Methods for treating diseases using anti-IL1RAP antibodies and antibody-drug conjugates

Abstract

Disclosed herein are interleukin-1 receptor accessory protein (IL1RAP) antibodies, compositions and methods of using the antibodies.

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Application No. 63 / 358,858, filed July 6, 2022, the entire contents of which are expressly incorporated herein by reference. [Background technology]

[0002] IL1RAP, also known as "interleukin-1 receptor accessory protein," "IL-1R accessory protein," "IL-1 receptor accessory protein," "interleukin-1 receptor 3," "IL-1RAcP," "C3orf13," "IL-1R3," "interleukin-1 receptor accessory protein β," "interleukin-1 receptor accessory protein," and "IL-1R-3" (Wesche, HJ Biol. Chem. 272:7727-7731, 1997), is an essential part of the interleukin-1 (IL-1) receptor complex that initiates signaling events that lead to the activation of interleukin-1-responsive genes. In addition to IL1 signaling, IL1RAP is important for mediating the actions of IL-33 via the ST2 / IL1RAP complex and IL-36 via the IL1Rrp2 / IL1RAP complex (Garlanda et al., Immunity. 2013 Dec 12;39(6):1003-18).

[0003] Two IL-1 receptors, IL-1R type I and IL-1R type II, have been identified. Both receptors can interact with both forms of IL-1, i.e., IL-1α and IL-1β. IL-1R1 mediates IL-1-induced cell activation. However, the IL-1 / IL-1R1 complex cannot signal by itself but is dependent on its association with IL1RAP (Dinarello, CA, Blood 30 1996, 87(6):2095-147) (see, e.g., WO2015 / 132602).

[0004] Alternative splicing of IL1RAP generates two transcripts encoding two distinct isoforms, one membrane-bound and one soluble. The ratio of the soluble form to the membrane-bound form increases during acute phase induction or stress. IL1RAP is expressed in leukemic stem cell candidates in most AML patients, but not in normal hematopoietic stem cells (Ågerstam, et al. PNAS USA (2015) vol. 112:34, 10786-10791).

[0005] Interleukin-1 Interleukin-1 (IL-1) is a potent proinflammatory cytokine that induces the synthesis of acute-phase and proinflammatory proteins during infection, tissue injury, or stress by forming a complex with the interleukin-1 receptor and accessory proteins at the cell membrane. IL-1 can be produced by various cell types, including mononuclear phagocytes, in response to infection and inflammation. The IL-1 family consists of seven agonists, including IL-1α and IL-1β, and three naturally occurring receptor antagonists, including IL-1 receptor antagonist (IL-1Ra) (Dinarello, CA, Blood 1996, 87(6):2095-147).

[0006] IL-1 can activate several types of cells, including leukocytes and endothelial cells. IL-1 induces and amplifies immune responses by promoting the production and expression of adhesion molecules, cytokines, chemokines, and inflammatory mediators such as prostaglandin E2 and nitric oxide (NO). As a result, local inflammation is amplified and persists. Furthermore, IL-1-induced production of inflammatory mediators causes fever, headache, hypotension, and weight loss. Furthermore, IL-1 is a hematopoietic growth factor and has been shown to reduce leukocyte and platelet nadirs in patients undergoing bone marrow transplantation. IL-1 has also been shown to promote angiogenesis by inducing the production of vascular endothelial growth factor, thereby promoting pannus formation and blood supply in rheumatoid joints. Finally, IL-1 has been shown to promote bone and cartilage degradation in rheumatoid diseases.

[0007] IL-1 has been implicated in a wide range of diseases and conditions, ranging from gout to cancer (for reviews, see Dinarello et al., 2012, Nature Reviews 11:633-652 and Dinarello, 2014, Mol. Med. 20(suppl. 1):S43-S58). Several therapies to block IL-1 activity are approved and in development. Targeting IL-1 began in 1993 with the introduction of anakinra (Kineret™, Amgen), a recombinant form of the naturally occurring IL-1 receptor antagonist (IL-1Ra) that blocks both IL-1α and IL-1β activity. Since then, this therapeutic agent has been used to demonstrate the role of IL-1 in many diseases. Neutralizing IL-1 with antibodies or soluble receptors has also proven effective, with currently approved soluble decoy receptors (Arcalyst™, Regeneron) and the anti-IL-1β neutralizing monoclonal antibody canakinumab (Ilaris™, Novartis). Other therapeutic approaches, including IL-1α neutralization, therapeutic vaccines targeting IL-1β, and chimeric IL-1Ra, are in clinical trials. Additionally, orally active small molecule inhibitors of IL-1 production, such as caspase 1 inhibitors, are being developed.

[0008] However, there remains a need in the art for therapeutic agents that can be used to treat inflammatory, autoimmune, and atopic diseases, including atopic dermatitis and asthma, as well as suppurative inflammatory diseases such as pyoderma suppurativa. Summary of the Invention

[0009] Provided herein are anti-IL1RAP antibodies, antigen-binding portions thereof, and antibody-drug conjugates (ADCs) (e.g., as described in U.S. Pat. No. 11,248,054, which is incorporated by reference in its entirety for all purposes) for use in treating various diseases, disorders, and conditions, including, for example, inflammatory or autoimmune diseases such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, etc.

[0010] In another aspect of the present invention, the present disclosure provides a method for treating an inflammatory or autoimmune disease, disorder, or condition, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding portion thereof described herein.

[0011] In some embodiments, the inflammatory or autoimmune disease is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, the inflammatory or autoimmune disease is an atopic disease. In some embodiments, the inflammatory or autoimmune disease is characterized by neutrophil or eosinophil dysfunction. In some embodiments, the treatment reduces the level of TARK / CCL17 at atopic sites or in serum. In some embodiments, the inflammatory or autoimmune disease is atopic dermatitis.

[0012] In some embodiments, the present disclosure provides a method for inhibiting or reducing an inflammatory or autoimmune response in a subject having inflammation or autoimmunity, the method comprising administering to the subject having inflammation or autoimmunity an effective amount of an antibody or antigen-binding portion thereof described herein, such that the inflammation or autoimmunity is inhibited or reduced.

[0013] In some embodiments, the method of treating an inflammatory or autoimmune disease, disorder or condition, or inhibiting or reducing an inflammatory or autoimmune response, comprises identifying a mammal in need of such treatment according to abnormal serum levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, and IL-36γ, or CXCL1, CXCL8, TARC, LCN2, or combinations thereof.

[0014] In some embodiments, the antibody or antigen-binding portion thereof is characterized by its inhibition of IL-8 release by intestinal epithelial cells, intestinal myofibroblasts, or skin fibroblasts stimulated with IL-1α, IL-1β, IL-36α, IL-36β, and IL-36γ.

[0015] In some embodiments, the antibody or antigen-binding portion thereof is administered in combination with at least one other agent or therapy. In some embodiments, the at least one other agent or therapy is a therapeutic antibody, a small molecule drug, an siRNA, an mRNA, or any other modality known to those of skill in the art. In some embodiments, the antibody or antigen-binding portion thereof is administered before, simultaneously with, or after the at least one other agent or therapy. In some embodiments, the at least one other agent or therapy inhibits a TH2-related immune response, a TH1-related immune response, pruritus, IL-4 signaling, IL-13 signaling, IL-22 signaling, IL-33 signaling, IL-17 signaling, IL-36 signaling, IL-18 signaling, IL-23 signaling, OX40 signaling, IL-5 signaling, T cell migration, IRAK4 signaling, complement signaling, PDE4 signaling, or a combination thereof.

[0016] Therefore, this book contains the following: 1. A method of treating an inflammatory or autoimmune condition in a mammal in need thereof, comprising administering a therapeutically effective amount of an antibody against IL1RAP. 2. The method according to item 1, wherein the inflammatory or autoimmune condition is an atopic condition, and the antibody reduces the level of TARC / CCL17, PARC, periostin, IL-22, eotaxin-1, eotaxin-3, or a combination thereof in the atopic site or serum. 3. The method of claim 1, wherein the inflammatory or autoimmune condition is characterized by or caused by an increased number of neutrophil or eosinophil cells, neutrophil or eosinophil dysfunction, or increased TARC / CCL17 levels. 4. The method according to item 3, wherein the inflammatory or autoimmune condition is characterized or caused by an increased number of neutrophil cells or neutrophil dysfunction. 5. The method according to item 4, wherein the inflammatory or autoimmune condition characterized by or caused by an increased number of neutrophil cells or neutrophil dysfunction is selected from the group consisting of hidradenitis suppurativa, generalized pustular psoriasis (GPP), COPD, idiopathic fibrosis, neutrophilic asthma, neutrophilic dermatosis, pyoderma gangrenosum-Schnitzler syndrome, Behcet's disease, Sweet's syndrome, rheumatoid arthritis, systemic lupus erythematosus (SLE), inflammatory bowel disease (Crohn's disease, ulcerative colitis), psoriasis, vasculitis, Alzheimer's disease, COVID-19, and gout. 6. The method according to item 3, wherein the inflammatory or autoimmune condition is further characterized by an increased number of eosinophil cells or eosinophil dysfunction. 7. The method according to item 6, wherein the inflammatory or autoimmune condition characterized by or caused by an increased number of eosinophil cells or eosinophil dysfunction is selected from the group consisting of atopic dermatitis (eczema), allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, and eosinophilic myocarditis. 8. The method according to item 3, wherein the inflammatory or autoimmune condition is further characterized by increased TARC / CCL17 levels. 9. The method according to item 8, wherein the inflammatory or autoimmune condition characterized by an increased TARC / CCL17 level is selected from the group consisting of atopic dermatitis, COPD, bronchial asthma, allergic rhinitis, eosinophilic pneumonia, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, mastocytosis, and eosinophil-associated diseases. 10. The method according to item 1, wherein the inflammatory or autoimmune condition is sepsis, acute respiratory distress syndrome, COVID-19, myocardial infarction, cystic fibrosis, irritable bowel disease, ulcerative colitis, Crohn's disease, atopic dermatitis, psoriasis, multiple sclerosis, asthma, neutrophilic asthma, Alzheimer's disease, stroke, diabetic kidney disease, diabetic retinopathy, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, non-alcoholic fatty liver disease, or rheumatoid arthritis. inflammatory bowel disease, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, systemic lupus erythematosus (SLE), vasculitis, gout, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorders, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, and mastocytosis. 11. The method according to item 1, further comprising identifying a mammal in need thereof according to abnormal serum levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof. 12. The method according to item 1, further comprising identifying a mammal in need thereof according to abnormal serum levels of CXCL1, CXCL8, LCN-2, TARC, or a combination thereof. 13. The method according to item 11, wherein the serum level of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof is elevated. 14. The method according to item 12, wherein the serum levels of CXCL1, CXCL8, LCN-2, TARC, or a combination thereof are elevated. 15. The method of claim 1, wherein the antibody inhibits IL-8 release by intestinal epithelial cells, intestinal myofibroblasts, or skin fibroblasts stimulated with IL-1α, IL-1β, IL-36α, IL-36β, and / or IL-36γ, or a combination thereof. 16. The method according to item 2, wherein the atopic condition is selected from the group consisting of atopic dermatitis, asthma, COPD, allergic rhinitis, allergic conjunctivitis, food allergy, drug allergy, and angioedema. 17. The method according to item 2, wherein the atopic condition is atopic dermatitis, and the antibody reduces the level of TARC / CCL17, PARC, periostin, IL-22, eotaxin-1, eotaxin-3, or a combination thereof in the skin or serum. 18. The method according to item 2, wherein the atopic condition is asthma. 19. The method according to item 2, wherein the atopic condition is COPD. 20. The method of claim 1, wherein the antibody is administered in combination with at least one other therapeutic agent. 21. The method according to item 20, wherein the at least one other therapeutic agent is a therapeutic antibody, a corticosteroid, a small molecule, an siRNA, an mRNA, or a combination thereof. 22. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-4 signaling. 23. The method according to item 22, wherein the at least one other therapeutic agent is an IL-4Rα inhibitor or antagonist, a pan-JAK inhibitor or antagonist, or a combination thereof. 24. The method according to item 22, wherein the at least one other therapeutic agent is dupilumab, CBP-201, AK120, celdulatinib, CEE321, yaktinib, delgocitinib, filgotinib, tofacitinib, dexamethasone, triamcinolone, prednisone, or a combination thereof. 25. The method according to item 22, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, polyarticular juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 26. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-13 signaling. 27. The method according to item 26, wherein the at least one other therapeutic agent is an IL-13 inhibitor or antagonist, a JAK1 inhibitor or antagonist, a TYK2 inhibitor or antagonist, or a combination thereof. 28. The method according to item 26, wherein the at least one other therapeutic agent is upadacitinib, abrocitinib, tralokinumab, lebrikizumab, ebrasakimab, baricitinib, ruxolitinib, filgotinib, PF-06651600, dexamethasone, triamcinolone, prednisone, or a combination thereof. 29. The method according to item 26, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, systemic lupus erythematosus, vitiligo, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD or asthma. 30. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-22 signaling. 31. The method of claim 30, wherein the at least one other therapeutic agent is an IL-22 inhibitor or antagonist, an IL-22R1 inhibitor or antagonist, a JAK1 inhibitor or antagonist, a JAK2 inhibitor or antagonist, a TYK2 inhibitor or antagonist, or a combination thereof. 32. The method according to item 30, wherein the at least one other therapeutic agent is fezakinumab, LEO138559, brepositinib, ATI-1777, deuclavacitinib, TAK-279, upadacitonib, and abrocitinib, or a combination thereof. 33. The method according to item 30, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, systemic lupus erythematosus, vitiligo, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 34. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-33 signaling. 35. The method according to item 34, wherein the at least one other therapeutic agent is an IL-33 or IL-33R inhibitor or antagonist. 36. The method according to item 34, wherein the at least one other therapeutic agent is etokimab, itepekimab, astegolimab, PF-06817024, tozorakimab, CNTO7160, or a combination thereof. 37. The method according to item 34, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, chronic obstructive pulmonary disease (COPD), asthma, allergic contact dermatitis, irritant contact dermatitis, rosacea, plaque psoriasis, pustular psoriasis, mastocytosis, systemic lupus erythematosus, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, Behcet's disease, or vitiligo. 38. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-17 signaling. 39. The method according to item 38, wherein the at least one other therapeutic agent is an IL-17A, IL-17C, IL-17F, IL17A / F, or IL-17RA inhibitor or antagonist, or a combination thereof. 40. The method according to item 38, wherein the at least one other therapeutic agent is secukinumab, ixekizumab, brodalumab, bimekizumab, izokibep, sonelokimab, or a combination thereof. 41. The method according to item 38, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, tendonitis-associated arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COPD, or asthma. 42. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-36 signaling. 43. The method according to item 42, wherein the at least one other therapeutic agent is an IL-36 inhibitor or antagonist, or an IL-36R inhibitor or antagonist. 44. The method according to item 42, wherein the at least one other therapeutic agent is spesolimab, imsidolimab, REGN6490, or a combination thereof. 45. The method of claim 42, wherein the inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, neutrophilic asthma, neutrophilic pneumonia, COVID-19, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, nonalcoholic fatty liver disease, neutrophilic dermatosis, The method is hidradenitis suppurativa, generalized pustular psoriasis, palmoplantar pustular psoriasis, IL-36 receptor antagonist (DITRA) deficiency, plaque psoriasis, CARD14-mediated psoriasis, acute generalized exanthematous pustulosis, pyoderma gangrenosum, Sweet's syndrome, systemic lupus erythematosus, systemic sclerosis, autoimmune bullous disease, acne, allergic contact dermatitis, or folliculitis and eosinophilic pustular folliculitis. 46. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-18 signaling. 47. The method according to item 46, wherein the at least one other therapeutic agent is an IL-18 inhibitor or antagonist. 48. The method according to item 46, wherein the at least one other therapeutic agent is tadequinig alfa. 49. The method according to item 46, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, adult-onset Still's disease, cutaneous lupus erythematosus, chronic spontaneous urticaria, contact dermatitis, alopecia areata, cutaneous drug eruption, graft-versus-host disease, cryopyrin-associated periodic syndrome, granulomatosis with polyangiitis, systemic sclerosis, hidradenitis suppurativa, septic arthritis, pyoderma gangrenosum and acne (PAPA), familial Mediterranean fever, rosacea, synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO), bullous pemphigoid, pemphigus vulgaris, Behcet's disease, or Schnitzler's syndrome. 50. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-23 signaling. 51. The method of item 50, wherein the at least one other therapeutic agent is an IL-23 inhibitor or antagonist, an IL-12 / 23p40 subunit inhibitor or antagonist, an IL-23p19 subunit inhibitor or antagonist, or a combination thereof. 52. The method according to item 50, wherein the at least one other therapeutic agent is risankizumab, ustekinumab, guselkumab, tildrakizumab, mirikizumab, brazikumab, or a combination thereof. 53. The method according to item 50, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 54. The method according to item 20, wherein the at least one other therapeutic agent inhibits OX40 signaling. 55. The method according to item 54, wherein the at least one other therapeutic agent is an OX40 or OX40L inhibitor or antagonist. 56. The method according to item 54, wherein the at least one other therapeutic agent is locatinlimab, GBR830, amritelimab, or a combination thereof. 57. The method according to item 54, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 58. The method according to item 20, wherein the at least one other therapeutic agent inhibits IL-5 signaling. 59. The method according to item 58, wherein the at least one other therapeutic agent is an IL-5Rα inhibitor or antagonist, a JAK1 inhibitor or antagonist, or a combination thereof. 60. The method according to item 58, wherein the at least one other therapeutic agent is benralizumab, upadacitinib, abrocitinib, SHR0302, filgotinib, PF-06651600, dexamethasone, triamcinolone, prednisone, or a combination thereof. 61. The method according to item 58, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 62. The method according to item 20, wherein the at least one other therapeutic agent inhibits T cell migration. 63. The method according to item 62, wherein the at least one other therapeutic agent is an S1PR1 inhibitor or antagonist, an S1PR4 inhibitor or antagonist, an S1PR5 inhibitor or antagonist, a CCR4 inhibitor or antagonist, or a combination thereof. 64. The method according to item 62, wherein the at least one other therapeutic agent is etrasimod, ozanimod, SCD-044, LC51-0255, BMS-986166, RPT193, or a combination thereof. 65. The method according to item 62, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 66. The method according to item 20, wherein the at least one other therapeutic agent inhibits pruritus. 67. The method according to item 66, wherein the at least one other therapeutic agent is an IL-1α inhibitor or antagonist, an OSMRβ inhibitor or antagonist, an NK1R inhibitor or antagonist, a P2X3 inhibitor or antagonist, an IL-31 inhibitor or antagonist, or a combination thereof. 68. The method according to item 66, wherein the at least one other therapeutic agent is bermekimab, bicsarelimab, serlopitant, tradipitant, BLU-5937, nemolizumab, or a combination thereof. 69. The method according to item 66, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma. 70. The method according to item 20, wherein the at least one other therapeutic agent inhibits a TH1-associated immune response. 71. The method according to item 70, wherein the at least one other therapeutic agent is an IL-1α inhibitor or antagonist, an IL-1β inhibitor or antagonist, an IL-1R1 inhibitor or antagonist, an IL-36R inhibitor or antagonist, a TNFα inhibitor or antagonist, or a combination thereof. 72. The method according to item 70, wherein the at least one other therapeutic agent is bermekimab, anakinra, canakinumab, gevokizumab, rilonacept, MEDI8968, spesolimab, imsidolimab, REGN6490, adalimumab, infliximab, etanercept, certolizumab, golimumab, or a combination thereof. 73. The method according to item 70, wherein the inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, and familial idiopathic rheumatoid arthritis. Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, acne (PAPA), cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS) also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar psoriasis The method is pustular psoriasis (PPP), dermatomyositis, panniculitis, Erdheim-Chester syndrome, adenosine deaminase deficiency (DADA2), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), neutrophilic dermatosis, or a solitary autoinflammatory skin disease. 74. The method according to item 20, wherein the at least one other therapeutic agent inhibits a TH2-associated immune response. 75. The method of claim 74, wherein the at least one other therapeutic agent is an IL-4 inhibitor or antagonist, a type I IL-4 receptor inhibitor or antagonist, a type II IL-4 receptor inhibitor or antagonist, an IL-13 inhibitor or antagonist, a type I IL-13 receptor inhibitor or antagonist, a type II IL-13 receptor inhibitor or antagonist, and an IL-5 inhibitor or antagonist, a type I IL-5 receptor inhibitor or antagonist, a type II IL-5 receptor inhibitor or antagonist, an IL-9 inhibitor or antagonist, a type I IL-9 receptor inhibitor or antagonist, an IL-10 inhibitor or antagonist, a homodimeric IL-10 receptor inhibitor or antagonist, a heterodimeric IL-10 receptor inhibitor or antagonist, or a combination thereof. 76. The method according to item 74, wherein the at least one other therapeutic agent is dupilumab, omalizumab, mepolizumab, benralizumab, tralokinumab, lebrikizumab, or a combination thereof. 77. The method according to item 70, wherein the inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, familial Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, and acne (PA). PA), cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis, panniculitis, Erdheim-Chester syndrome, adenosine deaminase deficiency (ADA2) ), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), neutrophilic skin The method is characterized by the following: pyrin-associated autoinflammation with inflammatory bowel disease (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), neutrophilic dermatosis or isolated autoinflammatory skin disease, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, or urticaria. 78. The method according to item 20, wherein the at least one other therapeutic agent inhibits a TH17-associated immune response. 79. The method according to item 78, wherein the at least one other therapeutic agent is an IL-17 inhibitor or antagonist, a type I IL-17 receptor inhibitor or antagonist, a type II IL-17 receptor inhibitor or antagonist, an IL-23 inhibitor or antagonist, an IL-23 receptor inhibitor or antagonist, or a combination thereof. 80. The method according to item 78, wherein the at least one other therapeutic agent is secukinumab, ixekizumab, brodalumab, bimekizumab, izokibep, sonelokimab, risankizumab, ustekinumab, guselkumab, tildrakizumab, mirikizumab, brazikumab, or a combination thereof. 81. The method according to item 78, wherein the inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticaria vasculitis, familial Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, acne vulgaris (PAPA), and psoriasis-associated inflammatory diseases. Cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis, panniculitis, Erdheim-Chester syndrome, adenosine deaminase 2 (DADA2) deficiency, Majid syndrome group, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammatory disease with neutrophilic dermatosis and the method is characterized by the following: allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, or isolated autoinflammatory skin disorder. 82. The method according to item 20, wherein the at least one other therapeutic agent inhibits IRAK4 signaling. 83. The method according to item 82, wherein the at least one other therapeutic agent is an IRAK4 inhibitor, degrader, or antagonist. 84. The method according to item 82, wherein the at least one other therapeutic agent is PF-06650833, CA-4948, KT-474, or a combination thereof. 85. The method according to item 82, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COPD, or asthma. 86. The method according to item 20, wherein the at least one other therapeutic agent inhibits complement signaling. 87. The method of item 86, wherein the at least one other therapeutic agent is a C5a inhibitor or antagonist, a C5aR inhibitor or antagonist, a TSLP inhibitor or antagonist, a CD80 / CD86 inhibitor or antagonist, an IL-6 inhibitor or antagonist, a CD20 inhibitor or antagonist, an integrin α4 inhibitor or antagonist, an AhR agonist, or a combination thereof. 88. The method according to item 86, wherein the at least one other therapeutic agent is vilobelimab, FX002, INF904, tezepelumab, abatacept, tocilizumab, sarilumab, rituximab, vedolizumab, tapinarofibrate, tadekinig alfa, or a combination thereof. 89. The method of claim 86, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COVID-19, COPD, or asthma. 90. The method according to item 20, wherein the at least one other therapeutic agent inhibits PDE4 signaling. 91. The method according to item 90, wherein the at least one other therapeutic agent is a PDE4 inhibitor or antagonist. 92. The method according to item 90, wherein the at least one other therapeutic agent is apremilast, crisaborole, difamilast, roflumilast, or a combination thereof. 93. The method according to item 90, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, pustular purpura, COPD, or asthma. 94. The method according to item 20, wherein the at least one other therapeutic agent is administered before, simultaneously with, or after the at least one other therapeutic agent. 95. The method according to items 1, 17 and 20, wherein the antibody is administered subcutaneously or intravenously at a dose selected from 1 to 80 mg / kg, 1 to 60 mg / kg, 1 to 50 mg / kg, 1 to 40 mg / kg, 1 to 30 mg / kg, 1 to 25 mg / kg, or 1 to 20 mg / kg. 98. The method according to items 1, 17 and 20, wherein the antibody is administered as a subcutaneous injection or an intravenous bolus less frequently than once a week, twice a week, more frequently than twice a week, or by continuous infusion. 101. The method according to item 1, 17 or 20, wherein the antibody is a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 67, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 66, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 70, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 69; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 15, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 51, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 50, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 49, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 55, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 54, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 53; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 59, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 58, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 57, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 63, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 61; or A method comprising BFB759 (i.e., 37E10_15B5) corresponding to a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 176, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 175, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 174, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 179, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 178, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 79. 102. The method according to item 1, 17 or 20, wherein the antibody is a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 68; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 82; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 48 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 52; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 60; or A method comprising BFB759 (i.e., 37E10_15B5) corresponding to a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 173 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177. [Brief explanation of the drawings]

