Use of Anti-CD20 antibody or antigen-binding fragment thereof in treatment of diseases

Abstract

The present invention relates to use of an anti-CD20 antibody or an antigen-binding fragment thereof in the preparation of a drug for treating diseases associated with CD20 expression.

Description

Application of anti-CD20 antibodies or their antigen-binding fragments in the treatment of diseases Technical Field

[0001] This invention belongs to the field of biopharmaceuticals, specifically relating to a method for treating diseases, and the use of anti-CD20 antibodies or their antigen-binding fragments in treating diseases. Background Technology

[0002] CD20 is a non-glycosylated phosphoprotein specifically labeled on the surface of human lymphocyte subsets (B cells). It consists of 297 amino acids, has a molecular weight of 33-37 kDa, and is expressed on the surface of over 95% of B cells. CD20 is present in both normal and malignant B cells, particularly in over 90% of B-cell non-Hodgkin's lymphomas. The CD20 molecule has four transmembrane regions, with the N-terminus and C-terminus located on the inner side of the plasma membrane. Between the third and fourth transmembrane regions is a loop region consisting of 43 amino acid residues, forming the major epitope. The CD20 antigen molecule is relatively exposed and easily accessible. When CD20 molecules approach each other under the action of antibodies, the polymers formed through cross-linking or even hypercross-linking act as calcium ion channels, allowing extracellular calcium ions to flow into the cell. Furthermore, Src family tyrosine protein kinases are mutually activated due to proximity. This activation of signaling pathways and endogenous calcium stores leads to an increase in intracellular calcium ion concentration, thereby affecting cell cycle progression, regulating cell proliferation and differentiation, and even inducing apoptosis.

[0003] Although the actual role of CD20 in promoting B cell proliferation and differentiation is still uncertain, CD20 provides an important target for antibody-mediated therapy that can be used to control B cells involved in cancer and autoimmune diseases. Summary of the Invention

[0004] Based on this, the present invention provides the application of anti-CD20 antibody or its antigen-binding fragment in the treatment of diseases.

[0005] On one hand, the present invention provides a method for treating a disease, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region (VL) and a heavy chain variable region (VH), said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0006] The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 710 mg to approximately 790 mg, for example, approximately 750 mg.

[0007] In some implementations, the patient is a person.

[0008] In some implementations, the patient has a disease characterized by CD20 positivity.

[0009] In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered by injection, such as intravenous injection, subcutaneous injection, or intraperitoneal injection; or, the administration method is intravenous infusion.

[0010] In some implementations, the disease is one associated with CD20 expression, such as tumorigenic diseases and immune diseases.

[0011] In some embodiments, the oncogenic disease includes B-cell lymphoma, which includes precursor B-cell lymphocytic leukemia / lymphoma and mature B-cell tumors, lymphoplasmacytic lymphoma, mantle cell lymphoma (MCL), low-grade, intermediate-grade and high-grade follicular lymphoma, cutaneous follicular center lymphoma, MALT-type, nodular and splenic marginal zone B-cell lymphoma, hairy cell leukemia, diffuse large B-cell lymphoma, Burkitt lymphoma, plasmacytoma, plasmacytic myeloma, post-transplant lymphoproliferative disorder, Waldenström macroglobulinemia and anaplastic large cell lymphoma (ALCL).

[0012] In some implementations, the immune diseases include psoriasis, psoriatic arthritis, dermatitis, systemic scleroderma and sclerosis, inflammatory bowel disease (IBD), segmental ileitis, ulcerative colitis, acute respiratory distress syndrome, meningitis, encephalitis, uveitis, nephrotic syndrome (e.g., glomerulonephritis), eczema, asthma, atherosclerosis, leukocyte adhesion defect, multiple sclerosis, Raynaud's syndrome, Sjögren's syndrome, juvenile diabetes mellitus, Leter's disease, Behçet's disease, immune complex nephritis, IgA nephropathy, IgM polyneuropathy, neuromyelitis optica spectrum disorders, immune-mediated thrombocytopenic purpura, hemolytic anemia, myasthenia gravis, lupus nephritis, systemic lupus erythematosus, rheumatoid arthritis (RA), atopic dermatitis, pemphigus, Graves' disease, Hashimoto's thyroiditis, Wechsler's granulomatosis, Omenn's syndrome, chronic renal failure, acute infectious mononucleosis, and chronic obstructive pulmonary disease.

[0013] In some implementations, the disease is a disease or condition that can be treated by B-cell depletion.

[0014] In some implementations, the diseases or conditions that can be treated by B-cell depletion are selected from cancer, allergic reactions, cardiovascular diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, neurological diseases, viral infections, and bacterial infections. For example, the disease may be cancer or an allergic reaction.

[0015] In some implementations, the disease is selected from pernicious anemia (Addison's disease), amyotrophic lateral sclerosis, ankylosing spondylitis, erythema multiforme, lupus nephritis, dermatomyositis, immune-mediated thrombocytopenic purpura, such as acute idiopathic thrombocytopenic purpura and chronic idiopathic thrombocytopenic purpura, Siddenham's chorea, rheumatic fever, polygonococcal syndrome, allergic purpura, post-streptococcal nephritis, erythema nodosum, aortitis, polyarteritis nodosum, pulmonary hemorrhage nephritis syndrome, thromboangiitis obliterans, primary biliary cirrhosis, Hashimoto's thyroiditis, thyrotoxicosis, chronic active hepatitis, etc. Polymyositis / dermatomyositis, polychondritis, pemphigus vulgaris, Wechsler's granulomatosis, membranous nephropathy, tabes dorsalis, polymyalgia, pernicious anemia, rapidly progressive glomerulonephritis and fibrotic alveolitis, inflammatory diseases such as inflammatory skin diseases, including psoriasis and dermatitis (e.g., atopic dermatitis); systemic scleroderma and sclerosis; diseases associated with inflammatory bowel disease (e.g., segmental ileitis and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS), dermatitis, meningitis, encephalitis, colitis, nephrotic syndrome (e.g., glomerulonephritis), uveitis, allergic diseases such as eczema and asthma, and those involving Other diseases involving T-cell infiltration and chronic inflammation; atherosclerosis; leukocyte adhesion defects; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes (e.g., type 1 diabetes or insulin-dependent diabetes); multiple sclerosis; Raynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjögren's syndrome; juvenile diabetes; and cytokine- and T-lymphocyte-mediated immune responses associated with acute and delayed-type hypersensitivity reactions, commonly found in tuberculosis, sarcoidosis, polymyositis, granulomatous diseases, and vasculitis; diseases involving leukocyte infiltration; and inflammation of the central nervous system (CNS). Sexual disorders; multiple organ injury syndrome; hemolytic anemia (including but not limited to cryo-saturated glomerulonephritis or Coulomb positive anemia); myasthenia gravis; antigen-antibody complex-mediated diseases; antiglomerular basement membrane diseases; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; bullous pemphigoid; pemphigus; autoimmune polyendocrine disorders; Leter's disease; stiff-person syndrome; Behçet's disease; giant cell arteritis; immune complex nephritis; IgA nephropathy; IgM polyneuropathy; immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia, etc.

[0016] In some implementations, the disease is neuromyelitis optica spectrum disorder (NMOSD), non-Hodgkin's lymphoma (NHL), multiple sclerosis (MS), immune thrombocytopenic purpura (ITP), rheumatoid arthritis (RA), Wechsler granulomatosis (WG), microscopic polyangiitis (MPA), lupus nephritis, systemic lupus erythematosus, or chronic lymphocytic leukemia (CLL).

[0017] Neuromyelitis optica (NMO) is an immune-mediated inflammatory demyelinating disease of the central nervous system (CNS), primarily affecting the optic nerve and spinal cord. Its main characteristics are severe optic neuritis (ON) and long segmental transverse myelitis (LETM). With further research, clinical features of NMO have also included some non-visual and non-spinal manifestations. The etiology of NMO is primarily related to aquaporin 4 antibody (AQP4-IgG). In addition, a group of limited forms of demyelinating diseases do not clinically meet the diagnostic criteria for NMO. Their pathogenesis and clinical features are similar to NMO, and some cases eventually evolve into NMO. In 2007, these diseases were collectively referred to as neuromyelitis optica spectrum disorders (NMOSD). In 2015, the International NMO Diagnostic Panel (IPND) reached a new international consensus on the diagnostic criteria for NMOSD, unifying NMO and NMOSD under the single term NMOSD.

[0018] In some implementations, NMOSD is characterized by optic neuritis (ON) and long segmental transverse myelitis (LETM). In some implementations, NMOSD is diagnosed according to the diagnostic criteria for NMOSD established by the International NMO Diagnostic Panel (IPND). In some implementations, NMOSD has experienced at least two relapses within two years. In some implementations, NMOSD has experienced at least one relapse within one year. In some implementations, NMOSD is AQP4-IgG negative. In some implementations, NMOSD is AQP4-IgG positive.

