Methods of treating autoimmune diseases, such as primary immune thrombocytopenia (ITP), solid organ transplant rejection, graft-related disease, Pemphigus vulgaris, systemic sclerosis, and myasthenia gravis using antibody polypeptides that specifically bind human CD40L are provided. The antibody polypeptides do not activate platelets. The methods may comprise at least one administration cycle comprising one dose of the antibody polypeptide. The dose may be administered intravenously at a dose from about 75 mg to about 1500 mg. The method normalizes platelet counts in the human patient.

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation application of U.S. patent Ser. No. 16/795,823 filed Feb. 20, 2020, which is a continuation of U.S. patent Ser. No. 15/510,507 filed Mar. 10, 2017, which is a 35 U.S.C. § 371 National Stage patent application of International Application PCT/US2015/049338, filed Sep. 10, 2015, which claims priority to U.S. Provisional Application Ser. No. 62/048,459, filed Sep. 10, 2014; the entire contents of which are incorporated herein by reference.

REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] Incorporated herein by reference in its entirety is a Sequence Listing entitled, 12376-US-PCT_20230119_May1.xml, comprising SEQ ID NO:1 through SEQ ID NO:11, which include nucleic acid and/or amino acid sequences disclosed herein. The Sequence Listing has been submitted herein via Patent Center, and thus constitutes both the paper and computer readable form thereof. The Sequence Listing was first created using WIPO ST.26, which was submitted electronically via Patent Center. Said Sequence Listing XML copy, created on May 1, 2023 and is named 12376-US-PCT_20230119_May1.xml and is 9 KB in size.

TECHNICAL FIELD

[0003] Methods of treating autoimmune diseases, such as primary immune thrombocytopenia, solid organ transplant rejection, graft-related disease, Pemphigus vulgaris, systemic sclerosis, and myasthenia gravis using antibody polypeptides that specifically bind human CD40L are provided.

BACKGROUND

[0004] Idiopathic thrombocytopenic purpura (ITP), also known as primary immune thrombocytopenia, is an autoimmune disorder characterized by isolated thrombocytopenia (peripheral blood platelet count less than 100×10.sup.9/L) in the absence of other causes or disorders that may be associated with thrombocytopenia. Rodeghiero et al., Blood, 113(11):2386-2393 (2009). The underlying pathophysiology of ITP has traditionally been attributed to increased rates of destruction of antibody-coated platelets. In addition, impaired platelet production has emerged as an important mechanism contributing to the thrombocytopenia of ITP. The clinical manifestations of ITP are highly variable with a spectrum ranging from no symptoms to catastrophic hemorrhage. Symptoms are largely, though not exclusively, related to platelet count. Rodeghiero et al., Blood, 113(11):2386-2393 (2009).

[0005] The first-line treatment of patients presenting with ITP comprises oral or intravenous corticosteroids (prednisone is steroid most frequently used). Following initial response, relapse is common when the dose is reduced. George et al., Blood, 88(1):3-40 (1996). Long-term responses are seen in only around 20-30% patients. Provan et al., Blood, 115:168-186 (2010). Early management of those unresponsive to steroids includes intravenous immunoglobulins (IVIg), associated with response in about 80% cases. Newman et al., Br. J. Haematol., 112(4):1076-1078 (2001). However, the platelet counts often drift back to pre-treatment levels within 3-4 weeks. Treatment of patients with chronic ITP who fail first-line therapy is extremely challenging. Provan et al., Blood, 115:168-186 (2010). Chances of inducing a durable and complete response are low in these patients; therefore, the goals of therapy are to provide a “safe” platelet count to prevent major (including potentially fatal) bleeding while minimizing treatment-related side effects. Splenectomy is the only curative treatment modality and two-thirds of patients who undergo splenectomy will achieve a normal platelet count which is often sustained with no additional therapy. Ghanima et al., Blood, 120(5):960-969 (2012). There is no consensus regarding the optimal time for performing splenectomy, or for predicting response and the long-term effectiveness of splenectomy. In about 30% of patients, splenectomy fails to induce a satisfactory response and these patients require additional treatment. Cuker et al., Hematol. Am. Soc. Hematol. Educ. Program 2010, 377-384 (2010).