[0017] [Figure 1] 1 shows the surface expression of IL1RAP in human acute myeloid leukemia cell lines EOL1, Monomac6, OCI / AML1, and KG-1, and the T-cell leukemia cell line Karpas 299, as determined by flow cytometry analysis. [Figure 2] 1 shows the specific binding of IL1RAP antibody 44E5_15C5 to the IL1RAP-positive cell lines EOL1 and Karpas 299. The IL1RAP-negative cell line DMS79 does not show binding by IL1RAP antibody 44E5_15C5. [Figure 3] The antibody competing bin layout is shown. [Figure 4A] Figure 1 shows binding of anti-IL1RAP antibodies to IL1RAP orthologs. Anti-IL1RAP antibodies were assessed for cell surface binding to 293 cells expressing human IL1RAP by flow cytometry. 21H5 was a murine antibody against human IL1RAP. CB1gG1 (anti-hen egg lysozyme antibody, CrownBio™) was used as a negative control. [Figure 4B] Figure 1 shows binding of anti-IL1RAP antibodies to IL1RAP orthologs. Anti-IL1RAP antibodies were assessed for cell surface binding to 293 cells expressing macaca fascicularis by flow cytometry. 21H5 was a mouse antibody against human IL1RAP. CB1gG1 (anti-hen egg lysozyme antibody, CrownBio™) was used as a negative control. [Figure 4C] Figure 1 shows binding of anti-IL1RAP antibodies to IL1RAP orthologs. Anti-IL1RAP antibodies were assessed for cell surface binding to 293 cells expressing rat IL1RAP by flow cytometry. 21H5 was a mouse antibody against human IL1RAP. CB1gG1 (anti-hen egg lysozyme antibody, CrownBio™) was used as a negative control. [Figure 4D]Figure 1 shows binding of anti-IL1RAP antibodies to IL1RAP orthologs. Anti-IL1RAP antibodies were assessed for cell surface binding to 293 cells expressing mouse IL1RAP by flow cytometry. 21H5 was a mouse antibody against human IL1RAP. CB1gG1 (anti-hen egg lysozyme antibody, CrownBio™) was used as a negative control. [Figure 5] Internalization of IL1RAP antibody 44E5_15C5 into EOL1 cells. Live EOL1 cells were incubated with IL1RAP antibody 44E5_15C5 for 0.5 hours at 37°C. After cytospin, the cells were fixed, permeabilized, and co-stained with LAMP1 antibody. [Figure 6A] 1 shows blockade of IL1α / β signaling by anti-IL1RAP antibodies. IL1RAP antibodies 37E10_15B5, 44E5_15C5, 16H2_17D2, and 36A10_21B6 showed potent inhibition of IL1R1 signaling in a dose-dependent manner. [Figure 6B] Figure 1 shows blockade of IL1α / β signaling by anti-IL1RAP antibodies. Antibodies 37E10_15B5 and 44E5_15C5 inhibit IL-1β and IL-1α signaling with subnanomolar EC50. [Figure 6C] Figure 1 shows blockade of IL1α / β signaling by anti-IL1RAP antibodies. Antibodies 37E10_15B5 and 44E5_15C5 inhibit IL-1β and IL-1α signaling with subnanomolar EC50. [Figure 7]Blockade of IL-33 signaling by anti-IL1RAP antibody is shown. HEK-Blue IL-33 cells (Invivogen, CA) were harvested and plated in technical duplicates at a density of 50,000 cells per well in 96-well plates. Antibodies or corresponding human IgG1 control antibodies were added to wells at concentrations of 1 μg / ml and 10 μg / ml (A) or serial dilutions starting at 10 μg / ml (B). After 30 minutes of incubation with antibody, IL-33 was added to a final concentration of 0.5 ng / ml, and plates were incubated overnight. After 24 hours, substrate was added to the supernatant, and samples were analyzed for absorbance at 620 nm. CBIgG1 (anti-hen egg lysozyme antibody, CrownBio™) was used as a negative control. [Figure 8] 1 shows the experimental design for testing the efficacy of anti-IL1RAP antibodies in a mouse model of atopic dermatitis. [Figure 9] 1 shows the reduction of pruritus by the anti-IL1RAP antibody of the present invention in a mouse model of atopic dermatitis. [Figure 10] Figure 1 shows the results of the present invention: (A) suppression of weight gain by the anti-IL1RAP antibody of the present invention in a mouse model of atopic dermatitis; (B) prevention or reduction of skin thickening by the anti-IL1RAP antibody of the present invention in a mouse model of atopic dermatitis; and (C) prevention or reduction of splenomegaly by the anti-IL1RAP antibody of the present invention in a mouse model of atopic dermatitis. [Figure 11] 1 shows that the anti-IL1RAP antibody of the present invention reduced the levels of inflammatory / atopic mediators eotaxin, lipocalin-2, TARC / CCL17, TSLP, and IL-6 in the skin of atopic dermatitis model mice. [Figure 12] 1 shows that the anti-IL1RAP antibody of the present invention reduced the levels of neutrophils, macrophages, and eosinophils (ie, inflammatory cells) in the skin of atopic dermatitis model mice. [Figure 13]FB759 inhibits HDM-induced TARC levels in PBMCs, and dupilumab does not or minimally inhibits TARC in the same assay. For the data shown in Figure 13, hIgG4 (human IgG4 isotype Ab), BFB759, and dupilumab were used at 30 μg / ml, and HDM was used at 10 μg / ml. Data shown are mean ± SD. [Figure 14] Figure 14 shows a summary of the inhibition of HDM-induced TARC levels in PBMCs from different donors by BFB759. For the data shown in Figure 14, HDM was used at 10 μg / ml, BFB759 at 1-30 μg / ml, and PBMCs were obtained from 12 different donors. Data shown are mean ± SD. [Figure 15] These results demonstrate that BFB759 and dupilumab can inhibit TARC levels in PBMCs treated with HDM and IL-4. For the data shown in Figure 15, HDM was used at 10 μg / ml, IL-4 at 750 pg / ml, and antibodies were used at three different concentrations as indicated on the graph. Data shown are mean ± SD. [Figure 16] These results demonstrate that BFB759 and tralokinumab can inhibit TARC levels in PBMCs treated with HDM and IL-13. For the data shown in Figure 16, HDM was used at 10 μg / ml, IL-13 at 0.5 nM, and BFB759 at 1 μg / ml, with tralokinumab gradually titrated from 10 μg / ml to 0.16 μg / ml (four-fold dilutions, four series). Data shown are mean ± SD. [Figure 17A] These results demonstrate that the combination of BFB759 with a JAK inhibitor or dexamethasone can inhibit TARC in HDM-treated PBMCs. For the data shown in Figure 17A, HDM was used at 10 μg / ml, BFB759 was used at 1 μg / ml, and the JAKi and dexamethasone were tested at three different concentrations (0.1 nM, 10 nM, and 1 μM) as indicated on the graph. Data shown are mean ± SD. [Figure 17B]These results demonstrate that the combination of BFB759 with a JAK inhibitor or dexamethasone can inhibit TARC in HDM-treated PBMCs. For the data shown in Figure 17B, HDM was used at 10 μg / ml, BFB759 was used at 1 μg / ml, and the JAKi and dexamethasone were tested at three different concentrations (0.1 nM, 10 nM, and 1 μM) as indicated on the graph. Data shown are mean ± SD. [Figure 18A] The combination of BFB759 and upadacitinib inhibits TARC in PBMCs treated with HDM. For the data shown in Figure 18A, HDM was used at 10 μg / ml, BFB759 and hIgG4 (human IgG4 isotype Ab) were used at 1 μg / ml, and upadacitinib was tested in a 5-fold titration series from 1 μM to 0.01 nM. Data shown are mean ± SD. [Figure 18B] Figure 18B shows that the combination of BFB759 and upadacitinib inhibits TARC in PBMCs treated with HDM. For the data shown in Figure 18B, HDM was used at 10 μg / ml, BFB759 and hIgG4 (human IgG4 isotype Ab) were used at 1 μg / ml, and upadacitinib was tested in a 5-fold decrement series from 1 μM to 0.01 nM. Data shown are mean ± SD. [Figure 19A] Figure 19A shows that the combination of BFB759 and adalimumab inhibits IL-6 secretion in human whole blood cultures stimulated with HKCA. In the data shown in Figure 19A, IL-6 levels from whole blood cultures are measured by ELISA. [Figure 19B] The combination of BFB759 and adalimumab inhibits IL-6 secretion in human whole blood cultures stimulated with HKCA. In the data shown in Figure 19B, the percentage of IL-6 measured in cultures treated as indicated is compared to the level detected with heat-killed Candida albicans (HKCA) and without any intervention. [Figure 20A]1 shows that the combination of BFB759 and secukinumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with a combination of cytokines. [Figure 20B] 1 shows that the combination of BFB759 and secukinumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with a combination of cytokines. [Figure 21A] 1 shows that the combination of BFB759 and adalimumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with a combination of cytokines. [Figure 21B] 1 shows that the combination of BFB759 and adalimumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with a combination of cytokines. DETAILED DESCRIPTION OF THE INVENTION

[0018] Various aspects of the present disclosure relate to anti-IL1RAP antibodies, such as BFB759, and antibody fragments thereof, as described herein for binding to and inhibiting human IL1RAP on IL1RAP-expressing cells, inhibiting IL-1, e.g., IL-1β and / or IL-1α, IL-33, IL-36 (IL-36α, IL-36β, IL-36γ) signaling in vivo, and / or treating inflammatory or autoimmune diseases, disorders, or conditions, such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, and pharmaceutical compositions thereof.

[0019] In a specific embodiment, an anti-IL1RAP antibody of the invention, e.g., BFB759 of the invention, or an antigen-binding portion thereof, is administered in combination with one or more inhibitors or antagonists of a TH2-associated immune response, a TH1-associated immune response, a TH17-associated immune response, pruritus, IL-4 signaling, IL-13 signaling, IL-22 signaling, IL-33 signaling, IL-17 signaling, IL-36 signaling, IL-18 signaling, IL-23 signaling, OX40 signaling, IL-5 signaling, T cell migration, IRAK4 signaling, complement signaling, PDE4 signaling, or a combination thereof, for the treatment of an inflammatory or autoimmune disease or disorder, such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, etc.

[0020] I. Definition In order that the present invention may be more readily understood, certain terms are first defined. Additionally, it should be noted that whenever a value or range of values for a parameter is given, it is intended that values and ranges intermediate to the given values are also part of the present invention.

[0021] The terms "interleukin-1 receptor accessory protein antibody" or "anti-IL1RAP antibody," used interchangeably herein, refer to an antibody that specifically binds to IL1RAP, e.g., human IL1RAP. An antibody that "binds" an antigen of interest, i.e., IL1RAP, is one that is capable of binding to that antigen with sufficient affinity so that the antibody is useful for targeting cells expressing the antigen. In a preferred embodiment, the antibody specifically binds to human IL1RAP (hIL1RAP). Examples of anti-IL1RAP antibodies are disclosed in the Examples below and specified in Table 5. Unless otherwise specified, the term "anti-IL1RAP antibody" is intended to refer to an antibody that binds to wild-type IL1RAP, a variant or isoform of IL1RAP.

[0022] As described below, in a specific embodiment, a human anti-IL1RAP antibody for use in the methods of the invention herein is designated BFB759 (i.e., 37E10_15B5), which corresponds to a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 67, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 66, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 70, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 69. In another specific embodiment, the human anti-IL1RAP antibody BFB759 (i.e., 37E10_15B5) corresponds to a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 68.

[0023] Several different isoforms of IL1RAP have been identified. An exemplary amino acid sequence of wild-type human IL1RAP, comprising 570 amino acids, is provided below as SEQ ID NO: 260. The extracellular domain (ECD) of IL1RAP comprises amino acids 21 to 367 of SEQ ID NO: 260. [ka]

[0024] As used herein, the term "specific binding" or "specific binding" in reference to the interaction of an IL1RAP antibody, or antigen-binding portion thereof, with a second chemical species means that the interaction is dependent on the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species. For example, an antibody recognizes and binds to a particular protein structure, rather than proteins in general. If an antibody is specific for epitope "A," then in a reaction involving labeled "A" and the antibody, the presence of a molecule containing epitope "A" (or free, unlabeled "A") will reduce the amount of labeled "A" that binds to the antibody.

[0025]

[0023] In one embodiment, the phrase "specifically binds to hIL1RAP" or "specific binding to hIL1RAP" as used herein refers to a dissociation constant (K) of about 2,000 nM or less, about 1,000 nM or less, about 500 nM, about 200 nM or less, about 100 nM or less, about 75 nM or less, about 25 nM or less, about 21 nM or less, about 12 nM or less, about 11 nM or less, about 10 nM or less, about 9 nM or less, about 8 nM or less, about 7 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, about 1 nM or less, about 0.5 nM or less, about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM or less, or about 0.001 nM or less. D ) to IL1RAP (human or cynomolgus IL1RAP). In another embodiment, the phrase "specifically binds to hIL1RAP" or "specific binding to hIL1RAP" as used herein refers to a dissociation constant (K) of between about 1 pM (0.001 nM) and 2,000 nM, between about 500 pM (0.5 nM) and 1,000 nM, between about 500 pM (0.5 nM) and 500 nM, between about 1 nM and 200 nM, between about 1 nM and 100 nM, between about 1 nM and 50 nM, between about 1 nM and 20 nM, or between about 1 nM and 5 nM. D ) refers to the ability of an anti-IL1RAP antibody to interact with hIL1RAP at K D is determined by surface plasmon resonance or biolayer interferometry, or other methods known in the art. Biolayer interferometry refers to an optical phenomenon that allows for the analysis of real-time biospecific interactions by measuring the interference pattern of reflected white light, for example, using the Octet™ system (ForteBio, Pall Corp. Fremont, CA). Details of the Octet™ system are described in Li, B et al. (2011) J. Pharm. Biomed. Anal. 54(2):286-294 and Abdiche, YN, et al. (2009) Anal. Biochem. 386(2):172-180, the contents of which are incorporated herein by reference.

[0026] The term "antibody" refers broadly to an immunoglobulin (Ig) molecule, generally composed of four polypeptide chains: two heavy (H) chains and two light (L) chains, or any functional fragment, mutant, variant, or derivative thereof that retains the essential target binding function of an Ig molecule. Such mutant, modified, or derivative antibody formats are known in the art, non-limiting examples of which are described below.

[0027] The terms TH1 and TH-1, TH2 and TH-2, and TH17 and TH-17 are used interchangeably herein. The terms "mediated" and "associated" are used interchangeably herein.

[0028] In full-length antibodies, each heavy chain is composed of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains: CH1, CH2, and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is composed of one domain, CL. The VH and VL regions can be further divided into regions of hypervariability called complementarity-determining regions (CDRs), which are interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) and class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) or subclass.

[0029] As used herein, the term "antigen-binding portion" (or simply "antibody portion") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., hIL1RAP). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Embodiments of such antibodies may also be bispecific, dual specific, or multispecific, i.e., specifically bind to two or more different antigens. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) a Fab fragment: a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment: a bivalent fragment consisting of two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single antibody arm; (v) a dAb fragment comprising a single variable domain (Ward et al., (1989) Nature 341:544-546, Winter et al., PCT Publication WO90 / 05144A1, which are incorporated herein by reference); and (vi) an isolated complementarity-determining region (CDR). Furthermore, although the two domains of an Fv fragment, VL and VH, are encoded by separate genes, they can be joined by a synthetic linker, allowing them to be produced using recombinant methods as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as a single-chain Fv (scFv); see, e.g., Bird et al. (1988) Science 242:423-426 and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also intended to be encompassed by the term "antigen-binding portion" of an antibody. In certain embodiments, scFv molecules may be incorporated into fusion proteins. Other forms of single-chain antibodies, such as diabodies, are also encompassed.Diabodies are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain but use a linker that is too short to allow pairing between the two domains on the same chain, thereby forcing the domains to pair with the complementary domains on another chain and forming two antigen-binding sites (see, e.g., Holliger, P., et al. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123). Such antibody binding moieties are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790 pp. (ISBN 3-540-41354-5)).

[0030] As used herein, the term "antibody construct" refers to a polypeptide comprising one or more antigen-binding moieties disclosed herein linked to a linker polypeptide or immunoglobulin constant domain. A linker polypeptide comprises two or more amino acid residues linked by a peptide bond and is used to link one or more antigen-binding moieties. Such linker polypeptides are well known in the art (see, e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123). An immunoglobulin constant domain refers to the constant domain of a heavy or light chain. Antibody portions, such as Fab and F(ab')2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion of whole antibodies, respectively. Furthermore, antibodies, antibody portions, and immunoadhesion molecules can be obtained using standard recombinant DNA techniques, as described herein.

[0031] As used herein, an "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to IL1RAP is substantially free of antibodies that specifically bind to antigens other than IL1RAP). However, an isolated antibody that specifically binds to IL1RAP may have cross-reactivity with other antigens, such as IL1RAP molecules from other species. Furthermore, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0032] The term "humanized antibody" refers to an antibody comprising heavy and light chain variable region sequences derived from a non-human species (e.g., mouse), but in which at least a portion of the VH and / or VL sequences has been altered to be more "human-like," i.e., more similar to human germline variable sequences. In particular, the term "humanized antibody" refers to an antibody, or variant, derivative, analog, or fragment thereof, that immunospecifically binds to an antigen of interest and comprises framework (FR) regions having substantially the amino acid sequence of a human antibody and complementarity-determining regions (CDRs) having substantially the amino acid sequence of a non-human antibody. As used herein, the term "substantially" in the context of a CDR refers to a CDR having an amino acid sequence that is at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the amino acid sequence of the non-human antibody CDR. A humanized antibody comprises substantially all of at least one, and typically two, variable domains (Fab, Fab', F(ab')2, FabC, Fv), in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., the donor antibody) and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. Preferably, the humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. In some embodiments, the humanized antibody comprises both a light chain and at least a heavy chain variable domain. The antibody may also include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, the humanized antibody comprises only a humanized light chain. In other embodiments, the humanized antibody comprises only a humanized heavy chain. In specific embodiments, the humanized antibody comprises only humanized variable domains of the light chain and / or humanized heavy chain.

[0033] The humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including but not limited to IgG1, IgG2, IgG3, and IgG4. A humanized antibody may comprise sequences from more than one class or isotype, and particular constant domains can be selected to optimize desired effector functions using techniques well known in the art.

[0034] The terms "Kabat numbering," "Kabat definition," and "Kabat label" are used interchangeably herein. These art-recognized terms refer to a system for numbering amino acid residues that are more variable than other amino acid residues (i.e., hypervariable) in the heavy and light chain variable regions of an antibody or its antigen-binding portion (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391, and Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). In the heavy chain variable region, the hypervariable region ranges from amino acid positions 31 to 35 of CDR1, amino acid positions 50 to 65 of CDR2, and amino acid positions 95 to 102 of CDR3. In the light chain variable region, the hypervariable region ranges from amino acid positions 24-34 for CDR1, amino acid positions 50-56 for CDR2, and amino acid positions 89-97 for CDR3.

