Methods of treatment with IFNβ antibodies

Abstract

The present invention provides methods for treating a patient having one or more conditions associated with IFNβ expression in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen includes multiple individual doses separated by at least one week from each other, as well as formulations suitable for such methods.

Description

[Technical Field]

[0001] The present invention relates to the treatment of signs and symptoms of idiopathic inflammatory myopathy with anti-interferon beta (IFNβ) antibodies. [Background technology]

[0002] Idiopathic inflammatory myopathies (IIM) are a heterogeneous group of rare, chronic autoimmune diseases characterized by muscle inflammation, muscle weakness, and extramuscular manifestations, including lesions of the skin, lungs, heart, gastrointestinal tract, and joints. The most common types of IIM in adults are dermatomyositis (DM), polymyositis (PM), and inclusion body myositis (IBM), and in children, juvenile form of DM (JDM) (Malik et al., 2016). Patients diagnosed with IIM are often classified into subtypes, which allows clinicians to predict prognosis and individualize treatment plans and other therapeutic approaches (Loarce-Martos et al., 2021). The Bohan and Peter criteria, published in 1975, are commonly used to classify patients with DM and PM. Since then, and in light of the discovery and use of autoantibody testing and the identification of distinct subgroups, new classification criteria have been developed (Baig and Paik, 2020). Validated classification criteria were approved by the ACR and EULAR in 2017 and are now widely used and accepted for classifying patients with IIM (Lundberg et al., 2017). These criteria categorize patients as having "definite," "probable," "suspected," and "non-IIM" based on disease scores and corresponding probabilities. Subclassification of IIM distinguishes between adult and juvenile myositis, polymyositis, dermatomyositis, DM without muscle symptoms, or inclusion body myositis.

[0003] DM is a rare, severe, and severely debilitating autoimmune disease with both neuromuscular and cutaneous manifestations. The disease is characterized by skeletal muscle weakness and inflammation of the skin accompanied by a distinct, severe skin rash (Dalakas, 1991). Skin manifestations can occur in up to 94% of patients with DM and are often preceded or accompanied by muscle weakness (Ahmed et al., 2020a). Muscle weakness is also a common feature in more than 80% of DM cases, with patients exhibiting proximal skeletal muscle weakness and elevated serum muscle enzymes, such as creatine kinase and aldolase (Bohan and Peter, 1975a; Bohan and Peter, 1975b; Dalakas and Hohlfeld, 2003; Findlay et al., 2015). Symptoms may develop suddenly or gradually over time, often waxing and waning without apparent cause. DM is also associated with a general increased risk of malignancies, particularly ovarian, lung, pancreatic, gastric, and colorectal cancer, as well as reduced life expectancy (Oldroyd et al., 2021; Vaughan et al., 2022). Other clinical symptoms associated with DM include calcification, cardiac abnormalities (including arrhythmias, congestive heart failure, myocarditis, pericarditis, angina, and fibrosis), dysphagia (difficulty swallowing), polyarthritis, and interstitial lung disease (Na et al., 2009; Khan and Christopher-Stine, 2011).

[0004] Juvenile DM (JDM) is a rare, chronic, often autoimmune disease that begins in childhood. It is characterized by proximal muscle weakness and a pathognomonic skin rash. Although the etiology remains unclear, JDM has been proposed to be caused by vascular damage within the muscle tissue and multiple other organ systems in genetically predisposed individuals in response to environmental triggers. Delayed treatment of JDM can lead to poor outcomes in terms of disease course and calcification (Batthish and Feldman, 2011). There are several differences between juvenile and adult DM in terms of characteristics, outcomes, and prevalence of comorbidities. Children with DM have more vascular damage, calcification, periungual and gingival telangiectasia, and ulceration, but have improved survival and a better long-term prognosis compared with adult DM. Adults with DM are more likely to have myositis-specific antibodies, develop ILD, have disease without muscle symptoms, and have a significant association with malignancy ( Oldroyd et al., 2021 , Vaughan et al., 2022 ) and other comorbidities ( Robinson and Reed, 2011 ).

[0005] Polymyositis (PM) is a rare disease that primarily affects adults. Since the publication of the Bohan and Peter classification criteria, several classification criteria for IIM have been used, but they share the common limitation of not necessarily excluding other types of myopathy and misclassifying IBM patients as PM. Subgrouping patients based on clinical symptoms, histopathological findings, and the presence of MSA may improve classification accuracy. While the use of MSA is increasing, classification criteria based on these criteria have not been validated. According to the ACR / EULAR criteria, patients classified as PM may have autoantibodies associated with antisynthase syndrome and immune-mediated necrotizing myopathy (Lundberg et al., 2017). This group may also include patients without specific autoantibodies but with clinical symptoms consistent with PM. PM primarily manifests in the muscles, with characteristic proximal muscle weakness, elevated muscle enzymes, and myopathic features on electromyography. Dysphagia may occur in some patients. The most commonly associated extramuscular manifestations are interstitial lung disease and cardiac involvement. Cutaneous manifestations of DM are absent in patients with PM (Malik et al., 2016; Baig and Paik, 2020).

[0006] In terms of epidemiology, DM affects both adults and children (Findlay et al., 2015). DM most commonly occurs after the age of 40 (Bogdanov et al., 2018), and women are twice as likely to be affected as men (NORD, 2015). Juvenile DM is the most common inflammatory myositis in children, most commonly observed between the ages of 5 and 12 (Dourmishev AL, 2009). PM primarily affects adults, with a higher adult-to-child ratio than observed for DM (Shah et al., 2013). PM is also more prevalent in women than men, with estimates ranging from 50% (Svensson et al., 2017) to 2-fold higher in women (See et al., 2013). Juvenile PM is much less prevalent (less than one-tenth of the prevalence) than JDM. The mean age at diagnosis of JPM is 12.1 years, compared with the median age of JDM diagnosis of 7.4 years ( Shah et al., 2013 ).

[0007] From the patient's perspective, a common clinical symptom across all types of IIM is symmetric proximal muscle weakness. Many patients with IIM have extramuscular symptoms as part of their initial clinical manifestations. These include skin rashes, particularly in patients with DM, arthritis, and respiratory involvement in ILD forms. For patients with DM and JDM, the most problematic signs and symptoms fall into two general categories: cutaneous and muscular symptoms. Physical skin symptoms include rash, itching, pain, and soreness, with rash (which may be itchy and / or painful) often being the first sign of DM (Cleland and Venzke, 2003). Common muscle complaints in patients with DM include fatigue, tiredness, muscle weakness, and decreased endurance (Okogbaa and Batiste, 2019). For patients with PM, proximal muscle weakness, sometimes accompanied by pain, is the primary clinical symptom. Patients have difficulty climbing stairs, rising from a sitting position, or raising their arms above their heads ( Lundberg et al., 2021 ).

[0008] As with other autoimmune disorders, the interplay between genetic and environmental factors is thought to result in the clinical phenotype of IIM. While the exact processes and triggers of the immune system are poorly understood, there is evidence that both adaptive and innate immune mechanisms, as well as non-immune mechanisms, are involved in various types of IIM (Lundberg et al., 2021). Literature data suggests that various risk factors, such as the presence of certain autoantibodies, result in different clinical characteristics, response to treatment, and outcomes. The pathogenesis of DM and JDM is multifactorial and complex. A strong genetic component has been described in DM and JDM. Data from genotyping studies suggest an association between major histocompatibility complex polymorphisms and the development of DM in adults and children. There is also evidence that certain HLA alleles are associated with autoantibody production, which correlates with the clinical phenotype (DeWane et al., 2020). There are multiple environmental factors that can trigger immune activation. Some of the triggers that have been described in patients with DM and JDM include UV radiation, viral infections, medications, and smoking. As with DM and JDM, there are genetic and environmental factors that contribute to the development of clinical PM.

[0009] Treating PM or DM is a challenging task, due in large part to its rarity, its multiple clinical phenotypes, and the fact that the disease affects multiple organ systems and is generally refractory to treatment (Bogdanov et al., 2018). There is no known cure for either PM or DM, and despite the significant morbidity and mortality associated with this condition, there is currently only one pharmacological treatment (Octagam 10% [IVIG]) recently approved for the treatment of DM in the United States and Europe based on a randomized controlled trial. The choice of treatment or the sequence in which various immunotherapeutic agents are used is often empirical rather than evidence-based, influenced by the clinician's experience, bias, and personal expectations regarding the efficacy / safety ratio of a given treatment (Dalakas, 2010).

[0010] The mainstay of treatment for either PM or DM is a combination of immunosuppressive drugs to treat inflammatory symptoms. First-line drug treatment for muscle diseases is typically systemic corticosteroids (e.g., high-dose systemic prednisolone) to address inflammation and suppress the immune system (Dalakas, 2010; Dalakas, 2011).

[0011] Other anti-inflammatory drugs, particularly azathioprine, methotrexate, MMF, hydroxychloroquine, cyclosporine, and cyclophosphamide, are used as next-line treatments in refractory cases or as steroid-sparing agents (Findlay et al., 2015). The preference for these drugs is empirical and not evidence-based, and their use as sole treatment appears to offer little benefit (Findlay et al., 2015). Furthermore, these agents are also associated with common and significant toxicities, including thrombocytopenia, anemia, leukopenia, and pancytopenia (azathioprine), liver and bone marrow toxicity (methotrexate, cyclosporine, cyclophosphamide), kidney toxicity (cyclosporine), and gastrointestinal symptoms and leukopenia (MMF) (Dalakas, 2010). Despite their empirical use, evidence from randomized clinical trials for these treatments remains limited (Fasano et al., 2016). The complications of long-term corticosteroid and other immunosuppressive therapies are well documented (Dalakas, 2010; Bradford Rice et al., 2016; Oray et al., 2016; Rice et al., 2017). Azathioprine and several corticosteroid products (e.g., prednisolone, betamethasone, dexamethasone) are nationally approved in several European countries for the treatment of IIM and a range of related conditions. A generic version of azathioprine (Jayempi) was recently approved in Europe by centralized procedure for the treatment of several inflammatory conditions, including DM (but not PM), in patients who are intolerant to glucocorticosteroids or have an inadequate therapeutic response despite treatment with high-dose glucocorticosteroids (EMA, 2021). However, it should be clear that these approvals were based primarily on empirical data and reports in the literature (as discussed above), rather than on data obtained from controlled clinical trials. These treatments are typically used in combination with other drugs and procedures.

[0012] After positive outcomes in several small case studies, other approaches, including tacrolimus (a broad-spectrum immunosuppressant developed for transplantation), IVIg, and rituximab (an anti-CD20 mAb), have emerged as potential treatments (Fasano et al., 2016). However, the safety profiles of these drugs are also difficult to manage in clinical settings (Dalakas, 2010; Fasano et al., 2016). Octagam 10% (IVIg) was recently approved in Europe (first approved in Germany in May 2021) through a decentralized review process for the treatment of adults with active DM who are treated with immunosuppressants, including corticosteroids, or who are intolerant of or have contraindications to these medications (Octapharma press release, June 2021), and in the United States (July 2021) for the treatment of DM in adults. These approvals were based on results from a single phase 3 randomized controlled trial of IVIg for dermatomyositis (ProDERM trial, NCT02728752). It should be noted that Octagam is associated with several safety risks, including thrombosis, renal dysfunction, and acute renal failure, and that its podiatry, requiring administration by IV infusion of two to five high doses over several consecutive days each month, imposes an additional burden on patients. Octagam is not currently approved for PMDA.

[0013] First-line treatment for patients with JDM is typically corticosteroids and / or methotrexate. Second-line treatments include IVIg, rituximab, cyclosporine, azathioprine, tacrolimus, and mycophenolate mofetil. Third-line treatments include stem cell transplantation or cyclophosphamide (Robinson and Reed, 2011). Octagam is not currently approved for the treatment of JDM.

[0014] Furthermore, even with current treatments, a significant number of patients remain treatment-refractory. Currently used treatments are also associated with significant safety issues. Recently, Octagam (10% IVIg) was approved in the United States and Europe for the treatment of adult DM, but not for JDM or PM. Octagam is associated with several risks (e.g., thrombosis, renal dysfunction, acute renal failure) and has a posology that requires monthly infusions over several consecutive days. Therefore, IIM (DM, PM, and JDM) represent a significant unmet need for effective and safe therapies, and therefore, there is a need for additional treatments for idiopathic inflammatory myopathies. Summary of the Invention [Problem to be solved by the invention]

[0015] The present invention provides a method of treating a patient having a condition associated with abnormal levels of IFNβ expression, comprising administering an anti-IFNβ antibody. The present invention further provides a method of treating a patient having idiopathic inflammatory myopathy (IIM), comprising administering an anti-IFNβ antibody. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by embodiment (E) below. [Means for solving the problem]

[0016] E1. In a first embodiment, the invention relates to a method for treating a patient having one or more conditions associated with IFNβ expression in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0017] E2. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0018] E3. In another embodiment, the invention relates to a method for treating one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, and cutaneous lupus in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0019] E4. In a first embodiment, the present invention relates to a method for treating IIM in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least 4 weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least 4 weeks.

[0020] E5. In another embodiment, the invention relates to a method for treating dermatomyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of dermatomyositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0021] E6. In another embodiment, the invention relates to a method for treating polymyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of polymyositis by at least 4 weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least 4 weeks.

[0022] E7. In another embodiment, the invention relates to a method for treating inclusion body myositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of inclusion body myositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0023] E8. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of SLE and cutaneous lupus, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least four weeks.

[0024] E9. In another embodiment, the invention relates to a method for treating a patient having one or more conditions associated with IFNβ expression in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0025] E10. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0026] E11. In another embodiment, the invention relates to a method for treating one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, and cutaneous lupus in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0027] E12. In a first embodiment, the present invention relates to a method for treating IIM in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0028] E13. In another embodiment, the invention relates to a method for treating dermatomyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of dermatomyositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0029] E14. In another embodiment, the invention relates to a method for treating polymyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of polymyositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0030] E15. In another embodiment, the invention relates to a method for treating inclusion body myositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of inclusion body myositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0031] E16. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of SLE and cutaneous lupus, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

[0032] E17. In another embodiment, the invention relates to a method for treating a patient having one or more conditions associated with IFNβ expression in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0033] E18. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses that are at least two weeks apart from each other.

[0034] E19. In another embodiment, the invention relates to a method for treating one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, and cutaneous lupus in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0035] E20. In a first embodiment, the present invention relates to a method for treating IIM in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0036] E21. In another embodiment, the invention relates to a method for treating dermatomyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of dermatomyositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0037] E22. In another embodiment, the invention relates to a method for treating polymyositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of polymyositis by at least 4 weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least 2 weeks.

[0038] E23. In another embodiment, the invention relates to a method for treating inclusion body myositis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of inclusion body myositis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0039] E24. In another embodiment, the invention relates to a method for treating a patient having one or more conditions selected from the group consisting of SLE and cutaneous lupus, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0040] E25. The method of E1 to E24, wherein one or more of the individual doses are administered at least one month apart.

[0041] E26. The method of E1 to E25, wherein one or more of the individual doses are administered at least 8 weeks apart.

[0042] E27. The method of E1 to E26, wherein one or more of the individual doses are administered at least two months apart.

[0043] E28. The method of E1 to E27, wherein one or more of the individual doses is in an amount of 25 mg to 1000 mg.

[0044] E29. The method of E1 to E28, wherein one or more of the individual doses is in an amount of 150 mg to 600 mg.

[0045] E30. The method of E1 to E29, wherein one or more of the individual doses is an amount selected from the group consisting of 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg.

[0046] E31. The method of E1 to E30, wherein one or more of the individual doses is an amount within a range selected from the group consisting of: lower limits of 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, and 600 mg.

[0047] E32. The method of E1 to E31, wherein one or more of the individual doses is an amount within a range selected from the group consisting of upper limits of 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg.

[0048] E33. The method of E1 to E32, wherein one or more of the individual doses is an amount within a range having a lower limit selected from the group consisting of 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, and 600 mg, and an upper limit selected from the group consisting of 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg.

[0049] E34. The method of E1 to E33, wherein one or more of the individual doses is in an amount of at least 140 mg.

[0050] E35. The method of E1 to E34, wherein one or more of the individual doses is in an amount of at least 150 mg.

[0051] E36. The method of E1 to E35, wherein one or more of the individual doses is in an amount of at least 200 mg.

[0052] E37. The method of E1 to E36, wherein one or more of the individual doses is in an amount of at least 250 mg.

[0053] E38. The method of E1 to E37, wherein one or more of the individual doses is in an amount of at least 300 mg.

[0054] E39. The method of E1 to E38, wherein one or more of the individual doses is in an amount of at least 350 mg, or at least 400 mg.

[0055] E40. The method of E1 to E39, wherein one or more of the individual doses is in an amount of at least 450 mg.

[0056] E41. The method of E1 to E40, wherein one or more of the individual doses is in an amount of at least 500 mg.

[0057] E42. The method of E1 to E41, wherein one or more of the individual doses is in an amount of at least 550 mg.

[0058] E43. The method of E1 to E42, wherein one or more of the individual doses is in an amount of at least 600 mg.

[0059] E44. The method of E1 to E43, wherein one or more of the individual doses is in an amount of 140 mg.

[0060] E45. The method of E1 to E44, wherein one or more of the individual doses is in an amount of 150 mg.

[0061] E46. The method of E1 to E45, wherein one or more of the individual doses is in an amount of 200 mg.

[0062] E47. The method of E1 to E46, wherein one or more of the individual doses is in an amount of 250 mg.

[0063] E48. The method of E1 to E47, wherein one or more of the individual doses is in an amount of 300 mg.

[0064] E49. The method of E1 to E48, wherein one or more of the individual doses is in an amount of 350 mg.

[0065] E50. The method of E1 to E49, wherein one or more of the individual doses is in an amount of 450 mg.

[0066] E51. The method of E1 to E50, wherein one or more of the individual doses is in an amount of 500 mg.

[0067] E52. The method of E1 to E51, wherein one or more of the individual doses is in an amount of 550 mg.

[0068] E53. The method of E1 to E52, wherein one or more of the individual doses is in an amount of 600 mg.

[0069] E54. The method of E1 to E53, wherein one or more of the individual doses is in an amount of 900 mg.

[0070] E55. The method of any one of E1 to E54, wherein the majority of the individual doses are the same amount.

[0071] E56. The method of E1 to E53, wherein each dose is the same amount.

[0072] E57. The method of any one of E1 to E56, wherein one or more of the individual doses is by intravenous injection.

[0073] E58. The method of any one of E1 to E57, wherein the multiple individual doses are by intravenous injection.

[0074] E59. The method of any one of E1 to E58, wherein the multiple individual doses are by intravenous injection and in an amount of at least 150 mg.

[0075] E60. The method of any one of E1 to E59, wherein the multiple individual doses are by intravenous injection and in an amount of at least 450 mg.

[0076] E61. The method of any one of E1 to E60, wherein the multiple individual doses are by intravenous injection and in an amount of at least 600 mg.

[0077] E62. The method of any one of E1 to E61, wherein the multiple individual doses are by intravenous injection and are separated from each other by at least 4 weeks.

[0078] E63. The method of any one of E1 to E62, wherein the multiple individual doses are by intravenous injection, are in amounts of at least 150 mg, and are separated from each other by at least 4 weeks.

[0079] E64. The method of any one of E1 to E63, wherein the multiple individual doses are by intravenous injection, are in amounts of at least 450 mg, and are separated from each other by at least 4 weeks.

[0080] E65. The method of any one of E1 to E64, wherein the multiple individual doses are by intravenous injection, are in amounts of at least 600 mg, and are separated from each other by at least 4 weeks.

[0081] E66. The method of any one of E1 to E56, wherein one or more of the individual doses is by subcutaneous injection.

[0082] E67. The method of E67, wherein the multiple individual doses are by subcutaneous injection.

[0083] E68. The method of any one of E66 to E67, wherein the multiple individual doses are by subcutaneous injection and in an amount of at least 150 mg.

[0084] E69. The method of any one of E66 to E68, wherein the multiple individual doses are by subcutaneous injection and in an amount of at least 450 mg.

[0085] E70. The method of any one of E66 to E69, wherein the plurality of individual doses is by subcutaneous injection and in an amount of at least 600 mg.

[0086] E71. The method of any one of E66 to E70, wherein the multiple individual doses are by subcutaneous injection and are separated from each other by at least one week.

[0087] E72. The method of any one of E66 to E71, wherein the multiple individual doses are by subcutaneous injection, are in amounts of at least 150 mg, and are separated from each other by at least 1 week.

[0088] E73. The method of any one of E66 to E72, wherein the multiple individual doses are by subcutaneous injection, are in an amount of at least 450 mg, and are separated from each other by at least 1 week.

[0089] E74. The method of any one of E66 to E73, wherein the multiple individual doses are by subcutaneous injection, are in an amount of at least 600 mg, and are separated from each other by at least 1 week.

[0090] E75. The method of E1 to E74, wherein the dosing regimen is continued for at least 4 weeks.

[0091] E76. The method of E1 to E75, wherein the dosing regimen is continued for at least one month.

[0092] E77. The method of E1 to E76, wherein the dosing regimen is continued for at least 8 weeks.

[0093] E78. The method of E1 to E77, wherein the dosing regimen is continued for at least 2 months.

[0094] E79. The method of E1 to E78, wherein the dosing regimen is continued for at least 12 weeks.

[0095] E80. The method of E1 to E79, wherein the dosing regimen is continued for at least 3 months.

[0096] E81. The method of E1 to E80, wherein the dosing regimen is continued for at least 16 weeks.

[0097] E82. The method of E1 to E81, wherein the dosing regimen is continued for at least 4 months.

[0098] E83. The method of E1 to E82, wherein the dosing regimen is continued for at least 20 weeks.

[0099] E84. The method of E1 to E83, wherein the dosing regimen is continued for at least 5 months.

[0100] E85. The method of E1 to E84, wherein the dosing regimen is continued for at least 24 weeks.

[0101] E86. The method of E1 to E85, wherein the dosing regimen is continued for at least 6 months.

[0102] E87. The method of E1 to E86, wherein the dosing regimen is continued for at least 26 weeks.

[0103] E88. The method of any one of E1 to E87, wherein the improvement in signs or symptoms is characterized by a clinical response.

[0104] E89. The method of any one of E1 to E88, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences an improvement in signs and symptoms of one or more selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, juvenile dermatomyositis, SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma, characterized by a clinical response.

[0105] E90. The method of any one of E1 to E89, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences an improvement in signs and symptoms of one or more selected from the group consisting of SLE, cutaneous lupus, and psoriasis, characterized by a clinical response.

[0106] E91. The method of any one of E1 to E89, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms of one or more selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, and juvenile dermatomyositis.

[0107] E92. The method of any one of E1 to E91, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences an improvement in IIM, characterized by a clinical response.

[0108] E93. The method of any one of E1 to E92, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in the dermatomyositis, characterized by a clinical response.

[0109] E94. The method of any one of E1 to E93, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in polymyositis, characterized by a clinical response.

[0110] E95. The method of any one of E1 to E94, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in the inclusion body myositis, characterized by a clinical response.

[0111] E96. The method of any one of E1 to E95, wherein at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in the juvenile dermatomyositis, characterized by a clinical response.

[0112] E97. The method of any one of E1 to E76, wherein the clinical response is measured by assessment of skin lesions.

[0113] E98. The method of E88 to E97, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of greater than 0.

[0114] E99. The method of E88 to E98, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of at least 5.

[0115] E100. The method of E88 to E99, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of at least 7.

[0116] E101. The method of E88 to 100, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of at least 9.

[0117] E102. The method of E88 to E101, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of at least 15.

[0118] E103. The method of E88 to E102, wherein the clinical response can be characterized by a change from baseline in Manual Muscle Testing (MMT-8) score of at least 20.

[0119] E104. The method of E88 to 103, wherein the clinical response may be characterized by an improvement in Total Improvement Score (TIS) greater than 0.