[0019] In some embodiments, NMOSD is characterized by optic neuritis (ON). In some embodiments, NMOSD is characterized by acute myelitis. In some embodiments, NMOSD is characterized by area posterior syndrome (APS). In some embodiments, NMOSD is characterized by acute brainstem syndrome or other brainstem syndromes. In some embodiments, NMOSD is characterized by symptomatic narcolepsy. In some embodiments, NMOSD is characterized by acute diencephalic syndrome. In some embodiments, NMOSD is characterized by symptomatic diencephalic syndrome. In some embodiments, NMOSD is characterized by cerebral syndrome. In some embodiments, NMOSD is characterized by episodic hiccups, nausea, and vomiting without other identifiable cause. In some embodiments, NMOSD is characterized by diencephalic lesions characteristic of NMOSD on brain MRI. In some embodiments, NMOSD is characterized by cerebral syndrome with characteristic cerebral lesions of NMOSD. In some embodiments, NMOSD is characterized by area posterior syndrome.

[0020] In some embodiments, the disease is refractory to at least one prior treatment regimen or has relapsed after at least one prior treatment regimen. In some embodiments, the disease is refractory to at least two prior treatment regimens or has relapsed after at least two prior treatment regimens. In some embodiments, the disease has been treated with NMOSD, such as glucocorticoids, gabapentin, neurological drugs, selective immunosuppressants, and / or other immunosuppressants. In some embodiments, prior treatment regimens include standard treatments for treating acute exacerbations, such as corticosteroids (e.g., methylprednisolone) and / or plasma exchange. In some embodiments, prior treatment regimens include standard treatments for preventing exacerbations, such as eculizumab (a humanized monoclonal antibody) and / or other immunosuppressants (e.g., azathioprine, rituximab, and mycophenolate mofetil). In some embodiments, prior treatment regimens include systemic steroids, antitumor and immunomodulatory drugs, and neurological drugs. In some embodiments, the systemic steroids do not include sex hormones and insulin. In some embodiments, prior treatment regimens include corticosteroids (e.g., glucocorticoids), selective immunosuppressants, gabapentin, nervous system drugs, immunosuppressants, antipyretics, antidepressants and tranquilizers, and amantadine derivatives, or combinations thereof. In some embodiments, the prior treatment regimen is glucocorticoids. In some embodiments, the disease has not been treated with non-pharmacological therapies.

[0021] In some embodiments, the neuromyelitis optica spectrum disorder (NMOSD) is refractory to at least one prior treatment regimen or has relapsed after at least one prior treatment regimen. In some embodiments, the NMOSD is refractory to at least two prior treatment regimens or has relapsed after at least two prior treatment regimens. In some embodiments, the NMOSD has been treated with NMOSD, such as glucocorticoids, gabapentin, neurological drugs, selective immunosuppressants, and / or other immunosuppressants. In some embodiments, the prior treatment regimen includes standard treatment for treating acute exacerbations, such as corticosteroids (e.g., methylprednisolone) and / or plasma exchange. In some embodiments, the prior treatment regimen includes standard treatment for preventing exacerbations, such as eculizumab (a humanized monoclonal antibody) and / or other immunosuppressants (e.g., azathioprine, rituximab, and mycophenolate mofetil). In some embodiments, the prior treatment regimen includes systemic steroids, antitumor and immunomodulatory drugs, and neurological drugs. In some embodiments, the systemic steroids do not include sex hormones and insulin. In some embodiments, prior treatment regimens include corticosteroids (e.g., glucocorticoids), selective immunosuppressants, gabapentin, nervous system drugs, immunosuppressants, antipyretics, antidepressants and tranquilizers, and amantadine derivatives, or combinations thereof. In some embodiments, the prior treatment regimen is glucocorticoids. In some embodiments, the neuromyelitis optica spectrum disorder has not been treated with non-pharmacological therapies.

[0022] In some embodiments, the present invention provides a method for treating AQP4-IgG negative neuromyelitis optica spectrum disorder, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0023] The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 10 mg to approximately 3000 mg.

[0024] In some embodiments, the AQP4-IgG negative neuromyelitis optica spectrum disorder is refractory to at least one prior treatment regimen or has relapsed after at least one prior treatment regimen. In some embodiments, the AQP4-IgG negative neuromyelitis optica spectrum disorder is refractory to at least two prior treatment regimens or has relapsed after at least two prior treatment regimens. In some embodiments, the AQP4-IgG negative neuromyelitis optica spectrum disorder has been treated with NMOSD, such as glucocorticoids, selective immunosuppressants, and / or other immunosuppressants. In some embodiments, the prior treatment regimen includes standard treatment for treating acute exacerbations, such as corticosteroids (e.g., methylprednisolone) and / or plasma exchange. In some embodiments, the prior treatment regimen includes standard treatment for preventing exacerbations, such as eculizumab (a humanized monoclonal antibody) and / or other immunosuppressants (e.g., azathioprine, rituximab, and mycophenolate mofetil). In some embodiments, the AQP4-IgG negative neuromyelitis optica spectrum disorder has not received non-pharmacological treatment.

[0025] In some embodiments, the present invention provides a method for treating AQP4-IgG positive neuromyelitis optica spectrum disorder, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0026] The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 10 mg to approximately 3000 mg.

[0027] In some embodiments, the AQP4-IgG positive neuromyelitis optica spectrum disorder is refractory to at least one prior treatment regimen or has relapsed after at least one prior treatment regimen. In some embodiments, the AQP4-IgG positive neuromyelitis optica spectrum disorder is refractory to at least two prior treatment regimens or has relapsed after at least two prior treatment regimens. In some embodiments, the AQP4-IgG positive neuromyelitis optica spectrum disorder has been treated with NMOSD, such as glucocorticoids, selective immunosuppressants, and / or other immunosuppressants. In some embodiments, the prior treatment regimen includes standard treatment for treating acute exacerbations, such as corticosteroids (e.g., methylprednisolone) and / or plasma exchange. In some embodiments, the prior treatment regimen includes standard treatment for preventing exacerbations, such as eculizumab (a humanized monoclonal antibody) and / or other immunosuppressants (e.g., azathioprine, rituximab, and mycophenolate mofetil). In some embodiments, the AQP4-IgG positive neuromyelitis optica spectrum disorder has not received non-pharmacological treatment.

[0028] In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered by injection, such as intravenous injection, subcutaneous injection, or intraperitoneal injection; or, the administration method is intravenous infusion.

[0029] In some embodiments, the anti-CD20 antibody is selected from monoclonal antibodies or CD20 binding fragments.

[0030] In some embodiments, the anti-CD20 antibody comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2.

[0031] In some embodiments, the light chain variable region comprises an amino acid sequence that is sequence-identical to SEQ ID NO:1 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or any two of these values ​​(including endpoint values).

[0032] In some embodiments, the heavy chain variable region comprises an amino acid sequence that is sequence-identical to SEQ ID NO:2 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or any two of these values ​​(including endpoint values).

[0033] In some embodiments, the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:1, and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:2.

[0034] In some embodiments, the anti-CD20 antibody comprises a light chain and a heavy chain, the light chain comprising an amino acid sequence as shown in SEQ ID NO:3 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:3, and the heavy chain comprising an amino acid sequence as shown in SEQ ID NO:4 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:4.

[0035] In some embodiments, the light chain comprises an amino acid sequence that is sequence-identical to SEQ ID NO:3 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or any two of these values ​​or a range (including endpoint values).

[0036] In some embodiments, the heavy chain comprises an amino acid sequence that is sequence-identical to SEQ ID NO:4 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or any two of these values ​​or a range (including endpoint values).

[0037] In some embodiments, the anti-CD20 antibody comprises a light chain and a heavy chain, the light chain comprising an amino acid sequence as shown in SEQ ID NO:3, and the heavy chain comprising an amino acid sequence as shown in SEQ ID NO:4.

[0038] In some embodiments, the anti-CD20 antibody comprises two light chains and two heavy chains; wherein each light chain comprises an amino acid sequence as shown in SEQ ID NO:3, and each heavy chain comprises an amino acid sequence as shown in SEQ ID NO:4.

[0039] In some embodiments, the anti-CD20 antibody is BAT4406F, which comprises a light chain and a heavy chain, the light chain comprising the amino acid sequence shown in SEQ ID NO:3, and the heavy chain comprising the amino acid sequence shown in SEQ ID NO:4; and the anti-CD20 antibody has a very low (not exceeding 5%) content of fucose-containing anti-CD20 antibody.

[0040] In some embodiments, the anti-CD20 antibody is BAT4406F, which comprises two light chains and two heavy chains; wherein each light chain comprises an amino acid sequence as shown in SEQ ID NO:3, and each heavy chain comprises an amino acid sequence as shown in SEQ ID NO:4; and the anti-CD20 antibody has a very low (not exceeding 5%) content of fucose-containing anti-CD20 antibody.

[0041] BAT4406F, as described herein, is a fully human monoclonal antibody of the glycosylation-optimized IgG1 subclass described in international patent application PCT / CN2018 / 100008, which is incorporated herein by reference in its entirety.

