Escalation:

[0819] During Part A, multiple doses and frequencies will be explored in an escalating manner ( Figure 10 and 13). To identify the recommended Phase 2 dose for Part B, dose level groups will be enrolled sequentially and in parallel by dose density as described below, and the DSMC will continuously review the cumulative safety data.

[0820] Part A will consist of 32 subjects in 4 cohorts, each cohort consisting of 8 subjects, who will be randomly assigned in a 3:1 ratio (6 active, 2 placebo). The first cohort in the 2 mg / kg Q4W dose group will initiate subject enrollment initially. After completing the safety assessment of at least 4 subjects who have completed 3 administrations in this dose group, the dose is considered safe to continue dosing in parallel in the second and third cohorts at 4 mg / kg Q4W and 2 mg / kg Q2W, as the dose densities of these regimens are considered equal.

[0821] After completing the safety assessment of at least 4 subjects in both the 4 mg / kg Q4W dose group and the 2 mg / kg Q2W dose group (3 administrations completed in the Q4W cohort and 6 administrations completed in the Q2W cohort), the dose will be considered safe to continue dosing in the fourth cohort at 4 mg / kg Q2W.

[0822] The DSMC may make an independent determination of the safety of the 4 mg / kg Q4W and 2 mg / kg Q2W dose groups in the second and third cohorts to determine if it is safe to proceed to the next dose group.

[0823] The escalation protocol will be carried out according to the protocol shown in Figure 27 .

[0824] Unless the minimum follow-up is not reached in a cohort, enrollment will not be paused between cohorts, and enrollment will continue during regular safety reviews. After the regular safety data review, if any safety signals or DLTs are identified in the judgment of the DSMC, dose escalation and enrollment in all dose groups may be paused for a complete safety evaluation. The criteria for DLT are as follows: Any prolonged Grade 2 (defined as lasting more than 3 weeks at Grade 2 (or higher)) or any Grade 3 or higher adverse event considered related to the study drug and not attributable to deterioration of concurrent illness.

[0825] The stopping criteria for safety signals that may require DSMC to call for additional safety reviews during the dose escalation period are as follows:

[0826] 1. Any SAE evaluated as related to the investigational drug

[0827] 2. Any signs of serious infection such as TB, bacterial sepsis or invasive fungal infection or lymphoma diagnosis

[0828] 3. Grade 3 hypersensitivity or allergy

[0829] Completion of the extended DSMC safety review may enable resumption of enrollment, continuation of dose escalation, expansion of the current dose level cohort, or termination of enrollment. Dose escalation will continue in this manner until the highest planned dose level is complete or the DSMC decides to abort enrollment due to safety concerns. If needed, unplanned PK sample, ECG, or clinical laboratory data analysis may be conducted in the event of a safety signal of interest to support safety assessment.

[0830] Dose selection: Part B (POC)

[0831] After the DSMC completes and reviews the data from Part A of this study, the dose levels and frequencies for the POC part of Part B of this study will be determined. Dose determination will be guided by the observed meaningful relevant PK parameters (C 谷 , AUC) for each cohort, the relationship between the observed PK and in vitro measures of pharmacological activity, the changes in observed clinically meaningful biomarkers (ACE, sIL2R, CRP, IL-1b), the observed clinical benefits (ability to reduce background oral corticosteroid use, QoL, and changes in lung endpoints), the observed safety signals, and DLT. If needed, exploratory efficacy and / or PK data analysis may be conducted to select the recommended Phase 2 dose for Part B.

[0832] Oral corticosteroid tapering regimen: Part A (MAD)

[0833] During Part A of the study, two weeks after the first dose is administered and prior to the second administration to subjects assigned to the Q2W dosing regimen, subjects will begin tapering corticosteroids from their starting dose of prednisone to a target dose of 5 mg / day or an equivalent form or less to be completed by week 12 or earlier. Corticosteroids will be tapered according to the starting dose as shown in Table 39. The corticosteroid dose for subjects may be further titrated to below 5 mg / day or discontinued if feasible.

[0834] In cases where the absolute deterioration rate of the subject's FVC% is >5% (at two visits spaced at least 7 days apart), the oral corticosteroid dose of the subject can be increased at the discretion of the investigator during a two-week rescue therapy period. A change in score of the King's Sarcoidosis Questionnaire Lung Module (KSQ Lung) of a decrease of ≥4 points and / or a decrease in the Leicester Cough Questionnaire (LCQ) score of ≥1 point, or the investigator determines that a rescue dose of corticosteroid is required due to worsening cough, shortness of breath, exercise intolerance, or symptom exacerbation will confirm symptom flare. The subject should be evaluated at the clinic when possible or via telemedicine.

[0835] In cases where there is no clinical indication for an increase in steroid dose, steroid rescue therapy is not mandatory in cases of worsening cough or dyspnea or change in FVC. Subjects who need to increase their steroid dose at any time during the study should continue to receive the study drug and be followed until the end of the study. The subject will record the steroid dose in a diary, which will be reviewed at the planned telephone contact and site visit.

[0836] For subjects who require corticosteroid rescue therapy between baseline and week 12, it can be continued to be gradually reduced at the discretion of the investigator after the acute event has subsided. The tapering of corticosteroids must be completed by week 12, whether it is rescue therapy or not, and will be considered a treatment success. Failure to taper after rescue therapy will constitute treatment failure. The subject will be allowed to have at most one course of corticosteroid rescue therapy between baseline and week 12 to be considered responsive to treatment. However, additional corticosteroid rescue therapy can be given to the subject if the investigator determines it is necessary.

[0837] Table 39: Corticosteroid tapering protocol by starting dose: Part A (MAD)

[0838]

[0839] Abbreviations: MAD = multiple ascending doses; mg = milligram.

[0840] The doses reflected in Table A are prednisone doses.

[0841] Oral corticosteroid tapering protocol: Part B (POC)

[0842] During Part B of the study, two weeks after completion of the first dosing and prior to the second administration, subjects will begin to taper corticosteroids from their starting dose of prednisone to a target dose of 5 mg / day or equivalent or less, to be completed by week 12 and maintained through week 24 (Table 40). Corticosteroids will be tapered based on the starting dose and at the discretion of the investigator. At the discretion of the investigator, the subject's corticosteroid dose may be further titrated below 5 mg / day or discontinued if feasible.

[0843] In the event of an absolute decline rate of >5% in the subject's FVC% (2 visits spaced at least 7 days apart), the subject's corticosteroid dose may be increased at the discretion of the investigator during a two-week rescue therapy period. A change in the KSQ lung score of a decrease of ≥4 points and / or a decrease in the LCQ score of ≥1 point, or the investigator determines that a rescue dose of corticosteroids is required due to worsening cough, shortness of breath, exercise intolerance, or worsening symptoms will confirm symptom flare. Subjects should be evaluated in the clinic or via telemedicine.

[0844] In the absence of a clinical indication for an increase in steroid dose, steroid rescue therapy is not mandatory in the event of worsening cough or dyspnea or a change in FVC. Subjects who require an increase in their steroid dose at any time during the study should continue to receive the blinded study product and be followed until the end of the study. Subjects will record their steroid dose in a diary, which will be reviewed at the time of the relevant scheduled phone contact and site visit.

[0845] For subjects who require corticosteroid rescue therapy between baseline and week 12, tapering may continue at the discretion of the investigator after the acute event has resolved. Tapering of corticosteroids must be completed by week 12 and maintained through week 24, whether or not it is rescue therapy, and will be considered a treatment success. Failure to taper after rescue therapy will constitute treatment failure. Subjects will be allowed up to one corticosteroid rescue therapy to be considered responsive to treatment; however, additional corticosteroid rescue therapy may be given to the subject if determined necessary.

[0846] Table 40: Corticosteroid Tapering Protocol by Starting Dose: Part B (POC)

[0847]

[0848] Abbreviations: mg = milligram; POC = proof of concept.

[0849] The doses reflected in Table A are prednisone doses.

[0850] Prepare XTMAB-16

[0851] The manufacturing process of XTMAB-16 (drug product) for injection includes steps of compounding, filtration, filling, semi-stoppering, freeze-drying, full-stoppering and sealing. The drug product is sterilized by filtration before aseptic filling. No impurities are introduced during the manufacturing of the drug product.

[0852] The freeze-dried drug product XTMAB-16 is provided in the form of a sterile white to off-white freeze-dried powder or a freeze-dried cake (100 mg of XTMAB-16 per vial) in 20 mL vials for reconstitution with 10 mL of sterile water for injection (SWFI) and intravenous infusion. The concentration of the reconstituted solution is 10 mg / mL. The pH is between 5.8 and 6.2. The reconstituted solution is diluted with 0.9% sodium chloride injection of the United States Pharmacopeia (USP) to a range between 0.4 mg / mL and 4 mg / mL for infusion.

[0853] The matching placebo is compounded in the same way as the drug product, except that it does not contain XTMAB-16.

[0854] Randomization and Blinding

[0855] In both Part A (MAD) and Part B (POC) of this study, subjects will be randomly assigned to treatment (XTMAB-16 or placebo), and the treatment assignment will be double-blind; during the study duration, all researchers will be unaware of the subjects' treatment assignments. The infusion solutions containing the active drug and the placebo will be indistinguishable in appearance. If possible, all study evaluations and causality will be conducted before unblinding.

[0856] Previous Therapies and Concomitant Therapies

[0857] The previous medication history up to one year before screening will be collected. Concomitant medications are permitted during the study, except as specified in the inclusion or exclusion criteria or study procedures. The name, indication, dose and duration of use of the medications will be recorded on the eCRF. In the case of starting a new concomitant therapy due to an adverse event, the adverse event must be recorded on the eCRF.

[0858] The dose of concomitant corticosteroids will be self-recorded in the subject's diary and verified by the site personnel at each in-person or telephone visit.

[0859] Subjects will receive concomitant treatment with methotrexate, azathioprine, mycophenolate mofetil, leflunomide, chloroquine or hydroxychloroquine at a stable dose for at least 3 months before screening and for 4 weeks before screening. Every effort should be made to maintain the stable background therapy at the screening dose throughout the intervention period.

[0860] During the screening period, the subjects will also receive treatment with oral prednisone or its equivalent form at 7.5 mg / day to 25 mg / day, which will be withdrawn during the study period.

[0861] Study Assessments and Procedures: Efficacy Assessments

[0862] The planned time points for all efficacy assessments are provided in the SoA (Tables 44 - 51).

[0863] Study Assessments and Procedures: Quality of Life Measures

[0864] All quality of life assessments should be conducted prior to other study procedures at each scheduled visit and should be completed in the same order at each scheduled visit. The order of quality of life measurements will be outlined in the electronic clinical outcomes assessment (eCOA) site manual.

[0865] Study Assessments and Procedures: King's Sarcoidosis Questionnaire

[0866] The King's Sarcoidosis Questionnaire (KSQ) is a 29 - item online questionnaire to be completed by sarcoidosis subjects. The KSQ is divided into 5 sections: general health status, lungs, medications, skin, and eyes. Results are given as a number between 1 and 100, with higher numbers indicating better health status. The questionnaire takes approximately 10 minutes to complete. The minimal clinically important difference (MCID) for the KSQ in sarcoidosis is still unclear.

[0867] Study Assessments and Procedures: Leicester Cough Questionnaire

[0868] The Leicester Cough Questionnaire (LCQ) is a 19 - item self - completed quality of life measure in response to variable chronic cough. The items on the scale are divided into 3 domains: physical, psychological, and social. The items are scored on a 7 - point Likert scale. The total score (range 3 - 21) is also calculated by adding the domain scores, with higher scores indicating better quality of life. The questionnaire takes 5 to 10 minutes to complete. The MCID for chronic cough is 1.3.

[0869] Study Assessments and Procedures: Short Form - 36

[0870] The Short Form - 36 (SF - 36) is a 36 - item subject - reported health survey. The SF - 36 includes a multi - item scale that assesses eight health concepts: 1) limitations in physical activity due to health problems; 2) limitations in social activities due to physical or emotional problems; 3) limitations in usual role activities due to physical health problems; 4) bodily pain; 5) general mental health (psychological distress and well - being); 6) limitations in usual role activities due to emotional problems; 7) vitality (energy and fatigue); and 8) general health perception.