[0035] As used herein, the term "CDR" refers to a complementarity-determining region within an antibody variable sequence. Each heavy chain (HC) and light chain (LC) variable region has three CDRs, designated CDR1, CDR2, and CDR3 (or specifically, HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3), respectively. As used herein, the term "CDR set" refers to a group of three CDRs occurring in a single variable region capable of binding an antigen. The exact boundaries of these CDRs are defined differently by different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any antibody variable region, but also provides precise residue boundaries defining the three CDRs. These CDRs are sometimes referred to as Kabat CDRs. Chothia and coworkers (Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987) and Chothia et al., Nature 342:877-883 (1989)) have used the Kabat CDRs to Certain subregions within the CDRs have been found to adopt nearly identical peptide backbone conformations despite great diversity at the amino acid sequence level. These subregions are designated L1, L2, and L3 or H1, H2, and H3, where "L" and "H" indicate the light chain and heavy chain regions, respectively. These regions are sometimes called Chothia CDRs and have boundaries that overlap with the Kabat CDRs. Other boundaries defining CDRs that overlap with the Kabat CDRs have been described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)).Still other CDR boundary definitions may not strictly adhere to one of the above systems, but may be shortened or extended in light of predictions or experimental findings that particular residues or groups of residues, or even entire CDRs, do not significantly affect antigen binding, yet still overlap with the Kabat CDRs. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use the Kabat or Chothia defined CDRs.

[0036] As used herein, the term "framework" or "framework sequence" refers to the remaining sequence of a variable region excluding CDRs. Because the exact definition of a CDR sequence can be determined by different systems, the meaning of a framework sequence is subject to different interpretations accordingly. Furthermore, the six CDRs (CDR-L1, CDR-L2, and CDR-L3 in the light chain and CDR-H1, CDR-H2, and CDR-H3 in the heavy chain) divide the framework regions on the light and heavy chains into four subregions (FR1, FR2, FR3, and FR4) in each chain, where CDR1 is located between FR1 and FR2, CDR2 is located between FR2 and FR3, and CDR3 is located between FR3 and FR4. As referred to by others, the framework region refers to the composite FRs within the variable region of a single naturally occurring immunoglobulin chain, without specifying the specific subregions as FR1, FR2, FR3, or FR4. As used herein, FR refers to one of the four subregions, and FRs refers to two or more of the four subregions that make up a framework region.

[0037] The framework and CDR regions of a humanized antibody need not correspond exactly to the parental sequences. For example, the donor antibody CDR or consensus framework may be mutagenized by substitution, insertion, and / or deletion of at least one amino acid residue such that the CDR or framework residue at that site does not correspond to either the donor antibody or the consensus framework. However, in preferred embodiments, such mutations are not extensive. Typically, at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 95% of the humanized antibody residues correspond to residues in the parental FR and CDR sequences. As used herein, the term "consensus framework" refers to the framework region of a consensus immunoglobulin sequence. As used herein, the term "consensus immunoglobulin sequence" refers to a sequence formed from the amino acids (or nucleotides) that occur most frequently in a family of related immunoglobulin sequences (see, e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987)). In a family of immunoglobulins, each position in the consensus sequence is occupied by the amino acid that occurs most frequently at that position within the family. If two amino acids occur equally frequently, then both can be included in the consensus sequence.

[0038] "Percent (%) amino acid sequence identity" with respect to a peptide or polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a specific peptide or polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment to determine percent amino acid sequence identity can be accomplished in a variety of ways within the skill of those in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. In one embodiment, the present disclosure includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence set forth in any one of SEQ ID NOs: 1-190.

[0039] In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce inflammation. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce autoimmunity. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce a TH-1 mediated immune response. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce a TH2 mediated immune response. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce a TH17 mediated immune response. In one embodiment, the at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, inhibits both a TH-1 mediated immune response and an H-2 mediated immune response. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can reduce the levels of TARC / CCL17, PARC / CCL18, CCL22 / MDC, IgE, periostin, IL-22, IL-13, IL-18, IL-19, CCL27 / CTACK, S100A7 / 12, E-selectin, MMP-12, LDH, neotaxin-1, eotaxin-3 / CCL26, or a combination thereof. In one embodiment, an anti-IL1RAP antibody, such as BFB759, or an antigen-binding portion thereof, can increase or decrease the levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof, or CXCL1, CXCL8, TARC, LCN2, or a combination thereof, toward restoration of normal levels.

[0040] The term "multivalent antibody" is used herein to refer to an antibody that comprises two or more antigen-binding sites. In certain embodiments, multivalent antibodies may be engineered to have three or more antigen-binding sites and are generally not naturally occurring antibodies.

[0041] A "multispecific antibody" refers to an antibody that can bind to two or more unrelated antigens.

[0042] The terms "dual variable domain" or "DVD," as used interchangeably herein, refer to an antigen-binding protein that contains two or more antigen-binding sites and is a tetravalent or multivalent binding protein. Such DVDs can be monospecific, i.e., capable of binding one antigen, or multispecific, i.e., capable of binding two or more antigens. A DVD-binding protein containing two heavy chain DVD polypeptides and two light chain DVD polypeptides is called a DVD Ig. Each half of a DVD Ig contains a heavy chain DVD polypeptide and a light chain DVD polypeptide and two antigen-binding sites. Each binding site contains a heavy chain variable domain and a light chain variable domain, with a total of six CDRs per antigen-binding site involved in antigen binding. In one embodiment, the CDRs described herein are used in an anti-IL1RAP DVD.

[0043] The term "activity" includes activities such as the binding specificity / affinity of an antibody to an antigen, e.g., an anti-hIL1RAP antibody or ADC that binds to the IL1RAP antigen. ADC activity includes binding to IL1RAP in vitro; binding to IL1RAP in vivo on cells expressing IL1RAP; modulating (e.g., inhibiting) IL-1, e.g., IL-1β and / or IL-1α signaling; reducing inflammation; reducing autoimmunity; reducing a TH-1 mediated immune response; reducing a TH2 mediated immune response; reducing levels of TARC / CCL17, PARC / CCL18, CCL22 / MDC, IgE, periostin, IL-22, IL-13, IL-18, IL-19, CCL27 / CTACK, S100A7 / 12, E-selectin, MMP-12, LDH, neotaxin-1, eotaxin-3 / CCL26, or a combination thereof; IL-1α, IL-1β, IL-33, IL-36α, IL- or a combination thereof, or CXCL1, CXCL8, TARC, LCN2, or a combination thereof, toward restoring the levels to normal; alleviating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma; alleviating allergic rhinitis, allergic conjunctivitis, food allergy, drug allergy, or angioedema; or alleviating sepsis, acute respiratory distress syndrome, myocardial infarction, cystic fibrosis, irritable bowel disease, multiple sclerosis, neutrophilic asthma, Alzheimer's disease, stroke, diabetic kidney disease, diabetes, diabetic retinopathy, chronic obstructive pulmonary disease (COPD), or idiopathic pulmonary fibrosis or non-alcoholic fatty liver disease.

[0044] The term "epitope" refers to the region of an antigen bound by an antibody or antibody fragment. In certain embodiments, epitopic determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl, or sulfonyl groups, and may, in certain embodiments, have specific three-dimensional structural characteristics and / or specific drug-binding properties. In certain embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and / or macromolecules.

[0045] As used herein, "k on " or "k a The term "rate constant" is intended to refer to the association rate constant of an antibody to an antigen in forming an antibody / antigen complex.

[0046] As used herein, "k off " or "k d The term "rate constant" is intended to refer to the rate constant of dissociation of an antibody from the antibody / antigen complex.

[0047] As used herein, "K D The term "" is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction. D is k a / k d In one embodiment, an antibody of the invention has a K of about 2,000 nM or less, about 1,000 nM or less, about 500 nM or less, about 200 nM or less, about 100 nM or less, about 75 nM or less, about 25 nM or less, about 21 nM or less, about 12 nM or less, about 11 nM or less, about 10 nM or less, about 9 nM or less, about 8 nM or less, about 7 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, about 1 nM or less, about 0.5 nM or less, about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM or less, or about 0.001 nM or less. D It has.

[0048] As used herein, the term "competitive binding" refers to a situation in which a first antibody competes with a second antibody for a binding site on a third molecule, e.g., an antigen. In one embodiment, competitive binding between two antibodies is determined using FACS analysis.

[0049] The term "competitive binding assay" refers to an assay used to determine whether two or more antibodies bind to the same epitope. In one embodiment, a competitive binding assay is a competitive fluorescence-activated cell sorting (FACS) assay, which is used to determine whether two or more antibodies bind to the same epitope by determining whether the fluorescent signal of a labeled antibody is reduced by the introduction of an unlabeled antibody that competes for the same epitope and reduces the fluorescence level.

[0050] As used herein, the term "labeled antibody" refers to a binding protein, e.g., an antibody, or antigen-binding portion thereof, incorporating a label that provides for the identification of the antibody. Preferably, the label is a detectable label, such as the incorporation of a radiolabeled amino acid or the linkage of a biotinyl moiety to the polypeptide that is detectable by labeled avidin (e.g., streptavidin containing a fluorescent marker or an enzymatic activity that is detectable optically or colorimetrically). Examples of labels for polypeptides include, but are not limited to, radioisotopes or radionuclides (e.g., 3 H, 14 C. 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, or 153Sm), fluorescent labels (e.g., FITC, rhodamine, lanthanide fluorophores), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase), chemiluminescent markers, biotinyl groups, predetermined polypeptide epitopes recognized by a second reporter (e.g., leucine zipper pair sequences, binding sites for second antibodies, metal binding domains, epitope tags), and magnetic agents such as gadolinium chelates.

[0051] The term "antibody-drug conjugate" or "ADC" refers to a binding protein, such as an antibody or antigen-binding fragment thereof, chemically conjugated to one or more chemical drug(s) (also referred to herein as drug(s)), which may optionally be therapeutic or cytotoxic agents. In preferred embodiments, an ADC comprises an antibody, a cytotoxic or therapeutic drug, and a linker that allows for the linkage or conjugation of the drug to the antibody. ADCs typically have in the range of 1 to 8 drugs conjugated to the antibody, including 2, 4, 6, or 8 drug-loaded species. Non-limiting examples of drugs that may be included in an ADC include antimitotic agents, antitumor antibiotics, immunomodulatory agents, gene therapy vectors, alkylating agents, angiogenesis inhibitors, antimetabolites, boron-containing agents, chemoprotectants, hormones, antihormonal agents, corticosteroids, photoactive therapeutic agents, oligonucleotides, radionuclide agents, topoisomerase inhibitors, tyrosine kinase inhibitors, radiosensitizers, and the like.

[0052] The terms "V-set domain-containing T-cell activation inhibitor-1 antibody drug conjugate," "anti-IL1RAP antibody drug conjugate," or "anti-IL1RAP ADC" are used interchangeably herein and refer to an ADC comprising an antibody that specifically binds to IL1RAP, whereby the antibody is conjugated to one or more chemical agent(s) or payload. In one embodiment, the chemical agent(s) is / are attached to the antibody via a linker.

[0053] The term "drug-antibody ratio" or "DAR" refers to the number of drugs, e.g., IGN, auristatins, maytansinoids, linked to the antibodies of an ADC. The DAR of an ADC ranges from 1 to 8, although higher loadings, e.g., 10, are possible depending on the number of binding sites on the antibody. The term DAR may be used in reference to the number of drugs loaded onto an individual antibody, or it may be used in reference to the average or mean DAR of a group of ADCs.

[0054] As used herein, the term "IL1RAP-associated disorder" includes any disorder or disease (including proliferative disorders, e.g., cancer) that is indicated, diagnosed, detected, or identified by phenotypic or genotypic abnormalities in IL1RAP gene components or expression during the course or pathogenesis of the disease or disorder. In this regard, a phenotypic abnormality or determinant of IL1RAP may include, for example, an increase or decrease in the level of IL1RAP protein expression in a cell population, e.g., a cancer cell population, compared to another cell population, e.g., a normal cell population, or an increase or decrease in IL1RAP protein expression in a specific, definable cell population, or an increase or decrease in IL1RAP protein expression during an inappropriate phase or stage of the cellular life cycle. It will be understood that similar expression patterns of genotypic determinants of IL1RAP (e.g., mRNA transcription levels) may also be used to classify or detect IL1RAP-associated disorders.

[0055] As used herein, the term "inflammatory disease" is meant to refer to or describe diseases and disorders characterized by inflammation, an immune response characterized by dilated capillaries, leukocyte infiltration, redness, warmth, pain, and upregulation of inflammatory cytokines, including, but not limited to, interleukin-1 (IL-1), IL-2, IL-6, IL-12, IL-18, tumor necrosis factor alpha (TNF-α), interferon gamma (IFNγ), and granulocyte-macrophage colony-stimulating factor (GM-CSF).

[0056] In some embodiments, the inflammatory disease is characterized by elevated serum levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof, or CXCL1, CXCL8, TARC, LCN2, or a combination thereof.

[0057] As used herein, the term "autoimmune disease" or "autoimmune" is meant to refer to or describe a disease, disorder, or condition characterized by an autoimmune response in which the immune system attacks the body's own healthy cells, tissues, and organs.

[0058] As in the context of neutrophil and eosinophil dysfunction, the term "dysfunction" or grammatical variations thereof refers to cells that are not behaving, working, or operating normally or properly.

[0059] As used herein, the term "increased cell count," such as increased neutrophils or eosinophils, or grammatical variations thereof, refers to abnormal cell counts above non-diseased basal (normal) levels, where increased cell counts are well known in the art as being indicative of disease, such as inflammatory or autoimmune disease.

[0060] As used herein, the term "abnormal serum levels," such as elevated levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or combinations thereof, or grammatical variations thereof, refers to abnormal cytokine levels above non-disease basal (normal) levels, where elevated cytokine levels are well known in the art as being indicative of a disease, such as an inflammatory disease or an autoimmune disease.

[0061] In some embodiments, autoimmune disease or autoimmunity is characterized by the presence of autoantibodies directed against self-antigens in the body's own healthy cells, tissues, or organs.

[0062] In some embodiments, the autoimmune disease or autoimmunity is characterized by elevated serum levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof, or CXCL1, CXCL8, TARC, LCN2, or a combination thereof.

[0063] In some embodiments, the inflammatory or autoimmune disease is sepsis, acute respiratory distress syndrome, myocardial infarction, cystic fibrosis, irritable bowel disease, ulcerative colitis, Crohn's disease, atopic dermatitis, psoriasis, multiple sclerosis, neutrophilic asthma, Alzheimer's disease, stroke, diabetic kidney disease, diabetes, diabetic retinopathy, hidradenitis suppurativa, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, asthma, allergic rhinitis, allergic conjunctivitis, food allergies, drug allergies, as well as angioedema, allergic contact dermatitis, irritant contact dermatitis, rosacea, plaque psoriasis, pustular psoriasis, obesity cell cystitis, systemic lupus erythematosus, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous diseases, Behçet's disease, vitiligo, IL-36 receptor antagonist (DITRA) deficiency, CARD14-mediated psoriasis, acute generalized exanthematous pustulosis, pyoderma gangrenosum, Sweet's syndrome, acne, folliculitis and eosinophilic pustular folliculitis, adult-onset Still's disease, cutaneous lupus erythematosus, contact dermatitis, alopecia areata, cutaneous drug eruptions, graft-versus-host disease, cryopyrin-associated periodic syndrome, granulomatosis with polyangiitis, septic arthritis, pyoderma gangrenosum and acne (PAPA), familial Mediterranean fever, rosacea, synovitis, pustulosis , Bone hyperplasia, Osteitis (SAPHO), Bullous pemphigoid, Pemphigus vulgaris, Schnitzler syndrome, SAPHO, Acne vulgaris, Urticarial vasculitis, Cryopyrin-associated periodic syndromes (CAPS), Hyper IgD syndrome (HIDS) also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), PASH, PFAPA, Generalized pustular psoriasis (GPP), Palmoplantar pustular psoriasis (PPP), Dermatomyositis, Pangranulomatosis, Erdheim-Chester syndrome, Adenosine deaminase deficiency DADA2, Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), A20 haploin deficiency (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), CARD-14-mediated pustular psoriasis (CAMPS), chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, non-alcoholic fatty liver disease, neutrophilic dermatosis,Or a solitary autoinflammatory skin disease.

[0064] As used herein, the term "atopy" or "atopic disease" is meant to refer to or describe a disease, disorder, or condition characterized by a tendency to produce an exaggerated immunoglobulin E (IgE) immune response to a variety of antigens / allergens, including harmless substances.

[0065] In some embodiments, the atopic disease is atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, food allergy, drug allergy, or angioedema. In one embodiment, the atopic disease is atopic dermatitis. The clinical phenotype characterizing atopic dermatitis is skin barrier dysfunction and immune dysregulation. Mutations in the filaggrin gene cause skin barrier dysfunction and transepidermal water loss, leading to atopic dermatitis. Atopic dermatitis is characterized by elevated serum IgE levels and a Th2 immune response accompanied by elevated IL-4, IL-5, IL-10, and IL-13. This leads to increased exposure to allergens, which are taken up by Langerhans cells in lymph nodes and stimulate naive CD4+ T cells (Th0) to differentiate into Th2 cells. Related cytokines, such as IL-4 and IL-13, are produced and are known to stimulate IgE production, while IL-5 is one of the most important cytokines for eosinophil production. IgE mediates mast cell activation, induces keratinocyte expression of inflammatory cytokines, and dendritic cell migration. Impaired barrier function may stimulate signaling cascades leading to epidermal homeostatic responses and type 2 inflammatory responses.

[0066] In one embodiment, an antibody of the invention is administered to a patient with an inflammatory or autoimmune disease, or inflammation or autoimmunity. In one embodiment, an antibody of the invention is administered to a patient with an atopic disorder or disease. In one embodiment, an antibody of the invention is administered to a patient with atopic dermatitis. In one embodiment, administration of an antibody of the invention reduces inflammation or autoimmunity. In one embodiment, administration of an antibody of the invention reduces atopic dermatitis.

[0067] Methods for detecting the expression of IL1RAP are known in the art.

[0068] The terms "overexpress," "overexpression," or "overexpressed" interchangeably refer to a gene that is transcribed or translated at a detectably greater level than normal cells or cells under normal conditions. Thus, overexpression refers to both protein and RNA overexpression (due to transcription, post-transcriptional processing, translation, post-translational processing, changes in stability, and changes in proteolysis), as well as localized overexpression due to changes in protein transport (increased nuclear localization) and enhanced functional activity (e.g., increased substrate hydrolases). Thus, overexpression refers to either the protein level or the RNA level. Overexpression can also be 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to normal cells or comparison cells.

[0069] As used herein, the term "administration" refers to the delivery of a substance (e.g., an anti-IL1RAP antibody) to achieve a therapeutic objective (e.g., treatment of an inflammatory or autoimmune disease or disorder, or reduction of inflammation or autoimmunity, or reduction of atopic disease). Methods of administration can be parenteral, enteral, or topical. Parenteral administration is usually by injection and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, intracutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0070] As used herein, the term "combination therapy" refers to the administration of two or more therapeutic agents, e.g., an anti-IL1RAP antibody and at least one additional therapeutic agent. The additional therapeutic agent may be administered concomitantly with, before, or after anti-IL1RAP administration. In embodiments, an anti-IL1RAP antibody or ADC of the invention is administered in combination with one or more of an inhibitor of a TH2-related immune response, a TH1-related immune response, pruritus, IL-4 signaling, IL-13 signaling, IL-22 signaling, IL-33 signaling, IL-17 signaling, IL-36 signaling, IL-18 signaling, IL-23 signaling, OX40 signaling, IL-5 signaling, T cell migration, IRAK4 signaling, complement signaling, or PDE4 signaling, or a combination thereof, for the treatment of an inflammatory or autoimmune disease, such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma.

[0071] As used herein, the terms "effective amount" or "therapeutically effective amount" refer to an amount of an agent, e.g., an antibody, sufficient to reduce or ameliorate the severity and / or duration of, or one or more symptoms of, a disease, e.g., an inflammatory or autoimmune disease such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma; cause regression of the disorder; prevent the recurrence, development, onset, or progression of one or more symptoms associated with the disorder; detect the disorder; or enhance or improve the prophylactic or therapeutic effect(s) of another therapy (e.g., a prophylactic or therapeutic agent). An effective amount of an antibody may, for example, reduce inflammation (e.g., reduce levels of inflammatory markers), reduce autoimmunity (e.g., reduce autoantibody levels or reduce the immune response against the body's own healthy cells and tissues), decrease the number or size of inflammatory, autoimmune, and / or atopic lesions, and / or provide some relief from one or more symptoms associated with the inflammatory, autoimmune, or atopic disease.

[0072] Various aspects of the invention are described in further detail in the following subsections.

[0073] II. Anti-IL1RAP antibody One aspect disclosed herein provides a humanized anti-IL1RAP antibody, or antigen-binding portion thereof. Another aspect disclosed herein provides a human anti-IL1RAP antibody, or antigen-binding portion thereof. In one embodiment, the antibody disclosed herein binds to human IL1RAP. In another embodiment, the antibody disclosed herein binds to cynomolgus monkey IL1RAP. In another embodiment, the antibody disclosed herein binds to human IL1RAP expressed on a cell.

[0074] In one embodiment, an anti-IL1RAP antibody is disclosed that has the ability to bind to IL1RAP, as described in the Examples below. Collectively, the novel antibodies are referred to herein as "IL1RAP antibodies." The anti-IL1RAP antibody or antigen-binding fragment thereof can inhibit or reduce inflammatory, autoimmune, and / or atopic responses in vivo. In various embodiments, the anti-IL1RAP antibody or antigen-binding fragment thereof can modulate the biological function of IL1RAP. In other embodiments of the foregoing aspects, the anti-IL1RAP antibody or antigen-binding fragment thereof binds to IL1RAP on cells that express IL1RAP. Accordingly, the present disclosure includes an anti-IL1RAP antibody or antigen-binding fragment thereof that is effective in inhibiting or reducing inflammation, autoimmunity, and / or atopy. Without wishing to be bound by any particular theory, in one embodiment, anti-IL1RAP antibodies, antigen-binding portions thereof, and ADCs can inhibit multiple IL1RAP activities, including, but not limited to, IL-1β signaling via IL1RAP, IL-1α, IL-1β, and IL-38 signaling via IL-1R, IL-33 signaling via IL-33R, and IL-36α, IL-36β, and IL-36g signaling via IL-36R.

[0075] Additionally, the present inventors have further demonstrated that anti-IL1RAP antibodies, such as BFB759, are effective in a mouse model of atopic dermatitis (see Example 8). Thus, anti-IL1RAP antibodies and antigen-binding portions thereof can be used to treat atopic dermatitis, or contact dermatitis, allergic dermatitis, or allergic contact dermatitis in a subject.