[0042] In some embodiments, anti-CD20 antibodies or their antigen-binding fragments (e.g., BAT4406F) are produced by cell lines that have knocked out the α-(1,6)fucosyltransferase gene, which can express antibodies with low fucose content (e.g., 0-5%) and enhanced ADCC effect.

[0043] In some embodiments, the glycosylation of the anti-CD20 antibody (e.g., BAT4406F) is characterized by one or more of the following:

[0044] The content of antibodies containing fucose is very low (e.g., 0-5%);

[0045] Low levels of galactose-containing antibodies (e.g., ≤30%);

[0046] Low levels of antibodies containing mannose (e.g., ≤5%);

[0047] High levels of mannose result in low antibody levels (e.g., ≤5%).

[0048] High levels of antibodies containing G0 (e.g., ≥60%).

[0049] In some embodiments, the antibody has a low level of galactose-containing antibody, for example, ≤5%.

[0050] In some embodiments, the antibody has a high level of G0-containing antibodies, for example, ≥80%.

[0051] In some embodiments, the glycosylation site of the antibody is an Asn residue on the heavy chain, such as Asn297 in SEQ ID NO:4.

[0052] In some embodiments, at least about 50% of the anti-CD20 antibody (e.g., BAT4406F) may contain the GO glycan at asparagine residue 297 (Asn297) of SEQ ID NO:4, and / or up to about 10% of the anti-CD20 antibody may contain the Man5 glycan at amino acid residue 297 of SEQ ID NO:4. In some embodiments, less than about 20% of the anti-CD20 antibody contains a fucosylation residue. In some embodiments, less than about 10% of the anti-CD20 antibody contains a fucosylation residue. In some embodiments, less than about 5% of the anti-CD20 antibody contains a fucosylation residue. In some embodiments, less than about 1% of the anti-CD20 antibody contains a fucosylation residue.

[0053] In some embodiments, at least 50% of the anti-CD20 antibodies (e.g., BAT4406F) contain GO glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody. In some embodiments, at least 60% of the anti-CD20 antibodies contain GO glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody. In some embodiments, at least about 65% of the anti-CD20 antibodies may contain GO glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody. In some embodiments, up to about 7% of the anti-CD20 antibodies may contain Man5 glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody. In some embodiments, up to about 5% of the anti-CD20 antibodies may contain Man5 glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody. In some embodiments, up to about 3% of the anti-CD20 antibodies may contain Man5 glycans at the N-glycosylation sites in the heavy chain constant region of the anti-CD20 antibody.

[0054] In some embodiments, no more than about 10% of the oligosaccharides in the Fc region of the anti-CD20 antibody (e.g., BAT4406F) comprise fucose. In some embodiments, no more than about 5% of the oligosaccharides in the Fc region of the anti-CD20 antibody comprise fucose. In some embodiments, no more than about 1% of the oligosaccharides in the Fc region of the anti-CD20 antibody comprise fucose.

[0055] In some embodiments, at least or at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, or any number or range between any two of these values ​​(including endpoint values) of the antiCD20 antibody may contain GO glycans at the N-glycosylation sites of the heavy chain constant region of the antiCD20 antibody. In some embodiments, up to or up to about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or any number or range between any two of these values ​​(including endpoint values) of antiCD20 antibodies may contain any glycan other than the G0 glycan at the N-glycosylation site of the heavy chain constant region of the antiCD20 antibody.

[0056] In some embodiments, at most or at most about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or any number or range between any two of these values, of an anti-CD20 antibody (e.g., BAT4406F) may contain GO-GN glycans at the N-glycosylation sites of the heavy chain constant region of the anti-CD20 antibody. In some embodiments, up to or up to about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any number or range between any two of these values ​​(including endpoint values) of the antiCD20 antibody may contain Man5 glycan at the N-glycosylation site of the heavy chain constant region of the antiCD20 antibody. In some embodiments, up to or up to about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or any number or range between any two of these values ​​(including endpoint values) of antiCD20 antibody may contain G1 glycan at the N-glycosylation site of the heavy chain constant region of the antiCD20 antibody. In some embodiments, up to or up to about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or any number or range between any two of these values ​​(including endpoint values) of antiCD20 antibody may contain G1' glycan at the N-glycosylation site of the heavy chain constant region of the antiCD20 antibody.In some embodiments, up to or up to about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or any number or range between any two of these values ​​(including endpoint values) of antiCD20 antibody may contain G2 glycan at the N-glycosylation site of the heavy chain constant region of the antiCD20 antibody.

[0057] In some embodiments, the fucose-containing anti-CD20 antibody (e.g., BAT4406F) is present in very low amounts. For example, the fucose-containing antibody content of the anti-CD20 antibody is, approximately, or at most, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any number or range between any two of these values. In some embodiments, the anti-CD20 antibody (e.g., BAT4406F) does not contain antibodies with fucose residues (e.g., unfucosylated or defucosylated BAT4406F).

[0058] In some embodiments, the level of the galactose-containing anti-CD20 antibody is low. For example, the level of the galactose-containing antibody is, approximately, or at most, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, or any number or range between any two of these values ​​(including endpoint values).

[0059] In some embodiments, the content of the mannose-containing anti-CD20 antibody is very low. For example, the content of the mannose-containing anti-CD20 antibody is, approximately, or at most 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any number or range between any two of these values ​​(including endpoints).

[0060] In some embodiments, the content of the high-mannose anti-CD20 antibody is very low. For example, the content of the high-mannose anti-CD20 antibody is, approximately, or at most 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any number or range between any two of these values ​​(including endpoints).

[0061] In some implementations, the level of the anti-CD20 antibody containing G0 is high. For example, the level of the anti-CD20 antibody containing G0 is, is approximately, or is at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or any number or range between any two of these values ​​(including endpoint values).

[0062] The anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment can be formulated into a pharmaceutical composition and administered to patients in various forms suitable for the chosen route of administration, such as intravenous (iv), intramuscular, local, or subcutaneous parenteral administration. The dosage of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment can vary depending on the nature of the drug, the degree of internalization at the cell surface, drug transduction and release, the disease being treated, and the patient's condition, such as age, sex, and weight.

[0063] In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered from about 0.5 mg / kg to 20 mg / kg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered from about 0.5 mg / kg to about 15 mg / kg. In some implementations, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, or any number or range between these values. In some embodiments, the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered as a single dose, or weekly, monthly, every two months, every three months, every four months, every five months, every six months, every seven months, every eight months, every nine months, every ten months, every eleven months, or every twelve months, or any number or range between these values. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered as needed due to disease relapse.

[0064] In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 10 mg to about 3000 mg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 10 mg to about 1500 mg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 20 mg to about 1000 mg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 100 mg to about 800 mg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 500 mg to about 790 mg. In some embodiments, the dose of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is from about 710 mg to about 790 mg. In some embodiments, the dosage of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is approximately 10 mg, approximately 20 mg, approximately 50 mg, approximately 100 mg, approximately 150 mg, approximately 200 mg, approximately 250 mg, approximately 300 mg, approximately 350 mg, approximately 400 mg, approximately 450 mg, approximately 500 mg, approximately 600 mg, approximately 700 mg, approximately 750 mg, approximately 800 mg, approximately 900 mg, approximately 1000 mg, or approximately 1100 mg. g, about 1200 mg, about 1300 mg, about 1400 mg, about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg, about 2000 mg, about 2100 mg, about 2200 mg, about 2300 mg, about 2400 mg, about 2500 mg, about 2600 mg, about 2700 mg, about 2800 mg, about 2900 mg, about 3000 mg, or any number or range between any two of these values. In some embodiments, the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered as a single dose, or weekly, monthly, every two months, every three months, every four months, every five months, every six months, every seven months, every eight months, every nine months, every ten months, every eleven months, or every twelve months, or any number or range between any two of these values. In some implementations, the anti-CD20 antibody or its antigen-binding fragment is administered as needed due to disease recurrence.

[0065] In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 20 mg, approximately every four months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 20 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 100 mg, approximately every five months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 100 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 200 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 500 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 500 mg, approximately every five months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 500 mg, approximately every four months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 550 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 600 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 650 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 700 mg, approximately every six months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 750 mg, approximately every four months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 750 mg, approximately every five months. In some embodiments, the anti-CD20 antibody or its antigen-binding fragment is administered at a dose of about 750 mg, approximately every six months.

[0066] In some embodiments, the present invention provides a method of treating NMOSD comprising administering an effective amount of an anti-CD20 antibody (e.g., BAT4406F) to a patient in need, said anti-CD20 antibody being administered at a dose of about 710 mg to about 790 mg (e.g., about 750 mg), as a single dose, or about every six months, or as needed due to disease relapse. In some embodiments, said NMOSD is AQP4-IgG negative NMOSD. In some embodiments, said NMOSD is AQP4-IgG positive NMOSD.

[0067] In some embodiments, the present invention provides a method of treating NMOSD comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered at a dose of about 710 mg to about 790 mg (e.g., about 750 mg), as a single dose, or about every six months, or as needed due to disease recurrence. In some embodiments, said NMOSD is AQP4-IgG negative NMOSD. In some embodiments, said NMOSD is AQP4-IgG positive NMOSD.