[0871] Research Assessments and Procedures: Fatigue Assessment Scale

[0872] The Fatigue Assessment Scale (FAS) contains 10 specific fatigue questions that have been validated in sarcoidosis subjects. For each question, one of 5 answer categories can be selected from never to always: 1 = never; 2 = sometimes (about once a month or less); 3 = regularly (about several times a month); 4 = often (about once a week); 5 = always (about daily). An answer must be given for each question, even if the person currently has no complaints. The scores for questions 4 and 10 should be recorded (1 = 5, 2 = 4, 3 = 3, 4 = 2, 5 = 1). Subsequently, the total FAS score can be calculated by adding up the scores for all questions (the recoded scores for questions 4 and 10). The sum of questions 3 and 6 to 9 represents mental fatigue, and the sum of questions 1, 2, 4, 5, and 10 represents physical fatigue. The minimum score is 10, and the maximum score is 50. A score of ≥22 is considered to indicate substantial fatigue. A change of 4 points in the FAS score is considered to be the MCID. The FAS takes about 5 to 10 minutes to complete.

[0873] Research Assessments and Procedures: Patient Global Assessment

[0874] The Patient Global Assessment (PGA) is a subject-reported outcome tool that is used to measure the well-being of the subject and is scored using a Visual Analogue Scale (VAS). In randomized controlled trials, the MCID in sarcoidosis remains unclear. The previously reported MCID for PGA in sarcoidosis was <7 cm, 7 cm to 10 cm.

[0875] Research Assessments and Procedures: Steroid Toxicity Questionnaire

[0876] Glucocorticoids (GC) are associated with clinically significant toxicity in sarcoidosis subjects, related to both the cumulative dose and the duration of treatment. The Steroid Toxicity Questionnaire (STQ) is a simple 12-question tool that uses a 4-point scale to characterize the toxicity associated with steroid use. The MCID in sarcoidosis remains unclear.

[0877] Research Assessments and Procedures: Modified Medical Research Council Dyspnea Scale

[0878] The Modified Medical Research Council (mMRC) Dyspnea Scale is a five-level rating scale based on the subject's perception of dyspnea during daily activities, ranging from no dyspnea (grade 0) to very severe dyspnea (grade 4). The score is the number that best matches the subject's activity level.

[0879] Research Assessments and Procedures: Pulmonary Function Tests

[0880] Pulmonary function tests (PFTs), including forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), and FEV1 / FVC ratio, will be determined by spirometry at the time points specified in the SoA (Tables 44 - 51). Spirometry tests will be performed according to ATS / ERS guidelines. A spirometer (SpiroSphere) and related equipment will be provided to each study site. The same spirometry equipment must be used for all evaluations of a given subject. All subjects visited during the trial will be evaluated. Whenever possible, the same technician should perform all maneuvers on an individual subject. All staff performing spirometry tests must be appropriately trained and pass two proficiency tests, which must be documented. Spirometry should ideally be performed on subjects in the seated position for at least 2 minutes with a tight seal around the mouth and a nose clip worn or the nostrils manually occluded; the subject should remain seated and the head slightly elevated throughout the test procedure. If testing a subject in another position, this must be documented in the report. It is recommended to perform the test at least 1 hour after a light meal, avoid smoking and / or e-cigarettes and / or using a hookah within 1 hour before the test, avoid consuming intoxicants within 8 hours before the test, avoid wearing clothing that significantly restricts full chest and abdominal expansion, and avoid strenuous exercise within 1 hour after the test. In cases of treatment with a bronchodilator, the subject should discontinue the medication prior to spirometry according to the guidelines listed in Table 47. Spirometry should be performed in the same manner at all evaluation time points.

[0881] Table 41: Guidelines for Discontinuation of Bronchodilator Medications

[0882] Bronchodilator drug Withdrawal time SABA (e.g., albuterol / salbutamol) 4 hours to 6 hours SAMA (e.g., ipratropium bromide) 12 hours LABA (e.g., formoterol or salmeterol) 24 hours Ultra-LABA (e.g., indacaterol, vilanterol or olodaterol) 36 hours LAMA (e.g., tiotropium bromide, umeclidinium, aclidinium or glycopyrronium) 36 hours to 48 hours

[0883] Abbreviations: LABA = long-acting β2-agonist; LAMA = long-acting muscarinic antagonist; SABA = short-acting β2-agonist; SAMA = short-acting muscarinic antagonist.

[0884] Study Evaluations and Procedures: Six-Minute Walk Distance

[0885] The six-minute walk distance (6MWD) will record the distance walked in six minutes, the oxygenation level will be recorded using a pulse oximeter, and the level of dyspnea will be recorded using a dyspnea scale of 0 - 10 (Borg score). The standardized protocol for the six-minute walk will be for all sites and must be performed in the same manner at all time points.

[0886] Study Evaluations and Procedures: High-Resolution Computed Tomography

[0887] High-resolution computed tomography (HRCT) is the preferred modality for evaluating the features of thoracic sarcoidosis. It accurately depicts the features of parenchymal nodules and lesions, their distribution, associated changes, and atypical features. It also helps guide appropriate therapy by differentiating active lesions from irreversible fibrosis.

[0888] Safety and Other Assessments

[0889] The planned time points for all safety and other assessments are provided in the SoA (Tables 44 - 51).

[0890] Safety and Other Assessments: Physical Examination

[0891] A complete physical examination will include an assessment of general appearance, skin; head and neck; pulmonary system, cardiovascular system, gastrointestinal system, external urogenital system (optional), lymphatic system, and musculoskeletal system; extremities; eyes (examination and vision control); nose; throat; and neurological status. A brief physical examination will include an assessment of general appearance; head, ears, eyes, nose, and throat; cardiovascular system; respiratory system; and gastrointestinal system (including abdomen). A targeted physical examination may be performed at any time at the discretion of the investigator and should be limited to clinically relevant systems (i.e., cardiovascular, respiratory, lymph nodes, liver).

[0892] Safety and Other Assessments: Height and Weight

[0893] Height (cm) will be measured only at screening. Weight will be measured in kilograms at each visit according to the SoA (Tables 44 - 51).

[0894] Safety and Other Assessments: Vital Signs

[0895] Oral, tympanic, or temporal temperature, respiratory rate, pulse, and systolic and diastolic blood pressure (BP) will be assessed. The method of temperature assessment chosen should be recorded and should be consistent throughout the study. In the case of extracting oral temperature (°C), the subject should not have consumed hot or cold beverages or smoked recently.

[0896] During infusion, systolic / diastolic blood pressure and pulse should be measured in the semi-supine / semi-recumbent position. For all other readings, blood pressure and pulse should be extracted after a 10 - 15 minute rest in the supine position. During infusion, vital signs should be collected according to the standard care intervals at the study site.

[0897] Respiratory rate (breaths / minute) should be extracted after at least a 5-minute rest; count the number of breaths for 30 seconds and multiply by 2. Respiratory rate will be extracted once during an ECG. During infusion, respiratory rate should be extracted in the semi-supine / semi-recumbent position.

[0898] Safety and Other Assessments: Electrocardiogram

[0899] The standard 12-lead ECG will be recorded when the subject has been resting in the supine position for at least 10 minutes. The ECG will be collected in triplicate at the time points mentioned in the SoA (Tables 44 - 51), with approximately 1 minute between each ECG; all other ECGs will be collected as single recordings. Additional ECGs will be made at the discretion of the investigator.

[0900] The investigator will be responsible for reviewing the ECG to assess whether it is within the reference limits and to determine the clinical significance of the results. The investigator has the authority to overrule the interpretation and intervals as measured by the ECG machine, if necessary. The respiratory rate will be extracted once during the ECG. The ECG will be evaluated to obtain the following metrics: P wave, QRS complex, U wave, QRS duration, QT interval, T wave, ST segment, RR interval, PR interval, and quantitative results.

[0901] Safety and Other Assessments: Clinical Laboratory Evaluations

[0902] Clinical laboratory evaluations (Tables 44 - 51) will be collected at the time points specified in the SoA under the appropriate conditions defined by the practice standards of the outpatient unit.

[0903] A central laboratory will be used to perform all clinical laboratory tests except for urine pregnancy tests, and the clinical laboratory tests will be evaluated by the site staff. For any safety concerns based on laboratory results, the investigator should take immediate action.

[0904] Record any clinically relevant changes that occur in the EDC system during the study. Laboratory reports must be filed together with the original documents. Clinically significant abnormal laboratory findings are abnormalities not related to underlying disease, unless the investigator determines that the subject's condition is more severe than expected.

[0905] All laboratory tests with values considered to be clinically significant during the study participation, including the subject's last EOS visit, should be repeated until the values return to normal or baseline, or until they are no longer considered clinically significant or no longer require follow-up according to the judgment of the investigator or medical monitor.

[0906] In the case where such values do not return to normal / baseline within a reasonable period of time as judged by the investigator, the cause should be identified and the sponsor notified.

[0907] Safety and Other Assessments: Computed Tomography

[0908] Images of computed tomography (CT) scans should be centrally read and evaluated by a central reader using a three-point Likert scale (worst to best).

[0909] Pharmacokinetics

[0910] Whole blood samples will be collected to measure the serum concentration of XTMAB-16. Instructions for the collection and handling of biological samples will be provided by the sponsor in a separate laboratory manual. The actual date and time (in 24-hour format) of each sample will be recorded in the eCRF. The samples will be used to evaluate the serum concentration and PK of XTMAB-16. Each serum sample will be divided into 2 aliquots (1 for primary and 1 for backup).

[0911] The concentration of XTMAB-16 will be measured using a validated assay method under the supervision of the sponsor or a qualified designee. The analytical method used to measure the concentration of XTMAB-16 will be described in a separate bioanalytical report. Only samples that are within the stability window of the sample will be analyzed.

[0912] Although PK samples must be collected from subjects assigned to the placebo group to maintain the blindness of the treatment assignment, the PK assay results for these subjects are not required for the safety guidelines or proper interpretation of this trial and therefore most samples will not be analyzed. The person responsible for performing the PK analysis will not be blinded to the treatment assignment of the subjects to identify the appropriate PK samples to be analyzed. Samples from subjects assigned to placebo may be analyzed upon request (i.e., to evaluate possible dosing errors).

[0913] Any adverse and unexpected signs (including abnormal laboratory findings), symptoms, or diseases temporally associated with the use of the drug product or investigational product, whether or not the signs, symptoms, or diseases are related to the drug product or investigational product.

[0914] Clinical laboratory tests

[0915] Additional tests may be performed at any time during the study. Non-limiting examples of such tests are outlined in Table 42.

[0916] Table 42: Clinical laboratory parameters

[0917]

[0918] Abbreviations: HIV = human immunodeficiency virus; SoA = schedule of activities; WOCBP = women of childbearing potential.

[0919] a Trypsinogen will be evaluated only at baseline (for all subjects) and if the subject experiences anaphylaxis within 4 hours after the end of the infusion (samples should be collected as soon as reasonably possible after treatment).

[0920] Adverse events of special interest (AESI)

[0921] For AEs (serious or non-serious) that are scientific and medical concerns of the sponsor-specific investigational drug or program, it may be appropriate for the investigator to continuously monitor them and communicate rapidly with the sponsor. Further research may be required for AESIs to characterize and understand the said AESIs. Depending on the nature of the AESIs, rapid communication between the sponsor and the regulatory agency may also be required.

[0922] Biomarker

[0923] Whole blood samples will be collected to measure exploratory circulating biomarkers according to Table 60. Instructions for the collection and processing of biological samples will be provided by the sponsor in a separate laboratory manual. The actual date and time (24-hour time) of each sample will be recorded in the eCRF. Data collected from the analysis of circulating biomarkers can be used to correlate exposure with safety or efficacy aspects related to concerns that arise during or after the study.

[0924] Circulating biomarkers will be measured using qualified assay methods suitable for the intended use under the supervision of the sponsor or a qualified designee. The analytical methods used to measure these biomarker endpoints will be described in a separate bioanalytical report. Samples will be analyzed only within the window of sample stability.