[0076] Antibodies combining any of the foregoing features are also contemplated as aspects of the present disclosure.

[0077] It should be noted that while the term "antibody" is used throughout, antibody fragments (i.e., anti-IL1RAP antigen-binding portions) are also included in this disclosure and may be included in the embodiments (methods and compositions) described throughout. In certain embodiments, the anti-IL1RAP antibody-binding portion is a Fab, Fab', F(ab'), Fv, disulfide-linked Fv, scFv, single-domain antibody, or diabody.

[0078] Example 2 describes the generation of fully human IL1RAP antibodies directed against the extracellular domain of human IL1RAP, which are contemplated for use in the therapeutic methods described herein. The heavy and light chain variable region amino acid sequences of these human anti-IL1RAP antibodies are shown in Table 5. The heavy and light chain variable region nucleotide sequences of these human antibodies are shown in Table 6.

[0079] Thus, in certain embodiments for use in the methods provided herein, the disclosure provides a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 9, 17, 25, 32, 40, 48, 56, 64, 71, 74, 83, 90, 96, 100, 106, 109, 116, 118, 121, 123, 125, 127, 130, 136, 140, 144, 151, 158, 163, 170, 173, 180, and 185. and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 13, 21, 29, 36, 44, 52, 60, 68, 73, 78, 82, 87, 93, 98, 103, 108, 113, 114, 120, 122, 124, 126, 128, 134, 137, 143, 147, 154, 160, 167, 172, 177, 184, and 189.

[0080] In certain embodiments for use in the methods provided herein, the disclosure includes a human anti-IL1RAP antibody, or antigen-binding portion thereof, comprising a HC CDR set (CDR1, CDR2, and CDR3) selected from those set forth in Table 5, and a LC CDR set (CDR1, CDR2, and CDR3) selected from those set forth in Table 5.

[0081] In certain embodiments, the human anti-IL1RAP antibody for use in the inventive methods herein is selected from 37E10_15B5, 44E5_15C5, 16H2_17D2, and / or 36A10_21B6 (see Table 5).

[0082] In certain embodiments, the human anti-IL1RAP antibody for use in the methods of the invention herein is selected from antibodies including:

[0083] a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 67, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 66, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 70, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 69; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 15, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 51, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 50, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 49, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 55, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 54, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 53; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 59, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 58, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 57, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 63, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 61; and / or BFB759 (i.e., 37E10_15B5) corresponding to a heavy chain variable region comprising a CDR3 domain having the amino acid sequence of SEQ ID NO: 176, a CDR2 domain having the amino acid sequence of SEQ ID NO: 175, and a CDR1 domain having the amino acid sequence of SEQ ID NO: 174, and a light chain variable region comprising a CDR3 domain having the amino acid sequence of SEQ ID NO: 179, a CDR2 domain having the amino acid sequence of SEQ ID NO: 178, and a CDR1 domain having the amino acid sequence of SEQ ID NO: 79.

[0084] In other embodiments, the human anti-IL1RAP antibody for use in the methods of the invention herein is selected from antibodies including: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 68; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 82; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 48 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 52; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 60; or BFB759 (i.e., 37E10_15B5) corresponding to a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 173 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177.

[0085] In a specific embodiment, the human anti-IL1RAP antibody for use in the methods of the invention herein is BFB759 (i.e., 37E10_15B5), which corresponds to a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 67, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 66, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 70, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 69. In another specific embodiment, BFB759 (i.e., 37E10_15B5) corresponds to a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 68.

[0086] Substitution of amino acid residues in the Fc portion of an antibody to alter its effector function has been described (Winter, et al., U.S. Patent Nos. 5,648,260 and 5,624,821, incorporated herein by reference). The Fc portion of an antibody mediates several important effector functions, such as cytokine induction, ADCC, phagocytosis, complement-dependent cytotoxicity (CDC), and half-life / clearance rate of the antibody and antigen-antibody complex. While these effector functions are sometimes desirable for therapeutic antibodies, they may be unnecessary or even harmful depending on the therapeutic purpose. Certain human IgG isotypes, particularly IgG1 and IgG3, mediate ADCC and CDC through binding to FcγRs and complement C1q, respectively. The neonatal Fc receptor (FcRn) is an important component determining the circulating half-life of an antibody. In yet another embodiment, at least one amino acid residue is substituted in the antibody constant region, e.g., the Fc region, to alter the antibody's effector function.

[0087] One embodiment includes a labeled anti-IL1RAP antibody or antibody portion thereof, wherein the antibody is derivatized or conjugated to one or more functional molecule(s) (e.g., another peptide or protein). For example, a labeled antibody can be derivatized by functionally linking (by chemical coupling, genetic fusion, non-covalent attachment, etc.) an antibody or antibody portion of the disclosure to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or protein, diabody, etc.), a detection agent, a pharmaceutical agent, a protein or peptide capable of mediating association of the antibody or antibody portion with another molecule (e.g., a streptavidin core region or a polyhistidine tag), and / or a therapeutic agent selected from the group consisting of an antimitotic agent, an immunomodulatory agent, a gene therapy vector, an alkylating agent, an angiogenesis inhibitor, an antimetabolite, a boron-containing agent, a chemoprotectant, a hormone, an antihormonal agent, a corticosteroid, a photoactive therapeutic agent, an oligonucleotide, a radionuclide agent, a topoisomerase inhibitor, a tyrosine kinase inhibitor, a radiosensitizer, and combinations thereof.

[0088] Another embodiment of the present disclosure provides a glycosylated binding protein, in which an anti-IL1RAP antibody or its antigen-binding portion comprises one or more carbohydrate residues. Nascent in vivo protein production may undergo further processing known as post-translational modification. In particular, sugar (glycosyl) residues may be added enzymatically, a process known as glycosylation. As a result, proteins bearing covalently attached oligosaccharide side chains are known as glycosylated proteins or glycoproteins. Antibodies are glycoproteins, possessing one or more carbohydrate residues in the Fc domain as well as the variable domain. Carbohydrate residues in the Fc domain have a significant impact on the effector function of the Fc domain but have minimal impact on antigen binding or antibody half-life (R. Jefferis, Biotechnol. Prog. 21 (2005), pp. 11-16). In contrast, glycosylation of the variable domain may affect the antigen-binding activity of an antibody. Glycosylation of variable domains can have a negative effect on antibody binding affinity, possibly due to steric hindrance (Co, MS, et al., Mol. Immunol. (1993) 30:1361-1367) or increased affinity for antigen (Wallick, SC, et al., Exp. Med. (1988) 168:1099-1109; Wright, A., et al., EMBO J. (1991) 10:2717-2723).

[0089] One embodiment of the present disclosure is directed to the creation of glycosylation site mutants in which the O- or N-linked glycosylation site of a binding protein is mutated. Those skilled in the art can create such mutants using standard, well-known techniques. Glycosylation site mutants that retain biological activity but have increased or decreased binding activity are another object of the present disclosure.

[0090] Expression of glycosylated proteins different from those of host cells can be achieved by genetically modifying the host cells to express heterologous glycosylation enzymes. Recombinant techniques can be used to generate antibodies or antigen-binding portions thereof that exhibit glycosylation of human proteins. For example, yeast strains have been genetically engineered to express non-naturally occurring glycosylation enzymes so that glycosylated proteins (glycoproteins) produced in these yeast strains exhibit protein glycosylation identical to that of animal cells, particularly human cells (U.S. Patent Publication Nos. 20040018590 and 20020137134, PCT Publication WO2005100584A2).

[0091] Antibodies may be produced by any of a number of techniques, including expression from a host cell, in which the expression vector(s) encoding the heavy and light chains are transfected into the host cell by standard techniques. The various forms of the term "transfection" are intended to encompass various techniques commonly used for introducing foreign DNA into prokaryotic or eukaryotic host cells, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection, etc. Although antibodies can be expressed in either prokaryotic or eukaryotic host cells, expression in eukaryotic host cells is preferred, and expression in mammalian host cells is most preferred, because such eukaryotic cells, particularly mammalian cells, are more likely than prokaryotic cells to assemble and secrete properly folded and immunologically active antibodies.

[0092] Preferred mammalian host cells for expressing the recombinant antibodies disclosed herein include Chinese hamster ovary (CHO) cells (described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, including dhfr-CHO cells used with the DHFR selectable marker as described, e.g., in R.J. Kaufman and PA Sharp (1982) Mol. Biol. 159:601-621), NSO myeloma cells, COS cells, and SP2 cells. When a recombinant expression vector encoding an antibody gene is introduced into a mammalian host cell, the antibody is produced by culturing the host cell for a period of time sufficient to allow for expression of the antibody in the host cell, or more preferably, secretion of the antibody into the culture medium in which the host cell is grown. The antibody can be recovered from the culture medium using standard protein purification methods.

[0093] Host cells can also be used to produce functional antibody fragments, such as Fab fragments or scFv molecules. It will be understood that variations on the above procedures are within the scope of the present disclosure. For example, it may be desirable to transfect host cells with DNA encoding functional fragments of either the antibody light and / or heavy chains. Recombinant DNA technology may also be used to remove some or all of the DNA encoding either or both of the light and heavy chains that is not necessary for binding to the antigen of interest. Molecules expressed from such truncated DNA molecules are also encompassed by the antibodies of the present disclosure. Additionally, antibodies of the present disclosure may be crosslinked to a second antibody using standard chemical crosslinking methods to produce bifunctional antibodies in which one heavy chain and one light chain are an antibody of the present disclosure and the other heavy and light chains are specific for an antigen other than the antigen of interest.

[0094] In a preferred system for recombinant expression of an antibody or its antigen-binding portion, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into CHO cells constituting a glutamine synthetase expression system, which is commercially available from Lonza (hereafter referred to as GS-CHO) (Bebbington, CR et al. (1992), Biotechnology, 10, pages 169-175). In another system for recombinant expression of an antibody or its antigen-binding portion, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into dhfr-CHO cells by calcium phosphate-mediated transfection. Within the recombinant expression vector, the antibody heavy chain and antibody light chain genes are each operably linked to a CMV enhancer / AdMLP promoter control element, driving high-level transcription of the genes. The recombinant expression vector also contains a DHFR gene, allowing for selection of CHO cells transfected with the vector using methotrexate selection / amplification. The selected transformed host cells are cultured to express the antibody heavy and light chains, and intact antibody is recovered from the culture medium. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells, and recover the antibody from the culture medium. The present disclosure further provides a method for synthesizing a recombinant antibody by culturing host cells in an appropriate culture medium until the recombinant antibody is synthesized. Recombinant antibodies may be produced using nucleic acid molecules corresponding to the amino acid sequences disclosed herein. In one embodiment, nucleic acid molecules set forth in SEQ ID NOs: 191-259 are used to produce the recombinant antibody. The method may further include isolating the recombinant antibody from the culture medium.

[0095] III. Anti-IL1RAP Antibody-Drug Conjugates (ADCs) The anti-IL1RAPs described herein may be conjugated to a drug moiety to form an anti-IL1RAP antibody-drug conjugate (ADC). Antibody-drug conjugates (ADCs) may enhance the therapeutic efficacy of antibodies in treating disease due to the ability of the ADC to selectively deliver one or more drug moieties to target tissues or cells. Thus, in certain embodiments, the present disclosure provides anti-IL1RAP ADCs for therapeutic use.

[0096] Anti-IL1RAP ADCs comprise an anti-IL1RAP antibody, i.e., an antibody that specifically binds to IL1RAP, conjugated to one or more drug moieties. The specificity of the ADC is defined by the specificity of the antibody, i.e., anti-IL1RAP.

[0097] Examples of linkers that can be used to conjugate an antibody to one or more drug(s) in anti-IL1RAP ADCs are shown below. The terms "drug," "agent," and "drug moiety" are used interchangeably herein. The terms "linked" and "conjugated" are also used interchangeably herein to refer to the covalent attachment of an antibody to a moiety.

[0098] In some embodiments, the ADC has the following formula (Formula I): [ka] wherein Ab is an anti-IL1RAP antibody described herein, (LD) is a linker-drug moiety. The linker-drug moiety is made up of a linker, L-, and a drug moiety, -D, that has cytostimulatory, cytotoxic, or other therapeutic activity against a target cell, e.g., a cell expressing IL1RAP, and n is an integer from 1 to 20. In some embodiments, n is 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. The DAR of the ADC corresponds to "n" as referred to in Formula I.

[0099] Additional details regarding drugs (D in Formula I) and linkers (L in Formula I) that may be used in ADCs, as well as alternative ADC structures, are described in U.S. Patent No. 11,248,054, which is incorporated herein by reference in its entirety for all purposes.

[0100] Conjugation of a drug to an antibody via a linker can be achieved by any technique known in the art. Many different reactions are available for covalently linking the drug and linker to the antibody. This can be achieved by reaction of amino acid residues on the antibody, such as the amine groups of lysine, the free carboxylic acid groups of glutamic and aspartic acids, the sulfhydryl groups of cysteine, or various sites on aromatic amino acids. The most commonly used nonspecific method of covalent attachment is the carbodiimide reaction, in which a carboxyl (or amino) group of a compound is linked to an amino (or carboxy) group of an antibody. Additionally, bifunctional agents such as dialdehydes and imidoesters have been used to link amino groups of compounds to amino groups of antibodies. Another method for linking drugs to antibodies is the Schiff base reaction. This method involves oxidizing a drug containing a glycol or hydroxyl group with periodate, resulting in the formation of an aldehyde, which can then react with the linking agent. Linkage occurs by forming a Schiff base with the amino group of the antibody. Isothiocyanates can also be used as coupling agents to covalently link agents to antibodies. Other techniques are well known to those of skill in the art and are within the scope of this disclosure.

[0101] In certain embodiments, an intermediate that is a precursor to the linker is reacted with the drug under appropriate conditions. In certain embodiments, a reactive group is used on the drug or the intermediate. The reaction product of the drug and intermediate, or the derivatized drug, is then reacted with the anti-IL1RAP antibody under appropriate conditions. The synthesis and structures of exemplary linkers, stretcher units, amino acid units, and autoimmune spacer units are described in U.S. Patent Application Publication Nos. 20030083263, 20050238649, and 20050009751, each of which is incorporated herein by reference.

[0102] The stability of the ADC may be measured by standard analytical techniques such as mass spectrometry, HPLC, and separation / analysis techniques such as LC / MS.

[0103] IV. Methods of Use for Anti-IL1RAP Antibodies Anti-IL1RAP antibodies, such as BFB759, and antibody portions (and ADCs) can neutralize human IL1RAP activity both in vivo and in vitro. Thus, such antibodies and antibody portions can be used to inhibit hIL1RAP activity, for example, in cell cultures containing hIL1RAP, in human subjects, or in other mammalian subjects having an IL1RAP with which the antibodies disclosed herein cross-react. In one embodiment, the present disclosure provides a method of inhibiting hIL1RAP activity, comprising contacting hIL1RAP with an antibody, such as BFB759, or antibody portion, such that hIL1RAP activity is inhibited. For example, in a cell culture containing or suspected of containing hIL1RAP, an anti-IL1RAP antibody, such as BFB759, or antibody portion can be added to the culture medium to inhibit hIL1RAP activity in the culture.

[0104] In another embodiment, disclosed herein is a method of reducing hIL1RAP activity in a subject in need thereof, wherein the subject is suffering from an inflammatory, autoimmune, or atopic disease or disorder, such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, and asthma, or a disorder in which IL1RAP activity is detrimental. The present disclosure provides a method for reducing IL1RAP activity in a subject suffering from such a disease or disorder, the method comprising administering to the subject an anti-IL1RAP antibody or antibody portion of the disclosure, such that IL1RAP activity in the subject is reduced.

[0105] Preferably, the IL1RAP is human IL1RAP and the subject is a human subject. Alternatively, the subject can be a mammal expressing an IL1RAP capable of binding by an antibody of the present disclosure. Furthermore, the subject can be a mammal into which IL1RAP has been introduced (e.g., by administration of IL1RAP or expression of an IL1RAP transgene). Anti-IL1RAP antibodies of the present disclosure can be administered to a human subject for therapeutic purposes. Furthermore, anti-IL1RAP antibodies of the present disclosure can be administered to a non-human mammal expressing an IL1RAP capable of binding to the antibody for veterinary purposes or as an animal model of human disease. Regarding the latter, such animal models can be useful for evaluating the therapeutic efficacy (e.g., testing efficacy, dosage, and administration time courses) of the antibodies and ADCs of the present disclosure.

[0106] As used herein, the term "disorder in which IL1RAP activity is detrimental" is intended to include diseases and other disorders in which the presence of IL1RAP in a subject suffering from the disorder has been shown or is suspected to be a contributing factor to the pathophysiology of the disorder or to the aggravation of the disorder. Thus, a disorder in which IL1RAP activity is detrimental is one in which reducing IL1RAP activity is expected to alleviate the symptoms and / or progression of the disorder. Such a disorder may be evidenced, for example, by an increased concentration of IL1RAP in the biological cells, fluids, or tissues of a subject suffering from the disorder (e.g., an increased concentration of IL1RAP in a subject's tumor, serum, plasma, synovial fluid, etc.), which can be detected, for example, using anti-IL1RAP as described above.

[0107] Non-limiting examples of diseases and disorders that can be treated with an anti-IL1RAP antibody, such as BFB759, or an antigen-binding fragment thereof according to the present invention include the following inflammatory diseases, autoimmune diseases, and atopic diseases and disorders. For example, suitable diseases and disorders include sepsis, acute respiratory distress syndrome, myocardial infarction, cystic fibrosis, irritable bowel disease, ulcerative colitis, Crohn's disease, atopic dermatitis, psoriasis, multiple sclerosis, neutrophilic asthma, Alzheimer's disease, stroke, diabetic nephropathy, diabetes, diabetic retinopathy, hidradenitis suppurativa, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, asthma, allergic rhinitis, allergic conjunctivitis, food allergies, drug allergies, as well as angioedema, allergic contact dermatitis, irritant contact dermatitis, and the like. Rosacea, plaque psoriasis, pustular psoriasis, mastocytosis, systemic lupus erythematosus, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous diseases, Behçet's disease, vitiligo, IL-36 receptor antagonist (DITRA) deficiency, CARD14-mediated psoriasis, acute generalized exanthematous pustulosis, pyoderma gangrenosum, Sweet's syndrome, acne, folliculitis and eosinophilic pustular folliculitis, adult-onset Still's disease, cutaneous lupus erythematosus, contact dermatitis, alopecia areata, cutaneous drug eruptions, graft-versus-host disease, cryopyrin-associated periodic syndrome, polyangiitis Blastocystosis, septic arthritis, pyoderma gangrenosum and acne (PAPA), familial Mediterranean fever, rosacea, synovitis, pustulosis, hyperostosis, osteitis (SAPHO), bullous pemphigoid, pemphigus vulgaris, Schnitzler syndrome, SAPHO, acne vulgaris, urticarial vasculitis, cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndrome (TRAPS), IL-1 receptor antagonist deficiency (DIRA), PASH , PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis, pangranulomatosis, Erdheim-Chester syndrome, adenosine deaminase deficiency (DADA2), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), A20 haploinsufficiency (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), CARD-14-mediated pustular psoriasis (CAMPS), chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC),These include, but are not limited to, pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, nonalcoholic fatty liver disease, neutrophilic dermatosis, or isolated autoinflammatory skin disease.

[0108] In a specific embodiment, the anti-IL1RAP antibody is BFB759 and is used to treat atopic dermatitis. In another embodiment, the anti-IL1RAP antibody is BFB759 and is used to treat irritable bowel syndrome, Crohn's disease, or ulcerative colitis. In another embodiment, the anti-IL1RAP antibody is BFB759 and is used to treat asthma.

[0109] In one embodiment, the anti-IL1RAP antibodies disclosed herein, such as BFB759, are used to treat inflammation, autoimmunity, or atopy. In another embodiment, the anti-IL1RAP antibodies disclosed herein, such as BFB759, and ADCs are used to treat inflammatory, autoimmune, or atopic diseases. The diseases and disorders described herein may be treated with anti-IL1RAP antibodies or ADCs, as well as pharmaceutical compositions comprising such anti-IL1RAP antibodies or ADCs.

[0110] In certain embodiments, the antibodies and ADCs disclosed herein are administered to a subject in need thereof to treat an inflammatory, autoimmune, or atopic disease, disorder, or condition that exhibits, or is likely to exhibit, elevated levels of IL1RAP.

[0111] In certain embodiments, the present disclosure includes a method for treating (e.g., curing, suppressing, ameliorating, delaying or preventing onset, or preventing recurrence or relapse) an inflammatory or autoimmune disease, disorder, or condition in a mammal in need thereof, comprising administering a therapeutically effective amount of an anti-IL1RAP antibody against IL1RAP described herein, such as BFB759. In further embodiments, the inflammatory or autoimmune condition is an atopic condition, such as atopic dermatitis, and the anti-IL1RAP antibody, such as BFB759, reduces the level of TARC / CCL17 (a marker) at the atopic site or in the serum. In further embodiments, the inflammatory or autoimmune condition is characterized by or caused by neutrophil or eosinophil dysfunction. In further embodiments, the inflammatory or autoimmune condition is further characterized by suppurative inflammation. In a further embodiment, the antibody is characterized by inhibition of IL-8 release by intestinal epithelial cells, intestinal myofibroblasts, or skin fibroblasts stimulated with IL-1α, IL-1β, IL-36α, IL-36β, and IL-36γ. In a further embodiment, the antibody is administered topically, intradermally, subcutaneously, or intravenously at a dose of 1-20 mg / kg. In a further embodiment, the antibody is administered as a bolus, less than once weekly, twice weekly, more than twice weekly, or by continuous infusion.

[0112] In a further embodiment, the method of treatment further comprises identifying the mammal in need thereof according to abnormal serum levels of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof, or abnormal serum levels of CXCL1, CXCL8, TARC, LCN2, or a combination thereof, hi a further embodiment, the method of treatment results in restoration of these markers to or toward normal levels.

[0113] In a further embodiment, the method of treatment results in a reduction in inflammation, autoimmunity, or atopy as determined by a reduction in the level of inflammatory, autoimmune, or atopic markers known in the art, hi a further embodiment, the method of treatment results in a reduction or elimination of one or more symptoms of an inflammatory, autoimmune, or atopic disease, disorder, or condition.

[0114] In some embodiments, the anti-IL1RAP antibody or fragment thereof used in the methods of the invention is a human or humanized anti-IL1RAP antibody or fragment thereof. In one embodiment, the antibody or antigen-binding portion thereof of the invention comprises an isotype that lacks effector function (e.g., human IgG4).

