CD40l antagonist and uses thereof

Abstract

To provide a CD40L antagonist and uses thereof.SOLUTION: The present invention provides a human CD40L-specific Tn3 molecule and therapeutic uses thereof. The description further provides a method for reducing a measure of RA disease activity in a patient being treated for RA. The method includes steps of administering VIB4920 to the patient and reducing the measure of RA disease activity in the patient. The measure of RA disease activity reduced may include one or more of DAS28-CRP, clinical disease activity index (CDAI), tender joint count, swollen joint count, patient's global assessment or physician's global assessment. The VIB4920 may be administered at a dose of between approximately 500 mg and 3000 mg.SELECTED DRAWING: None

Description

[Background technology]

[0001] The CD40 / CD40L pathway plays an important role in initiating humoral immune responses, It is involved in the pathogenesis of several autoimmune diseases. CD40 is expressed on dendritic cells (DCs), macrophages, and It is constitutively expressed on a variety of antigen-presenting cells, including hematopoietic and B cells (1), and on non-hematopoietic cells (2). It may also be expressed.

[0002] The CD40 ligand, CD40L (also known as CD154), is highly expressed It is regulated by the IL-1 receptor and is found predominantly on activated CD4+ T cells (2). CD40 / CD40L interactions between T cells mediate effective secretion of T cell-dependent antigens. The CD40 / CD40L system is essential for initiating immune responses (3-5). In vivo, it induces B cell proliferation, differentiation, and isotype switching (6-9). It is known that GC formation, somatic hypermutation, and memory B cell and long-term survival characteristics are CD40 signaling is required for the production of inflammatory cells (10-13). Deficiency of CD40 or CD40L results in a disorder characterized by impaired isotype class switching. This results in X-linked hyperimmunoglobulin syndrome, which is difficult to detect. High levels of serum IgG, IgA, or IgE with increased susceptibility to infection It appears as IgM ( 14 – 16 ).

[0003] Clinical trials with compounds directed against CD40L have demonstrated that CD40L inhibits the expression of CD40 in autoimmune diseases. The potential of targeting 40 pathways has been demonstrated. In a Phase 2 study, humanized 5c 8Anti-CD40L antibody BG9588 has been shown to reduce proteinuria and anti-CD40L in patients with proliferative lupus nephritis. Anti-CD40L treatment significantly reduced the titer of dsDNA antibodies to IgG1. Circulating CD38 in SLE patients hi Decreased Ig-secreting cells and peripheral GC B cells Further studies have revealed that anti-CD40 Treatment with L monoclonal antibody (mAb) in patients with immune thrombocytopenia (ITP) It has been shown that a significant response is induced in a subset of patients ( 20 ).

[0004] Anti-CD40L mAb treatment shows promise in clinical trials Nevertheless, these programs have been discontinued due to adverse thromboembolic events. Although not precisely defined, one possible explanation for these unexpected safety issues is One explanation is that FcγR on platelets is expressed in humans but not in mice. IIa (or CD32a) expression (21). CD40L is expressed on activated platelets. are also highly expressed (22) and correlate with both CD40L and FcγRIIa on adjacent cells. Simultaneous antibody-mediated binding to the antibody may induce platelet aggregation. The role of FcγRIIa in anti-CD40L-induced thrombocytopenia was confirmed by In transgenic mice that have been introduced with human FcγRIIa, anti-CD40 LmAb induced shock and thrombocytopenia (23). This effect was consistent with the Wild-type mice or aglycosylated versions of the antibodies that are unable to associate with cγR No adverse events were observed in any of the transgenic mice injected with nicotinamide.

[0005] Targeting CD40L but without the potential complications associated with mAbs To achieve this, a CD40L-specific Tn3 scaffold protein (24, 25) was generated. The protein is derived from the third fibronectin type III domain of human tenascin-C and can be engineered to confer target-specific binding properties (26, 27). Fusing the bivalent CD40L-specific Tn3 protein to human serum albumin (HSA) and a molecule capable of binding to human CD40L and blocking its interaction with the CD40 receptor. , i.e., VIB4920, was obtained. Consistent with this inhibition of the CD40L / CD40 interaction, VIB4920 inhibits in vivo CD40 signaling events by blocking them. Toro was able to potently inhibit the activation and differentiation of human B cells.

[0006] Novel therapeutic approaches to significantly affect humoral immune responses and treat autoimmune and / or inflammatory conditions There is a need in the art for new treatments. There is also a need to induce immunotolerance into replacement therapy for patients in need.

[0007] Currently, VIB4920 is administered to patients suffering from an autoimmune / inflammatory disease or disorder. It has been shown that serotonin-dependent steroids reduce clinical symptoms and other markers of disease. When administered at certain doses to subjects with rheumatoid arthritis (RA), VIB4920 produced rheumatoid Factor (RF) autoantibody titer, Vectra DA biomarker score, DAS28- A statistically significant reduction in disease activity as measured by CRP compared to placebo This results in: Summary of the Invention [Means for solving the problem]

[0008] Provided herein are methods for suppressing B cell and T cell mediated immune responses in a subject. The method provides a dose of 500 mg to 3000 mg of VIB4920 for patients requiring it. The method includes administering to a subject a therapeutically effective amount of the antibody to suppress B cell and T cell mediated immune responses.

[0009] Also provided herein is a method for treating an autoimmune disease or disorder. administering a dose of VIB4920 between 0.00 mg and 3,000 mg to a subject in need thereof; thereby treating an autoimmune disease or disorder.

[0010] The present specification describes a method for reducing indicators of RA disease activity in patients undergoing treatment for RA. The method further comprises administering VIB4920 to a patient and assessing RA disease activity in the patient. The method includes the step of reducing the DAS28-CR as an index of reduced RA disease activity. P, Clinical Disease Activity Index (CDAI), tender joint count, swollen joint count, patient global assessment, or a physician's global assessment. It may be administered in doses of 500mg to 3000mg.

[0011] The present specification also describes a method for reducing RF autoantibodies in patients undergoing RA treatment. The method comprises administering VIB4920 to a patient at a dose of about 500 mg to 3000 mg. The method includes reducing RF autoantibodies in a patient.

[0012] The present specification also provides a method for reducing biomarker scores in patients undergoing RA treatment. The method comprises administering to a patient about 500 mg to 3000 mg of VIB4920. and decreasing the biomarker score in the patient. The biomarker score is a combination of the plasma cell (PC) gene signature, Vectra- The clinical features of the present invention may be one or more of the following: DA score, serum C-reactive protein (CRP) level, stomach.

[0013] The present disclosure also provides a method for reducing a PC gene signature score in a patient in need thereof. The method comprises administering VIB4920 to a patient in need thereof; The method includes the step of decreasing the PC gene signature score of the patient. are systemic lupus erythematosus, rheumatoid arthritis, myositis, antiphospholipid syndrome, autoimmune Hepatitis, Sjogren's disease, or other autoimmune or inflammatory conditions, as well as transplant surgery and graft-recipient The patient may be undergoing treatment for a host disease. may be in a dose of about 500 mg to 3000 mg.

[0014] The present disclosure relates to the treatment of autoantibodies in patients undergoing treatment for autoimmune disorders, or alloantibodies in the case of transplantation. The present invention further provides a method for reducing the amount of and administering the compound to a patient to reduce autoantibodies or alloantibodies in the patient. In the present invention, the patient is a patient undergoing treatment for an autoimmune disease characterized by the presence of autoantibodies. or the patient is a patient undergoing treatment to prevent transplant rejection. Patients will receive VIB4920 at doses ranging from approximately 500 mg to 3000 mg.

[0015] Also provided herein is a method for reducing inflammation in a patient. 20 to a patient in need thereof, thereby reducing inflammation in the patient. The patient may be undergoing treatment for an inflammatory disease or disorder, or may be undergoing organ or tissue transplantation. The patient may be receiving prophylactic treatment for anticipated inflammation in response to the 920 may be administered in doses of approximately 500 mg to 3000 mg.

[0016] Further provided herein is a method of inducing immune tolerance to a replacement therapy in a patient. The method comprises administering VIB4920 to a patient in need of replacement therapy, and inducing replacement therapy in the patient. The step of reducing immune tolerance to VIB4920 is about 1000 mg to 3000 mg. It may be administered in doses of mg. [Brief description of the drawings]

[0017] [Fig. 1A-1G]