[0068] In some embodiments, the present invention provides a method for treating NMOSD, comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered in a single dose of about 750 mg, or approximately every six months. In some embodiments, said NMOSD is AQP4-IgG negative NMOSD. In some embodiments, said NMOSD is AQP4-IgG positive NMOSD.

[0069] In some embodiments, the present invention provides a method for treating AQP4-IgG negative NMOSD, comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered in a dose of about 750 mg, as a single dose or approximately every six months. In some embodiments, the present invention provides a method for treating AQP4-IgG positive NMOSD, comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered in a dose of about 750 mg, as a single dose or approximately every six months.

[0070] In some embodiments, the present invention provides a method for treating AQP4-IgG negative NMOSD, comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered in a dose of about 500 mg, as a single dose or approximately every six months. In some embodiments, the present invention provides a method for treating AQP4-IgG positive NMOSD, comprising administering an effective amount of BAT4406F to a patient in need, said BAT4406F being administered in a dose of about 500 mg, as a single dose or approximately every six months.

[0071] In some implementations, the total number of administrations of the anti-CD20 antibody or its antigen-binding fragment may be approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, or any number or range between any two of these values ​​(including endpoints). For example, the anti-CD20 antibody or its antigen-binding fragment may be administered approximately 4 times.

[0072] In some embodiments, the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered via intravenous infusion. In some embodiments, the duration of intravenous infusion depends on the administration circumstances. In some embodiments, the duration of intravenous infusion is, or approximately, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 14 hours, or any number or range between these values ​​(including endpoints). For example, administration may be performed over approximately 1 hour.

[0073] In some embodiments, the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment is administered in combination with at least one other therapy. For example, the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment may be administered in combination with at least one other therapy for treating the disease described herein (e.g., immunosuppressants, corticosteroids, antimalarial agents, cytotoxic agents, integrin antagonists, cytokine antagonists, or hormones). In some embodiments, the other therapy is selected from systemic steroids, nervous system drugs, musculoskeletal system drugs, digestive and metabolic system drugs, and systemic anti-infectives, or combinations thereof. In some embodiments, the systemic steroids do not include sex hormones and insulin. In some embodiments, the alternative therapy is selected from corticosteroids (e.g., glucocorticoids), antipyretic analgesics, gabapentin, benzodiazepine derivatives, nervous system drugs, combination drugs of antidepressants and tranquilizers, centrally acting muscle relaxants, anxiolytics, anticholinesterases, vitamin B1, digestive and metabolic system drugs, and normal human immunoglobulins, or combinations thereof. In some embodiments, the alternative therapy is a glucocorticoid.

[0074] In some embodiments, the present invention provides pharmaceutical compositions comprising an anti-CD20 antibody (e.g., BAT4406F) or an antigen-binding fragment thereof as described herein. For example, such a composition may comprise 0.1% or more of an anti-CD20 antibody (e.g., BAT4406F) or an antigen-binding fragment thereof. The percentage of the anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment thereof may vary and may range from about 2% to about 90% by weight of a given dosage form. The amount of anti-CD20 antibody (e.g., BAT4406F) or its antigen-binding fragment in such therapeutically useful compositions allows for the administration of an effective amount.

[0075] Examples of pharmaceutical compositions suitable for injection or infusion may include sterile aqueous solutions or dispersions in pharmaceutically acceptable liquid carriers or excipients, or sterile powders containing the active ingredient suitable for immediate preparation of sterile injectable or infusionable solutions or dispersions, optionally encapsulated in liposomes. Other forms of pharmaceutical compositions include topical formulations such as gels, ointments, creams, lotions, or transdermal patches. Pharmaceutical compositions, in addition to those mentioned herein, also include suitable pharmaceutically acceptable carriers known in the art; see, for example: Remington, The Science and Practice of Pharmacy, 20th edition, 2000, Lippincott Williams and Wilkins (Editors: Gennaro, AR, et al.).

[0076] On the other hand, the present invention provides the use of an effective amount of an anti-CD20 antibody or its antigen-binding fragment in the preparation of a medicament for treating a disease; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0077] The effective amount is approximately 710 mg to approximately 790 mg per dose, for example, approximately 750 mg.

[0078] In the aforementioned uses, the disease, the CD20 antibody or its antigen-binding fragment, the administration method, etc., are as described above and will not be repeated here.

[0079] On the other hand, the present invention also provides the use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the treatment of a disease; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0080] The effective amount is approximately 710 mg to approximately 790 mg per dose, for example, approximately 750 mg.

[0081] In the aforementioned uses, the disease, the CD20 antibody or its antigen-binding fragment, the administration method, etc., are as described above and will not be repeated here.

[0082] On the other hand, the present invention provides the use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the preparation of a medicament for treating AQP4-IgG negative neuromyelitis optica spectrum disorders; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0083] The effective dose is approximately 10 mg to approximately 3000 mg per dose.

[0084] In the aforementioned applications, the CD20 antibody or its antigen-binding fragment, administration method, etc., are as described above and will not be repeated here.

[0085] On the other hand, the present invention also provides the use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the treatment of AQP4-IgG negative neuromyelitis optica spectrum disorders; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0086] The effective dose is approximately 10 mg to approximately 3000 mg per dose.

[0087] In the aforementioned applications, the CD20 antibody or its antigen-binding fragment, administration method, etc., are as described above and will not be repeated here.

[0088] On the other hand, the present invention provides the use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the preparation of a medicament for treating AQP4-IgG positive neuromyelitis optica spectrum disorders; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0089] The effective dose is approximately 10 mg to approximately 3000 mg per dose.

[0090] In the aforementioned applications, the CD20 antibody or its antigen-binding fragment, administration method, etc., are as described above and will not be repeated here.

[0091] On the other hand, the present invention also provides the use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the treatment of AQP4-IgG positive neuromyelitis optica spectrum disorders; wherein the anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; the heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2;

[0092] The effective dose is approximately 10 mg to approximately 3000 mg per dose.

[0093] In the aforementioned applications, the CD20 antibody or its antigen-binding fragment, administration method, etc., are as described above and will not be repeated here. Attached Figure Description

[0094] Figure 1A is a constant coordinate graph of the mean drug-time curve (Mean±SD) after intravenous infusion of BAT4406F in the subjects.

[0095] Figure 1B is a semi-logarithmic graph of the mean drug-time curve (Mean±SD) of the subjects after intravenous infusion of BAT4406F.

[0096] Figure 2 shows the average CD19 of the subjects. +B lymphocyte count-time semi-logarithmic curve (logarithmic scale ordinate, Mean±SD). Detailed Implementation

[0097] definition

[0098] As used herein, unless otherwise stated, the following definitions should apply.

[0099] As used herein, unless otherwise stated, the singular forms “a,” “an,” and “the” include the plural forms. Thus, for example, reference to “a compound” includes multiple compounds.

[0100] As used herein, “about” will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. In some embodiments, “about” will mean up to ±10% or ±5%, or ±1%, of a particular term. “about x” includes “x”.

[0101] "Amino acid" refers to α-aminocarboxylic acid, which can be encoded by nucleic acids directly or in precursor form. A single amino acid is encoded by a nucleic acid consisting of three nucleotides (so-called codons or base triplets). Each amino acid is encoded by at least one codon. The fact that the same amino acid is encoded by different codons is called "degeneracy of the genetic code." As used in this application, the term "amino acid" refers to naturally occurring α-aminocarboxylic acids, including alanine (three-letter code: Ala, or one-letter code: A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine ​​(Cys, C), glutamine (Gln, Q), glutamic acid (Glu, E), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), and valine (Val, V).

[0102] The term “antibody” is used in a broad sense and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (such as bispecific antibodies), and antibody fragments, provided they exhibit the desired biological activity.

[0103] An "antibody fragment" comprises a portion of a full-length antibody, typically containing its antigen-binding region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; dimers; linear antibodies; single-chain antibodies; and multispecific antibodies formed from antibody fragments.

[0104] Monoclonal antibodies (mAbs) are antibodies prepared from the same immune cells, which are all clones of a single parent cell. Monoclonal antibodies can have monovalent affinity because they bind to the same epitope (the part of the antigen that the antibody recognizes). In contrast, polyclonal antibodies bind to multiple epitopes and are typically secreted by several different plasma cells. Monoclonal antibodies can also be modified into bispecific monoclonal antibodies by adding a therapeutic target to two epitopes. Monoclonal antibodies can be prepared using hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0105] As used herein, the term "comprising" is intended to indicate that a composition and method includes the listed elements but does not exclude other elements. When used to define compositions and methods, "consisting substantially of" should mean excluding other elements that are of any significant importance to the composition. For example, a composition consisting substantially of the elements defined herein does not exclude other elements that do not substantially affect the essential and novel features of the claimed invention. "Constitutes" should mean excluding other ingredients in trace amounts and the substantial method steps described herein. Embodiments defined by each of these transitional terms are within the scope of this disclosure.