[0120] E105. The method of E88 to E104, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 20.

[0121] E106. The method of E88 to E105, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 25.

[0122] E107. The method of E88 to E106, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 30.

[0123] E108. The method of E88 to E107, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 35.

[0124] E109. The method of E88 to E108, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 40.

[0125] E110. The method of E88 to E109, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 45.

[0126] E111. The method of E88 to E110, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 50.

[0127] E112. The method of E88 to E111, wherein the clinical response can be characterized by an improvement in Total Improvement Score (TIS) of at least 55.

[0128] E113. The method of E88 to E112, wherein the clinical response can be characterized by a change from baseline in patient global assessment score below 0.

[0129] E114. The method of E88 to E113, wherein the clinical response can be characterized by a change from baseline in patient global assessment score of at least -1 on a 10-centimeter visual analog scale (VAS).

[0130] E115. The method of E88 to E114, wherein the clinical response can be characterized by a change from baseline in patient global assessment score of at least -2 on a 10-centimeter VAS.

[0131] E116. The method of E88 to E115, wherein the clinical response can be characterized by a change from baseline in patient global assessment score of at least -3 on a 10-centimeter VAS.

[0132] E117. The method of E88 to E116, wherein the clinical response can be characterized by a change from baseline in patient global assessment score of at least -4 on a 10-centimeter VAS.

[0133] E118. The method of E88 to E117, wherein the clinical response can be characterized by a change from baseline in patient global assessment score of at least -5 on a 10-centimeter VAS.

[0134] E119. The method of E88 to E118, wherein the clinical response can be characterized by an improvement in absolute value of the muscle enzyme creatine kinase.

[0135] E120. The method of E88 to E119, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -75 U / L.

[0136] E121. The method of E88 to E120, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -100 U / L.

[0137] E122. The method of E88 to E121, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -125 U / L.

[0138] E123. The method of E88 to E122, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -150 U / L.

[0139] E124. The method of E88 to E123, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -175 U / L.

[0140] E125. The method of E88 to E124, wherein the clinical response can be characterized by an improvement in absolute muscle enzyme creatine kinase of at least -185 U / L.

[0141] E126. The method of E88 to E125, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) greater than 0.

[0142] E127. The method of E88 to E126, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) of at least -5.

[0143] E128. The method of E88 to E127, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) of at least -10.

[0144] E129. The method of E88 to E128, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) of at least -12.

[0145] E130. The method of E88 to E129, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) of at least -14.

[0146] E131. The method of E88 to E130, wherein the clinical response can be characterized by a change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) of at least -20.

[0147] E132. The method of any one of E1 to E131, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 4 weeks.

[0148] E133. The method of any one of E1 to 132, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 6 weeks.

[0149] E134. The method of any one of E1 to E133, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 8 weeks.

[0150] E135. The method of any one of E1 to E134, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 12 weeks.

[0151] E136. The method of any one of E1 to E135, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 16 weeks.

[0152] E137. The method of any one of E1 to E136, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 20 weeks.

[0153] E138. The method of any one of E1 to E137, wherein at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms that is maintained for at least an additional 24 weeks.

[0154] E139. The method of E1 to E138, wherein improvement in signs and symptoms is experienced at least 8 weeks after initiation of the medication regimen.

[0155] E140. The method of E1 to E139, wherein improvement in signs and symptoms is experienced at least 10 weeks after initiation of the medication regimen.

[0156] E141. The method of E1 to E140, wherein improvement in signs and symptoms is experienced at least 12 weeks after initiation of the medication regimen.

[0157] E142. The method of E1 to E141, wherein improvement in signs and symptoms is experienced at least 16 weeks after initiation of the medication regimen.

[0158] E143. The method of E1 to E142, wherein improvement in signs and symptoms is experienced at least 20 weeks after initiation of the medication regimen.

[0159] E144. The method of E1 to E143, wherein improvement in signs and symptoms is experienced at least 24 weeks after initiation of the medication regimen.

[0160] E145. The method of any of E1 to E144, wherein the patient is treated with at least one other medication concurrently with the dosing regimen with anti-IFNβ.

[0161] E146. The method of any of E1 to E145, wherein the patient has been previously treated with at least one other medication.

[0162] E147. The method of E125 to E146, wherein the at least one other medication is selected from the group consisting of corticosteroids, IVIG, and immunomodulatory and immunosuppressive drugs.

[0163] E148. The method of E147, wherein the immunomodulatory and immunosuppressive agents are selected from the group consisting of hydroxychloroquine, azathioprine, mycophenolate mofetil, and methotrexate.

[0164] E149. The method of E1 to E148, wherein the patient exhibits a clinical response after 24 weeks of treatment.

[0165] E150. The method of E1 to E149, wherein the patient exhibits a clinical response after 20 weeks of treatment.

[0166] E151. The method of E1 to E150, wherein the patient exhibits a clinical response after 16 weeks of treatment.

[0167] E152. The method of E1 to E151, wherein the patient exhibits a clinical response after 12 weeks of treatment.

[0168] E153. The method of E1 to E152, wherein the patient exhibits a clinical response 8 weeks after treatment.

[0169] E154. The method of E1 to E153, wherein the patient exhibits a clinical response 4 weeks after treatment.

[0170] E155. The method of E1 to E154, wherein the anti-IFNβ antibody comprises three CDRs derived from a variable heavy chain region having the sequence set forth in SEQ ID NO:3, and three CDRs derived from a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0171] E156. The method of E1 to E155, wherein the anti-IFNβ antibody comprises an HCDR1 having the sequence set forth in SEQ ID NO: 5, an HCDR2 having the sequence set forth in SEQ ID NO: 6, an HCDR3 having the sequence set forth in SEQ ID NO: 7, an LCDR1 having the sequence set forth in SEQ ID NO: 8, an LCDR2 having the sequence set forth in SEQ ID NO: 9, and an LCDR3 having the sequence set forth in SEQ ID NO: 10.

[0172] E157. The method of E1 to E156, wherein the anti-IFNβ antibody comprises a variable heavy chain region having the sequence set forth in SEQ ID NO:3, and a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0173] E158. The method of E1 to E157, wherein the anti-IFNβ antibody comprises a heavy chain having the sequence set forth in SEQ ID NO: 1 and a light chain having the sequence set forth in SEQ ID NO: 2, and the C-terminal lysine (K) of the heavy chain amino acid sequence of SEQ ID NO: 1 is optional.

[0174] E159. The method of E1 to E158, wherein the anti-IFNβ antibody comprises a VH encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VH having ATCC accession number PTA-122727, and a VL encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VL having ATCC accession number PTA-122726.

[0175] E160. The method of any one of E1 to E159, wherein the antibody comprises a VH sequence encoded by an insert in a plasmid deposited with the ATCC having ATCC Accession No. PTA-122727.

[0176] E161. The method of any one of E1 to E160, wherein the antibody comprises a VL sequence encoded by an insert in a plasmid deposited with the ATCC having ATCC Accession No. PTA-122726.

[0177] E162. The method of E1 to E161, wherein the anti-IFNβ antibody competes for binding with an anti-IFNβ antibody comprising a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0178] E163. The method of E1 to E162, wherein the anti-IFNβ antibody competes for binding with an antibody comprising a VH encoded by the nucleic acid sequence of an insert in the vector deposited as CTI-AF1-VH having ATCC accession number PTA-122727 and a VL encoded by the nucleic acid sequence of an insert in the vector deposited as CTI-AF1-VL having ATCC accession number PTA-122726.

[0179] E164. The method of any one of E1 to E162, wherein the JDM patient is one or more of: at least 12 years old, weighs at least 30 kg, or weighs at least 40 kg.

[0180] E165. Use of an anti-IFNβ antibody for the preparation of a medicament for the method of treatment according to any of E1 to E164.

[0181] E166. An anti-IFNβ antibody for use in the method according to any one of E1 to E165.

[0182] E167. Use of an anti-IFN-beta antibody in the preparation of a medicament for treating a patient according to the method of any one of E1 to E166.

[0183] E168. Aqueous preparations, anti-IFNβ antibody at a concentration of 25 mg / mL to 200 mg / mL; histidine or His-HCl at a concentration of 10–50 mM; arginine or NaCl in amounts of 20-150 mM, containing sucrose or trehalose in an amount of 20 mg / ml to 85 mg / ml; pH ranges from pH 5.0 to pH 6.5. Aqueous formulation.

[0184] E169. The formulation according to E168, further comprising a chelating agent.

[0185] E170. The formulation according to E169, wherein the chelating agent is present in an amount of 0.01 to 0.1 mg / ml.

[0186] E171. The formulation of E170, wherein the chelating agent is present in an amount of 0.02 to 0.08 mg / ml.

[0187] E172. The formulation of E171, wherein the chelating agent is present in an amount of 0.05 mg / ml.

[0188] E173. A formulation according to E169 to E172, wherein the chelating agent is EDTA.

[0189] E174. A formulation according to E169 to E173, wherein the chelating agent is EDTA and is present in an amount of 0.05 mg / ml.

[0190] E175. The formulation according to E168-E174, further comprising a surfactant.

[0191] E176. A formulation according to E174 to E175, wherein the surfactant is present in an amount of 0.05 to 0.5 mg / ml.

[0192] E177. A formulation according to E174 to E176, wherein the surfactant is present in an amount of 0.1 to 0.3 mg / ml.

[0193] E178. A formulation according to E174 to E177, wherein the surfactant is present in an amount of 0.2 mg / ml.

[0194] E179. The formulation according to E174 to E178, wherein the surfactant is PS80.

[0195] E180. A formulation according to E174 to E179, wherein the surfactant is PS80 and is present in an amount of 0.2 mg / ml.

[0196] E181. The formulation of E168 to E180, wherein histidine or His-HCL is present in an amount of 2 mM to 50 mM.

[0197] E182. The formulation of E168 to E181, wherein histidine or His-HCl is present in an amount of 5 mM to 30 mM.

[0198] E183. The formulation of E168 to E182, wherein histidine or His-HCl is present in an amount of 10 mM to 30 mM.

[0199] E184. The formulation of E168 to E183, wherein histidine or His-HCL is present in an amount of 20 mM.

[0200] E185. A formulation according to E168 to E184, comprising histidine.

[0201] E186. The formulation according to E168 to E185, comprising histidine in an amount of 20 mM.

[0202] E187. The formulation according to E168 to E186, wherein arginine or NaCl is present in an amount of 50 mM to 150 mM.

[0203] E188. The formulation according to E168 to E187, wherein arginine or NaCl is present in an amount of 50 mM to 100 mM.

[0204] E189. The formulation of E168 to E188, wherein arginine or NaCl is present in an amount of 50 mM.

[0205] E190. A formulation according to E168 to E189, comprising arginine.

[0206] E191. The formulation according to E168 to E190, comprising arginine in an amount of 50 mM.

[0207] E192. A formulation according to E168 to E191, wherein the sucrose or trehalose is present in an amount of 25 to 75 mg.ml.

[0208] E193. The formulation according to E168 to E192, wherein the sucrose or trehalose is present in an amount of 50 to 75 mg.ml.

[0209] E194. The formulation of E168 to E193, wherein the sucrose or trehalose is present in an amount of 50 mg.ml.

[0210] E195. A formulation according to E168 to E194, comprising sucrose.

[0211] E196. A formulation according to E168 to E195, comprising sucrose in an amount of 50 mg / ml.

[0212] E197. The formulation according to E168 to E196, wherein the pH is 5.5 to 6.0.

[0213] E198. The formulation according to E168 to E197, wherein the pH is 5.8.

[0214] E199. The formulation of E168 to E198, wherein the anti-IFNβ antibody is present in an amount of at least 50 mg.ml.

[0215] E200. The formulation of E168 to E199, wherein the anti-IFNβ antibody is present in an amount of at least 60 mg.ml.

[0216] E201. The formulation of E168 to E200, wherein the anti-IFNβ antibody is present in an amount of 60 mg.ml.

[0217] E202. The formulation of E168 to E201, wherein the anti-IFNβ antibody is present in an amount of at least 80 mg.ml.

[0218] E203. The formulation of E168 to E202, wherein the anti-IFNβ antibody is present in an amount of at least 100 mg.ml. or at least 120 mg.mL.

[0219] E204. The formulation of E168 to E203, wherein the anti-IFNβ antibody is present in an amount of about 141-154 mg.mL.

[0220] E205. The formulation of E168 to E204, wherein the anti-IFNβ antibody is present in an amount of at least 150 mg.ml.

[0221] E206. The formulation according to E168 to E205, wherein the anti-IFNβ antibody is present in an amount of 150 mg.ml.

[0222] E207. The formulation of E168 to E206, wherein the anti-IFNβ antibody is as described in any one of E155 to E163.

[0223] E208. 60 mg / ml of an anti-IFNβ according to any one of E155 to E163; 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / ml EDTA, Contains 0.2 mg / ml polysorbate 80, pH 5.8 Preparations according to E168 to E207.

[0224] E209. 150 mg / ml of an anti-IFNβ according to any one of E155 to E163; 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / ml EDTA, Contains 0.2 mg / ml polysorbate 80, pH 5.8 Preparations according to E168 to E207.

[0225] E210. 141 to 154 mg / ml of the anti-IFNβ according to any one of E155 to E163; 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / ml EDTA, Contains 0.2 mg / ml polysorbate 80, pH 5.8 Preparations according to E168 to E207.

[0226] E211. A formulation according to any one of E168 to E210, having a viscosity of less than 20 centipoise.

[0227] E212. A formulation according to any one of E168 to E211, having a viscosity of less than 15 centipoise.

[0228] E213. The formulation of any one of E168 to E212, having an osmolality of less than 500 mOsm.

[0229] E214. The formulation of any one of E168 to E213, having an osmolality of less than 400 mOsm.

[0230] E215. A formulation according to any one of E168 to E214 for use in a method according to any one of E1 to E143, or for use according to E144 or E146.

[0231] E216. The method of any one of E1 to E164, wherein the antibody is formulated in a formulation of any one of E168 to E215.

[0232] E217. The method of any one of E1 to E164 comprising administering to a subject a therapeutically effective amount of the aqueous formulation of any one of E168 to E215. [Brief explanation of the drawings]

[0233] [Figure 1] Stage 1 (Planned Sample Size). Participants with skin lesions (CDASI-activity ≥ 14 at screening) who had failed at least one standard of care systemic treatment (e.g., corticosteroids) were randomized in a 2:1 ratio to receive 600 mg of PF-06823859 or placebo. Study drug or placebo administration occurred on days 1, 4, and 8. The primary endpoint (CFB CDASI-A) was assessed at week 12. [Figure 2] Stage 2 (Planned Sample Size). Participants with skin involvement (CDASI-activity ≥ 14 at screening) were randomized in a 5:11:4 ratio to receive 600 mg of PF-06823859, 150 mg of PF-06823859, or placebo. Study drug or placebo administration occurred on days 1, 4, and 8 of the study. The primary endpoint (CFB CDASI-A) was assessed at week 12. [Figure 3]Accrual Stage 2 (Planned Sample Size). Accrual Stage 2 employed a fixed-sequence design, allowing all study participants the opportunity to receive active medication during the treatment period. Participants were randomized in a 5:11:2:2 ratio to one of the following sequences: 600 mg PF-06823859 followed by placebo, 150 mg PF-06823859 followed by placebo, placebo followed by 600 mg PF-06823859, or placebo followed by 150 mg PF-06823859. Administration of study medication or placebo (determined by treatment sequence) occurred on Day 1, Week 4, Week 8, Week 12, Week 16, and Week 20. The primary endpoint (CFB CDASI-A) was assessed at Week 12 of Accrual Stage 2. After the treatment period ended at 24 weeks, participants then entered a 4-month follow-up period or were transferred to the long-term extension study C0251008. [Figure 4]Stage 3 (Planned Sample Size). Stage 3 also employed a fixed-sequence design, randomizing primarily muscle pathology participants 1:1 to one of the following sequences: 600 mg PF-06823859 followed by placebo, or placebo followed by 600 mg PF-06823859, with treatment switching at week 12. Muscle pathology inclusion criteria required subjects to meet one of the following two criteria: (1) MMT-8 ≤ 136 / 150 and PhGA (VAS ≥ 3 cm on a 0-10 cm scale), or (2) the sum of PhGA, PtGA, and extramuscular global assessment ≥ 10 cm (each using a 0-10 cm VAS scale) and failure of at least two adequate courses of immunosuppressants or immunomodulators, including IVIG. Concomitant medications included stable doses of immunosuppressants and immunomodulators, including intravenous immunoglobulin (IVIG). Study drug or placebo administration (determined by treatment sequence) occurred on Day 1, Weeks 4, 8, 12, 16, and 20. Secondary endpoints (TIS) were assessed longitudinally at Weeks 4, 8, and 12 (Week 12 was the critical time point). Other secondary muscle-related endpoints included MMT-8 and PtGA of myositis. After Week 12, the sequence was switched to the other treatment. After the treatment period ended at Week 24, participants entered a 4-month follow-up period or entered the long-term extension study C0251008. [Figure 5] Mean absolute TIS values ​​and difference from placebo (MMRM, weeks 4 to 12). [Figure 6] Mean change from baseline in MMT-8 (LANCOVA, weeks 4 to 12). [Figure 7] Change from baseline in mean patient global assessment of myositis (LANCOVA) using a 10 cm (i.e., 100 mm) VAS ranging from 0 to 100. [Figure 8] Mean change from baseline in CK (U / L) (LANCOVA, weeks 1 to 12). [Figure 9]Estimated CFB of mean CDASI activity scores (LANCOVA-P, baseline-week 12, FAS1 and pooled FAS of the skin cohort (PFASS, which includes the first 12 weeks of data from subjects in Stage 1, Stage 2, and corrected Stage 2). [Figure 10] Strength and significance of the 10-gene type 1 IFN signature across related diseases, indicating further opportunities beyond dermatomyositis ("DM"). [Figure 11] Illustration of the effect of Pfizer's anti-IFNB molecule in a dermatomyositis trial divided by dose arms. Patients within each dose arm have comparable type 1 IFN signatures at baseline (baseline: circles). After treatment, the type 1 IFN signature in placebo-treated patients increased slightly, while the signature of the anti-IFNB molecule decreased in both dose arms (treatment: triangles). [Figure 12] Figure 12A: Effect of pH on anti-IFNβ viscosity at different concentrations. Figure 12B: Effect of formulation on anti-IFNβ viscosity at different concentrations. [Figure 13] Effect of arginine concentration on anti-IFNβ viscosity. [Figure 14] Effect of sodium chloride on anti-IFNβ viscosity. [Figure 15] Visual predictability assessment of the final pharmacokinetic profile. Speckled represents the median at each binning time point, with the 5th and 95th percentiles hatched. The solid line is the observed value, the ribbon is the simulated 90% distribution, and the dashed line is the simulated median. [Figure 16] Visual predictive assessment of IFNβ in the final PKPD model. Speckled represents the median at each binning time point, with the 5th and 95th percentiles hatched. Solid lines are observed, ribbons are the simulated 90% distribution, and dashed lines are the simulated median. Observations are black dots. [Figure 17]Simulation of pharmacokinetics of doses followed for 52 weeks. PF-06823859 median plasma concentrations are shown as solid black lines, with the 90% prediction interval for the median shown as dashed lines. Predicted drug-IFNβ complex concentrations are shown in gray below and are in pg / mL. KSS is shown as a dotted gray line and is in ng / mL. [Figure 18] Simulation of biomarkers after 52 weeks of dosing. All solid lines represent medians, and dashed lines are 90% prediction intervals for the medians. For IFNβ, the horizontal dashed line indicates the LLOQ, and the dotted line indicates the median boundary of the simulated 90% distribution of IFNβ. [Figure 19] Summary of dose and frequency options and simulated biomarker responses. For Figures 19(A), 19(B), and 19(C), the frequency of dosing (if greater than 1) is fixed at every 4 weeks. For Figures 19(D), 19(E), and 19(F), the number of doses is fixed at 3. Points represent median endpoints, and error bars represent 90% prediction intervals for the median. Lines simply connect the points and are not smoothed or imputed. [Figure 20] Simulated percent of adolescent subjects with exposure ratios greater than 2 for various weight-based dosing thresholds. At each cutoff, subjects below the index weight receive a 9 mg / kg dose, and those above receive a 600 mg dose. The median (black, solid line) percent of subjects at weight-based dosing cutoffs of 30 to 70 kg and their 90% prediction intervals (gray ribbons) for the median are shown. The vertical solid line and corresponding label indicate where the median crosses the 10% threshold, and the 90% prediction interval indicates where the upper and lower estimates cross the 10% threshold. [Figure 21]PK simulation of trough matching between SC dosing regimens and reference IV administration. The x-axis represents time after dosing in weeks, and the y-axis represents geometric mean concentration (ng / mL). Dosing regimens are indicated by dotted lines (600 mg IV Q4W), dot-dashed lines (750 mg SC Q4W), dashed lines (300 mg SC Q2W), and thin-dashed lines (150 mg SC QW). Abbreviations: IV, intravenous; ml, milliliters; ng, nanograms; QW, weekly; Q2W, q2w; Q4W, q4w; SC, subcutaneous. [Figure 22] PK simulation to match AUC / Cave between SC dosing regimens and reference IV administration. The X-axis represents time after dosing in weeks, and the Y-axis represents geometric mean concentration (ng / mL). Dosing regimens are indicated by dot-dashed lines (600 mg IV Q4W), dotted lines (900 mg SC Q4W), and dashed lines (225 mg SC QW). Abbreviations: IV, intravenous; ml, milliliters; ng, nanograms; QW, weekly; Q4W, every 4 weeks; SC, subcutaneous. DETAILED DESCRIPTION OF THE INVENTION

[0234] The Stage 2 (dose ranging in skin-predominant DM) and Stage 3 placebo-controlled 12-week (muscle-predominant DM) data indicated surprisingly high levels of efficacy. Notably, data from the muscle cohort of the Phase 2 study demonstrated efficacy that supported the efficacy data for skin conditions. Thus, this study demonstrates the ability of an IFNβ neutralizing antibody to provide effective treatment for DM / JDM and PM, and the modeling data provided herein supports efficacy at comparable doses for other indications, particularly SLE, cutaneous lupus, and psoriasis.

[0235] DM is characterized by B cell activation and antibody-mediated inflammation and tissue damage. Scientific literature on DM supports the hypothesis that IFNβ protein levels are elevated in DM blood and mRNA levels are increased in DM skin. Capelletti et al. showed that multiple components of the IFNα / β-mediated response were upregulated in muscle samples from myositis patients compared with controls (Capelletti et al., 2011). In their analysis, ISGI5, IFIT3, MX1, and IFIT1, which are regulators of the biological and therapeutic actions of IFNα and IFNβ, were the most highly expressed. In this study, IFN-mediated molecules were expressed in JDM, DM, and PM, with the highest expression observed in samples from JDM patients. Finally, IFNβ mRNA was upregulated, indicating that this IFN is likely responsible for the upregulation of the mentioned genes (Baechler et al., 2011).

[0236] Type 1 IFN-inducible genes are elevated in blood and muscle from patients with PM, but to a lesser extent than those seen in DM. Detection of IFNβ in PM samples is more limited, but may also be hampered by current assay sensitivity. When IFN expression levels in peripheral blood and muscle from patients with IIM were analyzed to determine their relationship to organ symptoms, the expression levels of IFNα and IFNβ in PM / DM patients were significantly higher than those in normal controls, and the increment correlated with disease activity and severity (Zu and Wang, 2011).

[0237] DM and JDM are sufficiently similar in terms of clinical presentation, pathophysiology, and histopathology that they can be evaluated for efficacy as a combined cohort, recognizing the rarity of DM and JDM.

