Inerizumab and methods of using same for sustained b-cell depletion

Abstract

Disclosed herein are compositions and methods for treating autoimmune diseases using irnerizumab. In particular, the use of irnerizumab for the treatment of neuromyelitis optica lineage disorders and exemplary treatment regimens are provided.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 369,590, filed on July 27, 2022, which is incorporated herein by reference in its entirety for all purposes.

[0003] References to Electronic Sequence Listings

[0004] The contents of the electronic sequence listing (HOPA_045_01WO_SeqList_ST26.xml; size: 10,611 bytes; creation date: September 19, 2022) are incorporated herein by reference in its entirety. Technical Field

[0005] The present disclosure relates to compositions comprising inerizumab and methods of using the same for sustained B cell depletion. Background Art

[0006] Inelizumab is an anti-CD19 monoclonal antibody that has been approved in Japan and the United States for the treatment of neuromyelitis optica spectrum disorder (NMOSD). However, the relationship between B cell, plasma cell, and immunoglobulin depletion after inelizumab treatment and the reduction in NMOSD disease activity after inelizumab treatment remains unclear, and further elucidation of this relationship requires analysis to clarify the optimal treatment approach for subjects with NMOSD. Summary of the invention

[0007] This specification provides a method for treating or preventing neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need of B cell levels less than about 4 cells / μL of blood, thereby treating the NMOSD. In some aspects, the Fcγ receptor IIIa (FCGR3A) gene of the subject in need comprises the rs396991 polymorphism. In some aspects, the subject in need comprises a V allele determined by a V / F or V / V genotype.

[0008] Provided are methods of treatment, including analyzing a blood sample from a subject in need to determine the presence of the rs396991 polymorphism of the FCGR3A gene, and if the rs396991 polymorphism is determined to be positive, administering inelizumab to the subject in need. In some aspects, the subject in need suffers from an autoimmune disease. In some aspects, the autoimmune disease is selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG) and IgG4RD. In some aspects, the autoimmune disease is NMOSD. In some aspects, the autoimmune disease is MG. In some aspects, the autoimmune disease is IgG4RD. In some aspects, the subject in need suffers from at least two autoimmune diseases. In some aspects, at least two autoimmune diseases are selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG) and IgG4RD.

[0009] A method of treating neuromyelitis optica spectrum disorder (NMOSD) is provided, the method comprising administering inelizumab to a subject in need thereof comprising the rs396991 polymorphism of the FCGR3A gene, wherein the administration is more effective in reducing the level of B cells or plasma cells in the subject in need thereof comprising the rs396991 polymorphism of the FCGR3A gene compared to an otherwise comparable subject lacking the rs396991 polymorphism.

[0010] In some respects, the level of B cell or plasma cell is determined after at least 1,2,4,8,12,16,22,28 and / or 32 weeks of this application.In some respects, use the standard of the group of being selected from following composition effectively to reduce: annual attack rate (AAR), disability incidence rate, hospitalization incidence rate, magnetic resonance imaging (MRI) lesions and combination thereof assessed by expanded disability status scale (EDSS) score.In some respects, use the AAR effectively to reduce, wherein this is reduced to at least about 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or up to about 100%.In some respects, use these MRI lesions effectively to reduce, wherein this is reduced to at least about 20%, 30%, 40%, 50%, 60%, 70%, 73%, 75%, 80%, 85%, 90%, 95%, 97% or up to about 100%. In some aspects, the administration is repeated. In some aspects, the administration is repeated every day, every month, or every year. In some aspects, the administration is repeated every month. In some aspects, the administration is repeated every 6 months. In some aspects, the administration lasts for at least about 1 year, 2 years, 3 years, 4 years, 5 years, or for about the lifetime of the subject in need. In some aspects, the subject in need has previously administered inelizumab. In some aspects, the administration is intravenous. In some aspects, the administration is performed by intravenous infusion. In some aspects, inelizumab is administered at a dosage of about 300 mg.

[0011] A method for treating or preventing an autoimmune disease is provided, the method comprising administering 300 mg of inelizumab to a subject in need whose B cell level is less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months. In some aspects, the autoimmune disease is selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD. In some aspects, the autoimmune disease is NMOSD. In some aspects, the autoimmune disease is MG. In some aspects, the autoimmune disease is IgG4RD.

[0012] A method of treating or preventing neuromyelitis optica spectrum disorder (NMOSD) is provided, the method comprising administering 300 mg of inerizumab to a subject in need thereof having a B cell level of less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months.

[0013] Provided is a method for treating or preventing an autoimmune disease, the method comprising administering 300 mg of inelizumab to a subject in need thereof having a B cell level of less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months, and wherein the autoimmune disease is selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4-related disease (IgG4RD). In some aspects, administration continues for at least about 2.5 years.

[0014] Provided are methods for treating neuromyelitis optica spectrum disorders, including administering inelizumab to a subject in need, wherein the subject does not have anti-aquaporin 4 immunoglobulin G antibodies (AQP4-IgG-), and has immunoglobulin G antibodies (MOG-IgG+) for myelin oligodendrocyte glycoprotein. In some aspects, administering about 300 mg of inelizumab. In some aspects, administering continues for at least 6 months, 1 year, 2 years, or 3 years. In some aspects, administering continues indefinitely. In some aspects, administering continues for at least 2 years. In some aspects, compared to healthy control subjects, subjects in need have increased serum autoreactivity to autoantibodies unrelated to NMOSD. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1A Illustration designed for the study. Figure 1B The probability of being seizure-free during RCP is shown for newly diagnosed subjects. Figure 1C Shown are the probability of being seizure-free for subjects with ≥2 prior NMOSD relapses.

[0016] Figure 2A Serum autoreactivity of all subclasses, IgG subclass, IgM subclass, IgA subclass, and IgE subclass to at least one autoantigen in subjects with NMOSD is shown. Figure 2B Shown are subjects with NMOSD who were IFNGS (type 1 interferon-inducible gene signature) positive at baseline. Figure 2C The proportion of IFNGS-negative and IFNGS-positive subjects with NMOSD that were reactive to at least one anti-ribonucleoprotein (RNP) IgG is shown. Figure 2D The ratio of ANA (antinuclear antibody) scores by IFNGS status is shown. Figure 2E Shown are (left) ANA scores, (middle) autoantibody scores, and (right) IFNGS positivity for subjects with NMOSD (alone) or with other coexisting inflammatory diseases.

[0017] Figure 3A The kinetics of B cell depletion after treatment with inerizumab are shown. Figure 3BLong-term (>2.5 years) B cell depletion kinetics for each subgroup are shown. Figure 3C The AAR is displayed. Figure 3D Annualized rates of new / enlarging T2 lesions are shown. Figure 3E The annualized rate of worsening EDSS scores is shown. Figure 3F Annualized rates of hospitalization for subjects with B cell counts a≤4 cells / μL relative to subjects with B cell counts >4 cells / μL are shown. a B cell counts were measured after the first inelizumab dosing interval (6 months after treatment initiation). Figure 3G AAR and new / emerging T2 lesions are shown. Figure 3C-3G In each of the Figures, these graphs show the ratio of placebo-treated subjects (in gray) during the RCP, followed by two groups of inelizumab-treated subjects (in green and light blue) during the first 6 months of inelizumab treatment (first dosing interval) and annually thereafter. The last bar in each figure shows the combined ratio of each endpoint after 2.5 years or more of continued inelizumab treatment. Error bars represent 95% CI estimated by negative binomial regression. AAR, annualized attack rate; CI, confidence interval; EDSS, expanded disability status scale; RCP, randomized controlled period.

[0018] Figure 4A-4C To show the effect of long-term (>2.5 years) inelizumab-associated B-cell depletion on disease activity measures.

[0019] Figure 5A Schematic illustration of CD16a-mediated ADCC. Figure 5B (Left) AAR and (right) EDSS scores from subjects grouped by FCGR3A genotype. Figure 5C Shown are the risk of flare (left), rate of new / enlarging T2 lesions (center), and rate of RCP hospitalization (right) for placebo-treated subjects. Figure 5D Shown are the flare risk (left) and rate of new / enlarging T2 lesions (right) for inelizumab-treated subjects. Figure 5E Longitudinal CD20+ B cell counts (left), longitudinal plasma cell profiles (middle), and longitudinal IgG (right) by FCGR3A genotype are shown. Fig. 5F Annualized rates of episodes, expanded disability status scale, hospitalizations, and new lesions are shown.

[0020] Figure 6A-6F The kinetics of B cell and immunoglobulin depletion are demonstrated. Fig. 6A The plasma cell gene signature normalized to a pool of 10 healthy donors using the ddCT method is shown over time. Figure 6B Total immunoglobulin over time is shown. Figure 6Cshows the change of immunoglobulin G over time, Fig.6D shows the changes in immunoglobulin M over time, Fig. 6E shows the changes in immunoglobulin A over time, and Fig. 6F Immunoglobulin E over time is shown for subjects treated with placebo (grey) and inelizumab (red), during the RCP period. Points represent medians and shaded areas represent IQRs. The black dashed line represents the LLOQ of 0.2 cells / μL determined by FACS. Samples with counts below the LLOQ were estimated to be 0·05 cells / μL. *p<0·05, **p<0·01, ***p<0·001, Mann-Whitney U test between dose groups. The vertical dashed lines indicate that the subject transitioned to the OLP and two inelizumab doses were administered to the placebo group; an additional maintenance dose was administered to the inelizumab group. ddCT=ΔΔ cycle threshold. FACS=fluorescence activated cell sorting. Ig=immunoglobulin. IQR=interquartile range. LLOQ=lower limit of quantitation. OLP=open label period. RCP=randomized controlled period.

[0021] Figure 7A-7D showed the reduction in NMOSD progression observed under long-term inelizumab treatment. Fig. 7A Annualized attack rates (AAR) are shown. Figure 7B Annualized rates of new / enlarging T2 lesions are shown. Figure 7C The annualized rates of EDSS worsening are shown. Fig.7D The annualized hospitalization rates for placebo-treated subjects during the RCP period, for all subjects during the first dosing period of inelizumab treatment (first dosing interval), and each year thereafter are shown. The last column in each figure shows the combined rate of each endpoint after 2.5 years or more of continuous inelizumab treatment. Error bars represent 95% CI estimated by negative binomial regression.

[0022] Figure 8A-8C The temporal relationship between CD20+ B cell counts and onset is shown. Fig. 8A Shown is an overview of CD20+ B cell counts in subjects who experienced a flare following inelizumab treatment. Figure 8B Box plots of CD20+ B cell counts in samples drawn before a flare (samples drawn ranging from 15-115 days before a flare), during a flare assessment, and after a flare (samples drawn ranging from 8-200 days after a flare) for all samples drawn from subjects who never experienced a flare (left). Figure 8C Box plot of CD20 B cell counts in a subset of samples drawn after administration of the third dose.

[0023] Figure 9A-9BB cell kinetics during the first dosing interval of inelizumab are shown, predicting long-term profound B cell depletion. Fig. 9A Figure 3 is a profile of B cell counts for individual subjects during the first dosing period of inelizumab. Fig. 9B Total CD20 counts during 3 years of inelizumab treatment in subjects with B cell counts >4 and ≤4 cells / μL at the end of the first dosing period are shown. Points represent median + / - IQR. (*p<0·05, **p<0·01, ***p<0·001, Mann-Whitney U test). IQR = interquartile range. RCP = randomized controlled period.

[0024] Fig.10 Annualized hospitalization rates by subjects with >4 cells / μl after the first inelizumab dosing interval (6 months after treatment) are shown.

[0025] Fig.11 is an exemplary diagram showing that CD19 is a differentiation target for B cell depletion.

[0026] Figure 12A-12B B cell counts for individual subjects at the time of flare are shown. For subjects who experienced a flare with more than 2 doses of inerizumab, Fig. 12A Total CD20+ cell counts over time are shown, and Fig. 12B CD20+ memory B cell counts over time are shown. Each sub-panel contains data for five subjects; each subject is represented by a different colored line.

[0027] Figures 13A-13D Figure 2 is a graph showing the rate ratio of disease activity based on the B cell level cutoff after the first dosing interval. The rate ratio (+ / -95% CI shaded area) is relative to the B cell cutoff for W26 onset that occurred after the first dosing period with inelizumab ( Fig.13A ), new or enlarging T2 lesions ( Fig. 13B ), EDSS worsening (subject / year) ( Fig. 13C ), hospitalization( Fig.13D ). Rate ratios were calculated by comparing the rate of each endpoint occurring in subjects below each B-cell cutoff relative to those above each B-cell cutoff. AAR = Annualized Attack Rate. CI = Confidence Interval. EDSS = Expanded Disability Status Scale. RR = Rate Ratio.

[0028] Fig.14A Annualized attack rates are shown. Fig. 14BThe annualized rates of new / enlarging T2 lesions are shown. After the first inelizumab dosing interval (6 months after treatment) in AQP4-seropositive subjects, divided by subjects with counts above or below 4 cells / μL, Fig. 14C The annualized rate of EDSS worsening is shown and Fig.14D Annualized rates of hospitalizations are shown.

[0029] Fig.15 The pharmacokinetics of inelizumab in subjects with B cell counts above or below 4 cells / μl at 6 months are shown. During the first dosing period of inelizumab, median serum inelizumab concentrations in subjects with B cells ≤ 4 cells / μL (blue), and subjects with B cells ≤ 4 cells / μL (blue) or subjects with B cells> 4 cells / μL (red) (upper graph). During Week 22 of RCP, serum inelizumab concentrations in both groups (lower graph).

[0030] Figure 16A-16C Baseline B cell counts, plasma cell gene signatures, and total Ig are shown for subjects stratified by B cell counts after the first dosing period of inerizumab. Fig.16A Baseline blood CD19+ B cell counts are shown. For the two B cell depletion subgroups, Fig. 16B The plasma cell gene signature was revealed. Fig. 16C Total immunoglobulins are shown. FC = fold change. HD = healthy donor. Ig = immunoglobulin.

[0031] Fig.17A Median B cell counts are shown for subjects with B cell counts ≤ 4 cells / ul after the first dosing period of inelizumab treatment (blue), and for subjects with B cell counts > 4 cells / ul at the end of the first dosing period (these subjects were further subdivided by whether the subject showed depletion of ≤ 4 cells / ul at any time during this interval (red and blue)). Fig. 17B Bar graph showing the % of subjects homozygous for rs396991F / F within each subgroup.