[0023] Figure 1 provides the biochemical properties of human CD40L-specific Tn3 clones, including the biochemical properties of VIB4920, a bivalent 342 clone fused to HSA. Figure 1A shows the ability of a series of human CD40L-specific Tn3 clones to inhibit CD40-CD40L interaction as measured by Proteon. Percent inhibition is shown across a range of Tn3 concentrations. The average of duplicate wells is shown. Figure 1B shows the ability of anti-CD40L Tn3 protein to inhibit CD40L-mediated signaling through NFkB. HEK293 cells expressing CD40R and an NFkB-luciferase reporter were stimulated with recombinant CD40L overnight in the presence of anti-CD40L Tn3 protein. Percent inhibition of luciferase activity is shown. Data represent the average of duplicate wells. Figure 1C shows the inhibition of CD86 upregulation by Tn3 constructs at various concentrations. Human PBMCs were stimulated with recombinant CD40L and preincubated with CD40L Tn3 protein, after which CD86 expression was assessed by flow cytometry. Means of duplicate wells are shown. In Figure 1D, the indicated Tn3 molecules were examined in an ELISA assay for their ability to inhibit the CD40-CD40L interaction. Data represent the mean of duplicate wells. In Figure 1E, data are provided from screening of clone 342 for binding to a panel of related TNF family members, including Fas, TNFα, TNFβ, and OX40L. Clone 342 was found to selectively bind CD40L. In Figure 1F, the proposed structure of VIB4920 is based on the crystallization of 342 Tn3 and the published crystal structure of HSA (1). In Figure 1G, a schematic of the CD40 / CD40L and 342 / CD40L structures arranged via a common CD40L molecule is shown. CD40L is shown in green, Tn3 in magenta, and the CD40 receptor in cyan. [Figures 2A-2D]Figure 2 provides structural features of human CD40L-specific Tn3 clone 342. Figure 2A shows a schematic of the trimeric 342 / CD40L structure. The extracellular domain of CD40L is shown in green and 342 in magenta. Figure 2B shows the interface between 342 and CD40L. Fragments of CD40L and 342 are shown as green and magenta tubes, respectively. Amino acids involved in hydrogen bonds are shown as sticks. Hydrogen bonds are shown as dashed black lines with associated distances (Å). Bonds with distances up to 3.5 Å are shown. Figures 2C and 2D illustrate the electrostatic surface potential of the surface interactions of CD40L (in Figure 2C) and 342 CD40L-specific Tn3 (in Figure 2D). To show the interface, the molecule has been rotated approximately 90 degrees and made semi-transparent to allow visualization of the amino acids involved in hydrogen bonds. Red indicates negatively charged surfaces and blue indicates positive charges. [Figure 3A-3G]Figure 3 shows how VIB4920 inhibits CD40 signaling and human B cell activation but does not induce platelet aggregation in ex vivo studies. Figure 3A shows the percentage of inhibition of NFkB luciferase signal by VIB4920 in modified HEK29 cells stimulated overnight with CD40L. Data represent the mean of duplicate wells. One of two independent experiments is shown. Figure 3B shows how VIB4920 and anti-CD40L mAb were able to inhibit the upregulation of CD86 in stimulated human PBMC. Human PBMC were stimulated with recombinant human megaCD40L for 24 hours and the percentage of CD19+ / CD86+ cells was measured by flow cytometry. Data represent the mean of duplicate wells. Figure 3C shows that human B cells were stimulated with IL-21 and megaCD40L in the presence of control or anti-CD40L (mAb or VIB4920 Tn3). B cell proliferation was quantified on day 3. The dotted line represents ATP levels in unstimulated cells. Data shown are the mean and SD of triplicate wells and are representative of two independent experiments. Figures 3D and 3E show the effect of anti-CD40L (mAb or VIB4920 Tn3) on human B cells left unstimulated (none) or stimulated with IL-21, anti-IgM and megaCD40L. The number of PCs was quantified on day 7. Figure 3D illustrates the effect of the indicated molecules on day 7 on the percentage of IgD-CD38hi PCs on day 7. Figure 3E shows the effect of the indicated molecules at the indicated concentrations on the number of PCs on day 7. Data shown are the mean and SD of triplicate wells and are representative of two independent experiments. ****=p<0.0001 by two-tailed unpaired Student's t-test. Figures 3F and 3G show the effect of anti-CD40L (mAb or VIB4920 Tn3) molecules on washed human platelets incubated with preformed immune complexes, and platelet aggregation or lack thereof was measured for 12-14 min. Figure 3F shows the aggregation rate. Shown here are platelets preincubated with anti-CD32a antibody for 5 min before adding immune complexes. Adenosine diphosphate (ADP) was used as a positive control for aggregation.Figure 3G provides the percentage of aggregation following incubation of platelets with immune complexes of VIB4920 (Tn3) or anti-CD40L mAb (5c8) at the indicated concentrations. Data are representative of two independent experiments. [Figure 4A-4B] Mouse surrogate CD40L-specific Tn3 responds to immunization and shows potent neutralizing activity in vivo. In both Figures 4A and 4B, mice were immunized with sheep red blood cells (SRBC) on day 0 and control or anti-CD40L Tn3 (M31-MSA) was administered daily for days 9-13. Figure 4A provides the percentage of germinal center B cells in spleen and lymph nodes quantified by flow cytometry on day 14. Points indicate individual animals and data are representative of two independent experiments. ****=p<0.0001 by two-tailed unpaired Student's t-test. Figure 4B provides the production of anti-sheep red blood cell IgG quantified from serum on day 14. Data show the mean and SEM of 4 animals per group (n=1 study). [Figure 5A-5B] Figure 5A is a study design for a Phase 1a clinical trial to evaluate the safety of VIB4920 in healthy volunteers. Figure 5A shows the study cohorts for the Phase 1a study. Figure 5B provides the dosing and immunization regimen for the Phase 1a study. [Figure 6A-6B] In a Phase 1a study in healthy volunteers, VIB4920 demonstrated a favorable PK profile. Figure 6A shows blood concentrations of VIB4920 measured by ELISA at the indicated time points. The dotted line represents the lower limit of sensitivity of the assay. Error bars represent standard deviation of the mean but were not calculated for groups of N=2 subjects. Figure 6B shows concentrations of soluble CD40L assessed by ELISA in all dose cohorts at the indicated time points. The dotted line represents the lower limit of detection of the assay. [Figure 7]VIB4920 inhibits anti-drug antibodies (ADA) at high doses in a Phase 1a study in healthy volunteers. Presence of ADA was measured by ELISA. Each subject within each cohort is represented by a separate line. Subjects with high ADA (>median titer 480) are shown by a magenta line, subjects with low ADA (<median titer 480) are shown by a dark blue line, and subjects with undetectable ADA are highlighted in light blue. [Figure 8A-8C] VIB4920 inhibits B cell proliferation and TDAR in a dose-dependent manner in healthy human volunteers. Healthy volunteers were immunized with KLH 14 days prior to treatment with placebo or VIB4920 and rechallenged 15 days after administration. Figure 8A provides anti-KLH IgG titers in healthy volunteers at different VIB4920 doses over multiple time points. Figure 8B provides anti-KLH IgM titers in healthy volunteers at different VIB4920 doses over multiple time points. IgG and IgM titers were measured by ELISA. Figure 8C is a dose-response model for inhibition of anti-KLH IgG at day 43. [Figure 9A-9C]VIB4920 inhibits B cell proliferation and plasma cell responses upon TDAR reduction in healthy human subjects. Figure 9A provides the detected frequency of circulating proliferating B cells (Ki67+ CD19+) quantified by flow cytometry at various time points in volunteers who received either placebo or high dose VIB4920 in the TDAR validation study. Figure 9B provides the detected frequency of circulating class-switched memory B cells (Ki67+ CD19+ IgD-CD27+) quantified by flow cytometry at various time points in volunteers who received either placebo or high dose VIB4920 in the TDAR validation study. Figure 9C provides the PC signature score in whole blood assessed by Taqman PCR. Mean and standard error expression values ​​for placebo and high dose VIB4920 groups are shown. *=P<0.05, **=P<0.01 compared to placebo by Mann-Whitney U test. [Figure 10] FIG. 1B is a Phase 1b study design for evaluating VIB4920 in patients with RA. Arrows indicate administration of VIB4920 or placebo. [Figure 11] Demographic and clinical characteristics of the cohort of RA patients in a phase 1b clinical trial of VIB4920. [Figure 12] Figure 1 shows an acceptable safety profile in RA patients with VIB4920. The most common TEAEs occurring in at least two subjects in a Phase 1b study in RA subjects are shown. [Figures 13A-13C]Figure 13 shows linear PK and dose-dependent reduction of ADA in a Phase 1b study of RA patients with VIB4920. Figure 13A provides the concentration of VIB4920 in the circulating blood as measured by ELISA at the indicated time points. The dotted line represents the lower limit of sensitivity of the assay. The mean and standard error of the mean are shown. Figure 13B provides the percentage of subjects with positive ADA titers as measured by ELISA for each dose administered at any time point during the study. Figure 13C provides ADA titers over time in subjects with detectable ADA as measured by ELISA. [Fig. 14A-14F] VIB4920 reduces disease index scores and autoantibodies in RA patients. Figure 14A shows the change from baseline in DAS28-CRP (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 14B shows the change from baseline in CDAI (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 14C shows the change from baseline in Patient Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 14D shows the change from baseline in Physician Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 14E shows the change from baseline in Vectra DA score (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. FIG. 14F shows measurements of percent reduction in RF antibody titers, as measured by ELISA, at the indicated time points for VIB4920 or placebo at the indicated doses, respectively. [Figure 15A-15B]VIB4920 reduces DAS28-CRP scores and RF autoantibodies in RA patients in a dose-dependent manner. Figure 15A shows the difference in DAS28-CRP scores between placebo and the indicated doses of VIB4920 at day 85. A linear dose-response is shown and identified as the best model for evaluating the relationship between VIB4920 dose and reduction in disease activity. Figure 15B shows the percent reduction in RF autoantibodies at the indicated doses of VIB4920 compared to placebo at day 85. The Emax model is shown and determined to be the best model for evaluating the relationship of VIB4920 dose to RF titer. [Figure 16] VIB4920 improves the DAS28 category in treated RA patients. DAS28 category at day 85 is shown. At day 85, 50% and 75% of treated RA patients in the 1000 mg and 1500 mg groups, respectively, had low disease activity or were in remission. [Figures 17A-17C] The effect of VIB4920 on tender / swollen joint counts and CRP in a Phase 1b study of RA subjects. Figure 17A shows the change from baseline in tender joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 17B shows the change from baseline in swollen joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 17C shows the percentage change in CRP levels to baseline in RA subjects at the indicated doses and at the indicated time points. Means and standard errors are shown, respectively. [Figure 18] Amino acid sequence of the VIB4920 molecule. [Figure 19A-19B] Amino acid sequence of the clone 342 CD40L-specific Tn3 molecule. [Figure 20] Amino acid sequence of the bivalent clone 342 CD40L-specific Tn3 molecule. [Figure 21A-21B] Amino acid sequence of the clone 309 CD40L-specific Tn3 molecule. [Fig. 22A-22B]VIB4920 reduces disease index scores and autoantibodies in RA patients during both the 12-week VIB4920 administration period and the 12-week observation period after the last VIB4920 dose. Figure 22A shows the change from baseline in DAS28-CRP (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22B shows the change from baseline in CDAI (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22C shows the change from baseline in Patient Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22D shows the change from baseline in Physician Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22E shows the change from baseline in Vectra DA scores (mean and standard error shown) assessed at the indicated time points for specific doses of VIB4920 or placebo. Figure 22F shows measurements of the percent reduction in RF antibody titers measured by ELISA at the indicated time points for the indicated doses of VIB4920 or placebo, respectively. [Fig. 22C-22D]VIB4920 reduces disease index scores and autoantibodies in RA patients during both the 12-week VIB4920 administration period and the 12-week observation period after the last VIB4920 dose. Figure 22A shows the change from baseline in DAS28-CRP (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22B shows the change from baseline in CDAI (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22C shows the change from baseline in Patient Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22D shows the change from baseline in Physician Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22E shows the change from baseline in Vectra DA scores (mean and standard error shown) assessed at the indicated time points for specific doses of VIB4920 or placebo. Figure 22F shows measurements of the percent reduction in RF antibody titers measured by ELISA at the indicated time points for the indicated doses of VIB4920 or placebo, respectively. [Fig. 22E-22F]VIB4920 reduces disease index scores and autoantibodies in RA patients during both the 12-week VIB4920 administration period and the 12-week observation period after the last VIB4920 dose. Figure 22A shows the change from baseline in DAS28-CRP (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22B shows the change from baseline in CDAI (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22C shows the change from baseline in Patient Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22D shows the change from baseline in Physician Global Assessment (mean and standard error shown) assessed at the indicated time points for a particular dose of VIB4920 or placebo. Figure 22E shows the change from baseline in Vectra DA scores (mean and standard error shown) assessed at the indicated time points for specific doses of VIB4920 or placebo. Figure 22F shows measurements of the percent reduction in RF antibody titers measured by ELISA at the indicated time points for the indicated doses of VIB4920 or placebo, respectively. [Figure 23A-23B] VIB4920 affects tender / swollen joint counts and CRP in RA patients. The effect of VIB4920 can be detected in a Phase 1b clinical trial in RA patients, both during the administration phase and during the 12-week observation period after administration. Figure 23A shows the change from baseline in tender joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 23B shows the change from baseline in swollen joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 23C shows the change in the ratio of CRP levels to baseline in RA subjects at the indicated doses and at the indicated time points. Mean and standard error are shown, respectively. [Figure 23C]VIB4920 affects tender / swollen joint counts and CRP in RA patients. The effect of VIB4920 can be detected in a Phase 1b clinical trial in RA patients, both during the administration phase and during the 12-week observation period after administration. Figure 23A shows the change from baseline in tender joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 23B shows the change from baseline in swollen joint counts in RA subjects at the indicated doses and at the indicated time points. Figure 23C shows the change in the ratio of CRP levels to baseline in RA subjects at the indicated doses and at the indicated time points. Mean and standard error are shown, respectively. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] Methods for suppressing B cell mediated immune responses, methods for treating autoimmune diseases or disorders, inflammation Methods for reducing autoantibodies in a patient, methods for reducing autoantibodies in a patient, and indicators of RA disease activity in a patient. Methods for reducing RF autoantibodies in patients, methods for reducing RF autoantibodies in patients, genetic sequencing of plasma cells in patients Methods for reducing tumor score and inducing immune tolerance to replacement therapy in patients - Patents.com Described herein is VIB4920 and its utility in methods for treating

[0019] When VIB4920 is used to treat an autoimmune disease or disorder, VIB49 20 is for alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, and Autoimmune diseases of the kidney, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and testicular inflammation, Sjogren's syndrome, psoriasis, atherosclerosis, diabetic retinopathy and other retinopathies, Retrolental fibroplasia, age-related macular degeneration, neovascular glaucoma, hemangioma, thyroid hyperplasia (Graves corneal and other tissue transplants; and chronic inflammation, sepsis, rheumatoid arthritis, peritonitis, Crohn's disease, reperfusion injury, sepsis, endotoxic shock, cystic fibrosis, endocarditis, psoriasis, Arthritis (e.g., psoriatic arthritis), anaphylactic shock, organ ischemia and reperfusion injury, spinal cord injury Spinal cord injury and allograft rejection, autoimmune thrombocytopenia, Behçet's disease, bullous pemphigoid Smallpox, cardiomyopathy, celiac sprue dermatitis, chronic fatigue immune deficiency syndrome (CFIDS) ), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CRE ST syndrome, cold agglutinin disease, Crohn's disease, discoid lupus, essential mixed cryoglobulin anemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre, graft-versus-host syndrome Thyroiditis, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy , juvenile arthritis, lichen planus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, Ig G4-mediated diseases, multiple sclerosis, type 1 or immune-mediated diabetes, myasthenia gravis, pemphigus vulgaris Acne, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica , polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis Arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, stiff-man syndrome, systemic lupus erythematosus, Takayasu's arteritis, Cephalic arteritis / giant cell arteritis, ulcerative colitis, uveitis, ANCA-associated vasculitis, peeling herpetiformis Other vasculitis such as dermatomyositis, vitiligo, solid organ transplant rejection, graft-versus-host disease, kidney transplants Desensitization of panel-reactive antibodies in recipients, islet cell transplantation and allogeneic hematopoietic stem cell transplantation , focal segmental glomerulosclerosis (FSGS), and glomerulonephritis.

[0020] More specifically, VIB4920 is intended to be used in the treatment of RA, systemic lupus erythematosus (SLE), myositis, Antiphospholipid syndrome, autoimmune hepatitis, focal segmental glomerulosclerosis (FSGS), loop nephritis, inflammatory myopathy, idiopathic thrombocytopenic purpura (ITP), systemic sclerosis, vasculitis, Cutaneous lupus, autoimmune hemolytic anemia, myasthenia gravis, IgG4-related disease, or Sjogren's disease VIB4920 may also be used to treat Renn's syndrome. and / or to reduce or prevent organ or tissue transplant rejection It can be used.

[0021] Treatment of an autoimmune disease or disorder involves suppressing the B-cell or T-cell mediated immune response. This may be by reducing class-switched antibodies, circulating B cell subpopulations, Reducing the set, reducing plasma activity, or plasma cell and plasma cell genes The treatment of an autoimmune disease or disorder may involve reducing the inflammatory process. The inflammatory markers may be autoantibody levels, plasma cell ( PC) or PC gene signature (genes IGHA1, IGJ, IGKC, IGKV4 -1 and TNFRSF17 expression), circulating B cell subsets and and class-switched antibodies. Treatment is defined as the reduction of clinical signs and symptoms as measured by a patient or physician global assessment. Clinical signs and symptoms include arthritis, pain, fatigue, fever, fatigue, rash, weakness, or signs of organ failure such as protein in the urine or loss of kidney function You can name one or more of them.