[0925] Circulating biomarkers to be evaluated include ACE, interleukin-6 (IL-6), sIL 2R, soluble tumor necrosis factor α (sTNFα), C-reactive protein (CRP), calcitriol, and IL-1b. ACE is a common biomarker used in the diagnosis of sarcoidosis. sIL 2R can be elevated in some sarcoidosis subjects as a surrogate marker of T cell activation and shows high and superior sensitivity and specificity for diagnosing sarcoidosis in subjects compared to ACE. IL-6 is typically elevated in subjects with sarcoidosis and can participate in the initiation and maintenance of alveolitis by activating and causing proliferation of T cells. TNFα induces IL-6 through the nuclear factor k B pathway and, thus, inhibition of TNFα by XTMAB-16 may be reflected in changes in the circulating levels of IL-6. Acute tissue injury in the lung induced by IL-1b is transformed into fibrotic changes. IL-1b can be a target for therapeutic intervention in diseases associated with fibrosis and tissue remodeling, such as sarcoidosis.

[0926] Immunogenicity

[0927] Antibodies against XTMAB-16 will be evaluated in serum samples collected from all subjects according to Table 60. Additionally, at the safety follow-up visit, serum samples should also be collected from subjects who discontinue the study or elect not to roll over to the long-term open-label safety study. The persistence of the ADA status will be measured as defined below.

[0928] Transient ADA response:

[0929] (i) Treatment-induced ADA is detected at only one sampling time point during the treatment or follow-up observation period (excluding the last sampling time point, where detection at the last sampling time point should be considered persistent unless shown to be undetectable at a later time), or

[0930] (ii) Treatment-induced ADA is detected at two or more sampling time points during treatment (including the follow-up period, if any), where the first ADA-positive sample and the last ADA-positive sample (regardless of any intervening negative samples) are separated by a period of less than 16 weeks, and the subject's last sampling time point is ADA-negative.

[0931] Persistent ADA response:

[0932] (i) Treatment-induced ADA is detected at two or more sampling time points during treatment (including the follow-up period, if any), where the first ADA-positive sample and the last ADA-positive sample (regardless of any intervening negative samples) are separated by a period of 16 weeks or longer, or

[0933] (ii) Treatment-induced ADA incidence occurs only at the last sampling time point of the treatment study period or at a sampling time point less than 16 weeks before the last sample that is ADA-negative.

[0934] Instructions for the collection and handling of biological samples will be provided by the sponsor in a separate laboratory manual. These samples will be tested by the sponsor or the sponsor's designee. Each serum sample will be divided into 2 aliquots (1 primary and 1 backup each).

[0935] Serum samples will be tested using a multi-tiered approach. Serum samples will initially be evaluated using a validated screening assay for antibodies that bind to XTMAB-16. Samples determined to be presumptively positive for ADA in the screening assay will then be subjected to a confirmatory assay to demonstrate that the antibody is specific for XTMAB-16. Samples identified as positive in the confirmatory assay will be further characterized in an empirical tier assay, and the titer of the confirmed positive samples will be reported. Additional analyses may be performed to verify antibody stability, to study the treatment and / or to further characterize the immunogenicity of the investigational treatment. Serum samples will be screened for antibodies that bind to XTMAB-16, and the titer of the confirmed positive samples will be reported.

[0936] Antibodies against XTMAB-16 will be assayed and characterized using validated assays under the supervision of the Sponsor or a qualified designee. The analytical methods used to measure immunogenicity will be described in a separate biometric report. Samples will be assayed only within the window of sample stability.

[0937] The ability of samples with a positive titer that are confirmed positive for antibodies binding to XTMAB-16 to neutralize the activity of the study treatment can be further characterized and / or evaluated using validated assays.

[0938] Summary of Statistical Considerations

[0939] Unless otherwise specified, summary statistics for continuous variables will consist of the number of non-missing observations (n), mean, standard deviation (SD), minimum, median, and maximum. For categorical variables, the number and percentage of subjects in each category will be presented. Where appropriate, 95% confidence intervals (CIs) will be provided. Unless otherwise stated, the primary analysis set for Part A is the SAS, and the primary analysis set for Part B is the FAS.

[0940] Data will be summarized in tabular form by treatment group for the cross-group combinations in Part A and by treatment group in Part B. In Part A, 2 placebo subjects in each group will be combined into a placebo group consisting of 8 subjects.

[0941] Baseline will be defined as the last non-missing value (including planned and unplanned assessments) before the subject receives study treatment.

[0942] The baseline for corticosteroid (prednisone or equivalent form) dose will be the dose value established at Day 1 at the time of receiving study treatment.

[0943] General Calculation or Derivation of Efficacy Variables

[0944] Percentage reduction of prednisone dose relative to baseline:

[0945] The percentage reduction of prednisone or equivalent form dose at a given time point relative to baseline, which is defined as: {(baseline dose - dose at time point) / baseline dose} * 100%

[0946] Proportion of subjects using prednisone dose ≤ 5 mg / day:

[0947] For each treatment group at a specific time point, the number of subjects using prednisone or equivalent form dose ≤ 5 mg / day will be calculated. The proportion of such subjects in each treatment group will be calculated at the given time point as follows: (number of subjects using prednisone or equivalent form dose ≤ 5 mg / day at the given time point / number of subjects in the treatment group)

[0948] The proportion of subjects with a ≥ 50% reduction in prednisone dose relative to baseline:

[0949] For an individual subject, if the above calculation yields a value of 50% or greater, the subject will be classified as having at least a 50% reduction in the daily prednisone or equivalent dose. The proportion of such subjects in each treatment group will be calculated as follows: (Number of subjects with a ≥ 50% reduction at a given time point / Number of subjects in the treatment group).

[0950] Analysis of the primary endpoint: Part A (MAD)

[0951] There is no primary efficacy endpoint in Part A of the study.

[0952] To observe the primary safety endpoint, the frequency of treatment-emergent adverse events (TEAEs) and the number of subjects with TEAEs will be summarized overall for the treatment groups across the cohorts in Part A in the SAS population and for applicable categories (e.g., severity, treatment-related, seriousness). The observed DLT and AESI rates will also be summarized by dose level and frequency.

[0953] PK, PD, ADA, reduction in corticosteroid dose, QoL, and lung data will be observed and summarized narratively to determine the recommended Phase 2 dose for Part B.

[0954] For the PK endpoint, the plasma concentration of XTMAB-16 will be summarized in tabular form for at least the minimum key PK parameters of interest (C 谷 and AUC) within the dosing interval. The PK methods and analysis will be described in a separate PK analysis plan.

[0955] For the PD endpoint, the biomarkers at each biomarker sampling time point will be summarized narratively as absolute values and percent changes for ACE, sIL 2R, CRP, IL-1b, and calcitriol.

[0956] The reduction in corticosteroid dose will be summarized narratively as absolute values and percent changes over time.

[0957] Quality of life and lung data will be summarized narratively as absolute values and percent changes relative to baseline over time to obtain the mean percent change in FVC%, KSQ general, KSQ-L score, LSQ, and STQ score.

[0958] Analysis of the primary endpoint: Part B (POC)

[0959] For the primary efficacy endpoint of Part B of the study [targeted tapered dose of corticosteroids (maximum dose of prednisone 5 mg / day or equivalent), the 1-sided 95% confidence interval (CI) method for the difference in proportions of subjects with a targeted tapered dose of corticosteroids by Week 12 (using the point estimate of the difference between XTMAB-16 and placebo) will be used to test the superiority of XTMAB-16 in the FAS population. The proportion and its 95% CI obtained using the Clopper–Pearson test will also be shown. A Cochran Mantel Haenszel test (CMH) stratified by the stratifying factor (baseline dose of prednisone or equivalent) will also be conducted. This analysis will be performed in the FAS population.

[0960] The categories of response are defined as follows:

[0961] Responder: For subjects who require corticosteroid rescue therapy between baseline and Week 12, it can be continued to taper off at the discretion of the investigator after the acute event has resolved. Tapering of steroids must be completed by Week 12, and regardless of whether corticosteroid rescue therapy is given, they are considered responders.

[0962] Non-responder: Failure to taper off by Week 12 after rescue therapy will be considered a non-responder.

[0963] Estimand Framework

[0964] The appendix of the International Council for Harmonization (ICH) E9(R1) introduced the concept of the estimand, which transforms the trial objective into a precise definition of the treatment effect to be estimated. For the primary endpoint of Part B, the description of the estimand includes four attributes: the population, the variable (or endpoint) to be obtained for each subject, the description of how to handle concomitant events (ICE), and the population-level summary of the variable. These attributes are described in Table 43.

[0965] Table 43: Estimand Framework

[0966]

[0967] The categories of response in the estimand framework are defined as follows:

[0968] Responder: For subjects who require corticosteroid rescue therapy between baseline and week 12, tapering may continue after the acute event resolves at the discretion of the investigator. Tapering of steroids must be completed by week 12 and the subject is considered a responder regardless of whether corticosteroid rescue therapy was administered.

[0969] Non-responder: In the event that the subject has any of the following reported events, failure to achieve steroid tapering by week 12 will be considered a non-responder:

[0970] i. Receiving more than one course of corticosteroid rescue therapy

[0971] ii. Extrapulmonary disease requiring systemic rather than topical skin and eye treatment

[0972] iii. Increase in background sarcoidosis concomitant medications noted in inclusion criterion 6

[0973] iv. Discontinuation of study drug for any reason

[0974] Analysis of secondary endpoints: Part A (MAD)

[0975] The efficacy, PK, biomarker, and immunogenicity secondary endpoints for Part A will be summarized.

[0976] (i) Efficacy:

[0977] The number and proportion of subjects who achieve targeted tapering of corticosteroids (maximum dose of prednisone 5 mg / day or equivalent) and those who achieve at least a 50% reduction in corticosteroid dose by week 12 will be summarized by treatment group and by pooling the active treatment groups in Part A. The proportion and its 95% CI using the Clopper-Pearson test will also be shown.

[0978] The absolute value of the corticosteroid dose reduction and the change relative to baseline will also be summarized narratively by treatment group and by pooling the active treatment groups in Part A over time.

[0979] All efficacy analyses will be conducted in the FAS population.

[0980] (ii) Pharmacokinetics:

[0981] Plasma concentrations of XTMAB-16 will be analyzed by non-compartmental and / or population PK methods, and the analysis will be described in a separate PK analysis plan. At a minimum, the key PK parameters of interest (C 谷 and AUC over the dosing interval) will be estimated and summarized narratively. Additional evaluations may include assessment of dose proportionality and estimation of time to steady state.

[0982] Stratification by the key covariates of interest (e.g., ADA-confirmed positive status) may be considered when data permit.

[0983] (iii) Biomarkers:

[0984] Each biomarker will be summarized at each biomarker sampling time point by descriptive statistics (N, mean, SD, coefficient of variation (CV)%, geometric mean, geometric CV%, median, minimum, and maximum), and the biomarkers will be tabulated over time by treatment group. Biomarker-time profile plots of individual means and combined means (+ / - SD) will be generated where appropriate.

[0985] Absolute values and percent changes of biomarkers from baseline to EOI, Week 2, Week 4, Week 8, and Week 12 for the following parameters: IL-6, sTNFα.

[0986] (iv) Immunogenicity:

[0987] Anti-drug antibodies and nAbs at each sampling time point will be summarized by treatment group by descriptive statistics (N, mean, SD, CV, geometric mean, geometric CV, median, minimum, and maximum).

[0988] The number and percentage of subjects testing positive for XTMAB-16ADA at baseline, Week 4, Week 8, and Week 12 and the number and percentage testing positive for XTMAB-16nAb at baseline, Week 4, Week 8, and Week 12 will be shown by treatment group. Transient and persistent positive status will also be summarized by number and percentage for follow-up assessment.

[0989] Analysis of secondary endpoints: Part B (POC)

[0990] The safety, efficacy, PK, biomarker, and immunogenicity secondary endpoints of Part B will be summarized.

[0991] (i) Efficacy:

[0992] The difference in proportions of subjects achieving at least a 50% reduction by Week 12 and the difference in proportions of subjects able to maintain steroid reduction at Week 24 will be statistically analyzed similar to the primary endpoint analysis by treatment group in Part B. The proportions and their 95% CIs using the Clopper-Pearson test will also be shown.

[0993] The absolute value of the reduction in corticosteroid dose and the change relative to baseline will also be narratively summarized over time. The number and percentage of targeted stepwise reduced doses of corticosteroids (maximum prednisone 5 mg / day or equivalent) within 12 weeks forward time points will also be summarized by treatment group.