[0032] Figures 18A-18C The relationship between CD20+ B cell count and ADA is shown. Fig.18A Figure 2 is a profile curve of ADA titer changes over time since the start of inelizumab treatment. Fig.18B Figure 2 is a graph showing the percentage of subjects with B cell counts > 4 cells / μL in different ADA subgroups at W26-W28. Fig. 18CBox plots of median B cell counts for subjects who exhibited persistent ADA negativity, subjects who exhibited intermittent ADA positivity at intervals >90 days from ADA positivity, and for these subjects at intervals within 90 days of ADA positivity. ADA = anti-drug antibodies. BP = baseline positive. LLN = lower limit of normal. LLOQ = lower limit of quantitation. TU = treatment unaffected. TE = emerging after treatment. W = week.

[0033] Fig.19A An exemplary study design is shown. Fig.19B The time from the first dose of inerizumab to the onset determined by the first adjudication committee (AC) is shown. Fig.19C CD19 B cell levels of seven subjects with serotype AQP4-IgG- / MOG-IgG+ are shown. DETAILED DESCRIPTION

[0034] Described herein is inelizumab and methods of using it to reduce B cell-mediated immune responses in subjects with a disease or disorder. Also provided are methods including subject / subject stratification to optimize inelizumab treatment of NMOSD based on the presence of biomarkers (such as polymorphisms of the FCGR3A gene). Also provided are treatment regimens comprising inelizumab for the treatment of NMOSD.

[0035] definition

[0036] Although it is believed that the following terms are well understood by those of ordinary skill in the art, the following definitions are provided to facilitate the interpretation of the subject matter of this disclosure. Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those generally understood by those of ordinary skill in the art. References to the technology employed herein are intended to refer to technology as generally understood in the art, including variations of those technologies that are apparent to those of ordinary skill in the art and / or replacement of equivalent technologies.

[0037] Any range listed herein includes the endpoints. For example, a range of 2-4 includes 2 and 4 and values ​​therebetween.

[0038] As used herein, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0039] When immediately preceding a numerical value, the term "about" or "approximately" means a range of plus or minus 10% of the value. For example, unless the context of the present disclosure otherwise indicates or is inconsistent with such an interpretation, "about 50" may mean 45 to 55, "about 25,000" may mean 22,500 to 27,500, etc. In the context of a numerical list such as "about 49, about 50, about 55, ...", "about 50" means a range extending to less than half of the interval between the preceding and following values, for example, greater than 49.5 to less than 52.5. In addition, the phrase "less than about" a value or "greater than about" a value should be understood according to the definition of the term "about" provided herein.

[0040] The term "identity" when referring to nucleic acid sequences or protein sequences is used to indicate the similarity between two sequences. Unless otherwise stated, percent identity described herein is determined using the BLAST algorithm available on the World Wide Web at: blast.ncbi.nlm.nih.gov / Blast.cgi using default parameters.

[0041] As used herein, the term "subject" refers to any subject, such as a human or non-human mammal, that needs diagnosis, prognosis or therapy. The term "subject" may mean a person or non-human mammal that suffers from, may suffer from or is suspected of having a disease (such as an autoimmune disease or illness). The terms "subject" and "patient" are used interchangeably herein. Although the compositions provided herein are mainly related to compositions suitable for use to people, it will be understood by those skilled in the art that such compositions are generally suitable for use to subjects of all classifications. In some aspects, the subject is a mammal. Mammals include primates, such as humans, monkeys, chimpanzees and apes, and non-primates, such as domestic animals, including laboratory animals (such as rabbits and rodents, such as guinea pigs, rats or mice) and domestic pets and farm animals (such as cats, dogs, pigs, cattle, sheep, goats, horses, rabbits), and non-domestic animals, such as wild animals, birds, reptiles, fish, etc.

[0042] As used herein, "treating" or "treat" describes the management and care of a subject for the purpose of combating a disease, condition, or disorder, and includes administering the composition used in the methods described herein to alleviate the symptoms or complications of the disease, condition, or disorder, or to eliminate the disease, condition, or disorder. Therefore, the term "treat / treating" refers to therapeutic measures and preventive or prophylactic measures, wherein the goal is to prevent, slow down (mitigate), or improve the progression of a disease (e.g., an autoimmune disease). Beneficial or desired clinical results include, but are not limited to, symptom relief, reduced disease severity, stable disease state (i.e., no worsening), delayed or slowed disease progression, improved or mitigated disease state, and disease reversal (whether partial or complete). The term "treatment" may also include treatment of in vitro cells or animal models.

[0043] Inerizumab

[0044] Inelizumab (also known as MEDI-551) is a humanized, affinity-optimized, afucosylated IgG1κ monoclonal antibody (mAb), called 16C-aFuc, that binds to the B-cell-specific surface antigen CD19, thereby reducing or eliminating CD19+ B cells. Inelizumab was glycoengineered by expressing mAb 16C4 in a Chinese hamster ovary producer cell line that lacks fucosyltransferase (BioWa Technology) to produce a homogeneous afucosylated antibody with enhanced antibody-dependent cellular cytotoxicity. In contrast to the anti-CD20 mAb rituximab, inelizumab does not mediate complement-dependent cytotoxicity, but eliminates B cells via antibody-dependent cellular cytotoxicity and antibody-mediated cellular phagocytosis mechanisms.

[0045] In some aspects, inelizumab comprises a CDR sequence from Table 1. In some aspects, inelizumab comprises a variable heavy chain CDR1-CDR3 corresponding to SEQ ID NOs: 2-4. In some aspects, inelizumab comprises a variable light chain CDR1-CDR3 corresponding to SEQ ID NOs: 6-8. In some aspects, inelizumab comprises a VH amino acid sequence and a VL amino acid sequence of Table 1 corresponding to SEQ ID NOs: 1 and 5, respectively. In some aspects, inelizumab comprises a heavy chain amino acid sequence and a light chain amino acid sequence of Table 1 corresponding to SEQ ID NOs: 9 and 10, respectively.

[0046] In some aspects, inelizumab or a derivative of inelizumab has at least about or up to about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or up to about 100% identity with any of SEQ ID NO: 1-SEQ ID NO: 10 shown in Table 1. In some aspects, any of the sequences from Table 1 can be modified. In some aspects, the modification comprises one or more truncations, deletions, insertions, and combinations thereof. In some aspects, the modification does not alter the function of inelizumab. The modification may be present at any of the residues provided in Table 1 and at any number of residues from Table 1. In some aspects, the modification may comprise 1-3, 1-5, 1-10, 5-20, 1-3, 1-5, 1-10, 1-20, 3-8, 3-10, 3-15, 5-8, 5-10, or 5-20 residues. In some aspects, the modification may be present in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, or 450 residues. In some aspects, all residues of Table 1 comprise modifications.

[0047] Table 1: Exemplary Inerizumab Sequences

[0048]

[0049] Inerizumab and methods for its preparation are described in International PCT Patent Application PCT / US2007 / 077916 (published as WO 2008 / 031056), which is hereby incorporated by reference (PCT / US2007 / 077916 refers to VIB551 as "16C4"). In some aspects, VIB551 (also known as HZN551, MEDI551, UPLIZNA TM or inelizumab; disclosed in U.S. Application No. 11 / 852,106 and International Application No. PCT / US2020 / 029613, which are incorporated herein by reference in their entireties) administered in any of the methods disclosed herein.

[0050] In some aspects, inelizumab is transparent to light milky white, colorless to light yellow solution, does not contain or does not actually contain visible particles. Inelizumab is a sterile liquid medicine (100mg inelizumab per vial, nominal) for IV infusion after dilution in normal saline. In some aspects, inelizumab for IV administration can be provided as a sterile liquid in a 10R glass vial with a nominal fill volume of 10mL with a 20mm stopper and a flip-top upper seal. In some aspects, the formulation can be preservative-free. In some aspects, inelizumab for IV administration is provided in the form of a sterile liquid filled in a 10R vial with a nominal volume of 10mL. In some aspects, the vial contains 10 mg / mL of inelizumab formulated in 20 mM histidine / histidine hydrochloride, 70 mM NaCl, 106 mM (4% [w / v]) trehalose dihydrate, and 0.01% (w / v) polysorbate 80, pH 6.0. Alternative formulations are also contemplated and described further below.

[0051] method

[0052] In some aspects, inelizumab is used to treat autoimmune diseases or disorders. In some aspects, the present disclosure provides a method for treating neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need of treating NMOSD. In some aspects, inelizumab can be administered as a first-line therapy.

[0053] In some aspects, the autoimmune disease is selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG) and IgG4RD. In some aspects, the autoimmune disease or disorder comprises neuromyelitis optica spectrum disorder (NMOSD). In some aspects, the autoimmune disease or disorder comprises MG. In some aspects, the autoimmune disease or disorder comprises IgG4RD.

[0054] The treatment of autoimmune disease or disorder can be in the form of suppressing B cell or T cell mediated immune response, which can be the reduction of class switching antibodies, the reduction of circulating B cell subsets, the reduction of plasma activity or the reduction of plasma cells and plasma cell gene characteristics. The treatment of autoimmune disease or disorder can be the reduction of inflammatory markers. Inflammatory markers can be one or more of the following: autoantibody levels, plasma cells (PC) or PC gene characteristics (characterized by the expression of genes IGHA1, IGJ, IGKC, IGKV4-1 and TNFRSF17), circulating B cell subsets and class switching antibodies. The treatment of autoimmune disease or disorder can be the reduction of clinical signs and symptoms, such as clinical signs and symptoms measured by subject or physician overall assessment. In some aspects, one or more of the following are effectively reduced by administration: annual attack rate (AAR), disability incidence assessed by expanded disability status scale (EDSS) score, hospitalization incidence, magnetic resonance imaging (MRI) lesions and combinations thereof.

[0055] Treatment of an autoimmune disease or disorder may be characterized by a reduction in clinical symptoms of the disease or disorder by at least 20%, or a reduction in inflammation, or a reduction in a biomarker of the disease or disorder relative to its level prior to treatment with inelizumab. The reduction in any of these symptoms or inflammation or biomarkers may be a reduction in symptoms or inflammation or biomarkers by at least 50% relative to its level prior to starting treatment with inelizumab. This reduction may characterize the autoimmune disease or disorder as being in remission.

[0056] In some aspects, the autoimmune disease or disorder is NMOSD. NMOSD is a rare autoimmune disease of the central nervous system (CNS), in which approximately 75%-90% of subjects have pathogenic immunoglobulin (Ig) G autoantibodies (AQP4-IgG) against aquaporin-4 water channels. Repeated severe attacks of optic neuritis and myelitis are typical manifestations of the disease, but brain and brainstem lesions may also occur. B cells play a role in the pathogenesis of NMOSD through a variety of mechanisms. Exemplary mechanisms include: B cell secretion of proinflammatory cytokines (such as interleukin (IL) -6 and IL-17), which may stimulate pathogenic, proinflammatory immune responses. Secondly, B cells can act as antigen presenting cells to promote the development and activation of autoimmune T cells. Third, B cell maturation leads to the generation of plasmablasts and plasma cells that produce AQP4-IgG.

[0057] If inelizumab is used to treat NMOSD, it may treat NMOSD by reducing the worsening of the subject's Kurtzke Expanded Disability Severity Scale (EDSS), or reducing the number of active magnetic resonance imaging (MRI) lesions in the subject, or reducing the worsening of the subject's modified Rankin score, or reducing the frequency of the subject's NMOSD-related hospitalizations, or reducing the subject's risk of NMOSD-related attacks, or reducing optic neuritis, or reducing the severity of the subject's NMOSD-related attacks, or reducing the subject's pain, or reducing the subject's NMOSD-related impairment, or reducing the subject's NMOSD-related attacks.

[0058] If inelizumab treats the subject's NMOSD by reducing the deterioration of the subject's EDSS score, and the subject's baseline EDSS score is 0, the subject's EDSS score may deteriorate by less than 2 points, or deteriorate by less than 1 point, or deteriorate by less than 0.5 points. For subjects with a baseline score of 0, this reduction in deterioration of the EDSS score may last for a period of at least 6 months, 9 months, 1 year, 2 years, 3 years, 4 years, 5 years, 7.5 years, or 10 years. If inelizumab treats the subject's NMOSD by reducing the deterioration of the subject's EDSS score, and the subject's baseline score is 1 to 5, the subject's EDSS score may deteriorate by less than 1 point, or deteriorate by less than 0.5 points. For subjects with a baseline EDSS score of 1 to 5, this reduction in deterioration may be a reduction in deterioration that lasts for a period of more than 6 months, 9 months, 1 year, 2 years, 3 years, 4 years, 5 years, 7.5 years, or 10 years. If inelizumab treats the subject's NMOSD by reducing the worsening of the subject's EDSS score, and the subject's baseline EDSS score is 5.5 or higher, the subject's EDSS score may worsen by less than 0.5 points, or worsen by less than 0.25 points. For subjects with a baseline score of 5.5 or higher, this reduction in worsening may be a reduction in worsening of the EDSS score that lasts for a period of more than 6 months, 9 months, 1 year, 2 years, 3 years, 4 years, 5 years, 7.5 years, or 10 years.

[0059] If inelizumab treats NMOSD in a subject by reducing the number of active MRI lesions, the treatment may be a reduction in the number of enlarged T2 MRI lesions, gadolinium-enhanced (Gd+) lesions, or may be a reduction in the number of new MRI lesions, or may be a reduction in the number of enlarged T2 MRI lesions, gadolinium-enhanced (Gd+) lesions, and the number of new MRI lesions. The lesion reduction may be a reduction in brain lesions, a reduction in brainstem lesions, a reduction in spinal cord lesions, a reduction in optic nerve lesions, or a reduction in lesions in any combination of two or more of the brain, brainstem, spinal cord, and optic nerves. The new MRI lesions may be asymptomatic.

[0060] If inelizumab treats NMOSD in a subject by reducing the risk of an NMOSD-related attack, the subject's risk of an attack may be reduced by between 60% and 85%, or may be reduced by between 65% and 75%, or may be reduced by between 70% and 80%. The subject's risk of an attack may be reduced by at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, or at least 80%. The subject's risk of an attack may be reduced by 70%, 75%, 76%, 77%, 78%, 79%, or 80%.