[0106] The terms "patient" and "subject" are used interchangeably to refer to mammals, including humans and non-human mammals. In some embodiments, the patient is a human. In some embodiments, the patient is a non-human mammal, such as a wild, domesticated, or farm animal. In other embodiments, the patient is a dog, cat, mouse, rat, rabbit, guinea pig, or primate such as a cynomolgus monkey.

[0107] "Treating" a patient's disease means (1) preventing the disease from occurring in patients who are susceptible or who have not yet shown symptoms of the disease; (2) suppressing the disease or its symptoms or preventing its development; or (3) reducing or eliminating the disease or its symptoms.

[0108] "Effective amount" refers to the amount of an active compound or agent that elicits a biological or medical response in a tissue, system, animal, individual, or human. The effective amount is sought by researchers, veterinarians, physicians, or other clinicians, including for the treatment of disease.

[0109] "Administration" and "dosage" are used interchangeably and can be accomplished by injection, infusion, parenteral administration, intravenous administration, mucosal administration, sublingual administration, intramuscular administration, intradermal administration, intranasal administration, intraperitoneal administration, intra-arterial administration, subcutaneous absorption, or by any method in combination with other known techniques. "Administration" can also refer to the prescription of a composition with instructions for administration using one or more of the aforementioned techniques. In some embodiments, administration is systemic.

[0110] As used herein, the phrase "need" means that a patient has been identified as requiring a specific method or treatment. In some implementations, this identification can be made by any diagnostic means. In any of the methods and treatments described herein, the patient may require these methods and treatments.

[0111] In the following examples, AUC (0-inf) AUC represents the area under the concentration-time curve from time 0 to infinity. (0-t) C represents the area under the concentration-time curve from time 0 to the last measurable time point. max T represents the maximum plasma concentration. max t represents the time to reach maximum plasma concentration. 1 / 2 CL represents the time required for plasma concentration to halve. t V represents plasma clearance rate. d The apparent volume is represented by MRT, which represents the mean retention time.

[0112] "Non-pharmacological treatment" refers to the use of physical and chemical methods or means other than drugs to treat diseases.

[0113] "Number of occurrences" refers to the number of times a current type of adverse event occurs. For example, if a subject experiences 3 adverse events, it is counted as 3 occurrences.

[0114] All publications are incorporated herein by reference in their entirety for all purposes.

[0115] The following specific embodiments further illustrate the technical solution of the present invention. These specific embodiments do not represent a limitation on the scope of protection of the present invention. Non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the scope of protection of the present invention.

[0116] Example: Clinical study of BAT4406F

[0117] This study is a single-center, open-label, dose-escalation phase I clinical trial evaluating the safety, tolerability, pharmacokinetics (PK), immunogenicity, and preliminary efficacy of BAT4406F in patients with neuromyelitis optica spectrum disorder (NMOSD).

[0118] Research objective:

[0119] Main research objective:

[0120] 1. To evaluate the safety and tolerability of dose escalation of BAT4406F in NMOSD patients to determine the maximum tolerated dose (MTD) and dose-limiting toxicities (DLT);

[0121] 2. Evaluate the p-axis characteristics of BAT4406F;

[0122] 3. Study the pharmacodynamic characteristics of BAT4406F.

[0123] Secondary research objective:

[0124] 1. To investigate the immunogenicity of BAT4406F;

[0125] 2. Preliminary evaluation of the efficacy of BAT4406F in NMOSD patients.

[0126] In this study, DLT was defined as any of the following adverse events (AEs) occurring within 28 days of administration and related to the study treatment, graded according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0:

[0127] Grade 3 or higher drug-related non-hematological toxicity;

[0128] Grade 4 neutropenia lasting more than 7 days;

[0129] Grade 3 fever and neutropenia requiring intravenous antibiotic treatment;

[0130] Grade 4 thrombocytopenia and other hematological toxicities

[0131] However, grade 3 or 4 infusion-related reactions are not classified as DLT.

[0132] Research Methods:

[0133] A "3+3" dose escalation principle was adopted, with 5 fixed-dose groups: 20 mg, 100 mg, 200 mg, 500 mg, and 750 mg. A single dose was administered, and the observation period was 6 months. The diuretic tolerance (DLT) observation period was 28 days after administration. If no DLT was observed within 28 days, treatment continued in the next higher-dose group until the mean time to treatment (MTD) was reached or the study termination criteria were met.

[0134] Selection criteria:

[0135] 1. Meets the NMOSD diagnostic criteria established by the International NMO Diagnostic Panel (IPND) in 2015;

[0136] 2. Patients aged ≥18 and ≤65 years, regardless of gender;

[0137] 3. Have experienced at least two relapses within the two years prior to screening, or at least one relapse within the year prior to screening;

[0138] 4. Discontinue immunosuppressants such as azathioprine, tacrolimus, and mycophenolate mofetil within 28 days prior to baseline.

[0139] 5. If glucocorticoids are used during screening, the dose should be 30 mg or less of the prednisone equivalent;

[0140] 6. Extended Disability Status Scale (EDSS) score ≤ 6.

[0141] Exclusion criteria:

[0142] 1. Has received any monoclonal antibody treatment within 6 months prior to administration;

[0143] 2. Those who have previously received anti-CD20 monoclonal antibody therapy;

[0144] 3. Received intravenous immunoglobulin within one month prior to screening;

[0145] 4. Had plasma exchange within one month prior to screening;

[0146] 5. Received a live vaccine within 4 weeks prior to baseline;

[0147] 6. Participated in another clinical study and the baseline time was less than 1 month after treatment with the investigational drug or 5 half-lives of the drug (whichever is longer);

[0148] 7. Those with a history of allergy to monoclonal antibodies; those with an allergic constitution (a history of severe allergic reactions to a certain drug or food);

[0149] 8. Abnormal liver and kidney function, and bone marrow reserve:

[0150] a. Alanine aminotransferase (ALT) ≥ 2.5 × upper limit of normal (ULN) and / or aspartate aminotransferase (AST) ≥ 2.5 × ULN and / or total bilirubin (TBIL) ≥ 1.5 × ULN;

[0151] b. Serum creatinine ≥ 1.5 × ULN, creatinine clearance ≤ 60 mL / min (calculated using the Cockcroft formula);

[0152] c. Serum neutrophil count <1.5×10 9 / L and / or platelet count <75×10 9 Patients with / L;

[0153] 9. History of human immunodeficiency virus (HIV) infection or HIV positivity at enrollment screening; history of hepatitis B and / or hepatitis C infection or positive hepatitis B surface antigen (HBsAg) at enrollment screening [or simultaneously meeting the following three criteria: positive hepatitis B core antibody, negative hepatitis B surface antibody, or hepatitis B virus deoxynucleic acid (HBV DNA) quantification exceeding the normal range]; or positive hepatitis C virus (HCV) antibody; positive Treponema pallidum antibody at enrollment;

[0154] 10. Subjects meet any of the following criteria associated with latent or active tuberculosis (TB) infection:

[0155] a. A history of active TB within ≤3 years prior to screening (but if it is >3 years and there is a documented record of completing adequate treatment, enrollment is also possible);

[0156] b. Signs or symptoms suggestive of active TB are present during the medical history and / or physical examination during screening;

[0157] c. Recent close contact with someone with active TB;

[0158] d. A positive T-cell test result for tuberculosis infection at screening is acceptable. If a subject's first T-cell test result for tuberculosis infection is indeterminate, that subject will be excluded from the study. Exceptions may be made for subjects who have completed appropriate standard treatment for latent TB prior to screening and have no other risk factors, imaging findings, or specific indications supporting latent or active TB. Investigators must assess the tuberculosis (including extrapulmonary) risk with a qualified infectious disease physician and discuss the decision with the sponsor.

[0159] 11. Developing clinical symptoms or signs consistent with COVID-19 infection (e.g., fever, dry cough, difficulty breathing, sore throat, and fatigue) within 4 weeks prior to or during screening, or being confirmed to have COVID-19 infection by appropriate laboratory testing (as determined by the investigator or in accordance with local regulations). If confirmed to have COVID-19 infection before screening, recovery from the infection must be demonstrated by appropriate laboratory testing.

[0160] 12. History of infection deemed unsuitable for participation by other researchers;

[0161] 13. Patients with acute respiratory failure and pulmonary interstitial infiltration and edema visible on chest CT.

[0162] 14. Patients with a clear history of heart disease (e.g., angina pectoris, atrial flutter and atrial fibrillation, conduction block and other arrhythmias or heart failure); for patients with first-degree atrioventricular block, the researcher shall make a comprehensive judgment based on the patient's medical history.

[0163] 15. Those who also have malignant tumors;

[0164] 16. History of mental disorder;

[0165] 17. Suffering from other neurological disorders;

[0166] 18. Patients with metabolic, hematological, renal, hepatic, pulmonary, neurological, endocrine, cardiac, infectious, or gastrointestinal diseases that researchers believe pose an unacceptable risk to the patient or may affect NMOSD assessment.

[0167] 19. History of drug or alcohol abuse (14 units of alcohol per week: 1 unit = 285 mL of beer, 25 mL of spirits, or 125 mL of wine) within 6 months prior to screening;

[0168] 20. Pregnant or breastfeeding women, and female subjects with positive pregnancy tests at screening or baseline.