[0994] The analysis will be conducted in the FAS population.

[0995] Prior to the 12-week taper assessment for the primary efficacy endpoint, excluding treatment discontinuation, a sensitivity analysis can be performed.

[0996] (ii) Pharmacokinetics:

[0997] The plasma concentration of XTMAB-16 will be analyzed by non-compartmental and / or population PK methods, and the analysis will be described in an independent PK analysis plan. At a minimum, the key PK parameters of interest (C 谷 and AUC within the dosing interval) will be estimated and narratively summarized. Additional evaluations may include assessment of dose proportionality and the estimated time to reach steady state.

[0998] Stratification by key covariates of interest (e.g., ADA-confirmed positive status) may be considered when the data permit.

[0999] (iii) Biomarkers:

[1000] The biomarkers at each biomarker sampling time point will be summarized by descriptive statistics (N, mean, SD, CV%, geometric mean, geometric CV%, median, minimum, and maximum), and the biomarkers will be tabulated over time. Biomarker-time profile plots of individual means and combined means (±SD) will be generated when appropriate.

[1001] The absolute values and percent changes in biomarkers from baseline to weeks 12 and 24 for the following parameters: ACE, IL-6, sIL 2R, sTNFα, CRP, calcitriol (vitamin D1,25), IL-1b.

[1002] (iv) Immunogenicity:

[1003] The anti-drug antibodies and nAbs at each sampling time point will be summarized by descriptive statistics (N, mean, SD, CV, geometric mean, geometric CV, median, minimum, and maximum).

[1004] The number and percentage of subjects who test positive for XTMAB-16 ADA from baseline to week 24 and the count and percentage of those who test positive for XTMAB-16 nAb from baseline to week 24 will be presented. The transient and persistent positive statuses will also be summarized by count and percentage for follow-up evaluation.

[1005] Analysis of exploratory endpoints: Part A (MAD)

[1006] The quality of life, lung, and imaging exploratory endpoints in Part A will be summarized. The SAS population will be analyzed by treatment group.

[1007] (i) Quality of life (QoL):

[1008] Descriptive summaries of absolute values and changes relative to the baseline QoL endpoints will be summarized by treatment group and over time by combining the active treatments in Part A.

[1009] The list of QoL assessments is as follows. For the order of assessments, see the eCOA site manual: PGA, Fatigue Assessment Scale (FAS), mMRC Dyspnea Scale, SF-36.

[1010] (ii) Lung and imaging:

[1011] Descriptive summaries of lung and imaging endpoints will be summarized over time. Changes relative to the baseline will also be summarized by treatment group and by combining the active treatments in Part A. Assessments are as follows: 6MWD, HRCT.

[1012] Analysis of exploratory endpoints: Part B (POC)

[1013] The quality of life, lung, and imaging exploratory endpoints in Part B will be summarized. The analysis will be by treatment within the FAS population.

[1014] (i) Quality of life (QoL):

[1015] Descriptive summaries of absolute values and changes relative to the baseline QoL endpoints will be summarized over time.

[1016] The list of QoL assessments is as follows. KSQ General, KSQ Lung Module, LCQ, PGA, Fatigue Assessment Scale (FAS), mMRC Dyspnea Scale, SF-36, STQ.

[1017] (ii) Lung and imaging:

[1018] Descriptive summaries of lung and imaging endpoints will be summarized over time. Changes relative to the baseline will also be summarized. Here is the list of assessments: FVC%, KSQ Lung Score, 6MWD, HRCT.

[1019] Physical examinations and vital signs

[1020] Physical examinations will be narratively summarized by visit. Changes relative to the baseline of vital sign observations and parameters will be summarized by subject over time. Clinically notable vital signs (meeting predefined criteria specified in the SAP) of visit subjects with significant vital sign data will be listed.

[1021] Electrocardiogram

[1022] Summarize the changes (units) relative to baseline of electrocardiogram (ECG) observations and parameters over time. Echocardiogram data will be summarized narratively. Echocardiogram data of the overall worst post-baseline values will also be summarized narratively.

[1023] Clinical laboratory evaluations

[1024] Descriptive statistics of clinical laboratory values will be presented for each clinical laboratory assessment. Observations and changes relative to baseline of hematology, blood chemistry, and urine analysis assessments will be summarized over time. In cases where test results are reported in a categorical format, results will be summarized by subject count and percentage for each category.

[1025] Each hematology, blood chemistry, and urine analysis parameter will be classified as low, normal, high, missing based on the reference range. Shift tables related to the normal range of changes relative to baseline over time will be presented. Clinically notable laboratory values will be submitted by visit (if any) based on predefined criteria specified in the SAP.

[1026] A list of laboratory data for individual subjects will be generated. Values meeting predefined criteria that are clinically notable will be flagged as needed within the list.

[1027] Baseline descriptive statistics

[1028] Summarization and analysis of demographics and baseline characteristics, medical history, and prior and concomitant medications will be based on the SAS population in Part A and the ITT in Part B. Medical history data will be listed by subject, including visit, description of disease / procedure, MedDRA, and prior and concomitant medications will be coded according to the latest version of the World Health Organization Drug Dictionary (WHO-DD).

[1029] Subject disposition and protocol deviations are based on all subjects who have signed the ICF in both Part A and Part B.

[1030] Unless otherwise specified, summary statistics for continuous variables will consist of the number (n) of non-missing observations, mean, SD, minimum, median, and maximum. For categorical variables, the number and percentage of subjects in each category will be presented. A list of demographics and baseline characteristics, disposition, protocol deviations, and randomization list (treatment to which the subject was randomly assigned) will also be provided.

[1031] Planned interim analysis

[1032] Interim analysis for Part B: No formal interim analysis is planned for this study. If needed, a provisional review plan will be defined based on the feedback from the DSMC.

[1033] Subgroup analysis: No subgroup analysis is planned for this study.

[1034] Sample size determination: Part A (MAD)

[1035] Thirty-two (32) subjects will be enrolled in Part A to receive XTMAB-16 or placebo in a MAD manner in four cohorts for 12 weeks:

[1036] Cohort 1: 2 mg / kg Q4W for 12 weeks (n = 6 active, 2 placebo)

[1037] Cohort 2: 4 mg / kg Q4W for 12 weeks (n = 6 active, 2 placebo)

[1038] Cohort 3: 2 mg / kg Q2W for 12 weeks (n = 6 active, 2 placebo)

[1039] Cohort 4: 4 mg / kg Q2W for 12 weeks (n = 6 active, 2 placebo)

[1040] Each cohort will enroll 8 subjects. The DSMC will conduct regular reviews of the safety, tolerability, PK, and immunogenicity obtained from the study according to the DSMC charter to provide guidance on whether to increase or decrease the dose and frequency of XTMAB-16.

[1041] Sample size determination: Part B (POC)

[1042] For Part B, a total of 62 subjects will be randomly assigned 1:1 to XTMAB-16: placebo for 24 weeks to demonstrate a statistically significant difference in support of XTMAB-16. Assuming that the proportion of subjects in the placebo group who reach the targeted tapered dose of corticosteroids (maximum dose of prednisone 5 mg / day or equivalent) by week 12 is approximately 15% and the expected proportion in the XTMAB-16 group is 45%, i.e., the treatment difference is approximately 30%, a total of 56 subjects are needed to demonstrate this benefit at week 12 with a one-sided α = 0.05 and 80% power using the CMH test with stratification factors to test the proportion (baseline dose of prednisone or equivalent). Considering an approximately 10% dropout, a total of 62 randomly assigned subjects are sufficient for the POC part of this study.

[1043] Schedule of Activities (SoA)

[1044] Part A (MAD) of the study is an independent 12-week sequential multiple ascending dose study. Due to the limited sample size of each treatment group, no formal statistical hypotheses will be tested in Part A of this study.

[1045] Part B (POC) is an independent double-blind placebo-controlled proof-of-concept study. To confirm the preliminary efficacy of XTMAB-16 at week 12, the following hypotheses will be tested in Part B of this study:

[1046] H0: p x - p p = 0, H1: p x - p p > 0

[1047] p x and p p represent, respectively, the proportions of subjects in the XTMAB-16 group and the placebo group who have achieved a targeted progressive reduction in corticosteroid dose (prednisone ≤ 5 mg / day dose or equivalent) by week 12.

[1048] Table 44: Activity schedule, 1st of 4 – Part A (MAD)

[1049]

[1050]

[1051] Table 45: Activity schedule, 2nd of 4 – Part A (MAD)

[1052]

[1053] Table 46: Activity schedule, 3rd of 4 – Part A (MAD)

[1054]

[1055] Table 47: Activity schedule, 4th of 4 – Part A (MAD)

[1056]

[1057] Abbreviations: 6MWD = 6-minute walk distance; ACE = angiotensin-converting enzyme; ADA = anti-drug antibody; AE = adverse event; BL = baseline; COVID-19 = coronavirus disease; CRP = C-reactive protein; CT = computed tomography; D = day; ECG = electrocardiogram; EOT = end of treatment; FSH = follicle-stimulating hormone; FVC = forced vital capacity; HBsAg = hepatitis B surface antigen; HCV = hepatitis C virus; HIV = human immunodeficiency virus; HRCT = high-resolution computed tomography; IgG = immunoglobulin G; IL-1b = interleukin-1b; IL-6 = interleukin-6; IRR = infusion-related reaction; MAD = multiple ascending dose; mMRC = modified Medical Research Council; nAb = neutralizing antibody; PCR = polymerase chain reaction; PD = pharmacodynamics; Q2W = every 2 weeks; Q4W = every four weeks; SAE = serious adverse event; SCR = screening; sIL 2R = soluble interleukin-2 receptor; sTNFα = soluble tumor necrosis factor α; TB = tuberculosis; Unsch = unscheduled; W = week.

[1058] a The screening period is up to 42 days from the time of informed consent to enrollment.

[1059] b In the case that the subject completes the EOT assessment at Week 12, the Day 0 assessment at Week 12 does not need to be repeated.

[1060] c To evaluate infusion site reactions, the infusion site should be examined at 0.5 hour and 1.5 hours (±10 minutes) after the end of infusion. The subject's IRR and allergies should be monitored during this time period.

[1061] d A complete physical examination will be performed during screening, EOT, and all dosing days. A brief physical examination will be performed at the visit without dosing.

[1062] Targeted physical examinations can be performed at any time at the discretion of the investigator. Height is measured at screening; and weight is measured at each visit.

[1063] e A serum pregnancy test will be performed at screening and a urine pregnancy test will be performed thereafter.

[1064] f The tryptase level of all subjects will be evaluated at baseline. For any subject who experiences an allergy within 4 hours after the end of infusion, additional samples should be collected as soon as reasonably possible to evaluate the tryptase level.

[1065] g A standard 12-lead ECG will be recorded. The ECG will be collected in triplicate during screening and at baseline, with approximately 1 minute between each ECG; and all other ECGs will be collected as a single record.

[1066] h COVID-19 testing will be completed by a rapid antigen test or PCR test performed locally.

[1067] i Containing only known potentially significant fibrotic disease and / or active inflammation in the hilar region, as shown by HRCT and confirmed by a central reader. Subjects with current active inflammation in the hilar region and concurrent inflammation outside the hilar region may be included. For subjects with disease onset < 2 years, historical CT confirmed by central reading performed within 6 months before screening is acceptable. For subjects with disease onset > 2 years and no CT performed within 6 months before screening, CT will be performed at screening.

[1068] j HRCT scans will be performed at baseline ± 10 days.

[1069] k All quality of life assessments should be performed before other study procedures at each scheduled visit and should be completed in the same order at each scheduled visit.

[1070] l See Table 60 for pharmacokinetics, anti-drug antibody, and biomarker collection time points.