[0061] If inelizumab treats NMOSD in a subject by reducing the risk of an NMOSD-related episode, the probability of the treated subject being free of an NMOSD-related episode may be greater than 70% for at least 6 months after treatment, or greater than 70% for at least 12 months after treatment, or greater than 70% for at least 18 months after treatment due to the reduced risk of an NMOSD-related episode. The probability of the treated subject being free of an NMOSD-related episode may be greater than 75% for at least 6 months after treatment, or greater than 75% for at least 12 months after treatment, or greater than 75% for at least 18 months after treatment due to the reduced risk of an NMOSD-related episode. Additionally, the probability of the treated subject being free of an NMOSD-related episode may be greater than 80% for at least 6 months after treatment, or greater than 80% for at least 12 months after treatment, or greater than 80% for at least 18 months after treatment due to the reduced risk of an NMOSD-related episode. Furthermore, due to the reduced risk of NMOSD-related episodes, the probability of a treated subject being free of an NMOSD-related episode can be greater than 85% for at least 6 months after treatment, or greater than 85% for at least 12 months after treatment, or greater than 85% for at least 18 months after treatment.

[0062] In addition, if inelizumab treats the subject's NMOSD by reducing the risk of an NMOSD-related episode, the annual risk of an NMOSD-related episode for the treated subject may be reduced to between 0.18 and 0.08 due to the risk reduction, or it may be reduced to between 0.15 and 0.08, or it may be reduced to 0.14, or 0.13, or 0.12, or 0.11, or 0.10, or 0.09, or 0.08 or 0.07. If the subject being treated for NMOSD is AQP4-IgG seropositive, the subject's annual risk of an NMOSD-related episode may be reduced to between 0.15 and 0.11, or to between 0.14 and 0.12, or it may be reduced to 0.14, 0.13, 0.12, or 0.11. If the subject being treated for NMOSD is AQP4-IgG seronegative, the subject's annual risk of NMOSD-related episodes may be reduced to between 0.09 and 0.07, or it may be reduced to 0.09, 0.08, or 0.07. NMOSD-related episodes (whose risk may be reduced with treatment of NMOSD subjects) may be episodes characterized by the appearance of new NMOSD symptoms or worsening of existing NMOSD symptoms. The new or existing symptoms may be ocular symptoms. If the new or existing symptoms are ocular symptoms, they may be eye pain, new optic neuropathy, enlarged optic neuropathy, blurred vision, loss of vision, or a decrease of 5 or more characters on the low-contrast Landolt C Broken Rings Chart. The new or existing symptoms may be spinal symptoms. If the new or existing symptoms are spinal symptoms, they may be deep or radicular pain, paresthesias in the limbs, weakness, sphincter dysfunction, Lhermitte's sign, new spinal cord lesions, or enlarged spinal cord lesions. The new or existing symptom may be a brain or brainstem symptom. If the new or existing symptom is a brain or brainstem symptom, it may be nausea, diplopia, oculomotor nerve palsy, vertigo, intractable vomiting, intractable hiccups, dysarthria, dysphagia, weakness, encephalopathy, hypothalamic dysfunction, new brain or brainstem lesions, or enlarged brain or brainstem lesions. The new or worsening symptom may be a combination of any two or more of the eye, spinal cord, or brain / brainstem symptoms. It may be a combination of any two, three, or four of these symptoms.

[0063] In some aspects, inelizumab treats NMOSD in a subject by reducing optic neuritis, and the subject may then experience reduced eye pain, reduced vision loss, reduced visual field loss, reduced color vision loss, or a reduction in flashing or flickering lights that move with the eye. Reduction in optic neuritis may result in improved vision and / or relief of eye pain.

[0064] In some aspects, inelizumab treats the NMOSD of the subject by alleviating the severity of the NMOSD-related episodes of the subject, and then the severity of any NMOSD-related episodes suffered by the subject can be rated as mild or moderate, rather than being rated as severe. Mild episodes can be short-term episodes, can be episodes that only require minimal treatment or therapeutic intervention, and / or are episodes that may not interfere with common daily life activities. Moderate episodes can be episodes that can be alleviated with specific additional therapeutic interventions. Any moderate episode can be an episode that interferes with common daily life activities and / or causes discomfort but does not cause significant or permanent risk of injury to the subject. The reduction of the severity of the NMOSD-related episodes of the subject can be the reduction of the episodes that are rated as severe suffered by the subject. Such severe episodes can be episodes that require intensive therapeutic interventions, interrupt common daily life activities, or significantly affect the clinical status of the subject. Such severe episodes may require hospitalization.

[0065] In some aspects, the method includes subject stratification. Subject stratification can be based on the B cell level of less than about 4 cells / μL blood of the subject after the first or previous administration of inelizumab. In some aspects, the method includes repeated administration of inelizumab to subjects with B cell counts less than 4 cells / μL after the first dosing period. In some aspects, the method includes maintaining B cell suppression or reduction in subjects in need.

[0066] Inelizumab can also be used in a method for reducing the AQP4-IgG titer of an AQP4-IgG+ subject in need of treatment for NMOSD. The present invention provides a method comprising administering inelizumab to a subject with a high AQP4-IgG titer. In some aspects, the titer is greater than about: 1:20,480. In some aspects, the present disclosure provides a method for reducing the AQP4-IgG titer of an AQP4-IgG+ subject in need of treatment for NMOSD, the method comprising administering the anti-CD19 antibody inelizumab to a subject in need of treatment for NMOSD, wherein the subject has previously been treated with an anti-CD20 antibody, wherein the subject suffers from an NMOSD attack while being treated with the anti-CD20 antibody; and wherein inelizumab is administered intravenously at a dose of 300 mg every 6 months. In some aspects, the present disclosure provides a method of reducing the titer of AQP4-IgG in an AQP4-IgG+ subject in need of treatment for NMOSD, the method comprising: administering the anti-CD19 antibody inelizumab to a subject in need of treatment for NMOSD, wherein the subject has previously been treated with an anti-CD20 antibody, wherein the subject has an NMOSD attack within 6 months of the last dose of the anti-CD20 antibody; and wherein inelizumab is administered intravenously at a dose of 300 mg every 6 months. In some aspects, administration can continue indefinitely. If inelizumab is used in a method for reducing AQP4-IgG titer in an AQP4-IgG+ subject in need of treatment for NMOSD, inelizumab may reduce AQP4-IgG titer by 75% to 100%, or 75% to 90%, or 75% to 85%, or 80% to 100%, or 85% to 100%, or 90% to 95%, or 75%, 80%, 85%, 90%, 95%, or 100%. Inelizumab may reduce AQP4-IgG titer for a duration of at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months after administration of an inelizumab dose. In some aspects, methods of treating neuromyelitis optica spectrum disorders are provided. In some aspects, these methods include administering inelizumab to a subject without anti-aquaporin 4 immunoglobulin G antibodies (AQP4-IgG-) and with immunoglobulin G antibodies (MOG-IgG+) for myelin oligodendrocyte glycoprotein. In some aspects, administering about 300mg of inelizumab. In some aspects, administering continues at least about 6 months, 1 year, 2 years, 3 years or indefinitely. In some aspects, administering continues at least 2 years. In some aspects, compared with healthy control subjects, there is a need for subjects to have increased serum autoreactivity to autoantibodies unrelated to NMOSD. In some aspects, administering effectively reduces AAR.An attack includes the presence of one or more new symptoms associated with NMOSD or a worsening of one or more existing symptoms that meets at least one of the 18 protocol-defined neurological examination criteria for an attack (including myelitis, optic neuritis, and brain / brainstem regional symptoms). In some aspects, the reduction in AAR is at least about 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or up to about 100%.

[0067] In some aspects, administration is effective to reduce MRI lesions. In some aspects, the reduction in MRI lesions is at least about 20%, 30%, 40%, 50%, 60%, 70%, 73%, 75%, 80%, 85%, 90%, 95%, 97% or up to about 100%.

[0068] In some aspects, the administration of inelizumab effectively prevents disease progression. For example, a single or repeated dosing regimen of inelizumab can effectively postpone NMOSD progression by at least about 1 month, 2 months, 3 months, 5 months, 6 months, 8 months, 10 months, 12 months, 1.5 years, 2 years, 3 years, 5 years or more than 5 years. In some aspects, the NMOSD progression of the subject administered inelizumab is reduced, as determined by the annualized rate of reduced AAR, new / enlarged T2 lesion detection or gadolinium enhanced (Gd+) lesion detection, hospitalization rate and / or EDSS deterioration.

[0069] In some aspects, the method includes determining the B cell level in peripheral blood. In some aspects, the method includes B cell monitoring. Any subject sample can be monitored, and the sample includes, but is not limited to, whole blood, PBMC, bone marrow, tissue, tumor, and combinations thereof. In some aspects, whole blood is monitored for B cells. Monitoring can be performed once or repeatedly. In some aspects, B cell monitoring is performed every day, every week, every month, every half a year, or every year. In some aspects, the level of B cells or plasma cells is determined at least 1, 2, 4, 8, 12, 16, 22, 28, and / or 32 weeks after administering inelizumab.

[0070] B cell levels can be determined using flow cytometry and / or FACS analysis. In some aspects, CD20 is used as a FACS marker (the combined inelizumab interferes with the FACS based on CD19) to count B cells. In some aspects, B cell subsets are also measured by CD20+B cells defined as CD45hi[CD3-, CD56-, CD14-], CD33-, CD20. In addition, plasmablast / plasma cell counting (CD45hi[CD3-, CD14-, CD56-], CD27+, HLA-DR hi / 低In some aspects, the level of B cell subsets in peripheral blood is determined.

[0071] In some aspects, the treated subjects comprise reduced levels of CD20+B cells, CD27+ memory B cells, or both compared to otherwise comparable subjects not treated with inelizumab. In some aspects, the treated subjects comprise at least about 1-fold, 3-fold, 5-fold, 10-fold, 20-fold, 40-fold, 60-fold, 80-fold, 100-fold, 130-fold, 150-fold, 175-fold, 200-fold, 300-fold, 400-fold, or up to about 500-fold reduction in CD27+ and CD20+B cell levels.

[0072] In some aspects, the method includes maintaining a reduced level of B cells in a subject in need thereof. In some aspects, the reduced level of B cells is maintained for at least about 1 month, 3 months, 5 months, 6 months, 8 months, 10 months, 1 year, 1.5 years, 2 years, or up to about 3 years after treatment with inerizumab.

[0073] In some aspects, method comprises determining plasma cell specific gene expression.Gene expression can be determined using the quantitative reverse transcription PCR of blood RNA.Plasma cell gene signature can be based on the expression analysis of the gene (IGHA1, IGJ, IGKV4-1 and TNFRSF17) mainly expressed by the plasma cells in the blood.Feature can be calculated as the average expression of four plasma cell specific genes on the time point of interest minus the average expression of five control genes (B2M, GAPDH, TFRC, GUSB and UBC).The multiple change of the plasma cell gene expression signature at each time point can be calculated relative to a group of healthy donor samples and interpreted as the plasma cell abundance relative to the expected prevalence rate under the non-activated immune state.

[0074] In some aspects, single nucleotide polymorphism analysis is performed on the subject sample. TaqMan single nucleotide polymorphism spectrum analysis can be used to study the possible association between the rs396991 polymorphism of the FCGR3A gene and the impaired B cell exhaustion. In some aspects, the analysis includes determining the presence of a single nucleotide substitution (SNP) from G to T at the cDNA nucleotide position 559 of the FCGR3A gene. This SNP can produce two different FcγRIIIa allotypes: at amino acid position 158, one has valine (V) and one has phenylalanine (F), referred to as FcγRIIIA-V158F polymorphism (rs396991). The rs396991 polymorphism encodes the valine / phenylalanine substitution at position 158 of FCGR3A, which is associated with reduced binding affinity to Ig Fc, and therefore may impair the ADCC mechanism optimized by the Fc region of inelizumab. In some aspects, the method includes polymorphism analysis to determine whether the subject is suitable for administration with inelizumab.

[0075] In some aspects, the method includes measuring the presence of V alleles in the subject. V alleles can be determined by V / F or V / V genotypes. The presence of V alleles can guide the administration of inelizumab. In some aspects, the method includes stratifying the subject population based on the presence of V alleles so as to be administered with inelizumab. In some aspects, the method includes administering inelizumab to a subject in need of rs396991 polymorphisms comprising FCGR3A genes. In some aspects, treatment with inelizumab can effectively reduce the B cell or plasma cell levels of subjects in need of rs396991 polymorphisms comprising FCGR3A genes (compared to subjects comparable to other aspects of the lack of rs396991 polymorphisms).

[0076] Also provided are methods including determining autoreactivity to nuclear antigens. Antinuclear antibody (ANA) tests can determine the presence of an autoimmune disease in a subject or be used to monitor the state of an autoimmune disease in a subject. Antibodies to extractable nuclear antigens (ENAs) are ribonucleoproteins (e.g., protein antigens that do not contain DNA) that can be extracted from the nucleus. Exemplary ENAs include Ro (SS-A), La (SS-B), Sm, and RNP. In some aspects, the methods include determining the level of autoreactivity in a subject before treatment and monitoring the level during treatment. In some aspects, autoreactivity is determined weekly, monthly, twice a year, or annually.