[0169] Administration method:

[0170] The medication was administered once via intravenous infusion. All subjects received corticosteroids and antihistamines at least 30 minutes prior to infusion to reduce the risk of infusion-related reactions. The initial infusion rate was approximately 12 mL / h. If no infusion reaction occurred, the rate was increased by 12 mL / h every half hour until a maximum infusion rate of 96 mL / h was reached.

[0171] Evaluation criteria:

[0172] Tolerability and safety:

[0173] ●Evaluate whether DLT occurs or MTD is reached during dose ramp-up;

[0174] ● Vital signs (heart rate / pulse rate, respiration, blood pressure, body temperature), physical examination, laboratory tests (urinalysis, complete blood count, blood biochemistry, coagulation function, AQP4-IgG, etc.), 12-lead electrocardiogram (ECG), adverse events (AEs), injection site examination, etc.

[0175] Pharmacokinetics (PK):

[0176] ●C max Peak time (T) max Terminal elimination half-life (T) 1 / 2 Total clearance (CL), apparent volume of distribution (V) d ), mean residence time from 0 to infinity (MRT) 0-inf The area under the blood drug concentration-time curve (AUC) from time 0 to time t. 0-t The area under the blood drug concentration-time curve (AUC) from 0 to infinity 0-inf )wait.

[0177] Pharmacodynamics:

[0178] ●CD19 levels in peripheral blood at various follow-up time points before and after medication administration + B lymphocyte percentage / count

[0179] Among them: CD19 +B lymphocyte count = CD19 + B lymphocyte percentage × lymphocyte count, unit: 10 7 / L. When CD19 + B lymphocyte count less than 10 cells / μL (i.e., 10 7 When the cell count reaches 1 / L, it is considered as B cell depletion.

[0180] Immunogenicity:

[0181] ● Positive rate of anti-drug antibody (ADA);

[0182] ●Titer of ADA-positive samples;

[0183] ● Samples that test positive for ADA will be further tested to determine if they contain neutralizing antibodies (NAb).

[0184] Therapeutic effects:

[0185] ● The proportion of subjects who relapsed during the observation period, the annual relapse rate, and the time to the first relapse after administration.

[0186] ● Change in EDSS score from baseline.

[0187] ●MRI scans show changes compared to baseline.

[0188] ● Changes in ophthalmological examinations.

[0189] result:

[0190] A total of 15 subjects were enrolled, including 14 females and 1 male. The subjects' ages ranged from 26 to 64 years, with a mean age ± SD of 42.13 ± 10.66 years. Nine subjects were aged between 18 and 40 years, and six were aged between 41 and 65 years. All subjects had a history of NMOSD recurrence. At baseline, 7 subjects (46.7%) were AQP4-IgG positive, and 8 subjects (53.3%) were AQP4-IgG negative.

[0191] Three subjects were enrolled in each of the 20mg, 100mg, 200mg, 500mg, and 750mg groups, respectively. All 15 subjects received BAT4406F at the prescribed dose, with 100% dosage adherence. Thirteen subjects (86.7%) completed the study without relapse, while two subjects (13.3%) withdrew early due to disease relapse during the observation period (one from the 20mg group and one from the 200mg group).

[0192] Previous drug treatment for NMOSD

[0193] In this study, 13 subjects (86.7%) had received 119 previous treatments with NMOSD, namely 20 mg group (3 cases, 100%), 200 mg group (2 cases, 66.7%), 200 mg group (2 cases, 66.7%), 500 mg group (3 cases, 100%), and 750 mg group (3 cases, 100%), 47 treatments.

[0194] According to the ATC1 classification, previous drug treatments for NMOSD included: systemic hormones, excluding sex hormones and insulin (12 cases, 80.0%), 92 treatments; antitumor and immunomodulatory drugs (8 cases, 53.3%), 11 treatments; and neurological drugs (7 cases, 46.7%), 16 treatments.

[0195] According to the ATC4 classification, prior medical treatment for NMOSD included: glucocorticoids (12 cases, 80.0%) 92 times, selective immunosuppressants (5 cases, 33%) 8 times, gabapentin (5 cases, 33%) 7 times, other nervous system drugs (3 cases, 20.0%) 3 times, other immunosuppressants (3 cases, 20.0%) 3 times, other antipyretic analgesics (2 cases, 13.3%) 3 times, combination therapy of antidepressants and benzodiazepines (2 cases, 13.3%) 2 times, and adamantane derivatives (1 case, 6.7%) 1 time. No subjects received prior non-pharmacological treatment for NMOSD.

[0196] Drug treatment during the NMOSD trial

[0197] Table 1 summarizes the ATC classification of NMOSD concomitant medications during the trial period. 15 subjects (100%) received 101 doses of NMOSD concomitant medication treatment, namely 20 mg group (3 cases, 100%), 27 doses; 100 mg group (3 cases, 100%), 24 doses; 200 mg group (3 cases, 100%), 20 doses; 500 mg group (3 cases, 100%), 17 doses; and 750 mg group (3 cases, 100%), 13 doses.

[0198] According to the ATC1 classification, concomitant medications for NMOSD included: systemic hormones (excluding sex hormones and insulin) 65 times (14 cases, 93.3%), nervous system medications 28 times (12 cases, 80.0%), musculoskeletal system medications 5 times (2 cases, 13.3%), digestive and metabolic system medications 2 times (2 cases, 13.3%), and systemic anti-infectives 1 time (1 case, 6.7%).

[0199] According to the ATC4 classification, concomitant medications for NMOSD included: glucocorticoids (14 cases, 93.3%) 65 doses, other antipyretic analgesics (7 cases, 46.7%) 11 doses, gabapentin (6 cases, 40.0%) 8 doses, benzodiazepine derivatives (2 cases, 13.3%) 3 doses, other neurological drugs (2 cases, 13.3%) 2 doses, and combination therapies of antidepressants and tranquilizers (2 cases, ...). The following medications were administered: 2 doses of NMOSD (13.3%), 5 doses of other centrally acting muscle relaxants (2 doses, 13.3%), 1 dose of other anti-anxiety medications (1 dose, 6.7%), 1 dose of anticholinesterase inhibitors (1 dose, 6.7%), 1 dose of vitamin B1 alone (1 dose, 6.7%), 1 dose of various digestive and metabolic system medications (1 dose, 6.7%), and 1 dose of normal human immunoglobulin (1 dose, 6.7%). No subjects received NMOSD accompanied by non-pharmacological treatment.

[0200] Table 1. ATC classification of NMOSD concomitant medications during the trial.

[0201] This table only lists data with a total of ≥2 instances.

[0202] Tolerability and safety

[0203] After administration of BAT4406F 20mg to 750mg, all 15 subjects completed the 28-day DLT observation period. None of the subjects experienced DLT, and the MTD of BAT4406F was not reached.

[0204] Regarding safety, a total of 120 adverse events (AEs) occurred in 14 subjects (93.3%), of which 56 adverse reactions (ADRs) occurred in 13 subjects (86.7%). Most AEs resulted in recovery, remission, or stabilization. No subjects experienced ADRs of severity ≥ grade 3, serious adverse reactions, AEs leading to withdrawal, or ADRs leading to withdrawal.

[0205] The vast majority of subjects experienced grade 1 or 2 adverse events (AEs). Only one subject (in the 100 mg group) experienced a grade 3 AE (PT: osteonecrosis), meeting the criteria for a serious adverse event (SAE). This subject was diagnosed with avascular necrosis of the femoral head prior to screening. On day 70 after administration, this subject was hospitalized for hip replacement surgery. This SAE was deemed unlikely to be related to the study drug. No grade 4 or 5 AEs occurred.

[0206] According to PT classification, ADRs with a combined incidence >10% (occurring in at least 2 subjects) included: fever [4 cases (26.7%), 4 occurrences, including 1 case each in the 100mg and 750mg groups, and 2 cases in the 200mg group], increased heart rate [3 cases (20.0%), 3 occurrences, including 1 case in the 500mg group and 2 cases in the 750mg group], feeling cold [3 cases (20.0%), 3 occurrences, including 1 case in the 100mg group and 2 cases in the 200mg group], urinary tract infection [3 cases (20.0%), 5 occurrences, including 1 case in the 20mg group, 3 occurrences, and 1 case each in the 100mg and 200mg groups], and increased blood pressure [2 cases (13.3%), 3 occurrences, including 1 case in the 500mg group and 2 cases in the 750mg group]. [Examples of cases: 1 case in the 750mg group, 1 instance], elevated blood lactate dehydrogenase [2 cases (13.3%), 2 instances, including 1 case each in the 500mg and 750mg groups], elevated fibrin D-dimer [2 cases (13.3%), 2 instances, including 1 case each in the 20mg and 500mg groups], chest discomfort [2 cases (13.3%), 2 instances, in the 750mg group], fever [2 cases (13.3%), 2 instances, including 1 case each in the 100mg and 750mg groups], shortness of breath [2 cases (13.3%), 2 instances, including 1 case each in the 500mg and 750mg groups], allergic dermatitis [2 cases (13.3%), 2 instances, including 1 case each in the 200mg and 500mg groups].