[0022] If the method is to reduce autoantibodies in a patient undergoing treatment for an autoimmune disorder, The antibodies can be used, for example, to treat SLE, Sjogren's syndrome, inflammatory myopathies, or systemic sclerosis. The antinuclear antibodies may be anti-SSA / Ro autoantibodies or anti-SSB- La autoantibody (SLE or Sjogren's syndrome), anti-dsDNA antibody (SLE), anti-Smithk Antibodies to anti-topoisomerase (systemic sclerosis), or anti-histone antibodies (S The method may comprise administering to a patient undergoing treatment for an autoimmune disorder one or more of the following: In the case of reducing antibodies, autoantibodies are, for example, in patients undergoing treatment for autoimmune hepatitis. The method may be for treating an autoimmune disorder in a patient receiving treatment for an autoimmune disorder. In the case of reducing autoantibodies, the autoantibodies may be, for example, in patients undergoing treatment for myasthenia gravis. The antibody may be an anti-nicotinic acetylcholine receptor antibody, or an anti-muscle specific kinase antibody. If the method is to reduce antibodies in a patient undergoing transplant surgery, the antibodies are alloantibodies. That's fine.

[0023] Reducing autoantibodies in patients undergoing treatment for autoimmune disorders is Even if it reduces the proportion of autoantibodies to a level at least 20% lower than before administration, This is at least 10% lower than the autoantibody levels before treatment with VIB4920. At least 30%, at least 40%, at least 50%, at least 60%, at least 70% %, or at least to a level of 80%. Reduction of autoantibodies can be achieved within 1 to 3 months of initiating VIB4920 administration.

[0024] When the autoimmune disease or disorder is RA, treatment of rheumatoid arthritis includes treatment with RF autoantibodies, anti Citrullinated peptide antibody, Vectra DA biomarker score (interleukin 1 (IL1) α-6, tumor necrosis factor receptor type I, vascular cell adhesion molecule 1, epidermal growth factor, vascular endothelial cell adhesion molecule Growth factor A, YKL-40, matrix metalloproteinase 1, MMP-3, CRP, blood Vector, a comprehensive score of serum amyloid A, leptin, and resistin expression levels a DA biomarker score), plasma cell (PC) signature, and seroreactive C protein One of the following: CRP, DAS28-CRP, or Clinical Disease Activity Index (CDAI) or to reduce tender joint count, severity of swollen joint count, swollen joint count, or reducing the severity of swollen joints. If A, treatment is to achieve ACR20, ACR50, or ACR70. This is also fine.

[0025] Treating an autoimmune disease or disorder means: Reducing clinical symptoms of a disease or disorder by at least 20% or reducing inflammation or reducing a biomarker for a disease or disorder. A reduction in any of these symptoms, inflammation, or biomarkers may be At least 50% reduction in symptoms or inflammation compared to levels before treatment with 920 was started The reduction may be a reduction in a biomarker. The reduction may be an alleviation of the autoimmune disease or disorder. The cell may be characterized as being in a solution phase.

[0026] Furthermore, when the autoimmune disease or disorder is rheumatoid arthritis, the autoimmune disease or disorder is treated. The aim of this study is to determine whether patients' R-cell responses are improved by comparing their R-cell responses to RF autoantibodies before treatment with VIB4920. F autoantibodies, at least about 20%, at least 30%, at least 40%, at least 45%, at least 50%, at least 60%, at least 75%, or at least 80% % level. If the autoimmune disease or disorder is rheumatoid arthritis, In such cases, treating an autoimmune disease or disorder means decreasing DAS28-CRP. and decreasing DAS28-CRP may be at least -1.2, or An adjusted mean difference of at least -1.5, or at least -2.0, or at least -2.2 In addition, when the autoimmune disease or disorder is rheumatoid arthritis, In this case, treating an autoimmune disease or disorder means administering Vectra DA biomarker scores The reduction may be an adjusted mean difference of at least -10. 3, or at least -10.5, or at least -10.8.

[0027] When VIB4920 is used in a method for reducing inflammation, the inflammation can be an inflammatory disease. It may be the result of a disease or disorder, or may be the result of an organ or tissue transplant procedure. even if such damage is caused by or anticipates In a method for reducing inflammation in an inflammatory disease or disorder, When used, the inflammatory disease or disorder is an inflammatory myopathy, or lupus nephritis, cutaneous lupus erythematosus, or cutaneous lupus erythematosus. rheumatoid arthritis, RA, SLE, ITP, myositis, Sjogren's syndrome, vasculitis, systemic sclerosis, autoimmune diseases The condition may be autoimmune hemolytic anemia, myasthenia gravis, or focal segmental glomerulosclerosis. When VIB4920 is used in a method to reduce inflammation, the inflammation is an organ or tissue transplant. even if the injury is due to, or anticipated by, any injury resulting from the transplant procedure. It may also be something.

[0028] VIB4920 is used in a method for inducing immune tolerance to replacement therapy in patients In this case, VIB4920 may be able to reduce patients' production of neutralizing antibodies to the replacement therapy. If the patient is naïve to replacement therapy or if Otherwise, if the patient has not yet developed neutralizing antibodies to the replacement therapy, the first step is to develop immune tolerance. Induction of immune response prevents patients from producing neutralizing antibodies against the replacement therapy. However, if the patient develops neutralizing antibodies against the replacement therapy, VIB 4920 reduces the levels of neutralizing antibodies produced by patients against replacement therapy. By this, immune tolerance can be induced. The level of antibodies is at least 50%, at least 55%, at least 60%, at least 65%, at least 70, at least 75%, at least 80%, at least 85%, at least at least 90%, at least 95%, or to undetectable levels. The rate of decline in neutralizing antibody production in patients responding to replacement therapy was The first level of neutralizing antibodies produced in response to replacement therapy prior to administration of the dose is measured using the first or second In response to replacement therapy after the 3rd, 4th, or 5th dose of VIB4920 and a second level of neutralizing antibodies produced in response to the antibody or Alternatively, the level of neutralizing antibodies produced by the patient in response to the replacement therapy may be determined. The rate of decrease in the peaks produced in response to replacement therapy before the first dose of VIB4920 was Neutralizing antibody levels of VIB49 after the first, second, third, fourth, or fifth round of vaccination were Compared to peak neutralizing antibody levels produced in response to replacement therapy after 20 doses of The concentration may be determined by comparing the concentration of the

[0029] Alternatively, or in addition, inducing immune tolerance to replacement therapy in a patient may include administering a replacement The objective of the present invention may be to reduce the T cell response to the therapy. or if replacement therapy has been administered but there is still no T cell-mediated immune response to the replacement therapy, IB4920 may help prevent disease by blocking the formation of early T cell responses to replacement therapy. However, this can reduce the T cell response of patients with existing immune deficiencies. In patients with a T cell response to replacement therapy, VIB4920 reduces the existing T cell response to replacement therapy. By reducing the number of T cells in the blood, immune tolerance can be induced. At least 50%, at least 55%, at least 60%, at least 65%, at least at least 70, at least 75%, at least 80%, at least 85%, at least 90 %, at least 95%, or to undetectable levels. The rate of decline in T cell responses to refill therapy was significantly greater with the initiation of refill therapy prior to the first VIB4920 dose. The first level of T cell response to The second level of T cell response to replacement therapy after administration of a dose of VIB4920 was compared with It may be a comparison of, or may be determined by, the patient. The rate of decline in T cell response to replacement therapy in patients was significantly higher than that in patients with replacement therapy before the first dose of VIB4920. Peak T cell response levels to full therapy were measured after the first, second, third, fourth, or Peak T cell response levels to replacement therapy after the fifth dose of VIB4920 The T-cell concentration may be determined by comparing the T-cell concentration with the T-cell concentration. Decreased cell response is defined as a decrease in proliferation of CD4+ T cells stimulated by replacement therapy and / or The reduced T cell response may be characterized by reduced stimulation. The tumor may be characterized by a decreased CD4-dependent CD8+ T cell response to the tumor.

[0030] The replacement therapy to which immune tolerance is induced may be a peptide or protein replacement therapy. If the replacement therapy is a peptide or protein therapy, factor VIII or factor IX therapy and may be administered to treat patients suffering from hemophilia. If it is a peptide or protein therapy, it may be an enzyme replacement therapy (ERT). If the treatment is ERT, replacement therapy is agalsidase alfa or agalsidase beta It may be possible to substitute α-galactosidase A, Patients suffering from Bryonia can be treated with ERT. The replenishing agent may be iduronidase, which replaces α-L-iduronidase. It is possible to treat mucopolysaccharidosis (MPS) type 1 (depending on its severity, Hurler syndrome, Patients with Hurler-Scheie syndrome (also known as Scheie syndrome) When replacement therapy is ERT, the replacement therapy is alglucosidase inhibitor. It may be an enzyme that can replace α-glucosidase, If the replacement therapy is ERT, the replacement therapy may be idursulfase, thereby iduronate-2-sulfatase It is possible to replace the IL-11 gene and treat patients with MPS II. If the replacement therapy is ERT, the replacement therapy is imiglucerase or velaglucerase. alpha or taliglycerase alpha, which can enhance the production of β-glucocerebrose. It is possible to replace the sidase and treat patients with Gaucher disease. If the replacement therapy is ERT, the replacement therapy is Naglazyme arylsulfate. may be N-acetylgalactosamine-4-sulfatase B, thereby replacing N-acetylgalactosamine-4-sulfatase It is possible to convert the IL-11 receptor agonist to IL-11 receptor agonist and treat patients suffering from MPS VI. When the peptide or protein replacement therapy is a peptide or protein, it is effective in inducing immune tolerance. Can the production of neutralizing antibodies against this peptide or protein be reduced? and / or reducing T cell responses to the peptide or protein by the patient. can.

[0031] Furthermore, tolerance-inducing replacement therapies can be used with recombinant vectors encoding therapeutic peptides or proteins. The replacement therapy may be a viral vector containing the nucleic acid. When the viral vector comprises a nucleic acid encoding an adenovirus, the viral vector is vectors, adeno-associated virus vectors, retrovirus vectors, poxvirus , alphavirus, herpes simplex virus vectors, or therapeutic peptides or proteins Other viral vectors capable of delivering nucleic acids encoding proteins to the patient's cells. The viral vector may be, for example, pseudotyped. and / or to delete the wild-type gene thereof, and / or to administer a therapeutic peptide or protein. The vector may be modified to contain a nucleic acid encoding a protein.

[0032] The therapeutic peptide or protein encoded by the nucleic acid of the viral vector is a therapeutic peptide or protein for Factor III or Factor IX; or Agalsidase alpha, agalsidase beta, idursulfase, ialonidase alglucosidase, imiglucerase, velaglucerase alpha, tali ERTs such as glycerase alpha or Naglazyme arylsulfatase B It's fine.

[0033] Additionally, replacement therapy may involve the use of a viral vector containing a nucleic acid encoding a therapeutic peptide or protein. VIB4920 may reduce the immune response to viral vectors. and a therapeutic peptide or protein encoded by a viral vector. Induce immune tolerance by reducing immune responses to VIB4920 can inhibit the vector itself or cells infected with the viral vector. The present invention provides a method for reducing neutralizing antibody and / or T cell responses to any viral vector in a patient. In addition, or by administering the same to the infected subject, immune tolerance to the viral vector can be induced. Instead, VIB4920 is a therapeutic antibody encoded by nucleic acid in a viral vector. By reducing neutralizing antibody or T cell responses to the peptide or protein, It is possible to induce immune tolerance to replacement therapy consisting of a virus vector.

[0034] VIB4920 used in various methods has the amino acid sequence shown in FIG. VIB4920 may have the amino acid sequence as shown in FIG. or one or more amino acid residue modifications to the amino acid sequence shown in FIG. VIB4920 may have an amino acid sequence similar to that shown in FIG. When having an acid sequence modification, the modification may be to one of the linkers VIB4920 amplifies the Gly15 linker that separates the two CD40L-specific monomers. and a Gly10 linker separating the CD40L-specific monomer from the HSA sequence. Either or both of these linkers may be altered, and the amino acid sequence (G m X) n [wherein X is serine (S), alanine (A), glycine (G), leucine (L), isopropyl alcohol (A), isine (I) or valine (V), m and n are integer values, m is 1, 2, 3 or 4 and n is 1, 2, 3, 4, 5, 6, or 7. For example, One or both linkers may be Amino acids containing one of the following: GGGGGGGGGG or GGGGGGGGGGGGGG VIB4920 may be altered to have the amino acid sequence shown in FIG. If the amino acid sequence of the CD40L-specific monomer is related to that of the CD40L-specific monomer, the CD40L-specific monomer is fused to the CD40L-specific monomer. The two CD40L-specific HSAs may be derived from modified or altered amino acid sequences. The HSA fused to the target monomer is derivatized to the full-length mature HSA except for at least one amino acid substitution. Numbered for positions, 407, 415, 463, 500, 506, 508, 509, 511, 512, 515, 516, 521, 523, 524, 526, 535, At a position selected from the group consisting of 550, 557, 573, 574, and 580, The CD40L specific Tn3 monomer may be fused to HSA, At least one amino acid substitution is a substitution of lysine (K) at position 573 with glutamic acid ( E) substitutions to the amino acid sequence shown in FIG. In the case of a polypeptide having an alteration in the amino acid sequence, the alteration may adversely affect the efficacy of VIB4920 in vivo. Unless otherwise provided, the amino acid sequence of one or both of the CD40L-specific Tn3 monomers. For example, one or both of the CD40L-specific Tn3 monomers may be directed against The amino acid sequence may be modified to have the amino acid sequence shown in FIG. 19A. stomach.