[0077] Also provided are methods comprising determining the level or presence of an autoantigen in a subject. In some respects, one or more responses (e.g., ANA and / or RNP) to an autoantigen in IgG, IgM, IgA, and IgE are assessed. In some respects, an autoantigen is relevant to the presence of an autoimmune disease. Exemplary autoantigens include: Mi-2 (CHD4), signal recognition particle (SRP), transcription intermediary factor 1-γ (TIF1-γ; TRIM 33), nuclear matrix protein-2 (NXP2; MORC3), 3-hydroxy-3-methylglutaryl-coA reductase (HMGCR), melanoma-associated differentiation gene-5 (MDA5), small ubiquitin-like modifier activating enzymes SAE-1 and SAE-2, topoisomerase-1, centromere proteins A, B, and C ("CENP"), fibrillarin, components of the small nucleolar U3 (snoRNP) complex, nucleolar phosphoprotein (NPM; B23), RNA polymerases I, II, and III, the multicomponent PMSCL; the RNA exosome complex composed of 10 proteins (most commonly targeting EXOSC9 and EXOSC10), the Ku 70 / 80 component of the DNA-dependent protein kinase, Ro52 (TRIM 21), Ro60, La (SS-B), interferon-γ-induced protein-16 (IFI16), proteinase-3 (PR3), myeloperoxidase (MPO), ACPA (anti-citrullinated protein antibody), peptidyl arginine-deiminase-4 (PAD4), ds DNA, components of Sm splicing ribonucleoprotein (subunit AG, most commonly targeted to B, B', and D), U1-RNP, ribosomal protein P, cardiolipin, anionic phospholipid / protein complexes, N-methyl-D-aspartate (NMDA) receptor, CENP-A, CENP-B, core histones, DNA polymerase β (POLB), dsDNA, EBNA1, genomic DNA, histone H1, histone H2A, histone H2B, histone H3, histone H4, Jo-1, KU (P70 / P80), La / SSB, Mi-2, PCNA, PL-7, PL-12, PM / Scl-75, PM / Scl 100, ribophosphoprotein P0, ribophosphoprotein P1, ribophosphoprotein P2, Ro / SSA (52Kda), Ro / SSA (60Kda), S100, Scl-70 / topoisomerase I, Sm, Sm / RNP, SmD, SmD1, SmD2, SRP54, ssDNA, ssRNA, U1-snRNP 68 / 70, U1-snRNP A, U1-snRNP B / B, U1-snRNP C, total histone (Histone-total), total histone citrullinated human genomic DNA, KU (P70 / P80), Nup 62, Ro / SSA (52+60), U1-snRNP, etc.In some aspects, the autoantigen is selected from the group consisting of: CENP-A, CENP-B, core histones, DNA polymerase β (POLB), dsDNA, EBNA1, genomic DNA, histone H1, histone H2A, histone H2B, histone H3, histone H4, Jo-1, KU (P70 / P80), La / SSB, Mi-2, PCNA, PL-7, PL-12, PM / Scl-75, PM / Scl100, ribophosphoprotein P0, ribophosphoprotein P1, ribophosphoprotein P2, Ro / SSA (52Kda), Ro / SSA (60Kda), S100, Scl-70 / topoisomerase I, Sm, Sm / RNP, SmD, SmD1, SmD2, SRP54, ssDNA, ssRNA, U1-snRNP 68 / 70, U1-snRNP A, U1-snRNP B / B, U1-snRNP C, total histone (Histone-total), total histone citrullinated human genomic DNA, KU (P70 / P80), Nup 62, Ro / SSA (52+60) and U1-snRNP. In some aspects, administration of inelizumab is effective to reduce the level of autoantigens in treated subjects, compared to the baseline level of the same subject or the level of comparable untreated control subjects. The level can be reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 75%, 100%, 150%, 200%, 300%, 350% or up to about 400%.

[0078] Also provided is a method including determining the level of interferon inducible gene expression. In some aspects, gene expression profiling (e.g., using microarray) can be performed to determine the level of mRNA transcripts in the biological sample of the subject. In some aspects, gene expression profiling assesses type I interferon-α / β (IFNα / β) inducible genes. The upregulation of these genes can be related to disease activity, wherein downregulation occurs when the disease is controlled with treatment (e.g., inelizumab). In some aspects, upregulation or downregulation is about 5%, 10%, 20%, 30%, 40%, 50%, 75%, 100%, 150%, 200%, 300%, 350% or up to about 400%. In some aspects, inelizumab is used to effectively reduce the level of interferon inducible gene expression for treating a subject, compared with the baseline level of the same subject or the level of a comparable untreated control subject.

[0079] Pharmaceutical composition

[0080] Pharmaceutical compositions are provided. Also provided are pharmaceutical compositions comprising inelizumab. In some aspects, a treatment regimen comprises a pharmaceutical composition, such as a pharmaceutical composition comprising inelizumab. A treatment regimen may also comprise an additional pharmaceutical composition that is not inelizumab.

[0081] Inelizumab can be administered at any dose. In some aspects, inelizumab is administered at a dose of about 0.5 mg to 1000 mg. In some aspects, inelizumab is administered at a dose of about 10 mg to 30 mg, 20 mg to 100 mg, 50 mg to 200 mg, 100 mg to 300 mg, 150 mg to 350 mg, 200 mg to 400 mg, 250 mg to 450 mg, 300 mg to 500 mg, 300 mg to 700 mg, or 400 mg to 1000 mg.In some aspects, inelizumab is administered at a dose of about 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg g, 235mg, 240mg, 245mg, 250mg, 255mg, 260mg, 265mg, 270mg, 275mg, 280mg, 285mg, 290mg, 295mg, 300mg, 305mg, 310mg, 315mg, 320mg, 325mg, 330mg, 335mg, 340mg, 345mg, 350mg, 355mg, 360mg, 365mg, 370mg, 375mg, 380mg, 385mg, 390mg, 395mg, 400mg, 405mg, 410mg, 415mg, 420mg, 42 5mg, 430mg, 435mg, 440mg, 445mg, 450mg, 455mg, 460mg, 465mg, 470mg, 475mg, 480mg, 485mg, 490mg, 495mg, 500mg, 505mg, 510mg, 515mg, 520m g, 525mg, 530mg, 535mg, 540mg, 545mg, 550mg, 555mg, 560mg, 565mg, 570mg, 575mg, 580mg, 585mg, 590mg, 595mg, 600mg, 605mg, 610mg, 615mg, In some aspects, 300 mg is administered to a subject in need thereof.

[0082] In some aspects, inelizumab is administered repeatedly. Administration may be daily, monthly, or annual. Inelizumab dosages may be administered approximately every other week or may be administered twice a month. Inelizumab dosages may also be administered approximately weekly or approximately once a month. Inelizumab dosages may be administered every 7 days, every 10 days, every 14 days, every 15 days, every 16 days, every 14-10 days, every 14-16 days, or every 30 days. In some aspects, inelizumab is administered every 6 months. Inelizumab may be administered for at least about 1 year, 2 years, 2.5 years, 3 years, 4 years, 5 years, or for the lifetime of a subject in need. In some aspects, administration continues for at least about 2.5 years.

[0083] If the dose of inelizumab administered is 1000 mg, 1500 mg, or one between about 1000 mg and about 1500 mg, the dose may be administered every other week, or may be administered twice a month. If the inelizumab dose is 3000 mg, the inelizumab dose may be administered once a month. If the inelizumab dose is 500 mg or 750 mg, the inelizumab dose may be administered once every other week, or alternatively, twice a month. In some aspects, the subject receives 0-2, 0-3, 0-5, 0-10, 1-3, 1-5, or 1-10 administrations.

[0084] In some aspects, inelizumab can be administered at a dosage of about 300 mg. In some aspects, inelizumab can be administered at a dosage of about 250 mg to about 350 mg, about 275 mg to about 325 mg, about 290 mg to about 310 mg, about 205 mg to about 305 mg, or it can be a dosage of 300 mg. In some aspects, the subject can receive one or more initial doses of inelizumab. In some aspects, the subject can receive one, two, three or more initial doses. In some aspects, the initial dose can be about 300 mg. In some aspects, inelizumab can be administered at an initial dose of about 250 mg to about 350 mg, about 275 mg to about 325 mg, about 290 mg to about 310 mg, about 205 mg to about 305 mg, or an initial dose of 300 mg. In some aspects, inelizumab can be administered intravenously in a first initial dose of about 300 mg, a second initial dose of about 300 mg two weeks after the first initial dose, and subsequent doses of about 300 mg every 6 months after the first initial dose.

[0085] The dosage of inelizumab that can be used to treat the method for the subject in need can be a dosage administered intravenously at intervals of about once every 6 months, or once every 7 months, or once every 8 months, or once every 9 months, or once every 10 months, or once every 11 months, once a year, and / or indefinitely. In some aspects, the inelizumab administered in the method disclosed herein can be administered at intervals of about every 6 months. About every 6 months can be administered every 6 months, every 180 days, between every 170 days and 190 days, between every 175 days and 185 days, between every 175 days and 190 days, or between every 170 days and 185 days. About every 6 months can be administered every 26 weeks, every 25 weeks, every 27 weeks, between every 25 weeks and 27 weeks, between every 25 weeks and 26 weeks, or between every 26 weeks and 27 weeks. In some aspects, administration continues indefinitely. Prior to administering inelizumab every approximately 6 months in the methods disclosed herein, an initial inelizumab dose may be administered to a subject in need thereof. The initial inelizumab dose may be administered approximately 2 weeks prior to inelizumab administration approximately every 6 months. Administering the initial inelizumab dose approximately 2 weeks prior to inelizumab administration approximately every 6 months may be administering the initial inelizumab dose 12 days, 13 days, 14 days, 15 days, or 16 days prior to inelizumab administration approximately every 6 months.

[0086] The dose and dosing schedule of inelizumab may be such that any therapeutic effect achieved by administering inelizumab to treat any autoimmune / inflammatory disease or disorder may be "durable." A "durable" effect of inelizumab in treating an autoimmune / inflammatory disease or disorder is a "durable" effect in which the therapeutic effect achieved by inelizumab is maintained (despite no further administration of inelizumab) for at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, or at least 24 weeks after the last dose of inelizumab has been administered. A course of treatment with inelizumab can be a dose of between 500 mg and 3000 mg (e.g., 500 mg, 750 mg, 1000 mg, 1250 mg, 1500 mg, 1750 mg, 2000 mg, 2250 mg, 2500 mg, 2750 mg, or 3000 mg) of inelizumab administered at a dosing interval of once every 7 to every 31 days (e.g., every 7 days, every 10 days, every 14 days, every 15 days, every 16 days, every 14-10 days, every 14-16 days, or every 30 days) over a period of approximately between 8 and 24 weeks (e.g., 8 weeks, or 10 weeks, or 12 weeks, or 14 weeks, or 16 weeks, or 18 weeks, or 20 weeks, or 22 weeks, or 24 weeks, or 2 months, or 4 months, or 6 months).

[0087] In some aspects, administration is intravenous. In some aspects, administration is by infusion.

[0088] The pharmaceutical composition may include an immunosuppressant. Immunosuppressive therapy may include administration of one or more immunosuppressants, such as cytokine transcription inhibitors (e.g., cyclosporine A, tacrolimus), nucleotide synthesis inhibitors (e.g., azathioprine, mycophenolate mofetil), growth factor signal transduction inhibitors (e.g., sirolimus, rapamycin) and T cell interleukin 2 receptor inhibitors (e.g., daclizumab, basiliximab). In a particular embodiment, the immunosuppressant used in combination with the compositions and methods of the present invention includes one or more of the following: adriamycin, azathioprine, busulfan, cyclophosphamide, cyclosporine A ("CyA"), cytotoxin, fludarabine, 5-fluorouracil, methotrexate, mycophenolate mofetil (MOFETIL), nonsteroidal anti-inflammatory drugs (NSAID), rapamycin and tacrolimus (FK506). Immunosuppressants may also include complement inhibitors, such as soluble complement receptor 1, anti-C5 antibodies, or small molecule inhibitors of CI, such as described by Buerke et al. (J. Immunol, 167:5375-80 (2001)).

[0089] In some aspects, the pharmaceutical composition comprises an immunosuppressant in an amount that is effective to immunosuppress the treated subject. In some aspects, the subject is pretreated or has previously received an immunosuppressant prior to treatment with inelizumab. Any immunosuppressant may be administered. In some aspects, the subject has previously been treated with one or both of azathioprine (AZA) and / or mycophenolate mofetil (MMF).

[0090] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific aspects described herein. Such equivalents are intended to be encompassed by the following claims.

[0091] Incorporated by Reference

[0092] This patent application incorporates by reference the following patent publications in their entirety for all purposes: WO2008031056 and WO 2020219743.

[0093] All publications, patents, and patent applications mentioned in this specification are incorporated into the specification by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference.

[0094] Numbered Examples

[0095] Example 1. A method of treating or preventing neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need thereof having a B cell level of less than about 4 cells / μL of blood, thereby treating the NMOSD.

[0096] Embodiment 2. The method of embodiment 1, wherein the Fcγ receptor IIIa (FCGR3A) gene of the subject in need comprises the rs396991 polymorphism.

[0097] Embodiment 3. The method of embodiment 2, wherein the subject in need thereof comprises a V allele determined by a V / F or V / V genotype.

[0098] Example 4. A method of treatment, the method comprising analyzing a blood sample from a subject in need thereof to determine the presence of the rs396991 polymorphism of the FCGR3A gene, and if the rs396991 polymorphism is determined to be positive, administering inerizumab to the subject in need thereof.

[0099] Embodiment 5. The method of embodiment 4, wherein the subject in need thereof suffers from an autoimmune disease.

[0100] Embodiment 6. The method of embodiment 5, wherein the autoimmune disease is selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

[0101] Embodiment 7. The method of embodiment 6, wherein the autoimmune disease is NMOSD.

[0102] Embodiment 8. The method of embodiment 6, wherein the autoimmune disease is MG.

[0103] Embodiment 9. The method of embodiment 6, wherein the autoimmune disease is IgG4RD.

[0104] Embodiment 10. The method of embodiment 4, wherein the subject in need thereof suffers from at least two autoimmune diseases.

[0105] Embodiment 11. The method of embodiment 10, wherein the at least two autoimmune diseases are selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

[0106] Embodiment 12. A method for treating neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need comprising the rs396991 polymorphism of the FCGR3A gene, wherein the administration is more effective in reducing the level of B cells or plasma cells in the subject in need comprising the rs396991 polymorphism of the FCGR3A gene, compared to an otherwise comparable subject lacking the rs396991 polymorphism.

[0107] Embodiment 13. The method of embodiment 12, wherein the level of B cells or plasma cells is determined at least 1, 2, 4, 8, 12, 16, 22, 28 and / or 32 weeks after the administration.

[0108] Embodiment 14. The method of any one of embodiments 1-13, wherein the administration is effective to reduce a criterion selected from the group consisting of: annualized attack rate (AAR), incidence of disability assessed by Expanded Disability Status Scale (EDSS) score, incidence of hospitalization, magnetic resonance imaging (MRI) lesions, and combinations thereof.