[0207] A total of 4 subjects (26.7%) experienced 11 adverse events (AEs) related to infusion reactions (all ADRs), all of which were grade 1 or 2. The AEs related to infusion reactions included: fever [2 cases (13.3%), 2 occurrences, including 1 case and 1 occurrence in the 100mg group and 1 occurrence in the 750mg group], allergic dermatitis [2 cases (13.3%), 2 occurrences, including 1 case and 1 occurrence in the 200mg group and 1 occurrence in the 500mg group], and chills [1 case (6.7%), 1 occurrence, in the 750mg group]. [Group], feeling cold [1 case (6.7%) 1 time, 200mg group], rash [1 case (6.7%) 1 time, 100mg group], dry throat [1 case (6.7%) 1 time, 200mg group], shortness of breath [1 case (6.7%) 1 time, 500mg group], elevated blood pressure [1 case (6.7%) 1 time, 500mg group], elevated body temperature [1 case (6.7%) 1 time, 200mg group].

[0208] The incidence of adverse events (AEs) related to infusion reactions was comparable across different dose groups.

[0209] A small number of subjects had clinically significant abnormalities in vital signs, physical examination, and ECG results. These abnormalities were related to the subject's medical history, concomitant medications, and / or the study drug.

[0210] Four subjects had normal ECG results at baseline, but developed abnormalities after receiving the medication (including sinus bradycardia, counterclockwise rotation, frequent premature atrial contractions, sinus arrhythmia, and T wave changes), which were determined to be related to the subjects' medical history or adverse events.

[0211] Overall, the safety of BAT4406F is controllable and its tolerance is good.

[0212] Pharmacokinetics

[0213] The mean plasma concentration-time curves (mean plasma concentration-time curves) after intravenous infusion of different doses of BAT4406F in the subjects are shown in Figures 1A and 1B. As can be seen from Figures 1A and 1B, the inflection point of the BAT4406F plasma concentration curve appears approximately 100 hours after administration, indicating that drug distribution mainly occurs around 100 hours after administration; that is, the period from the start of administration to 100 hours is primarily the distribution phase, while the elimination phase is dominant from 100 hours to the end. The 20mg group also shows a second inflection point (approximately 900 hours after administration). The elimination at the end of the plasma concentration curves in the 100mg and 200mg groups shows signs of acceleration, similar to target-mediated drug disposition in biomolecular drugs. The inter-individual coefficients of variation for plasma concentrations at each time point in each dose group were mostly within the range of 30%.

[0214] Pharmacokinetic parameters are shown in Table 2. It can be seen that as the BAT4406F dose increases, T... max The median ranges from 7.00h to 10.90h, T max The median lengthened with increasing BAT4406F dose. Similarly, C... max AUC 0-t and AUC 0-inf All showed an increasing trend with increasing BAT4406F dosage. CL t , λ Z and V d While the mean decreases, the average T 1 / 2 The extension indicates that the elimination of BAT4406F is slower.

[0215] C corrected for dosage max AUC 0-t AUC 0-inf The mean values ​​ranged from 0.27 h / L to 0.49 h / L, 59.52 h / L to 197.59 h / L, and 62.01 h / L to 197.67 h / L, respectively, showing an increasing trend with increasing BAT4406F dosage, indicating that BAT4406F may exhibit nonlinear absorption. Power model analysis showed that the AUC... 0-t and AUC 0-inf Its 95% CI did not fall within the linear judgment interval, C maxThe slope and its 95% CI only partially fall within the linearity judgment interval, suggesting that the overall PK characteristics of BAT4406F are nonlinear in the range of 20 mg to 750 mg.

[0216] Table 2. Summary of pharmacokinetic parameters after intravenous infusion of different doses of BAT4406F in subjects (Mean ± SD, N = 3) Note: Compared with the 20mg group a P<0.05, aa P<0.01; compared with the 100mg group, b P<0.05, bb P<0.01; compared with the 200mg group, c P<0.05, cc P<0.01; compared with the 500mg group, d P<0.05, dd P<0.01.

[0217] Pharmacodynamics

[0218] The average CD19 of subjects in each dose group + The B lymphocyte count-time curve is shown in Figure 2. At baseline, CD19 counts in the 20mg, 100mg, 200mg, 500mg, and 750mg groups were... + The mean ± standard deviation of B lymphocyte counts were (7.17±7.56)×10 / μL, (20.98±13.98)×10 / μL, (10.20±13.86)×10 / μL, (36.29±9.85)×10 / μL, and (29.24±25.88)×10 / μL, respectively. After subjects received intravenous infusions of 20 mg, 100 mg, 200 mg, 500 mg, and 750 mg of BAT4406F, respectively, CD19... + B lymphocyte count decreases rapidly, then remains stable for a period of time before CD19... + B lymphocyte counts showed a slow increase. By day 180, CD19 counts in the 20mg, 100mg, 200mg, 500mg, and 750mg groups were [data missing]. +The mean ± standard deviation of B lymphocyte counts were (4.07±6.35)×10 / μL, (4.25±4.14)×10 / μL, (0.85±0.61)×10 / μL, (0.13±0.15)×10 / μL, and (0.03±0.02)×10 / μL, respectively. The changes from baseline were (-3.10±3.03)×10 / μL, (-16.73±10.47)×10 / μL, (-9.35±13.37)×10 / μL, (-36.17±9.99)×10 / μL, and (-29.22±25.90)×10 / μL, respectively, with change rates from baseline of -5.63±99.18 (%), -79.95±10.14 (%), -82.93±12.09 (%), -99.54±0.59 (%), and -99.75±0.29 (%), respectively.

[0219] Overall, the higher the dose level, the higher the CD19 concentration. + The greater the percentage change in B lymphocyte count compared to baseline, the better. For example, on day 4 after administration, B lymphocyte counts in the 500 and 750 mg groups had decreased to very low levels (mean values ​​of 0.06 and 0.07 × 10⁹ / μL, respectively). Furthermore, the duration of B cell clearance was dose-dependent. The CD19 count in the 20 mg group... + B lymphocyte counts began to rise on day 124, while the 100mg group began to recover on day 152, later than the 20mg group; the B lymphocyte count in the 200mg group recovered on day 180. Both 500mg and 750mg could induce CD19... + B lymphocytes remained at low levels throughout the 6-month observation period (count less than 10 / μL, and B lymphocyte levels in the 750 mg group were lower than those in the 500 mg group after administration).

[0220] The above results indicate that a single dose of BAT4406F has a significant effect on scavenging B cells and can inhibit CD19. + B lymphocytes remained at a low level. The duration of B cell clearance was dose-dependent with increasing BAT4406F dosage.

[0221] In the 20mg, 100mg, 200mg, and 500mg groups, subjects with positive or negative baseline AQP4-IgG results did not experience any change in their AQP4-IgG status during the study period. In the 750mg group, one subject had a negative AQP4-IgG result at baseline, but became positive at follow-up stages D124 and D152 after administration, and became negative at follow-up stage (D180) / early withdrawal. Another subject had a positive AQP4-IgG result at baseline, but became negative at follow-up stages (D124, D180 / early withdrawal).

[0222] Subjects who were AQP4-IgG positive or negative at baseline were included, and their AQP4-IgG status and titer (if positive) were assessed during the study. Results showed that among subjects who were AQP4-IgG positive at baseline, one subject became AQP4-IgG negative after administration of BAT4406F, while the other subjects showed no significant trend in AQP4-IgG titer changes after administration. Two relapsed subjects were AQP4-IgG positive at baseline, but at the time of NMOSD relapse, one subject's AQP4-IgG titer increased compared to baseline, and the other subject's AQP4-IgG titer decreased. All subjects who were AQP4-IgG negative at baseline remained negative during the study, and no one relapsed during the study period.

[0223] therapeutic effect

[0224] During the 6-month observation period, 13 subjects (86.7%) did not experience a relapse of NMOSD, while 2 subjects (13.3%) experienced a relapse around day 40, one in the 20 mg group and one in the 200 mg group. Both of these relapsed subjects discontinued corticosteroids shortly after administration (day 29 and day 4, respectively), therefore their relapse may be related to premature discontinuation of corticosteroids.

[0225] Table 3 summarizes the EDSS scores of subjects in each dosage group of this study. On day 99, the mean changes in EDSS scores from baseline for the 20mg, 100mg, 200mg, 500mg, and 750mg groups were 0.00, 0.00, 0.00, -0.17, and -1.00, respectively, with the 500mg and 750mg groups showing a decrease in mean EDSS scores from baseline. On day 180 / early withdrawal, the mean changes in EDSS scores from baseline were 0.83, -1.00, 0.00, -1.25, and -0.50, respectively, with the 100mg, 500mg, and 750mg groups showing a decrease in mean EDSS scores from baseline (see Table 3).