[0238] As reported in the literature, when treatment appears to be effective, improvements in muscle function are observed within the first 6 months of treatment. An example of this is described by Ruperto et al. (Ruperto et al., 2016), who showed that more than 50% of JDM patients treated with either prednisone alone or prednisone in combination with methotrexate or cyclosporine achieved a 20% or greater improvement in three of six CSMs after 6 months of treatment. Similarly, the Proderm study showed that 78.7% of DM patients treated with Octagam achieved a minimal improvement in TIS after 16 weeks of treatment.

[0239] Given the early onset of skin efficacy (plateauing by week 12) and the clear muscle efficacy demonstrated by week 12 in participants receiving PF-06823859 in this study, assessment at week 24 (moderate improvement in TIS) will allow for observation of the maximum treatment effect of PF-06823859 on muscle and skin manifestations of disease. An exposure-response model relating IFNb inhibition to various clinical endpoints (including CDASI, SF-36, and TIS component scores) was developed, which predicts a strong TIS response that begins to plateau around week 12; therefore, a 24-week endpoint allows for consideration of the possibility of diminishing efficacy with a relatively short treatment period and, at the very least, a delay in achieving maximum efficacy in PM patients.

[0240] PM and DM patients share a common core characteristic: muscle weakness. Furthermore, type 1 IFN-inducible genes are elevated in the blood and muscle of patients with PM. Given the high level of target binding observed in DM patients and the clinical efficacy of a 600 mg Q4W dose, it is predicted that the same dose will achieve a similar level of efficacy in PM patients.

[0241] The TIS is a weighted measure of improvement from baseline in six Core Set Scales (CSM): 1) MDGA-VAS (Physician Global Assessment VAS, i.e., PhGA-VAS), 2) PtGA-VAS (Patient Global Assessment VAS), 3) MMT-8 (Manual Muscle Testing-8), 4) Muscle Enzymes, 5) MDAAT (Myositis Disease Activity Assessment Tool), and 6) HAQ-DI (Health Status Assessment Questionnaire-Disability Index, IMACS, Aggarwal et al., 2017). IMACS established consensus on a set of core domains and scales for the assessment of disease activity in patients with DM, JDM, and PM. The CSM has been endorsed by the ACR / EULAR and is recommended for use in clinical trials as an endpoint for assessing improvement in muscle symptoms in DM, JDM, and PM. This criteria set was approved by the ACR Board of Directors and the EULAR Executive Committee. The TIS has been approved by the ACR / EULAR / PRINTO Committee, which indicates that the criteria set has been quantitatively validated using patient data and has undergone validation based on independent datasets (Aggarwal et al., 2017). The TIS has also been evaluated by the PRINTO group as a CSM, demonstrating its usefulness in pediatric patients (Ruperto and Martini, 2011). Of note, as recommended by ACR / EULAR / PRINTO for the TIS, physical function in children, including adolescents, is measured by the CHAQ-Impairment Index, which assesses the impact of the disease on school activities and is derived from the HAQ-Impairment Index for adults, which assesses the impact of the disease on activities at work.

[0242] The TIS is the sum of the improvements reflected in each of the six CSMs (Table 1), but individual CSMs are weighted so that those considered more important contribute more to the final score. For example, a change in MMT score is weighted more heavily than a change in the most abnormal enzyme or HAQ (Aggarwal et al., 2017).

[0243] [Table 1-1]

[0244] [Table 1-2]

[0245] Considering the important clinical symptoms of DM, JDM, and PM in muscle and the significant impairment in patients' HRQoL, and given the validation of the CSM described above, the TIS is a valid assessment to assess the effect of treatment with PF-06823859 on the muscle symptoms of DM, JDM, and PM.

[0246] Rationale for Using Moderate Improvement in TIS as the Primary Endpoint The TIS provides a quantitative assessment of the degree of response to therapeutic intervention for each patient using a scale of 0 to 100. The thresholds for the TIS improvement categories are tabulated below in Table 2.

[0247] [Table 2]

[0248] A categorical outcome of response was validated as part of the 2016 ACR / EULAR Myositis Response Criteria for Adult DM / PM and JDM. Rider L. et al. described the Myositis Response Criteria and noted that, although validation was performed using limited data, they have several important advantages, including the feasibility of using the same definition of improvement (minimal, moderate, and large) across DM, PM, and JDM subtypes (Rider et al., 2018). Other advantages include the fact that they do not require a minimum level of myositis severity at baseline, and all levels of improvement in CSM contribute to a response.

[0249] CDASI-A The CDASI was designed to capture the extent of skin disease and was developed for use in clinical trials and longitudinal patient assessments (Anyanwu et al., 2015, British Journal of Dermatology, 173, pp. 969-974, incorporated herein by reference in its entirety). Disease lesions are measured at 15 different anatomical locations and graded using a) three activity scales (erythema, scaling, and erosion / ulceration), b) two damage scales (poikiloderma and calcinosis), and c) the presence and severity of Gottron plaques, periungual changes, and alopecia on the hands. The resulting activity and damage scores range from 0 to 100 and 0 to 32, respectively, with higher scores indicating greater disease severity.

[0250] The CDASI was developed with the intent of creating a valid and reliable measure of skin lesions in DM and has been shown to be a valid outcome measure in clinical trials. A separate study (Ahmed et al., 2020a; Ahmed et al., 2020b) in 103 patients with DM assessed the percentage change and actual change in the CDASI-A score required to achieve a meaningful improvement in QoL. The study concluded that a 40% change in the CDASI-A score between the first two visits represents a meaningful change in QoL in patients with a threshold CDASI-A range above 14 points (Ahmed et al., 2020a; Ahmed et al., 2020b). Additionally, in interviews conducted to gather information about content validity, three clinicians (n ​​= 3) reported a desired reduction of 6–10 points over a 12-week clinical trial, and two more (n = 2) reported a desired reduction of 5–8 points over a 24-week clinical trial. Previous studies have assessed the reliability of the CDASI in DM and JDM, confirming its reliability as an assessment tool (Tiao et al., 2017). CDASI development and validation work included characterizing disease severity and assessing the instrument's responsiveness to clinically meaningful change. Analysis demonstrated similar values ​​for disease severity characterization and clinical response. The authors concluded that a CDASI activity score of 19 or less characterizes mild disease, with a cutoff between 14 and 19. In a 2017 prospective evaluation of n = 42 patients with DM and n = 25 healthy participants, a type I IFN pathway signature biomarker in blood was found to be highly correlated with CDASI activity scores in DM. The correlation of serum IFNb with both the gene signature and the CDASI suggests that IFNb may signal disease activity in DM and may be a promising surrogate endpoint in clinical trials (Huard et al., 2017). Therefore, an improvement of at least 4 CDASI units may indicate significant clinical benefit.

[0251] MMT-8 The MMT-8 is a physical performance outcome assessment that objectively assesses muscle strength. The MMT-8 is a TIS CSM, and muscle function is a major factor affecting the lives of patients with DM, JDM, and PM, among others. The MMT-8 is included in the treatment guidelines for DM and PM as an IMACS CSM (Rider et al., 2010, incorporated herein by reference in its entirety) and for JDM as a CSM by IMACS and PRINTO (Rider et al., 2018, incorporated herein by reference in its entirety). It is widely used to measure muscle strength in eight proximal, distal, and axial muscle groups. Additionally, muscle strength testing, when assessed by the MMT-8, is also part of the response criteria for DM, PM (Rider et al., 2010), and JDM (Rider et al., 2018).

[0252] The MMT-8 is the method of choice for assessing muscle strength in more than 90% of IIM clinical trials because it is feasible, inexpensive, easy to perform, requires no equipment, and has adequate inter- and intra-rater reliability and validity when administered by trained examiners ( Rider et al., 2007 ; Saygin and Oddis, 2022 ).

[0253] IIM IIM is a diverse group of autoimmune diseases characterized by chronic muscle inflammation and associated weakness. IIM is a complex systemic disease with frequent manifestations in skeletal muscle and other organ systems, including the skin, joints, cardiopulmonary, gastrointestinal, and structural systems. As mentioned above, interactions between genetic and environmental factors are thought to lead to the development of distinct phenotypes of IIM.

[0254] The current paradigm explaining the pathophysiology of DM is that it results from an autoimmune attack on affected organs, which can be triggered by environmental factors such as UV exposure, drugs, infections, and lifestyle decisions in genetically predisposed individuals (Bogdanov et al., 2018). Initial pathogenic events are thought to occur in the endothelium of myoendothelial blood vessels (Bogdanov et al., 2018). When antibodies or other components activate C3, C3b and C4b fragments are formed, leading to the formation of C3bNEO and the membrane attack complex. These are then deposited in the myoendothelial vasculature, resulting in inflammation and infarction of the microvasculature, causing muscle atrophy. The pathophysiology of skin lesions is not fully understood, but the same mechanism has been proposed (Bogdanov et al., 2018). Several environmental factors, including UV exposure, drugs, infection, and lifestyle choices, may play a role in the pathophysiology of this disease. Associations with HLA DRB1*0301 and DQA1*0501 in Caucasians and with HLA-B7 in Asians have been reported, suggesting that there may be a genetic predisposition to DM. One leading hypothesis is that there is a pathogenic overproduction of IFN-β message and protein in the blood, muscle, and skin of DM patients, which appears to cause damage to these tissues.

[0255] Given that the group generally classified as PM encompasses patients who can be further subdivided based on MSA into ASyS, IMNM, and pure PM patients without specific autoantibodies, the pathophysiology may vary among these subgroups. The pathophysiology of all of these is not fully understood. In patients with PM and ASyS-associated myositis, an adaptive immune response driven by B cell responses has been observed, which has in turn been associated with the presence of anti-Jo1 autoantibodies in these patients. Subsequent findings suggest that anti-Jo-1 autoantibodies bind to a common autoepitope whose titer varies with disease activity, and that the immune response is associated with the clinical symptoms of myositis. Patients in this group have also been found to have autoantibodies recognizing HMGCR or SRP. HLA class II DRB1*11:01 is present in more than 70% of patients with IMNM, suggesting that it is a very strong risk factor for the development of autoimmune diseases. Muscle biopsies from these patients often contain areas of perifascicular necrosis (more common than in DM) and endomysial infiltration by T cells. Muscle tissue from patients who test positive for autoantibodies to HMGCD or SRP are histologically very similar. High titers of these autoantibodies have been observed to correlate with elevated CK, which is released into the bloodstream when muscle fibers are damaged (Lundberg et al., 2021).

[0256] Classification criteria have been developed and used to identify homogeneous and comparable patient populations. In 1975, Bohan and Peters laid the foundation by providing the first set of IIM classification criteria, which divided IIM into five groups. Since then, multiple classification and / or diagnostic criteria have been developed, but these have not been fully validated, and classification criteria continue to evolve. In 2017, validated classification criteria were approved and published by the ACR and EULAR (Lundberg et al., 2017). The ACR / EULAR classification criteria have been validated for adult and juvenile IIM. They define minimal, essential, and readily available clinical and laboratory characteristics to identify patients with IIM and distinguish them from those with non-IIM conditions with high sensitivity and specificity. In addition, they categorize IIM into major subgroups. These criteria were developed using a cohort of 976 IIM and 624 non-IIM cases from 47 centers worldwide. Variables were compiled from published criteria and expert consensus. The ACR / EULAR criteria classify patients as having "definite," "probable," and "suspected" disease based on a disease score and corresponding probability (≥50 but <55% = suspected, ≥55% but <90% = probable, ≥90% = definite). The IIM subclassification distinguishes between adult and juvenile myositis, polymyositis, dermatomyositis, DM without muscle symptoms, or inclusion body myositis.

[0257] overview PF-06823859 is a potent, selective humanized IgG1 neutralizing antibody directed against the human soluble cytokine IFNβ, a member of the type I IFN family of cytokines.

[0258] In a widely accepted model of type I IFN production (Noppert et al., 2007), stimulation of selective pattern recognition receptors (e.g., TLRs; DExD / H box RNA helicases such as RIG-I and MDA5; cGAS, which activates STING) leads to the dimerization and activation of the IFN regulatory factor IRF3, which then translocates into the nucleus and activates transcription at the IFNβ (and IFNα4) promoter. The IFNβ protein then signals in an autocrine and paracrine manner via binding to IFNAR. This induces intracellular signaling events downstream of IFNAR, leading to the expression of IRF7. IRF7 is then required for the transcription of multiple IFNα subtypes. Thus, in this model, IFNβ production initiates IFNAR signaling and concomitantly initiates IFNα production. Initial TLR activation can occur upon exposure to microbially derived PAMPs, including microbial nucleic acids, lipids, proteins, and lipoproteins. However, there is growing evidence that TLRs can also be stimulated by endogenous self-components (often referred to as damage-associated molecular patterns, or DAMPs) released during disease processes. It is noteworthy that IRF7 expression is constitutive in pDCs, and that TLR activation in pDCs directly leads to abundant expression of IFN isoforms.

[0259] This disclosure provides results from a Phase 2, multistage study to evaluate the efficacy and safety of PF-06823859 compared to placebo after 12 weeks of treatment in patients with dermatomyositis (DM) with either dermatomyositis-predominant (Stage 1, Stage 2, and corrected Stage 2) or myopathy-predominant (Stage 3) dermatomyositis. Among 18 participants with myopathy-predominant DM, the intended primary efficacy endpoint, mean Total Improvement Score (TIS), trended toward increasing over time (weeks 4 to 12) with no plateau until week 12, demonstrating numerical superiority of PF-06823859 600 mg compared to placebo. The estimated mean (90% confidence interval (CI)) TIS for PF-06823859 600 mg and placebo were 56.4 (41.4, 71.4) and 36.9 (22.0, 52.0), respectively, for a placebo-adjusted difference of 19.4 (-1.8, 40.7). Other endpoints related to muscle pathology, such as mean change from baseline (CFB) in manual muscle testing (MMT-8), mean CFB in the patient global assessment of myositis (PtGA), and mean CFB in the muscle enzyme creatine kinase (CK), also demonstrated numerical superiority for PF-06823859 600 mg compared with placebo at week 12.

[0260] Estimated mean CFB (90% CI) for muscle endpoints at week 12 were as follows: 1. MMT-8: PF-06823859 600 mg: 21.2 (11.9, 30.6); placebo: 11.7 (2.3, 21.0); placebo-adjusted difference: 9.6 (-3.8, 23.0). 2. PtGA (cm): PF-06823859 600 mg -4.6 (-5.9, -3.4) vs. placebo -1.2 (-2.4, 0.1); placebo-adjusted difference -3.5 (-5.3, -1.6). 3.CK (U / L): PF-06823859 600 mg -185.8 (-273.9, -97.6), placebo -39.9 (-125.7, 46.0), placebo-adjusted difference -145.9 (-269.4, -22.4).

[0261] For participants with dermatosis-predominant DM (including participants from stage 1 [n = 32], stage 2 [n = 9], and corrected stage 2 [n = 16]), the primary efficacy endpoint of interest was the Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A). The POC criteria for stage 1 were (i) a target value (TV) or treatment effect below -5, and (ii) the upper limit of the two-sided 90% CI of the treatment effect below 0.

[0262] The estimated CFB (90% CI) for the CDASI activity score (CDASI-A) at week 12 was as follows: 600 mg PF-06823859: -19.2 (-21.5, -16.8), placebo-adjusted difference -16.2 (-20.4, -12.1); 150 mg PF-06823859: -16.6 (-19.8, -13.4), placebo-adjusted difference -13.7 (-18.3, -9.0); Placebo: -2.9 (-6.3, 0.45).

[0263] Data from the skin cohort show that PF-06823859 600 mg and 150 mg were different from placebo (p<0.0001 for both doses), with similar treatment effects between doses. For patients with a CDASI-A score greater than 14 points, a 40% change in CDASI-A score indicates a meaningful change in quality of life (Ahmed et al., 2020), and PF-06823859 achieved this reduction in CDASI-A score in more than 80% of subjects.

[0264] In summary, both doses of PF-06823859 (600 mg and 150 mg) met (exceeded target) the primary efficacy endpoint for the skin cohort as assessed by CDASI-A score. Although the study was not powered for efficacy assessments for the muscle cohort, PF-06823859 600 mg had numerically better efficacy scores than placebo across all muscle function endpoints (TIS, MMT-8, PtGA for myositis, and CK).

[0265] safety There were two treatment discontinuations due to adverse events, both in the 600 mg group (one subject with microscopic colitis in stage 1 and one subject with leukopenia, cytopenia, and elevated liver enzymes in stage 3).

[0266] TEAEs were numerically higher and generally milder in severity in the active group compared with the placebo group across all stages. TEAEs observed in Stage 2 and corrected Stage 2 were not dose-related. The most common TEAEs by SOC in this study were infections and infestations, laboratory tests, and nervous system disorders. There were three treatment-related mild infections in the PF-06823859 group compared with two cases in the placebo group across all stages. There were four cases of positive SARS-CoV-2 testing in the placebo group and one suspected COVID-19 case in the PF-06823859 600 mg group. There were no cases of shingles or herpes simplex. Five participants reported seven SAEs: three in stage 1 and four in stage 3 (humerus fracture (600 mg group)); one subject in the 600 mg group reported three SAEs (leukopenia, cytopenia, and increased liver enzymes). There were no deaths or serious adverse reactions (SARs). Additionally, there were no clinically significant changes from baseline in safety laboratory findings, ECGs, and vital signs after treatment with PF-06823859.

[0267] In summary, PF-06823859 at 150 mg and 600 mg given monthly for a total of three doses was generally well tolerated and safe at all stages of the study.

[0268] Pharmacokinetics The PK of PF-06823859 IV was consistent across disease states and healthy participants. In patients with primarily dermatological disease, there was an approximately dose-proportional increase in exposure across the dose range of 150 mg Q4W to 600 mg Q4W. IP-10, a mechanistic biomarker of IFNb inhibition, decreased over time, with comparable mean percent reductions at week 12 in patients with cutaneous-predominant DM receiving either PF-06823859 150 mg or 600 mg. The exposure-response relationship appeared to saturate at 150 mg. The overall low incidence of drug-induced ADAs (3 / 22 at 600 mg in Stage 1) indicates a lack of clinically relevant impact on safety, PK, PD, or efficacy. No events of anaphylaxis or immunologically related clinical reactions of concern were observed. Clinical trial simulations confirmed that PF-06823859 achieved the pre-specified targeted product profile efficacy criteria at 24 weeks for TIS, MMT8, and CDASI-A.

[0269] In some embodiments, and without wishing to be bound by theory, it is postulated that PM patients may rely on the medication regimens of DM patients because proximal muscle weakness is a cardinal feature of both DM and PM, and the observation that type 1 IFN-inducible genes are elevated in the blood and muscle from patients with PM (albeit to a lesser extent than in DM).

[0270] Similarly, the similarities in clinical symptoms and IFN gene expression between JDM and adult DM populations suggest that adult dosing regimens will be safe, effective, and safe in adolescents. In some embodiments, the adolescent is at least 12 years old. In some embodiments, the adolescent weighs at least 30 kg. In some embodiments, the adolescent weighs at least 40 kg.

[0271] PK, PD, and efficacy modeling in DM participants supports the selection of 600 mg Q4W.

[0272] Inhibition of IP-10 was high at the 150 mg and 600 mg Q4W doses, with a small increase in % inhibition of IP-10 predicted between the 150 mg and 600 mg doses. IP-10 is a downstream biomarker of IFN-β inhibition that correlates with inflammation.

[0273] Regarding CDASI-A clinical efficacy, a lower dose of 150 mg Q4W demonstrated near-complete target binding in the skin-predominant cohort, but the same dose was not studied in the muscle-predominant cohort. Due to limited understanding of the correlation between the two cohorts, it is not possible to extrapolate muscle efficacy endpoints based on skin efficacy endpoints.

[0274] IFNβ binds highly (>99%) to PF-06823859 at exposures corresponding to the lower dose of 150 mg Q4W (assessed in cutaneous-predominant DM in Stage 2), demonstrating near-complete target binding at 150 mg Q4W and higher doses. This is consistent with the clinical efficacy response observed in terms of reduction in CDASI-A scores for cutaneous-predominant disease, which was similar between the 150 mg and 600 mg Q4W dose regimens.

[0275] In the Phase 2b study presented herein, the primary efficacy endpoint was the CDASI-A score. Estimated change from baseline in CDASI-A score at week 12 showed that 150 mg or 600 mg of PF-06823859 was different from placebo, with similar treatment effects between doses (p<0.0001 for both doses). Specifically, patients receiving 600 mg Q4W had a placebo-adjusted difference from baseline in CDASI-A score of -16.3, while patients receiving 150 mg Q4W had a placebo-adjusted difference in CDASI-A score of -13.7. In addition, PF-06823859 600 mg had numerically better efficacy scores than placebo across all key muscle function endpoints (TIS, MMT8, PtGA for myositis, and CK), with nominal statistical significance for PtGA (p=0.0046) and CK (p=0.0282). Both doses of PF-06823859 (150 mg and 600 mg) administered every 4 weeks in Study C0251002 were generally well tolerated and safe at all stages of the study. Additionally, no safety signals were identified, and no dose-related relationship was observed for all causalities of TEAEs at all stages in the study presented herein.

[0276] DM patient PK, PD, and efficacy modeling in DM participants from the Phase 2b study presented herein supports the selection of the 600 mg Q4W regimen for the Phase 3 study. Data obtained from Phase 1 and Phase 2 studies enabled the development of a PK / PD model population to characterize the relationship between drug exposure, target binding (total IFNβ, GS), and the PD biomarker IP-10 (see Examples). Given the near-complete understanding of target binding obtained through 600 mg Q4W PK / PD modeling, the high specificity of PF-06823859 for IFNβ, the acceptable safety profile, and the absence of off-target safety concerns observed from previous studies (primarily C0251002), the 600 mg Q4W regimen provides high confidence in efficacy against both skin and muscle endpoints.

[0277] Additionally, we developed exposure-response models to characterize the relationship between PF-06823859 exposure and clinical efficacy endpoints for skin and muscle. Modeling results support the following key conclusions for participants with DM: IFNβ binds highly (>99%) to PF-06823859 at exposures corresponding to the lower dose of 150 mg Q4W (assessed in cutaneous-predominant DM in Stage 2), demonstrating near-complete target binding at 150 mg Q4W and higher doses. This is consistent with the clinical efficacy response observed in terms of reduction in CDASI-A scores for cutaneous-predominant disease, which was similar between the 150 mg and 600 mg Q4W dose regimens. Inhibition of IP-10 was high at the 150 mg and 600 mg Q4W doses, with a small increase in % inhibition of IP-10 predicted between the 150 mg and 600 mg doses. IP-10 is a downstream biomarker of IFN-β inhibition that correlates with inflammation. Regarding CDASI-A clinical efficacy, a lower dose of 150 mg Q4W demonstrated near-complete target binding in the skin-predominant cohort, but the same dose was not studied in the muscle-predominant cohort. Due to limited understanding of the correlation between the two cohorts, it is not possible to extrapolate muscle efficacy endpoints based on skin efficacy endpoints.

[0278] PM patient PM and DM patients share a key characteristic: muscle weakness. Furthermore, type 1 IFN-inducible genes are elevated in the blood and muscle from patients with PM. Given the high level of target binding observed in DM patients and the demonstrated clinical efficacy of a 600 mg Q4W dose, it is predicted that the same dose will achieve a similar level of efficacy in PM patients. In any case, the sample size of the PM cohort is slightly larger than that of DM / JDM therapy, thereby allowing for a slightly lower efficacy response compared to that in DM / JDM and other indications.

[0279] SLE and other indications Based on the analysis provided herein, PF-06823859 shows surprisingly good potential for application in several additional indications, particularly SLE, cutaneous lupus, and psoriasis, and shows potential as a precision medicine for ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma.

[0280] In some embodiments, the disclosure provides a method for treating IIM in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of IIM by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0281] In some embodiments, the IIM is one or more conditions selected from the group consisting of dermatomyositis, polymyositis, inclusion body myositis, and juvenile dermatomyositis. In some embodiments, the IIM is dermatomyositis. In some embodiments, the IIM is polymyositis. In some embodiments, the IIM is inclusion body myositis. In some embodiments, the IIM is juvenile dermatomyositis.