[0109] Embodiment 15. The method of embodiment 14, wherein the administration is effective to reduce the AAR, wherein the reduction is at least about 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or up to about 100%.

[0110] Embodiment 16. A method as described in Embodiment 14, wherein the administration is effective to reduce the MRI lesions, wherein the reduction is at least about 20%, 30%, 40%, 50%, 60%, 70%, 73%, 75%, 80%, 85%, 90%, 95%, 97% or up to about 100%.

[0111] Embodiment 17. The method of embodiment 14, wherein the administering is effective to reduce EDSS.

[0112] Embodiment 18. The method of embodiment 14, wherein the administration is effective to reduce the incidence of hospitalization.

[0113] Embodiment 19. The method of any one of embodiments 1-18, wherein the administering is repeated.

[0114] Embodiment 20. The method of embodiment 19, wherein the administration is repeated daily, monthly or yearly.

[0115] Embodiment 21. The method of embodiment 20, wherein the administering is repeated monthly.

[0116] Embodiment 22. The method of embodiment 21, wherein the administration is repeated every 6 months.

[0117] Embodiment 23. The method of any one of embodiments 19-22, wherein the administration continues for at least about 1 year, 2 years, 3 years, 4 years, 5 years, or up to about the lifetime of the subject in need thereof.

[0118] Embodiment 24. The method of any one of embodiments 1-23, wherein the subject in need thereof was previously administered inelizumab.

[0119] Embodiment 25. The method of any one of embodiments 1-24, wherein the administration is intravenous.

[0120] Embodiment 26. The method of embodiment 25, wherein the administering is by intravenous infusion.

[0121] Embodiment 27. The method of any one of embodiments 1-26, wherein the inerizumab is administered at a dose of about 300 mg.

[0122] Embodiment 28. A method of treating or preventing an autoimmune disease, the method comprising administering 300 mg of inerizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months.

[0123] Embodiment 29. The method of embodiment 28, wherein the administering is repeated every 6 months for at least about 1 year.

[0124] Embodiment 30. The method of any one of embodiments 28-29, wherein the autoimmune disease is selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

[0125] Embodiment 31. The method of embodiment 30, wherein the autoimmune disease is NMOSD.

[0126] Embodiment 32. The method of embodiment 30, wherein the autoimmune disease is MG.

[0127] Embodiment 33. The method of embodiment 30, wherein the autoimmune disease is IgG4RD.

[0128] Embodiment 34. A method of treating or preventing neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering 300 mg of inelizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months.

[0129] Embodiment 35. A method of treating or preventing an autoimmune disease, the method comprising administering 300 mg of inelizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months, and wherein the autoimmune disease is selected from the group consisting of: neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

[0130] Embodiment 36. The method of any one of embodiments 34-35, wherein the administering continues for at least about 2.5 years.

[0131] Embodiment 37. A method of treating neuromyelitis optica spectrum disorder, the method comprising administering inelizumab to a subject in need thereof, wherein the subject does not have anti-aquaporin 4 immunoglobulin G antibodies (AQP4-IgG-) and has immunoglobulin G antibodies to myelin oligodendrocyte glycoprotein (MOG-IgG+).

[0132] Embodiment 38. The method of embodiment 37, wherein about 300 mg of the inerizumab is administered.

[0133] Embodiment 39. The method of any one of embodiments 37-38, wherein the administration continues for at least 6 months, 1 year, 2 years or 3 years.

[0134] Embodiment 40. The method of embodiment 39, wherein the administering continues for at least 2 years.

[0135] Embodiment 41. The method of any one of embodiments 1-40, wherein the subject in need thereof has increased serum autoreactivity to autoantibodies not related to NMOSD compared to healthy control subjects.

[0136] Examples

[0137] Example 1 - Safety and efficacy of inelizumab as a treatment for newly presented neuromyelitis optica spectrum disorder

[0138] The study was completed to evaluate the safety and efficacy of inerizumab in newly diagnosed subjects compared to subjects with ≥2 prior NMOSD episodes.

[0139] method

[0140] A double-blind, placebo-controlled, randomized, phase 2 / 3 trial evaluating the efficacy and safety of inelizumab in adults with NMOSD was conducted. Figure 1A), and the trial consisted of the following periods: a randomized controlled period (RCP; 3:1 split to inelizumab [intravenous, 300 mg] or placebo) lasting up to 28 weeks or until an adjudicated attack; an optional open-label period (OLP; inelizumab once every 28 weeks) lasting ≥2 years. No immunosuppressants were used except for oral corticosteroids during the first 2 weeks (with 1 week of dose reduction). The study included adults with NMOSD who had received treatment for ≥1 attack in the past year or had received treatment for ≥2 attacks in the past 2 years and had an EDSS score of ≤8.0. The primary endpoint was the time to the first adjudicated attack during the RCP. Secondary endpoints included annualized attack rate (AAR) and disability progression assessed via the Expanded Disability Status Scale (EDSS). Safety assessments included treatment-emergent adverse events (TEAEs) and treatment-emergent AEs of special interest. Subjects who received inelizumab at any point during the study were included in the 'any inelizumab' group for data analysis purposes. For this study, a post hoc analysis was performed on subjects with AQP4+ NMOSD recruited after the first episode.

[0141] Baseline Characteristics and Demographics

[0142] A total of 37 subjects with newly diagnosed AQP4+ NMOSD and 176 subjects with ≥2 pre-study episodes were included in this analysis (Table 2).

[0143] Table 2: Summary of baseline demographics and characteristics

[0144]

[0145] Of these subjects, 37 newly diagnosed subjects and 173 subjects with ≥2 pre-study episodes were included in the respective 'any inelizumab' groups. Subject characteristics were similar between newly diagnosed subjects and subjects with ≥2 pre-study episodes.

[0146] result

[0147] Inerizumab treatment reduces risk of NMOSD flare-ups

[0148] All subjects treated with inelizumab had a reduced risk of seizures. Newly diagnosed subjects treated with inelizumab had a lower adjudicated seizure incidence, lower AAR, and higher probability of remaining seizure-free at the end of RCP compared with subjects in the placebo group, regardless of the number of seizures before the study. Figure 1B and Figure 1C, Table 3). Similar results were observed in subjects with ≥2 pre-study episodes. In OLP, AAR continued to decrease over time with inelizumab use; AAR was lower for newly diagnosed subjects with 'any inelizumab' and subjects with ≥2 pre-study episodes.

[0149] Table 3: Summary of results

[0150]

[0151] Inelizumab treatment stabilizes EDSS exacerbation

[0152] During RCP, among newly diagnosed subjects and subjects with ≥2 pre-study episodes, fewer subjects had EDSS exacerbations under inelizumab treatment compared with placebo (Table 4).

[0153] Table 4: Summary of results

[0154]

[0155] EDSS was stable in both groups throughout the OLP.

[0156] Safety and tolerability of inelizumab treatment

[0157] During the RCP period, the rates of treatment-emergent AEs and AESIs were no higher in newly diagnosed subjects than in subjects with ≥2 pre-study episodes when divided by treatment group (Table 5).

[0158] Table 5: Summary of treatment-emergent AEs and AESIs occurring during RCP and after any exposure to inelizumab during the entire study period in newly diagnosed subjects and subjects with ≥2 prior relapses by treatment group

[0159]

[0160]

[0161] In conclusion, inelizumab was effective and generally well tolerated in newly diagnosed AQP4+ adults, similar to that observed in adults previously treated with other immunotherapies. In both subgroups, subjects who received inelizumab had fewer flares, a higher probability of remaining flare-free, and fewer EDSS exacerbations compared with those who received placebo.

[0162] Example 2 - The extent of B cell depletion is associated with reduced disease activity in neuromyelitis optica spectrum disorders

[0163] We characterized the relationship between B-cell depletion and outcomes in subjects receiving long-term inelizumab in a double-blind, placebo-controlled, phase 2 / 3 trial.

[0164] method

[0165] The study included a randomized, placebo-controlled period (RCP) of up to 28 weeks, followed by an optional open-label period (OLP) of at least 2 years. B-cell counts were determined using high-resolution flow cytometry (limit of quantification, 0.2 cells / μL). Disease activity was measured using the annualized attack rate (AAR) and the number of new / enlarging T2 lesions.

[0166] result

[0167] Compared with placebo, inelizumab induced rapid B-cell depletion that was sustained through multiple dosing (normal range, 74.4 441.1 cells / μL; mean ± standard deviation [SD], RCP week 4 [W4]: inelizumab, 6.1 ± 9.6 cells / μL; placebo, 147.7 ± 89.4 cells / μL; OLP W156: inelizumab, 10.4 ± 37 cells / μL). B-cell levels at the end of the first inelizumab dosing period (W28) predicted stable, profound depletion with long-term exposure. At W28, clinical and imaging measures of disease activity were lower in subjects with BL (n=139 / 200) compared with subjects with >4 cells / μL (n=61 / 200); rate ratio (95% confidence interval [CI]) for AAR was 0.4 (0.16 0.98), p=0.049, and for new / enlarging T2 lesions was 0.36 (0.23 0.56), p<0.0001. AAR (mean [SE]) was lower in both groups than in the RCP placebo group; L: AAR, 0.03 (0.02 0.04); new / enlarging T2 lesions, 0.49 (0.43 0.56); >4 cells / μL: AAR, 0.09 (0.06 0.12); new / enlarging T2 lesions, 1.36 (1.12 1.61); Placebo: AAR, 1.01 (0.79 1.23); new / enlarging T2 lesions, 2.72 (1.99 3.46). After 2.5 years, long-term inelizumab treatment provided sustained B-cell depletion and reduced indicators of NMOSD disease activity, with significant reductions in AAR (97%) and new / enlarging T2 lesions (73%) relative to the RCP placebo group (p < 0.05 for both).

[0168] Inelizumab provided rapid and durable B-cell depletion in subjects with NMOSD. These findings suggest that deep, durable B-cell depletion is beneficial in NMOSD and that monitoring B-cell counts can inform inelizumab treatment.

[0169] Example 3 - Subjects with neuromyelitis optica spectrum disorder exhibit hallmarks of systemic autoimmunity: widespread serum autoreactivity to nuclear antigens and elevated interferon-inducible gene expression

[0170] A study was completed to dissect the serum autoreactivity and type 1 interferon-inducible gene signature (IFNGS) in the blood and characterize its relationship with disease activity in individuals with NMOSD in this study.

[0171] method

[0172] The study was a multicenter, double-blind, randomized, placebo-controlled, Phase 2 / 3 trial testing the efficacy and safety of inelizumab, a humanized, affinity-optimized anti-CD19 monoclonal antibody, for the treatment of NMOSD. The study included a randomized controlled period (RCP) of up to 28 weeks, followed by an optional open-label period of at least 2 years. Serum samples were obtained from: 220 / 230 subjects with NMOSD randomized in the study, and 25 healthy controls (HC) collected from independent commercial sources. Baseline samples were hybridized to autoantigen microarrays to profile serum immunoglobulin (Ig)G, IgM, IgA, and IgE autoreactivity against the following 122 autoantigens (excluding AQP4) associated with human autoimmune diseases: CENP-A, CENP-B, core histones, DNA polymerase β (POLB), dsDNA, EBNA1, genomic DNA, histone H1, histone H2A, histone H2B, histone H3, histone H4, Jo-1, KU (P70 / P80), La / SSB, Mi-2, PCNA, PL-7, PL-12, PM / Scl-75, PM / Scl 100, ribophosphoprotein P0, ribophosphoprotein P1, ribophosphoprotein P2, Ro / SSA (52Kda), Ro / SSA (60Kda), S100, Scl-70 / topoisomerase I, Sm, Sm / RNP, SmD, SmD1, SmD2, SRP54, ssDNA, ssRNA, U1-snRNP 68 / 70, U1-snRNP A, U1-snRNP B / B, U1-snRNP C, total histone (Histone-total), total histone citrullinated human genomic DNA, KU (P70 / P80), Nup 62, Ro / SSA (52+60) and U1-snRNP. Autoreactivity was confirmed by a signal that was more than five standard deviations above the HC mean. The autoantibody score was calculated by summing the number of points for signals that were more than five standard deviations above the HC mean. Baseline IFNGS from blood RNA samples of 219 subjects with NMOSD and 10 HCs were analyzed simultaneously. IFNGS positivity was confirmed by a change in signal four times higher than the HC mean. Negative binomial regression was used to estimate the possible association between baseline autoreactivity and IFNGS positivity and pre-study NMOSD disease activity. In subjects with a history of more than 4 years, disease activity was measured by the annualized attack rate (AAR) between diagnosis and study baseline or the average annual disability deterioration based on the Expanded Disability Status Scale (EDSS).

[0173] result

[0174] Baseline autoreactivity in subjects with NMOSD

[0175] At baseline, 197 of 220 subjects with NMOSD (90%) had serum autoreactivity to at least one autoantigen, primarily ribonucleoproteins (RNPs), consistent with previous findings ( Figure 2A ). In contrast, HC did not show autoreactivity to any of the antigens studied. Serum autoreactivity was observed mainly in the IgG subclass (83% [n=183]).

[0176] Baseline IFNGS in Subjects with NMOSD

[0177] A total of 27% of subjects with NMOSD (59 / 219) were IFNGS positive at baseline (>4-fold change relative to HC mean) ( Figure 2B A higher proportion of IFNGS-positive subjects were reactive to one or more RNPs than were IFNGS-negative subjects (68% vs. 46%) ( Figure 2C A statistically significant difference in ANA scores between IFNGS-positive subjects and IFNGS-negative subjects was observed ( Figure 2D ). IgG RNP immunoreactive subjects had increased IFNGS at baseline.

[0178] Prior diagnosis of NMOSD and inflammatory disease

[0179] As the researchers report, the majority of subjects with NMOSD who had high autoantigen immune reactivity and positive IFNGS had not been previously diagnosed with other comorbid inflammatory diseases ( Figure 2E Overall, 23 subjects (10%) were diagnosed with a systemic inflammatory disease other than NMOSD. Eight subjects (4%) were previously diagnosed with two or more systemic inflammatory diseases other than NMOSD (Table 6).