[0115] Anti-IL1RAP antibodies, such as BFB759, or ADCs, or antigen-binding portions thereof, can be used alone or in combination to treat such diseases. It should be understood that anti-IL1RAP antibodies, such as BFB759, or antigen-binding portions thereof, can be used alone or in combination with at least one additional agent, e.g., a therapeutic agent, which will be selected by those skilled in the art depending on the intended purpose. For example, the additional agent can be a therapeutic agent recognized by those skilled in the art as useful for treating the disease or condition being treated by the antibody. The additional agent can also be an agent that imparts beneficial attributes to the therapeutic composition, such as an agent that affects the viscosity of the composition. The additional agent can be a therapeutic antibody, a small molecular weight molecule, an siRNA, an mRNA, or any other modality known to those skilled in the art.

[0116] It should further be understood that the combinations included in this disclosure are combinations useful for their intended purpose. The agents listed below are exemplary for purposes and are not intended to be limiting. The combinations that are part of this disclosure can be an antibody of this disclosure and at least one additional agent selected from the list below. The combination can also include one or more additional agents, for example, two or three additional agents, provided that the combination allows the formed composition to perform its intended function.

[0117] Combination therapy can include one or more IL1RAP antagonists, e.g., anti-IL1RAP antibodies, such as BFB759, or fragments thereof, formulated and / or co-administered with one or more additional therapeutic agents, e.g., one or more inhibitors of a TH2-related immune response, a TH1-related immune response, a TH17-related immune response, pruritus, IL-4 signaling, IL-13 signaling, IL-22 signaling, IL-33 signaling, IL-17 signaling, IL-36 signaling, IL-18 signaling, IL-23 signaling, OX40 signaling, IL-5 signaling, T cell migration, IRAK4 signaling, complement signaling, PDE4 signaling, or combinations thereof.

[0118] Provided herein are methods for treating inflammatory, autoimmune, and atopic diseases, disorders, and conditions in a patient, comprising administering to the patient an anti-IL1RAP antibody, such as BFB759, or a fragment thereof, or an ADC of the invention, in combination with at least one additional therapeutic agent, wherein the combination therapy exhibits a synergistic effect, e.g., therapeutic synergy, in the subject. As used herein, "synergy" or "therapeutic synergy" refers to the phenomenon in which treatment of a patient with a combination of therapeutic agents exhibits therapeutically superior results than those achieved by using each component of the combination at its optimal dose (Corbett, T.H. et al., Cancer Treatment Reports, 66:1187 (1982)). For example, a therapeutically superior outcome is one in which the patient a) experiences a therapeutic effect that is equal to or greater than that achieved when each component of the combination therapy is administered as a monotherapy at the same dose as the combination therapy, while experiencing fewer adverse events, or b) experiences a therapeutic effect that is greater than that achieved when each component of the combination(s) is administered at the same dose as when administered as an individual component, while experiencing no dose-limiting toxicities.

[0119] In certain embodiments, an anti-IL1RAP antibody, such as BFB759, can be administered alone or with at least one other therapeutic agent that acts in combination or synergistically with the antibody to treat an inflammatory, autoimmune, or atopic disease, disorder, or condition.

[0120] In some embodiments, at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as, for example, BFB759, inhibits a TH2-related immune response. In some embodiments, at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as, for example, BFB759, inhibits a TH1-related immune response. In some embodiments, at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as, for example, BFB759, inhibits a TH17-related immune response. In some embodiments, at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as, for example, BFB759, inhibits both a TH1- and a TH2-related immune response. In some embodiments, at least one therapeutic agent used in combination with an anti-IL1RAP antibody, such as, for example, BFB759, inhibits pruritus.

[0121] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-4 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-4Rα inhibitor or antagonist, a Pan-JAK inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is dupilumab, CBP-201, AK120, celdulatinib, CEE321, jactinib, delgocitinib, filgotinib, PF-06651600, tofacitinib, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods of treating atopic diseases, hi some embodiments, such combinations are administered in methods of treating atopic dermatitis.

[0122] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-13 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-13 inhibitor or antagonist, a JAK1 / JAK2 inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is tralokinumab, lebrikizumab, ASLAN004, baricitinib, ruxolitinib, filgotinib, PF-06651600, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0123] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-22 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-22 inhibitor or antagonist, an IL-22R inhibitor or antagonist, a JAK1 / TYK2 inhibitor or antagonist, a JAK1 / JAK3 inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is fezakinumab, LEO138559, brepositinib, ATI-1777, deuclavatinib, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0124] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-33 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-33 or IL-33R inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is etokimab, REGN3500, astegolimab, PF-06817024, MEDI3506, CNTO7160, or a combination thereof. In some embodiments, such a combination is administered in a method for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such a combination is administered in a method for treating an atopic disease. In some embodiments, such a combination is administered in a method for treating atopic dermatitis. In some embodiments, such combinations are administered in methods of treating psoriasis, hi some embodiments, such combinations are administered in methods of treating allergic contact dermatitis, irritant contact dermatitis, rosacea, plaque psoriasis, pustular psoriasis, mastocytosis, systemic lupus erythematosus, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous diseases, Behcet's disease, or vitiligo.

[0125] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-17 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-17A inhibitor or antagonist, or an IL-17RA inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is secukinumab, ixekizumab, brodalumab, bimekizumab, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease.

[0126] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-36 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-36 inhibitor or antagonist, or an IL-36R inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is spesolimab, imsidolimab, REGN6490, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis. In some embodiments, such combinations are administered in methods for treating neutrophilic dermatosis or hidradenitis pustularis. In some embodiments, such combinations are administered in methods of treating generalized pustular psoriasis, palmoplantar pustular psoriasis, IL-36 receptor antagonist (DITRA) deficiency, plaque psoriasis, CARD14-mediated psoriasis, acute generalized exanthematous pustulosis, hidradenitis suppurativa, pyoderma gangrenosum, Sweet's syndrome, systemic lupus erythematosus, systemic sclerosis, autoimmune bullous diseases, acne, atopic dermatitis, allergic contact dermatitis, or folliculitis and eosinophilic pustular folliculitis.

[0127] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-18 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-18 inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is tadequinig alfa. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis. In some embodiments, such combinations are administered in methods for treating adult-onset Still's disease, lupus erythematosus, or psoriasis. In some embodiments, such combinations are administered in methods to treat adult-onset Still's disease, cutaneous lupus erythematosus, psoriasis, atopic dermatitis, chronic spontaneous urticaria, contact dermatitis, alopecia areata, cutaneous drug eruption, graft-versus-host disease, cryopyrin-associated periodic syndrome, granulomatosis with polyangiitis, systemic sclerosis, hidradenitis suppurativa, septic arthritis, pyoderma gangrenosum and acne (PAPA), familial Mediterranean fever, rosacea, synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO), bullous pemphigoid, pemphigus vulgaris, Behcet's disease, or Schnitzler's syndrome.

[0128] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-23 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-23 inhibitor or antagonist, an IL-12 / 23p40 subunit inhibitor or antagonist, an IL-23p19 subunit inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is risankizumab, ustekinumab, guselkumab, tildrakizumab, mirikizumab, brazikumab, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0129] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits OX40 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an OX40 or OX40L inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is KHK4083, GBR830, KY1005, or a combination thereof. In some embodiments, such a combination is administered in a method for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such a combination is administered in a method for treating an atopic disease. In some embodiments, such a combination is administered in a method for treating atopic dermatitis.

[0130] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits IL-5 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-5Rα inhibitor or antagonist, a JAK1 inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is benralizumab, upadacitinib, abrocitinib, SHR0302, filgotinib, PF-06651600, or a combination thereof. In some embodiments, such a combination is administered in a method for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such a combination is administered in a method for treating an atopic disease. In some embodiments, such a combination is administered in a method for treating atopic dermatitis.

[0131] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits T cell migration. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an S1PR1 inhibitor or antagonist, an S1PR4 inhibitor or antagonist, an S1PR5 inhibitor or antagonist, a CCR4 inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is etrasimod, ozanimod, SCD-044, LC51-0255, BMS-986166, RPT193, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating an atopic disease. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0132] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits pruritus. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-1α inhibitor or antagonist, an OSMRβ inhibitor or antagonist, an NK1R inhibitor or antagonist, a P2X3 inhibitor or antagonist, an IL-31 inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is bermekimab, bicsarelimab, serlopitant, tradipitant, BLU-5937, nemolizumab, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods of treating atopic diseases, hi some embodiments, such combinations are administered in methods of treating atopic dermatitis.

[0133] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits a TH-1-related immune response. In some embodiments, the therapeutic molecule used in combination with an anti-IL1RAP antibody, such as BFB759, is an IL-1α inhibitor or antagonist, an IL-1β inhibitor or antagonist, an IL-1R1 inhibitor or antagonist, an IL-36R inhibitor or antagonist, a TNFα inhibitor or antagonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is bermekimab, anakinra, canakinumab, gevokizumab, rilonacept, MEDI8968, spesolimab, imsidolimab, REGN6490, adalimumab, infliximab, etanercept, certolizumab, golimumab, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating atopic diseases. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.In some embodiments, such combinations are selected from the group consisting of adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, familial Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, acne vulgaris (PAPA), cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis, panniculitis, Erdheim-Chester disease syndrome, adenosine deaminase deficiency (DADA2), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), atopic dermatitis, allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), or NLRC4-associated macrophage activation syndrome (NLRC4-MAS). In some embodiments, such combinations are administered in methods of treating neutrophilic dermatitis, pustular hidradenitis, a monofocal autoinflammatory skin disease, or psoriasis.

[0134] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic molecule that inhibits IRAK4 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is an IRAK4 inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is PF-06650833, CA-4948, KT-474, or a combination thereof. In some embodiments, such a combination is administered in a method for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such a combination is administered in a method for treating an atopic disease. In some embodiments, such a combination is administered in a method for treating atopic dermatitis.

[0135] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits complement signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is a C5a inhibitor or antagonist, a C5aR inhibitor or antagonist, a TSLP inhibitor or antagonist, a CD80 / CD86 inhibitor or antagonist, an IL-6 inhibitor or antagonist, a CD20 inhibitor or antagonist, an integrin α4 inhibitor or antagonist, an AhR agonist, or a combination thereof. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is vilobelimab, FX002, INF904, tezepelumab, abatacept, tocilizumab, sarilumab, rituximab, vedolizumab, tapinarofibrate, tadekinig alfa, or a combination thereof. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating atopic diseases. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0136] In some embodiments, an anti-IL1RAP antibody, such as BFB759, is administered in combination with at least one therapeutic agent that inhibits PDE4 signaling. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is a PDE4 inhibitor or antagonist. In some embodiments, the therapeutic agent used in combination with an anti-IL1RAP antibody, such as BFB759, is apremilast, crisaborole, difamilast, roflumilast, or a combination thereof. In some embodiments, such a combination is administered in a method for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such a combination is administered in a method for treating an atopic disease. In some embodiments, such a combination is administered in a method for treating atopic dermatitis.

[0137] Anti-IL1RAP antibodies, such as BFB759, may be administered in combination with multiple therapeutic agents with different mechanisms of action, such as an agent that inhibits or antagonizes IL-33 and an agent that inhibits or antagonizes IL-36. Any of the molecules or classes of molecules described above may be used in such combinations. In some embodiments, such combinations are administered in methods for treating rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, atopic dermatitis, or asthma. In some embodiments, such combinations are administered in methods for treating atopic diseases. In some embodiments, such combinations are administered in methods for treating atopic dermatitis.

[0138] In some embodiments related to Examples 8 and 9 herein, methods are provided for treating atopic dermatitis or asthma in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with dupilumab. Also provided are methods for treating a disease responsive to lowering serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with dupilumab. Also provided are methods for lowering serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with dupilumab. In some embodiments, diseases contemplated for treatment with the combination of BFB759 and dupilumab herein include, among others, atopic dermatitis, COPD, bronchial asthma, allergic rhinitis, eosinophilic pneumonia, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, mastocytosis, and / or eosinophil-associated diseases.

[0139] In some embodiments related to Example 10 herein, a method of treating atopic dermatitis or asthma in a patient in need thereof is provided, the method comprising administering a therapeutically effective amount of BFB759 in combination with tralokinumab. Also provided is a method of treating a disease responsive to reducing serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with tralokinumab. Also provided is a method of reducing serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with tralokinumab. In some embodiments, diseases contemplated for treatment with the combination of BFB759 and tralokinumab herein include, among others, atopic dermatitis, COPD, bronchial asthma, allergic rhinitis, eosinophilic pneumonia, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, mastocytosis, and eosinophil-associated diseases.

[0140] In some embodiments related to Example 11 herein, there is provided a method of treating atopic dermatitis or asthma in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with one or more of abrocitinib, upadacitinib, dexamethasone, triamcinolone, and / or prednisone. Also provided is a method of treating a disease responsive to lowering serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with one or more of abrocitinib, upadacitinib, dexamethasone, triamcinolone, and / or prednisone. Also provided is a method for lowering serum TARC levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with one or more of abrocitinib, upadacitinib, dexamethasone, triamcinolone, and / or prednisone. In some embodiments, diseases contemplated for treatment herein with BFB759 in combination with abrocitinib, upadacitinib, dexamethasone, triamcinolone and / or prednisone include, among others, atopic dermatitis, COPD, bronchial asthma, allergic rhinitis, eosinophilic pneumonia, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, mastocytosis, and eosinophil-associated diseases.

[0141] In some embodiments related to Examples 12 and 14 herein, a method of treating atopic dermatitis or asthma in a patient in need thereof is provided, the method comprising administering BFB759 in combination with adalimumab. Also provided is a method of treating a disease responsive to lowering serum IL-6 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with adalimumab. Also provided is a method of lowering serum IL-6 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with adalimumab. Also provided is a method of treating a disease responsive to lowering serum IL-6 and IL-8 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with adalimumab. Also provided is a method of lowering serum IL-6 and IL-8 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with adalimumab. Also provided is a method of lowering serum IL-6 and IL-8 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with adalimumab. In some embodiments, diseases contemplated for treatment herein with the combination of BFB759 and adalimumab include, among others, rheumatoid arthritis, systemic juvenile idiopathic arthritis (sJIA), Castleman's disease, giant cell arteritis, Takayasu's arteritis and cytokine release syndrome, adult-onset Still's disease, ankylosing spondylitis, Crohn's disease, ulcerative colitis, hidradenitis suppurativa, psoriasis vulgaris, psoriatic arthritis, inflammatory bowel disease, neutrophilic dermatosis-pyoderma gangrenosum, Behcet's disease, granulomatosis with polyangiitis (also known as Wegener's granulomatosis), sarcoidosis, pemphigus, multicentric reticulohistiocytosis, alopecia areata, inflammatory bowel disease, and / or palmoplantar pustulosis (PPP).

[0142] In some embodiments related to Example 13 herein, there is provided a method of treating atopic dermatitis or asthma in a patient in need thereof, the method comprising administering BFB759 in combination with secukinumab. Also provided is a method of treating a disease responsive to lowering serum IL-6 and IL-8 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with secukinumab. Also provided is a method of lowering serum IL-6 and IL-8 levels in a patient in need thereof, the method comprising administering a therapeutically effective amount of BFB759 in combination with secukinumab. In some embodiments, diseases contemplated for treatment herein with the combination of BFB759 and secukinumab include, among others, rheumatoid arthritis, systemic juvenile idiopathic arthritis (sJIA), Castleman's disease, giant cell arteritis, Takayasu's arteritis and cytokine release syndrome, adult-onset Still's disease, ankylosing spondylitis, Crohn's disease, ulcerative colitis, hidradenitis suppurativa, psoriasis vulgaris, psoriatic arthritis, neutrophilic dermatosis-pyoderma gangrenosum, Behcet's disease, granulomatosis with polyangiitis (also known as Wegener's granulomatosis), sarcoidosis, pemphigus, multicentric reticulohistiocytosis, alopecia areata, inflammatory bowel disease, and / or palmoplantar pustulosis (PPP).

[0143] In other embodiments of the combination methods of the invention provided herein that utilize human anti-hIL1RAP antibodies in combination with other therapeutic agents, other human anti-hIL1RAP antibodies known in the art are also contemplated for use herein.

[0144] A pharmaceutical composition may include a "therapeutically effective amount" or a "prophylactically effective amount" of an antibody or antibody portion. A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result. A therapeutically effective amount of an antibody or antibody portion may be determined by one of skill in the art and may vary depending on factors such as the individual's disease state, age, sex, and weight, and the ability of the antibody or antibody portion to elicit a desired response in the individual. A therapeutically effective amount is also an amount in which any toxic or adverse effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired prophylactic result. Generally, a prophylactic dose is used in subjects at a pre-stage or earlier stage of disease, and therefore the prophylactically effective amount will be less than the therapeutically effective amount.

[0145] Dosage regimens may be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dosage may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. In other embodiments, the anti-IL1RAP antibody, e.g., BFB759, and / or therapeutic agent is administered as a subcutaneous injection or intravenously as a bolus less frequently than once a week, twice a week, more frequently than twice a week, or by continuous infusion. It is contemplated herein that parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, dosage unit form refers to physically discrete units suitable as unitary dosages for the mammalian subject to be treated, each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit forms are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the technology of compounding such active compounds for the treatment of hypersensitivity in an individual.

[0146] In certain embodiments, the effective amount of a pharmaceutical composition comprising a therapeutically employed anti-IL1RAP antibody, such as BFB759, with or without at least one additional therapeutic agent will vary, for example, depending on the therapeutic context and purpose. Thus, one of skill in the art will understand that appropriate dosage levels for treatment with certain embodiments will vary depending, in part, on the indication for which the delivered molecule, anti-IL1RAP antibody, such as BFB759, with or without at least one additional therapeutic agent, is being used, the route of administration, and the patient's size (weight, body surface, or organ size) and / or condition (age and general health). In certain embodiments, a clinician can titrate and modify the route of administration to obtain optimal therapeutic effects. In certain embodiments, typical dosages can range from about 0.1 μg / kg up to about 100 mg / kg, or more, depending on the factors described above. In certain embodiments, dosages may range from 0.1 μg / kg up to about 100 mg / kg, or from 1 μg / kg up to about 100 mg / kg, or from 5 μg / kg up to about 100 mg / kg. Another exemplary, non-limiting range for a therapeutically or prophylactically effective amount of an antibody or antibody portion is 0.1 to 20 mg / kg, more preferably 1 to 10 mg / kg. It should be noted that dosage values may vary depending on the type and severity of the disease, disorder, or condition to be alleviated. Furthermore, for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and it should be understood that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or embodiments of the claimed compositions.

[0147] Fixed doses and fixed dosing regimens, such as by subcutaneous administration, are also contemplated herein. Exemplary fixed doses contemplated for use herein can be selected from 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, 2000 mg, etc. These fixed doses can be used either as a loading dose or as subsequent maintenance doses. Thus, exemplary dosing regimens contemplated for use herein include, among others, a 1200 mg loading dose and a 600 mg maintenance dose, a 1200 mg loading dose and a 300 mg maintenance dose, a 900 mg loading dose and a 600 mg maintenance dose, a 900 mg loading dose and a 300 mg maintenance dose, a 800 mg loading dose and a 400 mg maintenance dose, a 800 mg loading dose and a 300 mg maintenance dose, a 800 mg loading dose and a 200 mg maintenance dose, a 200 mg loading dose and a 400 mg maintenance dose, a 200 mg loading dose and a 3 ... 00mg maintenance dose, 700mg loading dose and 300mg maintenance dose, 600mg loading dose and 300mg maintenance dose, 600mg loading dose and 250mg maintenance dose, 600mg loading dose and 200mg maintenance dose, 600mg loading dose and 150mg maintenance dose, 600mg loading dose and 100mg maintenance dose, 300mg loading dose and 150mg maintenance dose, 300mg loading dose and 100mg maintenance dose, etc.

[0148] In another aspect, the present application provides methods for detecting the presence of IL1RAP in an in vitro sample (e.g., a biological sample such as serum, plasma, tissue, biopsy, etc.). The subject methods can be used to diagnose a disease, disorder, or condition. The methods include (i) contacting the sample or a control sample with an anti-IL1RAP antibody or fragment thereof described herein, and (ii) detecting the formation of a complex between the anti-IL1RAP antibody or fragment thereof and the sample or control sample, wherein a statistically significant change in the formation of the complex in the sample relative to the control sample indicates the presence of IL1RAP in the sample.

[0149] Given their ability to bind to human IL1RAP, anti-human IL1RAP antibodies or portions thereof (as well as ADCs thereof) can be used to detect human IL1RAP (e.g., in biological samples such as serum or plasma) using conventional immunoassays, such as enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), or tissue immunohistochemistry. In one embodiment, the present disclosure provides a method for detecting human IL1RAP in a biological sample, comprising contacting the biological sample with an antibody or antibody portion and detecting either the antibody (or antibody portion) bound to human IL1RAP or the unbound antibody (or antibody portion), thereby detecting human IL1RAP in the biological sample. The antibody is directly or indirectly labeled with a detectable substance to facilitate detection of the bound or unbound antibody. Suitable detection substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase. Examples of suitable prosthetic groups include streptavidin / biotin and avidin / biotin. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin. An example of a luminescent material is luminol. Examples of suitable radioactive materials include 3 H, 14 C.35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, or 153 There is Sm.

[0150] Instead of labeling the antibody, human IL1RAP can be measured in body fluids by a competitive immunoassay utilizing a rhIL1RAP standard labeled with a detectable substance and unlabeled anti-human IL1RAP. In this assay, a biological sample, labeled rhIL1RAP standard, and anti-human IL1RAP antibody are combined, and the amount of labeled rhIL1RAP standard bound to the unlabeled antibody is determined. The amount of human IL1RAP in the biological sample is inversely proportional to the amount of labeled rhIL1RAP standard bound to the anti-IL1RAP. Similarly, human IL1RAP can also be measured in body fluids by a competitive immunoassay utilizing a rhIL1RAP standard labeled with a detectable substance and unlabeled anti-human IL1RAP antibody.

[0151] In yet another aspect, the present application provides a method of detecting the presence of IL1RAP in vivo (e.g., in vivo imaging in a subject). The subject method can be used to diagnose a disorder, e.g., an IL1RAP-associated disorder. The method includes (i) administering an anti-IL1RAP antibody or fragment thereof described herein to a subject or a control subject under conditions that allow binding of the antibody or fragment to IL1RAP, and (ii) detecting formation of a complex between the antibody or fragment and IL1RAP, wherein a statistically significant change in complex formation in the subject relative to the control subject indicates the presence of IL1RAP.

