Pharmaceutical composition of anti-CD20 antibody and application thereof

Abstract

The present invention relates to the field of pharmacy and medicine, in particular to pharmaceutical compositions of anti-CD20 antibodies, in particular divozilimab, which compositions can be used for the treatment of CD20-mediated diseases or conditions. The invention further relates to the use of said composition for the treatment of a CD20-mediated disease or disorder, and to a method of treating a CD20-mediated disease or disorder, comprising administering said composition.

Description

Technical Field

[0001] The present invention relates to the fields of pharmacy and medicine, and in particular to a pharmaceutical composition of an anti-CD20 antibody (particularly divozilimab), which can be used to treat CD20-mediated diseases or disorders. Background Art

[0002] Lymphocytes are one of several white blood cell populations; they specifically recognize and respond to foreign antigens. The three main categories of lymphocytes are B lymphocytes (B cells), T lymphocytes (T cells) and natural killer (NK) cells. B lymphocytes are cells responsible for antibody production and humoral immune responses. B cells mature in the bone marrow and leave the bone marrow, expressing antigen-binding antibodies on their cell surfaces. When immature B cells first encounter an antigen specific to membrane-bound antibodies, the cells begin to divide rapidly, and their offspring differentiate into memory B cells and effector cells called "plasma cells". Memory B cells have a long lifespan, and they continue to express membrane-bound antibodies with the same specificity as the original parental B cells. Plasma cells do not produce membrane-bound antibodies, instead they produce secretory forms of antibodies. Secreted antibodies are the main effector molecules of humoral immune responses.

[0003] The CD20 antigen (also known as human B lymphocyte-restricted differentiation antigen, Bp35) is a hydrophobic transmembrane protein with a molecular weight of about 35 kDa located on pre-B lymphocytes and mature B lymphocytes (Valentine et al. J. Biol. Chem. 264 (19): 1989, pp. 11282-11287; and Einfeld et al. EMBO J. 7 (3), 1988, pp. 711-717). The antigen is also expressed on more than 90% of B cell non-Hodgkin lymphomas (NHL) (Anderson et al. Blood 63 (6), 1984, pp. 1424-1433), but is not found in hematopoietic stem cells, pro-B cells, normal plasma cells or other normal tissues (Tedder et al., J. Immunol. 135 (2), 1985, pp. 973-979). CD20 is thought to regulate early steps in the activation process of cell cycle initiation and differentiation (Tedder et al., supra), and may function as a calcium ion channel (Tedder et al., J. Cell. Biochem. 14D, 1990, p. 195).

[0004] The prior art provides anti-CD20 antibodies: rituximab, obinutuzumab, ofatumumab, omezumab, etc.

[0005] In addition, the prior art provides pharmaceutical compositions of various anti-CD20 antibodies. A pharmaceutical composition is known, which comprises rituximab, sodium citrate, polysorbate 80, sodium chloride, hydrochloric acid and sodium hydroxide (Mabthera, a concentrate for a solution for infusion, Russian registration certificate No. N013127 / 01). A pharmaceutical composition is known, which comprises rituximab, recombinant human hyaluronidase, histidine, histidine hydrochloride monohydrate, trehalose dihydrate, methionine, polysorbate 80 (Mabthera, a solution for subcutaneous administration, Russian registration certificate No. (000358)-(PG[RG]-RU)). A pharmaceutical composition is known, which comprises obinutuzumab, histidine, histidine hydrochloride monohydrate, trehalose dihydrate, poloxamer 188 (Gazyva, a concentrate for a solution for infusion, Russian registration certificate No. ЛП[LP]-002867). A pharmaceutical composition is known, which comprises ofatumumab, arginine, sodium acetate trihydrate, sodium chloride, polysorbate 80, EDTA disodium dihydrate, hydrochloric acid (Kesimpta, EMEA / H / C / 005410). A pharmaceutical composition is known, which comprises omezumab, sodium acetate trihydrate, acetic acid, trehalose dihydrate and polysorbate 20 (Okrevus, concentrate for solution for infusion, Russian registration certificate No. ЛП[LP]-004503).

[0006] Divozilimab also relates to anti-CD20 antibodies (WHO Drug Information, Vol. 35, No. 1, 2021, CAS2254061-60-2).

[0007] The prior art does not provide any pharmaceutical composition comprising divozilimab as an active ingredient.

[0008] In view of the above, there is a need to develop improved stable pharmaceutical compositions of anti-CD20 antibodies (particularly divozilimab) which are stable in terms of "monomer content" and "acidic fraction content" during storage and can be used as pharmaceuticals for treating CD20-mediated diseases or disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 is the amino acid sequence of the monoclonal antibody divozilimab showing the number of potential chemical degradation sites for the light and heavy chains.

[0010] Figure 2 is the monomer fraction content relative to time determined by size exclusion HPLC at +37°C.

[0011]

[0012] Figure 3 It is the content of the acidic fraction relative to time determined by ion exchange HPLC at +37°C.

[0013] Formulation 1, Formulation 2 and Formulation 3 have been Figure 2 As mentioned above.

[0014] Figure 4 is the monomer fraction content relative to time determined by size exclusion HPLC at a temperature of +25°C.

[0015] Formulation 1, Formulation 2 and Formulation 3 have been Figure 2 As mentioned above.

[0016] Figure 5 It is the content of the acidic fraction relative to time determined by ion exchange HPLC at a temperature of +25°C.

[0017] Formulation 1, Formulation 2 and Formulation 3 have been Figure 2 As mentioned above.

[0018] Figure 6 is the monomer fraction content relative to time as determined by size exclusion HPLC at a temperature of +(2-8)°C.

[0019] Formulation 1, Formulation 2 and Formulation 3 have been Figure 2 As mentioned above.

[0020] Figure 7 It is the content of the acidic fraction relative to time measured by ion exchange HPLC at a temperature of +(2-8)°C.

[0021] Formulation 1, Formulation 2 and Formulation 3 have been Figure 2 As mentioned above. DETAILED DESCRIPTION

[0022] definition

[0023] Unless otherwise defined herein, all technical and scientific terms used in connection with the present invention shall have the same meanings as commonly understood by one of ordinary skill in the art.

[0024] Furthermore, unless otherwise required by context, singular terms shall include plural terms and plural terms shall include the singular.

[0025] As used in this specification and the appended claims, unless the context requires otherwise, the words “have”, “include” and “comprise” or variations such as “has”, “having”, “includes”, “including”, “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0026] The term "antibody" or "immunoglobulin (Ig)" includes full-length antibodies or any antigen-binding fragment (i.e., "antigen-binding portion") or individual chains thereof. The term "antibody" within the scope of the present invention is used in the broadest sense and may include, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, humanized antibodies, fully human antibodies, and chimeric antibodies.

[0027] Full-length antibody refers to a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated as VH in this specification) and a heavy chain constant region. The constant region is the same in all antibodies of the same isotype, but is different in antibodies of different isotypes. Heavy chains γ, α and δ have a constant region consisting of three constant domains CH1, СH2 and CH3 (on a line), and a hinge region for increasing flexibility (Woof J., Burton D., Nat Rev Immunol 4, 2004, pages 89-99). In mammals, only two types of light chains are known, represented by λ and κ. Each light chain consists of a light chain variable region (abbreviated as VL in this specification) and a light chain constant region. The approximate length of the light chain is 211-217 amino acids. Preferably, the light chain is a λ light chain, and the constant domain CL is preferably Cλ.

[0028] The VL and VH regions can be further subdivided into highly variable regions called complementarity determining regions (CDRs), located between more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0029] As used in this specification, the term "antigen binding portion" or "antigen binding fragment" of an antibody refers to one or more antibody fragments that retain the ability to specifically bind to an antigen. It has been shown that the antigen binding function of an antibody can be performed by a fragment of a full-length antibody. As used in the present invention, the term "antigen binding fragment" refers to a Fab fragment, i.e., a monovalent fragment consisting of VL, VH, CL and CH1 domains, which is connected to an Fc fragment monomer.

[0030] An antibody that "binds" to a target antigen of the present invention refers to an antibody that binds to the antigen with sufficient affinity so that the antibody can be used as a diagnostic and / or therapeutic agent targeting a protein or a cell or tissue expressing the antigen, and slightly cross-reacts with other proteins. Depending on the analytical method: fluorescence activated cell sorting (FACS), radioimmunoassay (RIA) or ELISA, in such an embodiment, the extent of binding of the antibody to non-target proteins is less than 10% of the binding of the antibody to the specific target protein. With respect to the binding of an antibody to a target molecule, the term "specific binding" or the phrase "specific binding" or "specific for a specific polypeptide or an epitope on a specific target polypeptide" refers to binding that is significantly (measurably) different from non-specific interactions.

[0031] The terms "anti-CD20 antibody", "antibody to CD20", "antibody that specifically binds to CD20" or "antibody against CD20" and the like are interchangeable within the framework of the present invention and refer to an antibody that specifically binds to CD20.

[0032] The term "pharmaceutical composition" refers to a composition and / or formulation comprising a therapeutically effective amount of an anti-CD20 antibody and excipients or auxiliary substances (carriers, diluents, fillers, solvents, etc.), the selection and proportion of which depends on the type and route of administration and the dosage.

[0033] The term "excipient" or "auxiliary substance" is used herein to describe any ingredient other than the compounds of the invention. These are substances of inorganic or organic nature used in drug production / manufacturing to impart the necessary physicochemical properties to the drug product.

[0034] As used herein, the term "aqueous composition" refers to a water-based composition, and the water in the composition can be: water, water for injection, physiological saline (0.9%-1.0% sodium chloride aqueous solution).

[0035] As used herein, the term "lyophilized" refers to a preparation that has been subjected to a process known in the art such as freeze-drying, which involves freezing the preparation followed by removing the ice from the frozen contents.

[0036] A pharmaceutical composition is "stable" if the active agent maintains its physical stability and / or chemical stability and / or biological activity at the storage temperature (e.g., 2-8° C.) over a specified shelf life. Additionally, the active agent may maintain both physical and chemical stability, as well as biological activity. The shelf life is adjusted based on the results of stability testing under accelerated aging or natural aging conditions.

[0037] Typically, the amino acid is an L-amino acid. For example, if histidine and histidine hydrochloride monohydrate are used, they are typically L-histidine and L-histidine hydrochloride monohydrate. For example, if proline is used, it is typically L-proline. Amino acid equivalents may also be used, for example, pharmaceutically acceptable proline salts (e.g., proline hydrochloride).

[0038] The term "drug" or "preparation" refers to tablets, capsules, solutions, ointments and other ready-made forms of substances (or mixtures of substances in the form of pharmaceutical compositions) intended for use in restoring, improving or changing the physiological functions of humans and animals, as well as for treating and preventing diseases, for diagnosis, anesthesia, contraception, beauty, etc.

[0039] The term "use" applies to the possibility of using the pharmaceutical composition of the anti-CD20 antibody according to the present invention to treat, alleviate the progress of a disease or condition, accelerate the alleviation of the disease or condition, or reduce the recurrence rate of the disease or condition.

[0040] The term "CD20-mediated disease or disorder" refers to any disease or disorder that is directly or indirectly associated with CD20, including the etiology, development, progression, persistence, or pathology of the disease or disorder.

[0041] "Treat," "treatment," and "therapy" refer to a method of alleviating or eliminating a biological disorder and / or at least one of its associated symptoms. As used herein, "alleviating" a disease, disorder, or condition refers to reducing the severity and / or frequency of the symptoms of the disease, disorder, or condition. In addition, references to "treatment" herein include references to curative, palliative, and preventative treatments.

[0042] The term "parenteral administration" refers to a regimen of administration, usually by injection (infusion), and specifically includes intravenous, intramuscular, intraarterial, intratracheal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection or infusion.

[0043] abbreviation

[0044] VE-vertical electrophoresis

[0045] IE HPLC-ion exchange high performance liquid chromatography.

[0046] SE HPLC-size exclusion high performance liquid chromatography.

[0047] PTM - post-translational modification.