[1071] Table 48: Activity Schedule, 1 of 4 – Part B (POC)

[1072]

[1073]

[1074] Table 49: Activity Schedule, 2 of 4 – Part B (POC)

[1075]

[1076]

[1077] Table 50: Activity Schedule, 3 of 4 – Part B (POC)

[1078]

[1079] Table 51: Activity Schedule, 4 of 4 – Part B (POC)

[1080]

[1081] Abbreviations: 6MWD = 6-minute walk distance; ACE = angiotensin-converting enzyme; ADA = anti-drug antibody; AE = adverse event; COVID-19 = coronavirus disease;

[1082] CRP = C-reactive protein; D = day; ECG = electrocardiogram; EOT = end of treatment; FSH = follicle-stimulating hormone; FVC = forced vital capacity; HBsAg = hepatitis B surface antigen; HCV = hepatitis C virus; HIV = human immunodeficiency virus; HRCT = high-resolution computed tomography; IgG = immunoglobulin G; IL-1b = interleukin 1b; IL-6 = interleukin-6; IRR = infusion-related reaction; mMRC = modified Medical Research Council; nAb = neutralizing antibody; PCR = polymerase chain reaction; PD = pharmacodynamics; Q2W = every two weeks; Q4W = every four weeks; SAE = serious adverse event; SCR = screening; sIL 2R = soluble interleukin-2 receptor; sTNFα = soluble tumor necrosis factor α; TB = tuberculosis; Unsch = unscheduled; W = week.

[1083] a The screening period is up to 42 days from the informed consent to enrollment.

[1084] b In the case where the subject completes the EOT assessment at week 24, the week 24 day 0 assessment does not need to be repeated.

[1085] c To evaluate infusion site reactions, the infusion site should be examined at 0.5 hour and 1.5 hours (±10 minutes) after the end of the infusion. The subject's IRR and allergies should be monitored during this time period.

[1086] d A complete physical examination will be performed during screening, EOT, and all dosing days. A brief physical examination will be performed at the visit without dosing.

[1087] Targeted physical examinations can be performed at any time at the discretion of the investigator. Height is measured at screening; and weight is measured at each visit.

[1088] e A serum pregnancy test will be performed at screening and a urine pregnancy test will be performed thereafter.

[1089] f The tryptase levels of all subjects will be evaluated at baseline. For any subject who experiences an allergy within 4 hours after the end of the infusion, additional samples should be collected as soon as reasonably possible to evaluate the tryptase levels.

[1090] g A standard 12-lead ECG will be recorded. The ECG will be collected as a single recording.

[1091] h COVID-19 testing will be completed by a locally performed rapid antigen test or PCR test.

[1092] i An HRCT scan will be performed at baseline ±10 days.

[1093] All quality of life assessments should be conducted prior to other study procedures at each scheduled visit and should be completed in the same order at each scheduled visit.

[1094] kSee Table 60 for pharmacokinetic, anti-drug antibody, and biomarker collection time points.

[1095] Table 52: Pharmacokinetic, anti-drug antibody, and biomarker collection time points

[1096]

[1097]

[1098] Abbreviations: ADA = anti-drug antibody; EOI = end of infusion; EOT = end of treatment; nAb = neutralizing antibody; PK = pharmacokinetics; Q2W = every two weeks; Q4W = every four weeks; SOI = start of infusion.

[1099] aThe sampling window is -30 minutes.

[1100] bThe sampling window is ±5 minutes.

[1101] cThe sampling window is ±3 hours.

[1102] dRespectively, the 12-week and 24-week visits in Parts A and B are EOT visits and are not safety follow-up visits for non-rolling subjects.

[1103] Scientific rationale for the study design

[1104] Part A of this study will evaluate the safety, pharmacokinetics, pharmacodynamics, effects on inflammatory biomarkers, and potential efficacy of four different multiple-dose regimens (mg / kg dose and administration frequency) that vary in the predicted mean and trough concentrations of XTMAB-16, enabling identification of the optimal regimen for subjects with sarcoidosis with or without pulmonary manifestations.

[1105] Within Part B, the study will confirm the preliminary efficacy of XTMAB-16 (as measured by the ability to reduce the use of background oral corticosteroids in subjects with pulmonary sarcoidosis with or without extrapulmonary manifestations) and evaluate multiple clinically relevant endpoints.

[1106] Dose rationale is based in part on the results of Example 8.

[1107] Table 53: Conditions for repeated administration of the stimulus

[1108]

[1109] Abbreviations: ADA = anti-drug antibody; AUC tau = area under the curve; C 平均值 = mean concentration; Ceoi = concentration at the end of infusion; C 最大值 = maximum concentration; C 谷 = trough concentration; kg = kilometer; mg = milligram; MRT = mean residence time; Q2W = every 2 weeks; Q4W = every 4 weeks; t 1 / 2 = half-life; T 最大值 = time to reach the maximum observed concentration; V ss = apparent volume of distribution at steady state.

[1110] Ceoi is set relative to the simulated serum XTMAB-16 concentration at the nominal time of 2 hours. Tau = 336 hours after Q2W dosing, Tau = 672 hours after Q4W dosing

[1111] Definition of study end

[1112] In the case where a subject has completed all study periods including the last visit or the last planned procedure shown in the Schedule of Activities (SoA), the subject will be considered to have completed the study (Tables 52 - 59).

[1113] All subjects who complete Part A or Part B will participate in a long-term open-label follow-up study at the dose and frequency established in Part A.

[1114] The end of treatment (EOT) for Part A will be considered the last dose at week 12. Subjects who do not choose to participate in the open-label extension will complete safety follow-up visits through week 20 as outlined in the SoA (Tables 52 - 59).

[1115] The EOT for Part B will be considered the last dose at week 24. Subjects who do not choose to participate in the open-label extension will complete safety follow-up visits through week 34 as outlined in the SoA (Tables 44 - 51).

[1116] The end of the study will be defined as the date of the last visit of the last subject in the study or the last planned procedure of the last subject in the overall trial as shown in the SoA.

[1117] The teachings of the inventive techniques provided herein can be applied to other methods, not necessarily the methods described above. The elements and acts of the various embodiments described above can be combined to provide additional embodiments.

[1118] All of the above patents and applications and other references (including any literature that may be listed in the accompanying file) are hereby incorporated by reference. Where necessary, various aspects of the technology of the present invention may be modified to adopt the methods, functions, and concepts of the various references described above to provide further additional embodiments of the technology of the present invention.

[1119] In view of the description detailed above, these and other changes may be made to the technology of the present invention. Although the above description details certain embodiments of the technology of the present invention and describes the best mode contemplated, the technology of the present invention may be practiced in many ways, no matter how detailed the above text may appear. The details of the methods may vary quite significantly in their specific implementation details while still being covered by the technology of the present invention.

[1120] As noted above, the specific terms used in describing certain features or aspects of the technology of the present invention should not be construed as implying that such terms are re - defined herein to be limited to any specific characteristics, features, or aspects of the technology of the present invention associated with such terms. Generally, the terms used in the following claims should not be construed to limit the technology of the present invention to the specific embodiments of the technology of the present invention unless such terms are explicitly defined in the detailed description above. Thus, the actual scope of the technology of the present invention not only covers the embodiments of the technology of the present invention but also covers all equivalent ways of practicing or implementing the technology of the present invention under the claims.

[1121] In view of the foregoing, it should be understood that specific embodiments of the technology of the present invention have been described for purposes of illustration, but various changes may be made without departing from the scope of the technology of the present invention. Thus, the technology of the present invention is not limited except as restricted by the appended claims.

[1122] Additional embodiments

[1123] The technology of the present invention includes, but is not limited to, the following specific embodiments set forth in paragraphs

[0753] -

[1031] below:

[1124] 1. A method for treating or reducing sarcoidosis in a subject in need thereof relative to a control treatment, the method comprising administering to the subject a first dose of a composition at about 0.5 mg / kg to about 6.0 mg / kg about once every four weeks for a first period of time, the composition comprising an anti - TNFα antibody sialylated with one or more N - acetylneuraminic acid molecules.

[1125] 2. A method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every two weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[1126] 3. A method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every four weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[1127] 4. A method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every two weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

[1128] 5. The method according to any one of embodiments 1 to 4, wherein the first period of time is about 6 weeks to about 24 weeks.

[1129] 6. The method according to embodiment 5, wherein the first period of time is about 8 weeks.

[1130] 7. The method according to embodiment 5, wherein the first period of time is about 12 weeks.

[1131] 8. The method according to embodiment 5, wherein the first period of time is about 24 weeks.

[1132] 9. The method according to any one of embodiments 1 to 8, wherein the first dose is about 2 mg / kg of the composition.

[1133] 10. The method according to any one of embodiments 1 to 8, wherein the first dose is about 4 mg / kg of the composition.

[1134] 11. The method according to any one of embodiments 1 to 10, wherein the anti-TNFα antibody comprises complementarity determining regions (CDRs) that are 100% identical to SEQ ID NO: 3, 4, 5, 6, 7, 8, 10, 12, 13, 14, or 15 or YA.

[1135] 12. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control treatment, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules;

[1136] The first dose comprises about 2 mg / kg of the composition administered once every four weeks for a first period of time, and

[1137] The anti-TNFα antibody comprises:

[1138] (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 3;

[1139] (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 4;

[1140] (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 5;

[1141] (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 6;

[1142] (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 7;

[1143] (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 8;

[1144] (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 10;

[1145] (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: YA;

[1146] (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 12;

[1147] (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 13;

[1148] (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 14; and

[1149] (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 15.

[1150] 13. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules;

[1151] The first dose comprises about 2 mg / kg of the composition administered once every two weeks during a first time period; and

[1152] The anti-TNFα antibody comprises:

[1153] (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 3;

[1154] (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 4;

[1155] (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 5;

[1156] (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 6;

[1157] (v) A fifth CDR, wherein the fifth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:7;

[1158] (vi) A sixth CDR, wherein the sixth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:8;

[1159] (vii) A seventh CDR, wherein the seventh CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:10;

[1160] (viii) An eighth CDR, wherein the eighth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to YA;

[1161] (ix) A ninth CDR, wherein the ninth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:12;

[1162] (x) A tenth CDR, wherein the tenth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:13;

[1163] (xi) An eleventh CDR, wherein the eleventh CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:14; and

[1164] (xii) A twelfth CDR, wherein the twelfth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:15.

[1165] 14. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules;

[1166] The first dose comprises about 4 mg / kg of the composition administered once every four weeks for a first period of time; and

[1167] The anti-TNFα antibody comprises:

[1168] (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3;

[1169] (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4;

[1170] (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5;

[1171] (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6;

[1172] (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7;

[1173] (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8;

[1174] (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10;

[1175] (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA;

[1176] (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12;

[1177] (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13;

[1178] (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and

[1179] (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:15.

[1180] 15. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules;

[1181] The first dose comprises about 4 mg / kg of the composition administered once every two weeks for a first period of time; and

[1182] The anti-TNFα antibody comprises:

[1183] (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:3;

[1184] (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:4;

[1185] (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:5;

[1186] (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:6;

[1187] (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:7;

[1188] (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:8;

[1189] (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:10;

[1190] (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA;

[1191] (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12;

[1192] (x) A tenth CDR having an amino acid sequence that is at least about 8...

Claims

1. A method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every four weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

2. A method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every two weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

3. A method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every four weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

4. A method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose of a composition at about once every two weeks in an amount of about 0.5 mg / kg to about 6.0 mg / kg for a first period of time, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules.

5. The method according to any one of claims 1 to 4, wherein the first period of time is about 6 weeks to about 24 weeks.

6. The method according to claim 5, wherein the first period of time is about 8 weeks.

7. The method according to claim 5, wherein the first period of time is about 12 weeks.

8. The method according to claim 5, wherein the first period of time is about 24 weeks.

9. The method according to any one of claims 1 to 8, wherein the first dose is about 2 mg / kg of the composition.

10. The method according to any one of claims 1 to 8, wherein the first dose is about 4 mg / kg of the composition.

11. The method according to any one of claims 1 to 10, wherein the anti-TNFα antibody comprises complementarity determining regions (CDRs) that are 100% identical to SEQ ID NO: 3, 4, 5, 6, 7, 8, 10, 12, 13, 14, or 15 or YA.

12. A method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered at about once every four weeks during a first period of time, and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

13. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every two weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

14. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 4 mg / kg of the composition administered once every four weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

15. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 4 mg / kg of the composition administered once every two weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

16. A method of reducing granuloma size or decreasing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every four weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

17. A method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every two weeks during a first time period; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

18. A method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 4 mg / kg of the composition administered once every four weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

19. A method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 4 mg / kg of the composition administered once every two weeks for a first period of time; and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

20. The method according to any one of claims 1 to 19, further comprising administering a second dose of the composition to the subject for a second period of time.