[0035] The dose of VIB4920 administered in this method is about 500 mg to about 3000 mg. The dose may be about 750 mg to about 3000 mg, or about 1000 mg to about 3 000mg, or about 1500mg to about 3000mg, or about 500mg to about 2000mg , or about 750 mg to about 2000 mg, or about 1000 mg to about 2000 mg, or about 1 The dosage may be from about 1,000 mg to about 2,500 mg, or from about 1,000 mg to about 1,500 mg. Available in 500mg, 750mg, 900mg, 1000mg, 1250mg, and 1500mg , 1750mg, 2000mg, 2250mg, 2500mg, or 3000mg That's fine.

[0036] Doses of VIB4920 may be administered approximately every other week, or twice a month. Alternatively, the dose of VIB4920 may be administered approximately weekly or approximately once a month. The dose of VIB4920 may be administered every 7 days, every 10 days, every 14 days, every 15 days, or every 16 days. VIB4920 may be administered every 14 to 10 days, every 14 to 16 days, or every 30 days. may be administered by intravenous or subcutaneous injection.

[0037] The administered dose of VIB4920 is 1000 mg, 1500 mg, or about 1000 mg. If the dose is between about 1500 mg and about 1500 mg, the dose may be administered every other week or once a month. If the dose of VIB4920 is 3000 mg, VIB4 VIB4920 may be administered once a month at a dose of 500 mg. or 750 mg, the dose of VIB4920 may be administered once every other week; or Alternatively, it may be administered twice a month. Either of these doses may be administered intravenously. good.

[0038] The dose and dosing regimen of VIB4920 is suitable for treating any autoimmune / inflammatory disease or disorder. Any therapeutic effect achieved by administering VIB4920 to Decreased autoantibodies, decreased Vectra DA score, decreased plasma cell signature, CR Decreased P, decreased DAS28-CRP, decreased number of swollen joints, decreased number of tender joints, CDAI reduction in schizophrenia, improvement in patient global assessment, improvement in physician global assessment, achievement of ACR20, Achieving CR50 or ACR70 can be considered to be “sustained for a long time.” VIB4920 in the treatment of autoimmune / inflammatory diseases or disorders. The "long-lasting" effect of VIB4920 means that the therapeutic effect achieved by VIB4920 is After the last dose of 4920 has been administered (VIB4920 is no longer being administered), Nevertheless) at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks , at least 12 weeks, at least 16 weeks, at least 20 weeks, or at least 24 weeks A course of VIB4920 should be administered every 7 to 31 days (e.g., every 7 days). every 10 days, every 14 days, every 15 days, every 16 days, every 14 to 10 days, every 14 to 16 days, or Once every 30 days) for about 8 to 24 weeks (e.g., 8 weeks, 10 weeks, or 12 weeks) weeks, or 14 weeks, or 16 weeks, or 18 weeks, or 20 weeks, or 22 weeks, or 24 weeks, or 500mg to 3000mg (e.g. 2 months, 4 months, or 6 months) For example, 500mg, 750mg, 1000mg, 1250mg, 1500mg, 1750 mg, 2000mg, 2250mg, 2500mg, 2750mg, or 3000mg) The dose of VIB4920 may be administered.

[0039] The present disclosure encompasses the following embodiments. [Section 1] 1. A method for suppressing a B cell mediated immune response in a subject, comprising: Doses of 500mg to 3000mg of VIB4920 will be administered to subjects in need. To do, and suppressing the B cell mediated immune response. [Section 2] Item 1, wherein the dose is 1000 mg to 1500 mg of VIB4920. . [Section 3] 3. The method of claim 2, wherein the dose is 1000 mg of VIB4920. [Section 4] 3. The method of claim 2, wherein the dose is 1500 mg of VIB4920. [Section 5] The method according to any one of claims 1 to 4, wherein the dose is administered every 14 days or twice a month. 2. The method according to any one of claims 1 to 11. [Section 6] 6. The method according to any one of items 1 to 5, wherein the dose is administered intravenously. [Section 7] The suppressing of the B cell mediated immune response reduces antibody class switching. The method according to any one of Items 1 to 6, wherein [Section 8] The suppression of the B cell-mediated immune response is a reduction in circulating B cells. 7. The method according to any one of 1 to 6. [Section 9] The suppressing of the B cell mediated immune response is a decrease in plasma cell activity. Item 7. The method according to any one of Items 1 to 6. [Section 10] wherein the reduction in plasma cell activity is a reduction in a plasma cell signature. Item 10. The method according to any one of items 9. [Section 11] The reduction in the plasma cell signature is due to the expression of the genes IGHA1, IGJ, IGKC, IGKV Item 11. The method according to item 10, characterized in that the expression of TNFRSF17, TNFRSF17, and TNFRSF17 is decreased. Law. [Section 12] 1. A method for treating an autoimmune disease or disorder, comprising: Doses of 500 mg to 3000 mg of VIB4920 will be administered to patients in need. And, treating said autoimmune disease or disorder. [Section 13] Item 13. The method according to item 12, wherein the dose is 1000 mg to 1500 mg of VIB4920. Law. [Section 14] 14. The method of claim 13, wherein the dose is 1000 mg of VIB4920. [Section 15] 15. The method of claim 14, wherein the dose is 1500 mg of VIB4920. [Section 16] The dose is administered every 14 days or twice a month. 6. The method according to any one of claims 5 to 5. [Section 17] Item 17. The method according to any one of items 12 to 16, wherein the dose is administered intravenously. [Section 18] The method further comprises the steps of: treating the autoimmune disease or injury by reducing an inflammatory marker; Item 18. The method according to any one of Items 12 to 17, wherein the method is characterized by the above. [Section 19] The inflammatory markers include autoantibody levels, plasma cell (PC) signatures, circulating B cells, and antibody class switching. [Section 20] 20. The method of claim 18, wherein said treating is reducing a clinical symptom. [Section 21] Item 13. The method of item 12, wherein the autoimmune disease or disorder is rheumatoid arthritis. [Section 22] The treatment includes treating rheumatoid factor (RF) autoantibodies, Vectra DA Bio Marker score, plasma cell (PC) signature, serum reactive C protein (CRP), DAS28-CRP, swollen joint count, tender joint count, or Clinical Disease Activity Index (CDAI) Item 22. The method according to item 21, wherein one or more of the following is reduced: [Section 23] Item 22, wherein the treating is to reduce rheumatoid factor autoantibodies. Method of posting. [Section 24] The reduction in RF autoantibodies is at least 50% within 85 days of starting treatment. Item 24. The method according to item 23, wherein [Section 25] The method according to item 22, wherein the treating is to reduce DAS28-CRP. Law. [Section 26] The reduction in DAS28-CRP was defined as an adjusted mean difference of at least -1.2. Item 26. The method according to item 25, wherein [Section 27] The decrease in DAS28-CRP was detectable following administration of a single dose of VIB4920 Item 26. The method according to item 25, wherein [Section 28] The treating is characterized in that it reduces the Vectra DA biomarker score. 23. The method according to claim 22, [Section 29] The reduction in the Vectra DA biomarker score was defined as a small adjusted mean difference 29. The method according to claim 28, wherein the .lambda. [Section 30] The autoimmune disease or disorder is systemic sclerosis, myositis, antiphospholipid syndrome, autoimmune hepatitis, lupus nephritis, idiopathic thrombocytopenic purpura, vasculitis, cutaneous lupus, autoimmune Hemolytic anemia, Sjogren's disease, IgG4-related disease, or systemic lupus erythematosus Item 13. The method according to item 12, which is one of the methods. [Section 31] said treating is reducing clinical symptoms of said autoimmune disease or disorder; 31. The method according to item 30. [Section 32] 31. The method of claim 30, wherein said treating is reducing a PC signature. . [Section 33] The reduction in the PC signature was observed in the genes IGHA1, IGJ, IGKC, IGKV4- 33. The method according to claim 32, characterized in that the expression of TNFRSF17 is reduced. [Section 34] The treating reduces one or more biomarkers of the autoimmune disease or disorder. Item 31. The method according to item 30, wherein the method is [Section 35] Reduces indicators of rheumatoid arthritis (RA) disease activity in patients undergoing treatment for RA A method for making the administering to said patient VIB4920, The indicators of RA disease activity are DAS28-CRP, Clinical Disease Activity Index (CDAI ), patient global assessment, or physician global assessment; said VIB4920 being administered at a dose of about 500 mg to 3000 mg; and reducing said indicator of RA disease activity in said patient. [Section 36] Item 35. The VIB4920 is administered at a dose of about 1000 mg to 2000 mg. The method described. [Section 37] Item 36. The method according to item 36, wherein the VIB4920 is administered at a dose of about 1000 mg to 1500 mg. The method described. [Section 38] 38. The method of claim 37, wherein the VIB4920 is administered at a dose of about 1000 mg. [Section 39] 38. The method of claim 37, wherein the VIB4920 is administered at a dose of about 1500 mg. [Section 40] wherein the VIB4920 is administered every 14 days or twice a month; Item 40. The method according to any one of items 35 to 39. [Section 41] Item 41. The method according to any one of items 35 to 40, wherein the VIB4920 is administered intravenously. . [Section 42] The index is DAS28-CRP, and the reducing refers to an adjusted mean change of 42. The method according to any one of items 35 to 41, wherein the ratio is at least −1.2. [Section 43] The decrease is an adjusted mean change of at least -2.2. 43. The method according to item 42. [Section 44] The index is DAS28-CRP, and the reduction is observed after the first dose of VIB4920. Item 42. The method according to any one of Items 35 to 41, [Section 45] Reduces rheumatoid factor (RF) autoantibodies in patients undergoing treatment for rheumatoid arthritis 1. A method for detecting a cellular function of a blood vessel comprising: administering to the patient a dose of about 500 mg to 3000 mg of VIB4920; and reducing RF autoantibodies in said patient. [Section 46] Item 46. The method according to item 45, wherein the dose is about 1000 mg to about 2000 mg. [Section 47] Item 47. The method according to item 46, wherein the dose is about 1000 mg to about 1500 mg. [Section 48] 48. The method of claim 47, wherein the dose is about 1000 mg. [Section 49] 49. The method of claim 48, wherein the dose is about 1500 mg. [Section 50] Item 45-4, wherein the dose is administered every 14 days or twice a month. 10. The method of any one of claims 9 to 9. [Section 51] 51. The method of any one of items 45 to 50, wherein the dose is administered intravenously. [Section 52] Item 52. The method according to any one of items 45 to 51, wherein the RF autoantibody is reduced by at least 40%. The method described. [Section 53] Item 46 to 51, wherein the RF autoantibody is reduced by at least 50%. The method described. [Section 54] Item 46 to 53, wherein the RF autoantibody is reduced within 85 days after the start of treatment. 13. The method according to claim 1. [Section 55] Methods for reducing biomarker scores in patients undergoing treatment for rheumatoid arthritis - Patent Application 20100223333 There was, administering to the patient about 500 mg to 3000 mg of VIB4920, The biomarker scores include the Plasma Cell (PC) gene signature, Vectra - DA score or serum C-reactive protein level (CRP) And, and decreasing said biomarker score in said patient. [Section 56] Item 56. The method according to item 55, wherein the dose is about 1000 mg to about 2000 mg. [Section 57] Item 57. The method according to item 56, wherein the dose is about 1000 mg to about 1500 mg. [Section 58] 58. The method of claim 57, wherein the dose is about 1000 mg. [Section 59] 59. The method of claim 58, wherein the dose is about 1500 mg. [Section 60] The dose is administered every 14 days or twice a month. 10. The method of any one of claims 9 to 9. [Section 61] Item 61. The method of any one of items 55 to 60, wherein the dose is administered intravenously. [Section 62] the biomarker score is Vectra DA and the decreasing is The adjusted mean difference is at least -10.3 for any one of items 55 to 61. The method described. [Section 63] Item 62. The method according to any one of Items 57 to 61, wherein the biomarker score is serum CRP. How to. [Section 64] To reduce the plasma cell (PC) gene signature score in patients who need it The method of claim 1, administering VIB4920 to a patient in need thereof; The patient may have systemic lupus erythematosus, rheumatoid arthritis, myositis, antiphospholipid syndrome, , autoimmune hepatitis, or Sjogren's disease, said VIB4920 being administered at a dose of about 500 mg to 3000 mg; and decreasing said patient's PC gene signature score. [Section 65] Item 65. The method according to item 64, wherein the dose is about 1000 mg to about 2000 mg. [Section 66] Item 66. The method according to item 65, wherein the dose is about 1000 mg to about 1500 mg. [Section 67] 67. The method of claim 66, wherein the dose is about 1000 mg. [Section 68] 68. The method of claim 67, wherein the dose is about 1500 mg. [Section 69] 64 to 6, wherein the dose is administered every 14 days or twice a month. 9. The method according to any one of claims 8 to 9. [Section 70] 70. The method of any one of claims 64 to 69, wherein the dose is administered intravenously. [Section 71] 1. A method for reducing autoantibodies in a patient undergoing treatment for an autoimmune disorder, comprising: administering VIB4920 to a patient in need thereof; the patient is being treated for an autoimmune disease characterized by the presence of autoantibodies; said VIB4920 being administered at a dose of about 500 mg to 3000 mg; and reducing said autoantibodies in said patient. [Section 72] Item 72. The method according to item 71, wherein the dose is about 1000 mg to about 2000 mg. [Section 73] Item 73. The method according to item 72, wherein the dose is about 1000 mg to about 1500 mg. [Section 74] 74. The method of claim 73, wherein the dose is about 1000 mg. [Section 75] 74. The method of claim 73, wherein the dose is about 1500 mg. [Section 76] The dose is administered every 14 days or twice a month. 6. The method according to any one of claims 5 to 5. [Section 77] 77. The method of any one of claims 71 to 76, wherein the dose is administered intravenously. [Section 78] The autoimmune disease is systemic lupus erythematosus, rheumatoid arthritis, myositis, antiphospholipid antibody 77. The disease described in any one of items 71 to 77 is hepatitis, autoimmune hepatitis, or Sjogren's disease. Method of posting. [Section 79] 1. A method for reducing inflammation in a patient, comprising: administering VIB4920 to a patient in need thereof; The patient is being treated for an inflammatory disease or disorder, or is undergoing organ or tissue transplantation. The patient is receiving preventative treatment for the inflammation that is predicted in response. said VIB4920 being administered at a dose of about 1000 mg to 3000 mg; and reducing inflammation in said patient. [Section 80] Item 80. The method according to item 79, wherein the dose is about 1000 mg to about 2000 mg. [Section 81] Item 81. The method according to item 80, wherein the dose is about 1000 mg to about 1500 mg. [Section 82] 80. The method of claim 79, wherein the dose is about 1000 mg. [Section 83] 80. The method of claim 79, wherein the dose is about 1500 mg. [Section 84] 80. The method of claim 79, wherein the dose is about 3000 mg. [Section 85] The dose is administered every 14 days or twice a month. 5. The method according to any one of claims 4 to 4. [Section 86] 86. The method of any one of clauses 79 to 85, wherein the dose is administered intravenously. [Section 87] 85. The method of claim 84, wherein said dose is administered once per month. [Section 88] the patient is being treated with the combination to prevent rejection of an organ or tissue transplant; 88. The method according to any one of items 79 to 87, wherein the treatment is prophylactic or preventive. [Section 89] The inflammatory disease or disorder is an inflammatory myopathy, or lupus nephritis, cutaneous lupus, RA, SLE, ITP, myositis, Sjogren's syndrome, vasculitis, systemic sclerosis, autoimmune hemolytic Anemia, myasthenia gravis, or focal segmental glomerulosclerosis, according to any one of items 79 to 87. The method described. [Section 90] The treating is achieving ACR20, ACR50, or ACR70. 22. The method according to claim 21. [Section 91] Item 12. The method according to any one of items 1 to 11, wherein the suppression of the B cell-mediated immune response lasts for a long period of time. Method of posting. [Section 92] The method according to any one of claims 12 to 34 or 90, wherein the treatment of the autoimmune disease or injury lasts for a long time. The method according to any one of claims 1 to 5. [Section 93] Item 35 to 44, wherein the reduction in the index of RA disease activity in the patient is sustained over time. 2. The method according to any one of claims 1 to 11. [Section 94] 55. Any one of claims 45 to 54, wherein the reduction in RF autoantibodies in the patient is sustained over time. The method according to [Section 95] Any of items 55 to 63, wherein the decrease in the biomarker in the patient is sustained over time. 3. The method according to claim 1 . [Section 96] Item 64. The reduction in the patient's PC gene signature score is sustained over time. A method according to any one of claims 1 to 70. [Section 97] 79. Any one of claims 71 to 78, wherein the reduction in the autoantibodies in the patient is sustained over time. The method according to [Section 98] 89. The method according to any one of claims 79 to 89, wherein the reduction in inflammation in the patient is sustained over time. method. [Section 99] 1. A method of inducing immune tolerance to replacement therapy in a patient, comprising: administering VIB4920 to a patient in need of replacement therapy, said VIB4920 being administered at a dose of about 1000 mg to 3000 mg; and reducing immune tolerance to said replacement therapy in said patient. [Section 100] Item 99. The method according to item 99, wherein the VIB4920 is administered at a dose of about 1500 mg to 3000 mg. The method described. [Section 101] Item 100, wherein the VIB4920 is administered at a dose of about 2500 mg to 3000 mg. The method according to [Section 102] 102. The method of claim 101, wherein the VIB4920 is administered at a dose of about 3000 mg. [Section 103] Item 99-102, wherein the dose is administered approximately once every 2-4 weeks. method. [Section 104] 104. The method of claim 103, wherein said dose is administered about once every four weeks. [Section 105] 103. The method of claim 102, wherein said dose is administered once per month. [Section 106] Item 99, 102 or 105, wherein the replacement therapy is a protein or peptide. 3. The method according to claim 1 . [Section 107] Inducing immune tolerance to the protein or peptide by the patient. The method of claim 106, comprising reducing the production of neutralizing antibodies. [Section 108] 108. The method according to claim 107, wherein the protein is factor VIII and the patient is a hemophilia patient. Method of posting. [Section 109] 108. The method of claim 107, wherein the protein is factor IX and the patient is a hemophilia patient. method. [Section 110] Inducing immune tolerance reduces neutralizing anti-Factor VIII antibodies in the patient. Item 109. The method according to Item 108, wherein the method comprises: [Section 111] 107. The method of claim 106, wherein the protein or peptide is an enzyme. [Section 112] the enzyme is agalsidase alpha or agalsidase beta, and the patient is The method of claim 111, wherein the patient is a Brey disease patient. [Section 113] The enzyme is idursulfase, and the patient is a mucopolysaccharidosis II patient or a hunter's disease patient. The method according to claim 111, wherein the patient is a patient with rheumatoid arthritis syndrome. [Section 114] Item 111. The enzyme is arylonidase, and the patient is a mucopolysaccharidosis type I patient. The method described. [Section 115] Item 1. The enzyme is alglucosidase alpha, and the patient is a Pompe disease patient. 11. The method according to claim 11. [Section 116] The replacement therapy may comprise a viral vector containing a nucleic acid encoding a therapeutic peptide or protein. Item 99. The method of any one of items 99, 102, or 105, wherein the target is a catalyzer. [Section 117] Inducing immune tolerance to the replacement therapy comprises administering to the patient the viral vector. reducing the immune response to said therapeutic protein; 117. The method of claim 116, comprising: [Section 118] 118. The method according to claim 117, wherein the viral vector is an adeno-associated virus (AAV). Law. [Section 119] Inducing immune tolerance includes reducing an immune response to the AAV. 119. The method according to claim 118. [Section 120] The reducing the immune response to the AAV includes increasing the T cell response to the AAV. Item 119. The method of claim 119, wherein the objective of the method is to reduce the response to the AAV or to reduce the antibody against the AAV. The method described. [Section 121] The reducing the T cell response includes reducing the T cell response to an AAV capsid protein. Item 121. The method of item 120, wherein the method is to reduce [Section 122] Inducing immune tolerance to the replacement therapy comprises inducing immune tolerance to the therapeutic protein. 118. The method of claim 117, comprising reducing neutralizing antibodies. Those of ordinary skill in the art will be able to readily identify, using no more than routine experimentation, the specific embodiments described herein. Numerous equivalents may be recognized or identified. Such equivalents are set forth in the following claims: It is intended that all such modifications and variations be encompassed by the scope of the claims.