[0282] In some embodiments, the disclosure provides a method for treating a patient having one or more conditions selected from the group consisting of SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen includes multiple individual doses that are at least two weeks apart from each other.

[0283] In some embodiments, the disclosure provides a method for treating a patient having one or more conditions selected from the group consisting of SLE, cutaneous lupus, and psoriasis, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen includes multiple individual doses that are at least two weeks apart from each other.

[0284] In some embodiments, the disclosure provides high-precision methods for treating a patient having one or more conditions selected from the group consisting of ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen includes multiple individual doses that are at least two weeks apart from each other.

[0285] In some embodiments, the disclosure provides a method for treating SLE in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to ameliorate signs and symptoms of SLE by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0286] In some embodiments, the disclosure provides a method for treating cutaneous lupus in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to improve signs and symptoms of cutaneous lupus by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses that are at least two weeks apart from each other.

[0287] In some embodiments, the disclosure provides a method for treating psoriasis in a patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to improve signs and symptoms of psoriasis by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least two weeks.

[0288] In some embodiments, the individual doses are separated from each other by a period selected from the group consisting of at least 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, and 3 months. Preferably, the individual doses are separated from each other by the same time interval.

[0289] In some embodiments, one or more of the individual doses is an amount within a range having a lower limit selected from the group consisting of about 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, and 600 mg, and an upper limit selected from the group consisting of 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg. The dose may be, or may be at least, an amount selected from the group consisting of 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg. In some embodiments, the dose is 150 mg. In some embodiments, the dose is 300 mg. In some embodiments, the dose is about 600 mg. In some embodiments, the dose is about 900 mg. Preferably, each dose is the same amount.

[0290] In some embodiments, the dosing regimen continues for a period of time selected from the group consisting of about 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, 3 months, 16 weeks, 4 months, 20 weeks, 5 months, 24 weeks, 6 months, and 26 weeks, or at least such a period.

[0291] In some embodiments of the present disclosure, improvement in signs or symptoms is assessed at a time point from the start of the dosing regimen selected from the group consisting of about 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, 3 months, 16 weeks, 4 months, 20 weeks, 5 months, 24 weeks, 6 months, and 26 weeks.

[0292] In some embodiments of the present disclosure, the improvement of signs or symptoms is maintained for a maintenance period, which begins at a time from the start of the dosing regimen selected from the group consisting of about 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, 3 months, 16 weeks, 4 months, 20 weeks, 5 months, 24 weeks, 6 months, and 26 weeks. In some embodiments of the present disclosure, the maintenance period is selected from the group consisting of 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, 3 months, 16 weeks, 4 months, 20 weeks, 5 months, 24 weeks, 6 months, and 26 weeks.

[0293] In some embodiments of the present disclosure, at least 4 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms, which is maintained for an additional period of time selected from the group consisting of at least about 4 weeks, 6 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks.

[0294] In some embodiments of the present disclosure, at least 8 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms, which is maintained for an additional period of time selected from the group consisting of at least about 4 weeks, 6 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks.

[0295] In some embodiments of the present disclosure, at least 12 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms, which is maintained for an additional period of time selected from the group consisting of at least about 4 weeks, 6 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks.

[0296] In some embodiments of the present disclosure, at least 16 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms, which is maintained for an additional period of time selected from the group consisting of at least about 4 weeks, 6 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks.

[0297] In some embodiments of the present disclosure, at least 24 weeks after initiation of the dosing regimen, the patient experiences improvement in signs and symptoms, which is maintained for an additional period of time selected from the group consisting of at least about 4 weeks, 6 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks.

[0298] In some embodiments of the present disclosure, improvement in signs or symptoms is characterized by a clinical response.

[0299] Clinical response was (i) change from baseline in manual muscle testing (MMT-8) score greater than 0; (ii) a total improvement score (TIS) greater than 0; (iii) change from baseline in patient global assessment score below 0; (iv) improvement of absolute values ​​of the muscle enzyme creatinine kinase below zero; and (v) Change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) below 0 The composition may be characterized by one or more means selected from the group consisting of:

[0300] In some embodiments of the present disclosure, the change from baseline in manual muscle testing (MMT-8) is characterized by an MMT-8 score of at least a value selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0301] In some embodiments of the present disclosure, improvement in Total Improvement Score (TIS) is characterized by a TIS value selected from the group consisting of at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, and 70. A clinical response defined as minimal improvement in DM includes a TIS of at least 20. A clinical response defined as minimal improvement in JDM includes a TIS of at least 30. A clinical response defined as moderate improvement in DM includes a TIS of at least 40. A clinical response defined as moderate improvement in JDM includes a TIS of at least 45. A clinical response defined as major improvement in DM includes a TIS of at least 60. A clinical response defined as major improvement in JDM includes a TIS of at least 70. The TIS may be placebo-corrected. When a clinical response is characterized by a placebo-corrected TIS, the TIS is greater than 0.

[0302] In some embodiments of the present disclosure, the change from baseline in Patient Global Assessment is characterized by a PtGA score not exceeding a value selected from the group consisting of -1, -2, -3, -4, and -5.

[0303] In some embodiments of the present disclosure, the change from baseline in the absolute muscle enzyme creatine kinase is characterized by an amount not greater than a value selected from the group consisting of −75, −80, −85, −90, −95, −100, −105, −110, −115, −120, −125, −130, −135, −140, −145, −150, −155, −160, −165, −170, −175, −180, and −185.

[0304] In some embodiments of the present disclosure, the change from baseline in the Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) is characterized by a CDASI-A score not exceeding a value selected from the group consisting of -6, -7, -8, -9, -10, -11, -12, -13, -14, -15, -16, -17, -18, -19, and -20.

[0305] In some embodiments of the present disclosure, the patient experiences improvement in signs or symptoms within a time period selected from the group consisting of about 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks after starting the dosing regimen.

[0306] In some embodiments of the present disclosure, the patient demonstrates a clinical response within a time period selected from the group consisting of about 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks after starting the dosing regimen.

[0307] In some embodiments of the present disclosure, the anti-IFNβ antibody comprises three CDRs derived from a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and three CDRs derived from a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0308] In some embodiments of the present disclosure, the anti-IFNβ antibody comprises an HCDR1 having the sequence set forth in SEQ ID NO: 5, an HCDR2 having the sequence set forth in SEQ ID NO: 6, an HCDR3 having the sequence set forth in SEQ ID NO: 7, an LCDR1 having the sequence set forth in SEQ ID NO: 8, an LCDR2 having the sequence set forth in SEQ ID NO: 9, and an LCDR3 having the sequence set forth in SEQ ID NO: 10.

[0309] In some embodiments of the present disclosure, the anti-IFNβ antibody comprises a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0310] In some embodiments of the disclosure, the anti-IFNβ antibody comprises a heavy chain having the sequence set forth in SEQ ID NO: 1 and a light chain having the sequence set forth in SEQ ID NO: 2. In some embodiments of the disclosure, the C-terminal lysine (K) of the heavy chain amino acid sequence of SEQ ID NO: 1 is optional.

[0311] In some embodiments of the present disclosure, the anti-IFNβ antibody comprises a VH encoded by the nucleic acid sequence of an insert in the vector deposited with ATCC as CTI-AF1-VH with ATCC accession number PTA-122727, and a VL encoded by the nucleic acid sequence of an insert in the vector deposited with ATCC as CTI-AF1-VL with ATCC accession number PTA-122726. In some embodiments of the present disclosure, the anti-IFNβ antibody comprises a VH sequence encoded by an insert in a plasmid deposited with ATCC with ATCC accession number PTA-122727. In some embodiments of the present disclosure, the anti-IFNβ antibody comprises a VL sequence encoded by an insert in a plasmid deposited with ATCC with ATCC accession number PTA-122726.

[0312] In some embodiments of the present disclosure, the anti-IFNβ antibody competes for binding with an anti-IFNβ antibody comprising a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4.

[0313] In some embodiments of the present disclosure, the anti-IFNβ antibody competes for binding with an antibody comprising a VH encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VH with ATCC accession number PTA-122727 and a VL encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VL with ATCC accession number PTA-122726.

[0314] In some embodiments of the present disclosure, the anti-IFNβ antibodies can be used in the preparation of a medicament for a method of treatment with any of those provided herein.

[0315] Common techniques The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill of a person of ordinary skill in the art. Such techniques are described in the literature, e.g., Molecular Cloning: A Laboratory Manual, 2nd Edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (eds. JM Miller and MPCalos, 1987); Current Protocols in Molecular Biology (eds. FMAusubel et al., 1987); PCR: The Polymerase Chain Reaction (eds. Mullis et al., 1994); Current Protocols in Immunology (eds. JE Coligan et al., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P.Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D. Catty, ed., IRL Press, 1988-1989); Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and JD Capra, eds., Harwood Academic Publishers, 1995).

[0316] definition The following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0317] An "antibody" is an immunoglobulin molecule capable of specifically binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site located in the variable region of the immunoglobulin molecule. As used herein, the term encompasses not only intact polyclonal or monoclonal antibodies, but also, unless otherwise indicated, any antigen-binding portion thereof that competes with the intact antibody for specific binding, fusion proteins containing an antigen-binding portion, and any other modified configuration of an immunoglobulin molecule containing an antigen recognition site. Antigen-binding portions include, for example, Fab, Fab', F(ab')2, Fd, Fv, domain antibodies (dAbs, e.g., shark and camel antibodies), fragments containing complementarity-determining regions (CDRs), single-chain variable fragment antibodies (scFv), maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs, as well as polypeptides comprising at least a portion of an immunoglobulin sufficient to confer specific antigen binding to the polypeptide. Antibodies include antibodies of any class, e.g., IgG, IgA, or IgM (or subclasses thereof); antibodies need not be of any particular class. Immunoglobulins can be assigned to different classes depending on the antibody amino acid sequence of the constant region of their heavy chains. There are five major immunoglobulin classes: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0318] The "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. As is known in the art, the variable regions of the heavy and light chains each consist of four framework regions (FRs) connected by three complementarity-determining regions (CDRs), also known as hypervariable regions, which contribute to the formation of the antigen-binding site of the antibody. When a variant of the target variable region is desired, particularly one that involves the substitution of amino acid residues outside the CDR region (i.e., within the framework region), appropriate amino acid substitutions, preferably conservative amino acid substitutions, can be identified by comparing the target variable region with the variable regions of other antibodies that contain CDR1 and CDR2 sequences within the same canonical class as the target variable region (Chothia and Lesk, J Mol Biol 196(4):901-917, 1987).

[0319] In certain embodiments, definitively defining CDRs and identifying the residues comprising the antibody binding site are achieved by analyzing the structure of the antibody and / or the structure of the antibody-ligand complex. In certain embodiments, this can be achieved by any of a variety of techniques known to those skilled in the art, such as X-ray crystallography. In certain embodiments, various analytical methods can be used to identify or estimate CDR regions. Examples of such methods include, but are not limited to, the Kabat definition, the Chothia definition, the AbM definition, the contact definition, and the conformational definition.

[0320] The Kabat definition is a standard method for numbering residues in antibodies and is typically used to identify CDR regions. See, e.g., Johnson and Wu, 2000, Nucleic Acids Res., 28:214-8. The Chothia definition is similar to the Kabat definition, but takes into account the location of certain structural loop regions. See, e.g., Chothia et al., 1986, J. Mol. Biol., 196:901-17; Chothia et al., 1989, Nature, 342:877-83. The AbM definition uses an integrated suite of computer programs developed by the Oxford Molecular Group to model antibody structure. See, e.g., Martin et al., 1989, Proc Natl Acad Sci (USA), 86:9268-9272; "AbM™, A Computer Program for Modeling Variable Regions of Antibodies," Oxford, UK; Oxford Molecular, Ltd. The AbM definition models the tertiary structure of antibodies from primary sequence using a combination of knowledge databases and first-principles methods, e.g., those described by Samudrala et al., 1999, "Ab Initio Protein Structure Prediction Using a Combined Hierarchical Approach," in PROTEINS, Structure, Function and Genetics Suppl., 3:194-198. The contact definition is based on an analysis of available complex crystal structures. See, e.g., MacCallum et al., 1996, J. Mol. Biol., 5:732-45. In another approach, referred to herein as "conformational definition" of CDRs, CDR positions can be identified as residues that make enthalpic contributions to antigen binding. See, e.g., Makabe et al., 2008, Journal of Biological Chemistry, 283:1156-1166.Still other CDR boundary definitions may not strictly follow any of the above approaches, but will still overlap at least a portion of the Kabat CDRs, but they may be shortened or extended based on predictions or experimental findings that certain residues or groups of residues do not significantly affect antigen binding. As used herein, CDRs may refer to CDRs defined by any approach known in the art, including a combination of approaches. The methods used herein may utilize CDRs defined according to any of these approaches. For any given embodiment that includes more than one CDR, the CDRs may be defined according to any of the Kabat, Chothia, extended, AbM, contact, and / or conformational definitions.

[0321] As known in the art, a "constant region" of an antibody refers to the constant region of the antibody light chain or the constant region of the antibody heavy chain, either alone or in combination.

[0322] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population; i.e., the individual antibodies comprising the population are identical except for possible minor naturally occurring mutations. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous antibody population and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be produced by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or may be produced by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. The monoclonal antibodies may also be isolated from phage libraries generated using the techniques described in McCafferty et al., 1990, Nature 348:552-554, for example.

[0323] As known in the art, the terms "polynucleotide" and "nucleic acid," used interchangeably herein, refer to a nucleotide chain of any length, including DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a chain by DNA or RNA polymerase. Polynucleotides can contain modified nucleotides, such as methylated nucleotides and their analogs. If present, modifications to the nucleotide structure can be imparted before or after assembly of the chain. The sequence of nucleotides can be interrupted by non-nucleotide components. Polynucleotides can be further modified after polymerization, for example, by conjugation with a labeling component. Other types of modifications include, for example, "caps," substitutions of one or more of the naturally occurring nucleotides with analogs, internucleotide modifications, such as, for example, those with uncharged linkages (e.g., methyl phosphate, phosphotriester, phosphoamidate, carbamate, etc.) and charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant moieties, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those with intercalators (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, metal oxides, etc.), those containing alkylators, those with modified linkages (e.g., alpha-anomeric nucleic acids, etc.), and unmodified forms of polynucleotides. Additionally, any of the hydroxyl groups normally present on the sugar may be replaced by, for example, phosphonate groups, phosphate groups, protected by standard protecting groups, or activated to prepare additional linkages to additional nucleotides, or conjugated to a solid support. The 5' and 3' terminal OH can be phosphorylated or substituted with amines or organic capping group moieties of 1 to 20 carbon atoms. Other hydroxyls can also be derivatized to standard protecting groups.Polynucleotides can also contain analog forms of ribose or deoxyribose sugars commonly known in the art, including, for example, 2'-O-methyl-, 2'-O-allyl, 2'-fluoro-, or 2'-azido-ribose, carbocyclic sugar analogs, alpha- or beta-anomeric sugars, epimeric sugars such as arabinose, xylose, or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acyclic analogs, and abasic nucleoside analogs, such as methyl riboside. One or more phosphodiester linkages may be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, embodiments in which the phosphate is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), (O)NR2 ("amidate"), P(O)R, P(O)OR', CO, or CH2 ("formacetal"), where each R or R' is independently H, or substituted or unsubstituted alkyl (1-20 C), aryl, alkenyl, cycloalkyl, cycloalkenyl, or araldyl, which may contain an ether (-O-) linkage. Not all linkages in a polynucleotide need be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0324] An antibody that "preferentially binds" or "specifically binds" (used interchangeably herein) to an epitope is a term well understood in the art, and methods for determining such specific or preferential binding are also well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular cell or substance more frequently, more rapidly, for a longer period, and / or with a higher affinity than with another cell or substance. An antibody "specifically binds" or "preferentially binds" to a target if it binds with higher affinity, avidity, more readily, and / or for a longer period than it binds to other substances. For example, an antibody that specifically or preferentially binds to a target (e.g., INFβ) epitope is an antibody that binds to this epitope with higher affinity, avidity, more readily, and / or for a longer period than it binds to other target epitopes or non-target epitopes. It is understood by reading this definition that, for example, an antibody (or moiety or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Generally, reference to binding implies preferential binding, but this is not necessarily the case.

[0325] As used herein, "substantially pure" refers to a material that is at least 50% pure (i.e., free from contaminants), more preferably at least 90% pure, more preferably at least 95% pure, even more preferably at least 98% pure, and most preferably at least 99% pure.

[0326] A "host cell" includes an individual cell or cell culture that can be or has been a recipient of a vector for incorporation of a polynucleotide insert. A host cell includes the progeny of a single host cell, and the progeny may not necessarily be completely identical (in terms of morphology or genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. A host cell includes cells transfected in vivo with a polynucleotide of the invention.

[0327] As known in the art, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain. The "Fc region" may be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is usually defined as stretching from the amino acid residue at position Cys226 or Pro230 to the carboxyl terminus. The numbering of residues within the Fc region is that of the EU index in Kabat. Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991. The Fc region of an immunoglobulin generally contains two constant domains, CH2 and CH3. As known in the art, the Fc region can exist in a dimeric or monomeric form.

[0328] As used in the art, "Fc receptor" and "FcR" refer to a receptor that binds to the Fc region of an antibody. A preferred FcR is a native-sequence human FcR. Additionally, a preferred FcR is one that binds IgG antibodies (gamma receptors), including receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA (an "activating receptor") and FcγRIIB (an "inhibitory receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. FcRs are discussed in Ravetch and Kinet, 1991, Ann. Rev. Immunol., 9:457-92; Capel et al., 1994, Immunomethods, 4:25-34; and de Haas et al., 1995, J. Lab. Clin. Med., 126:330-41. "FcR" also includes the fetal receptor, FcRn, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., 1976, J. Immunol., 117:587 and Kim et al., 1994, J. Immunol., 24:249).

[0329] The term "compete," as used herein with respect to antibodies, means that a first antibody, or antigen-binding portion thereof, binds to an epitope in a manner sufficiently similar to that of a second antibody, or antigen-binding portion thereof, such that the result of binding of the first antibody to its cognate epitope in the presence of the second antibody is detectably reduced compared to binding of the first antibody in the absence of the second antibody. Alternatively, binding of the second antibody to its epitope may, but need not, also be detectably reduced in the presence of the first antibody. That is, a first antibody may inhibit binding of a second antibody to its epitope without the second antibody inhibiting binding of the first antibody to its respective epitope. However, if each antibody detectably inhibits binding of the other antibody to its cognate epitope or ligand, whether to the same, greater, or lesser extent, the antibodies are said to "cross-compete" with each other for binding of their respective epitopes. Both competing and cross-competing antibodies are encompassed by the present invention. Regardless of the mechanism by which such competition or cross-competition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope or portion thereof), one of skill in the art will understand, based on the teachings provided herein, that such competing and / or cross-competing antibodies are encompassed and may be useful in the methods disclosed herein.

[0330] As used herein, "treatment" refers to an approach for obtaining beneficial or desired clinical results. For purposes of the present invention, beneficial or desired clinical results include, for example, a reduction or amelioration of the signs and symptoms of osteoarthritis compared to before administration of an anti-IFNβ antibody.

[0331] "Alleviating" refers to, for example, a reduction or improvement in one or more signs or symptoms of osteoarthritis compared to not administering an anti-IFNβ antibody described herein. "Alleviating" also includes a shortening or reduction in the duration of symptoms.

[0332] As used herein, an "effective dosage" or "effective amount" of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve any one or more beneficial or desired results. In more specific embodiments, the effective amount prevents, reduces, or alleviates signs or symptoms of myositis and / or prolongs survival in the subject being treated. For prophylactic use, beneficial or desired results include eliminating or reducing the risk of, reducing the severity of, or delaying the onset of, the disease, its complications, and the biological, histological, and / or behavioral symptoms of intermediate pathological phenotypes that appear during the development of the disease. For therapeutic use, beneficial or desired results include clinical results, such as reducing one or more signs or symptoms of myositis, reducing the dose of other medications required to treat the disease, enhancing the effect of another medication, and / or delaying the progression of the disease in the patient. An effective dosage can be administered in one or more administrations. For the purposes of the present invention, an effective dosage of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve prophylactic or therapeutic treatment, either directly or indirectly.As understood in a clinical context, an effective dosage of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition.Therefore, an "effective dosage" can be considered in the context of administering one or more therapeutic agents, and a single agent can be considered to be provided in an effective amount if it can or will achieve a desired result in combination with one or more agents.

[0333] When the assessment of the signs or symptoms of IIM is quantified by clinical scales compared with baseline and during and / or after treatment, the treatment "effectively improves" or "effectively reduces." The difference between the clinical scales at baseline and those during / after treatment is compared and used to determine whether the signs or symptoms have improved and whether the treatment is effective. This comparison can include comparison with placebo or one or more of previous treatments.

[0334] A "patient," "individual," or "subject," as used interchangeably herein, is a mammal, more preferably a human. Mammals also include, but are not limited to, farm animals (e.g., cows, pigs, horses, chickens, etc.), sport animals, pets, primates, horses, dogs, cats, mice, and rats.

[0335] As used herein, " pharmaceutically acceptable carrier " or " pharmaceutically acceptable excipient " includes any material that, when combined with active ingredient, allows the active ingredient to maintain its biological activity and does not react with the subject's immune system. Examples include, but are not limited to, standard pharmaceutical carriers, such as phosphate buffered saline solution, water, emulsions, such as oil / water emulsions, and any of various types of wetting agents. The preferred diluent for aerosol or parenteral administration is phosphate buffered saline (PBS) or normal saline (0.9%). The composition containing such carriers is formulated by well-known conventional methods (see, for example, Remington's Pharmaceutical Sciences, 18th Edition, A. Gennaro, ed., Mack Publishing Co., Easton, PA, 1990 and Remington, The Science and Practice of Pharmacy, 20th Edition, Mack Publishing, 2000).

[0336] As used herein, reference to "about" a value or parameter includes (and describes) embodiments directed to that value or parameter itself. For example, a statement referring to "about X" includes the statement "X." Numerical ranges include the numbers defining the range. Generally speaking, the term "about" refers to the stated value of a variable, and all values ​​of that variable that fall within the stated value's experimental error (e.g., within a 95% confidence interval of the mean) or within 10 percent of the stated value, whichever is greater. When the term "about" is used in the context of a time period (years, months, weeks, days, etc.), the term "about" means the next subunit of that time period plus or minus one (e.g., about 1 year means 11-13 months; about 6 months means 6 months ± 1 week; about 1 week means 6-8 days, etc.), or within 10 percent of the stated value, whichever is greater.

[0337] The term "subcutaneous administration" refers to administration of a substance into the subcutaneous layer.

[0338] The term "preventing" or "preventing" refers to (a) keeping a disorder from occurring, or (b) delaying the onset of a disorder or the onset of symptoms of a disorder.

[0339] Where embodiments are described herein using the word "comprising," it is understood that similar embodiments separately described using the terms "consisting of" and / or "consisting essentially of" are also provided.

[0340] When aspects or embodiments of the invention are described in terms of a Markush group or other grouping of alternatives, the invention includes not only the entire group listed as a whole, but also each member of the group individually and all possible subgroups of the main group, as well as the main group in which one or more of the group members are absent. The invention also envisions the explicit exclusion of any one or more of the group members in the claimed invention.

[0341] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control. Throughout this specification and claims, the term "comprise" or variations such as "comprises" or "comprising" will be understood to imply the inclusion of a stated integer or group of integers, but not the exclusion of any other integer or group of integers. Unless otherwise required by context, singular terms shall include the plural, and plural terms shall include the singular. Any examples following the terms "e.g.," or "for example," are not meant to be exclusive or limiting.