21. The method according to claim 20, wherein the second period of time is from about 6 weeks to about 16 weeks.

22. The method according to claim 21, wherein the second period of time is about 8 weeks.

23. The method according to claim 21, wherein the second period of time is about 12 weeks.

24. The method according to any one of claims 20 to 23, wherein the second dose comprises from about 1 mg / kg to about 5 mg / kg.

25. The method according to claim 24, wherein the second dose is about 2 mg / kg.

26. The method according to claim 24, wherein the second dose is about 4 mg / kg.

27. The method according to claim 24, wherein the second dose is about twice the first dose.

28. The method according to claim 27, wherein the administration frequency of the second dose comprises the same administration frequency as that of the first dose.

29. The method according to claim 27, wherein the administration frequency of the second dose consists of the same administration frequency as that of the first dose.

30. The method according to claim 28 or 29, wherein the first dose and the second dose are each administered about once every four weeks.

31. The method according to claim 20, wherein the second dose is a dose substantially the same as the first dose.

32. The method according to claim 20, wherein the second dose is the same dose as the first dose.

33. The method according to claim 31 or 32, wherein the first dose and the second dose are about 2 mg / kg.

34. The method according to claim 31 or 32, wherein the administration frequency of the second dose is about twice the administration frequency of the first dose.

35. The method according to claim 33 or 34, wherein the first dose is administered about once every four weeks, and the second dose is administered about once every two weeks.

36. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprises about 2 mg / kg of the composition administered once every four weeks during a first time period; and the second dose comprises (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period.

37. A method of reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprises about 2 mg / kg of the composition administered once every four weeks during a first time period; and the second dose comprises (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period.

38. The method according to claim 36 or 37, wherein the first time period is from about 6 weeks to about 12 weeks.

39. The method according to claim 38, wherein the first time period is about 8 weeks.

40. The method according to claim 38, wherein the first time period is about 12 weeks.

41. The method according to any one of claims 36 to 40, wherein the second time period is from about 6 weeks to about 12 weeks.

42. The method according to claim 41, wherein the second time period is about 8 weeks.

43. The method according to claim 41, wherein the second time period is about 12 weeks.

44. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control treatment, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered once every four weeks for a first time period; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks, for a second time period; the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR, the tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 13; (xi) An eleventh CDR, the eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 14; and (xii) A twelfth CDR, the twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:

15.

45. A method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered biweekly for a first period of time; and the second dose comprising (i) about 2 mg / kg of the composition administered biweekly for a second period of time, or (iii) about 4 mg / kg of the composition administered once every four weeks; the anti-TNFα antibody comprising: (i) A first CDR, the first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 3; (ii) A second CDR, the second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 4; (iii) A third CDR, the third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 5; (iv) A fourth CDR, the fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 6; (v) A fifth CDR, the fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 7; (vi) A sixth CDR, the sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 8; (vii) A seventh CDR, the seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 10; (viii) An eighth CDR, the eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

46. A method of reducing granuloma size or decreasing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition, the composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; The first dose comprises about 2 mg / kg of the composition administered once every four weeks during a first time period; The second dose comprises (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every four weeks, during a second time period; The anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

47. A method of reducing granuloma size or decreasing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of a composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules; the first dose comprising about 2 mg / kg of the composition administered biweekly for a first period of time; the second dose comprising (i) about 2 mg / kg of the composition administered biweekly or (iii) about 4 mg / kg of the composition administered every four weeks for a second period of time; the anti-TNFα antibody comprising: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:

15.

48. The method according to any one of claims 20 to 47, further comprising administering (c) a third dose of said composition at about 4 mg / kg every about two weeks for a third period of time.

49. The method according to claim 48, wherein said third period of time is at least about 24 weeks.

50. The method of any one of claims 1 to 49, wherein the anti-TNFα antibody comprises a variable weight (V H ) domain, the V H The domain has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

16.

51. The method according to any one of claims 1 to 49, wherein the anti-TNFα antibody comprises a variable light (V L ) domain, and the V L domain has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

9.

52. The method according to any one of claims 1 to 51, wherein said anti-TNFα antibody comprises an asparagine-linked glycosylation site located on at least one heavy chain in the constant Fc region.

53. The method according to claim 52, wherein said anti-TNFα antibody comprises asparagine-linked glycosylation sites located on both heavy chains in the constant Fc region.

54. The method according to any one of claims 50 to 53, wherein said anti-TNFα antibody is sialylated at Asn 297.

55. The method according to any one of claims 1 to 54, wherein said anti-TNFα antibody does not contain N-glycolylneuraminic acid (Neu5Gc).

56. The method according to any one of claims 1 to 55, wherein said anti-TNFα antibody is an immunoglobulin G (IgG) antibody.

57. The method according to claim 43, wherein said IgG antibody is an IgG1 antibody.

58. The method according to claim 57, wherein said IgG1 antibody comprises a κ isotype.

59. The method according to any one of claims 1 to 58, wherein said anti-TNFα antibody is a chimeric antibody.

60. The method according to any one of claims 1 to 59, wherein said anti-TNFα antibody comprises a C-terminal lysine on at least one heavy chain.

61. The method according to claim 60, wherein said anti-TNFα antibody comprises C-terminal lysines on both heavy chains.

62. The method according to claim 60 or 61, wherein the C-terminal lysine is not trimmed.

63. The method according to any one of claims 1 to 62, wherein the composition is administered by infusion.

64. The method according to claim 63, wherein the infusion is an intravenous (IV) infusion.

65. The method according to claim 63 or 64, wherein the infusion is administered over a period of at least about 2 hours.

66. The method according to claim 63 or 64, wherein the infusion is administered over a period of about 2 hours.

67. The method according to any one of claims 1 to 66, wherein the subject receives a cumulative dose of about 6 mg / kg of the composition during the first period.

68. The method according to any one of claims 1 to 66, wherein the subject receives a cumulative dose of about 8 mg / kg of the composition during the first period.

69. The method according to any one of claims 1 to 66, wherein the subject receives a cumulative dose of about 16 mg / kg of the composition during the first period.

70. The method according to any one of claims 41 to 69, wherein the subject receives a cumulative dose of about 10 mg / kg to about 16 mg / kg of the composition during the second period.

71. The method according to claim 70, wherein the subject receives a cumulative dose of about 10 mg / kg of the composition during the second period.

72. The method according to claim 70, wherein the subject receives a cumulative dose of about 12 mg / kg of the composition during the second period.

73. The method according to claim 70, wherein the subject receives a cumulative dose of about 14 mg / kg of the composition during the second period.

74. The method according to claim 70, wherein the subject receives a cumulative dose of about 16 mg / kg of the composition during the second period.

75. The method according to any one of claims 48 to 74, wherein the subject receives a cumulative dose of about 28 mg / kg of the composition.

76. The method according to any one of claims 1 to 75, wherein the subject is administered a cumulative dose of about 30 mg / kg to about 60 mg / kg of the composition.

77. The method according to claim 76, wherein the subject is administered a cumulative dose of about 46 mg / kg of the composition.

78. The method according to claim 76, wherein the subject is administered a cumulative dose of about 48 mg / kg of the composition.

79. The method according to claim 76, wherein the subject is administered a cumulative dose of about 50 mg / kg of the composition.

80. The method according to claim 76, wherein the subject is administered a cumulative dose of about 52 mg / kg of the composition.

81. The method according to any one of claims 1 to 80, wherein the subject is administered at least 3 doses of the composition.

82. The method according to any one of claims 1 to 81, wherein the subject is administered at least 4 doses of the composition.

83. The method according to any one of claims 1 to 82, wherein the subject is administered at least 6 doses of the composition.

84. The method according to any one of claims 1 to 83, wherein the subject is administered at least 7 doses of the composition.

85. The method according to claim 81, wherein the at least 3 doses are administered over a 4-week period.

86. The method according to claim 81, wherein the at least 3 doses are administered over an 8-week period.

87. The method according to claim 81, wherein the at least 3 doses are administered over a 12-week period.

88. The method according to claim 82, wherein the at least 4 doses are administered over a 9-week period.

89. The method according to claim 82, wherein the at least 4 doses are administered over a 10-week period.

90. The method according to claim 82, wherein the at least 4 doses are administered over an 11-week period.

91. The method according to claim 82, wherein the at least 4 doses are administered over a 12-week period.

92. The method according to claim 83, wherein the at least 6 doses are administered over a 10-week period.

93. The method according to claim 83, wherein the at least 6 doses are administered over a 13-week period.

94. The method according to claim 83, wherein the at least 6 doses are administered over a 16-week period.

95. The method according to claim 83, wherein the at least 6 doses are administered over a 17-week period.

96. The method according to claim 83, wherein the at least 6 doses are administered over a 20-week period.

97. The method according to claim 84, wherein the at least 7 doses are administered over a 12-week period.

98. The method according to claim 84, wherein the at least 7 doses are administered over a 15-week period.

99. The method according to claim 84, wherein the at least 7 doses are administered over a 21-week period.

100. The method according to claim 82, wherein the at least 7 doses are administered over a 28-week period.

101. The method according to claim 84, wherein the at least 7 doses are administered over a 24-week period.

102. The method according to any one of claims 1 to 101, wherein the administration of the composition reduces the level of one or more transcripts selected from the group consisting of angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL6), interleukin 2 receptor (IL2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL1B), interleukin 12 (IL12), interleukin 18 (IL18), interferon γ (IFNG), interleukin 8 (IL8), monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), chemokine ligand 2 (CXCL2), chemokine ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JunB), mitogen-activated protein kinase (MKP1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL32), intercellular adhesion molecule 1 (ICAM-1), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), and vascular cell adhesion molecule 1 (VCAM-1) as compared to a control.

103. The method according to claim 102, wherein the level of the one or more transcripts is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% as compared to the control.

104. The method according to any one of claims 1 to 103, wherein the administration of the composition reduces the level of one or more proteins selected from the group consisting of angiotensin-converting enzyme 2 (ACE2), interleukin 6 (IL-6), interleukin 2 receptor (IL-2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL-1B), interferon γ (IFN-γ), interleukin 8 (IL-8), interleukin 12 (IL-12), interleukin 18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), chemokine ligand 2 (CXCL2), chemokine ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JUNB), mitogen-activated protein kinase phosphatase 1 (MKP-1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL-32), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1), relative to a control.

105. The method according to claim 104, wherein the level of the one or more proteins is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

106. The method according to any one of claims 1 to 105, wherein the administration of the composition increases one or more of the following: forced vital capacity (FVC) measurement results, forced expiratory volume in 1 second (FEV1) measurement results, Leicester Cough Questionnaire (LCQ) score, Short Form 36 Health Survey (SF-36) score, King's Sarcoidosis Questionnaire (KSQ) score, steroid toxicity scale (STS) score, or 6-minute walk test (6MWT) distance, relative to a control.

107. The method according to claim 106, wherein the FVC measurement results are increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

108. The method according to claim 106, wherein the FEV1 measurement result is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

109. The method according to claim 106, wherein the LCQ score is increased by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points or 18 points relative to the control.

110. The method according to claim 106, wherein the SF-36 score is increased by at least 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points or 100 points relative to the control.

111. The method according to claim 106, wherein the KSQ score is increased by at least 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points or 100 points relative to the control.

112. The method according to claim 106 or 111, wherein the KSQ score is the KSQ general health status module or the KSQ lung score.

113. The method according to claim 106, wherein the STS score is increased by at least 1 point, 2 points, 3 points, 4 points, 5 points or 6 points relative to the control.

114. The method according to claim 106, wherein the 6MWT distance is increased by at least 10 m, 20 m, 30 m, 40 m, 50 m, 60 m, 70 m, 80 m, 90 m, 100 m, 125 m, 150 m, 175 m, 200 m, 250 m, 300 m, 350 m, 400 m, 450 m or 500 m relative to the control.

115. The method according to any one of claims 1 to 114, wherein the administration of the composition reduces one or more of the following relative to the control: the patient global assessment (PGA) score, the fatigue assessment scale (FAS) score, the modified medical research council (mMRC) dyspnea scale score, the Saint George's Respiratory Questionnaire (SGRQ) score or the Borg's CR10 dyspnea score.