[0006] Current ITP treatments work in a variable fraction of patients and prediction of efficacy is impossible. Moreover, ITP treatments (e.g., corticosteroids) address the destruction component of the disease, without addressing underlying cause. Most therapies (including splenectomy) for long-term chronic use have significant tolerability issues, generally associated with immunosuppression, and for some agents, even mortality. Few of the agents have been tested in controlled clinical trials and none have demonstrated a significant reduction in the bleeding associated with the low platelet counts. The vast majority of drugs are prescribed off-label (e.g., azathioprine, danazol, vinca alkaloids and rituximab), and have variable and often only transient efficacy in patients with chronic ITP. Neunert et al., Blood, 117(16):4190-4207 (2011). The day to day clinical management of patients with chronic ITP is therefore, a significant problem.

[0007] The crucial role of CD40-CD40L interactions in immune and inflammatory responses has made them a promising target for treatment of pathological immuno-inflammatory processes. Blockade of CD40-CD40L interactions by means of specific CD40L monoclonal antibodies (mAbs) successfully prevents allograft rejection in primates and treats autoimmune diseases and atherosclerosis in animal models. Montgomery et al., Transplantation, 74:1365-1369 (2002). In humans, two different anti-CD40L mAb clones have been used in clinical trials for treatment of different autoimmune diseases. Maribel et al., Mol. Immunol., 45:937-944 (2008). Monoclonal antibodies, however, can display unusually high incidence of thromboembolic (TE) complications, such as atherothrombotic central nervous system events, myocardial infarction, pulmonary embolism, and deep vein thrombosis. For example, the usefulness of the anti-CD40L mAb clone hu5c8 (anti-CD40L mAb, Biogen) is limited by an unusually high incidence of TE complications. TE by these antibodies is thought to result from the formation of higher-order immune complexes (IC) of the mAbs with membrane-bound CD40L on platelets, or sCD40L shed from platelets, that can ligate and thereby aggregate neighboring platelets via their FcgRIIa receptors, resulting in thrombi formation. The risk of thromboembolism has led to a halt in all ongoing clinical trials. Boumpas et al., Arthritis Rheum., 48:719-727 (2003).

[0008] Accordingly, it is an object of this invention to provide improved methods for treating subjects with ITP and other autoimmune disorders without the risk of thromboembolism.

SUMMARY

[0009] In certain aspects, the present invention relates to use of antibody polypeptides that specifically bind and inhibit human CD40L (also referred to as “anti-CD40L antibody polypeptides”) and that are useful in the treatment of diseases involving CD40L activation are disclosed. The antibody polypeptides advantageously do not cause platelet aggregation. Ultimately, targeting CD40L with the antibody polypeptide can provide opportunity to inhibit autoimmune processes leading to, for example, primary immune thrombocytopenia, solid organ transplant rejection, graft-related disease, Pemphigus vulgaris, systemic sclerosis, and myasthenia gravis and induce durable disease remission. The antibody polypeptide can thus provide long lasting therapeutic benefits for autoimmune disease patients. The antibody polypeptide thus offers a novel therapeutic modality, currently not available to patients. Given the lack of therapeutic options for autoimmune disease subjects who have failed all conventional therapies, the antibody polypeptide with its distinct mechanism of action and known safety profile demonstrates a favorable Risk/Benefit profile.

[0010] Unmet medical need for ITP in particular includes: 1) management of severe, chronic ITP in patients who are intolerant of (due to treatment-associated side effects) or inappropriate for (due to comorbidities) treatment with non-selective immunosuppressive agents or refractory to available treatment options (including splenectomy), where the life-threatening bleeding risk remains high and quality of life decreases; 2) more effective and safer therapies, potentially curative and/or inducing long lasting remission (focusing on reducing PLT destruction) and able to reduce or eliminate need for CS and replace or significantly defer splenectomy.

[0011] A pharmaceutical composition is provided comprising a therapeutically-effective amount of the anti-CD40L antibody polypeptide and a pharmaceutically acceptable carrier. The pharmaceutical composition can further comprise an immunosuppressive/immunomodulatory and/or anti-inflammatory agent.