[0342] Although exemplary methods and materials are described herein, methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0343] Anti-IFNβ antibody

[0344] [Table 3-1]

[0345] [Table 3-2]

[0346] The antibodies described herein can be produced by any method known in the art.For the production of hybridoma cell lines, the route and schedule of immunization of host animals generally follow established conventional techniques for antibody stimulation and production, as further described herein.General techniques for the production of human and mouse antibodies are known in the art and / or described herein.

[0347] It is contemplated that any mammalian subject, including a human, or antibody-producing cells derived therefrom can be engineered to serve as the basis for the production of mammalian, including human, and hybridoma cell lines. Typically, the host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and / or intradermally with an amount of an immunogen, including those described herein.

[0348] Hybridomas can be prepared from lymphocytes and immortalized myeloma cells using the general somatic cell hybridization technique of Kohler, B. and Milstein, C., Nature 256:495-497, 1975, or a modified version by Buck, DW et al., In Vitro, 18:377-381, 1982. Available myeloma lines, including but not limited to X63-Ag8.653 and those obtained from the Salk Institute, Cell Distribution Center, San Diego, Calif., USA, may be used for hybridization. Generally, this technique involves fusing myeloma and lymphoid cells using a fusogen, such as polyethylene glycol, or by electrical means well known to those skilled in the art. After fusion, the cells are separated from the fusion medium and grown in a selective growth medium, such as hypoxanthine-aminopterin-thymidine (HAT) medium, to eliminate unhybridized parent cells. Any of the media described herein, with or without serum supplementation, can be used to culture hybridomas secreting monoclonal antibodies. As another alternative to cell fusion techniques, EBV-immortalized B cells can be used to produce the monoclonal antibodies of the present invention. The hybridomas are expanded and, if desired, subcloned, and the supernatants are assayed for anti-immunogen activity by conventional immunoassay procedures (e.g., radioimmunoassay, enzyme immunoassay, or fluorescence immunoassay).

[0349] Hybridomas that can be used as a source of antibodies include all derivatives and progeny of the parent hybridoma that produce monoclonal antibodies.

[0350] Hybridomas producing antibodies used in the present invention may be grown in vitro or in vivo using known procedures. Monoclonal antibodies may be isolated from culture media or body fluids, if desired, by conventional immunoglobulin purification procedures, such as ammonium sulfate precipitation, gel electrophoresis, dialysis, chromatography, and ultrafiltration. Undesirable activity, if present, can be removed, for example, by passing the preparation through an adsorbent made from a solid-phase-bound immunogen, and eluting or releasing the desired antibody from the immunogen. Bifunctional or derivatizing agents, such as maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine ​​residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl2, or R 1 N=C=NR, where R and R 1 are different alkyl groups), antibody (e.g., monoclonal antibody) populations can be obtained by immunization of host animals with cells expressing the antibody target (e.g., IFNβ), a human target protein (e.g., IFNβ), or a fragment containing the target amino acid sequence, conjugated to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor.

[0351] If desired, the antibody of interest (monoclonal or polyclonal) can be sequenced, and the polynucleotide sequence can then be cloned into a vector for expression or expansion.The sequence encoding the antibody of interest can be maintained in a vector in host cells, and the host cells can then be expanded and frozen for future use.The production of recombinant monoclonal antibodies in cell culture can be carried out by cloning antibody genes from B cells by means known in the art.For example, see Tiller et al., J.Immunol.Methods 329, 112, 2008, U.S. Patent No. 7,314,622.

[0352] In some embodiments, antibodies can be produced using hybridoma technology. It is contemplated that any mammalian subject, including humans, or antibody-producing cells derived therefrom can be engineered to serve as the basis for the production of hybridoma cell lines, including mammals, including humans. The route and schedule for immunization of host animals generally follows established conventional techniques for antibody stimulation and production, as further described herein. Typically, host animals are inoculated intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and / or intradermally with an amount of immunogen, including those described herein.

[0353] In some embodiments, the antibodies described herein are glycosylated at conserved positions within their constant regions (Jefferis and Lund, 1997, Chem. Immunol. 65:111-128; Wright and Morrison, 1997, TibTECH 15:26-32). The oligosaccharide side chains of immunoglobulins affect protein function (Boyd et al., 1996, Mol. Immunol. 32:1311-1318; Wittwe and Howard, 1990, Biochem. 29:4175-4180), as well as intramolecular interactions between portions of the glycoprotein, which can affect the glycoprotein's organization and presented three-dimensional surface (Jefferis and Lund, supra; Wyss and Wagner, 1996, Current Opin. Biotech. 7:409-416). Oligosaccharides can also function to target a given glycoprotein to a specific molecule based on specific recognition structures. Glycosylation of antibodies has also been reported to affect antibody-dependent cellular cytotoxicity (ADCC). Specifically, antibodies produced by CHO cells with tetracycline-regulated expression of β(1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase that catalyzes the formation of bisecting GlcNAc, have been reported to have improved ADCC activity (Umana et al., 1999, Nature Biotech. 17:176-180).

[0354] Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine, asparagine-X-threonine, and asparagine-X-cysteine ​​are recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain, where X is any amino acid except proline. Thus, the presence of any of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of the sugar N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used.

[0355] Addition of glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by adding, or substituting, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).

[0356] The glycosylation pattern of an antibody can also be altered without changing the underlying nucleotide sequence. Glycosylation depends in large part on the host cell used to express the antibody. Because the cell types used to express recombinant glycoproteins, e.g., antibodies, as potential therapeutics, are rarely native cells, variations in the glycosylation pattern of antibodies are predictable (see, e.g., Hse et al., 1997, J. Biol. Chem. 272:9062-9070).

[0357] In addition to the choice of host cell, factors that affect glycosylation during recombinant production of antibodies include growth mode, media formulation, culture density, oxygenation, pH, purification scheme, etc. Various methods have been proposed to alter the glycosylation pattern achieved in a particular host organism, including introducing or overexpressing certain enzymes involved in oligosaccharide production (U.S. Pat. Nos. 5,047,335, 5,510,261, and 5,278,299). Glycosylation, or certain types of glycosylation, can be enzymatically removed from glycoproteins using, for example, endoglycosidase H (Endo H), N-glycosidase F, endoglycosidase F1, endoglycosidase F2, or endoglycosidase F3. In addition, recombinant host cells can be genetically engineered to be defective in processing certain types of polysaccharides. These and similar techniques are well known in the art.

[0358] Other modification methods include using linkage techniques known in the art, including, but not limited to, enzymatic means, oxidative substitution, and chelation. Modifications can be used, for example, for attaching labels for immunoassays. Modified polypeptides can be made using procedures established in the art and screened using standard assays known in the art, some of which are described below and in the Examples.

[0359] Polynucleotides, Vectors, and Host Cells The present invention also provides polynucleotides encoding any of the anti-IFNβ antibodies described herein. Polynucleotides can be made and expressed by means known in the art.

[0360] In another aspect, the invention provides compositions (e.g., pharmaceutical compositions) comprising any of the polynucleotides of the invention for use in one or more methods of the invention. In some embodiments, the composition comprises an expression vector comprising a polynucleotide encoding any of the anti-IFNβ antibodies described herein for use in one or more methods of the invention.

[0361] In another embodiment, an isolated cell line producing an anti-IFNβ antibody described herein for use in one or more methods of the present invention is provided.

[0362] Polynucleotides complementary to any such sequences are also encompassed by the present invention. Polynucleotides may be single-stranded (coding or antisense) or double-stranded, and may be DNA (genomic, cDNA, or synthetic) or RNA molecules. RNA molecules include HnRNA molecules, which contain introns and correspond in a one-to-one manner to DNA molecules, and intron-free mRNA molecules. Additional coding or non-coding sequences may, but need not, be present within a polynucleotide of the present invention, and polynucleotides may, but need not, be linked to other molecules and / or support materials.

[0363] The polynucleotide may comprise a native sequence (i.e., an endogenous sequence encoding an antibody or fragment thereof) or may comprise a variant of such a sequence. Polynucleotide variants contain one or more substitutions, additions, deletions, and / or insertions such that the immunoreactivity of the encoded polypeptide is not diminished compared to the native immunoreactive molecule. The effect on the immunoreactivity of the encoded polypeptide can generally be assessed as described herein. Variants preferably exhibit at least about 70% identity, more preferably at least about 80% identity, even more preferably at least about 90% identity, and most preferably at least about 95% identity to the polynucleotide sequence encoding a native antibody or fragment thereof.

[0364] composition The present invention also provides pharmaceutical compositions comprising an effective amount of the anti-IFNβ antibodies described herein, and such pharmaceutical compositions for use in the methods of treatment described herein.Examples of such compositions and methods of formulation are also described herein.It is understood that a composition can contain more than one anti-IFNβ antibody.

[0365] The compositions used in the present invention may further comprise a pharmaceutically acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or aqueous solution (Remington: The Science and Practice of Pharmacy, 20th Edition, 2000, Lippincott Williams and Wilkins, eds. K.E. Hoover). Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosage and concentration, and include buffering agents such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum The excipient may include albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextran; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG). Pharmaceutically acceptable excipients are further described herein.

[0366] The anti-IFNβ antibodies and compositions thereof can also be used in conjunction with, or administered separately, simultaneously, or sequentially with, other agents that function to enhance and / or support the effectiveness of the agent.

[0367] formulation The antibodies or antigen-binding fragments thereof of the present invention can be formulated as pharmaceutical compositions, which may further comprise pharmaceutically acceptable carriers, excipients, and / or stabilizers in the form of lyophilized formulations or aqueous solutions (Remington: The Science and Practice of Pharmacy, 20th Edition, 2000, Lippincott Williams and Wilkins, eds. K.E. Hoover). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations and include buffers, e.g., phosphate, citrate, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens, e.g., methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (fewer than about 10 residues) polypeptides; proteins, e.g., serum The excipient may include albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextran; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG). Pharmaceutically acceptable excipients are further described herein.

[0368] Therapeutic formulations of anti-IFNβ antibodies used in accordance with the present invention are prepared for storage by mixing the protein having the desired purity, in the form of a lyophilized formulation or aqueous solution, with optional pharmaceutically acceptable carriers, excipients, or stabilizers (Remington, The Science and Practice of Pharmacy 20th ed. Mack Publishing, 2000). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include, but are not limited to, buffering agents, e.g., phosphates, citrates, and other organic acids; salts, e.g., sodium chloride; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens, e.g., methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (fewer than about 10 residues) polypeptides; proteins The carrier may include proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0369] Preparations to be used for in vivo administration must be sterile. This is readily accomplished, for example, by filtration through sterile filtration membranes. Therapeutic anti-IFNβ antibody compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0370] In one embodiment, a stable aqueous formulation is provided comprising at least 25 mg / ml to about 200 mg / ml of an anti-IFNβ antibody or antigen-binding fragment thereof, a buffer, a polyol, a surfactant, a stabilizer, and optionally a chelating agent, the aqueous formulation having a pH of about 5.0 to about 6.5. The formulations described herein preferably have a long shelf life (e.g., at about 5°C), of at least about 36 months or more. The formulations described herein are particularly useful for use in the methods and uses described herein.

[0371] In some embodiments, the present disclosure provides an aqueous formulation containing an anti-IFNβ antibody at a concentration of about 25 mg / mL to about 200 mg / mL, histidine or His-HCl at a concentration of 10 to 50 mM, arginine or NaCl in an amount of 20 to 150 mM, and a polyol (preferably sucrose or trehalose in an amount of 20 mg / mL to 85 mg / mL), at a pH of 5.0 to 6.5. In some embodiments, the antibody may further optionally contain a chelating agent.

[0372] In some embodiments, the antibody may be selected from the group consisting of monoclonal antibodies, polyclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2, Fv, Fc, ScFv, etc.), chimeric antibodies, bispecific antibodies, heteroconjugate antibodies, single-chain (ScFv), mutants thereof, fusion proteins comprising antibody portions (e.g., domain antibodies), humanized antibodies, human antibodies, and any other modified configuration of an immunoglobulin molecule comprising an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. The antibody may be murine, rat, human, or of any other origin (including chimeric or humanized antibodies). In some embodiments, the antibody may be human, but is more preferably humanized. Preferably, the antibody is isolated, and even more preferably substantially pure. If the antibody is an antibody fragment, it preferably retains the functional characteristics of the original antibody, i.e., ligand binding and / or antagonist or agonist activity. In some embodiments, the antibody heavy chain constant region can be derived from any type of constant region, such as IgG, IgM, IgA, and IgE, and any isotype, such as IgG1, IgG2, IgG3, and IgG4. Preferably, the antibody is an IgG1 antibody.

[0373] In some embodiments, the IFNβ antibody comprises three CDRs derived from the variable heavy chain region having the sequence set forth in SEQ ID NO: 3 and three CDRs derived from the variable light chain region having the sequence set forth in SEQ ID NO: 4. In some embodiments, the anti-IFNβ antibody comprises an HCDR1 having the sequence set forth in SEQ ID NO: 5, an HCDR2 having the sequence set forth in SEQ ID NO: 6, an HCDR3 having the sequence set forth in SEQ ID NO: 7, an LCDR1 having the sequence set forth in SEQ ID NO: 8, an LCDR2 having the sequence set forth in SEQ ID NO: 9, and an LCDR3 having the sequence set forth in SEQ ID NO: 10. In some embodiments, the anti-IFNβ antibody comprises a variable heavy chain region having the sequence set forth in SEQ ID NO: 3 and a variable light chain region having the sequence set forth in SEQ ID NO: 4. In some embodiments, the anti-IFNβ antibody comprises a heavy chain having the sequence set forth in SEQ ID NO: 1 and a light chain having the sequence set forth in SEQ ID NO: 2, wherein the C-terminal lysine (K) of the heavy chain amino acid sequence of SEQ ID NO: 1 is optional. In some embodiments, the anti-IFNβ antibody comprises a VH encoded by the nucleic acid sequence of an insert in the vector deposited as CTI-AF1-VH with ATCC accession number PTA-122727, and a VL encoded by the nucleic acid sequence of an insert in the vector deposited as CTI-AF1-VL with ATCC accession number PTA-122726. In some embodiments, the antibody comprises a VH sequence encoded by an insert in a plasmid deposited with the ATCC with ATCC accession number PTA-122727. In some embodiments, the antibody comprises a VL sequence encoded by an insert in a plasmid deposited with the ATCC with ATCC accession number PTA-122726. In some embodiments, the anti-IFNβ antibody competes for binding with an anti-IFNβ antibody comprising a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4.In some embodiments, the anti-IFNβ antibody competes for binding with an antibody comprising a VH encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VH with ATCC accession number PTA-122727 and a VL encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VL with ATCC accession number PTA-122726.

[0374] The antibody may be present in the formulation at a concentration ranging from about 25 mg / ml to about 200 mg / ml, from about 40 mg / ml to 200 mg / ml, from about 50 mg / ml to about 175 mg / ml, or from about 60 mg / ml to about 150 mg / ml.

[0375] The antibody may be present in the formulation at a concentration of about 50 mg / mL. The antibody may be present in the formulation at a concentration of about 60 mg / mL. In some embodiments, the antibody is present in an amount of 50-70 mg / mL. In some embodiments, the antibody is present in an amount of about 60 mg / mL. Such concentrations are particularly suitable for IV dosing. In some embodiments, the antibody dosing regimen includes 600 mg of antibody every four weeks via IV injection. Such a 600 mg dosing regimen may preferably be provided at a concentration of 60 mg / mL to reduce viscosity and minimize drug product waste and dose storage. The formulations of the present disclosure are engineered to provide stable and safe dosing for both SC and IV formulations.

[0376] The antibody may be present in the formulation at a concentration of about 80 mg / ml. The antibody may be present in the formulation at a concentration of about 100 mg / ml. The antibody may be present in the formulation at a concentration of about 120 mg / ml. The antibody may be present in the formulation at a concentration of about 120 mg / ml to about 175 mg / ml. The antibody may be present in the formulation at a concentration of about 140 mg / ml. The antibody may be present in the formulation at a concentration of about 141 to about 154 mg / ml. The antibody may be present in the formulation at a concentration of about 150 mg / ml.

[0377] In some embodiments, the antibody is present in an amount of 140-160 mg / mL. The antibody may be present in the formulation at a concentration of about 140 mg / mL. In some embodiments, the antibody is present in an amount of 141-154 mg / mL. In some embodiments, the antibody is present in an amount of about 150 mg / mL. Such concentrations are particularly suitable for SC administration. In some embodiments, the antibody dosing regimen includes 600 mg of antibody administered weekly via SC injection. Such a 600 mg dosing regimen may preferably be provided at a concentration of 150 mg / mL to minimize local injection volume. The formulations of the present disclosure are engineered to provide stable and safe doses for both SC and IV formulations.

[0378] In some embodiments, the buffering agent (His or His-HCl) provides a formulation with a pH close to physiological pH to reduce the risk of pain or anaphylactic-like side effects upon injection, and also provides enhanced antibody stability and resistance to aggregation, oxidation, and fragmentation. In some embodiments, the buffering agent is His. In some embodiments, the buffering agent is His-HCl.

[0379] The concentration of the buffering agent can range from about 1 millimolar (mM) to about 100 mM, preferably from about 5 mM to about 50 mM, more preferably from about 10 mM to about 30 mM, and even more preferably from about 15 mM to about 25 mM. Preferably, the concentration of the buffering agent is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM. In some embodiments, the buffering agent is His buffer at a concentration of about 20 mM.

[0380] The concentration of the stabilizer can range from about 1 millimolar (mM) to about 100 mM. Preferably, the concentration of the stabilizer is about 10 mM to about 90 mM, more preferably about 25 mM to about 75 mM, and even more preferably about 40 mM to about 60 mM. Preferably, the concentration of the stabilizer is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM. In some embodiments, the stabilizer is present at a concentration of about 50 mM.

[0381] In some embodiments, the stabilizer is arginine or NaCL. In some embodiments, the stabilizer is arginine. In some embodiments, the stabilizer is arginine at a concentration of about 1 mM to about 100 mM. In some embodiments, the stabilizer is arginine at a concentration of about 25 mM to about 75 mM. In some embodiments, the stabilizer is arginine at a concentration of about 50 mM to about 60 mM. In some embodiments, the stabilizer is arginine at a concentration of about 50 mM.

[0382] In some embodiments, the polyol can have a molecular weight of less than about 600 kD (e.g., in the range of about 120 to about 400 kD) and contain multiple hydroxyl groups, including sugars (e.g., reducing and non-reducing sugars or mixtures thereof, sugars or carbohydrates), sugar alcohols, sugar acids, or salts or mixtures thereof. Examples of non-reducing sugars include, but are not limited to, sucrose, trehalose, and mixtures thereof. In some embodiments, the polyol is mannitol, trehalose, sorbitol, erythritol, isomalt, lactitol, maltitol, xylitol, glycerol, lactitol, propylene glycol, polyethylene glycol, inositol, or mixtures thereof. In other embodiments, the polyol can be, for example, without limitation, a monosaccharide, disaccharide, or polysaccharide, or a mixture of any of the foregoing. The sugar or carbohydrate may be, for example, without limitation, fructose, glucose, mannose, sucrose, sorbose, xylose, lactose, maltose, dextran, pullulan, dextrin, cyclodextrin, soluble starch, hydroxyethyl starch, water-soluble glucan, or a mixture thereof. The polyol may be sucrose or trehalose. The polyol may be trehalose. The polyol may be sucrose.

[0383] In some embodiments, the concentration of the polyol in the formulation ranges from about 1 mg / ml to about 300 mg / ml, about 1 mg / ml to about 200 mg / ml, or about 1 mg / ml to about 120 mg / ml. Preferably, the concentration of the polyol in the formulation is from about 10 mg / ml to about 100 mg / ml, about 20 mg / ml to about 70 mg / ml, or about 40 mg / ml to about 60 mg / ml. In some embodiments, the concentration of the polyol in the formulation is about 1 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, about 4.5 mg / ml, about 5 mg / ml, about 5.5 mg / ml, about 6 mg / ml, about 6.5 mg / ml, about 7 mg / ml, about 7.5 mg / ml, about 8 mg / ml, about 8.5 mg / ml, about 9 mg / ml, about 9.5 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13mg / ml, about 14mg / ml, about 15mg / ml, about 16mg / ml, about 17mg / ml, about 18mg / ml, about 19mg / ml, about 20mg / ml, about 21mg / ml, about 22mg / ml, about 23mg / ml, about 24mg / ml, about 25mg / ml, about 26mg / ml, about 27mg / ml, about 28mg / ml, about 29mg / ml, about 30mg / ml, about 31mg / ml, about 32mg / ml, about 33mg / ml, about 34mg / ml, about 35mg / ml, about 36mg / ml, about 37mg / ml, about 38mg / ml, about 39mg / ml, about 40mg / ml, about 41mg / ml, about 42mg / ml, about 43mg / ml, about 44mg / ml, about 45mg / ml, about 46mg / ml, about 47mg / ml, about 48mg / ml, about 49mg / ml, about 50mg / ml, about 51mg / ml, about 52mg / ml, about 53mg / ml, about 54mg / ml, about 55mg / ml, about 56mg / ml, about 57mg / ml, about 58mg / ml, Approximately 59mg / ml, approximately 60mg / ml, approximately 65mg / ml, approximately 70mg / ml, approximately 75mg / ml, approximately 80mg / ml, approximately 81mg / ml, approximately 82mg / ml, approximately 83mg / ml, approximately 84mg / ml, approximately 85mg / ml, approximately 86m g / ml, about 87 mg / ml, about 88 mg / ml, about 89 mg / ml, about 90 mg / ml, about 91 mg / ml, about 92 mg / ml, about 93 mg / ml, about 94 mg / ml, about 95 mg / ml, about 96 mg / ml, about 97 mg / ml,Approximately 98mg / ml, approximately 99mg / ml, approximately 100mg / ml, approximately 101mg / ml, approximately 102mg / ml, approximately 103mg / ml, approximately 104mg / ml, approximately 105mg / ml, approximately 106mg / ml, approximately 107mg / ml, approximately 108mg / ml, approximately 109mg / ml, approximately 110mg / ml, approximately 111m g / ml, approx. 112 mg / ml, approx. 113 mg / ml, approx. 114 mg / ml, approx. 115 mg / ml, approx. 116 mg / ml, approx. 117 mg / ml, approx. 118 mg / ml, approx. 119 mg / ml, approx. , about 125 mg / ml, about 126 mg / ml, about 127 mg / ml, about 128 mg / ml, about 129 mg / ml, about 130 mg / ml, about 131 mg / ml, about 132 mg / ml, about 133 mg / ml, about 134 mg / ml, about 135 mg / ml, about 136 mg / ml, about 137 mg / ml, about 138 mg / ml, about 139 mg / ml, about 140 mg / ml, about 141 mg / ml, about 142 mg / ml, about 143 mg / ml, about 144 mg / ml, about 145 mg / ml, about 146 mg / ml, about 147 mg / ml, about 148 mg / ml, about 149 mg / ml, or about 150 mg / ml. In some embodiments, the polyol is sucrose at a concentration of about 1 mg / ml to about 300 mg / ml, about 1 mg / ml to about 200 mg / ml, or about 1 mg / ml to about 100 mg / ml. In some embodiments, the polyol is sucrose at a concentration of about 10 mg / ml to about 90 mg / ml, about 20 mg / ml to about 80 mg / ml, or about 25 mg / ml to about 75 mg / ml. In some embodiments, the polyol is sucrose at a concentration of about 40 mg / ml to about 60 mg / ml. Preferably, the sucrose concentration is about 50 mg / ml.