[0048] ABP-acid-base profile.

[0049] N / A - Not applicable.

[0050] The authors of the present invention have found that the presence of trehalose in the pharmaceutical composition of divozilimab negatively affects the stability of the antibody. Furthermore, within the framework of the present invention, it has been determined that the developed pharmaceutical composition has a high stability in terms of "monomer content" and "acidic fraction content".

[0051] The present invention discloses improved stable pharmaceutical compositions of anti-CD20 antibodies, in particular divozilimab, which can be used as medicines for treating CD20-mediated diseases or conditions.

[0052] Within the framework of the present invention, it has been determined that, due to the resulting combination of excipients, the developed pharmaceutical composition has an increased stability with respect to "monomer content" and "acidic fraction content".

[0053] Protein aggregation is defined as the self-association of monomers in a native or partially unfolded form. Many diseases (including Alzheimer's disease, prion disease) are associated with protein aggregation in vivo. Aggregation is also a common manifestation of instability observed in the production, storage and use of biological products. Aggregation may affect efficacy and biodistribution, as well as increase the chance of adverse immune responses in patients. Adverse immune responses to therapeutic proteins have been well documented and may be clinically manifested as reduced efficacy of drugs, infusion reactions, cytokine release syndrome, allergic reactions or even death (Moussa EM, Panchal JP, Moorthy BS, Blum JS, Joubert MK, Narhi LO, Topp EM. Immunogenicity of Therapeutic Protein Aggregates. J Pharm Sci. February 2016).

[0054] According to Ph. Eur. (European Pharmacopoeia), isoform distribution should be analyzed. Anionic modifications that cause a 1-2 unit decrease in pI show reduced plasma clearance and tissue accumulation. (Ryman JT, Meibohm B. Pharmacokinetics of Monoclonal Antibodies. CPT Pharmacometrics Syst Pharmacol. July 2017). These indicators are critical.

[0055] In selecting a formulation, we consider the purpose, route of administration, and tolerability of the drug product (eg, reduced discomfort during administration), as well as the stability and maintenance of activity of the protein molecule within the formulation.

[0056] In one aspect, the present invention relates to a pharmaceutical composition of an anti-CD20 antibody, comprising:

[0057] (i) anti-CD20 antibodies;

[0058] (ii) acetate buffer;

[0059] (iii) sorbitol;

[0060] (iv) methionine;

[0061] (v) Water for injection.

[0062] The concentration of anti-CD20 antibodies included in the pharmaceutical compositions of the present invention can vary depending on the desired properties of the composition and the particular conditions, methods, and intended uses of the pharmaceutical composition.

[0063] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 1.5-120.0 mg / ml.

[0064] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 1.5-1000.0 mg / ml.

[0065] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 1.5-80.0 mg / ml.

[0066] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 1.5-60.0 mg / ml.

[0067] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 5.0-50.0 mg / ml.

[0068] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 10.0-50.0 mg / ml.

[0069] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 10.0-40.0 mg / ml.

[0070] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 15.0-35.0 mg / ml.

[0071] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 20.0-30.0 mg / ml.

[0072] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 22.0-27.0 mg / ml.

[0073] In some embodiments of the invention, the anti-CD20 antibody is present at a concentration of 25.0 mg / ml.

[0074] The anti-CD20 antibody may be an antibody that specifically binds to CD20. The anti-CD20 antibody may be a full-length antibody or an antigen-binding fragment thereof that specifically binds to CD20. The anti-CD20 antibody may have different specificities (e.g., monospecific, bispecific antibodies), different valencies (e.g., monovalent, bivalent, trivalent antibodies), different forms (e.g., classical antibodies, scFv, scFv-Fc, Minibody), and different origins (e.g., mouse, human, camel, chimeric antibodies).

[0075] The antibodies according to the present invention may be of any class (eg, IgA, IgD, IgE, IgG and IgM) or subclass (isotype) (eg, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2).

[0076] The anti-CD20 antibody can be, for example, rituximab, obinutuzumab, ofatumumab, omezumab, and the like.

[0077] In some embodiments of the invention, the anti-CD20 antibody is divozilimab.

[0078] In some embodiments of the invention, the acetate buffer is a mixture of sodium acetate and acetic acid.

[0079] In some embodiments of the invention, sodium acetate is present at a concentration of 0.5-3.0 mg / ml.

[0080] In some embodiments of the invention, sodium acetate is present at a concentration of 1.0-2.5 mg / ml.

[0081] In some embodiments of the invention, sodium acetate is present at a concentration of 1.0-2.0 mg / ml.

[0082] In some embodiments of the invention, sodium acetate is present at a concentration of 1.5-2.0 mg / ml.

[0083] In some embodiments of the invention, sodium acetate is present at a concentration of 1.7-1.8 mg / ml.

[0084] In some embodiments of the invention, sodium acetate is present at a concentration of 1.74 mg / ml.

[0085] In some embodiments of the invention, the sodium acetate is sodium acetate trihydrate.

[0086] In some embodiments of the invention, acetic acid is added to a pH of 5.0-6.0.

[0087] In some embodiments of the invention, acetic acid is added to a pH of 5.4-5.6.

[0088] In some embodiments of the invention, acetic acid is added to pH 5.5.

[0089] In some embodiments of the invention, the acetic acid is glacial acetic acid.

[0090] In some embodiments of the invention, sorbitol is present at a concentration of 40.0-60.0 mg / ml.

[0091] In some embodiments of the invention, sorbitol is present at a concentration of 45.0-55.0 mg / ml.

[0092] In some embodiments of the invention, sorbitol is present at a concentration of 50.0 mg / ml.

[0093] In some embodiments of the invention, methionine is present at a concentration of 0.05-0.30 mg / ml.

[0094] In some embodiments of the invention, methionine is present at a concentration of 0.10-0.20 mg / ml.

[0095] In some embodiments of the invention, methionine is present at a concentration of 0.15 mg / ml.

[0096] In some embodiments of the invention, the pharmaceutical composition comprises:

[0097] (i) anti-CD20 antibodies;

[0098] (ii) an acetate buffer which is a mixture of:

[0099]

[0100] In some embodiments of the invention, the pharmaceutical composition comprises:

[0101] (i) anti-CD20 antibodies;

[0102] (ii) an acetate buffer which is a mixture of:

[0103]

[0104] In some embodiments of the invention, the pharmaceutical composition comprises:

[0105] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0106] (ii) an acetate buffer which is a mixture of:

[0107]

[0108] In some embodiments of the invention, the pharmaceutical composition comprises:

[0109] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0110] (ii) an acetate buffer which is a mixture of:

[0111]

[0112] In some embodiments of the invention, the pharmaceutical composition comprises:

[0113] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0114] (ii) an acetate buffer which is a mixture of:

[0115]

[0116]

[0117] In some embodiments of the invention, the pharmaceutical composition comprises:

[0118] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0119] (ii) an acetate buffer which is a mixture of:

[0120]

[0121] In some embodiments of the invention, the pharmaceutical composition comprises:

[0122] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0123] (ii) an acetate buffer which is a mixture of:

[0124]

[0125] In some embodiments of the invention, the pharmaceutical composition comprises:

[0126] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0127] (ii) an acetate buffer which is a mixture of:

[0128]

[0129] In some embodiments of the invention, the pharmaceutical composition comprises:

[0130] (i) anti-CD20 antibody 25.0 mg / ml;

[0131] (ii) an acetate buffer which is a mixture of:

[0132]

[0133] In some embodiments of the invention, the pharmaceutical composition comprises:

[0134] (i) anti-CD20 antibody 25.0 mg / ml;

[0135] (ii) an acetate buffer which is a mixture of:

[0136]

[0137] In some embodiments of the invention, the pharmaceutical composition comprises:

[0138] (i) anti-CD20 antibodies;

[0139] (ii) an acetate buffer which is a mixture of:

[0140]

[0141] In some embodiments of the invention, the pharmaceutical composition comprises:

[0142] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0143] (ii) an acetate buffer which is a mixture of:

[0144]

[0145]

[0146] In some embodiments of the invention, the pharmaceutical composition comprises:

[0147] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0148] (ii) an acetate buffer which is a mixture of:

[0149]

[0150] In some embodiments of the invention, the pharmaceutical composition comprises:

[0151] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0152] (ii) an acetate buffer which is a mixture of:

[0153]

[0154] In some embodiments of the invention, the pharmaceutical composition comprises:

[0155] (i) anti-CD20 antibody 25.0 mg / ml;

[0156] (ii) an acetate buffer which is a mixture of:

[0157]

[0158] In some embodiments of the invention, the pharmaceutical composition comprises:

[0159] (ii) anti-CD20 antibodies;

[0160] (iii) acetate buffer which is a mixture of:

[0161]

[0162] In some embodiments of the invention, the pharmaceutical composition comprises:

[0163] (i) anti-CD20 antibodies;

[0164] (ii) an acetate buffer which is a mixture of:

[0165]

[0166] In some embodiments of the invention, the pharmaceutical composition comprises:

[0167] (ii) anti-CD20 antibody 1.5-120.0 mg / ml;

[0168] (iii) acetate buffer which is a mixture of:

[0169]

[0170] In some embodiments of the invention, the pharmaceutical composition comprises:

[0171] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0172] (ii) an acetate buffer which is a mixture of:

[0173]

[0174]

[0175] In some embodiments of the invention, the pharmaceutical composition comprises:

[0176] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0177] (ii) an acetate buffer which is a mixture of:

[0178]

[0179] In some embodiments of the invention, the pharmaceutical composition comprises:

[0180] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0181] (ii) an acetate buffer which is a mixture of:

[0182]

[0183] In some embodiments of the invention, the pharmaceutical composition comprises:

[0184] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0185] (ii) an acetate buffer which is a mixture of:

[0186]

[0187] In some embodiments of the invention, the pharmaceutical composition comprises:

[0188] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0189] (ii) an acetate buffer which is a mixture of:

[0190]

[0191] In some embodiments of the invention, the pharmaceutical composition comprises:

[0192] (i) anti-CD20 antibody 25.0 mg / ml;

[0193] (ii) an acetate buffer which is a mixture of:

[0194]

[0195] In some embodiments of the invention, the pharmaceutical composition comprises:

[0196] (i) anti-CD20 antibody 25.0 mg / ml;

[0197] (ii) an acetate buffer which is a mixture of:

[0198]

[0199] In some embodiments of the invention, the pharmaceutical composition comprises:

[0200] (i) anti-CD20 antibodies;

[0201] (ii) an acetate buffer which is a mixture of:

[0202]

[0203]

[0204] In some embodiments of the invention, the pharmaceutical composition comprises:

[0205] (i) anti-CD20 antibodies;

[0206] (ii) an acetate buffer which is a mixture of:

[0207]

[0208] In some embodiments of the invention, the pharmaceutical composition comprises:

[0209] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0210] (ii) an acetate buffer which is a mixture of:

[0211]

[0212] In some embodiments of the invention, the pharmaceutical composition comprises:

[0213] (i) anti-CD20 antibody 1.5-120.0 mg / ml;

[0214] (ii) an acetate buffer which is a mixture of:

[0215]

[0216] In some embodiments of the invention, the pharmaceutical composition comprises:

[0217] (i) anti-CD20 antibody 1.5-60.0 mg / ml;

[0218] (ii) an acetate buffer which is a mixture of:

[0219]

[0220] In some embodiments of the invention, the pharmaceutical composition comprises:

[0221] (i) anti-CD20 antibody 1.5-60.0 mg / ml;

[0222] (ii) an acetate buffer which is a mixture of:

[0223]

[0224] In some embodiments of the invention, the pharmaceutical composition comprises:

[0225] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0226] (ii) an acetate buffer which is a mixture of:

[0227]

[0228] In some embodiments of the invention, the pharmaceutical composition comprises:

[0229] (i) anti-CD20 antibody 5.0-50.0 mg / ml;

[0230] (ii) an acetate buffer which is a mixture of:

[0231]

[0232]

[0233] In some embodiments of the invention, the pharmaceutical composition comprises:

[0234] (i) anti-CD20 antibody 10.0-40.0 mg / ml;

[0235] (ii) an acetate buffer which is a mixture of:

[0236]

[0237] In some embodiments of the invention, the pharmaceutical composition comprises:

[0238] (i) anti-CD20 antibody 10.0-40.0 mg / ml;

[0239] (ii) an acetate buffer which is a mixture of:

[0240]

[0241] In some embodiments of the invention, the pharmaceutical composition comprises:

[0242] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0243] (ii) an acetate buffer which is a mixture of:

[0244]

[0245] In some embodiments of the invention, the pharmaceutical composition comprises:

[0246] (i) anti-CD20 antibody 20.0-30.0 mg / ml;

[0247] (ii) an acetate buffer which is a mixture of:

[0248]

[0249] In some embodiments of the invention, the pharmaceutical composition comprises:

[0250] (i) anti-CD20 antibody 25.0 mg / ml;

[0251] (ii) an acetate buffer which is a mixture of:

[0252]

[0253] In some embodiments of the invention, the pharmaceutical composition comprises:

[0254] (i) divozilimab 25.0 mg / ml;

[0255] (ii) an acetate buffer which is a mixture of:

[0256]

[0257] In some embodiments of the invention, the pharmaceutical composition comprises:

[0258] (i) anti-CD20 antibody 25.0 mg / ml;

[0259] (ii) an acetate buffer which is a mixture of:

[0260]

[0261]

[0262] In some embodiments of the invention, the pharmaceutical composition comprises:

[0263] (i) divozilimab 25.0 mg / ml;

[0264] (ii) an acetate buffer which is a mixture of:

[0265]

[0266] In one aspect, the present invention relates to a pharmaceutical composition of an anti-CD20 antibody, which is provided in a dry (i.e., powder or granular) form for reconstitution in a suitable solvent (e.g., water) prior to administration. Such a formulation can be prepared, for example, by lyophilization (i.e., a process known in the art as freeze drying), and it includes freezing the product and then removing the solvent from the frozen material.

[0267] In one aspect, the present invention relates to a pharmaceutical composition of an anti-CD20 antibody produced by lyophilizing any of the above pharmaceutical compositions of an anti-CD20 antibody. Thus, the pharmaceutical composition according to the present invention may be an aqueous pharmaceutical composition or a lyophilized pharmaceutical composition (lyophilisate).

[0268] Lyophilisates are used to produce other dosage forms. For example, lyophilisates for producing injectable solutions, lyophilisates for producing concentrates for injectable solutions. Lyophilisates are reconstituted by dissolving them in a suitable solvent (most typically, water for injection). In addition, the lyophilized composition is first reconstituted in a desired volume of solvent (most typically in water), and then further diluted in a suitable solvent (e.g., 5% glucose solution, 0.9% sodium chloride solution).

[0269] The pharmaceutical composition according to the present invention is generally suitable for parenteral administration as a sterile preparation, which is intended to be administered in the human body by bypassing the gastrointestinal tract through a breach in the skin or mucosal barrier by means of injection, infusion and implantation. In particular, parenteral administration is contemplated to include, inter alia, subcutaneous, intraperitoneal, intramuscular, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, ​​intracranial, intrasynovial, transcutaneous injection or infusion; and renal dialysis infusion techniques. Preferred embodiments include intravenous and subcutaneous routes. Any method accepted in the art for administering a peptide or protein may be applicable to the composition of an anti-CD20 antibody according to the present invention.

[0270] The pharmaceutical composition of the anti-CD20 antibody according to the present invention can be used after dilution. To this end, the desired volume of the composition is transferred from a vial to an infusion container containing a sterile 0.9% sodium chloride solution or a sterile 5% glucose solution. The resulting solution is stirred by gently turning the infusion container.

[0271] The pharmaceutical composition according to the present invention can be stored in any suitable container. For example, a glass or plastic container, vial, ampoule, syringe, cartridge or bottle of the desired volume. The container can be provided with additional devices for administration, such as a dropper, automatic syringe.

[0272] The pharmaceutical composition according to the present invention can be manufactured, packaged or widely sold in the form of ready-made preparations in the form of a single unit dose or multiple single unit doses. As used herein, the term "single unit dose" refers to a discrete amount of a pharmaceutical composition containing a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dosage of the active ingredient to be used in the subject, or a convenient portion of such dosage, such as half or one-third of such dosage.

[0273] In one aspect, the present invention relates to the use of the above-mentioned pharmaceutical composition of anti-CD20 antibodies for treating a CD20-mediated disease or disorder in a subject in need thereof.

[0274] In some embodiments of the invention, the pharmaceutical composition is administered at a dose of 125 mg of the anti-CD20 antibody.

[0275] In some embodiments of the invention, the pharmaceutical composition is administered at a dose of 500 mg of the anti-CD20 antibody.

[0276] In some embodiments of the invention, the pharmaceutical composition is administered every 24 weeks.

[0277] In some embodiments of the invention, the pharmaceutical composition is administered every 24 weeks at a dose of 125 mg of the anti-CD20 antibody.

[0278] In some embodiments of the invention, the pharmaceutical composition is administered every 24 weeks at a dose of 500 mg of the anti-CD20 antibody.

[0279] In some embodiments of the present invention, use of the above-described pharmaceutical composition of an anti-CD20 antibody for treating a CD20-mediated disease or disorder in a subject in need thereof comprises administering the pharmaceutical composition for at least 5 cycles.

[0280] In some embodiments of the present invention, a first dose of 125 mg or 500 mg of an anti-CD20 antibody can be administered to a subject at a dose of 62.5 mg or 250 mg in two steps at an interval of 14 days (i.e., on day 1, the anti-CD20 antibody is administered to the subject at a dose of 62.5 mg or 250 mg; on day 15, the anti-CD20 antibody is administered again at a similar dose of 62.5 mg or 250.0 mg). The anti-CD20 antibody is administered in the form of the above-mentioned pharmaceutical composition.

[0281] In some embodiments of the present invention, the use of the above-mentioned pharmaceutical composition of anti-CD20 antibodies for treating CD20-mediated diseases or conditions in subjects in need thereof comprises administering the pharmaceutical composition for at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of the anti-CD20 antibody and a second administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of the anti-CD20 antibody 2 weeks after the first administration; and the subsequent cycles comprise administering the above-mentioned pharmaceutical composition at a dose of 125 mg of the anti-CD20 antibody every 24 weeks after the first administration.

[0282] In some embodiments of the present invention, the use of the above-mentioned pharmaceutical composition of anti-CD20 antibodies for treating CD20-mediated diseases or conditions in subjects in need thereof comprises administering the pharmaceutical composition for at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of the anti-CD20 antibody and a second administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of the anti-CD20 antibody 2 weeks after the first administration; and the subsequent cycles comprise administering the above-mentioned pharmaceutical composition at a dose of 500 mg of the anti-CD20 antibody every 24 weeks after the first administration.

[0283] In some embodiments of the invention, the anti-CD20 antibody is divozilimab.

[0284] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 125 mg of divozilimab.

[0285] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 500 mg of divozilimab.

[0286] In some embodiments of the present invention, the above-described pharmaceutical composition of divozilimab is administered every 24 weeks.

[0287] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 125 mg of divozilimab every 24 weeks.

[0288] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 500 mg of divozilimab every 24 weeks.

[0289] In some embodiments of the present invention, the use of the above-mentioned pharmaceutical composition of divozilimab for treating a CD20-mediated disease or disorder in a subject in need thereof comprises administering at least 5 cycles of the above-mentioned pharmaceutical composition of divozilimab.

[0290] In some embodiments of the present invention, the use of the above-mentioned pharmaceutical composition of divozilimab for treating a CD20-mediated disease or condition in a subject in need thereof comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of divozilimab and a second administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of divozilimab 2 weeks after the first administration; and subsequent cycles comprise administering the above-mentioned pharmaceutical composition at a dose of 125 mg of divozilimab every 24 weeks after the first administration.

[0291] In some embodiments of the present invention, the use of the above-mentioned pharmaceutical composition of divozilimab for treating a CD20-mediated disease or condition in a subject in need thereof comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of divozilimab and a second administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of divozilimab 2 weeks after the first administration; and subsequent cycles comprise administration of the above-mentioned pharmaceutical composition at a dose of 500 mg of divozilimab every 24 weeks after the first administration.

[0292] In some embodiments of the invention, the CD20-mediated disease or disorder is selected from:

[0293] (i) a neoplastic disease or condition, or

[0294] (ii) Autoimmune diseases or disorders.

[0295] In some embodiments of the invention, the neoplastic disease or disorder is selected from the group consisting of: B-cell lymphoma, leukemia.

[0296] In some embodiments of the invention, the B cell lymphoma is non-Hodgkin lymphoma, Hodgkin lymphoma.

[0297] In some embodiments of the invention, the leukemia is selected from the group consisting of: chronic lymphocytic leukemia, small lymphocytic lymphoma.

[0298] In some embodiments of the invention, the autoimmune disease or disorder is selected from the group consisting of: multiple sclerosis, axonal neuropathy, ANCA-associated vasculitis, IgG4-associated disease, amyloidosis, axial spondyloarthritis, antiphospholipid syndrome, Takayasu's arteritis, autoimmune aplastic anemia, autoimmune inner ear disease, autoimmune hemolytic anemia, autoimmune hyperlipidemia, autoimmune familial dysautonomia, autoimmune urticaria, autoimmune neuromyotonia (Isaacs syndrome), autoimmune retinopathy, autoimmune thrombocytopenic purpura, warm antibody hemolytic anemia, autoimmune testicular disease, autoimmune angioedema, autoimmune hepatitis, autoimmune diabetes, autoimmune immunodeficiency, autoimmune Immune pericarditis, autoimmune thyroiditis, autoimmune progesterone dermatitis, Addison's disease, Behcet's disease, Graves' disease (diffuse toxic goiter), neuromyelitis optica spectrum (Devic's disease), Castleman's disease, Crohn's disease, Ormond's disease (retroperitoneal fibrosis), Raynaud's disease, Churg-Strauss syndrome, bullous pemphigoid, epidermolysis bullosa, vasculitis, renal vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, cystic dermatosis, giant cell arteritis, vitiligo, lupus nephritis, inflammatory aortic aneurysm, inflammatory bowel disease, congenital heart block, pemphigus vulgaris, dermatitis herpetiformis, pemphigoid gestationis, hypogammaglobulinemia, glomerulonephritis, granulomatosis with polyangiitis (Wegener's syndrome), dermatomyositis, dilatation cardiomyopathy, demyelinating neuropathy, discoid lupus erythematosus, idiopathic recurrent gross hematuria (IgA nephropathy), idiopathic inflammatory myopathy, idiopathic inflammatory pseudotumor, idiopathic thrombocytopenic purpura, idiopathic hypocomplementemic tubulointerstitial nephritis, idiopathic pulmonary fibrosis, diabetes mellitus, pure red cell aplasia, Barlow's concentric sclerosis, lichen planus, lichen sclerosus, leukocytoclastic vasculitis, limited scleroderma (CREST syndrome), limbic encephalitis, linear IgA bullous dermatosis, pemphigus foliaceus, Waldenstrom's macroglobulinemia, mediastinal fibrosis, myasthenic crisis, Lang-Evans myasthenic syndrome, myasthenia gravis, microscopic polyangiitis, myositis, Coxsackie myocarditis, multiple sclerosis, multifocal motor neuropathy, Multifocal fibrosclerosis, optic neuritis, undifferentiated connective tissue disease uveitis, neutropenia, ulcerative colitis, anti-tubular / membrane antibody-associated nephritis, opsoclonus, acute motor axonal neuropathy, acute hemorrhagic leukoencephalitis, acute disseminated encephalomyelitis, relapsing rheumatic disease, paraneoplastic cerebellar degeneration, paraneoplastic syndrome, paraproteinemic polyneuropathy, paroxysmal nocturnal hemoglobinuria, ocular pemphigus, mucous membrane pemphigoid, primary biliary cirrhosis, primary sclerosing cholangitis, periaortitis, perianal arteritis, perivenous encephalomyelitis, peripheral neuropathy, peripheral uveitis, pernicious anemia, subacute bacterial endocarditis, polymyositis, polyneuropathy with organomegaly (POEMS syndrome),Transverse myelitis, post-myocardial infarction syndrome, post-traumatic pericarditis, psoriasis, psoriatic arthritis, pemphigus, Henoch-Schönlein purpura, anti-HLA antibody-mediated transplant rejection, reactive arthritis, rheumatic fever, polymyalgia rheumatoid, rheumatoid arthritis, reflex sympathetic dystrophy, refractory epilepsy, relapsing polychondritis, sarcoidosis, acute febrile polyneuritis, Kawasaki syndrome, Cogan syndrome, Mikulic syndrome, stiff-man syndrome, Sjögren syndrome, Evans syndrome, systemic lupus erythematosus, systemic scleroderma, localized scleroderma, Riedel's thyroiditis, Hashimoto's thyroiditis, thyroid-related eye disease, thyroid Glandular crisis, thrombotic thrombocytopenic purpura, polyarteritis nodosa, fetal and neonatal alloimmune thrombocytopenia, fibrosing alveolitis, B-cell non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia, cold hemagglutinin disease, Sydenham's chorea (small chorea or rheumatic chorea), chronic Lyme disease, chronic inflammatory demyelinating polyneuropathy, chronic relapsing multifocal osteomyelitis, chronic sclerosing sialadenitis (Tutnot tumor), celiac disease, endometriosis, encephalitis, autoantibody-related encephalopathy, Hashimoto's encephalitis, eosinophilic fasciitis, essential mixed cryoglobulinemia, juvenile diabetes, juvenile rheumatoid arthritis.