[0040] All publications, patents and patent applications mentioned herein are hereby incorporated by reference in their entirety. Each and every patent or patent application is specifically and individually indicated to be incorporated by reference herein. and are incorporated by reference herein to the same extent as if fully set forth herein. EXAMPLES

[0041] Example 1 - Isolation and optimization of the CD40L-specific Tn3 protein Tn3 is a small protein scaffold approximately 90 amino acids long that is randomized and specifically Loops structurally similar to the complementarity determining regions of antibodies that can be used to select for distinct binding characteristics. It has an immunoglobulin-like fold, including ( 24 ).

[0042] As described in detail in WO 2013 / 055745 In 2003, human CD40L-specific Tn3 clones were isolated (see also 24, 27, 50 Briefly, the selection of human CD40L-specific Tn3 was performed using recombinant human CD40L protein. The study included five rounds of panning, alternating between the selection of a human CD40L-expressing CHO cell line and a human CD40L-expressing protein. The results showed that mouse CD40L was produced using only recombinant mouse CD40L protein. Select specific Tn3 proteins. Express Tn3 genes from selected outputs. The individual His-tagged variants were pooled and isolated using an anti-His antibody. By capturing onto coated Maxisorp plates (2 μg / ml in PBS) CD40L binding was assessed using biotinylated MegaCD40L (Enzo Bio Sciences, 0.5 μg / mL) was added and incubated for 1.5 hours. After washing once with S / Tween, SA-HRP (1:1000 dilution) was used for capture. The interaction between the captured Tn3 variants and CD40L was monitored. After 20 min, Plates were washed twice with PBS / Tween and developed with TMB substrate and 2.5M H3 The enzyme was stopped by PO4. The absorbance was measured at 450 nm. Select improved candidates from a phage-displayed library of mutants By using this method, we performed affinity maturation of the CD40L-specific Tn3 protein (24). The method produced an improved variant of human CD40L-specific 309 (Figure 21A). Lone 342 (Figure 19), and an improved variant of the mouse CD40L-specific M13 One clone, M31, was generated as a result. For example, clones 304, 311, 320, 310, 321 and 322 were also prepared. See WO 2013 / 055745, which is incorporated herein by reference. I want to be illuminated.

[0043] This set of human CD40L-specific Tn3 clones was analyzed by immunohistochemistry, in which CD40L binds to its receptor (CD 40) were characterized for their ability to biochemically inhibit binding to this set of All seven inhibited the binding of CD40L to CD40 with IC50 values ​​below 1 μM. The two most potent CD40L-CD40 inhibitors in the biochemical CD40L-CD40 inhibition assay Inhibitors were used to block CD40L-mediated signaling in cell-based reporter assays. The toxicity of human CD40 and NF-kB-luciferase reporter genes was further evaluated. HEK-293 cells expressing the CD40L gene were stimulated with recombinant human CD40L protein. CD40L-specific Tn3 proteins 309 and 311 are involved in CD40L-induced NF- It inhibited the expression of the kB reporter gene in a dose-dependent manner at micromolar concentrations (Figure 1B). highlighting the ability of these proteins to functionally inhibit CD40 / CD40L signaling. Ta.

[0044] Simultaneous binding to multiple targets, as occurs with bivalent antibodies, is greatly enhanced. This may result in a different avidity for the efficacy of the CD40L-specific Tn3 protein. To investigate the effect of valency, the same Tn3 module (309-309, e.g., Figure 2 1B) are linked via a flexible Gly4Ser containing spacer. In response to stimulation with CD40, primary human B cells expressed tandem bivalent fusion proteins. In response to this, expression of the activation marker CD86 is increased. Preincubation with monovalent CD40L-specific Tn3 309 resulted in the expression of human CD19+ B cells. The upregulation of CD86 on the cells was inhibited in a dose-dependent manner (Figure 1C). In primary cell assays using bivalent constructs, nearly 1000% more Tn3 was detected in primary cells compared to monovalent Tn3. A 2-fold increase in potency was observed (Figure 1C). In addition, clone 3, resulting in clone 342, Affinity maturation (by random mutagenesis in the variable CDR-like loop regions) of 09 This resulted in a marked increase in binding affinity to CD40L (309:190 nM, 342:1. 4 nM), which showed approximately 300-fold increased potency in blocking the CD40-CD40L interaction The affinity matured clone 342 was also shown to bind to Fas, TNFα, For binding to a panel of related TNF family members including TNFβ and OX40L By screening, it was found that CD40L selectively binds to the antibody. See FIG. 1E. sea ​​bream.

[0045] Finally, naked Tn3 molecules, like other alternative scaffolding technologies, are difficult to engineer due to their small size. Therefore, it is predicted to show very rapid clearance from the blood circulation when administered systemically. To improve the pharmacokinetic properties of this protein, The protein was fused to serum albumin (28, 29). The Tn3 protein, M13-M13, was expressed in mice in <30 minutes when delivered systemically. Mouse serum albumin (MSA) was added to the M13-M13 Tn3 protein. Fusion to the protein increased serum half-life by 65-fold and reduced clearance by 345-fold. The results were as follows (Table 1).