[0384] When used in the present invention, surfactants can modify the surface tension of a liquid antibody formulation. In certain embodiments, surfactants reduce the surface tension of a liquid antibody formulation. In yet other embodiments, surfactants can contribute to improving the stability of any of the antibodies in the formulation. Surfactants can also reduce aggregation of the formulated antibody (e.g., during transportation and storage), minimize particle formation in the formulation, and / or reduce adsorption (e.g., adsorption to the container). For example, surfactants can also improve the stability of the antibody during and after freeze / thaw cycles. Surfactants include, but are not limited to, polysorbates, poloxamers, triton, sodium dodecyl sulfate, sodium lauryl sulfate, sodium octyl glycoside, lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, stearyl sulfobetaine, lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, stearyl sarcosine, linoleyl betaine, myristyl betaine, cetyl betaine, lauroamidopropyl betaine, cocamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmidopropyl betaine, isostearamidopropyl betaine, myristamidopropyl dimethylamine, palmidopropyl dimethylamine, isostearamidopropyl dimethylamine, sodium cocoyl methyl taurate, disodium oleyl methyl taurate, dihydroxypropyl PEG The surfactant may be, for example, but not limited to, polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85, PEG3350, or a mixture thereof. In some embodiments, the surfactant is polysorbate 80 (PS80).

[0385] The concentration of the surfactant is generally about 0.01 mg / ml to about 10 mg / ml, about 0.01 mg / ml to about 5.0 mg / ml, about 0.01 mg / ml to about 2.0 mg / ml, about 0.01 mg / ml to about 1.5 mg / ml, about 0.01 mg / ml to about 1.0 mg / ml, about 0.01 mg / ml to about 0.5 mg / ml, about 0.01 mg / ml to about 0.4 mg / ml, about 0.01 mg / ml to about 0.3mg / ml, about 0.01mg / ml to about 0.2mg / ml, about 0.01mg / ml to about 0.15mg / ml, about 0.01mg / ml to about 0.1mg / ml, about 0.01mg / ml to about 0.05mg / ml, about 0.1mg / ml to about 1mg / ml, about 0.1mg / ml to about 0.5mg / ml, or about 0.1mg / ml to about 0.3mg / ml. More preferably, the concentration of the surfactant is about 0.05 mg / ml, about 0.06 mg / ml, about 0.07 mg / ml, about 0.08 mg / ml, about 0.09 mg / ml, about 0.1 mg / ml, about 0.15 mg / ml, about 0.2 mg / ml, about 0.3 mg / ml, about 0.4 mg / ml, about 0.5 mg / ml, about 0.6 mg / ml, about 0.7 mg / ml, about 0.8 mg / ml, about 0.9 mg / ml, or about 1 mg / ml. In some embodiments, the polysorbate is polysorbate 80 at a concentration ranging from about 0.1 mg / ml to about 0.3 mg / ml. In some aspects, the surfactant is PS80 and is present in an amount of 0.2 mg / ml.

[0386] The formulation may further comprise a chelating agent. In the formulation of the present invention, the chelating agent can reduce the formation of reactive oxygen species, reduce the formation of acidic species (e.g., deamidation), reduce antibody aggregation, and / or reduce antibody fragmentation, and / or reduce antibody oxidation. For example, the chelating agent can be a multidentate ligand that forms at least one bond (e.g., covalent, ionic, or otherwise) with a metal ion and acts as a stabilizer, forming complexes with species that may normally promote instability.

[0387] In some embodiments, the chelating agent may be selected from the group consisting of aminopolycarboxylic acids, hydroxyaminocarboxylic acids, N-substituted glycines, 2-(2-amino-2-oxoethyl)aminoethanesulfonic acid (BES), deferoxamine (DEF), citric acid, niacinamide, and desoxycholic acid, and mixtures thereof. In some embodiments, the chelating agent may be ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), nitrilotriacetic acid (NTA), N-2-acetamido-2-iminodiacetic acid (ADA), bis(aminoethyl)glycol ether, N,N,N',N'-tetraacetic acid (EGTA), trans-diaminocyclohexanetetraacetic acid (DCTA), glutamic acid, and aspartic acid, N-hydroxyethyliminodiacetic acid (HIMDA), N,N-bis-hydroxyethylglycine (bicine), and N-(trihydroxymethylmethyl)glycine (tricine). ), glycylglycine, sodium desoxycholate, ethylenediamine, propylenediamine, diethylenetriamine, triethylenetetraamine (triene), edetate disodium dihydrate (or EDTA disodium dihydrate or EDTA disodium salt), oxalate sodium EDTA, maleate, citric acid, citric acid monohydrate, and trisodium citrate dihydrate, 8-hydroxyquinolate, amino acids, histidine, cysteine, methionine, peptides, polypeptides, and proteins, and mixtures thereof.

[0388] In some embodiments, the chelating agent is selected from the group consisting of salts of EDTA, including dipotassium edetate, disodium edetate, calcium disodium edetate, sodium edetate, trisodium edetate, and potassium edetate, and a suitable salt of deferoxamine (DEF) is deferoxamine mesylate (DFM) or a mixture thereof. The chelating agent used herein may be the free acid or free base form of the compound, or a salt form, and may also be anhydrous, solvated, or hydrated forms of the compound or the corresponding salt. The chelating agent may be EDTA.

[0389] The chelating agent may be present in an amount of 0.01 to 0.1 mg / ml.The chelating agent may be present in an amount of 0.02 to 0.08 mg / ml.

[0390] The chelating agent may be present in an amount of 0.05 mg / ml. The chelating agent may be EDTA, which may be present in an amount of 0.05 mg.mL.

[0391] According to some embodiments, the pH may be within the range of about pH 5.0 to about 6.6, preferably about pH 5.0 to 6.5 or about pH 5.0 to 6.0, and most preferably pH 5.2 to 5.8. The pH of the formulations of the present disclosure may be within a range selected from about any one of pH 5.2, 5.3, 5.4, 5.5, or 5.6 to about any one of pH 6.5, 6.4, 6.3, 6.2, 6.1, 6.0, 5.9, 5.8, or 5.7. In some embodiments, the pH may be selected from any pH value of about pH 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, and 6.5, preferably the pH is pH 5.8±0.5, and most preferably the pH is pH 5.8±0.3.

[0392] In some embodiments, the formulation may contain a preservative. Preferably, the preservative is selected from phenol, m-cresol, benzyl alcohol, benzalkonium chloride, benzalkonium chloride, phenoxyethanol, and methylparaben. The concentration of the preservative generally ranges from about 0.001 mg / ml to about 50 mg / ml, from about 0.005 mg / ml to about 15.0 mg / ml, from about 0.008 mg / ml to about 12.0 mg / ml, or from about 0.01 mg / ml to about 10.0 mg / ml. Preferably, the concentration of the preservative is about 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, about 0.4 mg / ml, about 0.5 mg / ml, about 0.6 mg / ml, about 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, about 1.0 mg / ml, 2.0 mg / ml, 3.0 mg / ml, about 4.0 mg / ml, about 5.0 mg / ml, about 6.0 mg / ml, about 7.0 mg / ml, 8.0 mg / ml, 9.0 mg / ml, about 9.1 mg / ml, about 9.2 mg / ml, 9.3 mg / ml, 9.4 mg / ml, 9.5 mg / ml, 9.6 mg / ml, 9.7 mg / ml, 9.8 mg / ml, 9.9 mg / ml, or 10.0 mg / ml. Most preferably, the concentration of the preservative is about 0.1 mg / ml or 9.0 mg / ml. In some embodiments, the formulation is preservative-free.

[0393] kit The present invention also provides kits containing any or all of the anti-IFNβ antibodies described herein. The kits of the present invention include one or more containers containing the anti-IFNβ antibodies described herein and instructions for use according to any of the methods of the present invention described herein. Generally, these instructions include instructions for administering the anti-IFNβ antibodies for the above-mentioned therapeutic treatment. In some embodiments, the kits are provided to provide single-dose administration units. In certain embodiments, the kits may include both a first container containing dried protein and a second container containing an aqueous formulation. In certain embodiments, kits containing single- and multi-chamber pre-filled syringes (e.g., liquid syringes and lyophilized syringes) are included.

[0394] Instructions for use of anti-IFNβ antibodies generally include information regarding dosage, dosing schedule, and route of administration for the intended treatment. The container may be a unit dose, a bulk package (e.g., a multi-dose package), or a sub-unit dose. The instructions provided in the kits of the invention are typically written instructions on a label or package insert (e.g., a paper strip included in the kit), although machine-readable instructions (e.g., instructions on a magnetic or optical storage disk) are also acceptable.

[0395] The kit of the present invention is contained in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., hermetically sealed Mylar or plastic bags), etc. Packages for use in combination with specific devices, such as inhalers, intranasal administration devices (e.g., spray devices), or injection devices, such as minipumps, are also contemplated. The kit may have a sterile access port (e.g., the container may be an intravenous bag or a vial with a stopper that can be punctured by a hypodermic needle). The container may also have a sterile access port (e.g., the container may be an intravenous bag or a vial with a stopper that can be punctured by a hypodermic needle). At least one active agent in the composition is an anti-IFN-beta antibody. The container may further contain a second pharmaceutically active agent.

[0396] The kit may provide additional components, such as buffering agents and instructional information. Typically, the kit comprises a container and a label or package insert on or associated with the container.

[0397] Deposit of biological materials Representative materials of the present invention were deposited on December 18, 2015, with the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, USA. Vector CTI-AF1-VH, with ATCC accession number PTA-122727, contains a DNA insert encoding the heavy chain variable region of antibody CTI-AF1, and vector CTI-AF1-VL, with ATCC accession number PTA-122726, contains a DNA insert encoding the light chain variable region of antibody CTI-AF1. These deposits were made under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure and the Regulations thereunder (Budapest Treaty), which ensures maintenance of viable cultures of the deposits for 30 years from the date of deposit. The deposit is being made available by ATCC under the terms of the Budapest Treaty and is subject to an agreement between Pfizer Inc. and ATCC, which assures that upon the issuance of any relevant U.S. patent or the publication of any U.S. or foreign patent application, whichever occurs first, the progeny of the cultures of the deposit will be available in perpetuity and indefinitely, and will also assure the availability of the progeny to those determined by the Director of the U.S. Patent and Trademark Office to be entitled pursuant to 35 USC Section 122 and the Director's regulations thereunder (37 CFR Section 1.14, including, inter alia, reference to 886 OG 638).

[0398] The owner of this application agrees that if a culture of the deposited material dies or is lost or destroyed when cultured under suitable conditions, the material will be promptly replaced with another identical one upon notice. The availability of the deposited material shall not be construed as a license under the authority of any government to practice the invention in contravention of the rights granted pursuant to its patent laws. [Example]

[0399] Example 1 Study design for assessing anti-IFN-β antibodies C0251002 is an ongoing, double-blind, placebo-controlled, multistage, multicenter Phase 2 study to assess the efficacy, safety, and tolerability of PF-06823859 in adult participants with moderate to severe DM. The study consisted of four stages. Stages 1, 2, and corrected Stage 2 included participants with skin-predominant DM, and Stage 3 included participants with muscle-predominant DM. Figures 1, 2, 3, and 4 show the initial design and planned sample size for each stage. The actual number of individuals randomized is provided in the text below.

[0400] Stage 1: Participants with skin lesions (CDASI-activity ≥ 14 at screening) who had failed at least one standard of care systemic treatment (e.g., corticosteroids) were randomized 2:1 to receive 600 mg of PF-06823859 or placebo. Study drug or placebo administration occurred on days 1, 4, and 8. The primary endpoint (CFB CDASI-A) was assessed at week 12.

[0401] Stage 2: Participants with skin involvement (CDASI-activity ≥ 14 at screening) were randomized in a 5:11:4 ratio to receive 600 mg of PF-06823859, 150 mg of PF-06823859, or placebo. Study drug or placebo administration occurred on days 1, 4, and 8 of the study. The primary endpoint (CFB CDASI-A) was assessed at week 12.

[0402] Complementary Stage 2: A fixed-sequence design was employed in Complementary Stage 2, allowing all study participants the opportunity to receive active medication during the treatment period. Participants were randomized in a 5:11:2:2 ratio to one of the following sequences: 600 mg PF-06823859 followed by placebo, 150 mg PF-06823859 followed by placebo, placebo followed by 600 mg PF-06823859, or placebo followed by 150 mg PF-06823859. Study drug or placebo (determined by treatment sequence) was administered on Day 1, Week 4, Week 8, Week 12, Week 16, and Week 20. The primary endpoint (CFB CDASI-A) was assessed at Week 12 of Complementary Stage 2. After the treatment period ended at 24 weeks, participants then entered a 4-month follow-up period or were transferred to the long-term extension study C0251008.

[0403] Stage 3: A fixed-sequence design was also used in Stage 3, with participants primarily with muscle disease randomized 1:1 to one of the following sequences: 600 mg PF-06823859 followed by placebo or placebo followed by 600 mg PF-06823859, with treatment switching at week 12. Inclusion criteria for muscle disease required subjects to meet one of the following two criteria: (1) MMT-8 of 136 / 150 or less and PhGA (VAS ≥ 3 cm on a 0-10 cm scale) or (2) the sum of PhGA, PtGA, and extramuscular global assessment ≥ 10 cm (each using a 0-10 cm VAS scale) and failure of at least two adequate courses of immunosuppressants or immunomodulators, including intravenous immunoglobulin (IVIG). Stable doses of immunosuppressants and immunomodulators, including IVIG, were permitted as concomitant medications. Administration of the study drug or placebo (determined by treatment sequence) occurred on Day 1, Week 4, Week 8, Week 12, Week 16, and Week 20. Secondary endpoints (TIS) were assessed longitudinally at Weeks 4, 8, and 12 (Week 12 was the critical time point). Other secondary muscle-related endpoints were MMT-8 and PtGA of myositis. After Week 12, the sequence was switched to the other treatment. After the treatment period ended at Week 24, participants entered a 4-month follow-up period or entered the long-term extension study C0251008.

[0404] Example 2 Selection of endpoints for assessing the efficacy of anti-IFN-β antibodies Primary endpoint 1. Stage 1: The primary objective was to assess the efficacy of PF-06823859 as measured by change from baseline (CFB) in CDASI-A score at week 12. Formal statistical testing was performed only in Stage 1. 2. Stage 2 and Corrected Stage 2: The primary objective was to assess the efficacy of PF-06823859, as measured by change from baseline in CDASI-A score at week 12 (CFB), in a pooled efficacy analysis of data from baseline to week 12 in Stage 1, Stage 2, and Corrected Stage 2. 3. Stage 3: The primary objective was to assess the safety and tolerability of PF-06823859 as measured by the occurrence of AEs, laboratory abnormalities, vital sign changes, and ECG findings.

[0405] Secondary endpoints Skin cohort (pooled data from stage 1, stage 2, and corrected stage 2) [weeks 0 to 12] 4. CFB and CFB percent of CDASI-A scores over time 5. Proportion of CDASI-A responders (CDASI-A CFB > 5, CDASI-A CFB percentage > 40%) at Week 12 6. Safety and tolerability as measured by the occurrence of AEs, laboratory abnormalities, changes in vital signs, and ECG findings Muscle cohort [Week 0 to Week 12] (Stage 3) 7. TIS 8. Muscle disease-related endpoints 1. Manual Muscle Testing (MMT-8) 2. Patient Global Assessment (PtGA) of Myositis 3. Physician Global Assessment (PhGA) for Myositis 4. Muscle Damage Biomarker: Creatine Kinase (CK) Levels

[0406] Statistical analysis populations and methods The primary analysis populations for safety and efficacy are as follows: 9. The full analysis set in Stage 1 (FAS1) includes all participants who received at least one dose of randomized treatment in Stage 1. 10. The full analysis set in stage 3 (FAS3) includes all participants who received at least one dose of randomized treatment in stage 3. 11. The Dermatologically Predominant Stage Pooled Complete Analysis Set (PFASS) includes all subjects who received at least one dose of randomized treatment in Stage 1, Stage 2, or Corrected Stage 2. 12. The Safety Analysis Set (SAS) includes all subjects who received at least one dose of randomized treatment. The subsets of the SAS specific to Stage 1, Stage 2, Corrected Stage 2, and Stage 3 are abbreviated as SAS1, SAS2, SASA2, and SAS3, respectively.

[0407] In the Stage 3 muscle cohort, TIS / CK at week 12 was assessed using an MMRM model with data from all post-baseline visits up to week 12. The model used TIS / CK as the outcome, with fixed effects for treatment, time (visit), and treatment-by-time, and an unstructured covariance matrix. CDASI-A / MMT-8 / PtGA / PhGA / CK CFB at week 12 was assessed using a LANCOVA model with data from all post-baseline visits up to week 12. The model used CDASI CFB as the outcome, with baseline value, treatment, time (visit), and treatment-by-time as covariates. An unstructured covariance matrix was used. In both MMRM and LANCOVA analyses (except for the comparison between change from baseline in CDASI-A at week 12 in Stage 1), p values ​​were exploratory and not adjusted for multiplicity of comparisons.

[0408] Example 3 result Study population, breakdown, and demographics Muscle Cohort A total of 18 participants with myopathy-predominant DM from five countries (USA, Poland, Hungary, Spain, and Germany) were randomized (treated). A total of 18 participants completed the 12-week treatment period, and only 6 (33.3%) completed the 24-week treatment period. Table 4 provides an overview of the breakdown of participants in Stage 3.

[0409] Skin cohort A total of 32, 9, and 16 participants with dermatosis-predominant DM from two countries (the United States and Hungary), respectively, were randomized (treated). Table 5 shows the number of participants randomized, who completed the 12-week treatment and follow-up periods, and who discontinued in Stage 1 and Stage 2. Table 6 shows the number of participants randomized, who completed the 24-week treatment period (and the 12-week period divided into the first and second halves) and follow-up periods, and who discontinued in Complementary Stage 2. The discontinuation rates over the first 12 weeks of Stage 1, Stage 2, Complementary Stage 2, and Stage 3 were 9.4%, 0%, 0%, and 0%, respectively.

[0410] [Table 4]

[0411] [Table 5]

[0412] [Table 6]

[0413] Demographic and baseline disease characteristics Muscle Cohort Baseline characteristics for each treatment sequence in Stage 3 were generally balanced and are summarized in Table 7. The majority of patients were Caucasian (89%) and female (72%), and the median disease duration was 2 years. Concurrent (ongoing) medications included oral steroids (66.7%), IVIG (38.9%), and immunosuppressants (e.g., MMF, AZA, MTX) (77.8%), either as monotherapy or in combination.

[0414] Skin cohort Baseline characteristics for each treatment sequence in Stage 1, Stage 2, and Corrected Stage 2 were generally balanced and are summarized in Tables 8 and 9. The majority of patients were Caucasian (90.6%, 100%, and 93.8% in Stage 1, Stage 2, and Corrected Stage 2, respectively) and female (90.6%, 100%, and 93.8%, respectively). In the overall population, 53%, 55.6%, and 50% of participants were continuing to use steroids in Stage 1, Stage 2, and Corrected Stage 2, respectively, while 12.5%, 44.4%, and 12.5% ​​were continuing to use IVIG.

[0415] [Table 7]

[0416] [Table 8]

[0417] [Table 9]

[0418] Total Improvement Score (TIS) The estimated mean TIS scores (90% CI) at weeks 4, 8, and 12, and the estimated difference in mean scores between the active and placebo arms, are presented in Table 10. The sample size for this study was not large enough to test for the presence of a treatment effect (a positive difference in mean TIS values ​​between the active and placebo treatment groups) at the week 12 visit. At week 12, the mean TIS score values ​​for PF-06823859 and placebo were 56.4 (41.4, 71.4) and 36.9 (22.0, 52.0). The high placebo effect may be attributable to ongoing concomitant medications, which included oral steroids (66.7%), IVIG (38.9%), and immunosuppressants (e.g., MMF, AZA, MTX) (77.8%), either alone or in combination.

[0419] [Table 10]

[0420] Figure 5 shows the estimated mean TIS in each treatment group from baseline to week 12 (left panel), and the difference in mean TIS scores between the active and placebo groups (right panel). Note that TIS scores assess change from baseline, not defined at baseline (which is equal to 0 for each subject), so the value at week 0 is shown for reference only. The mean global improvement score (TIS) tended to increase over time (weeks 0-12) without plateauing until week 12, indicating numerical superiority of PF-06823859 600 mg compared to placebo.

[0421] A sensitivity analysis was performed after removing the single efficacy observation for subjects taking prohibited medications. The estimated mean TIS values ​​at week 12 were 56.4 (90% CI = (41.8, 71.0)) and 35.6 (90% CI = (20.9, 50.3)) for PF-06823859 and placebo, respectively, with a placebo-adjusted difference of 20.8 (90% CI = (0.0, 41.5)), reaching nominal statistical significance for TIS (p = 0.0497). The sensitivity analysis also reached nominal significance.

[0422] These values ​​and the estimated treatment effect of 19.4 (-1.8, 40.7) are comparable to the estimated treatment effect in the Octagam (IVIG) study (26.8 (19.0, 34.6)).

[0423] Other secondary endpoints related to muscle function Figure 6 shows the estimated mean (90% CI) change from baseline in MMT-8 (higher scores indicate improvement). PF-06823859 600 mg demonstrates a trend toward an increase over time. The mean CFB values ​​achieved at week 12 were 21.2 (11.9, 30.6) and 11.7 (2.3, 21.0) in the PF-06823859 600 mg and placebo treatment arms, respectively. The estimated treatment effect (90% CI) was 9.6 (-3.8, 23.0). These values ​​are comparable to the estimated treatment effect of MMT-8 observed in the ProDERM study (11.2 (6.9, 15.5)).

[0424] Figure 7 shows the estimated mean (90% CI) change from baseline in the Patient Global Assessment of Myositis. Lower scores indicate improvement, and the range used in this plot is 0 to 100 (a 100mm VAS is equivalent to a 10cm VAS). PF-06823859 600mg shows a trend toward a decrease over time (weeks 0 to 12) without reaching a plateau at week 12. The mean CFB values ​​for the PF-06823859 600mg and placebo arms at week 12 were -46.2 (-58.6, -33.7) and -11.7 (-24.1, 0.77). The estimated difference in means is -34.5 (-52.7, -16.2). These values ​​numerically exceed the estimated treatment effect of PtGA observed in the ProDERM study (-11.0 (-18.7, -3.3)).

[0425] Figure 8 shows the estimated mean change from baseline (90% CI) in creatine kinase. Mean baseline CK for PF-06823859 600 mg and placebo was 321.2 and 227.9, respectively. On the CFB scale, PF-06823859 600 mg demonstrated clear separation from placebo over weeks 4 through 12, achieving -185.8 (-273.9, -97.6) at week 12, while placebo achieved -39.9 (-125.7, 46.0), a difference of -145.9 (-269.4, -22.4). This improvement in muscle enzymes in patients treated with PF-06823859 is consistent with clinical improvement in muscle function, MMT-8 scores, and PtGA for myositis. The ProDERM study showed no treatment effect on CK (estimate was 183.6 (-364.0,731,2)).

[0426] Exposure-response modeling (detailed below) suggests that continuation of dosing up to 24 weeks would meet the muscle and skin efficacy requirements in the TPP for DM with respect to TIS, CFB MMT-8, and CDASI-A responses.

[0427] Skin cohort The mean (standard deviation) baseline CDASI-A scores for Stage 1, Stage 2, and corrected Stage 2 were 32.7 (10.6), 30.8 (9.4), and 33.3 (11.3), respectively (Tables 8 and 9). The estimated CFB and placebo-adjusted differences in CDASI-A scores for participants receiving 600 mg PF-06823859, 150 mg PF-06823859, and placebo for the first 12 weeks in the pooled skin cohort at week 12 are presented in Table 11. Both doses of PF-06823859 were different from placebo at week 12, p<0.0001. Separation from placebo occurred as early as week 4, with no plateau until week 12. The estimated difference in the mean CFB (CDASI-A) at week 12 presented in Table 11 numerically exceeds the estimated CDASI-A treatment effect observed in the ProDERM study (-8.2 (-11.3, -5.3)).