[0299] In some embodiments of the invention, multiple sclerosis is remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, multiple sclerosis with exacerbations, highly active multiple sclerosis, aggressive multiple sclerosis, malignant multiple sclerosis (Marburg variant), myelin-destructive diffuse Schilder sclerosis, myelocortical multiple sclerosis, clinically isolated syndrome.

[0300] In some embodiments of the present invention, the subject or patient treated is a mammal, preferably a human subject. The subject may be male or female of any age.

[0301] In one aspect, the present invention relates to a method for treating a CD20-mediated disease or disorder, comprising administering the above-mentioned pharmaceutical composition to a subject in need thereof in a therapeutically effective amount.

[0302] In some embodiments of the present invention, the above pharmaceutical composition is administered at a dose of 125 mg of the anti-CD20 antibody.

[0303] In some embodiments of the present invention, the above pharmaceutical composition is administered at a dose of 500 mg of the anti-CD20 antibody.

[0304] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered every 24 weeks.

[0305] In some embodiments of the present invention, the above pharmaceutical composition is administered at a dose of 125 mg of the anti-CD20 antibody every 24 weeks.

[0306] In some embodiments of the present invention, the above pharmaceutical composition is administered at a dose of 500 mg of the anti-CD20 antibody every 24 weeks.

[0307] In some embodiments of the invention, the method of treating a CD20-mediated disease or disorder comprises administering at least 5 cycles of the above-described pharmaceutical composition.

[0308] In some embodiments of the present invention, a first dose of 125 mg or 500 mg of an anti-CD20 antibody can be administered to a subject at a dose of 62.5 mg or 250 mg in two steps at an interval of 14 days (i.e., on day 1, the anti-CD20 antibody is administered to the subject at a dose of 62.5 mg or 250 mg; on day 15, the anti-CD20 antibody is administered again at a similar dose of 62.5 mg or 250.0 mg). The anti-CD20 antibody is administered in the form of the above-mentioned pharmaceutical composition.

[0309] In some embodiments of the present invention, the method for treating a CD20-mediated disease or condition comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody and a second administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody 2 weeks after the first administration; and subsequent cycles comprise administration of the above-mentioned pharmaceutical composition at a dose of 125 mg of an anti-CD20 antibody every 24 weeks after the first administration.

[0310] In some embodiments of the present invention, the method for treating a CD20-mediated disease or condition comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody and a second administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody 2 weeks after the first administration; and subsequent cycles comprise administration of the above-mentioned pharmaceutical composition at a dose of 500 mg of an anti-CD20 antibody every 24 weeks after the first administration.

[0311] In some embodiments of the invention, the anti-CD20 antibody is divozilimab.

[0312] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 125 mg of divozilimab.

[0313] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 500 mg of divozilimab.

[0314] In some embodiments of the present invention, the above-described pharmaceutical composition of divozilimab is administered every 24 weeks.

[0315] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 125 mg of divozilimab every 24 weeks.

[0316] In some embodiments of the present invention, the above-mentioned pharmaceutical composition is administered at a dose of 500 mg of divozilimab every 24 weeks.

[0317] In some embodiments of the present invention, the method of treating a CD20-mediated disease or disorder comprises administering 5 cycles of the above-described pharmaceutical composition of divozilimab.

[0318] In some embodiments of the present invention, the method for treating a CD20-mediated disease or condition comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of divozilimab and a second administration of the above-mentioned pharmaceutical composition at a dose of 62.5 mg of divozilimab 2 weeks after the first administration; and subsequent cycles comprise administering the above-mentioned pharmaceutical composition at a dose of 125 mg of divozilimab every 24 weeks after the first administration.

[0319] In some embodiments of the present invention, the method for treating a CD20-mediated disease or condition comprises at least 5 cycles, wherein cycle 1 comprises a first administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of divozilimab and a second administration of the above-mentioned pharmaceutical composition at a dose of 250 mg of divozilimab 2 weeks after the first administration; and subsequent cycles comprise administration of the above-mentioned pharmaceutical composition at a dose of 500 mg of divozilimab every 24 weeks after the first administration.

[0320] In some embodiments of the invention, the CD20-mediated disease or disorder is selected from:

[0321] (i) a neoplastic disease or condition, or

[0322] (ii) Autoimmune diseases or disorders.

[0323] In some embodiments of the invention, the neoplastic disease or disorder is selected from the group consisting of: B-cell lymphoma, leukemia.

[0324] In some embodiments of the invention, the B cell lymphoma is non-Hodgkin lymphoma, Hodgkin lymphoma.

[0325] In some embodiments of the invention, the leukemia is selected from the group consisting of: chronic lymphocytic leukemia, small lymphocytic lymphoma.

[0326] In some embodiments of the invention, the autoimmune disease or disorder is selected from the group consisting of: multiple sclerosis, axonal neuropathy, ANCA-associated vasculitis, IgG4-associated disease, amyloidosis, axial spondyloarthritis, antiphospholipid syndrome, Takayasu's arteritis, autoimmune aplastic anemia, autoimmune inner ear disease, autoimmune hemolytic anemia, autoimmune hyperlipidemia, autoimmune familial dysautonomia, autoimmune urticaria, autoimmune neuromyotonia (Isaacs syndrome), autoimmune retinopathy, autoimmune thrombocytopenic purpura, warm antibody hemolytic anemia, autoimmune testicular disease, autoimmune angioedema, autoimmune hepatitis, autoimmune diabetes, autoimmune immunodeficiency, autoimmune Immune pericarditis, autoimmune thyroiditis, autoimmune progesterone dermatitis, Addison's disease, Behcet's disease, Graves' disease (diffuse toxic goiter), neuromyelitis optica spectrum (Devic's disease), Castleman's disease, Crohn's disease, Ormond's disease (retroperitoneal fibrosis), Raynaud's disease, Churg-Strauss syndrome, bullous pemphigoid, epidermolysis bullosa, vasculitis, renal vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, cystic dermatosis, giant cell arteritis, vitiligo, lupus nephritis, inflammatory aortic aneurysm, inflammatory bowel disease, congenital heart block, pemphigus vulgaris, dermatitis herpetiformis, pemphigoid gestationis, hypogammaglobulinemia, glomerulonephritis, granulomatosis with polyangiitis (Wegener's syndrome), dermatomyositis, dilatation cardiomyopathy, demyelinating neuropathy, discoid lupus erythematosus, idiopathic recurrent gross hematuria (IgA nephropathy), idiopathic inflammatory myopathy, idiopathic inflammatory pseudotumor, idiopathic thrombocytopenic purpura, idiopathic hypocomplementemic tubulointerstitial nephritis, idiopathic pulmonary fibrosis, diabetes mellitus, pure red cell aplasia, Barlow's concentric sclerosis, lichen planus, lichen sclerosus, leukocytoclastic vasculitis, limited scleroderma (CREST syndrome), limbic encephalitis, linear IgA bullous dermatosis, pemphigus foliaceus, Waldenstrom's macroglobulinemia, mediastinal fibrosis, myasthenic crisis, Lang-Evans myasthenic syndrome, myasthenia gravis, microscopic polyangiitis, myositis, Coxsackie myocarditis, multiple sclerosis, multifocal motor neuropathy, Multifocal fibrosclerosis, optic neuritis, undifferentiated connective tissue disease uveitis, neutropenia, ulcerative colitis, anti-tubular / membrane antibody-associated nephritis, opsoclonus, acute motor axonal neuropathy, acute hemorrhagic leukoencephalitis, acute disseminated encephalomyelitis, relapsing rheumatic disease, paraneoplastic cerebellar degeneration, paraneoplastic syndrome, paraproteinemic polyneuropathy, paroxysmal nocturnal hemoglobinuria, ocular pemphigus, mucous membrane pemphigoid, primary biliary cirrhosis, primary sclerosing cholangitis, periaortitis, perianal arteritis, perivenous encephalomyelitis, peripheral neuropathy, peripheral uveitis, pernicious anemia, subacute bacterial endocarditis, polymyositis, polyneuropathy with organomegaly (POEMS syndrome),Transverse myelitis, post-myocardial infarction syndrome, post-traumatic pericarditis, psoriasis, psoriatic arthritis, pemphigus, Henoch-Schönlein purpura, anti-HLA antibody-mediated transplant rejection, reactive arthritis, rheumatic fever, polymyalgia rheumatoid, rheumatoid arthritis, reflex sympathetic dystrophy, refractory epilepsy, relapsing polychondritis, sarcoidosis, acute febrile polyneuritis, Kawasaki syndrome, Cogan syndrome, Mikulic syndrome, stiff-man syndrome, Sjögren syndrome, Evans syndrome, systemic lupus erythematosus, systemic scleroderma, localized scleroderma, Riedel's thyroiditis, Hashimoto's thyroiditis, thyroid-related eye disease, thyroid Glandular crisis, thrombotic thrombocytopenic purpura, polyarteritis nodosa, fetal and neonatal alloimmune thrombocytopenia, fibrosing alveolitis, B-cell non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia, cold hemagglutinin disease, Sydenham's chorea (small chorea or rheumatic chorea), chronic Lyme disease, chronic inflammatory demyelinating polyneuropathy, chronic relapsing multifocal osteomyelitis, chronic sclerosing sialadenitis (Tutnot tumor), celiac disease, endometriosis, encephalitis, autoantibody-related encephalopathy, Hashimoto's encephalitis, eosinophilic fasciitis, essential mixed cryoglobulinemia, juvenile diabetes, juvenile rheumatoid arthritis.

[0327] In some embodiments of the invention, multiple sclerosis is remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, multiple sclerosis with exacerbations, highly active multiple sclerosis, aggressive multiple sclerosis, malignant multiple sclerosis (Marburg variant), myelin-destructive diffuse Schilder sclerosis, myelocortical multiple sclerosis, clinically isolated syndrome.

[0328] The pharmaceutical composition can be used as a single therapeutic agent or can be used in combination with other therapeutic agents as required. Therefore, in one embodiment, the treatment and / or prevention method of the present invention is used in combination with another activating agent that is used in a therapeutically effective amount. Other activating agents can be used before, during or after the use of the pharmaceutical composition according to the present invention. Other activating agents can be used as a part of the present composition or as a separate preparation.