[0046] [Table 1]

[0047] Five- to seven-week-old CD-1 mice were injected with bivalent CD40L-specific A single injection of Tn3 molecule was administered (n=12 / group, 10 mg / kg, iv). Blood was collected from n=3 mice / group at various time points and Tn3 antigen was assayed by ELISA. Protein concentrations in the blood were measured.

[0048] Based on these observations, we developed a bivalent human CD40L-specific Tn3 molecule, VIB49 20 is a tandem 342 CD40L-specific Tn3 protein clone designed to optimize efficacy. It is composed of 1, 2, and fused to human serum albumin (HSA) to improve its half-life (Figure 1). F, Figure 18).

[0049] To better understand the molecular nature of the interaction between CD40L and VIB4920 To investigate the mechanism of action of CD40L-specific Tn3(342) and soluble CD40L, The protein was expressed, purified and cocrystallized, and the structure was measured at 2.8 Å resolution. The molecular structure of trimeric soluble CD40L complexed with Tn3 is shown in Figure 2A. The interface with 10 hydrogen bonds formed between the molecules (within a distance of 3.5 Å, Figure 2B) It is composed primarily of amino acids from the second modified loop of Tn3, including eight Initial molecular characterization has demonstrated that 342Tn3 mediates the interaction between CD40L and CD40. To visualize the details of its action, The structure of the CD40 / CD40L complex was superimposed with that of 342 / CD40L (Fig. 1 G). By superposition, 342 and CD40 share a common binding site on CD40L. Therefore, VIB4920 competes with CD40 and binds to CD4 0 and CD40L.

[0050] Example 2 - VIB4920 blocks human B cell activation and differentiation CD40 signaling has been extensively characterized and involves several different pathways and NF-kB. It is involved in the activation of transcription factors including the IL-1 receptor activator (30), which mediates B cell activation, proliferation, and differentiation. Therefore, human CD40 and NF-kB luciferase can promote cell proliferation (31). CD40L-mediated activation of NF-kB using cell lines expressing a reporter gene The ability of VIB4920 to inhibit recombinant human CD40L or C Stimulation with D40L expressing cells induces activation of NF-kB. As evidenced by the dose-dependent inhibition of activation of B (IC50: 0.899 nM, Figure 3A), VIB4920 potently blocked CD40 signaling using this cell line. I was able to do it.

[0051] Resting B cells constitutively express low levels of the costimulatory molecule CD86, which is rapidly upregulated after activation, including activation by CD40 (32). BMCs were stimulated with recombinant human CD40L and analyzed by flow cytometry 16 hours later. The expression of CD86 on primary human B cells was assessed. VIB4920 inhibited the expression of CD86 on primary human B cells. completely blocked CD40L-mediated upregulation of β-lactam IgG1 (Fig. 3B).

[0052] Example 3 - VIB4920 does not induce platelet aggregation in vitro Anti-CD40L-directed mAbs are primarily targeted to CD40L on the cell surface of platelets due to safety concerns. Clinical trials have failed due to thromboembolic complications associated with cross-linking of Fc domain 40L. To confirm that VIB4920 lacking phosphokinase does not induce platelet aggregation, The authors evaluated its effect on washed human platelets in vitro. Anti-CD40L mAb (human IgG1) inhibited the proliferation of sCD40L in plasma. The reaction was rapid and the mAb-sCD4 The OL immune complex induced 80% of platelets to aggregate within 8 minutes. Platelets were preincubated with an antibody (mAb IV.3) that blocks FcγRIIa. This prevents mAb-immune complex-mediated agglutination, which is This is consistent with an essential role for Fc receptors in the regulation of IL-1 expression (Figure 3F). At some concentrations, VIB4920 showed no tendency to induce platelet aggregation in this assay. These data are not consistent with the absence of an Fc region in the Tn3 construct. , Platelet aggregation and risk of thromboembolic events observed with therapeutic anti-CD40L antibodies in clinical practice This suggests that the effect of the stimulant was able to be reduced by up to 300%. From a chronic (7-month) study in non-human primates administered VIB4920 at 1 mg / kg Data from (not shown) did not reveal any adverse findings on platelet function, e.g., D-dimer -test (to monitor for blood clots), PFA100 test (to evaluate platelet function), or No adverse findings were identified in the TAT complex (thrombin-antithrombin complex) test. It was not done.

[0053] Example 4 - CD40L-specific Tn3 protein regulates immune responses in vivo The central role of CD40L in promoting T-dependent immune responses has been well characterized. (9, 35). Therefore, we used a T-dependent immunity model to investigate the humoral immune response in vivo. The ability of the Tn3-MSA fusion protein to block the response to human CD40L and mouse These studies did not include CD40L-specific markers because there is insufficient sequence homology between CD40L and CD40L. A mouse alternative Tn3, M31, was used.

[0054] To determine whether the Tn3-MSA fusion protein can block immune responses in vivo, To verify this, mice were inoculated with sheep red blood cells (SRBCs) and then incubated for 9–13 days after inoculation. Eyes were treated daily with anti-CD40L Tn3 protein. Spleen The immune response of treated animals was assessed by quantifying germinal center B cells in the lymph nodes and lymph nodes. The mice were assessed on day 4. As expected, immunization with SRBCs from control-treated mice did not result in the development of germinal centers. This resulted in a significant expansion of the expression frequency of CD40L-specific Tn3-MSA fusion proteins (Figure 4A). A dose-dependent decrease in the frequency of germinal center B cells was observed in mice treated with ribozyme. The CD40L-specific Tn3-MSA fusion protein was induced at 30 mg / kg. At a dose of 1 g, germinal center B cells in the spleen and lymph nodes were nearly equal to those in control non-immunized mice. The drug administration induced a complete inhibition of germinal center formation, as assessed by the absence of This ensures that other subpopulations of cells, including specific T cell populations, are not perturbed and that the observed effects are This confirmed that this was not secondary to T cell depletion (data not shown). In addition, anti-SRBC IgG levels reflect the level of embryonic B cell responses, and SRB C-specific Ig titers were significantly reduced with higher doses of anti-CD40L Tn3 (Fig. 4B ).

[0055] Example 5 - VIB4920 is well tolerated in healthy volunteers In a Phase 1a study conducted in healthy adults aged 18 to 49 years, The safety profile of 4920 was evaluated in humans. Subjects were randomly assigned to receive up to 3,000 mg of VIB4. Patients were enrolled in seven single ascending dose cohorts of VIB4920 or placebo. Treatment-emergent adverse events (TEAEs) and The incidence of treatment-emergent serious adverse events (TESAEs) was used to assess the primary endpoint and safety. The efficacy and tolerability of TEAEs were measured. Across all dose cohorts, TEAEs were generally clinically significant. The severity of symptoms was mild, and the most common events included nasopharyngitis (common cold) and headache ( Table 2).

[0056] [Table 2]

[0057] The most common adverse events occurring in at least two subjects in a Phase 1a study in healthy volunteers were Commonly seen TEAE

[0058] Importantly, the overall proportion of subjects with one or more drug-related TEAEs was The overall incidence was comparable between the VIB4920 (40.9%) and placebo (33.3%) groups. In addition, there were no infusion reactions, serious infections, or deaths, and only one TE in the placebo group. Only one SAE, tibial fracture, was reported. Of note, in this Phase 1a study, No clinically significant coagulation or platelet function abnormalities were observed following treatment with IB4920 .

[0059] Example 6 - VIB4920 exhibits a favorable PK / PD profile in healthy volunteers Indicates the In addition to evaluating the safety profile of VIB4920, Pharmacokinetic (PK) and pharmacodynamic (PD) endpoints were also evaluated. The PK profile of VIB4920 following intravenous administration showed increased dose-related changes with increasing exposure. The mean terminal half-life of the molecule was 8 days at the highest dose. The half-life was up to 10.1 + / - 1.87 days.

[0060] CD40L is a transmembrane protein that is cleaved by both activated T cells and platelets. Soluble CD40L (sCD40L) is a 18KDa trimer. It is found at low levels in healthy donors and is elevated in the circulation of patients with autoimmune diseases. (36, 37) Measurement of sCD40L levels after VIB4920 administration Potentially, sCD40L bound to 4920 may be retained and accumulated in the blood circulation. As expected, plasma total sCD40L was significantly increased after VIB4920 administration. There was a dose-dependent increase in total sCD in plasma (Figure 6B), suggesting target binding. The time to reach the maximum 40L increased with increasing dose from 3 mg to 3000 mg. The duration of target binding increased from 11.5 to 84 days at the highest dose, and target binding was maintained for a longer period. It was shown that it was held.

[0061] Example 7 - Decreased ADA in Healthy Subjects Receiving Higher Doses of VIB4920 Observed Biologics are highly specific / selective in nature, but they do induce an immune response. Anti-drug antibodies (ADA) are a measure of the immunogenicity of therapeutic agents. In healthy volunteers, the majority of patients receiving low doses of VIB4920 ADA was detected (Figure 7). More specifically, in the dose range of 3 to 100 mg, Eighteen of the 20 subjects had detectable ADA, with 10 of those individuals being high The ADA titers were greater than the median titer of 480. In contrast, the incidence of ADA was significantly decreased at higher dose levels of VIB4920 (Figure 7), with Only 1 of 8 subjects developed detectable antidrug titers in the high-dose VIB study. The reduction in ADA expression observed with 4920 supports an immunomodulatory function of the molecule. In addition, the low rates and titers of ADAs may be due to the presence of better tolerated and more effective therapeutic agents. It is possible to convert it to.

[0062] Example 8 - VIB4920 inhibits T cell-dependent antibody responses in healthy volunteers to be harmed VIB4920 will be further evaluated for its ability to affect humoral immune responses in healthy subjects. This evaluation was performed by immunization with keyhole limpet hemocyanin (KLH). The effect of VIB4920 on T cell-dependent antibody responses (TDAR) induced by Healthy subjects in all treatment groups received two subcutaneous immunizations with KLH. (1) 14 days before administration of either VIB4920 or placebo and (2) 15 days after administration (Figure 5B). Both IgM and IgG antibodies were produced against KLH. were monitored for up to 113 days.

[0063] TDAR followed the expected trend in placebo-treated subjects, i.e., at 22 days (second Anti-KLH IgG titers increased dramatically within 1 week after immunization, reaching a peak level of IgG on day 29. was observed, followed by a decrease in KLH-specific IgG antibodies until the end of the monitoring period. Furthermore, consistent with previous reports, the 2 The anti-KLH response in the second challenge was predominantly IgG, and KLH-specific I was detected after rechallenge. The increase in gM was much more modest overall (Fig. 8B) (38-41).

[0064] As expected, healthy volunteers treated with low doses of VIB4920 showed significantly lower mortality rates than those treated with placebo. In contrast, patients treated with higher doses of VIB4920 had anti-KLH titers close to those of controls. Healthy volunteers started at a dose of 300 mg (p=0.035) and increased to 1,000 mg (p=0.002) and escalated doses of anti-KLH to 3,000 mg (p<0.001). There was a statistically significant decrease in the secondary response to KLH at day 43 in IgG. Of note, the 1000 mg and 3000 mg cohorts In the 43-day study, IgG against KLH was increased by 78% and 86%, respectively, compared with placebo. In the highest dose group, 7 of 8 subjects had decreased anti-KLH-IgG titers. Titers were undetectable at day 43, demonstrating near-complete suppression of VIB4920-induced humoral immune responses It was suggested that...

[0065] Example 9 - Immunosuppression of VIB4920 is mediated by inhibition of B cell proliferation and plasma cell responses Will be The mechanism by which VIB4920 suppresses secondary immune responses was determined by collecting peripheral blood from subjects before and after immunization. and characterizing circulating lymphocyte subsets by flow cytometry This became clearer when the secondary Immunization induced B cell proliferation, which was confirmed on the 22nd visit, i.e., rechallenge. We detected an increase in the frequency of Ki67+CD19+ B cells in the blood 7 days after the experiment. This was demonstrated by (Figure 9A).

[0066] In subjects who received a high dose of VIB4920 before rechallenge, compared with the placebo group The incidence of baseline B-cell proliferation was reduced in 11 patients with schizophrenia, suggesting that the proposed molecular mechanism Furthermore, the cohort that received the high dose of VIB4920 showed increased B cell proliferation after immunization. The cell proliferation response was significantly impaired, which was explained by the lack of an increase in Ki67+ B cells. See the 3000 mg cohort one week post-challenge (Figure 9A). Further phenotypic testing revealed that the greatest effect of VIB4920 on B cell proliferation was observed in Ig It was revealed that D-CD27+ isotype-switched memory populations These data suggest that VIB4920 significantly increased the IL-1 expression in response to a second challenge (Figure 9B). This is consistent with the TDAR results showing an inhibitory effect on IgG production.

[0067] Subjects treated with placebo or VIB4920 before and after secondary immunization with KLH We also monitored changes in gene expression in the peripheral blood of patients with glaucoma. It is an accurate and robust signature that can detect even the slightest changes in We used the plasma cell (PC) gene signature (42) to identify specific Consistent with the TDAR results, immunization resulted in a significant increase in platelet counts 1 week after rechallenge. A dramatic increase in the PC gene signature score in whole blood of Sebo-treated subjects was induced, The patients returned to baseline within 2 weeks (Figure 9C). 0 mg), the PC gene signature score in peripheral blood was This was significantly reduced compared to previous placebo-treated subjects (Figure 9C). In volunteers who received VIB4920, PC gene signature scores increased after reimmunization These data shed light on the mechanism of action of VIB4920 and its and demonstrates its potent ability to inhibit the PC response.