[0012] In certain embodiments, a method of treating an immune disease in a patient in need of such treatment is provided. Such method comprises administering to the patient a therapeutically effective amount of the anti-CD40L antibody polypeptide described herein. The immune disease can be an autoimmune disease or a graft-related disease. The immune disease can be selected from the group consisting of selected from the group consisting of Addison's disease, allergies, ankylosing spondylitis, asthma, atherosclerosis, autoimmune diseases of the ear, autoimmune diseases of the eye, autoimmune hepatitis, autoimmune parotitis, colitis, coronary heart disease, Crohn's disease, diabetes, including Type 1 and/or Type 2 diabetes, epididymitis, glomerulonephritis, graft-related disease, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia, idiopathic thrombocytopenic purpura (also known as primary immune thrombocytopenia), inflammatory bowel disease, immune response to recombinant drug products (e.g., Factor VII in hemophiliacs), systemic lupus erythematosus, male infertility, multiple sclerosis, myasthenia gravis, pemphigus (such as Pemphigus vulgaris and Pemphigus foliaceus), psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma (systemic sclerosis), Sjogren's syndrome, spondyloarthropathies, thyroiditis, transplant rejection, and vasculitis. Autoimmune-mediated conditions include, but are not limited to, conditions in which the tissue affected is the primary target, and in some cases, the secondary target. Such conditions include, but are not limited to, AIDS, atopic allergy, bronchial asthma, eczema, leprosy, schizophrenia, inherited depression, transplantation of tissues and organs (such as solid organ transplant), chronic fatigue syndrome, Alzheimer's disease, Parkinson's disease, myocardial infarction, stroke, autism, epilepsy, Arthus' phenomenon, anaphylaxis, alcohol addiction, and drug addiction. Furthermore, the graft-related disease can comprise solid organ, tissue and/or cell transplant rejection. Alternatively, the graft-related disease is graft versus host disease (GVHD). The graft-related disease can further be an acute transplant rejection. Alternatively, the graft-related disease can be a chronic transplant rejection. In a specific embodiment, the autoimmune disease is idiopathic thrombocytopenic purpura (ITP).

[0013] Also provided is a method of treating an immune disease in a patient, the method comprising administering to the patient a therapeutically-effective amount of an anti-CD40L antibody polypeptide (e.g., BMS-986004), wherein at least one dose of the antibody polypeptide is administered at a dose from about 75 mg to about 1500 mg. The dose can be at least about 75 mg, at least about 225 mg, or at least about 675 mg. The dose can be about 75 mg, about 225 mg, about 675 mg, or about 1500 mg. Alternatively, the dose may range from about 200 mg to about 1200 mg, or from about 500 mg to about 1000 mg. Optionally, the method comprises at least one administration cycle (e.g., 2 weeks).

[0014] The antibody polypeptide can be formulated in a pharmaceutical composition for intravenous administration. The pharmaceutical composition can comprise a pharmaceutically acceptable carrier. For example, the pharmaceutical composition comprises 10 mM sodium phosphate, pH 6.5, 25 mM arginine HCl, and 250 mM sucrose.

[0015] The antibody polypeptide can be administered intravenously. The dose of antibody polypeptide may be from about 200 to about 1500 mg. At least 2 doses can be administered. Optionally, at least 7 doses can be administered. When multiple doses are administered, the doses may be the same or different. The dose can be administered once every 2 weeks.

[0016] The method can normalize platelet counts in the patient. Optionally, the patient's baseline peripheral blood platelet count can at least double after treatment. The patient's peripheral blood baseline platelet count can be greater than or equal to 50,000/mm.sup.3 or greater than or equal to 100,000/mm.sup.3 after treatment. The patient can demonstrate a peripheral blood platelet count increase of 20,000/mm.sup.3 after treatment. The patient can have a peripheral blood platelet count of less than 30,000/mm.sup.3 before treatment. The patient can have a peripheral blood platelet count of less than 100×10.sup.9 L or 50×10.sup.9 L before treatment. The patient can be splenectomized prior to treatment. Optionally, the patient was previously treated for ITP before treatment. Alternatively, the patient was not previously treated for ITP before treatment.

[0017] In certain embodiments, the methods of the present invention further comprise administering to patient an immunosuppressive/immunomodulatory and/or anti-inflammatory agent. The immunosuppressive/immunomodulatory and/or anti-inflammatory agent and the anti-CD40L antibody polypeptide may be administered sequentially or concurrently.

[0018] A kit for treating an immune disease in a patient is also provided, the kit comprising: (a) a dose of an anti-CD40L antibody polypeptide (e.g., BMS-986004); and (b) instructions for using the antibody polypeptide.