[0152] V. Pharmaceutical Compositions The present disclosure also provides pharmaceutical compositions comprising an antibody or antigen-binding portion thereof, or an ADC, and a pharmaceutically acceptable carrier. Pharmaceutical compositions comprising an antibody or ADC are intended for use in, but not limited to, diagnosing, detecting, or monitoring a disorder, preventing, treating, managing, or ameliorating a disorder or one or more symptoms thereof, and / or in research. In certain embodiments, the composition comprises one or more antibodies. In another embodiment, the pharmaceutical composition comprises one or more antibodies or ADCs and one or more prophylactic or therapeutic agents other than antibodies or ADCs for treating an inflammatory, autoimmune, or atopic disease, disorder, or condition, or a disease, disorder, or condition in which IL1RAP activity is deleterious. Preferably, the prophylactic or therapeutic agents are known, have been used, or are currently used to prevent, treat, manage, or ameliorate a disorder or one or more symptoms thereof. In accordance with these embodiments, the composition may further comprise a carrier, diluent, or excipient.

[0153] Antibodies and antibody portions or ADCs can be incorporated into pharmaceutical compositions suitable for administration to a subject. Typically, pharmaceutical compositions comprise an antibody or antibody portion and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, as well as those described in US Pat. No. 8,829,165 and US Pat. No. 8,859,741 (which are incorporated by reference in their entireties for all purposes). Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate-buffered saline, dextrose, glycerol, ethanol, and the like, as well as combinations thereof. In many cases, it will be preferable to include an isotonic agent, for example, a sugar, a polyalcohol such as mannitol or sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable carriers may contain minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives, or buffers, which enhance the shelf life or effectiveness of the antibody or antibody portion or ADC. The bioavailability of an antibody or antibody portion can be increased by including an excipient such as hyaluronidase in the carrier or composition. The viscosity of a composition, carrier, antibody, or antibody portion can be increased by including an excipient such as caffeine.

[0154] Various delivery systems are known and can be used to administer one or more antibodies or ADCs, or a combination of one or more antibodies and prophylactic or therapeutic agents, useful for preventing, managing, treating, or ameliorating a disorder or one or more symptoms thereof, including, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the antibody or antibody fragment, receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)), and construction of a nucleic acid as part of a retrovirus or other vector. Methods of administering prophylactic or therapeutic agents include, but are not limited to, parenteral administration (e.g., intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous), epidural administration, intratumoral administration, and mucosal administration (e.g., intranasal and oral routes). In addition, pulmonary administration can be employed, for example, by use of an inhaler or nebulizer and formulation with an aerosolizing agent. See, e.g., U.S. Patent No. 6,019,968 and WO 99 / 66903, each of which is incorporated herein by reference in its entirety. In one embodiment, the antibody, combination therapy, or composition is administered using Alkermes AIR® pulmonary drug delivery technology (Alkermes, Inc., Cambridge, Mass.). In specific embodiments, the prophylactic or therapeutic agent is administered intramuscularly, intravenously, intratumorally, orally, intranasally, pulmonary, or subcutaneously. The prophylactic or therapeutic agent is administered by any convenient route, e.g., by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral, rectal, and intestinal mucosa), and may be administered in conjunction with other biologically active agents. Administration can be systemic or local.

[0155] In certain embodiments, it may be desirable to administer prophylactic or therapeutic agents locally to the area in need of treatment. This may be accomplished, for example, but not limited to, by local infusion, injection, or implant, where the implant is of a porous or non-porous material, including membranes and matrices, such as sialurastic membranes, polymers, fibrous matrices (e.g., Tissuel®), or collagen matrices. In one embodiment, an effective amount of one or more antibody antagonists is locally administered to the affected area of a subject to prevent, treat, manage, and / or ameliorate a disorder or a symptom thereof. In another embodiment, an effective amount of one or more antibodies is locally administered to the affected area in combination with an effective amount of one or more therapies other than the antibody (e.g., one or more prophylactic or therapeutic agents) to prevent, treat, manage, and / or ameliorate a disorder or one or more symptoms thereof.

[0156] A pharmaceutical composition is formulated to be compatible with its intended route of administration. Examples of routes of administration include, but are not limited to, parenteral administration, e.g., intravenous administration, intradermal administration, subcutaneous administration, oral administration, intranasal administration (e.g., inhalation), transdermal administration (e.g., topical administration), transmucosal administration, and rectal administration. In certain embodiments, the composition is formulated according to routine procedures as a pharmaceutical composition adapted for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to humans. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. If necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the injection site.

[0157] When the method of the present disclosure involves oral administration, the composition can be orally formulated in the form of tablets, capsules, cachets, gelcaps, solutions, suspensions, etc. Tablets or capsules can be prepared by conventional means with pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose), fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate), lubricants (e.g., magnesium stearate, talc, or silica), disintegrants (e.g., potato starch or sodium starch glycolate), or wetting agents (e.g., sodium lauryl sulfate). Tablets may be coated by methods well known in the art. Liquid preparations for oral administration may take the form of, but are not limited to, solutions, syrups, or suspensions, and may also be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats), emulsifying agents (e.g., lecithin or acacia), non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, or fractionated vegetable oils), and preservatives (e.g., methyl or propyl-p-hydroxybenzoate esters, or sorbic acid). The preparations may also contain buffer salts, flavoring agents, coloring agents, and sweetening agents, as appropriate. Preparations for oral administration may be suitably formulated for slow, controlled, or sustained release of the prophylactic or therapeutic agent(s).

[0158] The method may involve administering a composition formulated for parenteral administration by injection (e.g., bolus injection or continuous infusion). Formulations for injection may be presented in unit dosage form (e.g., in ampoules or in multi-dose containers) with an added preservative. The composition may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle (e.g., sterile, pyrogen-free water) before use.

[0159] Generally, the components of the composition are supplied separately or mixed in unit dosage form in a sealed container such as an ampoule or a small bag that indicates the amount of active agent, for example, as dry lyophilized powder or water-free concentrate.When the administration method is infusion, the composition can be prepared using an infusion bottle that contains sterile pharmaceutical grade water or saline.When the administration method is injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed before administration.

[0160] In particular, the present disclosure also provides that one or more of the prophylactic or therapeutic agents or pharmaceutical compositions are packaged in a hermetically sealed container, such as an ampoule or sachet, indicating the quantity of agent. In one embodiment, one or more of the prophylactic or therapeutic agents or pharmaceutical compositions are supplied as a dry, sterilized, lyophilized powder or water-free concentrate in a hermetically sealed container, which can be reconstituted (e.g., with water or saline) to the appropriate concentration for administration to a subject. Antibodies and antibody portions or ADCs can be administered by a variety of methods known in the art; however, for many therapeutic applications, the preferred route / mode of administration is subcutaneous injection, intravenous injection, or infusion. As will be appreciated by those skilled in the art, the route and / or mode of administration will vary depending on the desired results. In certain embodiments, the active compound may be prepared with a carrier that will protect the compound against rapid release, such as a controlled-release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art (see, e.g., Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978).

[0161] It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods of the present invention described herein will be obvious and may be made using suitable equivalents without departing from the scope of the invention or the embodiments disclosed herein. Having set forth a detailed description of the invention above, it will be more clearly understood by reference to the following examples, which are provided for illustrative purposes only and are not intended to be limiting. [Example]

[0162] Example 1. IL1RAP is expressed in leukemia cell lines To examine the expression of IL1RAP protein in leukemia cell lines, the following experiment was carried out. method Tissue culture and cell lines Human leukemia cell lines EOL1, Monomac6, OCI / AML1, KG-1, and Karpas299 were obtained from DSMZ. Cells were maintained in RPMI-1640 medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS) (Sigma). Flow cytometry Staining for flow cytometry was performed in cold 1x PBS containing 0.5% BSA. Primary antibodies (1 μg / ml) were incubated with live cells for 30 minutes on ice. After a brief wash, cells were incubated with Alexa Fluro® 488-conjugated anti-mouse IgG secondary antibody at 1:1000 (#4408, Cell Signaling Technology). Data were acquired using MACSQuant® Flow Cytometers (Miltenyi Biotec) and analyzed with FlowJo® software. result Using a mouse monoclonal anti-IL1RAP antibody (MAB676, R&D System), we examined the surface expression of IL1RAP in a panel of leukemia cell lines. Flow cytometry analysis was performed on the acute myeloid leukemia cell lines EOL1, Monomac6, OCI / AML1, and KG-1, as well as the T-cell leukemia cell line Karpas299. Abundant surface expression of IL1RAP was observed in all cell lines tested (see Figure 1).

[0163] Example 2. Generation of human monoclonal antibodies against the IL1RAP extracellular domain To generate a fully human antibody (IL1RAP-ECD) against the extracellular domain of IL1RAP (SEQ ID NO: 260), the following experiment was carried out.

[0164] method

[0165] Immunization in humanized mice

[0166] Monoclonal antibodies were obtained by immunization with 293T cells expressing either recombinant human IL1RAP-ECD or full-length human IL1RAP with a C-terminal MYC-DDK tag. Genetically engineered mice were engineered to produce human immunoglobulins with variable regions. Mice received five doses of either recombinant protein or cells via intraperitoneal injection (IP) and were then allowed to rest for one month. Mice were then boosted 4 and 2 days before fusion with rabbit spleen cells expressing full-length IL1RAP or recombinant IL1RAP extracellular domain (ECD). Human IL1RAP-ECD recombinant protein was expressed in EBNA293 cells and purified. For each hybridoma, each variable domain was cloned by RT-PCR into an expression vector providing the appropriate constant region. Four plasmid isolates from each cloning were subjected to Sanger sequencing. After analysis, unique recombinant heavy chains were paired with unique recombinant light chains. These plasmid pairs were transfected into CHO cells in 24-well plates. After 8-12 days, the conditioned medium from each pair was screened for binding to IL1RAP using FLOW™ or Octet™.

[0167] Recombinant IL1RAP cloning

[0168] Human, rat, and mouse IL1RAP cDNAs were purchased from Origene (RC211970, RR213032, MR223729, Rockville, MD). The encoded proteins are 100% identical to GenBank's IL1RAP_HUMAN. Macaca fascicularis IL1RAP cDNA was synthesized from Gen9 (CST-35853, Cambridge, MA).

[0169] The extracellular domains of human, Macaca fascicularis, rat, and mouse were cloned by PCR, and the synthetic gene was based on sequences in GenBank (see Table 1).

[0170] All DNA sequences were cloned into appropriate CMV-based expression vectors with non-native signal peptides and C-terminal histidine tags for purification.

[0171] [Table 1] Using a retroviral MSCV construct, full-length human IL1RAP protein was expressed on the surface of HEK-293T, CHO, and rabbit splenocytes (see Table 2). Mouse, rat, and Macaca fascicularis IL1RAP were also expressed on the surface of HEK-293T.

[0172] [Table 2] A series of plasmid constructs designed to secrete soluble IL1RAP extracellular domains were constructed from the full-length plasmid. Each construct in Table 3 was cloned as a fusion protein with an N-terminal maltose-binding protein (MBP) and a C-terminal eight-histidine (8xHis) tag. Both human and mouse constructs were generated.

[0173] [Table 3]

[0174] Cloning of VH and VL sequences from hybridomas To determine the CDR sequences, total RNA was isolated from hybridoma cells using the RNeasy® kit (Qiagen, Hilden, Germany). First- and second-strand cDNA synthesis was performed using the OneTaq® one-step RT-PCR kit (New England BioLabs, Ipswich, MA). Several primers were used, including those listed in Table 4 of U.S. Patent 11,248,054 and SEQ ID NOs: 260-285 of U.S. Patent 11,248,054 (which are incorporated herein by reference in their entirety for all purposes). PCR products were separated by agarose gel electrophoresis, and fragments were excised and purified with a QIAquick® gel extraction kit (Qiagen, Hilden, Germany). Fragments were directly cloned into expression vectors using Golden Gate cloning technology with BspQI (New England BioLabs, Ipswich, MA). Four colonies from each reaction were scaled up and subjected to miniprep-scale plasmid purification using the SequeMid® DNA Purification Kit (Aline Biosciences, Woburn, Mass.).

[0175] A system for mid-scale antibody production or transient expression of recombinant proteins.

[0176] IL1RAP recombinant protein and anti-IL1RAP antibody were expressed in Chinese hamster ovary (CHO) cells in one shake flask (100 mL working volume) using the recommended transfection and media components of the ExpiCHO™ system (Invitrogen, Carlsbad, CA). Cell culture supernatants were harvested 14 days post-transfection, centrifuged, and filtered (0.22 μm).

[0177] Antibody and protein purification Conditioned media from CHO cell cultures was clarified, filtered, and then loaded onto an ÅKTA pure™ system equipped with a 5 mL MabSelect SuRe® column (GE Healthcare) for purification. The antibody was eluted with 100 mM glycine, pH 3.5, neutralized with 1 M Tris-Cl, pH 8.5, and dialyzed against PBS.

[0178] The recombinant target protein was purified from the conditioned medium by Ni-NTA chromatography. The His-tagged protein was eluted and dialyzed against PBS.

[0179] Recombinant antibody analysis Concentration: The concentration of recombinant antibodies was determined on a Fortebio Octet Red™ (Pall ForteBio, Fremont, CA) instrument using a protein A chip and a human IgG1 antibody for the standard curve.

[0180] Purity test by SDS-PAGE: Purity tests were performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of reduced and non-reduced samples. Samples (10 μg) were mixed with loading buffer (+ / - β-mercaptoethanol), heated, and electrophoresed on a 4-20% gel (Invitrogen, Carlsbad, CA). Bands were visualized with Coomassie InstantBlue™ (Expedeon, San Diego, CA) staining.

[0181] Endotoxin Purity Testing: Endotoxin concentrations were measured by Limulus Amoebocyte Lysate (LAL) kinetic turbidimetric assay using the Endosafe-PTS™ system (Charles River Laboratories, Wilmington, MA).

[0182] Purity testing by HPLC-SEC: Samples were screened for antibody aggregation and other forms using a 1260 Infinity System™ (Agilent, Santa Clara, CA) equipped with a TSKgel UltraSW Aggregate Guard™ column and an HPLC column (Tosoh Bioscience). Samples and standards were detected by absorbance at 280 nm. Molar masses of sample components were determined by comparison with a standard curve.

[0183] Affinity: The affinity of antibodies to various recombinant IL1RAP proteins was measured using an Octet Red™ instrument. After loading the reagents into a 96-well plate, the Octet Red™ instrument with a Protein A-labeled biosensor was programmed as follows: 30 seconds for baseline #1, 120 seconds for antibody immobilization, 30 seconds for baseline #2, 300 seconds for antibody binding to recombinant IL1RAP, and 300–600 seconds for dissociation of recombinant IL1RAP from the antibody.

[0184] Binding competition binning: Binding competition between different antibodies was determined using a real-time interferometry assay using Octet Red™ with a Protein A-labeled biosensor. To assess whether two antibodies compete for binding to recombinant IL1RAP protein, the assay was performed as follows: Protein A biosensors were first immersed in wells containing 10 μg / mL of a monoclonal antibody for 5 minutes. After the capture step, the biosensors were briefly immersed in buffer (15 seconds), and then unoccupied sites on the biosensors were saturated by immersion in wells containing 100 μg / mL of an irrelevant monoclonal antibody for 5 minutes. Octet™ biosensors were briefly immersed in buffer (15 seconds) before being immersed in wells containing recombinant IL1RAP for 1 minute. The biosensors were briefly immersed in buffer (15 seconds) before being immersed in wells containing a second recombinant antibody for 1 minute.

[0185] In the control case where the second antibody was the same as the first antibody, no increase in signal was observed due to the lack of additional binding to the recombinant target.

[0186] In the control case where buffer was used in place of the first antibody, the recombinant target did not bind to the unquenched antibody on the biosensor, and the second antibody also did not bind to the biosensor.

[0187] If the signal increased with the second antibody, the two antibodies were determined to not compete.

[0188] If the signal was not enhanced with the second antibody, the two antibodies were determined to compete for binding.

[0189] Immunofluorescence (IF)-based high content screening (HCS) High-content immunofluorescence was used to identify wells containing immunoglobulins that preferentially bound to cells expressing IL1RAP. Briefly, CHO cells and CHO-hIL1RAP cells seeded 24 h prior to the assay were incubated with hybridoma supernatant diluted 2-fold in DMEM + 10% fetal bovine serum (FBS) for 60 min at 37°C. After incubation, cells were fixed with 4% formaldehyde, washed with PBS, permeabilized with 0.3% Triton-X-100, and labeled with anti-rat Alexa488 (hybridoma stage) or anti-human Alexa488 secondary antibodies (with recombinant IL1RAP antibody) for 1 h at room temperature. Unbound secondary antibodies were removed with PBS washes, and cells were stained with DNA dyes (propidium iodide and Hoechst 33342).

[0190] Potential hits were initially identified by low-resolution, high-throughput screening using a TTP Labtech Acumen eX3™ (TTP Labtech, Cambridge, MA) to quantify the difference in fluorescence for each sample in positive and negative cell lines. These hits were then validated and characterized for subcellular localization using a Thermo ArrayScan VTi™ (Thermo Fisher Scientific, Waltham, MA) to obtain high-resolution images of both cell lines.

[0191] Flow cytometry Staining for flow cytometry was performed in cold 1x PBS containing 0.5% BSA. Primary antibodies (1 μg / ml) were incubated with live cells for 30 minutes on ice. After a brief wash, cells were incubated with Alexa Fluro® 488-conjugated anti-human IgG secondary antibody (709-546-149, Jackson ImmunoResearch) at a dilution of 1:1000. Data were acquired using MACSQuant® Flow Cytometers (Miltenyi Biotec) and analyzed with FlowJo software.

[0192] Correction of sequence defects There are many amino acid sequences that are predictive of poor performance in clinical-scale production and stability, such as non-consensus cysteine residues (Cys), non-consensus N-linked glycosylation sites (Asn-xxx-Ser / Thr), and acid-sensitive sequences (Asp-Pro). Some antibodies obtained from hybridoma cloning contain one or more of these sequences and yet have the desired biological properties.

[0193] Some antibody sequences with such sequence disorders were engineered to remove the disorder in order to preserve or improve binding properties. Antibodies with non-consensus Cys residues were mutated by replacing Cys with the germline sequence (if Cys is in the framework), Ser, or Ala. Constructs of this type were generated, expressed in CHO cells, and then functionally tested.

[0194] In one example, antibody 5D12_18A4, with a dissociation constant (KD) of 19 nM, contains a non-consensus Cys sequence in the VH sequence. This heavy chain sequence was engineered to contain a Cys108Tyr mutation. This new heavy chain plasmid was paired with the original light chain plasmid and transfected into CHO cells. Antibody expression and affinity for human IL1RAP were screened. The 5D12-C108Y antibody was expressed at the same level as the 5D12_18A4 antibody and had a KD of 13 nM.

[0195] In the second case, antibody 10C8_15A1, with a dissociation constant (KD) of 30 nM, contains a non-consensus Cys sequence in the VH sequence. This heavy chain sequence was engineered to contain a Cys43Ala mutation. This new heavy chain plasmid was paired with the original light chain plasmid and transfected into CHO cells. Antibody expression and affinity for human IL1RAP were screened. The 10C8_C43A antibody was expressed at the same level as the 10C8_15A1 antibody, with a KD of 13 nM.

[0196] Some antibody sequences with non-consensus N-linked glycosylation sites are modified at either the Asn or Ser / Thr sites. When possible, the Asn or Ser / Thr codons can be mutated to the germline sequence. Additionally, replacing Asn with Gln or a similar amino acid, and Ser or Thr with a similar or smaller amino acid, may also be successful.

[0197] In one example, antibody 32C12_21A4, with a dissociation constant (KD) value of 1 nM, contains a non-consensus N-linked glycosylation site in CDR1 of the VL sequence. The light chain was engineered to contain an Asn26Ser mutation. This new light chain plasmid was paired with the original heavy chain sequence and transfected into CHO cells. Antibody expression and affinity for human IL1RAP were screened. The 32C12-N26S antibody showed 40% higher expression than the 32C12 antibody. The KD value of the 32C12-N26S antibody is 19 nM.

[0198] The complete amino acid sequences of the heavy and light chains of the 36 antibodies are shown in Table 5 below.

[0199] [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6] [Table 4-7] [Table 4-8] [Table 4-9] [Table 4-10] [Table 4-11] [Table 4-12]

[0200] The complete nucleic acid sequences encoding the heavy and light chains from these 36 antibodies are set forth in Table 6 of U.S. Patent 11,248,054 and as SEQ ID NOs: 191 to 259 of U.S. Patent 11,248,054, and are incorporated herein by reference in their entirety for all purposes.

[0201] The affinities of the 36 antibodies were calculated by Octet™ using the huIL1RAP ECD recombinant protein and ranged from 0.2 nM to 127 nM (see Table 7 below).

[0202] [Table 5]

[0203] result Fully human antibodies against IL1RAP were generated using the hybridoma method. Briefly, transgenic mice were immunized with recombinant human IL1RAP-ECD or 293T cells overexpressing IL1RAP and boosted with rabbit splenocytes expressing full-length human IL1RAP or the ECD of the IL1RAP recombinant protein. Splenocytes were fused with the mouse myeloma cell line X63-Ag8.653. Clones from the transgenic mice were identified by immunofluorescence (IF)-based high-content screening (HCS) using CHO cells overexpressing hIL1RAP and parental CHO cells that do not express IL1RAP.

[0204] Over 1,000 hits were identified that strongly bound to CHO-human IL1RAP but not to parental CHO cells. 68 clones were selected for molecular cloning. Based on unique CDR3 sequences from the heavy chain variable domain, 35 antibodies from 13 families were confirmed by binding in the IL1RAP-positive cell lines EOL1 and Karpas299 by flow cytometry (MACSQuant®, www.miltenybiotec.com) (see Table 8). The binding characteristics of a representative antibody clone, 44E5_15C5, are shown in Figure 2.