[0329] Implementation of the Invention

[0330] The following examples are provided for a better understanding of the present invention. These examples are for illustrative purposes only and should not be construed as limiting the scope of the present invention in any way.

[0331] All publications, patents and patent applications cited in this specification are incorporated herein by reference. Although the foregoing invention has been described in considerable detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those skilled in the art based on the teachings of the present invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the attached embodiments.

[0332] method.

[0333] 1. Determination of protein concentration in test samples.

[0334] The protein concentration was determined by UV spectrophotometry at a wavelength of 280 nm in a UV spectrophotometry plate.

[0335] Each sample was diluted to a concentration of about 0.5 mg / ml with an appropriate solution of excipient. 150 μl of the diluted sample was placed in the wells of a UV spectrophotometry plate. The optical density of the solution in the wells was measured using a plate spectrophotometer at a wavelength of 280 nm. The corresponding solution of the excipient was used as a reference solution.

[0336] The protein concentration (C) (mg / ml) was calculated using the following formula:

[0337] С=Α(280)*bε*lС=Α280*bε*l,

[0338] Α280 is the optical density value at a wavelength of 280 nm;

[0339] e is the extinction coefficient of the test protein;

[0340] b is the total dilution factor of the sample;

[0341] l is the layer thickness in the plate well (for 150 μL, l = 0.42 cm; for half-area plates, 175 μL = 1 cm).

[0342] 2. Buffer solution replacement and sample concentration.

[0343] Dialyze and concentrate samples in a variety of ways:

[0344] - Concentrated cells under pressure in Stirred Cells (Millipore),

[0345] - In an Amicon centrifuge tube with a 10 kDa membrane (Millipore),

[0346] - In dialysis tubing with 2 ml or 0.5 ml 10 kDa membrane (Thermo Scientific).

[0347] The dialysis was performed for 2-3 hours, after which the spent dialysis buffer was replaced with fresh dialysis buffer and the dialysis was continued for several more hours with constant stirring in an orbital shaker (100 rpm) at room temperature.

[0348] 3. Evaluate colloidal stability by shaking test.

[0349] The test samples were split into 2 portions of 200 μl each and placed in glass vials, 1 vial per formulation was transferred to a refrigerator and aged at 2-8° C., and the remaining vials were placed in a shaker and shaken at 800 rpm for the specified time at 2-8° C. After stress, the vials were removed from the shaker and transferred for analysis.

[0350] 4. Evaluation of colloidal stability by low temperature concentration.

[0351] The test samples were split into 2 portions and placed in plastic bottles: 1 vial of each formulation was stored in a refrigerator at 2-8°C; the remaining vials were stored in a freezer at minus 16-20°C for the specified period of time. After stress, the vials were removed from the freezer and kept at room temperature until the contents were completely thawed; the solutions were mixed and transferred for analysis.

[0352] 5. Evaluate thermal stability through "thermal stress"

[0353] The test samples were split into 2 portions and placed in separate glass vials: 1 vial of each composition was stored in a refrigerator at 2-8°C and the remaining vial was incubated in a thermostat at the desired temperature for the specified period of time. After heating, the vials were removed from the thermostat, kept at room temperature for approximately 15 minutes, and transferred for analysis.

[0354] 6. Determination of protein homogeneity and aggregation sites by dynamic light scattering (DLS).

[0355] The homogeneity of the test samples was determined using a DynaPro Plate Reader II (Wyatt). To this end, 35 ml of the solution was added to the wells of a 384-well 384LV / EB plate. After filling, the plate was sealed with a membrane. To remove air bubbles from the wells after filling, the plate on a lint-free wipe was centrifuged at 3000 rpm in a plate rotor for 3-5 minutes; during this period, the temperature in the centrifuge chamber should not fall below 20°C to avoid condensation of the sample on the membrane.

[0356] Measurement temperature: 25℃.

[0357] Maintain this temperature for 5 seconds before starting the measurement.

[0358] Scattered light intensity at θ = 158°.

[0359] The number of measurements for each repetition was 10.

[0360] Each measurement lasts 5 seconds.

[0361] Aggregation sites of the test proteins were determined using a DynaPro Plate Reader II (Wyatt). Samples were prepared according to the method described above for determination of homogeneity by DLS.

[0362] The heating temperature range is 40-85°C.

[0363] The heating rate was 0.188°C / min.

[0364] The number of samples per measurement shall not exceed 16.

[0365] Scattered light intensity at θ = 158°.

[0366] The number of measurements for each repetition was 3.

[0367] Each measurement lasts 5 seconds.

[0368] We established the temperature trend using the device software, which also automatically calculated the protein aggregation point (onset temperature).

[0369] 7. The purity of the samples was determined by size exclusion high performance liquid chromatography (SEC HPLC).

[0370] Tosoh column TSK-GelG3000SWXL 7.8mm ID×30cm, accession number 08541.

[0371] Column temperature: 25°С

[0372] Mobile phase flow rate: 0.7ml / min

[0373] Injection volume: 10 μL (sample concentration is 5 mg / ml, diluted with excipient solution). For other concentrations, the injection volume is equal to the concentration.

[0374] Sample concentration: 5 mg / ml.

[0375] Detector wavelength: 220nm.

[0376] Elution time: 25 minutes.

[0377] Mobile phase: anhydrous disodium hydrogen phosphate 7.1 mg / ml.

[0378] Sodium chloride 17.54mg / ml.

[0379] The mobile phase pH was adjusted to 7.0 with orthophosphoric acid.

[0380] 8. The acid-base distribution of the sample was determined on the Caliper LabChip GX II.

[0381] Sample preparation.

[0382] The sample was diluted to a concentration of 1 mg / ml. 2 μl of a 5 mg / ml carboxypeptidase B (CpB) solution was added to 200 μl of the resulting solution. The solution was mixed and incubated at 37°C for 2 hours. The test sample was dialyzed in an Amicon Ultra centrifuge tube with three changes of water and concentrated to 2 mg / ml.

[0383] Preparation of working solutions.

[0384] Using the HT Protein Charge Variant Labeling Kit, prepare working solutions and plates with test samples according to the manufacturer's protocol.

[0385] Chip preparation.

[0386] Prepare the chip and buffer tubes using buffer solutions from the Protein Charge Variant Buffer Kit according to the manufacturer's protocol.

[0387] The assay was initiated following standard procedures. Protein Charge Variant 68s was used as the assay method.

[0388] 9. The acid-base distribution of the samples was determined by ion exchange (IE) HPLC.

[0389] Chromatographic conditions:

[0390] Column: Pro PACΤΜWCX-10Analytical (Dionex), 250×4mm;

[0391] Front column: Pro Pac WCX-10G, 4×50mm;

[0392] Mobile Phase:

[0393] Solution A 0.02 M 2-(N-morpholino)ethanesulfonic acid (MES), pH 6.0;

[0394] Solution B: 0.02 M MES, 0.4 M NaCl, pH 6.0;

[0395] Mobile phase speed: 0.7 ml / min;

[0396] Column temperature: 30°C;

[0397] Autosampler temperature: 5°C;

[0398] Detector: UV, 280nm;

[0399] Injected sample volume: 40 μL.

[0400] 10.PTM bioinformatics.

[0401] Protein structures may contain certain regions that are susceptible to chemical degradation, such as acetylation, carbamylation, methylation, phosphorylation, oxidation, and other processes. The type of chemical modification is determined by the structure of the amino acids in the polypeptide chain or their sequence. In order to predict the propensity of a protein to the above modifications, PTM sites and their number can be bioinformatically studied in the primary structure of the protein.

[0402] 11. Stability study.

[0403] The test samples were divided into several aliquots and placed in different sterile glass vials: 1 vial for each control point was placed in an incubator and incubated at 25°C / 37°С for 6 months / 4 weeks and at (5±3)°C for 24 months, with control points collected periodically according to the schedule. After the control points were collected and stored, the vials were removed from the incubator or refrigerator and transferred for analysis.

[0404] 12. Purity was determined by reducing and non-reducing vertical gel electrophoresis in polyacrylamide gels (PAGs)

[0405] The assay was performed by vertical electrophoresis in polyacrylamide gel (PAAG) in the presence of sodium dodecyl sulfate under reducing and non-reducing conditions (10 and 40 μg load) followed by gel staining with acid blue dye 83.

[0406] The assay was performed using an electrophoresis apparatus with the following parameters: gel width of at least 13 cm, height (from the lower edge of the comb teeth) of at least 12 cm, thickness of 1 mm. To form the wells in the stacking gel, we used a comb with at least 20 teeth, dimensions: width 4-6.5 mm, thickness 1 mm.

[0407] The substance solution was diluted with water to a concentration of 2 mg / ml.

[0408] 80 μl of the resulting solution was mixed with 80 ml of a sample application buffer solution containing 2-mercaptoethanol (reducing conditions) or not containing 2-mercaptoethanol (non-reducing conditions). The resulting mixture was incubated at 100° C. for 1 minute (samples without 2-mercaptoethanol) or 3 minutes (samples containing 2-mercaptoethanol), cooled to room temperature, then centrifuged for 4-6 seconds, and mixed (test solution).

[0409] Reducing electrophoresis was performed in 12.5% ​​PAAG, and non-reducing electrophoresis was performed in 8% PAAG.

[0410] The lower reservoir of the electrophoresis instrument is filled with electrode buffer solution and the electrophoresis cell with gel is inserted into the chamber. The upper reservoir is filled with electrode buffer solution and air bubbles are removed from the wells.

[0411] Reducing and non-reducing electrophoresis were performed at room temperature in constant voltage mode. The voltage was 100 V while the bromophenol blue dye front migrated through the stacking gel. After the bromophenol blue dye front entered 5-7 mm into the lower resolving gel, the voltage was increased to 180 V. After the blue bromophenol dye reached 10 cm from the lower edge of the stacking gel, the voltage was turned off and the gel was separated from the pool glass.

[0412] After non-reducing electrophoresis, the gel is incubated with the fixing solution for 16-18 hours. The solution is then drained, the staining solution is added, and the mixture is incubated on an orbital shaker at 50 rpm for 1 hour. The staining solution is then drained, the washing solution is heated to a temperature of 40-50°C, the gel is poured, and the mixture is placed on an orbital shaker at 50 rpm for 30 minutes. The washing procedure is repeated until the bands in the sensitivity solution track are clearly visible. The stained gel is scanned.

[0413] The number of major diffuse bands observed in the gel after reducing and non-reducing electrophoresis was determined by the traces of solutions containing 10 μg of protein.

[0414] Material purity was assessed using GelPro software by tracing a solution containing 40 μg of protein in sample application buffer with or without 2-mercaptoethanol.

[0415] Example

[0416] The following examples are given for the anti-CD20 antibody divozilimab.

[0417] Example 1.

[0418] The key parameters of acid-base distribution and aggregate content are affected by excipient formulation. This example studies the effect of stabilizers and osmotic agents on the test quality indicators.

[0419] For the study, we used osmolytes at concentrations that provided osmotic pressure in the bloodstream: 270-330 mOsm. The following excipients were screened: sodium acetate trihydrate (buffer, maintains desired pH level), glacial acetic acid (maintains desired pH level), D-trehalose dihydrate (stabilizer, osmotic agent), D-sorbitol (stabilizer, osmotic agent), D-mannitol (stabilizer, osmotic agent), L-proline (stabilizer, osmotic agent).

[0420] The excipient formulations of the test pharmaceutical compositions are shown in Table 1.

[0421] Table 1 - Test drug compositions

[0422]

[0423] Samples containing 5 mg / ml divozilimab were studied. Samples with various excipient formulations were prepared by diafiltration according to Method 2. To identify the effects of various excipients, samples in the test formulations were subjected to stress according to Methods 3, 4, and 5. Before and after stress, samples were analyzed according to Methods 6, 7, and 8. The results are shown in Tables 2, 3, and 4.