[0180] Table 6: Subjects diagnosed with at least two coexisting inflammatory diseases other than NMOSD

[0181] Subject 1 pSS,MG Subject 5 pSS,MG Subject 2 SLE, RA Subject 6 Fibromyalgia, thyroid Subject 3 pSS, thyroid Subject 7 SLE, MG Subject 4 RA, thyroid Subject 8 pSS, thyroid

[0182] Baseline Autoantibodies and IFNGS Positivity and Baseline NMOSD Disease Activity IgG autoantibodies and IFNGS positivity were not associated with increased NMOSD disease activity before the study (Tables 7 and 8).

[0183] Table 7: Autoantibody positivity, IFNGS relative to pre-study AAR in subjects with a history of more than 4 years

[0184]

[0185] Table 8: Autoantibody positivity, IFNGS relative to pre-study EDSS progression in subjects with a history of more than 4 years

[0186]

[0187] Similar results were observed for IgM, IgA, and IgE autoantibodies. In summary, subjects with NMOSD showed higher serum autoreactivity to autoantibodies known to be associated with other systemic autoimmune diseases compared to HC. In addition, subjects with NMOSD and elevated serum immunoreactivity to at least one RNP tended to show increased blood IFNGS at baseline. Most subjects with NMOSD who had high serum immunoreactivity to autoantigens and high blood IFNGS did not have coexisting autoimmune diseases. Neither baseline serum autoreactivity nor IFGNS positivity appeared to be associated with increased NMOSD disease activity.

[0188] Example 4 - The extent of B cell depletion is associated with reduced disease activity in neuromyelitis optica spectrum disorders

[0189] The study was completed to characterize the relationship between B cell depletion and outcomes in subjects in the study who received long-term inelizumab.

[0190] This study characterized the relationship between B cell, plasma cell, and immunoglobulin depletion after inelizumab treatment and longitudinal reductions in NMOSD disease activity after inelizumab treatment.

[0191] method

[0192] Peripheral blood (PB) B-cell and plasma cell profiles and immunoglobulin levels were assessed regularly throughout the study and analyzed for correlation with clinical parameters and magnetic resonance imaging (MRI) lesion activity.

[0193] The study consisted of a randomized controlled period (RCP) of up to 28 weeks, in which subjects were assigned (3:1) to intravenous inelizumab 300 mg or placebo, and an optional open-label period (OLP), during which all subjects received intravenous inelizumab 300 mg every 6 months for at least 2 years ( Figure 1 ).

[0194] Evaluation of B cell pharmacodynamics

[0195] During the RCP period, during the study visits, blood samples were collected at baseline and at weeks 1, 2, 4, 8, 12, 16, 22, and 28 to assess B cell counts; blood samples were collected at baseline and at weeks 2, 4, 8, 12, 16, and 28 to assess plasma cell-specific gene expression; and blood samples were collected at weeks 12 and 28 to assess Ig levels. During the OLP period, blood samples were collected every 13 weeks to assess B cell counts, plasma cell-specific gene expression, and Ig levels. Blood samples were also collected during any assessment visits for new or worsening NMOSD symptoms during the RCP and OLP periods to assess B cell counts and plasma cell gene expression. Samples for attack assessment were collected before starting treatment for an NMOSD attack.

[0196] Within 72 hours of sample collection, in the central laboratory, the B cell level in peripheral blood was analyzed by flow cytometry using fluorescence activated cell sorting (FACS). Whole blood samples were collected in Streck Cytochex cell preservation tubes and transported to the central laboratory on the same day of collection. Minimum 100,000 lymphoid events (lymph event) were obtained. CD20 was used as FACS marker (the combined inelizumab interfered with the FACS based on CD19) to count B lineage cells. The B cell subsets measured included CD20+B cells defined as CD45hi[CD3-, CD56-, CD14-], CD33-, CD20. In addition, FACS was used to analyze plasmablasts / plasma cell counts (CD45hi[CD3-, CD14-, CD56-], CD27+, HLA-DRhi / low, CD38+) to a group of subjects. The lower limit of quantification (LLOQ) of all cell colonies was set to 0.2 cells / μL. Samples with cell counts less than the LLOQ for any given cell population were estimated to be 0.05 cells / μL.

[0197] Plasma cell specific gene expression is assessed by quantitative reverse transcription polymerase chain reaction of blood RNA sample.Plasma cell gene signature is based on the expression analysis of four genes (IGHA1, IGJ, IGKV4-1 and TNFRSF17) mainly expressed by the plasma cells in the blood 21.Feature calculation is the average expression of four plasma cell specific genes at each time point minus the average expression of five control genes (B2M, GAPDH, TFRC, GUSB and UBC).The multiple change of the plasma cell gene expression signature at each time point is calculated relative to a group of ten healthy donor samples and interpreted as the plasma cell abundance relative to the expected prevalence rate under the non-activated immune state.

[0198] TaqMan single nucleotide polymorphism profiling was used to investigate the possible association between the rs396991 polymorphism of the FCGR3A gene and impaired B cell depletion in a group of subjects who consented to genomic analysis. The rs396991 polymorphism is widespread and encodes a valine / phenylalanine substitution at position 158 of FCGR3A, which is associated with reduced binding affinity for Ig Fc and thus impairs the ADCC mechanism optimized by the Fc region of inebilizumab.

[0199] Results

[0200] Compared with placebo, inebilizumab provided rapid B cell and plasma cell depletion (reference range of standard concentration, 74 - 441 cells / μL), which persisted with multiple administrations ( Figure 3A ). B cells decreased in all subjects at 1 week after the first treatment. At week 4 of the RCP, the median (interquartile range [IQR]) B cell count in the inebilizumab group was 2.5 (1.0 - 7.6) cells / μL and in the placebo group was 112.3 (96.3 - 176.9) cells / μL. At week 156 of the OLP, the median (IQR) B cell count with inebilizumab was 0.33 (<LLoQ - 1.0) cells / μL.

[0201] Effect of inebilizumab on long - term B cell depletion

[0202] B cell levels at the end of the first inebilizumab dosing period (week 28) predicted stable, profound depletion with long - term exposure ( Figure 3B ). Among 200 subjects, 70% (n = 139) had B cell depletion to ≤4 cells / μL and had persistent B cell depletion after multiple doses of inebilizumab. Subjects with B cell counts ≤4 cells / μL had persistent, more profound B cell depletion compared with subjects with B cell counts >4 cells / μL ( Figure 3B ). Persistent B cell depletion was also observed in the >4 cells / μL subgroup on continued inebilizumab treatment.

[0203] During the RCP, inebilizumab treatment significantly reduced the circulating levels of B cells and plasma cells compared with placebo ( Figure 3A 、 Figure 6A-6F ). Total Ig levels also decreased, with the greatest decreases observed in the IgE, IgA, and IgM classes. Overall NMOSD disease activity, measured by annual attack rate [AAR], EDSS worsening, NMOSD - related hospitalizations, and new or enlarging T2 MRI lesions, was significantly reduced during RCP18 with inebilizumab treatment and continued to decline with additional doses of inebilizumab ( Figure 7A-7DAfter 2.5 years of inelizumab treatment, AAR, annualized rates of new / enlarging T2 lesions, EDSS worsening, and NMOSD-related hospitalizations gradually decreased.

[0204] Association between B cell dynamics and long-term NMOSD outcomes

[0205] Because long-term inelizumab treatment leads to a gradual decrease in NMOSD disease activity, it was hypothesized that the depth of B-cell depletion would be associated with treatment efficacy. CD20+ peripheral blood B-cell counts were examined during the attack-free, pre-attack, attack, and post-attack periods in subjects with or without attacks at RCP and OLP examinations. Figure 8A-8C No significant increase in peripheral blood CD20+ B cell levels at flare was observed in samples from all subjects or from a subset of subjects who received 3 or more doses of inelizumab ( Figure 8A-8C and Figure 12A-12B ). Regardless of whether the attack occurred during or after the first dosing period, no significant increase in B cell counts was observed during the attack assessment period.

[0206] At 6 months, clinical and imaging measures of NMOSD disease activity were generally lower in subjects with B cell counts ≤4 cells / μL (n / N = 139 / 200) compared with subjects with B cell counts >4 cells / μL (n / N = 61 / 200) ( Figure 3C-Figure 3F Subsequent doses of inelizumab further reduced NMOSD activity measures over time in both subgroups, with a more rapid decline in NMOSD activity measures in the subgroup of subjects with ≤4 cells / μL. All subjects showed similar levels of reduction in NMOSD activity measures after 2.5 years of inelizumab exposure, regardless of B cell counts after the first dosing interval.

[0207] Therefore, we investigated whether the extent of B cell depletion within the first dosing interval could predict the subsequent reduction in the depth of B cell depletion and NMOSD disease activity. Negative binomial regression showed a linear association between B cell counts at the end of the first 6-month dosing interval and disease activity occurring after the first dosing period with inelizumab (Table 9). In our regression analysis, multiple additional time points beyond 6 months were also associated with future disease activity. In fact, B cell counts from samples drawn as early as 3 months after the start of inelizumab treatment were associated with future NMOSD disease activity after the first dosing interval. However, B cell counts from these early time points were not associated with NMOSD disease activity during the first dosing period of treatment.

[0208] Table 9: 6-month B-cell counts relative to long-term disease activity

[0209]

[0210] Negative binomial regression analysis of B-cell counts at 6 months post-treatment (W28 RCP for inelizumab-treated subjects who completed the RCP and W26 OLP for subjects randomized to placebo and inelizumab-treated subjects who withdrew from the RCP early) relative to the annualized rate of flares, new / enlarging T2 lesions, EDSS worsening (subject / year), and hospitalizations occurring after the first dosing period of inelizumab treatment.

[0211] A cutoff of 4 cells / μL separated subjects at risk for decreased NMOSD activity during subsequent inelizumab dosing. Sensitivity analysis showed that a cutoff of 4 cells / μL was the highest threshold that maintained favorable rate ratios across multiple NMOSD activity markers ( Figures 13A-13D Overall, 139 of 200 subjects (70%) had B-cell counts ≤ 4 cells / μL at the end of the first dosing interval and maintained durable B-cell depletion with continued treatment ( Figure 9A-9B When compared with subjects with B cell counts >4 cells / μL, subjects with B cell counts ≤4 cells / μL had consistently lower B cell numbers ( Fig. 9B ), lower AAR (0·034 vs. 0·086; p=0·0494), fewer new / enlarging T2 lesions (0·49 vs. 1·36; p<0·0001), smaller EDSS worsening (0·076 vs. 0·14; p=0·09) and a tendency to fewer hospitalizations (0·08 vs. 0·18; p=0·11; Table 10). Similar disease activity results were observed when the AQP4 seropositive subject population was analyzed separately (Table 10, Figures 14A-14D ).

[0212] Table 10: Long-term outcomes for subjects with B cell counts ≤4 vs. >4 cells / μL at 6 months

[0213]

[0214] With continued inelizumab administration, persistent B cell depletion was still observed in the >4 cells / μL subgroup ( Fig. 9B ), median CD20+ B cell counts were similar between subjects in the >4 cells / μL and ≤4 cells / μL groups through week 117.

[0215] Notably, no increase in overall B cell counts was observed during episodes occurring during this time frame ( Figure 8A-8C and Figure 12A-12B ).

[0216] Effect of long-term inelizumab on disease activity in NMOSD

[0217] Because NMOSD disease activity decreased longitudinally with long-term inelizumab treatment, short- and long-term relationships between clinical and imaging measures and subgroups with 6-month B-cell counts >4 cells / μL or ≤4 cells / μL were assessed. During RCP, NMOSD disease activity (measured by AAR, new / enlarging T2-hyperintense MRI lesions, worsening EDSS scores, and hospitalization rates) decreased in both subgroups after the first dosing period of inelizumab treatment compared with placebo. Subsequent doses of inelizumab further reduced disease activity in both subgroups over time. NMOSD activity decreased more rapidly in subjects with ≤4 cells / μL, especially new T2-hyperintense lesions, compared with subjects in the >4 cells / μL subgroup. However, after 2.5 years of inelizumab exposure, all subjects showed similar levels of NMOSD activity ( Figure 3C-Figure 3E and Fig.10 ).

[0218] After 2.5 years, treatment with inelizumab was associated with reduced NMOSD disease activity compared with placebo during RCP ( Figure 3G ). AAR decreased by 97%. Annualized rate of new or enlarging T2 lesions decreased by 73%.

[0219] In conclusion, treatment with inelizumab provided rapid and durable B-cell depletion in subjects with NMOSD compared with placebo. Greater B-cell depletion was associated with improved outcomes with long-term treatment with inelizumab. These findings suggest, at a minimum, that deep and durable B-cell depletion is beneficial in preventing flares in subjects with NMOSD and / or that regular monitoring of B-cell counts may provide a valuable approach to ensure the most effective use of inelizumab in the management of NMOSD.

[0220] Assessment of Subject Subgroups

[0221] The following three independent but not mutually exclusive mechanisms may affect B-cell counts at the end of the first inelizumab dosing interval:

[0222] Loss of pharmacokinetics of inelizumab may lead to B cell recruitment

[0223] First, the pharmacokinetic (PK) loss of inelizumab after administration of two loading doses may have resulted in B-cell replenishment in some subjects at the end of the first dosing interval. At the end of the first dosing interval, the PK profiles of inelizumab were compared between subgroups with B-cell counts >4 cells / μL and ≤4 cells / μL. Relative to subjects with stable B-cell depletion, subjects with less B-cell depletion had lower inelizumab concentrations: inelizumab was undetectable in the serum of the majority of subjects with B-cell counts >4 cells / μL at week 22 (LLOQ: 100·5 pg / mL, Fig.15 ). Additional analysis of baseline covariates showed that age, sex, race, weight, and BMI were not significantly associated with higher B-cell counts (defined as >4 cells / ul at W28). Prior use of rituximab was also not associated with higher B-cell counts. These subjects showed significantly elevated CD19+ B-cell counts, plasma cell signatures, and Ig concentrations on day 1 of RCP (false discovery rate <0·10) ( Figure 16A-16C ).