[0068] Example 10 - Repeated Doses of VIB4920 are Safe and Well Tolerated in RA Patients VIB4920 had an acceptable safety profile and was shown to be effective in healthy volunteers Mechanism of action, multiple ascending doses, and proof of concept demonstrated, making it a promising treatment for moderate to severe A Phase 1b clinical trial was conducted in adult patients with active RA. mg, n=8; 500mg, n=10; 1000mg, n=12; or 1500mg , n=12), or placebo (n=15) were administered intravenously (iv) every other week for 12 weeks. ) infusion (Figure 10). Patients were then observed for an additional 12 weeks after treatment. The primary endpoints measured at week 1 were safety, tolerability, PK parameters, ADA, and disease progression. Changes in disease activity (DAS28-CRP), RF autoantibodies, and serum C-reactive protein The study included additional biomarkers such as CRP and Vectra-DA scores. Fifty-three patients completed 12 weeks of treatment, with 2 patients (VIB4920 75 mg group) One patient in the VIB4920 1500 mg group and one patient in the VIB4920 1500 mg group discontinued treatment due to adverse events. One patient in the placebo group withdrew informed consent and was not assigned to VIB4920 75m One patient in group g was lost to follow-up. The biographical and clinical characteristics are shown in Figure 11.

[0069] Overall, VIB4920 demonstrated a significant improvement in T outcomes observed between placebo and the four active dose groups. It was generally safe and well tolerated due to its balanced distribution in EAE. The most common TEAEs were diarrhea, hyperhidrosis, upper respiratory tract infection, and urinary tract infection, respectively. occurred in 3 patients (7.1%). See Figure 12. Thrombotic adverse events, or No clinically significant coagulation abnormalities were observed. One adverse event (pronounced "encephalitis") occurred in the 1500 mg dose group after six doses of No etiological agent was identified, and similar symptoms were reported several months after discontinuing VIB4920. Similar symptoms recurred and the patient was subsequently diagnosed with metastatic melanoma of the brain.

[0070] Example 11 - VIB4920 exhibits a linear PK profile and reduces ADA in dose-dependent manner in RA patients Consistently reduced Similar to healthy Ph1a study volunteers who received low doses of VIB4920, ADAs were observed in RA patients receiving VIB4920. Three of eight RA patients (37.5%) received 0 mg of VIB4920, and three of eight RA patients (37.5%) received 500 mg of VIB4920. Three of the 10 RA patients (30%) who received 75 mg developed ADA (Figure 13B). In the VIB4920 dose group, two of eight subjects had detectable ADA during the treatment phase. All three subjects in the 500 mg VIB4920 treatment group developed detectable In the 1000 mg dose group, ADA was not detected during the treatment period (Figure 13C). No patients had any ADA after the treatment phase, and one subject had detectable ADA after the treatment phase. No ADA was detected in the 100-mL IgG4-treated control (Figure 13B), and VIB4920 effectively suppressed the ADA response at higher doses. It was suggested that this effectively suppresses

[0071] Example 12 - VIB4920 reduces disease activity in RA patients To determine whether VIB4920 reduces disease activity in RA patients in a Phase 1b clinical trial To confirm this, the DAS-28 / CRP score was measured. RA, taking into account swollen joint counts, tender joint counts, CRP levels, and patient global assessment. VIB4920 is a composite clinical disease activity score used in the treatment of chronic myocardial infarction and chronic myocardial infarction. Higher doses significantly reduced disease activity in RA patients as quantified by 8-CRP score (Figures 14A and 22A). Baseline DAS28-CRP at 12 weeks after the start of treatment The adjusted mean change (SE) from line was -2.3 in the VIB4920 1500 mg group. (0.3), VIB4920 1000 mg group -2.2 (0.3), VIB4920 -1.2 (0.3) in the 500 mg group and 0.1 (0.4) in the VIB4920 75 mg group. and −1.0 (0.3) in the placebo group (FIG. 14A). The reduction in DAS28-CRP scores in RA patients treated with VIB4920 was The effect was maintained for at least another 12 weeks after subsequent dosing (Figure 22A, specifically visit day 113) (See Days 141 and 169.) VIB4 for DAS28-CRP The effect of 920 was rapid, with reductions in scores evident within just 15 days after a single dose of the drug. It became.

[0072] Additionally, the reduction in disease activity at the two highest doses of VIB4920 was greater than that seen with placebo. The results were both clinically and statistically significant, with the adjusted mean difference (SE) at 12 weeks: was -1.4 (0.4) in the VIB4920 1500 mg group and -1.4 (0.4) in the VIB4920 1000 mg group. and -1.2 (0.4) in the 1 mg group, with p values ​​of 0.002 and 0.006, respectively. Using a linear dose-response model, a statistically significant dose-response effect on DAS28CRP was observed. A response was observed (p<0.001), primarily due to the 1000 mg and 1500 mg treatment groups. The 500 mg and 75 mg doses produced significant results, whereas the 500 mg and 75 mg doses produced little effect compared to placebo. Little or no effect was observed (Figure 15A). 75% of patients in the 1,000 mg dose group and 50% of patients in the 1,000 mg dose group had a score of 3.2 or less at week 12 The primary endpoint was to achieve a DAS28-CRP score of 0.01 and low disease activity or clinical The patient responded as normal, and showed a response. See FIG.

[0073] Example 13 - VIB4920 reduces immunological and inflammatory biomarkers in RA patients do Vectra DA blood test for immunological and inflammatory biomarkers The effect of VIB4920 was measured. The Vectra DA test is a 1-step analysis of disease activity. Two biomarkers (cell adhesion factor, growth factor, cytokine, matrix metalloproteinase) RA disease activity was measured using a 10-item scale (protease, skeletal proteins, hormones and acute phase proteins) These were combined into a single score to assess the major mechanisms and pathways that cause It is a commercially available and validated test that uses 1500mg and 1000mg of The 1.5-g dose of VIB4920 was administered every other week for a 12-week period (Figure 14 E) and a 12-week observation period when VIB4920 was no longer administered (Figure 22E). Both significantly reduced the Vectra DA multi-biomarker score. The adjusted mean difference (Week 12) for VIB4920 in mg doses compared with placebo was -14.4 (-21.5, -7.2), p=0.001, and the 1000 mg dose The adjusted mean difference (at week 12) for VIB4920 compared with placebo was -10.3 (- 17.4, -3.3), p = 0.018 (Figure 14E).

[0074] These efficacy results were consistent with other endpoints evaluated in the study, including the Clinical Disease Activity Index (CDA I) Measures included tender and swollen joint counts, patient and physician global assessments, and serum CRP levels. The results were very consistent across the board (including the 1000 mg and 1500 mg doses). This study confirmed that the dose was effectively effective (Figures 14B to 14D and Figure 1 7A-17C, 22B-22D and 23A-23C).

[0075] Example 14 - VIB4920 Significantly Reduces Rheumatoid Factor Autoantibodies in RA Subjects do Rheumatoid factor autoantibodies (RF) are autoantibodies produced against the Fc portion of IgG. They are increased in RA and are associated with poor prognosis. Taking into account the mechanism of action of 20, we evaluated their impact on autoantibody titers in RA subjects. Notably, VIB4920 increased the 500,000 mg / kg / day increase in serotonin levels during the 12-week, biweekly treatment period. It significantly reduced RF titers at the 1000 and 1500 mg dose levels (Figure 14F Even more surprising was the reduced RF titers at the 1000 mg and 1500 mg dose levels. was maintained throughout the 12-week observation period after the last dose of VIB4920 The reduction in RF titers from baseline was observed as early as day 29 with VIB4. At high doses, the response was approximately 50% by day 85. Using the Emax model, the reduction in RF titers from baseline was VIB4920 showed a statistically significant dose response for reduction (p<0.001 ) (Figure 15B).

[0076] Example 15 - Methods NF-kB reporter assay HEK293 cells expressing NF-kB luciferase reporter (Panomics ) was modified to stably express human full-length CD40R. 4 Cells / well A 96-well poly-D-lysine coated plate (BD Biosciences) was incubated at 100°C for 30 min at 37°C for 1 h. ces) and cultured in the presence or absence of control or CD40L-specific Tn3 for 16 h. MegaCD40L recombinant protein (1.5 μg / ml) was administered at the indicated concentrations for ∼24 h. , Enzo Biosciences) or D1.1 Ju rkat subclone (ATCC) cells. ciferase Assay System (Promega) was used to Luminescence was measured using a traMax M5 plate reader (Molecular Devices). Detected.

[0077] CD86 upregulation assay As approved by the MedImmune Institutional Review Board, Human blood was collected from healthy donors after centrifugation. After centrifugation, peripheral blood mononuclear cells were isolated from the CPT CD40L was isolated from tubes (BD Biosciences) as indicated. 96-well round bottom plate in the presence of specific Tn3 or mAb (clone 5c8) for 16-18 hours In the plates, recombinant megaCD40L (100 ng / ml, Enzo Bios ciences, PBMCs (2.5–5.0 × 10 5 cells / well) were stimulated. Expression of CD86 on CD19+ B cells was assessed using immunocytometry. Antibodies: CD86 (clone 2331, BD Pharmingen), CD19 (clone The antibody was used (HIB19, BD Pharmingen).

[0078] Human B cell assay PBMCs were isolated using MACS cell separation technology (Miltenyi Biotec). negatively selects all B cells using IgG4, which routinely achieves >95% purity Purified peripheral blood B cells were plated in a 96-well plate in complete medium in a final volume of 150 μl. 0.5-1.0 x 10 B cells per well in bottom plates 5 The cells were cultured at a density of 100 μg / ml. B cell experiments The medium used was 10% FCS, penicillin-streptomycin (100 units / ml penicillin cillin, 100 μg / ml streptomycin), 2-mercaptoethanol (55 μ RPMI 164 supplemented with 1 mM L-glutamine (2 mM), and 5 mM HEPES (5 mM). 0 (Invitrogen). At the start of the culture, anti-IgM F(ab')2 ( 5.0μg / ml, Jackson ImmunoResearch Laborato ries) with or without IL-21 (33 ng / ml, PeproTech Inc.) and megaCD40L (1.5 nM, Enzo Biosciences) On the third or fourth day of culture, B cells were stimulated with Cell Titer-Glo Luminescent Assay (Prom B cell proliferation was quantified by measuring ATP using ega. PC differentiation was quantified by flow cytometry. Cells were obtained at time points and CD19+IgD-CD38 hi It was defined as a cell.

[0079] Mouse SRBC immunization model After direct withdrawal from the bottle, 0.2 ml of SRBCs (Colorado SRBCs) were administered by intraperitoneal injection. Do Serum Company) and Balb / c mice (Jackson Lab ratories) were immunized on day 0. Control (30 mg / kg) or CD40L-specific Tn3 (up to 30 mg / kg, as indicated) was administered daily (intravenously) on days 9–13. On day 14, the frequency of germinal center B cells in the spleen was quantified by flow cytometry. GC B cells are expressed as CD19 + B220 + Fas + PNA + were defined as B cells.

[0080] Platelet aggregation assay Human blood was collected from healthy donors and diluted with A, which contains citrate, glucose, and sodium. After centrifugation, 2 / 3 of the platelet-rich plasma was placed in a polypropylene tube. The platelets were transferred to a tube and treated with apyrase (2 U / ml) to prevent platelet activation during the treatment. ) for 10 min. Platelets were pelleted and diluted with modified Tyrode's buffer (1 37 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 5.6 mM glucose , 3.3 mM NaH2PO4, 20 mM HEPES, 0.1% BSA, and pH 7 .4) and resuspended in .

[0081] mAb (h5c8, or negative control antibody) or anti-CD40L Tn3 was administered to hCD40 Immune complexes were isolated by mixing with L (293 Cell Source) at room temperature for 5 minutes. In some experiments, platelets were pretreated with anti-CD32a prior to the addition of IC. The platelet aggregates were preincubated with antibody (IV.3) for 5 min. Meter (Model 700, Chrono-Log, Havertown, PA) Platelet aggregation assays were performed according to the manufacturer's instructions with agitation at 37°C. After mixing the platelets with the agonist, light transmission was monitored for 12-20 min.