[0428] [Table 11]

[0429] A sensitivity analysis was performed on the CDASI-A after removing one subject who took a prohibited medication in the 600 mg treatment arm in Stage 2. The sensitivity analysis was consistent with the primary analysis for mean CFB on the CDASI-A.

[0430] Figure 9 shows that the placebo and 600 mg dose estimates in the pooled sample are similar to the estimates based on the Stage 1 data. The efficacy estimates for the 150 mg and 600 mg treatment arms based on the pooled data are nearly identical.

[0431] Supporting analyses for the CDASI-A score using responder rates (Table 12) and percentage change from baseline (Table 13) were consistent with the primary analysis, demonstrating that both doses of PF-06823859 were differentially associated with placebo. A 4- or 5-point change in CDASI-A score represents a minimally clinically meaningful change (Anyanwu et al., 2015), and PF-06823859 achieved this reduction in CDASI-A score in more than 96% of subjects. For patients with a CDASI-A score greater than 14 points, a 40% change in CDASI-A score indicates a meaningful change in quality of life (Ahmed et al., 2020), and PF-06823859 achieved this reduction in CDASI-A score in more than 80% of subjects.

[0432] [Table 12]

[0433] At week 12, the mean CFB% on the CDASI-A for 150 mg and 600 mg PF-06823859 compared to placebo was -44% and -50%, respectively (compared to -35% at week 24 in the current targeted product profile).

[0434] [Table 13]

[0435] In summary, both doses of PF-06823859 (600 mg and 150 mg) met the primary efficacy endpoint for the skin cohort, as assessed by CDASI-A score. Although this study was not powered to assess efficacy for the muscle cohort, PF-06823859 600 mg was numerically superior to placebo across all muscle function endpoints (TIS, MMT-8, PtGA for myositis, and CK), with nominal statistical significance for PtGA and CK. Sensitivity analyses excluding single time points for placebo subjects receiving prohibited concomitant medications reached nominal statistical significance for TIS, CK, and PtGA. The 12-week efficacy results for CDASI-A were favorable, and the results for muscle-related endpoints (i.e., TIS, MMT-8, PtGA, CK) were comparable to or numerically better than the efficacy results obtained from the only approved treatment for DM (the ProDERM study).

[0436] Pharmacokinetics In DM patients with primarily dermatological disease who received 150 or 600 mg Q4W, there was an approximately 4-fold and 3.2-fold increase in AUC and Cmax, respectively (Table 14). The PK of PF-06823859 was similar in skin-predominant and muscle-predominant DM and was consistent with exposure in healthy participants in Phase 1 (Table 14). PK data were available for only two Stage 3 participants for analysis in Table 11.

[0437] [Table 14]

[0438] Mechanistic Biomarker Assessment: Percent Change from Baseline in IP-10 IP-10 decreased over time, with the mean percent reduction in IP-10 at week 12 (pooled analysis) being similar between patients with cutaneous-predominant DM who received either PF-06823859 150 mg (n=14, -70.25%) or 600 mg (n=27, -67.58%). The mean percent reduction in IP-10 for Stage 3 participants (n=9) who received placebo after active agent was -38.89%. The model predicted a median IP-10 level of 624.0 pg / mL (90% prediction interval 507.2, 763.6) at baseline for participants with cutaneous and muscle-predominant DM, which decreased to 298.7 (255.2, 349.1) and 255.9 (220.0, 304.2) pg / mL at week 12 for 150 and 600 mg PF-06823859 doses, respectively. There were no model-identifiable differences between cutaneous and muscle-predominant subjects.

[0439] Exposure-Response A preliminary model was developed to predict clinical endpoints in response to IFNb inhibition predicted from the PKPD model. A clinical trial simulation including participants from all stages was used to predict CDASI-A and TIS responses after 24 weeks of continuous treatment with PF-06823859 or placebo (Table 15). A key limitation of this model is the assumption that all endpoints have the same drug effect. This assumption was necessary given limitations in the small sample size, particularly for the muscle cohort. The preliminary model predicts that non-TIS endpoints, such as CDASI-A, will plateau at 12 weeks, and TIS endpoints will plateau at 14 weeks. Endpoint results are based on 300 trials in 100 participants (50 active, 50 placebo). TIS and MMT-8 results were based on stage 3 demographics, and CDASI-A results were based on participants from earlier stages. Point estimates from the model differ slightly from the observed summary responses but are generally consistent.

[0440] [Table 15]

[0441] immunogenicity Among patients with cutaneously predominant dermatosis, ADAs were reported in 3 of 22 (13.6%) and 1 of 5 (20%) patients in Stage 1 (600 mg IV) and Stage 2 (150 mg), respectively. No ADAs were reported in any of the other stages. The low incidence of drug-induced ADAs (3 / 22, Stage 1-600 mg) implies a lack of clinically relevant impact on safety / PK / PD or efficacy. Exploratory modeling of the effect of ADAs on PKPD parameters also did not suggest any impact. No events of anaphylaxis or concerning immunologically related clinical responses were reported.

[0442] safety The majority of TEAEs were mild and no dose-related association was observed. There were numerically higher numbers of TEAEs in the 150 mg group than in the 600 mg group and placebo. The most common TEAEs by SOC in this study were infections and infestations. The 600 mg dose group and placebo had numerically higher rates of infections and infestations (27.7% and 20%, respectively) compared with the 150 mg dose group (11.8%). There were three treatment-related mild infection cases in the PF-06823859 group compared with two cases in the placebo group across all stages. Four cases of positive SARS-CoV-2 testing were reported in the placebo group, and one case of suspected COVID-19 was reported in the PF-06823859 600 mg group. There were no cases of shingles or herpes simplex in this study.

[0443] Additional common TEAEs by SOC (Table 16) were laboratory tests (600 mg group = 19.1%, 150 mg group = 11.8%, placebo group = 22.2%), nervous system disorders (600 mg group = 14.9%, 150 mg group = 23.5%, placebo group = 13.3%), gastrointestinal disorders (600 mg group = 12.8%, 150 mg group = 11.8%, placebo group = 17.8%), and skin and subcutaneous tissue disorders (600 mg group = 14.9%, 150 mg group = 23.5%, placebo group = 13.3%).

[0444] Pharmacokinetics / Pharmacodynamics A population PKPD model was developed based on available PF-06823859 serum concentrations, IFNb levels, IP-10 levels, and gene signatures obtained from lesional and non-lesional skin and blood from C0251001 (if applicable) and C0251002. The most common (>5%) TEAEs by PT (preferred term) were headache (N=3, 17.6%) and arthralgia (N=2, 11.8%) for the 150 mg group (single cases for all other PTs). The most common PTs in the 600 mg group were headache (N=6, 12.8%) and upper respiratory tract infection (N=4, 8.5%). The most common PTs in the placebo group were headache (N = 5, 11.1%), pruritus (N = 5, 11.1%), positive SARS-CoV-2 test (N = 4, 8.9%), fatigue (N = 3, 6.7%), and sinusitis (N = 3, 6.7%).

[0445] [Table 16]

[0446] In Stage 1, two SAEs were reported in two participants receiving 600 mg of PF-06823859 and one SAE was reported in a participant receiving placebo. In Stage 2, there were no SAEs in participants receiving 600 mg of PF-06823859, 150 mg of PF-06823859, and placebo, respectively. In Correction Stage 2, there were no SAEs in those receiving either 600 mg or 150 mg of PF-06823859 or placebo during the first 12 weeks. In Stage 3, there were two SAEs in those receiving 600 mg of PF-06823859 during the first 12 weeks and no SAEs in those receiving placebo during the first 12 weeks.

[0447] There were no clinically significant changes from baseline in safety laboratory findings, ECGs, and vital signs after treatment with PF-06823859 150 mg and 600 mg.

[0448] In summary, in the skin cohorts of this study (stage 1, stage 2, and corrected stage 2), PF-06823859 administered once monthly for a total of three doses of 150 mg (total N = 10) and 600 mg (total N = 25) was generally well tolerated and safe in patients with DM.

[0449] In the stage 3 muscle cohort (total N=9), PF-06823859 administered once monthly for a total of three doses of 600 mg was also generally well tolerated and safe in patients with DM.

[0450] Example 4 Support for the possible efficacy of anti-IFNB in ​​systemic lupus erythematosus (SLE), cutaneous lupus, lupus nephritis, and other select diseases. To evaluate the potential of our anti-IFNB antibody, we analyzed up to three fully powered transcriptome datasets in each of the following indications: atopic dermatitis (AD), alopecia, cachexia, Crohn's disease, cutaneous lupus, idiopathic pulmonary fibrosis, NASH, ulcerative colitis, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), scleroderma, and vitiligo. The studies varied in the number of tissues and patient samples collected. A total of 3,880 samples from lesional and non-lesional tissues, as well as patients and healthy control subjects (for comparison), were analyzed. Detailed information for each dataset is shown in Table 10.

[0451] Indication data were processed by CytoReason according to best practices and presented as normalized per-sample expression values. A specific type 1 IFN signature was monitored in a dermatomyositis study of an anti-IFNB antibody (PF-06823859). The signature was defined based on the mean gene expression (log2 CPM, counts per million reads) of the following 10 genes proposed by Wong et al. (2012): RSAD2, IFIT1, IFI44L, IFI27, IFI44, CXCL10, IFI6, ISG15, CMPK2, and HERC5.

[0452] For each indication, we calculated this sample-specific type 1 IFN signature. We then applied a one-sided Wilcoxon statistical test to assess differences between lesional and non-lesional or diseased and healthy tissues. Details of each comparison are shown in Table 17. We also applied the same method to our internal clinical trial data of an anti-IFNB antibody (PF-06823859) in dermatomyositis patients.

[0453] A total of 50 comparisons were tested, and a threshold of 0.001 was considered significant after correction for multiple testing. To assess the strength of differences in type 1 IFN signatures, effects were estimated as the median difference between lesioned and nonlesional or diseased and healthy tissues, respectively. Confidence intervals were obtained via the Hodges-Lehmann estimator or normal approximation, depending on the availability of accurate p-values.

[0454] Figure 10 highlights all studies with significant differences based on the 10-gene type 1 IFN-γ signature. Specifically, systemic lupus erythematosus (SLE), cutaneous lupus, and psoriasis show significant differences comparable to those observed in the internal anti-IFN-γ dermatomyositis study.

[0455] Other indications that show significant differences in type 1 IFN signatures based on the signature are ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma.

[0456] Figure 11 shows the effect of our anti-IFNB antibody on the type 1 IFN signature in relation to all other evaluated indications and their differential type 1 IFN signature signals. For each treatment arm (150 mg dose, 600 mg dose, and placebo) in our dermatomyositis study, we evaluate the type 1 IFN signature in lesional skin samples at baseline (circles) and after treatment (triangles) compared with non-lesional skin samples at baseline. Placebo treatment results in minimal changes in the lesional type 1 IFN signature. In contrast, the 150 mg and 600 mg doses reduce the signature in lesional skin to or below the level of non-lesional skin at baseline.

[0457] Collectively, these data support the potential of the anti-IFNB antibody PF-06823859 in additional indications, particularly SLE, cutaneous lupus, and psoriasis, and demonstrate its potential as a precision medicine for ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma.

[0458] Similar results were found when these data were repeated with a set of 13 type I IFN-inducible genes disclosed in Greenberg et al., 2012 and Tabata et al., 2023: (IFI44L, IFI44, IFIT1, IFIT3, RSAD2, ISG15, HERC5, IFI6, OAS3, MX1, EPSTI1, CXCL10, and CMPK2).

[0459] [Table 17-1]

[0460] [Table 17-2]

[0461] Example 5 Feasibility study of IFNβ-PF formulations at 150mg / ml or higher The current drug product (DP) of anti-IFNβ PF-06823859 is formulated at 100 mg / ml in 20 mM histidine, 85 mg / ml sucrose, 0.05 mg / ml EDTA, and 0.2 mg / ml polysorbate 80 at pH 5.8. Development of a higher-concentration formulation is planned to facilitate drug delivery to patients and allow flexibility for both intravenous (IV) and subcutaneous (SC) administration. This study assessed the feasibility of developing a 150 mg / ml PF-06823859 (IFNβ) formulation and focused on strategies to reduce viscosity at higher concentrations (>150 mg / ml).

[0462] The effect on product viscosity and osmolality was evaluated by varying pH, protein concentration, arginine, sucrose, and sodium chloride. Additionally, the viscosity of the drug substance in histidine buffer and in the complete formulation at different protein concentrations was compared.

[0463] The effect of pH To test the effects of pH and concentration, anti-IFNβ (PF-06823859) samples were prepared in 20 mM histidine buffer at pH 5.8, 5.5, and 5.0 (no excipients added). The original anti-IFNβ (PF-06823859) drug substance (in 20 mM histidine buffer, pH 5.8) was buffer-exchanged to the target pH value, then concentrated to approximately 190 mg / ml and diluted with 20 mM histidine buffer at the targeted pH. The prepared samples were tested for viscosity and concentration, and the results are plotted in Figure 12A. The results show that varying the pH from 5.0 to 5.8 at concentrations from 100 mg / ml to 190 mg / ml did not significantly affect the viscosity of the samples.

[0464] To assess whether the excipients (sucrose, PS80, and EDTA) affect viscosity, the viscosity of anti-IFNβ (PF-06823859) in 20 mM histidine buffer (pH 5.8) was compared to that of anti-IFNβ (PF-06823859) in a formulation (20 mM histidine buffer, 85 mg / ml sucrose, 0.2 mg / ml PS80, and 0.05 mg / ml EDTA). The results showed no significant difference in viscosity between the buffer and the complete formulation (Figure 12B). The raw data used for Figures 12A and 12B are shown in Table 18.

[0465] [Table 18-1]

[0466] [Table 18-2]

[0467] [Table 18-3]

[0468] Effect of arginine on viscosity To minimize viscosity of the more concentrated formulations, arginine (150 mM) was added to anti-IFNβ (PF-06823859) in 20 mM histidine buffer with 85 mg / ml sucrose (pH 5.8 and 5.5) and 85 mg / ml trehalose (pH 5.0). Trehalose was selected for the pH 5.0 sample to avoid hydrolysis of sucrose at low pH. The data in Table 18 show that the viscosities of all 150 mg / ml samples at pH 5.8, 5.5, and 5.0 were 13.5 cp, 13.5 cp, and 14.2 cp, respectively. The viscosities of the 180 mg / ml samples with 150 mM arginine were 23.3 cp and 25.2 cp at pH 5.8 and 5.0, respectively. This data indicated that viscosity was not reduced by lowering the pH of the samples. However, 150 mM arginine significantly reduced the viscosity from 29.8 cp in the sample without arginine (00706600-0229-M13) to 23.3 cp (00706600-0241-M20). It was noted that the osmolality tested was greater than 700 mOsm for all samples except M22, which was 673 mOsm.

[0469] [Table 19]

[0470] To make subcutaneous (SC) injection of highly concentrated anti-IFNβ (PF-06823859) drug products in prefilled syringes more feasible, osmolality values ​​below 500 mOsm are preferred. To reduce osmolality, additional testing was performed on 150 mg / ml anti-IFNβ (PF-06823859) formulated in 20 mM histidine buffer with lower arginine concentrations (100 mM, 50 mM) and lower sugar concentrations (50 mg / ml). The test results are listed in Table 19 and plotted in Figure 13, and Figure 14 shows that lowering the arginine concentration increases viscosity, so maintaining arginine levels above 50 mM is preferred. For all samples, the tested osmolality was below 500 mOsm, which met the study requirements.

[0471] [Table 20]

[0472] Effect of sodium chloride (NaCl) on IFNb viscosity To minimize the viscosity of the formulation, NaCl (50 mM) was tested for anti-IFNβ (PF-06823859) in 20 mM histidine buffer (pH 6.0 and 6.5) with 50 mg / ml sucrose. These samples were compared with samples containing 50 mM arginine (pH 6.0 and 6.5) and samples containing neither arginine nor NaCl (pH 6.0 and 6.5). All samples contained 150 mg / ml of anti-IFNβ (PF-06823859). The prepared samples were tested for viscosity, osmolality, concentration, and pH, and the data are shown in Table 21.

[0473] Figure 14 shows a comparison between the use of 50 mM arginine, 50 mM NaCl, and neither at pH 6.0 and 6.5. The data shows that increasing the pH has no benefit to viscosity, and that arginine is more capable of reducing viscosity than NaCl for anti-IFNβ (PF-06823859).

[0474] [Table 21]

[0475] This study assessed the feasibility of developing an anti-IFNβ (PF-06823859) formulation at or above 150 mg / ml and investigated the effects of pH, salt, arginine, and sucrose on the viscosity and osmolality of the formulation. The data showed that varying the pH from 6.5 to 5.0 did not significantly affect the viscosity of the anti-IFNβ (PF-06823859) samples, and that both arginine and sodium chloride were comparable in their ability to reduce the viscosity of the formulation.

[0476] Arginine concentrations of 50-100 mM or higher imparted satisfactory viscosity and osmolality levels to the formulation. Therefore, a high-concentration formulation for further study is proposed as 150 mg / ml of anti-IFNβ (PF-06823859) in 20 mM histidine buffer, pH 5.8, containing 50-100 mM arginine and 50-85 mg / ml sucrose, along with 0.2 mg / ml polysorbate 80 (PS 80) and 0.05 mg / ml EDTA.

[0477] Example 6 Formulation candidate nomination study for anti-IFNβ (PF-06823859) 150mg / ml drug product To evaluate anti-IFNβ (PF-06823859) formulations, a stability study was conducted using 150 mg / ml of anti-IFNβ (PF-06823859) drug product and varying amounts of arginine and sucrose in the formulation. In addition to assessing the physical (aggregation, precipitation, denaturation) and chemical (oxidation) stability of anti-IFNβ (PF-06823859), the study evaluated the viscosity of the formulation to identify the most feasible high-concentration formulation compatible with both IV and SC administration. The formulations used in this study are provided in the Table.

[0478] [Table 22]

[0479] Anti-IFNβ (PF-06823859) samples were filtered through a 0.2 micron PES filter and 1 mL of drug product was filled into 2 mL vials, sealed, and stored at -20°C, 5°C, 25°C, and 40°C. Samples were tested according to the stability sample withdrawal schedule in 20. A subset of samples was further subjected to freeze-thaw and agitation stress. Product quality attributes were assessed using the analytical methods listed in Table 23.

[0480] [Table 23]

[0481] [Table 24]

[0482] No significant changes in appearance or pH were observed for all four formulations across all storage conditions. Regarding UV-Vis concentration, no trends were observed for any formulations; some increase in concentration was noted at 8 weeks, likely due to the solution not being mixed thoroughly before testing; values ​​returned to normal after 12 weeks. Regarding SEC data, there was no significant change in % monomer after 12 weeks at -20°C or 5°C, with a decrease of approximately 1% at 25°C and approximately 6% at 40°C for all four formulations. The observed changes were expected for a high concentration monoclonal antibody at storage temperatures of 25°C and 40°C. Similarly, for iCE, there was no significant change in % acidic species at -20°C or 5°C over 12 weeks of stability, with an increase of approximately 5-6% at 25°C and approximately 35-40% at 40°C. Trends were similar for all four formulations, and the observed changes were expected for a high concentration monoclonal antibody at 25 and 40°C storage conditions. The nrCGE assay showed no significant downward trend in IgG% or upward trend in fragments% at -20°C and 5°C. An increase in fragments and a decrease in IgG with increasing temperature were observed for most of the formulations tested at 25°C and 40°C, which is expected for a high concentration monoclonal antibody at higher temperatures. For rCGE, no clear trends were observed in IgG% or fragments%, and the trends were similar for all formulations tested in this study. Methionine oxidation data showed an increasing trend in oxidation levels with storage temperature and time across all four formulations. There were no significant differences in T Onset values ​​measured by DSC (differential scanning calorimetry) for all four formulations. Subvisible particle counts were assessed by both HIAC and MFI, and all samples tested met USP <787> Test requirements were met: no specific increase or trend in subvisible particles was observed across all conditions and formulations after 12 weeks of storage.

[0483] Viscosity and osmolality were measured and the results are shown in Table 25.

[0484] [Table 25]

[0485] Results from this study showed that there were no significant physical (aggregation, precipitation, denaturation) or chemical (oxidation) differences in the stability profiles of all four formulations tested for 12 weeks at the specified storage conditions (5°C, 25°C, 40°C, and -20°C). In addition, samples subjected to freeze-thaw and agitation stress showed no significant changes in attributes across all formulations. Because the stability profiles of each formulation were comparable, viscosity and osmolality levels were determining factors for formulation nomination. The most desirable viscosity and osmolality were achieved with 50 mg / ml sucrose and 50 mM arginine.

[0486] Therefore, formulation M02 (IFNb-PF 06823859 150 mg / ml* in 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / ml EDTA, 0.2 mg / ml polysorbate 80, pH 5.8) was determined as the most suitable formulation for both IV and SC administration (*range 141-154 mg / ml).

[0487] Further modeling identified formulation M02 (IFNb-PF 06823859 150 mg / ml in 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / ml EDTA, 0.2 mg / ml polysorbate 80, pH 5.8) as particularly suitable for SC administration (range 141-154 mg / ml), and formulation M03 (IFNb-PF 06823859 60 mg / ml in 20 mM histidine, 50 mg / ml sucrose, 50 mM arginine, 0.05 mg / L EDTA, 0.2 mg / ml polysorbate 80, pH 5.8) as particularly suitable for IV administration, which minimizes the risk of vial pooling leading to product dilution and is cheaper to manufacture.

[0488] Example 7 Population modeling analysis In this analysis, we described the PKPD of PF-06823859 in healthy volunteers and DM patients, characterized the relationships between various biomarkers, and developed a model to evaluate dosing options for future trials.

[0489] Study design The studies included in this analysis were a first-in-human (FIH) study in healthy volunteers and a PII / IIb study for treating DM. The FIH study included single and multiple ascending dose groups delivering PF-06823859 via intravenous (IV) or subcutaneous (SC) routes. The DM study was divided into three stages, the first two of which focused on skin-predominant DM, and the last focused on muscle-predominant DM. In Stage 1 (S1), subjects were randomized to placebo or 600 mg Q4W × 3 IV dosing; S2 added a 150 mg dose level as well as a placebo crossover; and S3 included a placebo crossover but no 150 mg dose level.

[0490] For the IFNβ assay, the lower limit of quantitation (LLOQ) was 10 pg / mL. The gene signature was based on the expression of 13 genes (including IP-10 and other proteins induced by type I interferons) and reported as the log2 of mean counts per million of gene expression, which was treated as unitless in the analysis.

[0491] Previous knowledge and modeling experience Non-compartmental analysis of PK data in healthy volunteers showed that exposure was linear with little inter-individual variability (IIV) in that population. Graphical analysis showed no evidence of target-mediated drug deposition (TMDD), which may be due to low target concentrations in healthy volunteers.

[0492] The previous investigational drug for DM was sifalimumab, a mAb against IFNα (Higgs et al., 2014). A PKPD model of this drug and disease state has been developed that incorporates mechanistic relationships between the drug, IFN, and gene signatures (Wang et al., 2013). AMG-811 is another mAb against the type I interferon IFNγ, which has a PKPD model linking the drug, IFN, and interferon gamma-inducible protein 10 (IP-10) (Chen et al., 2015). These models were used to inform the development of the model in this analysis.

[0493] Modeling: Software and Strategy Nonlinear mixed-effects modeling was performed in NONMEM version 7.5.0, with some use of PsN 5.2.6 to facilitate uncertainty estimation using sampling importance resampling (SIR). Additional processing was performed in R. Modeling was performed using ADVAN13 with NONMEM to solve differential equations. The fitting algorithm was FOCE, which considered interactions, allowing the use of M3. All observed data were log-transformed on both sides.