116. The method according to claim 115, wherein the PGA score is reduced by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points or 10 points relative to the control.

117. The method according to claim 115, wherein the FAS score is reduced by at least about 1 point, 2 points, 3 points, or 4 points relative to the control.

118. The method according to claim 115, wherein the mMRC scale score is reduced by at least about 1 level, 2 levels, 3 levels, or 4 levels relative to the control.

119. The method according to claim 115, wherein the SGRQ score is reduced by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points, or 100 points relative to the control.

120. The method according to claim 115, wherein the Borg CR10 dyspnea score is reduced by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, or 10 points relative to the control.

121. The method according to any one of claims 1 to 120, wherein the administration of the composition reduces the need for anti-inflammatory drugs relative to the control.

122. The method according to claim 121, wherein the anti-inflammatory drug is selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, efzofitimod, rituximab, adalimumab, golimumab, namilumab, and infliximab.

123. The method according to claim 122, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone, and triamcinolone.

124. The method according to any one of claims 121 to 123, wherein the reduced need for the anti-inflammatory drug comprises a reduction in the frequency, dose, or duration of administration of the anti-inflammatory drug.

125. The method according to any one of claims 1 to 123, wherein the administration of the composition reduces granuloma formation or decreases the granuloma size in the subject by about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control.

126. The method according to claim 125, wherein the granuloma is a cutaneous granuloma, pulmonary granuloma, lymph node granuloma, ocular granuloma, hepatic granuloma, splenic granuloma, cardiac granuloma, or rheumatoid granuloma.

127. The method according to any one of claims 1 to 126, wherein the administration of the composition reduces the level of calcitriol in the subject relative to a control.

128. The method according to claim 127, wherein the level of calcitriol is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

129. The method according to any one of claims 1 to 128, wherein the administration of the composition reduces the level of neopterin in the subject relative to a control.

130. The method according to claim 129, wherein the level of neopterin is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

131. The method according to any one of claims 1 to 130, wherein the administration of the composition reduces the level of lysozyme in the subject relative to a control.

132. The method according to claim 131, wherein the level of lysozyme is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

133. The method according to any one of claims 1 to 132, wherein the administration of the composition reduces the soluble IL-2R (sIL-2R) level in the subject relative to a control.

134. The method according to claim 132, wherein the sIL-2R level is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

135. The method according to any one of claims 1 to 134, wherein the subject has one or more of the following: acute sarcoidosis, chronic sarcoidosis, progressive sarcoidosis, pulmonary sarcoidosis, remitting sarcoidosis, refractory sarcoidosis, advanced sarcoidosis, ocular sarcoidosis, cutaneous sarcoidosis or neurosarcoidosis.

136. The method according to any one of claims 1 to 135, wherein the subject has received an anti-inflammatory drug.

137. The method according to any one of claims 1 to 136, wherein the subject is receiving an anti-inflammatory drug.

138. The method according to claim 136 or 137, wherein the anti-inflammatory drug is selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, etrolizumab, rituximab, adalimumab, golimumab, namilumab and infliximab.

139. The method according to claim 138, wherein the corticosteroid is selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone and triamcinolone.

140. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every about four weeks for a first period of time.

141. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every about two weeks for a first period of time.

142. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg once every about four weeks for a first period of time.

143. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 0.5 mg / kg to about 6.0 mg / kg approximately once every two weeks for a first period of time.

144. The composition according to any one of claims 140 to 143, wherein the first period of time is about 6 weeks to about 24 weeks.

145. The composition according to claim 144, wherein the first period of time is about 8 weeks.

146. The composition according to claim 144, wherein the first period of time is about 12 weeks.

147. The composition according to claim 144, wherein the first period of time is about 24 weeks.

148. The composition according to any one of claims 140 to 147, wherein the first dose is about 2 mg / kg of the composition.

149. The composition according to any one of claims 140 to 147, wherein the first dose is about 4 mg / kg of the composition.

150. The composition according to any one of claims 140 to 149, wherein the anti-TNFα antibody comprises complementarity-determining regions (CDRs) that are 100% identical to SEQ ID NO: 3, 4, 5, 6, 7, 8, 10, 12, 13, 14, or 15 or YA.

151. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, wherein the composition is administered at a first dose of about 2.0 mg / kg approximately once every four weeks for a first period of time, and the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

152. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, wherein the composition is administered at a first dose of about 2.0 mg / kg approximately once every two weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

153. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, wherein the composition is administered at a first dose of about 4.0 mg / kg once every about four weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

154. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, wherein the composition is administered at a first dose of about 4.0 mg / kg approximately once every two weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

155. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 2.0 mg / kg once every about four weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

156. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 2.0 mg / kg approximately once every two weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

157. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 4.0 mg / kg approximately once every four weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

158. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, wherein the composition is administered at a first dose of about 4.0 mg / kg approximately once every two weeks for a first period of time, and the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

159. The composition according to any one of claims 140 to 158, wherein the method further comprises administering a second dose of the composition to the subject for a second period of time.

160. The composition according to claim 159, wherein the second period of time is from about 6 weeks to about 16 weeks.

161. The composition according to claim 60, wherein the second period of time is about 8 weeks.

162. The composition according to claim 60, wherein the second period of time is about 12 weeks.

163. The composition according to any one of claims 159 to 162, wherein the second dose comprises from about 1 mg / kg to about 5 mg / kg.

164. The composition according to claim 163, wherein the second dose is about 2 mg / kg.

165. The composition according to claim 163, wherein the second dose is about 4 mg / kg.

166. The composition according to claim 163, wherein the second dose is about twice the first dose.

167. The composition according to claim 166, wherein the frequency of administration of the second dose comprises the same frequency of administration as the first dose.

168. The composition according to claim 166, wherein the frequency of administration of the second dose consists of the same frequency of administration as the first dose.

169. The composition according to claim 167 or 168, wherein the first dose and the second dose are each administered about once every four weeks.

170. The composition according to claim 159, wherein the second dose is a dose substantially the same as the first dose.

171. The composition according to claim 159, wherein the second dose is the same dose as the first dose.

172. The composition according to claim 170 or 171, wherein the first dose and the second dose are about 2 mg / kg.

173. The composition according to claim 170 or 171, wherein the administration frequency of the second dose is about twice the administration frequency of the first dose.

174. The composition according to claim 172 or 173, wherein the first dose is administered about once every four weeks, and the second dose is administered about once every two weeks.

175. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method for treating or reducing sarcoidosis in a subject in need thereof relative to a control, the method comprising administering a first dose and a second dose of the composition to the subject; the first dose comprising about 2 mg / kg of the composition administered once every four weeks during a first time period; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period.

176. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method for reducing the granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering a first dose and a second dose of the composition to the subject; the first dose comprising about 2 mg / kg of the composition administered once every four weeks during a first time period; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period.

177. The composition according to claim 175 or 176, wherein the first time period is about 6 weeks to about 12 weeks.

178. The composition according to claim 177, wherein the first time period is about 8 weeks.

179. The composition according to claim 177, wherein the first time period is about 12 weeks.

180. The composition according to any one of claims 175 to 179, wherein the second time period is about 6 weeks to about 12 weeks.

181. The composition according to claim 180, wherein the second time period is about 8 weeks.

182. The composition according to claim 180, wherein the second time period is about 12 weeks.

183. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, the method comprising administering to the subject a first dose and a second dose of the composition; The first dose comprises about 2 mg / kg of the composition administered once every four weeks during a first time period; and The second dose comprises about 2 mg / kg of the composition administered (i) once every two weeks or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period, wherein the anti-TNFα antibody comprises: (i) a first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) a second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) a third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) a fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) a fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) a sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) a seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) The twelfth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

184. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of treating or reducing sarcoidosis in a subject in need thereof, the method comprising administering a first dose and a second dose of the composition to the subject; The first dose comprises about 2 mg / kg of the composition administered bi-weekly during a first time period; and The second dose comprises (i) about 2 mg / kg of the composition administered bi-weekly during a second time period, or (iii) about 4 mg / kg of the composition administered once every 4 weeks, wherein the anti-TNFα antibody comprises: (i) A first CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:3; (ii) A second CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:4; (iii) A third CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:5; (iv) A fourth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:6; (v) A fifth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:7; (vi) A sixth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:8; (vii) A seventh CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:10; (viii) An eighth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) A ninth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) A tenth CDR, which has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) The eleventh CDR, wherein the eleventh CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 14; and (xii) The twelfth CDR, wherein the twelfth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:

15.

185. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of the composition; The first dose comprises about 2 mg / kg of the composition administered once every four weeks during a first time period; and The second dose comprises about 2 mg / kg of the composition administered (i) once every two weeks or (iii) about 4 mg / kg of the composition administered once every 4 weeks during a second time period, wherein the anti-TNFα antibody comprises: (i) A first CDR, wherein the first CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 3; (ii) A second CDR, wherein the second CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 4; (iii) A third CDR, wherein the third CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 5; (iv) A fourth CDR, wherein the fourth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 6; (v) A fifth CDR, wherein the fifth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 7; (vi) A sixth CDR, wherein the sixth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 8; (vii) A seventh CDR, wherein the seventh CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 10; (viii) An eighth CDR, wherein the eighth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to YA; (ix) A ninth CDR, wherein the ninth CDR has an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO:

15.

186. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of the composition; the first dose comprising about 2 mg / kg of the composition administered once every four weeks during a first time period; and the second dose comprising (i) about 2 mg / kg of the composition administered once every two weeks, or (iii) about 4 mg / kg of the composition administered once every 4 weeks, during a second time period, wherein the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to SEQ ID NO: 10; (viii) An eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99%, or 100% identical to YA; (ix) A ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 12; (x) A tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 13; (xi) An eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 14; and (xii) A twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

187. A composition comprising an anti-TNFα antibody sialylated with one or more N-acetylneuraminic acid molecules, the composition being used in a method of reducing granuloma size or reducing granuloma formation in a subject in need thereof relative to a control, the method comprising administering to the subject a first dose and a second dose of the composition; the first dose comprising about 2 mg / kg of the composition administered biweekly for a first period of time; and the second dose comprising (i) about 2 mg / kg of the composition administered biweekly for a second period of time, or (iii) about 4 mg / kg of the composition administered once every 4 weeks, wherein the anti-TNFα antibody comprises: (i) A first CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 3; (ii) A second CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 4; (iii) A third CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 5; (iv) A fourth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 6; (v) A fifth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 7; (vi) A sixth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 8; (vii) A seventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO: 10; (viii) an eighth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to YA; (ix) a ninth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:12; (x) a tenth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:13; (xi) an eleventh CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:14; and (xii) a twelfth CDR having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

15.

188. The composition according to any one of claims 160 to 187, further comprising administering (c) a third dose of about 4 mg / kg of the composition approximately once every two weeks for a third time period.

189. The composition according to claim 187, wherein the third time period is at least about 24 weeks.

190. The composition according to any one of claims 140 to 189, wherein the anti-TNFα antibody comprises a variable heavy (VH) domain having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

16.

191. The composition according to any one of claims 140 to 189, wherein the anti-TNFα antibody comprises a variable light (VL) domain having an amino acid sequence that is at least about 80%, 85%, 90%, 95%, 99% or 100% identical to SEQ ID NO:

9.

192. The composition according to any one of claims 140 to 191, wherein the anti-TNFα antibody comprises an asparagine-linked glycosylation site located on at least one heavy chain within the constant Fc region.

193. The composition according to claim 192, wherein the anti-TNFα antibody comprises asparagine-linked glycosylation sites located on both heavy chains within the constant Fc region.

194. The composition according to c, wherein the anti-TNFα antibody is sialylated at Asn 297.

195. The composition according to any one of claims 140 to 194, wherein the anti-TNFα antibody does not contain Neu5Gc.

196. The composition according to any one of claims 140 to 195, wherein the anti-TNFα antibody is an IgG antibody.

197. The composition according to claim 196, wherein the IgG antibody is an IgG1 antibody.

198. The composition according to claim 197, wherein the IgG1 antibody comprises a κ isotype.

199. The composition according to any one of claims 140 to 198, wherein the anti-TNFα antibody is a chimeric antibody.

200. The composition according to any one of claims 140 to 199, wherein the anti-TNFα antibody comprises a C-terminal lysine on at least one heavy chain.