[0424] Table 2 - Summary of SE HPLC results of samples before and after stress

[0425]

[0426] Table 3 - Summary of uniformity results by dynamic light scattering before and after stress

[0427]

[0428] Table 4 - Summary of results for acid-base distribution of samples before and after stress

[0429]

[0430] **Absolute change is calculated by:

[0431] Δ=|acid fraction content before stress - acid fraction content after stress|+|alkaline fraction content before stress - alkaline fraction content after stress|+|main fraction content before stress - main fraction content after stress|

[0432] According to the results of SE HPLC, the sample supplemented with sorbitol had the smallest increase in aggregates after thermal stress and shaking test. The formulation containing trehalose dihydrate as osmotic agent showed the largest change in acid-base profile and decreased monomer content after thermal stress; further development is not recommended.

[0433] According to the results of dynamic light scattering and according to the values ​​of the aggregation point, all samples show high thermal stability. It can also be observed that the addition of an osmotic agent increases the thermal stability of the protein in solution (the first sample without an osmotic agent shows the lowest aggregation point value in the group). When comparing the hydrodynamic radii of the compositions containing sorbitol, there is a greater tendency for the protein globules to compact.

[0434] The best stabilizers were selected using the bioinformatics prediction method of PTMs described in Method 10. According to the results of primary amino acid sequence analysis, protein degradation could potentially occur via the deamidation pathway of the side groups of some amino acids (eg, asparagine and glutamine) and methionine oxidation. Figure 1The number of potential chemical degradation sites for the light and heavy chains is shown.

[0435] The light chain may contain two isomerization sites and one deamidation site. The heavy chain contains one hydrolysis and glycosylation site, three isomerization sites, six deamidation sites, and seven oxidation sites.

[0436] Based on the results of primary protein sequence analysis of divozilimab, protein degradation could potentially occur via the deamination pathway of the side groups of some amino acids (e.g., asparagine and glutamine) and methionine oxidation.

[0437] It is suggested that L-methionine acts as a stabilizer to prevent oxidation processes. This excipient added to the pharmaceutical composition acts as a stabilizer, absorber of dissolved oxygen and absorber of free radicals. It competes with oxidation-sensitive areas within the molecule for oxidants.

[0438] In order to select stabilizers in pharmaceutical compositions, different concentrations of L-methionine and polysorbate 80 were screened.

[0439] The excipient formulations of the test pharmaceutical compositions are shown in Table 5.

[0440] Table 5 - Test drug compositions

[0441]

[0442]

[0443] Samples containing 25 mg / ml divozilimab were studied. Samples with various excipient formulations were prepared by diafiltration according to Method 2. To study the effect of various excipients, samples in the test formulations were stored at 37°C, 25°C and (2-8)°C for 5 weeks, 6 months and 24 months according to Method 11. Before and after stress, samples were analyzed according to Methods 7 and 9. The results are shown in Tables 6, 7 and 8.

[0444] Table 6 - Summary of stability results of samples stored at +37°C for 5 weeks

[0445]

[0446] Table 7 - Summary of stability results of samples stored at +25°C for 6 months

[0447]

[0448] Table 8 - Summary of stability results of samples stored at +(2-8)°C for 24 months

[0449]

[0450] The monomer content determined by SE HPLC and the acidic fraction determined by IE HPLC are shown for all temperature regimes. Figure 2 , 3 , 4, 5, 6 and 7.

[0451] The product of Formulation 2 had a smaller decrease in monomer content as determined by SE HPLC and a smaller increase in acidic fraction during storage at all temperatures.

[0452] Example 2.

[0453] According to Method 11, formulations of selected excipients were stored at a temperature regimen of 25°C, 37°C, and (2-8)°C for 5 weeks, 6 months, and 24 months at divozilimab concentrations of 25 mg / ml and 50 mg / ml, respectively.

[0454]

[0455] Stability results were obtained according to Methods 1, 7, 9, 12 and are shown in Tables 9, 10 and 11 for divozilimab concentrations of 25 mg / ml and 50 mg / ml.

[0456] Table 9 - Summary of stability results for samples stored at +37°C for 5 weeks.

[0457]

[0458]

[0459] Table 10 - Summary of stability results for samples stored at +25°C for 6 months.

[0460]

[0461] Table 11 - Summary of stability results for samples stored at +(2-8)°C for 24 months.

[0462]

[0463]

[0464] *The change is calculated by the following formula:

[0465] Δ = (fraction content at the control point - fraction content at the input control point)

[0466] **Absolute change is calculated by:

[0467] Δ=|acid fraction content before stress - acid fraction content after stress|+|alkaline fraction content before stress - alkaline fraction content after stress|+|main fraction content before stress - main fraction content after stress|

[0468] Stability data for samples with 25 mg / ml and 50 mg / ml divozilimab concentrations were comparable.

[0469] Example 3. Use of divozilimab for the treatment of patients with multiple sclerosis with exacerbations, a Phase II trial of BCD-132-2, an efficacy and safety study.

[0470] Experimental design

[0471] Study number BCD-132-2 is designed to be an international, multicenter, randomized, double-blind, double-masked, placebo-controlled trial using an active comparator (teriflunomide).

[0472] Trial No. BCD-132-2 evaluates the efficacy of divozilimab (BCD-132) in the treatment of patients with multiple sclerosis with exacerbations by brain MRI indicators and by clinical indicators related to exacerbations, and the disability level after administration of divozilimab at doses of 125 and 500 mg (compared with teriflunomide and placebo). The study includes patients diagnosed with multiple sclerosis (according to the 2017 McDonald criteria for the diagnosis of multiple sclerosis), disease course type-multiple sclerosis with exacerbations, and disability level no higher than 5.5 on the EDSS scale.

[0473] After stratification, patients were randomly assigned to the following trial groups: 125 mg divozilimab group, 500 mg divozilimab group, teriflunomide group and placebo group, with a ratio of 2:2:2:1, respectively. A total of 319 subjects were recruited in the screening phase of the trial, and 271 participants were randomized into the trial. During the main study, 24 weeks of treatment (1 treatment cycle), patients in the trial product group received the corresponding dose of divozilimab via intravenous infusion (one group had a dose of 125 mg and the other group had a dose of 500 mg). In addition, patients received the first designated dose (125 or 500 mg) via two intravenous infusions at an interval of 14 days during the first cycle (i.e., on day 1, patients received an intravenous infusion of divozilimab at a dose of 62.5 mg or 250 mg, depending on the assigned group; on day 15, the same dose was infused again). In addition, all patients randomly assigned to one of the two trial product groups received oral placebo in tablet form once a day to maintain the double-blind design. Patients in the comparator group received daily oral teriflunomide at a dose of 14 mg once daily. In addition, to maintain the double-blind design, patients in the comparator group received intravenous placebo infusions at a frequency and dose schedule similar to that of the divozilimab group in the trial. Patients in the placebo group received intravenous placebo infusions at a frequency and dose schedule similar to that of the divozilimab group in the trial (i.e., on days 1 and 15); they also received oral placebo in tablet form once daily.

[0474] Data from Milestone 1 of the trial (24 weeks of therapy) were obtained and analyzed.

[0475] Summary of clinical efficacy results from Trial No. BCD-BCD-132-2.

[0476] The results demonstrated that both trial divozilimab doses (125 mg and 500 mg) were superior to placebo and non-inferior to the comparator teriflunomide in terms of the primary endpoint, which reflects the activity of the demyelinating process in the brain of patients with MS with exacerbations and is the most reliable value for assessing therapy response during the analysis period (6 months) according to the EMA guideline on clinical investigations of medicinal products for the treatment of multiple sclerosis - "Total T1 Gd+ lesions detected by brain MRI (per scan) after 24 weeks of blinded treatment with divozilimab / teriflunomide / placebo".

[0477] The trial was based on two hypotheses: the hypothesis that the test product is superior to placebo and the hypothesis of non-inferiority between the test product BCD-132 and the comparator teriflunomide. These hypotheses were verified with respect to the efficacy endpoint, "total T1 Gd+ lesions (per scan) detected by brain MRI after 24 weeks of blinded treatment with BCD-132 / teriflunomide / placebo". The one-sided 97.5% confidence interval for the difference in the arithmetic mean of total T1 Gd+ lesions per scan between the BCD-132 (125 mg) group and the placebo group was (-Inf; -0.6075], and between the BCD-132 (500 mg) group and the placebo group was (-Inf; -0.5974]. The results in the BCD-132 group of the trial indicated that the upper limits of the confidence intervals in both groups did not exceed the pre-set superiority limit of 0. The total T1 per scan between the BCD-132 (125 mg) group and the teriflunomide comparator was 0. The two-sided 95% confidence interval for the difference in the arithmetic means of Gd+ lesions was [-0,6657; -0,1980], and [-0,6525; -0,1897] between the BCD-132 (500 mg) group and the teriflunomide group. The results in the BCD-132 group of the trial indicated that the upper limits of the confidence intervals in both groups did not exceed the pre-specified non-inferiority limit of 0,9975 (δ = 0,9975).

[0478] When comparing the arithmetic mean of total T1 Gd+ lesions per scan between the BCD-132 (125 mg) and teriflunomide groups and between the BCD-132 (500 mg) and teriflunomide groups, we observed statistically significant differences (p = 0.0004 for the comparison of the BCD-132 (125 mg) group and teriflunomide, p = 0.0005 for the comparison of the BCD-132 (500 mg) group and teriflunomide, two-sided Student-Welch t-test). Since the calculated upper limits of the confidence intervals for both BCD-132 groups did not exceed the pre-specified non-inferiority efficacy limit of 0.9975 (δ = 0.9975), the assumption of non-inferiority of the teriflunomide comparator in the primary endpoint test group has been demonstrated.

[0479] This indicates the efficacy of the therapy and achievement of the primary goal of the trial.

[0480] Dynamic estimation of the mean value of total accumulated gadolinium (Gd+) damage in T1 mode (there are two main types of MRI images: T1-weighted images and T2-weighted images). T1-weighted images register the rate at which atoms return from lateral positions to vertical positions. T2-weighted images (retention time of atoms in lateral positions after a short vertical pulse) show a reduction in damage in the divozilimab and teriflunomide groups: in the teriflunomide group, the dynamics were not statistically significant; in the divozilimab group, a statistically significant reduction relative to screening was indicated already at week 9.

[0481] Evaluation of the composite CUA (cumulative number of new T1 contrast-enhancing MRI lesions and cases of new T2 lesion patterns or T2 enlarging lesions, avoiding double counting) using per-scan analysis showed that the mean of the estimated period was statistically significantly lower in the divozilimab group compared with the teriflunomide and placebo groups (p<0.05). Dynamic observation of CUA showed a significant decrease in the mean of the index in both divozilimab groups (p<0.05; nonparametric Wilcoxon test with Benjamini-Yekutieli adjustment). There were no significant dynamics in the teriflunomide and placebo groups.

[0482] In patients without lesions, a comparable pattern in the form of reduced activity of demyelinating processes was also observed during the evaluation period. Most patients in the divozilimab group had no T1 contrast-enhancing lesions after 24 weeks of treatment (94.44% and 93.06%). These values ​​were statistically significantly higher than those in the teriflunomide and placebo groups (68.06 and 56.36%) (p<0.05).

[0483] Evaluation of other endpoints represented by MRI metrics showed statistically significant reductions in the volume of low-intensity lesions and the number of new / enlarging T2WI / FLAIR lesions over a 24-week period, including when compared between patients with new or enlarging lesions in the divozilimab group. The median annualized exacerbation frequency (ARR) in the trial group calculated using Poisson regression was lower in the teriflunomide and divozilimab (125 and 500) groups. The ARR index was statistically significantly lower in the divozilimab 500 mg group.

[0484] The EDSS (Expanded Disability Status Scale) scores showed a positive dynamics in the patients' condition, revealing a statistically significant reduction in the scores after 1 cycle of treatment in the divozilimab 500 mg group. The total proportion of patients who continued to progress after 24 weeks of treatment was 4.17% in the teriflunomide and BCD-132 (125 mg) groups, 1.39% in the BCD-132 (500 mg) group, and 7.27% in the placebo group (p = 0.3999; Fisher's exact test). There was no significant negative dynamics in the groups when evaluating neurological deficits and cognitive function using a number of other scales and questionnaires (Timed 7.62 m / 25 ft Walking Test, 9-Hole Pillar Test (9-NHT), Symbol Digit Form Test (SDMT)), as well as when evaluating the patients' quality of life (SF-36, EQ-5D, MSQOL-54).