[0224] Shallow depletion may be associated with the rs396991 polymorphism in FCGR3A

[0225] Second, the increased B cell counts at the end of the first dosing interval may be caused by shallow cell depletion, which is partially attributed to the rs396991 polymorphism in FCGR3A. In a group of subjects who agreed to genomic analysis, rs396991 F / F homozygosity was analyzed. F / F homozygosity was noted in 100·0% (10 / 10) of subjects who never achieved CD20+≤4 cells / μL during the first dosing interval, 43% (12 / 28) of subjects whose B cell levels were depleted to ≤4 cells / μL but replenished to >4 cells / μL by the end of the first 6-month treatment period, and 45·2% (38 / 84) of subjects with persistent B cell depletion ≤4 cells / μL ( Figure 17A-17B ).

[0226] Anti-drug antibodies may alter B cell exhaustion

[0227] Third, antidrug antibodies (ADA) that bind or neutralize inelizumab may affect changes in B-cell counts. Overall, 33 of 225 subjects (14·7%) treated with inelizumab intermittently tested positive for ADA on study (lowest titer 1:50 dilution; median maximum titer = 100), and these titers generally decreased with long-term inelizumab treatment ( Fig.18AIn general, B cell counts increased slightly with ADA positivity, but no significant overlap was found between subjects who were ADA positive and those with B cell levels > 4 cells / μL before completion of the first dosing interval ( Fig.18B ). Median B-cell counts in 90-day samples drawn during the ADA-positive period were higher than those in samples from subjects who remained persistently ADA-negative (1·5 cells / μL versus 0·4 cells / μL; Fig. 18C ).

[0228] Example 5 - Sustained B cell depletion by inelizumab is associated with reduced disease activity in aquaporin-4 seropositive neuromyelitis optica spectrum disorder

[0229] As previously described in Example 3 and Figure 1, studies were completed to characterize the relationship between B cell depletion and outcomes in subjects receiving long-term inelizumab in the N-MOmentum trial.

[0230] Effect of inerizumab on B cell depletion

[0231] Compared with placebo, inelizumab provides rapid B cell depletion. All subjects had B cell reductions at 1 week after the first treatment; the reduction was maintained over time at multiple doses. After the first dosing interval (6 months), 126 of 186 subjects (68%) had B cell counts of ≤4 cells / μL. The remaining subjects (n / N=60 / 186 [32%]) had >4 cells / μL, but were well below the lower limit of normal. Long-term (>2.5 years) inelizumab treatment provided sustained B cell depletion in all subjects, regardless of the depth of short-term B cell depletion.

[0232] Short-term effects of B cell depletion on markers of disease activity

[0233] Compared with placebo, immediate clinical benefit was observed in both groups; however, during the first dosing interval, subjects with ≤4 cells / μL had greater improvements in disease activity measures than those with >4 cells / μL. After the first dosing interval, 111 of 124 subjects with ≤4 cells / μL (90%) were flare-free and 50 of 59 subjects with >4 cells / μL (85%) were flare-free. Subsequent doses of inerizumab further reduced NMOSD activity measures over time in both subgroups, with a more rapid decline in NMOSD activity measures in subjects with ≤4 cells / μL.

[0234] Table 11: Overview of disease activity indicators

[0235]

[0236]

[0237] Effect of long-term (>2.5 years) inelizumab-associated B-cell depletion on disease activity measures.

[0238] When compared with placebo during RCP, long-term treatment with inelizumab was associated with reduced AAR ( Figure 4A ), reduced EDSS exacerbation in both subgroups ( Figure 4B ), and the depth of B-cell depletion after the first dosing interval was not associated with infection risk ( Figure 4C ).

[0239] In conclusion, inelizumab provided rapid, durable B-cell depletion in subjects with NMOSD. After the first dosing interval, B-cell depletion and subsequent reductions in disease activity varied between subjects. Monitoring B-cell counts could be used to optimize inelizumab treatment. Long-term inelizumab treatment provided deep B-cell depletion and reductions in disease activity in all subjects, regardless of the depth of B-cell depletion during their first dosing period. The depth of B-cell depletion was not associated with infection risk with long-term inelizumab treatment or after the first dosing interval.

[0240] Example 6 - Subjects with neuromyelitis optica spectrum disorder exhibit hallmarks of systemic autoimmunity: widespread serum autoreactivity to nuclear antigens and elevated interferon-inducible gene expression

[0241] Studies were completed to profile serum autoreactivity and type 1 interferon-inducible gene signature (IFNGS) in blood to characterize their relationship with disease activity in NMOSD.

[0242] method

[0243] Serum samples were obtained from 220 subjects of 230 NMOSD subjects randomly grouped in NMOmentum, and 25 healthy controls (HC). The serum immunoglobulin (Ig) G, IgM, IgA and IgE autoreactivity of baseline samples were analyzed by microarray hybridization of baseline samples with 122 autoantigens (excluding AQP4) related to human autoimmune diseases (e.g., systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome). Autoreactivity was confirmed by a signal higher than the HC mean value>5 standard deviations. Baseline IFNGS of blood RNA samples from 219 subjects and 10 HCs was analyzed. Samples with a signal higher than the HC mean value>4 times the change were IFNGS positive. The possible association of baseline autoreactivity and IFNGS positivity with NMOSD disease activity before the study (annual onset rate between diagnosis and baseline or average annual disability deterioration based on the expanded disability status scale, in subjects with a history of>4 years) was studied by negative binomial regression.

[0244] result

[0245] At baseline, HCs did not show autoreactivity to any of the antigens studied, whereas 90% of subjects with NMOSD (n=197) were autoreactive to self-antigens, mostly of the IgG subclass (83% [n=183]), but also of the IgA (41% [n=90]), IgM (25% [n=56]), and IgE (6% [n=14]) subclasses. Overall, 27% of subjects (n=59) were IFNGS-positive at baseline and 4% (n=8) had a previous diagnosis of systemic autoimmune disease. Serum autoreactivity to nuclear antigens (NA) was greater than to other antigens. A higher proportion of IFNGS-positive subjects were reactive to NA than to IFNGS-negative subjects (68% [40 / 59] vs. 46% [71 / 153]; area under the ROC curve, 0.63; p=0.002). Neither serum autoreactivity nor IFNGS positivity at baseline was associated with an increased risk of pre-study NMOSD disease activity.

[0246] Compared with healthy controls, subjects with NMOSD showed higher serum autoreactivity to autoantibodies known to be associated with other systemic autoimmune diseases. However, this autoreactivity did not appear to be associated with increased NMOSD disease activity.

[0247] Example 7 - Effect of low-affinity immunoglobulin gamma Fc region receptor III-A gene polymorphism in neuromyelitis optica spectrum disorder and association with treatment outcome

[0248] A study was conducted to characterize the relationship between the rs396991 polymorphism, NMOSD disease activity, and treatment response.

[0249] method

[0250] The study had a randomized controlled period (RCP; inelizumab 300 mg or placebo on days 1 and 15) of up to 28 weeks, followed by an open-label period (OLP). 142 subjects (104 randomized to inelizumab and 38 randomized to placebo) consented to polymorphism genotyping via TaqMan qPCR assay.

[0251] result

[0252] The change in annualized attack rate (AAR) and Expanded Disability Status Scale (EDSS) score from NMOSD onset to recruitment was nominally higher in subjects with the rs396991 V allele (V; V / V or V / F genotype; n=74) compared with subjects with the F / F allele (F / F; n=68): mean (±SEM) AAR: V, 1.0 (0.8 1.3); F / F, 0.7 (0.5 0.9); change in EDSS score: V, 0.6 (0.5 0.7); F / F, 0.4 (0.3 0.5). In the placebo group, the rates of flare, new / enlarging T2 MRI lesions, and NMOSD-related hospitalizations were higher in V allele subjects (n=22) than in F / F allele subjects (n=16), but not statistically significantly: mean (±SEM) AAR: V, 1.3 (0.9 1.7); F / F, 0.8 (0.5 1.2); new T2 lesions: V, 4.5 (2.6 6.3); F / F, 2.1 (1.0 3.1); hospitalizations: V, 0.5 (0.2 0.8); F / F, 0.2 (0.0 0.4). At the end of RCP, V allele carriers randomized to inerizumab (n=52) had greater median (IQR) B cell, plasma cell, and Ig depletion compared with F / F homozygotes (n=52): B cells: V, 0.6 (0.1 3.2) vs. F / F, 1.3 (0.5 4.2) cells / μl; plasma cells: V, 0.04 (0.02 0.2) vs. F / F, 0.05 (0.03 0.2) fold change from control mean; Ig: V, 1050 (867 1284) vs. F / F, 1238 (1076 1455) Ig / μl. V allele carriers also had lower rates of flare and new / enlarging T2 lesions than F / F homozygotes, but the differences were not statistically significant: mean (± SEM): AAR: V, 0.1 (0.10.2); F / F, 0.3 (0.20.4); new T2 lesions: V, 1.4 (0.91.8); F / F, 1.7 (1.22.2). In OLP, there were few differences in clinical parameters or B-cell depletion between subgroups by dose 4.

[0253] In conclusion, V allele carriers may have increased NMOSD disease activity but may have a higher early pharmacodynamic response to inerizumab compared with those with the wild-type F / F genotype.

[0254] Example 8 - Effect of low-affinity IgG Fc region receptor III-A gene polymorphism on treatment outcomes in neuromyelitis optica spectrum disorder

[0255] A study was conducted to characterize the relationship between the rs396991 polymorphism, NMOSD disease activity, and treatment response.

[0256] method

[0257] The study was a double-blind, randomized, placebo-controlled, Phase 2 / 3 trial of the efficacy and safety of inelizumab in 230 adults with NMOSD. The study included a randomized controlled period (RCP) of up to 28 weeks, in which subjects were assigned (3:1) to intravenous (iv) inelizumab 300 mg or placebo; followed by an optional open-label period of at least 2 years (OLP; all subjects received inelizumab 300 mg iv every 26 weeks). The primary endpoint was the risk of NMOSD attack, assessed by the time to the onset of an NMOSD attack. A total of 142 subjects (inelizumab, n=104; placebo, n=38) consented to polymorphism genotyping via TaqMan quantitative polymerase chain reaction assay.

[0258] In a post hoc analysis, data on disease severity and response to inerizumab were compared between subjects with different FCGR3A genotypes. Outcome analyses included flares, disability (assessed by Expanded Disability Status Scale [EDSS] score), NMOSD-related hospitalizations, and magnetic resonance imaging (MRI) lesions.

[0259] result

[0260] Pre-study disease severity in subjects with V allele

[0261] V allele subjects ( Figure 5B The historical annualized attack rate (AAR) from the onset of NMOSD to recruitment was nominally higher in subjects with V allele (V / F or V / V genotype; n = 25) than in subjects with F / F genotype (F / F; n = 25). Median (interquartile range [IQR]): V allele, 0.9 (0.5-1.4); F / F genotype, 0.4 (0.3-0.8). The change in EDSS score was nominally higher in subjects with V allele than in subjects with F / F genotype ( Figure 5B). Median (IQR): V allele, 0.5 (0.4-0.9); F / F genotype, 0.4 (0.3-0.7). For this analysis, all subjects were assumed to have an EDSS score of zero at the time of NMOSD disease onset. No significant differences were observed across the cohort. Disease activity in subjects with the V allele in the study. In RCP, subjects with the V allele who received placebo had higher AARs and rates of new / enlarged T2 MRI lesions and NMOSD-related hospitalizations than those subjects who were homozygous for the F allele (although these differences did not reach statistical significance). The means (± standard error of the mean [SEM]) of these parameters for subjects with the V allele relative to the F / F genotype were as follows: onset, 1.3 (0.9-1.7) vs. 0.8 (0.5-1.2) ( Figure 5C ), new / enlarging T2 lesions, 4.5 (2.6-6.3) vs. 2.1 (1.0-3.1) ( Figure 5C ), NMOSD-related hospitalization, 0.5 (0.2-0.8) vs. 0.2 (0.0-0.4) ( Figure 5C ).

[0262] Effect of inelizumab in subjects with the V allele

[0263] The AAR and annual rate of new / enlarging T2 lesions were lower in V allele subjects treated with inelizumab than in F allele subjects. The means (±SEM) of these parameters for V allele versus F / F genotype subjects were as follows: AAR, 0.1 (0.1-0.2) versus 0.3 (0.2-0.4) ( Figure 5D ), annual rate of new / enlarged T2 lesions, 1.4 (0.9-1.8) vs. 1.7 (1.2-2.2) ( Figure 5D ). However, these differences did not reach statistical significance. B cell, plasma cell, and IgG depletion was greater in V allele subjects treated with inelizumab compared with F / F allele subjects. At the end of RCP, the median values ​​(IQR) for these parameters in V allele subjects (n=52) relative to F / F genotype subjects (n=52) were as follows: B cell depletion, 0.6 (0.1-3.2) vs. 1.3 (0.5-4.2) cells / μL ( Figure 5E ), plasma cell depletion, 0.04 (0.02-0.2) relative to 0.05 (0.03-0.2) fold change from the control mean ( Figure 5E ), IgG level, 1050 (867-1284) vs. 1238 (1076-1455) mg / dL ( Figure 5E ).

[0264] Long-term inelizumab treatment

[0265] In the setting of repeated inelizumab dosing (1.5-2.5 years) in OLP, there were no significant differences in clinical measures of NMOSD activity or B-cell depletion between the subgroups of subjects with the V allele and those with the F / F genotype ( Fig. 5F ).

[0266] In summary, the F176V polymorphism of FCGR3A (V / F or V / V genotype) is not associated with significant differences in NMOSD disease activity compared to the F / F genotype. Subjects with the V allele may have a greater pharmacodynamic response to inelizumab compared to those with the F / F genotype. In subjects treated with inelizumab, V allele subjects tended to have greater B cell and plasma cell depletion and a lower attack rate and fewer new / enlarged T2 lesions. In addition, V allele carriers may have increased NMOSD disease activity compared to F / F allele homozygotes. In this study of subjects treated with inelizumab, no significant differences were observed in the results between those subjects with the F and V allele genotypes.