[0019] Also provided is an anti-CD40L antibody polypeptide (e.g., BMS-986004) for administration, wherein at least one dose of the anti-CD40L antibody polypeptide is administered at a dose of about 75 to about 1500 mg.

[0020] Also provided is a use of an anti-CD40L antibody polypeptide disclosed herein for the preparation of a medicament for the treatment of a patient, wherein the patient has or is at risk of having an immune disease. Further provided is a use of an anti-CD40L antibody polypeptide disclosed herein for preparation of a medicament for alleviating at least one symptom of an immune disease in a patient in need thereof. Further provided is an anti-CD40L antibody polypeptide (e.g., BMS-986004) for use in at least one administration cycle, wherein for each cycle one dose of the antibody polypeptide is administered at a dose of about 75 to about 1500 mg.

[0021] The anti-CD40L antibody polypeptides comprise a variable domain. Exemplary antibody polypeptides are in the form of a domain antibody (dAb) that contains a single variable domain. Alternatively, the dAbs can be bi-specific reagents that comprise a second variable domain that can bind another antigen, such as human serum albumin (HSA), for example.

[0022] In certain embodiments, the antibody polypeptide of the invention comprises a first variable domain that specifically binds human CD40L, wherein CD40L comprises the amino acid sequence of SEQ ID NO: 1, wherein the amino acid sequence of the first variable domain comprises: (a) a CDR1 region which differs from the CDR1 region of BMS2h-572-633 (SEQ ID NO: 2) by up to three amino acids, (b) a CDR2 region which differs from the CDR2 region of BMS2h-572-633 ((SEQ ID NO: 3) by up to three amino acids, (c) a CDR3 region which differs from the CDR3 region of BMS2h-572-633 (SEQ ID NO: 4) by up to three amino acids; and wherein the antibody polypeptide inhibits binding of CD40L to CD40 with an EC50 of 100 pM to 100 nM. Alternatively, the amino acid sequence of the first variable domain can differ from the amino acid sequence of BMS2h-572-633 (SEQ ID NO: 5) by up to and including 10 amino acids. Furthermore, the amino acid sequence of the first variable domain can differ from SEQ ID NO: 5 by up to and including 5 amino acids. The amino acid sequence of the first variable domain can also differ from SEQ ID NO: 5 by up to and including 2 amino acids. Alternatively, the first variable domain differs from SEQ ID NO: 5 by 1 amino acid. Alternatively, the variable domain of the antibody polypeptide comprises an amino acid sequence that is at least 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 5.

[0023] In certain specific embodiments, the variable domain of the antibody polypeptide comprises: (1) a CDR1 region having the amino acid sequence of SEQ ID NO: 2; (2) a CDR2 region having the amino acid sequence of SEQ ID NO: 3; and (1) a CDR3 region having the amino acid sequence of SEQ ID NO: 4. For example, the variable domain of the antibody polypeptide comprises the amino acid sequence of SEQ ID NO: 5 (BMS2h-572-633). Preferably, the antibody polypeptide is BMS-986004 and comprises the amino acid sequence of SEQ ID NO: 6.

[0024] Also provided is an antibody polypeptide comprising a first variable domain that specifically binds human CD40L, wherein the antibody polypeptide is a domain antibody (dAb). The antibody polypeptide can be a fusion polypeptide comprising the first variable domain and an Fc domain. Alternatively, the fusion polypeptide can comprise an IgG4 Fc domain. The fusion polypeptide also can comprise an IgG1 Fc domain. The fusion polypeptide can also comprise an IgG1 Fc domain. Alternatively, the fusion polypeptide can comprise a CT-Long domain. The fusion polypeptide can also comprise a CT-short domain. Alternatively, the fusion polypeptide can comprise a N297Q Long Fc domain. The fusion polypeptide can alternatively comprise a N297Q Short Fc domain.

[0025] Also provided is an antibody polypeptide comprising a first variable domain that specifically binds human CD40L, wherein the antibody polypeptide further comprises a second variable domain that specifically binds a second antigen, wherein the second antigen is an antigen other than human CD40L. The second antigen can be a cluster of differentiation (CD) molecule or a Major Histocompatibility Complex (MHC) Class II molecule. Alternatively, the second antigen can be serum albumin (SA).