[0485] The primary efficacy results after 24 weeks of therapy in the BCD-132-2 trial are shown in Table 12.

[0486] Table 12 - Key efficacy parameters after 24 weeks of treatment in the BCD-132-2 trial

[0487]

[0488]

[0489]

[0490] The proportion of patients with unconfirmed exacerbations is shown in Table 13. In the BCD-132 group, by the 24th week of therapy, the proportions were 93.06% and 97.22% of patients (corresponding to 125 mg and 500 mg, respectively). Before glucocorticoid (GC) therapy, all patients with exacerbations underwent brain MRI. If the MRI results revealed contrast-enhanced T1 lesions and new and / or enlarged T2-WI and T2-FLAIR lesions, such exacerbations were considered to be MRI-confirmed exacerbations. By the 24th week of therapy, there were no such exacerbations in the divozilimab group, while in the teriflunomide and placebo groups, the proportions were 5.56% and 7.27%, respectively. The proportion of patients with exacerbations confirmed by AINC (Assessment Independent Neurological Committee) was 6.94% and 1.39% in the BCD-132 group (corresponding to 125 mg and 500 mg, respectively), compared to 11.11% and 20.00% in the TFL and PBO groups, respectively.

[0491] Table 13 - Primary efficacy parameters associated with exacerbations after 24 weeks of therapy in the BCD-132-2 trial

[0492]

[0493] Dynamic evaluation of individual key parameters of brain MRI and clinical indicators demonstrated a positive treatment effect: reduced activity of demyelinating processes in the brain and fewer exacerbations in patients. A slightly more pronounced effect was observed in the group receiving the higher trial divozilimab dose compared with the lower dose group, with comparable safety parameters.

[0494] The data support the efficacy of divozilimab in the treatment of patients with MS with exacerbations.

[0495] Summary of clinical safety results from trial number BCD-132-2.

[0496] All trial groups demonstrated an acceptable safety profile throughout the evaluation period. The therapy was satisfactorily tolerated in the target population of patients with multiple sclerosis with exacerbations. The analysis using safety endpoints did not reveal any statistically significant differences. According to the investigators, during the analysis period, the AEs related to the trial intravenous medicinal product in organs and systems were mainly (more than 5% of patients) respiratory tract infections, infusion reactions, deviations in laboratory and instrumental data (neutropenia, lymphopenia, leukopenia). In particular, we also observed infectious conditions, such as: infections caused by herpes viruses and oral herpes, infections caused by Candida fungi and enterovirus infections, urinary tract infections and cystitis, pneumonia.

[0497] Deviations in other organs and systems were observed in less than 5% of patients. The registered AEs were mainly of mild and moderate (grade 1-2). According to the investigators, there were no statistically significant differences between the groups in any deviations related to the trial intravenous drug. According to the investigators, AEs of grade 3-4 according to CTCAE v.5.0 related to intravenous drugs were represented by deviations in laboratory and instrumental data for hepato / biliary disorders and vascular disorders.

[0498] in conclusion

[0499] The study was conducted on a representative cohort, which was balanced by key demographic parameters and disease characteristics, and which approximates the target population of patients with multiple sclerosis.

[0500] The resulting efficacy and safety data justified the use of 125 and 500 mg doses of divozilimab.

[0501] The multicenter, open-label Phase II trial No. BCD-132-2 in a population of patients with multiple sclerosis demonstrated a significant therapeutic effect with divozilimab. Considering the efficacy indicators obtained in this trial with a satisfactory safety profile, the use of divozilimab provides a risk-benefit balance in the given patient population.

Claims

1. A pharmaceutical composition of an anti-CD20 antibody, comprising: (i) anti-CD20 antibodies; (ii) acetate buffer; (iii) sorbitol; (iv) methionine; (v) Water for injection.

2. The pharmaceutical composition according to claim 1, wherein the anti-CD20 antibody is present at a concentration of 1.5-60.0 mg / ml.

3. The pharmaceutical composition according to claim 1, wherein the anti-CD20 antibody is present at a concentration of 5.0-50.0 mg / ml.

4. The pharmaceutical composition according to claim 1, wherein the anti-CD20 antibody is present at a concentration of 10.0-40.0 mg / ml.

5. The pharmaceutical composition according to claim 1, wherein the anti-CD20 antibody is present at a concentration of 20.0-30.0 mg / ml.

6. The pharmaceutical composition of claim 1, wherein the anti-CD20 antibody is present at a concentration of 25.0 mg / ml.

7. The pharmaceutical composition of claim 1, wherein the anti-CD20 antibody is divozilimab.

8. The pharmaceutical composition according to claim 1, wherein the acetate buffer is a mixture of sodium acetate and acetic acid.

9. The pharmaceutical composition of claim 8, wherein sodium acetate is present in a concentration of 0.5-3.0 mg / ml.

10. The pharmaceutical composition of claim 8, wherein sodium acetate is present in a concentration of 1.0-2.0 mg / ml.

11. The pharmaceutical composition of claim 8, wherein sodium acetate is present in a concentration of 1.5-2.0 mg / ml.

12. The pharmaceutical composition of claim 8, wherein sodium acetate is present at a concentration of 1.74 mg / ml.

13. The pharmaceutical composition according to claim 8, wherein the sodium acetate is sodium acetate trihydrate.

14. The pharmaceutical composition according to claim 8, wherein acetic acid is added to pH 5.0-6.

0.

15. The pharmaceutical composition according to claim 8, wherein acetic acid is added to pH 5.4-5.

6.

16. The pharmaceutical composition according to claim 8, wherein acetic acid is added to pH 5.

5.

17. The pharmaceutical composition according to claim 8, wherein the acetic acid is glacial acetic acid.

18. The pharmaceutical composition of claim 1, wherein sorbitol is present in a concentration of 40.0-60.0 mg / ml.

19. The pharmaceutical composition of claim 1, wherein sorbitol is present in a concentration of 45.0-55.0 mg / ml.

20. The pharmaceutical composition of claim 1, wherein sorbitol is present at a concentration of 50.0 mg / ml.

21. The pharmaceutical composition of claim 1, wherein methionine is present at a concentration of 0.05-0.30 mg / ml.

22. The pharmaceutical composition of claim 1, wherein methionine is present at a concentration of 0.10-0.20 mg / ml.

23. The pharmaceutical composition of claim 1, wherein methionine is present at a concentration of 0.15 mg / ml.

24. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibodies; (ii) an acetate buffer which is a mixture of: Sodium acetate 0.5-3.0 mg / ml and acetic acid to pH 5.0-6.0; (iii) sorbitol 40.0-60.0 mg / ml; (iv) methionine 0.05-0.30 mg / ml; (v) water for injection up to 1 ml.

25. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibodies; (ii) an acetate buffer which is a mixture of: Sodium acetate 1.0-2.0 mg / ml and acetic acid to pH 5.4-5.6; (iii) sorbitol 45.0-55.0 mg / ml; (iv) methionine 0.10-0.20 mg / ml; (v) water for injection up to 1 ml.

26. The pharmaceutical composition according to claim 1, comprising: (ii) anti-CD20 antibodies; (iii) acetate buffer which is a mixture of: Sodium acetate 1.5-2.0 mg / ml and acetic acid to pH 5.4-5.6; (vi) sorbitol 45.0-55.0 mg / ml; (vii) methionine 0.10-0.20 mg / ml; (viii) water for injection to 1 ml.

27. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibodies; (ii) an acetate buffer which is a mixture of: Sodium acetate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) Water for injection to 1 ml.

28. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibodies; (ii) an acetate buffer which is a mixture of: Sodium acetate trihydrate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) water for injection up to 1 ml.

29. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibody 25.0 mg / ml; (ii) an acetate buffer which is a mixture of: Sodium acetate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) water for injection up to 1 ml.

30. The pharmaceutical composition according to claim 1, comprising: (i) divozilimab 25.0 mg / ml; (ii) an acetate buffer which is a mixture of: Sodium acetate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) water for injection up to 1 ml.

31. The pharmaceutical composition according to claim 1, comprising: (i) anti-CD20 antibody 25.0 mg / ml; (ii) an acetate buffer which is a mixture of: Sodium acetate trihydrate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) Water for injection to 1 ml.

32. The pharmaceutical composition according to claim 1, comprising: (i) divozilimab 25.0 mg / ml; (ii) an acetate buffer which is a mixture of: Sodium acetate trihydrate 1.74 mg / ml and acetic acid to pH 5.5; (iii) sorbitol 50.0 mg / ml; (iv) methionine 0.15 mg / ml; (v) Water for injection to 1 ml.

33. Use of a pharmaceutical composition according to any one of claims 1-32 for treating a CD20-mediated disease or disorder in a subject in need thereof.

34. The use according to claim 33, wherein the pharmaceutical composition is administered at a dose of 125 mg of anti-CD20 antibody.

35. The use according to claim 33, wherein the pharmaceutical composition is administered at a dose of 500 mg of anti-CD20 antibody.

36. The use according to claim 33, wherein the pharmaceutical composition is administered every 24 weeks.

37. The use according to claim 33, wherein the pharmaceutical composition is administered every 24 weeks at a dose of 125 mg of the anti-CD20 antibody.

38. The use according to claim 33, wherein the pharmaceutical composition is administered at a dose of 500 mg of the anti-CD20 antibody every 24 weeks.

39. The use according to claim 33, comprising administering the pharmaceutical composition for at least 5 cycles.

40. The use according to claim 33, comprising administering the pharmaceutical composition for at least 5 cycles, wherein cycle 1 comprises a first administration of the pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody and a second administration of the pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody 2 weeks after the first administration; and subsequent cycles comprise administering the pharmaceutical composition at a dose of 125 mg of an anti-CD20 antibody every 24 weeks after the first administration.

41. The use according to claim 33, comprising administering the pharmaceutical composition for at least 5 cycles, wherein cycle 1 comprises a first administration of the pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody and a second administration of the pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody 2 weeks after the first administration; and subsequent cycles comprise administering the pharmaceutical composition at a dose of 500 mg of an anti-CD20 antibody every 24 weeks after the first administration.

42. The use according to claim 33, wherein the anti-CD20 antibody is divozilimab.

43. The use according to claim 33, wherein the CD20-mediated disease or condition is selected from: (i) a neoplastic disease or condition, or (ii) Autoimmune diseases or disorders.

44. Use according to claim 43, wherein the neoplastic disease or condition is selected from: B-cell lymphoma, leukemia.

45. The use according to claim 44, wherein the B cell lymphoma is non-Hodgkin's lymphoma, Hodgkin's lymphoma.

46. ​​The use according to claim 44, wherein the leukemia is selected from the group consisting of chronic lymphocytic leukemia and small lymphocytic lymphoma.