[0267] Inelizumab reduces the risk of neuromyelitis optica spectrum disorder attacks independent of low-affinity IgG Fc region receptor III-A gene polymorphism

[0268] V allele carriers (V allele genotype [V / V or V / F], n=74) and F / F allele homozygotes (n=68) did not show significant differences in baseline demographics or disease duration. CD20+B cell depletion was similar in V allele vs. F / F subjects (0.6 (0.1-3.2) vs. 1.3 (0.5-4.2) cells / μl at the end of RCP) and continued in both groups throughout the study duration. No differences were found in the risk of relapse (OR 0.94 (0.39, 2.24)) or the expanded disability status scale deterioration (OR: 1.55 (0.54, 4.70)) for V vs. F / F subjects. The annualized attack rate (SEM) for patients treated with inelizumab was V / V 0.00 (0.00), V / F 0.10 (0.04) and F / F 0.06 (0.03). Inerizumab-treated subjects in the N-MOmentum trial showed no differences in clinical outcomes between subjects with the F and V allele genotypes.

[0269] Example 9 - Myelin oligodendrocyte glycoprotein immunoglobulin g in aquaporin-4 seronegative patients with neuromyelitis optica spectrum disorder: results from an inelizumab study

[0270] A study was conducted to report outcomes in subjects with serotype AQP4-IgG- / MOG-IgG+ who received long-term inelizumab in a double-blind, placebo-controlled, Phase 2 / 3 study.

[0271] method

[0272] The study consisted of a randomized controlled period (RCP) of up to 28 weeks, in which subjects were assigned (3:1) to intravenous inelizumab (300 mg) or placebo, followed by an optional open-label period (OLP), in which all subjects received inelizumab for at least 2 years. Fig.19A ). Data for any exposure to inerizumab included time in the RCP and OLP.

[0273] An expert eligibility committee reviewed all screened AQP4-IgG- subjects; subjects who met the 2006 NMOSD diagnostic criteria (Wingerchuk DM et al. Neurology 2006; 66: 1485-9) were randomized.

[0274] End points included: (1) time to onset of NMOSD episodes as determined by an adjudication committee (AC; annualized episode rate [AAR] was also determined); (2) worsening of Expanded Disability Status Scale (EDSS) score; (3) cumulative total number of active lesions or new / enlarging T2 lesions detected by magnetic resonance imaging; (4) number of NMOSD-related hospitalizations; (5) B-cell counts in peripheral blood; (6) serum immunoglobulin levels; (7) adverse events (AEs); and (8) AEs of special interest (AESIs).

[0275] result

[0276] Efficacy in AQP4-IgG- / MOG-IgG+ Subjects

[0277] A total of 17 (7.4%) of the 231 subjects with NMOSD were AQP4-IgG-; of these, 7 (3.0%) were MOG-IgG+ (Inerizumab, n=6; Placebo, n=1). In those subjects randomized to Inerizumab, there were (each occurring in a separate subject) (Table 12, Fig.19B ): one AC-confirmed episode 78 days after the first dose; one disability exacerbation event; and one hospitalization.

[0278] Table 12: Time from first dose of inerizumab to first AC confirmed onset

[0279]

[0280] Data are n (%). EDSS worsening was defined as an increase of ≥2 points from a baseline score of 0, an increase of ≥1 from a baseline score of 1-5, or an increase of ≥0.5 from a baseline score of ≥5.5.

[0281] AC, adjudication committee; EDSS, expanded disability status scale; MRI, magnetic resonance imaging;

[0282] RCP, randomized controlled period.

[0283] The mean (95% confidence interval) AAR for AQP4-IgG- / MOG-IgG+ subjects was 3.12 (1.52-4.72) in the 2 years prior to recruitment; during the study it was 0.05 (0.01-0.35). Inerizumab exposure in RCP and OLP was 20.05 person-years (n=7).

[0284] CD19 B cells and immunoglobulin levels in AQP4-IgG- / MOG-IgG+ subjects

[0285] In the majority of AQP4-IgG- / MOG-IgG+ subjects, B cell depletion under inelizumab was rapid and sustained, see Fig.19C One year after the first dose of inerizumab, at least half of the subjects with serotype AQP4-IgG- / MOG-IgG+ had IgG and IgM levels below the lower limit of normal, see Table 13.

[0286] Table 13: Subjects with baseline serotype AQP4-IgG- / MOG-IgG+ and total serum immunoglobulin levels below the lower limit of normal during the study.

[0287]

[0288] The data are n(%).

[0289] a The lower limit of normal is: IgG, 565mg / dL; IgM, 40mg / dL.

[0290] AQP4, aquaporin-4; Ig, immunoglobulin; MOG, myelin oligodendrocyte glycoprotein.

[0291] Considering any subject exposed to inerizumab, the safety profile was similar to that observed in RCP; one subject had several mild (grade 1) infusion-related reactions (Table 14). No neutrophil- or lymphocyte-related AEs were reported, and no new safety signals were observed.

[0292] Table 14: AEs and AESI during any exposure to inelizumab

[0293]

[0294] The data are n(%).

[0295] AE, adverse event; AESI, adverse event of special interest.

[0296] in conclusion

[0297] These post hoc analyses of the study suggest that inerizumab was generally well tolerated and reduced NMOSD attacks in subjects with NMOSD and serotype AQP4-IgG- / MOG-IgG+.

[0298] Example 10 - Relationship between B cell subsets and aquaporin-4 antibody titers and disease activity in subjects treated with inelizumab

[0299] A study was conducted to assess B cell subsets and AQP4 IgG titers in the peripheral blood of subjects receiving treatment with inerizumab.

[0300] method

[0301] Subjects received 300 mg of inelizumab or placebo (PBO) on days 1 and 15 during the randomized controlled period (RCP) and every 6 months in the optional open-label period (OLP). Absolute counts of CD20+B cells and CD27+ memory B cells were assessed by flow cytometry (RCP+OLP). Plasma cell (PC) gene expression was assessed by qRT-PCR (RCP only). AQP4 IgG titers were determined by cell-based assays (RCP only). All measurements were performed in peripheral blood.

[0302] result

[0303] In the placebo group, a significant increase in CD20+B cells and CD27+ memory B cells was observed at the time of the attack relative to the previous visit (p<0.05). An increase in the PC subset was observed at the previous visit relative to baseline (p<0.01). During the attack, 4 of 20 (20%) observed an increase in CD20+B cells >2-fold from baseline, 3 of 19 (16%) observed an increase in memory B cells >2-fold from baseline, and 11 of 20 (55%) observed an increase in PC >2-fold from baseline. Inelizumab significantly reduced all B cell subsets. No significant increase in any B cell subset was observed at the time of the attack in the inelizumab group relative to the previous visit. There was no significant difference between the treatment groups in the change in AQP4 IgG titer from baseline to the attack (p=0.15). At the end of RCP, 9 of 50 PBO subjects (18%) vs. 59 of 159 inelizumab subjects (37%) (p=0.014) had a ≥2-fold reduction in AQP4-IgG (≥8-fold reduction in 0% vs. 11%, p=0.008).

[0304] Increased levels of B cell subsets (particularly plasma cell subsets) during flares were observed in the placebo group but not the inerizumab group. In a subset of subjects, inerizumab treatment was associated with a reduction in AQP4-IgG.

[0305] Example 11 - Safety and efficacy of inelizumab in AQP4+ NMOSD subjects with a history of immunosuppressive therapy

[0306] The study was completed to evaluate the long-term outcomes of inerizumab treatment in AQP4+ NMOSD subjects with a history of immunosuppressant therapy compared to subjects without a history of immunosuppressant therapy.

[0307] method

[0308] Subjects had previously completed a 28-week randomized Phase 2 / 3 trial of inelizumab versus placebo, and an optional open-label extension (OLE) (>2 years). Immunosuppressant medications for the prevention or treatment of NMOSD relapses were allowed prior to dosing on Day 1. In this post hoc analysis, AQP4+ subjects who received inelizumab (in the OLE) were grouped by no history of immunosuppressive therapy other than treatment of acute NMOSD attacks (naive), or prior azathioprine (AZA) and / or mycophenolate mofetil (MMF) therapy. The outcomes compared between the two groups included annualized relapse and hospitalization rates, as well as safety assessments.

[0309] result

[0310] Of the subjects who received inelizumab during the study, 94 had received prior AZA / MMF and 103 were immunosuppressant-naive. The total number of patient-years of inelizumab treatment was 300.35 in the prior AZA / MMF group and 335.7 in immunosuppressant-naive subjects. The annualized relapse rate (95% confidence interval [CI]) was 0.11 (0.07, 0.17) for subjects with prior AZA / MMF compared with 0.08 (0.05, 0.14) for those without treatment. The annualized NMOSD-related hospitalization rate (annualized rate [95% CI]) was 0.15 (0.08, 0.27) for subjects with prior AZA / MMF and 0.12 (0.06, 0.22) for those without treatment. For subjects with prior AZA / MMF, the percentage of subjects with ≥1 study drug-related treatment-emergent adverse event (TEAE) was 30.9% (29 / 94) and 47.6% (49 / 103) for those without prior AZA / MMF; 4.3% (4) of subjects with prior AZA / MMF and 5.8% (6) of subjects without prior immunosuppressant therapy reported ≥1 study drug-related serious adverse event. Most adverse events in both groups were related to infections; (72.3% (68 / 94) for prior AZA / MMF and 77.7% (80 / 94) for those without prior AZA / MMF).

[0311] In conclusion, post hoc analyses evaluating long-term outcomes of inelizumab in AQP4+ NMOSD subjects treated with prior AZA / MMF therapy demonstrated a similar efficacy and safety profile to subjects without prior immunosuppressant therapy.

Claims

1. A method of treating or preventing neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need thereof having a B cell level of less than about 4 cells / μL of blood, thereby treating the NMOSD.

2. The method of claim 1, wherein the subject in need thereof has an Fcγ receptor IIIa (FCGR3A) gene comprising the rs396991 polymorphism.

3. The method of claim 2, wherein the subject in need thereof comprises a V allele at amino acid position 158 of the FCGR3A gene determined by a V / F or V / V genotype.

4. A method of treatment, the method comprising determining the presence of the rs396991 polymorphism in the FCGR3A gene in a subject in need thereof; and if the rs396991 polymorphism is determined to be positive, administering inelizumab to the subject in need thereof.

5. The method of claim 4, wherein the subject in need thereof suffers from an autoimmune disease.

6. The method of claim 5, wherein the autoimmune disease is selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

7. The method of claim 6, wherein the autoimmune disease is NMOSD.

8. The method of claim 6, wherein the autoimmune disease is MG.

9. The method of claim 6, wherein the autoimmune disease is IgG4RD.

10. The method of claim 4, wherein the subject in need thereof suffers from at least two autoimmune diseases.

11. The method of claim 10, wherein the at least two autoimmune diseases are selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

12. A method for treating neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering inelizumab to a subject in need comprising the rs396991 polymorphism of the FCGR3A gene, wherein the administration is more effective in reducing the level of B cells or plasma cells in the subject in need comprising the rs396991 polymorphism of the FCGR3A gene, compared to an otherwise comparable subject lacking the rs396991 polymorphism.

13. The method of claim 12, wherein the level of B cells or plasma cells is determined at least 1, 2, 4, 8, 12, 16, 22, 28 and / or 32 weeks after the administration.

14. The method of any one of claims 1-13, wherein the administration is effective to reduce a criterion selected from the group consisting of: annualized attack rate (AAR), incidence of disability as assessed by Expanded Disability Status Scale (EDSS) score, incidence of hospitalization, number of lesions detected by magnetic resonance imaging, and combinations thereof.

15. The method of claim 14, wherein the administration is effective to reduce the AAR score, wherein the reduction is at least about 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or up to about 100%.

16. The method of claim 14, wherein the administering is effective to reduce the number of MRI lesions.

17. The method of claim 16, wherein the reduction is at least about 20%, 30%, 40%, 50%, 60%, 70%, 73%, 75%, 80%, 85%, 90%, 95%, 97%, or up to about 100% compared to a comparable untreated subject.

18. The method of claim 14, wherein the administration is effective to reduce the incidence of hospitalization compared to comparable untreated subjects.

19. The method of any one of claims 1-18, wherein the administering is repeated.

20. The method of claim 19, wherein the administration is repeated daily, monthly or yearly.

21. The method of claim 20, wherein the administration is repeated monthly.

22. The method of claim 21, wherein the administration is repeated every 6 months.

23. The method of any one of claims 19-22, wherein the administration continues for at least about 1 year, 2 years, 3 years, 4 years, 5 years, or up to about the lifetime of the subject in need thereof.

24. The method of any one of claims 1-23, wherein the subject in need thereof was previously administered inelizumab.

25. The method of any one of claims 1-24, wherein the administration is intravenous.

26. The method of claim 25, wherein the administering is by intravenous infusion.

27. The method of any one of claims 1-26, wherein the inerizumab is administered at a dose of about 300 mg.

28. A method of treating or preventing an autoimmune disease, the method comprising administering 300 mg of inerizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months.

29. The method of claim 28, wherein the administering is repeated every 6 months for at least about 1 year.

30. The method of any one of claims 28-29, wherein the autoimmune disease is selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

31. The method of claim 30, wherein the autoimmune disease is NMOSD.

32. The method of claim 30, wherein the autoimmune disease is MG.

33. The method of claim 30, wherein the autoimmune disease is IgG4RD.

34. A method of treating or preventing neuromyelitis optica spectrum disorder (NMOSD), the method comprising administering 300 mg of inerizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months.

35. A method of treating or preventing an autoimmune disease, the method comprising administering 300 mg of inelizumab to a subject in need thereof having B cell levels less than about 4 cells / μL of blood, wherein the administration is repeated every 6 months, and wherein the autoimmune disease is selected from the group consisting of neuromyelitis optica spectrum disorder (NMOSD), myasthenia gravis (MG), and IgG4RD.

36. The method of any one of claims 34-35, wherein the administering continues for at least about 2.5 years.

37. A method of treating neuromyelitis optica spectrum disorder, the method comprising administering inelizumab to a subject in need thereof, wherein the subject does not have anti-aquaporin 4 immunoglobulin G antibodies (AQP4-IgG-) and has immunoglobulin G antibodies to myelin oligodendrocyte glycoprotein (MOG-IgG+).

38. The method of claim 37, wherein about 300 mg of the inerizumab is administered.

39. The method of any one of claims 37-38, wherein the administration continues for at least 6 months, 1 year, 2 years or 3 years.

40. The method of claim 39, wherein the administering continues for at least 2 years.

41. The method of any one of claims 1-40, wherein the subject in need thereof has increased serum autoreactivity to autoantibodies not related to NMOSD compared to healthy control subjects.

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