[0205] [Table 6]

[0206] Selected antibodies were evaluated in pairs for their ability to simultaneously bind recombinant human IL1RAP. Analysis was performed using an Octet™ equipped with a Protein A biosensor. A total of five distinct bins were determined as follows. Some of the bins overlapped with each other, and an antibody may fall into multiple bins. (1) The following five antibodies competed with each other for binding: 5D12_18A4, 24G3_17C5, 34C11_21B2, 38E10_21C3, 39G1_21C4. (2) The following six antibodies competed with each other: 10c8_15a1, 16h2_17d2, 32c12_21a4, 37e10_15b5, 44e5_15c5, 5g8_18a1. (3) Two antibodies competed with each other in unique configurations: 5G8_18A1, 36A10_21B6. (4) Two antibodies competed with each other in unique configurations: 36A10_21B6, 37D11_21C2. (5) Two antibodies competed with each other in unique configurations: 37D11_21C2, 12F3_17C2. A diagram depicting the placement of these competing bins is shown in Figure 3.

[0207] Example 3 Binding of anti-IL1RAP monoclonal antibodies to IL1RAP orthologs

[0208] To investigate the binding of anti-IL1RAP human monoclonal antibody to IL1RAP, experiments were carried out in various species using the following method.

[0209] method Tissue culture and cell lines The 293T cell line was purchased from the American Type Culture Collection (ATCC). 293T cells expressing human, macaca fascicularis, rat, and mouse IL1RAP were maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS) (Sigma) in the presence of 2 μg / ml puromycin (Invitrogen).

[0210] Flow cytometry Staining for flow cytometry was performed in cold 1x PBS containing 0.5% BSA. Primary antibodies (1µg / ml) were incubated with live cells for 30 minutes on ice. After a brief wash, cells were incubated with Alexa Fluro® 488-conjugated anti-human IgG secondary antibody (709-546-149, Jackson ImmunoResearch) at 1:1000. Data were acquired using MACSQuant® Flow Cytometers (Miltenyi Biotec) and analyzed with FlowJo software.

[0211] result To evaluate the binding of anti-IL1RAP human monoclonal antibodies to IL1RAP from different species, we constructed 293 cells overexpressing human, macaca fascicularis, rat, and mouse IL1RAP. All antibodies bound to human and macaca fascicularis IL1RAP (Figures 4A-4B), whereas 24G3_17C5 and 34C11_21B2 bound strongly to 293 cells expressing rat IL1RAP, and 24G3_17C5, 5D12_18A4, and 39G1_21C4 bound weakly to 293 cells expressing mouse IL1RAP (Figures 4C-4D).

[0212] Example 4. Internalization of anti-IL1RAP antibodies in AML cell lines Experiments were performed to characterize the internalization of anti-IL1RAP antibodies in EOL-1 cells. The following methods were used.

[0213] method Tissue culture and cell lines The human leukemia cell line EOL1 was obtained from DSMZ and maintained in RPMI-1640 medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS) (Sigma).

[0214] Internalization assay Live EOL1 cells were incubated with 44E5_15C5 antibody for 30 minutes at 37°C. After cytospinning, the cells were fixed with 4% PFA, permeabilized with 100% methanol, and stained with LAMP1 antibody (#9091, Cell Signaling Technology, Inc.).

[0215] result Live EOL1 cells were incubated with 44E5_15C5 for 0.5 hours at 37°C. The cells were then fixed, permeabilized, and co-stained with LAMP1 antibody. 44E5_15C5 co-localized with lysozyme labeled with LAMP1 antibody (see Figure 5).

[0216] Example 5. Blockade of IL1 signaling by anti-IL1RAP antibody To characterize the ability of anti-IL1RAP antibodies to block IL1 signaling, the following experiments were performed.

[0217] method IL-1 signaling reporter cell assay HEK-Blue IL-1β cells (Invivogen, CA) were harvested and plated in technical replicates in 96-well plates at a density of 50,000 cells per well. IL1RAP antibody or the corresponding human IgG1 control antibody was added to the wells at concentrations ranging from 1 to 10 μg / ml. After 30 minutes of incubation with the antibody, IL-1β was added to a final concentration of 0.5 ng / ml, and the plates were incubated overnight. To determine whether any antibody could induce IL-1R activation in the absence of IL-1β, samples were incubated with 10 μg / mL of antibody without the addition of ligand. The following day, substrate was added to the supernatant, and the absorbance of the samples was measured at 620 nm.

[0218] result Because IL1RAP is essential for IL1 signaling, we examined the ability of the IL1RAP antibodies described herein to inhibit IL-1 signaling. Although the antibodies exhibited varying degrees of inhibitory effect in the IL-1 reporter assay, the IL1RAP antibodies 37E10_15B5, 44E5_15C5, 16H2_17D2, 5G8_18A1, and 36A10_21B6 potently inhibited IL1R1 signaling in a dose-dependent manner (Figure 6A).

[0219] None of the IL1RAP antibodies tested affected IL1R1 signaling in the absence of IL-1, thus excluding an agonistic function on IL1-induced signaling.

[0220] In addition, serial dilutions of the IL1RAP antibody were performed to measure the EC values against IL-1β and IL-1α. 50 Many of these antibodies, including 37E10_15B5 and 44E5_15C5, had subnanomolar EC 50 can block IL-1β and IL-1α signaling, whereas 37D11_21C2 and 39G1_21C4 do not inhibit IL-1 signaling.

[0221] Example 6. Blockade of IL-33 signaling by anti-IL1RAP Ab

[0222] Experiments were conducted to characterize the ability of anti-IL1RAP antibodies to block IL-33 signaling using the following methods.

[0223] method IL-33 signaling reporter cell assay HEK-Blue IL-33 cells (Invivogen, CA) were harvested and plated in technical duplicates at a density of 50,000 cells per well in 96-well plates. Antibodies or the corresponding human IgG1 control antibody were added to the wells at concentrations ranging from 1 to 10 μg / ml. After incubating the cells with the antibodies for 30 minutes, IL-33 was added to a final concentration of 0.5 ng / ml, and the plates were incubated overnight. To determine whether any of the antibodies could induce ST2 activation in the absence of IL-33, samples were incubated with 10 μg / mL of antibody without the addition of ligand. The following day, substrate was added to the supernatant, and the absorbance of the samples was measured at 620 nm.

[0224] result Because IL1RAP plays a critical role in IL-33 signaling, we examined the ability of the developed antibodies to inhibit IL-33 signaling. The antibodies showed varying degrees of inhibitory effects in the IL-33 reporter assay, but 37E10_15B5, 44E5_15C5, and 36A10_21B6 were able to partially inhibit IL-33 signaling in a dose-dependent manner (see Figures 7A and 7B). None of the antibodies affected IL-33 signaling in the absence of IL-33, excluding their antagonistic role in IL-33-induced signaling (data not shown). On the other hand, 37D11_21C2 and 39G1_21C4 did not inhibit IL-33 signaling.

[0225] Example 7. Efficacy of anti-IL1RAP Ab in atopic dermatitis model mice

[0226] To clarify the efficacy of anti-IL1RAP antibodies in a mouse model of atopic dermatitis, we conducted an experiment using the following method.

[0227] method Atopic dermatitis model in oxazolone / HDM mice

[0228] Eight-week-old Balb / c mice were used. To induce atopic dermatitis, mice received either no model induction (n = 3; Group 1), an injection of olive oil:acetone vehicle (1:4; n = 3; Group 2), or 0.8% oxazolone in olive oil:acetone vehicle plus house dust mite (HDM) (n = 10 each; Groups 3 and 4). Oxazolone plus HDM was administered on days 0 (D0), 5, 8, 12, 15, and 17.

[0229] Mice in the oxazolone + HDM group were intraperitoneally injected with either the anti-IL1RAP antibody of the present invention (Group 4; n=10) or an isotype control antibody (Group 3; n=10) at a dose of 20 mg / kg on days 3, 6, 10, 13, and 17. Mice in the no-model and vehicle-only groups (Groups 1 and 2) were intraperitoneally injected with saline. The anti-IL1RAP antibody 19D2 used in this example is a surrogate antibody used in mouse studies that corresponds to mouse IgG1 (similar to human IgG4) directed against mouse IL-1RAP.

[0230] The body weight and skin condition of the mice were monitored twice weekly. Pruritus was assessed on day 17 by scratching the affected area and quantified as the number of scratches per 30 minutes. On day 18, the animals were sacrificed, and blood, skin, and spleen samples were collected. To analyze the levels of soluble inflammatory / atopic mediators, skin samples were homogenized in the presence of protease inhibitors and subjected to ELISA for the mediators of interest. To analyze the levels of inflammatory cells, skin samples were minced, digested with collagenase D, and subjected to cell sorting to obtain live CD45+ cells, which were then stained with fluorescently labeled antibodies. The experimental design is shown in Figure 8.

[0231] result As shown in Figure 9, treatment with anti-IL1RAP antibody significantly reduced pruritus, a key symptom of human atopic dermatitis, by D17 compared with treatment with an isotype control antibody. In the oxazolone + HDM model, mice exhibited increased skin thickness (another key symptom of human atopic dermatitis), increased spleen size (splenomegaly), and suppressed weight gain. Here, by D18, treatment with anti-IL1RAP antibody prevented or reduced skin thickening and spleen enlargement, and the skin and spleens of treated mice were thinner and smaller, respectively, than those of mice treated with an isotype control. Furthermore, treatment with anti-IL1RAP antibody significantly reduced the inhibition of weight loss, and treated mice gained more weight than mice treated with an isotype control. These findings are shown in Figures 10A, 10B, and 10C. Thus, treatment with anti-IL1RAP antibody significantly reduced the symptoms and pathology associated with human atopic dermatitis. Furthermore, as determined by ELISA and shown in Figure 11, treatment with anti-IL1RAP antibody significantly reduced the levels of the inflammatory / atopic mediators eotaxin, lipocalin-2, TARC / CCL17, TSLP, and IL-6 in skin at D18 compared to treatment with the isotype control. Furthermore, treatment with anti-IL1RAP antibody significantly reduced the levels of neutrophils, macrophages, and eosinophils, i.e., immune cells that mediate inflammatory / atopic responses, in skin at D18 compared to the isotype control, as shown in Figure 12. These findings provide molecular and cellular evidence supporting the efficacy of the anti-IL1RAP antibody of the present invention in atopic dermatitis.

[0232] BFB759 in vitro testing Multiple in vitro studies were conducted to investigate whether BFB759 could be successfully combined with various approved and effective biologics and small molecule therapeutics for inflammatory diseases such as atopic dermatitis and asthma. These studies confirmed that therapeutic agents targeting IL-4Ra, IL-13, TNFα, and Janus kinases exhibited additive or synergistic effects when combined with BFB759. Furthermore, the widely used corticosteroid dexamethasone also exhibited additive or synergistic effects when combined with BFB759. These studies suggest that combination therapy with multiple anti-inflammatory agents may be useful when targeting IL-1R3 in inflammatory, autoimmune, and autoinflammatory diseases.

[0233] Example 8. Inhibitory effect of BFB759 on TARC production in HDM-stimulated human PBMCs To characterize the inhibition of TARC production in HDM-stimulated human PBMCs using BFB759, the following method was used.

[0234] method Materials included human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors, frozen in liquid nitrogen, and either thawed or freshly prepared before analysis. HDM (house dust mite extract) was from the Greer lab and purchased through Fisher Scientific (NC9756554, Lot #390844). Human IL-4 (204-IL-010) was purchased from R&D Systems. Human IL-13 (200-13) was purchased from Peprotech. The JAK inhibitors (JAKi) abrocitinib (HY-107429) and upadacitinib (HY-19569) were purchased from MedChemExpress. Dexamethasone (D4902) was purchased from Sigma-Aldrich. Dupilumab (HY-P9926) was purchased from MedChemExpress. Tralokinumab was manufactured by Bluefin Biomedicine according to the published protein sequence, and the TARC ELISA kit (DuoSet Cat#DY364, Quantikine Cat#SDN00) was purchased from R&D Systems.

[0235] Blood was collected from healthy volunteers in heparinized tubes. PBMCs were isolated by centrifugation of blood over a Ficoll-Paque Plus density gradient using Leucosep tubes. 0.2 million PBMCs in 100 μl of medium (RPMI 1640, 10% heat-inactivated FBS) were added to each well of a 96-well cell culture plate. BFB759 at various concentrations in 50 μl of medium was added to the wells. The cells were incubated in an incubator for 45 minutes. Then, 50 μl of HDM solution (final concentration 10 μg / ml) was added to the wells. After 7 days of incubation, the supernatants were transferred to a new 96-well plate and either used immediately for TARC ELISA or frozen in a -80°C freezer for later use in ELISA. For BFB759 combination testing with other drugs, single drugs or drug combinations were added to PBMCs and incubated for 45 minutes, after which HDM solution, or HDM with IL-4 or IL-13 solution, was added to the wells. Analytical samples were cultured for 7 days and supernatants were collected for TARC ELISA.

[0236] result Thymus and activation-regulated chemokine (TARC) plays an important role in allergic diseases such as atopic dermatitis and asthma. High serum TARC levels are observed in patients with atopic dermatitis, and these levels are closely related to disease activity. Measurement of serum TARC levels has already been clinically applied as a useful marker for atopic dermatitis disease activity. In this study, PBMCs from different donors were stimulated with HDM and treated with BFB759 at a wide range of concentrations from 0.1 μg to 150 μg / ml. Depending on the donor, BFB759 can suppress TARC levels by 30% to 70%. In contrast, dupilumab did not significantly inhibit TARC levels in the same assay.

[0237] Figure 13 shows that FB759 inhibits HDM-induced TARC levels in PBMCs, and that dupilumab does not or minimally inhibits TARC in the same assay. For the data shown in Figure 13, hIgG4 (human IgG4 isotype Ab), BFB759, and dupilumab were used at 30 μg / ml, and HDM was used at 10 μg / ml. Data shown are mean ± SD.

[0238] Figure 14 shows a summary of the inhibition of HDM-induced TARC levels in PBMCs from different donors by BFB759. For the data shown in Figure 14, HDM was used at 10 μg / ml, BFB759 at 1-30 μg / ml, and PBMCs were obtained from 12 different donors. Data shown are mean ± SD.

[0239] Example 9. TARC inhibitory effects of BFB759 and dupilumab in PBMCs treated with HDM and hIL-4

[0240] Experiments were conducted using BFB759 and dupilumab to characterize the inhibition of TARC production in PBMCs treated with HDM and hIL-4.

[0241] Methods and Results Dupilumab is approved for the treatment of AD (atopic dermatitis). Dupilumab inhibits IL-4 / 13 signaling by blocking the binding of IL-4 / 13 to IL-4Ra. Stimulation of PBMCs with HDM and IL-4 significantly increased TARC expression. Treatment of cells with BFB759 or dupilumab alone partially inhibited TARC expression in the HDM and IL-4 model. These results also demonstrate that the combined effect of BFB759 and dupilumab was greater than the effect of each agent alone, regardless of the concentration tested. Near-complete inhibition of TARC expression was achieved when these antibodies were combined at 10 μg / ml per antibody. These results were unexpected, as reduction of TARC through TH1 cytokine inhibition with therapeutic antibodies such as BFB759 has not been reported. Furthermore, it was surprising to find that while BFB759 at 1 or 0.1 μg / ml only minimally inhibited TARC expression, when combined with dupilumab, this combination was even more effective than 10 μg / ml of dupilumab alone (i.e., up to a 100-fold reduction in dupilumab), suggesting that combination therapy with BFB759 may allow for a significant reduction in dupilumab dose in clinical settings, while simultaneously improving efficacy and widening the therapeutic window (Figure 15).

[0242] Figure 15 shows that BFB759 and dupilumab can inhibit TARC levels in PBMCs treated with HDM and IL-4. For the data shown in Figure 15, HDM was used at 10 μg / ml, IL-4 at 750 pg / ml, and antibodies were used at three different concentrations as indicated on the graph. Data shown are means ± SD.

[0243] Example 10.

[0244] Inhibitory effect of BFB759 and anti-IL-13 on TARC levels in PBMCs treated with HDM and IL-13.

[0245] Experiments were performed using BFB759 and anti-IL-13 to characterize the inhibition of TARC production in PBMCs treated with HDM and IL-13. The following method was used.

[0246] Methods and Results The IL-13 neutralizing antibody tralokinumab was approved for the treatment of atopic dermatitis (AD) in 2021. Stimulation of PBMCs with HDM and IL-13 significantly increased TARC levels.

[0247] Treatment of HDM and IL-13-stimulated cells with tralokinumab inhibited TARC in a dose-dependent manner. Combining tralokinumab with 1 μg / ml BFB759 further reduced TARC levels, indicating that the combined effect of BFB759 and tralokinumab was greater than the effect of either drug alone, regardless of the concentration tested. These results were unexpected, as there have been no reports of TARC reduction through TH1 cytokine inhibition with therapeutic antibodies such as BFB759 (Figure 16). Furthermore, these results are surprising, as there have been no reports of TARC expression reduction through combination therapy with anti-IL-13 and anti-IL-1R3 antibodies.

[0248] Figure 16 shows that BFB759 and tralokinumab can inhibit TARC levels in PBMCs treated with HDM and IL-13. For the data shown in Figure 16, HDM was used at 10 μg / ml, IL-13 at 0.5 nM, BFB759 at 1 μg / ml, and tralokinumab was gradually titrated from 10 μg / ml to 0.16 μg / ml (four-fold dilutions, four series). Data shown are mean ± SD.

[0249] Example 11

[0250] Effect of TARC inhibition by BFB759 and JAK inhibitors or dexamethasone in HDM-treated PBMCs.

[0251] Experiments were conducted to characterize the inhibition of TARC production in HDM-treated PBMCs using BFB759 and a JAK inhibitor, or dexamethasone.

[0252] Methods and Results JAK inhibitors (abrocitinib and upadacitinib) and dexamethasone are used to treat AD patients. Treatment of HDM-stimulated cells with a JAKi or dexamethasone dose-dependently inhibited TARC. Combining a JAKi or dexamethasone with 1 μg / ml BFB759 further reduced TARC levels, indicating that the combined effect of BFB759 and these agents was greater than the effect of either agent alone, regardless of the concentration tested. These results were unexpected, as TARC reduction by TH1 cytokine inhibition with therapeutic antibodies such as BFB759 has not been reported (Figures 17A and 17B). Furthermore, these results are surprising, as reduction of TARC expression by combination therapy with a JAKi and dexamethasone with an anti-IL-1R3 antibody has not been reported (Figures 17A, 17B, 18A, and 18B).

[0253] These results suggest that combination therapy with BFB759 could significantly reduce the dose of JAKi or dexamethasone in clinical settings, while improving efficacy and providing a broader therapeutic window.

[0254] Figures 17A and 17B show that the combination of BFB759 with a JAK inhibitor or dexamethasone can inhibit TARC in HDM-treated PBMCs. For the data shown in Figures 17A and 17B, HDM was used at 10 μg / ml, BFB759 was used at 1 μg / ml, and the JAKi and dexamethasone were tested at three different concentrations (0.1 nM, 10 nM, and 1 μM) as indicated on the graphs. Data shown are mean ± SD.

[0255] Figures 18A and 18B show that the combination of BFB759 and upadacitinib inhibits TARC in PBMCs treated with HDM. For the data shown in Figures 18A and 18B, M was used at 10 μg / ml, BFB759 and hIgG4 (human IgG4 isotype Ab) were used at 1 μg / ml, and the JAKi upadacitinib was tested in a 5-fold decrement series from 1 μM to 0.01 nM. Data shown are mean ± SD.

[0256] Example 12 Combined inhibitory effect of BFB759 and adalimumab on IL-6 production in human whole blood cultures stimulated with heat-killed Candida albicans

[0257] Experiments were conducted to characterize the inhibition of IL-6 production by the combination of BFB759 and adalimumab in human whole blood cultures stimulated with heat-killed Candida albicans.

[0258] method The materials included human peripheral blood collected from five healthy donors, anti-human TNF-alpha (Adalimumab Biosimilar), BioXCell (Cat. #SIM0001, Lot #8024221F1), heat-killed Candida albicans (HKCA) (InvivoGen, Cat. #tlrl-hkca, Lot #6292-44-02), RPMI 1640 with L-glutamine and HEPES (Corning, Cat. #10-041-CM), and Human IL-6 DuoSet ELISA (R and D Systems, Cat. #DY206-05).

[0259] Blood was collected from healthy volunteers in heparinized tubes. Whole blood was diluted with RPMI 1640 medium and placed in sterile 5.0 ml polypropylene snap-cap culture tubes. One of the following treatments was added 1 hour before stimulation with 1.5 x 10^6 heat-killed Candida albicans cells: medium alone (cells alone), 1 μg / ml IgG1 isotype control antibody alone, 1 μg / ml BFB759 alone, 1 μg / ml adalimumab alone, or one of two combination treatments: 1 μg / ml IgG1 isotype control antibody + 0.5 μg / ml adalimumab, or 1 μg / ml BFB759 + 0.5 μg / ml adalimumab. The final volume of each culture was 1.0 ml. Incubation was continued for 24 hours at 37°C / 5% CO2. Replicate supernatants from each culture were stored frozen at −80°C until repeated measurements of IL-6 levels were performed by ELISA.

[0260] result BFB759 and adalimumab inhibited IL-6 secretion in human whole blood cultures challenged with HKCA, and this effect was enhanced by the combination of the two treatments. High levels of IL-6 were induced in blood cultures after challenge with HKCA. Both adalimumab and BFB759 could suppress IL-6 secretion, and the effect was enhanced by the combination of the two treatments.

[0261] Figures 19A and 19B show that the combination of BFB759 and adalimumab inhibits IL-6 secretion in human whole blood cultures stimulated with HKCA. For the data shown in Figure 19A, IL-6 levels from whole blood cultures are measured by ELISA. For the data shown in Figure 19B, the percentage of IL-6 measured in cultures treated as indicated compared to the levels detected with HKCA without intermediate treatment.