47. The use of claim 43, wherein the autoimmune disease or disorder is selected from the group consisting of multiple sclerosis, axonal neuropathy, ANCA-associated vasculitis, IgG4-associated disease, amyloidosis, axial spondyloarthritis, antiphospholipid syndrome, Takayasu's arteritis, autoimmune aplastic anemia, autoimmune inner ear disease, autoimmune hemolytic anemia, autoimmune hyperlipidemia, autoimmune familial dysautonomia, autoimmune urticaria, autoimmune neuromyotonia (Isaacs syndrome), autoimmune retinopathy, autoimmune thrombocytopenic purpura, warm antibody hemolytic anemia, autoimmune testicular disease, autoimmune angioedema, autoimmune hepatitis, autoimmune diabetes, Autoimmune immunodeficiency, autoimmune pericarditis, autoimmune thyroiditis, autoimmune progesterone dermatitis, Addison's disease, Behcet's disease, Graves' disease (diffuse toxic goiter), neuromyelitis optica spectrum (Devic's disease), Castleman's disease, Crohn's disease, Ormond's disease (retroperitoneal fibrosis), Raynaud's disease, Churg-Strauss syndrome, bullous pemphigoid, epidermolysis bullosa, vasculitis, renal vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, cystic dermatosis, giant cell arteritis, vitiligo, lupus nephritis, inflammatory aortic aneurysm, inflammatory bowel disease, congenital heart block, pemphigus vulgaris, dermatitis herpetiformis, pemphigoid gestationis, hypogammaglobulinemia, glomerulonephritis, granulomatosis with polyangiitis (Wegener syndrome), dermatomyositis, dilated cardiomyopathy, demyelinating neuropathy, discoid lupus erythematosus, idiopathic recurrent gross hematuria (IgA nephropathy), idiopathic inflammatory myopathy, idiopathic inflammatory pseudotumor, idiopathic thrombocytopenic purpura, idiopathic hypocomplementemic tubulointerstitial nephritis, idiopathic pulmonary fibrosis, diabetes mellitus, pure red cell aplasia, Barlow's concentric sclerosis, lichen planus, lichen sclerosus, leukocytoclastic vasculitis, limited scleroderma (CREST syndrome), limbic encephalitis, linear IgA bullous dermatosis, pemphigus foliaceus, Waldenstrom's macroglobulinemia, mediastinal fibrosis, myasthenic crisis, Lang-Evans myasthenic syndrome, myasthenia gravis, microscopic polyangiitis, myositis, coxsackie myocarditis inflammation, multiple sclerosis, multifocal motor neuropathy, multifocal fibrosclerosis, optic neuritis, undifferentiated connective tissue disease uveitis, neutropenia, ulcerative colitis, anti-tubular / membrane antibody-associated nephritis, strabismus clonus, acute motor axonal neuropathy, acute hemorrhagic leukoencephalitis, acute disseminated encephalomyelitis, relapsing rheumatic disease, paraneoplastic cerebellar degeneration, paraneoplastic syndrome, paraproteinemic polyneuropathy, paroxysmal nocturnal hemoglobinuria, ocular pemphigus, mucous membrane pemphigoid, primary biliary cirrhosis, primary sclerosing cholangitis, periaortitis, perianal arteritis, perivenous encephalomyelitis, peripheral neuropathy, peripheral uveitis, pernicious anemia, subacute bacterial endocarditis, polymyositis,Polyneuropathy with organomegaly (POEMS syndrome), transverse myelitis, post-myocardial infarction syndrome, post-traumatic pericarditis, psoriasis, psoriatic arthritis, pemphigus, Henoch-Schönlein purpura, anti-HLA antibody-mediated transplant rejection, reactive arthritis, rheumatic fever, polymyalgia rheumatoid, rheumatoid arthritis, reflex sympathetic dystrophy, refractory epilepsy, relapsing polychondritis, sarcoidosis, acute febrile polyneuritis, Kawasaki syndrome, Cogan syndrome, Mikulic syndrome, stiff-man syndrome, Sjögren syndrome, Evans syndrome, systemic lupus erythematosus, systemic scleroderma, localized scleroderma, Riedel's thyroiditis, Hashimoto's thyroiditis, Thyroid-related eye disease, thyroid storm, thrombotic thrombocytopenic purpura, polyarteritis nodosa, fetal and neonatal alloimmune thrombocytopenia, fibrosing alveolitis, B-cell non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia, cold blood cell agglutinin disease, Sydenham's chorea (small chorea or rheumatic chorea), chronic Lyme disease, chronic inflammatory demyelinating polyneuropathy, chronic relapsing multifocal osteomyelitis, chronic sclerosing sialadenitis (Tutnot tumor), celiac disease, endometriosis, encephalitis, autoantibody-related encephalopathy, Hashimoto's encephalitis, eosinophilic fasciitis, essential mixed cryoglobulinemia, juvenile diabetes, juvenile rheumatoid arthritis.

48. The use according to claim 47, wherein the multiple sclerosis is remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, multiple sclerosis with exacerbations, highly active multiple sclerosis, aggressive multiple sclerosis, malignant multiple sclerosis (Marburg variant), myelin-destroying diffuse Schilder sclerosis, myelocortical multiple sclerosis, clinically isolated syndrome.

49. A method of treating a CD20-mediated disease or disorder comprising administering a pharmaceutical composition according to any one of claims 1-30 in a therapeutically effective amount to a subject in need thereof.

50. The method of treating a disease or condition according to claim 49, wherein the pharmaceutical composition is administered at a dose of 125 mg of anti-CD20 antibody.

51. The method of treating a disease or condition according to claim 49, wherein the pharmaceutical composition is administered at a dose of 500 mg of anti-CD20 antibody.

52. The method of treating a disease or condition according to claim 49, wherein the pharmaceutical composition is administered every 24 weeks.

53. The method of treating a disease or condition according to claim 49, wherein the pharmaceutical composition is administered every 24 weeks at a dose of 125 mg of the anti-CD20 antibody.

54. The method of treating a disease or condition according to claim 49, wherein the pharmaceutical composition is administered every 24 weeks at a dose of 500 mg of the anti-CD20 antibody.

55. The method of treating a disease or condition according to claim 49, comprising administering at least 5 cycles of the pharmaceutical composition.

56. The method of treating a disease or condition according to claim 49, comprising at least 5 cycles, wherein cycle 1 comprises a first administration of the pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody and a second administration of the pharmaceutical composition at a dose of 62.5 mg of an anti-CD20 antibody 2 weeks after the first administration; subsequent cycles comprise administration of the pharmaceutical composition at a dose of 125 mg of an anti-CD20 antibody every 24 weeks after the first administration.

57. The method for treating a disease or condition according to claim 49, comprising at least 5 cycles, wherein cycle 1 comprises a first administration of the pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody and a second administration of the pharmaceutical composition at a dose of 250 mg of an anti-CD20 antibody 2 weeks after the first administration; and subsequent cycles comprise administration of the pharmaceutical composition at a dose of 500 mg of an anti-CD20 antibody every 24 weeks after the first administration.

58. The method of treating a disease or condition of claim 49, wherein the anti-CD20 antibody is divozilimab.

59. The method of treating a disease or condition according to claim 49, wherein the CD20-mediated disease or condition is selected from: (i) a neoplastic disease or condition, or (ii) Autoimmune diseases or disorders.

60. The method of treating a disease or condition according to claim 59, wherein the neoplastic disease or condition is selected from the group consisting of: B cell lymphoma, leukemia.

61. The method of treating a disease or condition according to claim 60, wherein the B cell lymphoma is non-Hodgkin lymphoma, Hodgkin lymphoma.

62. The method of treating a disease or condition according to claim 60, wherein the leukemia is selected from the group consisting of: chronic lymphocytic leukemia, small lymphocytic lymphoma.

63. The method of claim 59 for treating a disease or condition, wherein the autoimmune disease or condition is selected from the group consisting of: multiple sclerosis, axonal neuropathy, ANCA-associated vasculitis, IgG4-associated disease, amyloidosis, axial spondyloarthritis, antiphospholipid syndrome, Takayasu's arteritis, autoimmune aplastic anemia, autoimmune inner ear disease, autoimmune hemolytic anemia, autoimmune hyperlipidemia, autoimmune familial dysautonomia, autoimmune urticaria, autoimmune neuromyotonia (Isaacs syndrome), autoimmune retinopathy, autoimmune thrombocytopenic purpura, warm antibody hemolytic anemia, autoimmune testicular disease, autoimmune angioedema, autoimmune hepatitis, autoimmune Immune diabetes, autoimmune immune deficiency, autoimmune pericarditis, autoimmune thyroiditis, autoimmune progesterone dermatitis, Addison's disease, Behcet's disease, Graves' disease (diffuse toxic goiter), neuromyelitis optica spectrum (Devic's disease), Castleman's disease, Crohn's disease, Ormond's disease (retroperitoneal fibrosis), Raynaud's disease, Churg-Strauss syndrome, bullous pemphigoid, epidermolysis bullosa, vasculitis, renal vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, cystic dermatosis, giant cell arteritis, vitiligo, lupus nephritis, inflammatory aortic aneurysm, inflammatory bowel disease, congenital heart block, pemphigus vulgaris, dermatitis herpetiformis, pemphigoid gestationis, hypogammaglobulinemia, glomerulonephritis, granulomatous disease with Polyangiitis (Wegener syndrome), dermatomyositis, dilated cardiomyopathy, demyelinating neuropathy, discoid lupus erythematosus, idiopathic recurrent gross hematuria (IgA nephropathy), idiopathic inflammatory myopathy, idiopathic inflammatory pseudotumor, idiopathic thrombocytopenic purpura, idiopathic hypocomplementemic tubulointerstitial nephritis, idiopathic pulmonary fibrosis, diabetes mellitus, pure red cell aplasia, Barlow's concentric sclerosis, lichen planus, lichen sclerosus, leukocytoclastic vasculitis, limited scleroderma (CREST syndrome), limbic encephalitis, linear IgA bullous dermatosis, pemphigus foliaceus, Waldenstrom's macroglobulinemia, mediastinal fibrosis, myasthenic crisis, Langer-Evans syndrome, myasthenia gravis, microscopic polyangiitis, myositis, coxsackie Myocarditis, multiple sclerosis, multifocal motor neuropathy, multifocal fibrosclerosis, optic neuritis, undifferentiated connective tissue disease uveitis, neutropenia, ulcerative colitis, anti-tubular / membrane antibody-associated nephritis, strabismus opsoclonus, acute motor axonal neuropathy, acute hemorrhagic leukoencephalitis, acute disseminated encephalomyelitis, relapsing rheumatic disease, paraneoplastic cerebellar degeneration, paraneoplastic syndrome, paraproteinemic polyneuropathy, paroxysmal nocturnal hemoglobinuria, ocular pemphigus, mucous membrane pemphigoid, primary biliary cirrhosis, primary sclerosing cholangitis, periaortitis, perianal arteritis, perivenous encephalomyelitis, peripheral neuropathy, peripheral uveitis, pernicious anemia, subacute bacterial endocarditis, polymyositis,Polyneuropathy with organomegaly (POEMS syndrome), transverse myelitis, post-myocardial infarction syndrome, post-traumatic pericarditis, psoriasis, psoriatic arthritis, pemphigus, Henoch-Schönlein purpura, anti-HLA antibody-mediated transplant rejection, reactive arthritis, rheumatic fever, polymyalgia rheumatoid, rheumatoid arthritis, reflex sympathetic dystrophy, refractory epilepsy, relapsing polychondritis, sarcoidosis, acute febrile polyneuritis, Kawasaki syndrome, Cogan syndrome, Mikulic syndrome, stiff-man syndrome, Sjögren syndrome, Evans syndrome, systemic lupus erythematosus, systemic scleroderma, localized scleroderma, Riedel's thyroiditis, Hashimoto's thyroiditis, Thyroid-related eye disease, thyroid storm, thrombotic thrombocytopenic purpura, polyarteritis nodosa, fetal and neonatal alloimmune thrombocytopenia, fibrosing alveolitis, B-cell non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia, cold blood cell agglutinin disease, Sydenham's chorea (small chorea or rheumatic chorea), chronic Lyme disease, chronic inflammatory demyelinating polyneuropathy, chronic relapsing multifocal osteomyelitis, chronic sclerosing sialadenitis (Tutnot tumor), celiac disease, endometriosis, encephalitis, autoantibody-related encephalopathy, Hashimoto's encephalitis, eosinophilic fasciitis, essential mixed cryoglobulinemia, juvenile diabetes, juvenile rheumatoid arthritis.

64. The method of treating a disease or disorder according to claim 63, wherein the multiple sclerosis is remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, multiple sclerosis with exacerbations, highly active multiple sclerosis, aggressive multiple sclerosis, malignant multiple sclerosis (Marburg variant), myelin-destroying diffuse Schilder sclerosis, myelocortical multiple sclerosis, clinically isolated syndrome.

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