[0494] Because initial examination of the healthy volunteer and DM data did not suggest TMDD or any other influence of PD on PK, the model was fit in a sequential manner. An individual PK parameter (IPP) approach to sequential PKPD modeling was applied.

[0495] Pharmacokinetic model description A pharmacokinetic model was developed using minimal covariates (only fixed allometric constants) to address all investigated administration routes and identify any differences between subject types. A typical structural model for mAbs consists of two compartments, which can be treated as first-order if absorption is relatively uncomplicated. The model was constructed considering results obtained from healthy volunteers for initial estimates and PK in nonhuman primates. Parameterization used clearance and volume macroconstants, and observations were fitted in ng / mL (log-transformed). Because the PD models were fitted separately, random effects and absorption lag were tested for the SC route, although subject data for fitting estimates were limited.

[0496] Basic model description A basic PD model attempted to provide a semi-mechanistic explanation for IFNβ binding and its association with downstream PD observations, such as IP-10 and gene signatures (GS). The system describing this model is shown in Equation 1. While no TMDD was addressed in this model, IFNβ binding and clearance used a quasi-steady-state approximation. Both protein biomarkers in the absence of drug were modeled with a simple turnover model. The median drug concentration (Conc) was defined as the quasi-steady-state binding constant (K SS ) was used in combination to estimate the fraction of total IFNβ bound. Bound and unbound IFNβ were calculated as internalization (rate constant k int ) and the essential decomposition (rate constant k deg ) and free / unbound IFNβ was calculated using the rate k determined from the steady-state assumption using baseline IFNβ. syn Free IFNβ was synthesized at the rate k* in the absence of essential IFNβ. syn,ip in a linear fashion (E sl p Based on the synthesis rate of IP-10 (k syn,ip ) was modeled to increase. For IP-10, k* syn,ip is also determined from the steady state assumption, and k syn,ipare individual baseline IFNβ estimates and E sl p The second compartment of IP-10 also exhibits an equivalent translocation rate constant, k res,ip Tested with.

[0497]

number

[0498] Although an IFNβ-independent steady-state baseline was fitted for IP-10, the longitudinal changes in this biomarker observed in placebo subjects prompted the investigation of non-steady-state (non-SS) effects. Modeling non-SS took advantage of the fact that the system was treated as static in a drug-free steady state, and effects were modeled as different initial IP-10 concentrations that would be defined within that state. Multiplier terms were estimated to increase or decrease the initial IP-10 compartment concentration above or below steady-state levels, thereby changing IP-10 concentrations under placebo conditions back to steady state. Non-SS multipliers would also affect the active-treatment groups. Regarding GS, the lack of extensive longitudinal data and the small range of observations limited the complexity with which these endpoints could be modeled. Therefore, GS was added at the end of the basic model development based on the observation that observed IP-10 concentrations and blood and lesional skin GS scores shared a double-reciprocal linear relationship, with non-lesional skin GS being proportional to lesional GS. Therefore, all GS scores are linearly modeled from the predicted IP-10 concentrations, which is described by the following equation: where the intercept (INT) is common for GS in lesional skin and blood, but the slope (SLP) of lesional skin (SLP) is common for GS in lesional skin and blood. GSL ) is the slope of the blood GS (SLP GSB ) coefficient f L-B The proportion of lesional skin GS considered in non-lesional skin was calculated as the parameter Prop N-L Modeling with.

[0499]

number

[0500] Similar to the PK submodel, the PD model fitted observations and related parameters in typical units (pg / mL for IFNβ and IP-10, unitless for GS), so arbitrary unit conversions were performed within the model code. Within the model, drug and IFNβ concentrations were converted to nanomolar units using their accepted molecular weight values.

[0501] Development of a random effects model Random effects were used to model IIV and residual unexplained variance (RUV). For IIV, the individual parameters K i As shown below, 2 The fixed effect estimates θ K and the random effects estimate η i Random effects were modeled as log-normally distributed with respect to the fixed effect parameters using

[0000] . In this model, the IIV parameters were mu-referenced to enhance stability, and any transformations required to preserve the reference were done in the code so that this construction remains valid. K i =θ K ·exp(η i ) (3)

[0502] RUV is modeled using additive and / or proportional variability, which is used to estimate the RUV for each individual observation at any given time (Obs for individual i at time j). i j ) and calculate the standard deviation (W) of the predicted value (Pre i j ) was obtained. A first-order approximation was applied to the additive RUV of the log-transformed data. In the following equation, Obs i j is the modeled dependent variable rather than the actual observation. For M3, the dependent variable was the likelihood (Like) that the predicted observation is less than the LLOQ, estimated by the cumulative distribution function Φ (two-sided transformation).

[0503]

number

[0504] The inclusion of random effects is guided by model diagnostics, and the retention of the random effect parameters is guided by parameter identifiability and shrinkage. Due to the compelling need to test random effects for certain parameters that may not be identifiable for many individuals (i.e., for these individuals, the empirical Bayes estimate (EBE) would be zero), shrinkage was calculated based only on non-zero values. Very low (<10-6) non-zero values ​​were also included in the shrinkage estimates.

[0505] Inclusion of covariates and development of the final model Covariates were generally added using a stepwise procedure. This process was followed by additional empirical analysis if indicated by clearly observable or theoretical differences in parameter values ​​between healthy and DM subjects, such as those affecting baseline for IFNβ, IP-10, and non-SS effects. Due to the small number of patients, extensive mapping of covariates was not possible, and we primarily considered those that showed differences between DM and healthy subjects. Covariates were typically included in the forward step with an α of 0.01 and retained in the backward step with an α of 0.001. Percent change in the standard deviation of the IIV estimates was also considered as an additional check on covariate inclusion.

[0506] For baseline, the baseline and IIV multipliers (Mb and Mv) of healthy subjects were simultaneously applied to DM subjects as shown below, considering two degrees of freedom in the likelihood ratio test. All of these covariate pairs were advanced for inclusion in a stepwise fashion, as controlling for key differences was expected to facilitate testing of other potential covariates. Backward elimination of multipliers involved sharing multipliers between baselines. Mb dm =exp(θdm ) or 1+θ dm Mv dm =θ var,dm Base hv =θ base Base i,hv =Base hv ·exp(η i ) Base i,dm =Mb dm Base hv ·exp(Mv dm η i ) (5)

[0507] Other covariates examined were modeled in a more standard manner: categorical covariates were fitted as percentage changes relative to typical parameter values, and continuous covariates were modeled using power relationships normalized to median or standard center estimates.

[0508] outliers Observations were suspected to be outliers if they had a conditional weighted residual (CWRES) greater than 6 or a normalized prediction distribution error (NPDE) greater than 2. Outliers were removed if they were found to be influential in parameter estimation (existing outliers differ by more than 10%).

[0509] Assessing model adequacy (goodness of fit) Standard diagnostic plots and indices were examined to assess goodness of fit. These plots included population and individual prediction plots, as well as distributions, correlations, and trends of EBE and residuals. Condition numbers were used to assess collinearity, with an upper bound set at 400. Where feasible, random effects were included only if shrinkage was less than 30%. Minimal bootstrap was used for model stability assessment. For uncertainty estimation, SIR was used with 1.2-fold inflation and default iteration and sample / resample ratios in PsN 5.2.6.

[0510] Evaluation (verification) of model prediction performance Predictive performance was assessed by visual predictive performance (VPC). Each VPC was associated with 1,000 simulated data sets. For PK, these were stratified by first dose, last dose (treated as steady state), and subject type. For IFNβ and IP-10, observations were stratified by subject type (for IFNβ), and BLQ% predicted was treated as a categorical endpoint. Gene signatures were stratified by site for longitudinal observation. Both skin gene signatures were evaluated together by the VPC of gene set improvement (GSI), defined below. This index normalizes the reduction in lesional (l) skin GS from baseline to the end of the study by the difference between non-lesional (nl) and lesional skin at baseline (the mean was used to account for the absence of non-lesional skin GS).

[0511] simulation Simulations were based on 500 simulated trials, each containing 100 simulated subjects, unless otherwise indicated. The following simulations were performed:

[0512]

number

[0513] The simulated adolescent population was obtained by weight sampling for children aged 12 to 17 years based on CDC weight table LMS parameters. The primary goal of the adolescent simulation was to estimate the effect of weight cutoffs and weight-based dosing on exposure. Therefore, for various weight cutoffs for exclusion, the distribution of steady-state AUC (AUCτ) and maximum concentration (Cmax) was considered, with the ratio of all simulated subjects to that of a 70 kg DM subject. In addition, given the cutoff range for dose type (lower receiving weight-based dosing and higher receiving the fixed adult dose), the percentage of subjects with a ratio greater than 2 was also considered.

[0514] result Observed data Although the total dataset represented a range of demographics, there were disparities between healthy and DM subjects that limited the investigation of model covariates. Baseline IFNβ was absent or BLQ in nearly all subjects. Where comparisons were possible, PK appeared to be similar between healthy and DM subjects. In DM, IFNβ response was associated with high variability, with most placebo recipients having no measurable levels. IFNβ concentrations were absent for S2 and S3 at the time of analysis. There was a clear placebo effect for IP-10 (levels decreasing over time), and there was overlap in response between 150 mg and 600 mg.

[0515] Pharmacokinetic model results Absorption was modeled with some complexity given the limited sample contributing data, but it was useful to limit the amount of information explained using a Vc random effect, which was reduced by including a random effect for the first-order absorption rate constant (ka); absorption lag had minimal impact on the random effect (however, estimates increased when introduced without a random effect for ka), but substantially improved the Akaike information criterion (AIC). Similarly, allometric constants did not significantly reduce the AIC or random effect estimates, but they were included in the final model because they are fixed and provide information for extrapolating to different ages. Antidrug antibodies were also evaluated as a time-varying covariate for CL, but did not show any significant effects that warranted their inclusion.

[0516] The final PK model parameters, including uncertainty estimates due to SIR, are listed in Table 26. The model demonstrated reasonable diagnostics and did not show any strong trends indicating the need for additional covariates or concurrent PKPD models.

[0517] [Table 26]

[0518] Base model results The basic PKPD model was able to capture all of the endpoints of interest in good condition, but the parameter uncertainty and IIV were high (Table 27). The high uncertainty is partly attributable to the large number of BLQ observations, and the distribution of EBEs indicates that the IIVs of both IFNβ and IP-10 appear to be bimodal. The model was moderately stable, with 67% successful minimization with limited bootstrapping (N=200).

[0519] [Table 27]

[0520] Final model results The final model showed improved parameter precision over the base model, with high IIV parameters mostly explained by the addition of covariates (Table 28). The non-SS fixed effects predict that the average healthy volunteer will experience an increase in IP-10 over the course of the study, and the average DM patient will experience a decrease in IP-10 (both under placebo conditions), rather than the typical decrease in any given subject predicted by the base model.

[0521] [Table 28]

[0522] Predictive performance of the final model VPC shows good predictive performance for PK (Figure 15) and pharmacodynamics (PD) in both healthy volunteers and patients. For IFNβ, variability is captured well, and BLQ% over time is well predicted within a narrow distribution (Figure 16). While the early trend in IP-10 is not fully captured by the model, the overall time course at all dose levels is captured within the predicted variability. GS and GSI were well predicted, but baseline lesional skin GS was underpredicted for the 600 mg group.

[0523] simulation Simulations show saturation of IFNβ binding at the dose levels studied in DM subjects (Figure 17). According to Figure 18, placebo or baseline IFNβ concentrations are unlikely to be observable for DM subjects at an LLOQ of 10 pg / mL; this figure also shows overlap between other PD endpoints, with a slight point effect benefit at 600 mg. Biomarkers are predicted to return to steady state within one year, although long-term extension studies are required to validate this prediction.

[0524] Ranged simulations show higher PD responses with increasing dose and dosing frequency, and a mixed effect with increasing frequency. While there is significant overlap between these measures, it can be seen that total cumulative dose and exposure still have a proportional association with significant biomarker responses up to 1200 mg (Figure 19). There is a trend for frequency to be lower than that present for dose and dosing frequency.

[0525] Youth Simulation Adolescent dosing supports a 40 kg weight cutoff for either exclusion or weight-based dosing (Figure 20), as this cutoff is associated with fewer than 10% of subjects predicted to have a two-fold higher exposure than adults. Using fixed-dose dosing across all possible weight ranges (down to the simulation of 22 kg), fewer than 20% of patients are predicted to have a two-fold higher exposure.

[0526] Consideration The model developed herein provides an efficient and flexible description of the PK and PD of PF-06823859. The drug and target are modeled using a standard quasi-steady-state approximation, and other biomarkers are modeled as downstream effects of IFNβ concentration, which has the advantage of being a semi-mechanistic approach. Therefore, this model can infer IFNβ concentration in the absence of IFNβ concentration, as long as the subject has at least available IP-10 concentration, which can also predict GS (at least for cutaneous predominant DM).

[0527] The ability to account for both healthy nonpatients and DM patients is valuable given the rarity of the disease state being investigated. While demographic disparities limited extensive covariate evaluation in this analysis, additional healthy subject data from other Phase I trials, as well as data from C0251002, the expansion study, and the end of Phase III, may facilitate these investigations. From this analysis, IV PK was found to be highly consistent across disease states, and major PD differences were addressed; there was no strong signal for other demographic effects, although laboratory-based values, such as creatinine clearance and baseline albumin, may require further analysis. Finally, immunogenicity does not appear to have a strong effect or high incidence, but this may be limited to these subject types and / or studies and will warrant further investigation as additional data become available.

[0528] The model was used to inform dosing decisions for a Phase III trial, predicting that inclusion criteria will be expanded to adolescents and other patients with non-DM IIM. Simulations show that although there is clear saturation of IFNβ coverage even at low doses, key biomarker responses are predicted to continue improving with doses up to 1200 mg. They also show low sensitivity to frequency, with two doses recording similar point effect responses compared with three doses. Ultimately, these small differences will need to be reflected in clinical responses, but they could help justify lower frequency and fewer dosing doses if appropriate for future trials. Simulations in adolescents support a 40 kg weight cutoff; however, pharmacokinetic sampling in this population will be essential to verify the accuracy of these predictions. For adolescents, the threshold assessment was based on limiting exposure to a two-fold increase; the no-observed-adverse-event level (NOAEL) in cynomolgus monkeys was 25,600,000 ng / mL for Cmax and 2,430,000,000 ng h / mL for AUCτ, approximately 100- and 15-fold lower than this limit in adults, given the predicted median Cmax and AUCτ.

[0529] A limitation of this analysis is that a wide range of non-existent IFNβ concentrations were inferred from those that could be identified, which may have biased predictions toward skin-predominant DM. There is little literature describing the dynamic differences that may influence PKPD differences between skin-predominant and classic (or muscle-predominant) DM. While recent analyses in subsets of DM patients indicate that muscle-predominant subjects may have higher IP-10 expression (among other chemokines / cytokines), the model and observations of the few S3 subjects do not reflect this trend.

[0530] conclusion -We developed a model that semi-mechanistically describes the PKPD of IFNβ in healthy subjects and DM patients, while simultaneously addressing key differences between subject types. Biomarkers can be treated as correlated in response to treatment. Non-target biomarkers were modeled as dependent on IFNβ in both linear (GS) and non-linear (IP-10) responses. The effects of dose, frequency, and dosing frequency were simulated using multiple options available for consideration, and adolescent dosing simulations supported weight-based dosing or a 40 kg exclusion cutoff. Fixed dosing at lower adolescent weights is not predicted to increase exposure by more than fourfold in a 70 kg patient with DM.

[0531] Example 8 Population PK model A population pharmacokinetic (popPK) model was developed using data from a healthy volunteer study (C0251001) and a phase 2 patient study (C0251002). Nonlinear mixed-effects modeling was performed in NONMEM version 7.5.0, with PsN 5.2.6 used to facilitate uncertainty estimation using sampling importance resampling (SIR). Additional processing was performed in R. Modeling was performed using differential equations using ADVAN13 in NONMEM. The fitting algorithm was FOCE / Laplacian, accounting for interactions. All observed data were log-transformed on both sides.

[0532] The popPK model was developed using minimal covariates (only fixed allometric constants) and consists of two compartments with first-order absorption, representing the typical structure of mAbs (Ryman). The model was constructed considering results obtained from healthy volunteers for initial estimates and PK in non-human primates. Parameterization used clearance and volume macroconstants, and observations were fitted in ng / mL (log-transformed). The equations used to describe the final popPK model and the final estimates are as follows:

[0533]

number

[0534] Principles of subcutaneous medication The popPK model also estimated subcutaneous bioavailability based on data from six healthy volunteers who received the SC formulation in C0251001. The estimated subcutaneous bioavailability was 73.1%. Using the final popPK model, 100 studies were simulated, with 100 subjects per study and a 24-week follow-up. Various weekly subcutaneous dosing regimens were tested to match the PK exposure of the 600 mg IV Q4W dose. As shown in Figure 21 below, trough agreement is achieved between the SC dose and the 600 mg IV Q4W dose selected for the single pivotal study at the 140 mg SC weekly dose. As shown in Figure 22 below, agreement in AUC or Coverage (Cave) is achieved between the SC dose and the 600 mg IV Q4W dose at the 225 mg SC weekly dose.

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Claims

1. A method for treating a patient having one or more conditions associated with IFNβ expression in the patient, comprising administering to the patient an anti-IFNβ antibody in a dosing regimen sufficient to improve signs and symptoms of the one or more conditions by at least four weeks after initiation of treatment with the anti-IFNβ antibody, wherein the dosing regimen comprises multiple individual doses separated by at least one week.

2. 2. The method of claim 1, wherein the condition is one or more conditions selected from the group consisting of IIM, dermatomyositis, polymyositis, inclusion body myositis, SLE, cutaneous lupus, psoriasis, ulcerative colitis, Crohn's disease, rheumatoid arthritis, atopic dermatitis, and scleroderma.

3. 3. The method of any one of claims 1 to 2, wherein the condition is one or more conditions selected from the group consisting of dermatomyositis, polymyositis, inclusion body myositis, juvenile dermatomyositis, SLE, and cutaneous lupus.

4. 4. The method of any one of claims 1 to 3, wherein the condition is one or more conditions selected from the group consisting of dermatomyositis, polymyositis, inclusion body myositis, and juvenile dermatomyositis.

5. 5. The method of any one of claims 1 to 4, wherein one or more of the individual doses is an amount within a range having a lower limit selected from the group consisting of 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, and 600 mg and an upper limit selected from the group consisting of 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, and 1000 mg.

6. 6. The method of any one of claims 1 to 5, wherein one or more of the individual doses is between 140 mg and 600 mg.

7. 7. The method of any one of claims 1 to 6, wherein each individual dose is 140 mg.

8. 8. The method of any one of claims 1 to 7, wherein each dose is 150 mg.

9. 9. The method of any one of claims 1 to 8, wherein the individual doses are separated by one week from each other.

10. 10. The method of any one of claims 1 to 9, wherein each dose is administered by subcutaneous injection.

11. 7. The method of any one of claims 1 to 6, wherein each individual dose is 500 to 700 mg.

12. 11. The method of claim 10, wherein each dose is 600 mg.

13. 12. The method of any one of claims 10 to 11, wherein the individual doses are separated by 4 weeks from each other.

14. 14. The method of any one of claims 11 to 13, wherein each dose is by intravenous injection.

15. 3. The method of any one of claims 1-2, wherein the dosing regimen is continued for a period of time selected from the group consisting of at least 4 weeks, 1 month, 8 weeks, 2 months, 12 weeks, 3 months, 16 weeks, 4 months, 20 weeks, 5 months, 24 weeks, 6 months, and 26 weeks.

16. An improvement in signs or symptoms compared to placebo is characterized as a clinical response, and a clinical response is characterized by: (i) change from baseline in manual muscle testing (MMT-8) score greater than 0; (ii) an improvement in Total Improvement Score (TIS) greater than 0; (iii) change from baseline in patient global assessment score less than 0; (iv) an improvement in absolute muscle enzyme creatine kinase greater than 0; and (v) Change from baseline in Cutaneous Dermatomyositis Disease Area and Severity Index (CDASI-A) greater than 0 6. The method according to any one of claims 1 to 5, characterized by one or more measures selected from the group consisting of:

17. 17. The method of any one of claims 1 to 16, wherein the patient has previously been treated with at least one other medication selected from the group consisting of corticosteroids, IVIG, and immunomodulatory and immunosuppressive drugs, and optionally selected from the group consisting of hydroxychloroquine, azathioprine, mycophenolate mofetil, and methotrexate.

18. 18. The method of any one of claims 1-17, wherein the patient shows a clinical response or experiences signs or symptoms after a period selected from the group consisting of 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, and 26 weeks from the start of treatment.

19. The anti-IFNβ antibody (i) an isolated antibody comprising three CDRs derived from a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and three CDRs derived from a variable light chain region having the sequence set forth in SEQ ID NO:4; (ii) an isolated antibody comprising an HCDR1 having the sequence set forth in SEQ ID NO:5, an HCDR2 having the sequence set forth in SEQ ID NO:6, an HCDR3 having the sequence set forth in SEQ ID NO:7, an LCDR1 having the sequence set forth in SEQ ID NO:8, an LCDR2 having the sequence set forth in SEQ ID NO:9, and an LCDR3 having the sequence set forth in SEQ ID NO:10; (iii) an isolated antibody comprising a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4; (iv) an isolated antibody comprising a heavy chain having the sequence set forth in SEQ ID NO: 1 and a light chain having the sequence set forth in SEQ ID NO: 2, wherein the C-terminal lysine (K) of the heavy chain amino acid sequence of SEQ ID NO: 1 is optional; (v) an isolated antibody comprising a VH encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VH having ATCC accession number PTA-122727, and a VL encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VL having ATCC accession number PTA-122726; (vi) an isolated antibody comprising a VH sequence encoded by an insert in a plasmid having ATCC accession number PTA-122727 deposited with the ATCC; (vii) an isolated antibody comprising a VL sequence encoded by an insert in a plasmid having ATCC accession number PTA-122726 deposited with the ATCC; (viii) an isolated antibody that competes for binding with an anti-IFNβ antibody comprising a variable heavy chain region having the sequence set forth in SEQ ID NO:3 and a variable light chain region having the sequence set forth in SEQ ID NO:4; and (ix) an isolated antibody that competes for binding with an antibody comprising a VH encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VH having ATCC accession number PTA-122727, and a VL encoded by the nucleic acid sequence of the insert of the vector deposited as CTI-AF1-VL having ATCC accession number PTA-122726; 19. The method of any one of claims 1 to 18, selected from the group consisting of:

20. The antibody is provided in an aqueous formulation, the aqueous formulation comprising: (i) an anti-IFNβ antibody at a concentration of 25 mg / mL to 200 mg.mL; (ii) histidine or His-HCl at a concentration of 10-50 mM; (iii) arginine or NaCl in an amount of 20 to 150 mM; (iv) sucrose or trehalose in an amount of 20 mg / ml to 85 mg / ml; (v) optionally comprising a chelating agent; (vi) a pH of between pH 5.0 and pH 6.5; 20. The method of any one of claims 1 to 19.

21. 21. Use of an anti-IFNβ antibody for the preparation of a medicament for the method of treatment according to any one of claims 1 to 20.

22. 21. An anti-IFNβ antibody for use in the method of any one of claims 1 to 20.

23. 21. Use of an anti-IFN beta antibody in the preparation of a medicament for treating a patient according to the method of any one of claims 1 to 20.

24. 1. An aqueous formulation comprising: (i) an anti-IFNβ antibody at a concentration of 25 mg / mL to 200 mg.mL; (ii) histidine or His-HCl at a concentration of 10-50 mM; (iii) arginine or NaCl in an amount of 20 to 150 mM; (iv) sucrose or trehalose in an amount of 20 mg / ml to 85 mg / ml; (v) optionally comprising a chelating agent; (vi) a pH of between pH 5.0 and pH 6.5; Aqueous formulation.

25. 21. An aqueous formulation for use in the method of any one of claims 1 to 20.

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