[0226] Table 3 EDSS scores of subjects in each dose group

[0227] After drug administration, the results of optic nerve, brain, and spinal cord MRI examinations in all dosage groups showed no significant changes compared to baseline. A small number of subjects experienced a change in spinal cord MRI results from clinically significant abnormalities to normal, which may be related to the efficacy of the drug; some subjects experienced a change in ophthalmological examination results from clinically significant abnormalities (related to the primary disease NMOSD) to normal, which may also be related to the efficacy of the drug.

[0228] Immunogenicity

[0229] In this study, 3 subjects tested positive for ADA (1 in the 20mg group and 2 in the 500mg group), with a positive rate of 20% (3 / 15). All 3 subjects who tested positive for ADA had negative neutralizing antibodies (NAb).

[0230] The ADA positivity rates at each time point are shown in Table 4. The ADA positivity rates before administration, and at 168h, 336h, 672h, 1008h, 1680h, 2352h, and D180 after administration were 13.33% (2 / 15), 14.29% (2 / 14), 6.67% (1 / 15), 7.24% (1 / 14), 9.09% (1 / 11), 9.09% (1 / 11), 9.09% (1 / 11), and 0.00 (0 / 12), respectively. The ADA positivity rate generally decreased over time. Specifically, one subject in each of the 20mg and 500mg groups tested positive for ADA before administration, and one subject in the 500mg group tested positive for ADA 168h after administration.

[0231] Table 4. Statistics on ADA positivity rates at different time points

[0232] Table 5 shows the subjects with ADA positivity and antibody titers. One subject in each of the 20mg and 500mg groups was ADA positivity (pre-existing antibody) before administration, with antibody titers of 1:50 and 1:200, respectively. These two subjects had not previously used any anti-CD20 antibodies or other biologics, and their ADA positivity is unlikely to be associated with NMOSD relapse; the potential cause requires further investigation. Another subject in the 500mg group was ADA positivity only on day 8, with an antibody titer of 1:50; this was transient ADA and had no clinical significance. No significant impact was observed on the safety, pharmacokinetics, pharmacodynamics, and efficacy of BAT4406F injection in NMOSD patients.

[0233] Table 5. ADA-positive subjects and antibody titers

[0234] In this study, BAT4406F demonstrated good safety and tolerability in NMOSD patients. No subjects experienced DLT within the study dose range, indicating that it was safe and well-tolerated. BAT4406F exhibited nonlinear pharmacokinetics, possessing rapid and sustained B-cell depletion activity in a dose-dependent manner, and also showed low immunogenicity. Most subjects treated with BAT4406F were relapse-free, suggesting its efficacy in reducing the risk of NMOSD relapse.

Claims

1. A method of treating a disease, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2; in, The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 710 mg to approximately 790 mg.

2. The method according to claim 1, wherein, The diseases are those associated with CD20 expression, such as oncogenic and immune diseases; the oncogenic diseases include B-cell lymphomas, which include Hodgkin's lymphoma and non-Hodgkin's lymphoma; the immune diseases include multiple sclerosis, immune thrombocytopenic purpura, neuromyelitis optica, myasthenia gravis, rheumatoid arthritis, psoriasis, and psoriatic arthritis; or The diseases mentioned are neuromyelitis optica spectrum disorders (NMOSD), non-Hodgkin's lymphoma (NHL), multiple sclerosis (MS), immune thrombocytopenic purpura (ITP), rheumatoid arthritis (RA), Wechsler's granulomatosis (WG), microscopic polyangiitis (MPA), lupus nephritis, systemic lupus erythematosus, and chronic lymphocytic leukemia (CLL); wherein, the neuromyelitis optica spectrum disorders include AQP4-IgG negative neuromyelitis optica spectrum disorders and AQP4-IgG positive neuromyelitis optica spectrum disorders.

3. The method according to claim 1 or 2, wherein, The disease is refractory to at least one prior treatment regimen or has relapsed after at least one prior treatment regimen.

4. The method according to any one of claims 1-3, wherein, The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 750 mg.

5. A method for treating AQP4-IgG negative neuromyelitis optica spectrum disorder, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2; in, The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 10 mg to approximately 3000 mg.

6. A method for treating AQP4-IgG positive neuromyelitis optica spectrum disorder, comprising administering to a patient in need an effective amount of an anti-CD20 antibody or an antigen-binding fragment thereof, said anti-CD20 antibody or antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, said light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; said heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:2; in, The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 10 mg to approximately 3000 mg.

7. The method according to claim 5 or 6, wherein, The dosage of the anti-CD20 antibody or its antigen-binding fragment is about 10 mg to about 1500 mg, or about 20 mg to about 1000 mg, or about 100 mg to about 800 mg, or about 500 mg to about 790 mg, or about 550 mg to about 790 mg, or about 600 mg to about 790 mg, or about 650 mg to about 790 mg, or about 700 mg to about 790 mg, or about 710 mg to about 790 mg, or about 720 mg to about 790 mg, or about 730 mg to about 790 mg, or about 740 mg to about 790 mg, or about 750 mg to about 790 mg, or about 500 mg, or about 550 mg, or about 600 mg, or about 650 mg, or about 700 mg, or about 750 mg.

8. The method according to any one of claims 1-7, wherein, The anti-CD20 antibody or its antigen-binding fragment may be administered as a single dose, or once a week, month, every two months, every three months, every four months, every five months, every six months, every seven months, every eight months, every nine months, every ten months, every eleven months, or every twelve months.

9. The method according to any one of claims 1-7, wherein, The anti-CD20 antibody or its antigen-binding fragment is administered approximately every six months.

10. The method according to claim 5 or 6, wherein, The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 20 mg, administered approximately every four months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 20 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 100 mg, administered approximately every five months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 100 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 200 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 500 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 500 mg, administered approximately every five months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 500 mg, administered approximately every four months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 550 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 600 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 650 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 700 mg, administered approximately every six months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 750 mg, administered approximately every four months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 750 mg, administered approximately every five months; or The dosage of the anti-CD20 antibody or its antigen-binding fragment is approximately 750 mg, administered approximately every six months; or The anti-CD20 antibody or its antigen-binding fragment is administered as needed due to disease recurrence.

11. The use of an effective amount of anti-CD20 antibody or its antigen-binding fragment in the preparation of a medicament for treating a disease; wherein, The anti-CD20 antibody or its antigen-binding fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:1; and the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

2. The effective dose is approximately 710 mg to approximately 790 mg per dose.

12. The use according to claim 11, wherein, The diseases are those associated with CD20 expression, such as oncogenic and immune diseases; the oncogenic diseases include B-cell lymphomas, which include Hodgkin's lymphoma and non-Hodgkin's lymphoma; the immune diseases include multiple sclerosis, immune thrombocytopenic purpura, neuromyelitis optica, myasthenia gravis, rheumatoid arthritis, psoriasis, and psoriatic arthritis; or The diseases mentioned are neuromyelitis optica spectrum disorders (NMOSD), non-Hodgkin's lymphoma (NHL), multiple sclerosis (MS), immune thrombocytopenic purpura (ITP), rheumatoid arthritis (RA), Wechsler's granulomatosis (WG), microscopic polyangiitis (MPA), lupus nephritis, systemic lupus erythematosus, and chronic lymphocytic leukemia (CLL); wherein, the neuromyelitis optica spectrum disorders include AQP4-IgG negative neuromyelitis optica spectrum disorders and AQP4-IgG positive neuromyelitis optica spectrum disorders.

13. The use according to claim 11 or 12, wherein, The effective dose is approximately 750 mg per dose.

14. The use according to any one of claims 11-13, wherein, The anti-CD20 antibody or its antigen-binding fragment may be administered as a single dose, or once a week, month, every two months, every three months, every four months, every five months, every six months, every seven months, every eight months, every nine months, every ten months, every eleven months, or every twelve months.

15. The use according to any one of claims 11-13, wherein, The anti-CD20 antibody or its antigen-binding fragment is administered approximately every six months.

16. The method according to any one of claims 1-10 or the use according to any one of claims 11-15, wherein, The anti-CD20 antibody or its antigen-binding fragment is administered by injection, such as intravenous injection, subcutaneous injection, or intraperitoneal injection; or, the administration method is intravenous infusion.

17. The method according to any one of claims 1-10 or 16, or the use according to any one of claims 11-16, wherein, The light chain variable region comprises the amino acid sequence shown in SEQ ID NO:1, and the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:2; and / or The anti-CD20 antibody comprises a light chain and a heavy chain, wherein the light chain comprises an amino acid sequence as shown in SEQ ID NO:3 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:3, and the heavy chain comprises an amino acid sequence as shown in SEQ ID NO:4 or an amino acid sequence having at least 80% sequence identity with SEQ ID NO:

4.

18. The method according to any one of claims 1-10 or 16-17, or the use according to any one of claims 11-17, wherein, At least about 60% of the anti-CD20 antibodies have G0 glycans at the N-glycosylation sites in the constant region of the anti-CD20 antibody; and / or The fucose-containing antibody content of the anti-CD20 antibody does not exceed about 5%; and / or The anti-CD20 antibody was produced by a CHO cell line in which the α-(1,6) fucosyltransferase gene was knocked out.

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