[0082] Phase 1a subjects and study design A Phase I randomized study in healthy adults aged 18-49 years, including women of non-childbearing potential A blinded, placebo-controlled trial was conducted (NCT02151110). Safety and tolerability data for 100 mg / kg / day, as well as data accumulated from previous dose cohorts, The clinical trial protocol and the Dose Escalation Committee (DEC) Based on recommendations from the National Center for Clinical Trials (NCT02234661), subjects were divided into seven dose cohorts (3, 1 Patients were randomized to receive either 0, 30, 100, 300, 1000, or 3000 mg, administered sequentially. TDAR was induced in subjects by subcutaneous immunization with 1 mg of KLH in two separate doses. During the screening period 14 days prior to administration of either VIB4920 or placebo, 15 days after the first KLH immunization and administration of either VIB4920 or placebo The second immunization with KLH was then administered to cohort 5 (300 mg), cohort 6 (100 mg), and cohort 7 (100 mg), respectively. All subjects in cohort 1 (1000 mg) and cohort 7 (3000 mg) were completed on day 43. Three interim analyses were performed according to the protocol when

[0083] PK assay of VIB4920 A validated sandwich ELISA method was used to measure V in human K2EDTA plasma. IB4920 was measured in 1×PBS / 0.1% Tween 20 (PBST). A washing step with was performed after each incubation to remove unbound components. Briefly, Nunc microtiter plates were incubated overnight at 2–8°C with 1 μg / mL of The cells were coated with VIB4920 mouse monoclonal antibody (MedImmune). Samples containing data, quality controls (QC), and VIB4920 were Dilute the cells at 1:50 in 0.5% bovine serum albumin (BSA) / PBST before adding them to the medium. The dilution was performed by the method of minimum dilution factor (MRD). After 2 hours of incubation, biotin was added. Pre-conjugated anti-VIB4920 rat antibody (MedImmune) 1μg / mL The mixture was added to the plate and incubated for 1 hour. HRP (GE Healthcare) and SureBlue™ Tetramethylbenzidine (TMB) peroxidase substrate (KPL, Inc.) was used The binding complexes were visualized by incubating the plate with a microplate reader. Color development was stopped with 0.2 M sulfuric acid before analysis at 450 nm. The range was 0.05 to 1.60 μg / mL; samples with values ​​above the quantitative range were pooled. The solution was diluted with diluted K2EDTA plasma to give a concentration within the measurable range of this method.

[0084] Quantification of sCD40L During the screening period, as well as on days 1, 2, 3, 5, 8, 15, and 2 sCD40L concentrations on days 2, 29, 43, 57, 85, and 113 Plasma samples were taken to measure the β-amyloides concentration. Human sCD40L Platinum ELISA kit qualified to ensure accuracy and precision. Total soluble CD4 in human K2EDTA plasma was measured using a ELISA kit (eBioscience). 0L (free sCD40L and sCD40L bound to VIB4920). Then, to ensure results are comparable and consistent, standards, QC, and Samples containing sCD40L were cultured in 0.5% BSA / PBST and VIB4920-containing uptake The cells were diluted 1:50 in Seiy's diluent according to the MRD method. A washing step with PBST was performed for each infusion. After incubation, unbound components were removed. The diluted samples were then incubated with 100 mM NaCl. The cells were added to plates precoated with sCD40L antibody and incubated for 1.5 hours. Then, HRP-conjugated anti-human sCD40L was added, and the sCD40L captured by the coated antibody was The CD40L was bound to the antibody. Before the analysis, TMB peroxidase substrate and stop solution (phosphate) were added sequentially. The binding complex was visualized by the quantitative range of 6.25–400.00 ng / mL. samples with measurements above the quantitative range were diluted with pooled K2EDTA plasma and The concentration was within the measurable range of this method.

[0085] Measurement of ADA A validated sandwich ELISA method was used to measure the β-K+ / β-K+ / β- ... The presence of ADA against VIB4920 was measured, where a washing step with PBST was After each incubation, unbound components were removed. C and samples were diluted with 1:60 MRD in assay diluent containing 0.5% BSA / PBST. Protein G was diluted and washed using the Pierce™ Protein G coated plate ( ThermoFisher) and incubated for 2 hours in assay diluent. The prepared biotin-labeled VIB4920 was incubated overnight at 1 μg / mL. ADA against VIB4920 was specifically detected by streptavidin. Conjugated HRP (GE Healthcare) and SureBlueTMB peptide By successive incubation with peroxidase substrate (KPL, Inc.), Binding complexes were visualized prior to analysis at 450 nm in a microplate reader. Color development was stopped with 0.2M sulfuric acid. Each sample was subjected to a three-step process. In this case, the sample response is first compared to a statistically determined cutoff OD value, above which below which a sample is considered likely to be ADA positive and below which a sample is considered likely to be ADA negative. Potentially positive samples are then run a second time in the presence of good VIB4920. In other words, inhibition rates equal to or greater than the statistically determined confirmation cut point were used for competitive evaluation. Samples with a confirmatory cup were defined as confirmed positive and were evaluated for titer. Samples below the threshold were considered negative for ADA to VIB4920. Titer samples in pooled human K2EDTA plasma were run until the titer was below the screening cutoff. Serial dilutions were made and the titer results were compared to the highest dilution at which the sample tested positive before testing negative. The results are reported as the reciprocal of

[0086] Evaluation of anti-KLH antibodies Anti-Keyhole antibodies in human serum were detected using a validated sandwich ELISA method. Limpet hemocyanin (KLH) IgG antibodies were measured, and the antibody titer was determined by washing with PBST. A washing step was performed after each incubation to remove unbound components. In summary, a volume of 100 μL / well was used in each step. The rates were incubated overnight at 2–8 °C with 3 µg / mL K prepared in 1x PBS (pH 7.2). At LH (Immucothel, biosyn Arzneimittel GmbH) Nine monoclonal anti-KLH antibodies with various isotypes and affinities were coated onto the Standards and QC consist of a mixture of IgG antibodies (AstraZeneca) , and the sample containing anti-KLH antibody were diluted with 0.5% BSA / PBS before being added to the plate. The antibody was diluted 1:250 in T according to the MRD method. After 2 hours of incubation, HRP Conjugated mouse anti-human IgG (Invitrogen) was added to the plate and incubated for 1 hour. The anti-KLH IgG antibody was specifically detected by blotting. Before analysis at 10 nm, TMB peroxidase substrate and stop solution (0.2 M sulfuric acid) were added. The binding complex was visualized by sequential addition of 163.30–10 The measured values ​​were 000.00 ng / mL. Samples with values ​​above the quantitative range were diluted with serum. The concentration was within the measurable range of this method.

[0087] Flow cytometry in Ph1a Blood was collected in a Cytochex BCT tube (Streck) and then analyzed by Covance Central Laboratory Services(Indianapoli) s, IN) and analyzed by flow cytometry using a validated method. In summary, CD45 (clone HI30), CD19 (clone HIB19 ), IgD (clone IA6-2), CD27 (clone M-T271), and CD38 (clone HIT2, all from BD) and cells were stained with fluorochrome-conjugated antibodies to B cell aggregation. The cells were then subjected to FACSPerm2 (Becton Dickinson ) and stained for the expression of Ki67 (clone KI67, Biolegend) in cells. The number of proliferating cells was measured.

[0088] PC Signature PC gene signatures were measured as previously described (Streicher 2014). In summary, the PAXgene Blood RNA kit (Qiagen) was used Total RNA was extracted from PAXgene blood collection tubes using SuperScript II. I First-Strand Synthesis SuperMix Kit (Lif TaqMan q PCR was performed using TaqMan Technologies and random primers. Generate cDNA for PCR. TaqMan Pre-Amp Master Mix Prepare samples using the kit and run them on the BioMark Real-Time PCR System. The inventors analyzed two reference genes (β-actin and GAPDH), ΔΔCt values ​​were calculated using the mean baseline expression levels of each patient as a control. Fold change values ​​were determined by calculating 2-ΔΔCt.

[0089] Phase 1a patients and study design Patients who met the EULAR / ACR criteria (Aletaha) for at least 6 months prior to study entry Phase I included patients aged 18–70 years who were diagnosed with RA according to the This was a phase b randomized, blinded, placebo-controlled trial. DAS28-CRP score of at least 3.2 and at least Moderate to severe, as defined by at least 4 swollen and 4 tender joints Patients had rheumatoid factor-Ig (RF-Ig)-positive rheumatoid arthritis at screening. M ≥ 14 units / mL) or anti-citrullinated peptide antibodies (ACPA) positive Patients were treated with methotrexate (MTX) at doses ranging from 7.5 to 25 mg per week. or in the case of MTX intolerance, at least 12 weeks prior to screening, and A different conventional DMARD was started at a stable dose for at least 6 weeks prior. The biological effect of RA on the Prior treatment with targeted agents (excluding rituximab or other B-cell depleting agents) was permitted. Placebo (n=15) or VIB4920 (75 mg) administered intravenously every other week for 12 weeks. n=8; 500mg n=10; 1000mg n=12; or 1500mg n=1 2) Patients were treated with VECTRA-DA and then followed up for 12 weeks after treatment. The measurements were performed at Crescendo Bioscience (San Francisco, CA). The study was performed by Covance Central Lab (CA) and RF autoantibody measurements were performed by Covance Central Lab (CA). Oratories Services (Princeton, NJ) .

[0090] statistical analysis Primary human B cell expansion in the SRBC model was evaluated using a two-tailed unpaired Student's t-test. The effects of treatment on proliferation, plasma differentiation, and GC B cell responses were evaluated. -U test was used to evaluate VIB4920 at multiple time points for gene signature scores. Statistical tests and plots were performed using Graphpad Pro. The study was carried out using the sm software. Three candidate models were prespecified for the dose-response (linear , Emax, and Hill-Emax models) with the corresponding baseline as a covariate. Using the MCP-Mod method, which includes The dose-response for the change from the line was analyzed. The dose-response signal was examined for multiplicity. The family-wise error rate was controlled at the 0.10 level by adjusting the AIC. Based on the above, the final model was selected from the models that were shown to be significant. Mixed-effects model for repeated measures (MMRM) analyses were performed with baseline outcome included as a covariate. Use it to measure DAS28-CRP, RF, Vectra DA, CDAI, tender joint count, and swelling. Changes in baseline in swollen joint counts, patient and physician global assessments, and serum CRP The changes were analyzed.

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Claims

1. 1. A composition for use in a method for treating or preventing an autoimmune disease or disorder in a subject, the composition comprising a CD40L-specific Tn3 scaffold protein, the method comprising administering to the subject a dose of about 3000 mg of the CD40L-specific Tn3 scaffold protein, the CD40L-specific Tn3 scaffold protein comprising two monomer sequences and a human serum albumin sequence, each monomer sequence comprising the polypeptide sequence of SEQ ID NO: 3, and the autoimmune disease or disorder is one of focal segmental glomerulosclerosis, systemic sclerosis, myositis, antiphospholipid syndrome, autoimmune hepatitis, lupus nephritis, idiopathic thrombocytopenic purpura, vasculitis, cutaneous lupus, autoimmune hemolytic anemia, IgG4-related disease, or systemic lupus erythematosus.

2. 1. A composition for use in a method for treating or preventing an autoimmune disease or disorder in a subject, the composition comprising a CD40L-specific Tn3 scaffold protein, the method comprising administering to the subject the CD40L-specific Tn3 scaffold protein at a dose of about 3000 mg, the CD40L-specific Tn3 scaffold protein comprising SEQ ID NO:1, and the autoimmune disease or disorder is one of focal segmental glomerulosclerosis, systemic sclerosis, myositis, antiphospholipid syndrome, autoimmune hepatitis, lupus nephritis, idiopathic thrombocytopenic purpura, vasculitis, cutaneous lupus, autoimmune hemolytic anemia, IgG4-related disease, or systemic lupus erythematosus.

3. 3. The composition of claim 1, wherein the dose is administered every 14 days or twice a month.

4. The composition of any one of claims 1 to 3, wherein the administration is intravenous.

5. The composition of any one of claims 1 to 4, wherein said administration is effective to reduce levels of an inflammatory marker.

6. 6. The composition of claim 5, wherein the inflammatory markers comprise one or more of autoantibody levels, plasma cell (PC) signature, circulating B cells, and antibody class switching.

7. The composition of claim 6 , wherein the reduction in the level of the inflammatory marker comprises a reduction in the PC signature.

8. 8. The composition of claim 7, wherein the decreased PC signature is characterized by decreased expression of genes selected from the group consisting of immunoglobulin heavy chain constant region alpha 1 (IGHA1), polymeric IgA and IgM binding chain (IGJ), immunoglobulin kappa constant (IGKC), immunoglobulin kappa variable 4-1 (IGKV4-1) and TNF receptor superfamily member 17 (TNFRSF17).

9. 1. A composition for use in a method for treating or preventing an autoimmune disease or disorder in a subject, the composition comprising a CD40L-specific Tn3 scaffold protein, the method comprising administering the CD40L-specific Tn3 scaffold protein to a subject at a dose of about 3000 mg, the CD40L-specific Tn3 scaffold protein comprising two monomer sequences and a human serum albumin sequence, each monomer sequence comprising the polypeptide sequence of SEQ ID NO: 3, and the autoimmune disease or disorder is rheumatoid arthritis.

10. 1. A composition for use in a method for treating or preventing an autoimmune disease or disorder in a subject, the composition comprising a CD40L-specific Tn3 scaffold protein, the method comprising administering the CD40L-specific Tn3 scaffold protein to a subject at a dose of about 3000 mg, the CD40L-specific Tn3 scaffold protein comprising SEQ ID NO:1, and the autoimmune disease or disorder is rheumatoid arthritis.

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