201. The composition according to claim 200, wherein the anti-TNFα antibody comprises C-terminal lysines on both heavy chains.

202. The composition according to claim 200 or 201, wherein the C-terminal lysine is not trimmed.

203. The composition according to any one of claims 140 to 202, wherein the composition is administered by infusion.

204. The composition according to claim 203, wherein the infusion is an intravenous (IV) infusion.

205. The composition according to claim 203 or 204, wherein the infusion is administered over a period of at least about 2 hours.

206. The composition according to claim 203 or 204, wherein the infusion is administered over a period of about 2 hours.

207. The composition according to any one of claims 140 to 206, wherein the subject receives a cumulative dose of about 6 mg / kg of the composition during the first period.

208. The composition according to any one of claims 140 to 206, wherein the subject receives a cumulative dose of about 8 mg / kg of the composition during the first period.

209. The composition according to any one of claims 140 to 206, wherein the subject receives a cumulative dose of about 16 mg / kg of the composition during the first period.

210. The composition according to any one of claims 181 to 209, wherein the subject receives a cumulative dose of about 10 mg / kg to about 16 mg / kg of the composition during the second period.

211. The composition according to claim 210, wherein the subject receives a cumulative dose of about 10 mg / kg of the composition during the second period.

212. The composition according to claim 210, wherein the subject receives a cumulative dose of about 12 mg / kg of the composition during the second period.

213. The composition according to claim 210, wherein the subject receives a cumulative dose of about 14 mg / kg of the composition during the second period.

214. The composition according to claim 210, wherein the subject receives a cumulative dose of about 16 mg / kg of the composition during the second period.

215. The composition according to any one of claims 188 to 214, wherein the subject receives a cumulative dose of about 28 mg / kg of the composition.

216. The composition according to any one of claims 140 to 215, wherein the subject is administered a cumulative dose of from about 30 mg / kg to about 60 mg / kg of the composition.

217. The composition according to claim 216, wherein the subject is administered a cumulative dose of about 46 mg / kg of the composition.

218. The composition according to claim 216, wherein the subject is administered a cumulative dose of about 48 mg / kg of the composition.

219. The composition according to claim 216, wherein the subject is administered a cumulative dose of about 50 mg / kg of the composition.

220. The composition according to claim 216, wherein the subject is administered a cumulative dose of about 52 mg / kg of the composition.

221. The composition according to any one of claims 140 to 220, wherein the subject is administered at least 3 doses of the composition.

222. The composition according to any one of claims 140 to 221, wherein the subject is administered at least 4 doses of the composition.

223. The composition according to any one of claims 140 to 222, wherein the subject is administered at least 6 doses of the composition.

224. The composition according to any one of claims 140 to 223, wherein the subject is administered at least 7 doses of the composition.

225. The composition according to claim 221, wherein the at least 3 doses are administered over a 4-week period.

226. The composition according to claim 221, wherein the at least 3 doses are administered over an 8-week period.

227. The composition according to claim 221, wherein the at least 3 doses are administered over a 12-week period.

228. The composition according to claim 222, wherein the at least 4 doses are administered over a 9-week period.

229. The composition according to claim 222, wherein the at least 4 doses are administered over a 10-week period.

230. The composition according to claim 222, wherein the at least 4 doses are administered over an 11-week period.

231. The composition according to claim 222, wherein the at least 4 doses are administered over a 12-week period.

232. The composition according to claim 223, wherein the at least 6 doses are administered over a 10-week period.

233. The composition according to claim 223, wherein the at least 6 doses are administered over a 13-week period.

234. The composition according to claim 223, wherein the at least 6 doses are administered over a 16-week period.

235. The composition according to claim 223, wherein the at least 6 doses are administered over a 17-week period.

236. The composition according to claim 223, wherein the at least 6 doses are administered over a 20-week period.

237. The composition according to claim 234, wherein the at least 7 doses are administered over a 12-week period.

238. The composition according to claim 234, wherein the at least 7 doses are administered over a 15-week period.

239. The composition according to claim 234, wherein the at least 7 doses are administered over a 21-week period.

240. The composition according to claim 234, wherein the at least 7 doses are administered over a 24-week period.

241. The composition according to claim 234, wherein the at least 7 doses are administered over a 28-week period.

242. The composition according to any one of claims 140 to 241, wherein administration of the composition reduces the level of one or more transcripts selected from the group consisting of angiotensin converting enzyme 2 (ACE2), interleukin 6 (IL6), interleukin 2 receptor (IL2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL1B), interleukin 12 (IL12), interleukin 18 (IL18), interferon γ (IFNG), interleukin 8 (IL8), monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), chemokine ligand 2 (CXCL2), chemokine ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JunB), mitogen-activated protein kinase (MKP1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL32), intercellular adhesion molecule 1 (ICAM-1), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), and vascular cell adhesion molecule 1 (VCAM-1) relative to a control.

243. The composition according to claim 242, wherein the level of the one or more transcripts is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

244. The composition according to any one of claims 140 to 243, wherein the administration of the composition reduces the level of one or more proteins selected from the group consisting of angiotensin-converting enzyme 2 (ACE2), interleukin 6 (IL-6), interleukin 2 receptor (IL-2R), tumor necrosis factor α (TNFα), C-reactive protein (CRP), interleukin 1B (IL-1B), interferon γ (IFN-γ), interleukin 8 (IL-8), interleukin 12 (IL-12), interleukin 18 (IL-18), monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), chemokine ligand 2 (CXCL2), chemokine ligand 3 (CXCL3), early growth response 1 (EGR1), JunB proto-oncogene (JUNB), mitogen-activated protein kinase phosphatase 1 (MKP-1), TNFα-induced protein 1 (TNFAIP1), TNFα-induced protein 2 (TNFAIP2), TNFα-induced protein 3 (TNFAIP3), syndecan-4 (SDC4), superoxide dismutase 2 (SOD2), cytochrome c oxidase subunit II (COX2), interleukin 32 (IL-32), Krebs von den Lungen-6 (KL-6), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), chitinase 3-like 1 (CHIT1), secretoglobin family 1A member 1 (SCGB1A1), serum amyloid A1 (SAA1), intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1) relative to a control.

245. The composition according to claim 244, wherein the level of the one or more proteins is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

246. The composition according to any one of claims 140 to 245, wherein the administration of the composition increases one or more of the following relative to a control: forced vital capacity (FVC) measurement, forced expiratory volume in 1 second (FEV1) measurement, Leicester Cough Questionnaire (LCQ) score, Short Form 36 Health Survey (SF-36) score, King's Sarcoidosis Questionnaire (KSQ) score, Steroid Toxicity Scale (STS) score, or 6-minute walk test (6MWT) distance.

247. The composition according to claim 246, wherein the FVC measurement is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to the control.

248. The composition according to claim 246, wherein the FEV1 measurement result is increased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

249. The composition according to claim 246, wherein the LCQ score is increased by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 points relative to the control.

250. The composition according to claim 246, wherein the SF-36 score is increased by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 points relative to the control.

251. The composition according to claim 246, wherein the KSQ score is increased by at least 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, 39 points, 40 points, 41 points, 42 points, 43 points, 44 points, 45 points, 46 points, 47 points, 48 points, 49 points, 50 points, 51 points, 52 points, 53 points, 54 points, 55 points, 56 points, 57 points, 58 points, 59 points, 60 points, 61 points, 62 points, 63 points, 64 points, 65 points, 66 points, 67 points, 68 points, 69 points, 70 points, 71 points, 72 points, 73 points, 74 points, 75 points, 76 points, 77 points, 78 points, 79 points, 80 points, 81 points, 82 points, 83 points, 84 points, 85 points, 86 points, 87 points, 88 points, 89 points, 90 points, 91 points, 92 points, 93 points, 94 points, 95 points, 96 points, 97 points, 98 points, 99 points or 100 points relative to the control.

252. The composition according to claim 246 or 251, wherein the KSQ score is the KSQ general health status module or the KSQ lung score.

253. The composition according to claim 246, wherein the STS score is increased by at least 1 point, 2 points, 3 points, 4 points, 5 points or 6 points relative to the control.

254. The composition according to claim 246, wherein the 6MWT distance is increased by at least 10 m, 20 m, 30 m, 40 m, 50 m, 60 m, 70 m, 80 m, 90 m, 100 m, 125 m, 150 m, 175 m, 200 m, 250 m, 300 m, 350 m, 400 m, 450 m or 500 m relative to the control.

255. The composition according to any one of claims 140 to 254, wherein the administration of the composition reduces one or more of the following relative to the control: the patient global assessment (PGA) score, the fatigue assessment scale (FAS) score, the modified medical research council (mMRC) dyspnea scale score, the St. George's respiratory questionnaire (SGRQ) score or the Borg CR10 dyspnea score.

256. The composition according to claim 255, wherein the PGA score is reduced by at least about 1 point, 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points or 10 points relative to the control.

257. The composition according to claim 255, wherein the FAS score is reduced by at least about 1 point, 2 points, 3 points or 4 points relative to the control.

258. The composition according to claim 255, wherein the mMRC scale score is reduced by at least about 1, 2, 3 or 4 levels relative to the control.

259. The composition according to claim 255, wherein the SGRQ score is reduced by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 points relative to the control.

260. The composition according to claim 255, wherein the Borg CR10 dyspnea score is reduced by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 points relative to the control.

261. The composition according to any one of claims 140 to 260, wherein administration of the composition reduces the need for anti-inflammatory drugs relative to a control.

262. The composition according to claim 261, wherein the anti-inflammatory drug is selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, ezogabine, rituximab, adalimumab, golimumab, namilumab and infliximab.

263. The composition according to claim 262, wherein the corticosteroid is selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone and triamcinolone.

264. The composition according to any one of claims 261 to 263, wherein the reduced need for anti-inflammatory drugs comprises a reduction in the frequency, dose or duration of administration of the anti-inflammatory drug.

265. A composition according to any one of claims 140 to 264, wherein administration of the composition reduces granuloma formation or decreases the granuloma size in the subject by about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to a control.

266. The composition according to claim 265, wherein the granuloma is a cutaneous granuloma, a pulmonary granuloma, a lymph node granuloma, an ocular granuloma, a hepatic granuloma, a splenic granuloma, a cardiac granuloma or a rheumatoid granuloma.

267. A composition according to any one of claims 140 to 266, wherein administration of the composition reduces the level of calcitriol in the subject relative to a control.

268. The composition according to claim 267, wherein the level of calcitriol is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

269. A composition according to any one of claims 140 to 268, wherein administration of the composition reduces the level of neopterin in the subject relative to a control.

270. The composition according to claim 260, wherein the level of neopterin is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

271. A composition according to any one of claims 140 to 270, wherein administration of the composition reduces the level of lysozyme in the subject relative to a control.

272. The composition according to claim 271, wherein the level of lysozyme is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

273. A composition according to any one of claims 140 to 272, wherein administration of the composition reduces the level of soluble IL-2R (sIL-2R) in the subject relative to a control.

274. The composition according to claim 273, wherein the level of sIL-2R is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the control.

275. A composition according to any one of claims 140 to 274, wherein the subject has one or more of the following: acute sarcoidosis, chronic sarcoidosis, progressive sarcoidosis, pulmonary sarcoidosis, remitting sarcoidosis, refractory sarcoidosis, advanced sarcoidosis, ocular sarcoidosis, cutaneous sarcoidosis or nervous system sarcoidosis.

276. A composition according to any one of claims 140 to 275, wherein the subject has received an anti-inflammatory drug.

277. The composition according to any one of claims 140 to 276, wherein the subject is receiving an anti-inflammatory drug.

278. The composition according to claim 276 or 277, wherein the anti-inflammatory drug is selected from the group consisting of: corticosteroids, methotrexate, azathioprine, pentoxifylline, thalidomide, leflunomide, mycophenolate mofetil, cyclophosphamide, chloroquine, repository corticotropin (RCI), hydroxychloroquine, etrolizumab, rituximab, adalimumab, golimumab, namilumab, and infliximab.

279. The composition according to claim 278, wherein the corticosteroid is selected from the group consisting of: prednisone, prednisolone, dexamethasone, hydrocortisone, methylprednisolone, betamethasone, cortisone, fludrocortisone, and triamcinolone.

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