[0262] Example 13 Inhibitory effect of BFB759 and secukinumab on IL-6 and IL-8 production in normal human dermal fibroblasts (NHDFs) stimulated with a combination of cytokines

[0263] Experiments were conducted to characterize the inhibition of IL-6 and IL-8 production in normal human dermal fibroblasts stimulated with a combination of cytokines by the combination of BFB759 and secukinumab. [Table 7]

[0264] method Human NHDF cells were plated at 4,000 cells / well in 96-well flat-bottom plates. BFB759, secukinumab (anti-IL-17A antibody), and increasing concentrations of an isotype control antibody (0-20 μg / ml, 5-fold dilutions) were incubated with the cells in duplicate for 45 minutes. A combination of cytokines [IL-1α (10 pg / ml), IL-1β (5 pg / ml), IL-36α (25 ng / ml), IL-36β (3 ng / ml), IL-36γ (7 ng / ml), and IL-17A (1 ng / ml)] was then added to the cells' EC. 50 The cells were added at concentrations previously determined to be approximately equivalent (data not shown). For combination therapy, 0.5 μg / ml of BFB759 was mixed with increasing concentrations (0–20 μg / ml, 5-fold dilutions) of secukinumab or an isotype control antibody and incubated with the cells in duplicate for 45 minutes before the addition of the cytokine combination. The plates were then incubated overnight at 37°C and 5% CO2. After 24 hours of treatment, supernatants were transferred and assayed for IL-6 and IL-8 according to the manufacturer's protocol (R&D Systems; Human IL-6 DuoSet ELISA, Human IL-8 / CXCL8 DuoSet ELISA). Raw assay data were analyzed using GraphPad Prism 9 software, performing nonlinear regression.

[0265] result NHDF cells are known to respond to IL-1, IL-36, and IL-17A stimulation. To understand whether the combination of BFB759 and secukinumab could further inhibit IL-6 and IL-8 production, NHDF cells were first cultured with BFB759, secukinumab, and the combination of BFB759 and secukinumab. They were then stimulated with a cytokine combination consisting of IL-1α / β, IL-36α / β / γ, and IL-17A. Treatment of NHDFs with BFB759 dose-dependently reduced IL-6 and IL-8 cytokine production (Figure 20). Secukinumab alone only limitedly inhibited IL-6 and IL-8 release in this assay. Interestingly, the combination of BFB759 (0.5ug / ml) with high concentrations of secukinumab suppressed IL-6 and IL-8 production more than either antibody alone, demonstrating the utility of combination therapy in autoimmune and inflammatory diseases.

[0266] Figures 20A and 20B show that the combination of BFB759 and secukinumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with the cytokine combination. Data shown are mean ± SD.

[0267] Example 14. Inhibitory effect of BFB759 and adalimumab in combination on IL-6 and IL-8 production in normal human dermal fibroblasts (NHDFs) stimulated with a combination of cytokines.

[0268] Experiments were conducted to characterize the suppression of IL-6 and IL-8 production by the combination of BFB759 and adalimumab in normal human dermal fibroblasts stimulated with a combination of cytokines. [Table 8]

[0269] method Human NHDF cells were plated at 4,000 cells / well in 96-well flat-bottom plates. BFB759, adalimumab (anti-TNF-α antibody), and increasing concentrations of an isotype control antibody (0-20 μg / ml, 5-fold dilutions) were incubated with the cells in duplicate for 45 minutes. A combination of cytokines [IL-1α (10 pg / ml), IL-1β (5 pg / ml), IL-36α (25 ng / ml), IL-36β (3 ng / ml), IL-36γ (7 ng / ml), and TNF-α (0.05 ng / ml)] was then added to the EC of the cells. 50 BFB759 was added at concentrations previously determined to be approximately equivalent (data not shown). For combination therapy, 0.5 μg / ml of BFB759 was mixed with increasing concentrations of adalimumab or an isotype control antibody (0–20 μg / ml, 5-fold dilutions) and incubated with cells in duplicate for 45 minutes before the addition of the cytokine combination. Plates were then incubated overnight at 37°C and 5% CO2. After 24 hours of treatment, supernatants were transferred and assayed for IL-6 and IL-8 according to the manufacturer's protocol (R&D Systems; Human IL-6 DuoSet ELISA, Human IL-8 / CXCL8 DuoSet ELISA). Raw assay data were analyzed using GraphPad Prism 9 software, performing nonlinear regression.

[0270] result NHDF cells are known to respond to IL-1, IL-36, and TNF-α stimulation. To understand whether the combination of BFB759 and adalimumab could further inhibit IL-6 and IL-8 production, NHDF cells were first cultured with BFB759, adalimumab, and the combination of BFB759 and adalimumab. They were then stimulated with a combination of cytokines consisting of IL-1α / β, IL-36α / β / γ, and IL-17A. Treatment of NHDFs with BFB759 dose-dependently reduced IL-6 and IL-8 cytokine production (Figure 21). Adalimumab alone only limitedly inhibited IL-6 and IL-8 release in this assay. Interestingly, the combination of BFB759 (0.5µg / ml) with high concentrations of adalimumab suppressed IL-6 and IL-8 production more than either antibody alone, demonstrating the utility of combination therapy in autoimmune and inflammatory diseases.

[0271] Figures 21A and 21B show that the combination of BFB759 and adalimumab inhibits the release of IL-6 and IL-8 in NHDF cells stimulated with the cytokine combination. Data shown are mean values ± SD.

[0272] Incorporation by Reference The contents of all references, patents, pending patent applications, published patents, sequence listings, and accession numbers cited throughout this application are hereby expressly incorporated by reference.

[0273] equivalent 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 which equivalents are intended to be encompassed by the following claims. [Table 9-1] [Table 9-2]

Table 9-3

Table 9-4

Table 9-5

Table 9-6

Table 9-7

Table 9-8

Table 9-9

Claims

1. A method of treating an inflammatory or autoimmune condition in a mammal in need thereof, comprising administering a therapeutically effective amount of an antibody against IL1RAP.

2. 2. The method of claim 1, wherein the inflammatory or autoimmune condition is an atopic condition and the antibody reduces the level of TARC / CCL17, PARC, periostin, IL-22, eotaxin-1, eotaxin-3, or a combination thereof at an atopic site or in serum.

3. 2. The method of claim 1, wherein the inflammatory or autoimmune condition is characterized by or caused by an increased number of neutrophil or eosinophil cells, neutrophil or eosinophil dysfunction, or increased TARC / CCL17 levels.

4. 4. The method of claim 3, wherein the inflammatory or autoimmune condition is characterized or caused by elevated neutrophil cell numbers or neutrophil dysfunction.

5. 5. The method of claim 4, wherein the inflammatory or autoimmune condition characterized by or caused by elevated neutrophil cell count or neutrophil dysfunction is selected from the group consisting of hidradenitis suppurativa, generalized pustular psoriasis (GPP), COPD, idiopathic fibrosis, neutrophilic asthma, neutrophilic dermatosis, pyoderma gangrenosum-Schnitzler syndrome, Behcet's disease, Sweet's syndrome, rheumatoid arthritis, systemic lupus erythematosus (SLE), inflammatory bowel disease (Crohn's disease, ulcerative colitis), psoriasis, vasculitis, Alzheimer's disease, COVID-19, and gout.

6. 4. The method of claim 3, wherein the inflammatory or autoimmune condition is further characterized by an increased number of eosinophil cells or eosinophil dysfunction.

7. 7. The method of claim 6, wherein the inflammatory or autoimmune condition characterized by or caused by increased eosinophil cell count or eosinophil dysfunction is selected from the group consisting of atopic dermatitis (eczema), allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, and eosinophilic myocarditis.

8. 4. The method of claim 3, wherein the inflammatory or autoimmune condition is further characterized by increased levels of TARC / CCL17.

9. The method of claim 8, wherein the inflammatory or autoimmune condition characterized by increased TARC / CCL17 levels is selected from the group consisting of atopic dermatitis, COPD, bronchial asthma, allergic rhinitis, eosinophilic pneumonia, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, mastocytosis, and eosinophil-associated diseases.

10. The inflammatory or autoimmune condition is selected from the group consisting of sepsis, acute respiratory distress syndrome, COVID-19, myocardial infarction, cystic fibrosis, irritable bowel disease, ulcerative colitis, Crohn's disease, atopic dermatitis, psoriasis, multiple sclerosis, asthma, neutrophilic asthma, Alzheimer's disease, stroke, diabetic kidney disease, diabetic retinopathy, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, non-alcoholic fatty liver disease, rheumatoid arthritis, psoriatic arthritis, ankylosing leukemia, and rheumatoid arthritis.

2. The method of claim 1, wherein the disease is selected from the group consisting of spondylitis, axonal spondyloarthritis, systemic lupus erythematosus (SLE), vasculitis, gout, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorders, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, and mastocytosis.

11. 10. The method of claim 1, further comprising identifying a mammal in need thereof according to an abnormal serum level of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof.

12. 10. The method of claim 1, further comprising identifying a mammal in need thereof according to abnormal serum levels of CXCL1, CXCL8, LCN-2, TARC, or a combination thereof.

13. 12. The method of claim 11, wherein the serum level of IL-1α, IL-1β, IL-33, IL-36α, IL-36β, IL-36γ, or a combination thereof is elevated.

14. 13. The method of claim 12, wherein the serum levels of CXCL1, CXCL8, LCN-2, TARC, or a combination thereof are elevated.

15. 2. The method of claim 1, wherein the antibody is characterized by inhibiting IL-8 release by intestinal epithelial cells, intestinal myofibroblasts, or skin fibroblasts stimulated with IL-1α, IL-1β, IL-36α, IL-36β, and / or IL-36γ, or a combination thereof.

16. 3. The method of claim 2, wherein the atopic condition is selected from the group consisting of atopic dermatitis, asthma, COPD, allergic rhinitis, allergic conjunctivitis, food allergy, drug allergy, and angioedema.

17. 3. The method of claim 2, wherein the atopic condition is atopic dermatitis and the antibody reduces the level of TARC / CCL17, PARC, periostin, IL-22, eotaxin-1, eotaxin-3, or a combination thereof in the skin or serum.

18. 3. The method of claim 2, wherein the atopic condition is asthma.

19. 3. The method of claim 2, wherein the atopic condition is COPD.

20. The method of claim 1 , wherein the antibody is administered in combination with at least one other therapeutic agent.

21. 21. The method of claim 20, wherein the at least one other therapeutic agent is a therapeutic antibody, a corticosteroid, a small molecule, an siRNA, an mRNA, or a combination thereof.

22. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-4 signaling.

23. 23. The method of claim 22, wherein the at least one other therapeutic agent is an IL-4Rα inhibitor or antagonist, a pan-JAK inhibitor or antagonist, or a combination thereof.

24. 23. The method of claim 22, wherein the at least one other therapeutic agent is dupilumab, CBP-201, AK120, celdulatinib, CEE321, yaktinib, delgocitinib, filgotinib, tofacitinib, dexamethasone, triamcinolone, prednisone, or a combination thereof.

25. 23. The method of claim 22, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, polyarticular juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

26. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-13 signaling.

27. 27. The method of claim 26, wherein the at least one other therapeutic agent is an IL-13 inhibitor or antagonist, a JAK1 inhibitor or antagonist, a TYK2 inhibitor or antagonist, or a combination thereof.

28. 27. The method of claim 26, wherein the at least one other therapeutic agent is upadacitinib, abrocitinib, tralokinumab, lebrikizumab, ebrasakimab, baricitinib, ruxolitinib, filgotinib, PF-06651600, dexamethasone, triamcinolone, prednisone, or a combination thereof.

29. 27. The method of claim 26, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, systemic lupus erythematosus, vitiligo, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

30. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-22 signaling.

31. 31. The method of claim 30, wherein the at least one other therapeutic agent is an IL-22 inhibitor or antagonist, an IL-22R1 inhibitor or antagonist, a JAK1 inhibitor or antagonist, a JAK2 inhibitor or antagonist, a TYK2 inhibitor or antagonist, or a combination thereof.

32. 31. The method of claim 30, wherein the at least one other therapeutic agent is fezakinumab, LEO138559, brepositinib, ATI-1777, deuclavacitinib, TAK-279, upadacitonib, and abrocitinib, or a combination thereof.

33. 31. The method of claim 30, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, systemic lupus erythematosus, vitiligo, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

34. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-33 signaling.

35. 35. The method of claim 34, wherein the at least one other therapeutic agent is an IL-33 or IL-33R inhibitor or antagonist.

36. 35. The method of claim 34, wherein the at least one other therapeutic agent is etokimab, itepekimab, astegolimab, PF-06817024, tozorakimab, CNTO 7160, or a combination thereof.

37. 35. The method of claim 34, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, chronic obstructive pulmonary disease (COPD), asthma, allergic contact dermatitis, irritant contact dermatitis, rosacea, plaque psoriasis, pustular psoriasis, mastocytosis, systemic lupus erythematosus, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, Behcet's disease, or vitiligo.

38. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-17 signaling.

39. 39. The method of claim 38, wherein the at least one other therapeutic agent is an IL-17A, IL-17C, IL-17F, IL17A / F, or IL-17RA inhibitor or antagonist, or a combination thereof.

40. 39. The method of claim 38, wherein the at least one other therapeutic agent is secukinumab, ixekizumab, brodalumab, bimekizumab, izokibep, sonelokimab, or a combination thereof.

41. 39. The method of claim 38, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, tendonitis-associated arthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COPD, or asthma.

42. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-36 signaling.

43. 43. The method of claim 42, wherein the at least one other therapeutic agent is an IL-36 inhibitor or antagonist, or an IL-36R inhibitor or antagonist.

44. 43. The method of claim 42, wherein the at least one other therapeutic agent is spesolimab, imsidolimab, REGN6490, or a combination thereof.

45. The inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, neutrophilic asthma, neutrophilic pneumonia, COVID-19, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, nonalcoholic fatty liver disease, neutrophilic dermatosis, hidradenitis suppurativa, generalized pustular psoriasis, and rheumatoid arthritis.

43. The method of claim 42, wherein the skin irritation is caused by psoriasis, palmoplantar pustular psoriasis, IL-36 receptor antagonist (DITRA) deficiency, plaque psoriasis, CARD14-mediated psoriasis, acute generalized exanthematous pustulosis, pyoderma gangrenosum, Sweet's syndrome, systemic lupus erythematosus, systemic sclerosis, autoimmune bullous disease, acne, allergic contact dermatitis, or folliculitis and eosinophilic pustular folliculitis.

46. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-18 signaling.

47. 47. The method of claim 46, wherein the at least one other therapeutic agent is an IL-18 inhibitor or antagonist.

48. 47. The method of claim 46, wherein the at least one other therapeutic agent is tadequinig alfa.

49. 47. The method of claim 46, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, asthma, adult-onset Still's disease, cutaneous lupus erythematosus, chronic spontaneous urticaria, contact dermatitis, alopecia areata, cutaneous drug eruption, graft-versus-host disease, cryopyrin-associated periodic syndrome, granulomatosis with polyangiitis, systemic sclerosis, hidradenitis suppurativa, septic arthritis, pyoderma gangrenosum and acne (PAPA), familial Mediterranean fever, rosacea, synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO), bullous pemphigoid, pemphigus vulgaris, Behcet's disease, or Schnitzler's syndrome.

50. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-23 signaling.

51. 51. The method of claim 50, wherein the at least one other therapeutic agent is an IL-23 inhibitor or antagonist, an IL-12 / 23p40 subunit inhibitor or antagonist, an IL-23p19 subunit inhibitor or antagonist, or a combination thereof.

52. 51. The method of claim 50, wherein the at least one other therapeutic agent is risankizumab, ustekinumab, guselkumab, tildrakizumab, mirikizumab, brazikumab, or a combination thereof.

53. 51. The method of claim 50, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

54. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits OX40 signaling.

55. 55. The method of claim 54, wherein the at least one other therapeutic agent is an OX40 or OX40L inhibitor or antagonist.

56. 55. The method of claim 54, wherein the at least one other therapeutic agent is locatinlimab, GBR830, amritelimab, or a combination thereof.

57. 55. The method of claim 54, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

58. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IL-5 signaling.

59. 59. The method of claim 58, wherein the at least one other therapeutic agent is an IL-5Rα inhibitor or antagonist, a JAK1 inhibitor or antagonist, or a combination thereof.

60. 59. The method of claim 58, wherein the at least one other therapeutic agent is benralizumab, upadacitinib, abrocitinib, SHR0302, filgotinib, PF-06651600, dexamethasone, triamcinolone, prednisone, or a combination thereof.

61. 59. The method of claim 58, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

62. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits T cell migration.

63. 63. The method of claim 62, wherein the at least one other therapeutic agent is an S1PR1 inhibitor or antagonist, an S1PR4 inhibitor or antagonist, an S1PR5 inhibitor or antagonist, a CCR4 inhibitor or antagonist, or a combination thereof.

64. 63. The method of claim 62, wherein the at least one other therapeutic agent is etrasimod, ozanimod, SCD-044, LC51-0255, BMS-986166, RPT193, or a combination thereof.

65. 63. The method of claim 62, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

66. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits pruritus.

67. 67. The method of claim 66, wherein the at least one other therapeutic agent is an IL-1α inhibitor or antagonist, an OSMRβ inhibitor or antagonist, an NK1R inhibitor or antagonist, a P2X3 inhibitor or antagonist, an IL-31 inhibitor or antagonist, or a combination thereof.

68. 67. The method of claim 66, wherein the at least one other therapeutic agent is bermekimab, bicsarelimab, serlopitant, tradipitant, BLU-5937, nemolizumab, or a combination thereof.

69. 67. The method of claim 66, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, or asthma.

70. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits a TH1-associated immune response.

71. 71. The method of claim 70, wherein the at least one other therapeutic agent is an IL-1α inhibitor or antagonist, an IL-1β inhibitor or antagonist, an IL-1R1 inhibitor or antagonist, an IL-36R inhibitor or antagonist, a TNFα inhibitor or antagonist, or a combination thereof.

72. 71. The method of claim 70, wherein the at least one other therapeutic agent is bermekimab, anakinra, canakinumab, gevokizumab, rilonacept, MEDI8968, spesolimab, imsidolimab, REGN6490, adalimumab, infliximab, etanercept, certolizumab, golimumab, or a combination thereof.

73. The inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, familial Mediterranean fever (FMF), and septic arthritis. , pyoderma gangrenosum, acne (PAPA), cryopyrin-associated periodic syndromes (CAPS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndromes (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis ulcerative colitis, panniculitis, Erdheim-Chester syndrome, adenosine deaminase deficiency (DADA2), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune 71. The method of claim 70, wherein the skin disease is epidermal bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratosis (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), neutrophilic dermatosis, or a solitary autoinflammatory skin disease.

74. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits a TH2-associated immune response.

75. 75. The method of claim 74, wherein the at least one other therapeutic agent is an IL-4 inhibitor or antagonist, a type I IL-4 receptor inhibitor or antagonist, a type II IL-4 receptor inhibitor or antagonist, an IL-13 inhibitor or antagonist, a type I IL-13 receptor inhibitor or antagonist, a type II IL-13 receptor inhibitor or antagonist, and an IL-5 inhibitor or antagonist, a type I IL-5 receptor inhibitor or antagonist, a type II IL-5 receptor inhibitor or antagonist, an IL-9 inhibitor or antagonist, a type I IL-9 receptor inhibitor or antagonist, an IL-10 inhibitor or antagonist, a homodimeric IL-10 receptor inhibitor or antagonist, a heterodimeric IL-10 receptor inhibitor or antagonist, or a combination thereof.

76. 75. The method of claim 74, wherein the at least one other therapeutic agent is dupilumab, omalizumab, mepolizumab, benralizumab, tralokinumab, lebrikizumab, or a combination thereof.

77. The inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, familial Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, acne (PAPA), and cryopyrin-associated periodontitis. Hyper IgD syndrome (HIDS), also known as Capsular Adenosine Phosphate Syndrome (CAPS), Mevalonate Kinase Deficiency (MKD), TNF Receptor-Associated Periodic Syndrome (TRAPS), IL-1 Receptor Antagonist Deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, Generalized Pustular Psoriasis (GPP), Palmoplantar Pustular Psoriasis (PPP), Dermatomyositis, Panniculitis, Erdheim-Chester syndrome, Adenosine Deaminase Deficiency (DADA2), Majeed syndrome, IL-3 6 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratoses (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammatory disease with neutrophilic dermatosis 75. The method of claim 74, wherein the inflammatory disease is selected from the group consisting of eosinophilic granulomatosis (PAAND), NLRC4-associated macrophage activation syndrome (NLRC4-MAS), neutrophilic dermatosis or isolated autoinflammatory skin disease, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, and urticaria.

78. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits a TH17-associated immune response.

79. 79. The method of claim 78, wherein the at least one other therapeutic agent is an IL-17 inhibitor or antagonist, a type I IL-17 receptor inhibitor or antagonist, a type II IL-17 receptor inhibitor or antagonist, an IL-23 inhibitor or antagonist, an IL-23 receptor inhibitor or antagonist, or a combination thereof.

80. 79. The method of claim 78, wherein the at least one other therapeutic agent is secukinumab, ixekizumab, brodalumab, bimekizumab, izokibep, sonelokimab, risankizumab, ustekinumab, guselkumab, tildrakizumab, mirikizumab, brazikumab, or a combination thereof.

81. The inflammatory or autoimmune condition is selected from the group consisting of rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, COPD, asthma, adult-onset Still's disease, Behcet's disease, hidradenitis suppurativa, pyoderma gangrenosum, SAPHO, acne vulgaris, psoriasis vulgaris, Schnitzler's syndrome, urticarial vasculitis, familial Mediterranean fever (FMF), septic arthritis, pyoderma gangrenosum, acne (PAPA), cryopyrin-associated periodic syndrome (CPS), and / or rheumatoid arthritis (RH). APS), hyper-IgD syndrome (HIDS), also known as mevalonate kinase deficiency (MKD), TNF receptor-associated periodic syndrome (TRAPS), IL-1 receptor antagonist deficiency (DIRA), Sweet's syndrome, PASH, PFAPA, generalized pustular psoriasis (GPP), palmoplantar pustular psoriasis (PPP), dermatomyositis, panniculitis, Erdheim-Chester syndrome, adenosine deaminase 2 (DADA2), Majeed syndrome, IL-36 receptor antagonist deficiency (DITRA), haploinsufficiency of A20 (HA20), PAPASH, rosacea, acute generalized exanthematous pustulosis (AGEP), allergic contact dermatitis, irritant contact dermatitis, mastocytosis, systemic sclerosis, chronic spontaneous urticaria, autoimmune bullous disease, vitiligo, CARD-14-mediated pustular psoriasis (CAMPS), familial chronic lichenoid keratoses (FKLC), multiple self-healing palmoplantar carcinoma (MSPC), pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND), NLR 79. The method of claim 78, wherein the condition is C4-associated macrophage activation syndrome (NLRC4-MAS), neutrophilic dermatosis, or isolated autoinflammatory skin disease, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis (EoE), eosinophilic asthma, hypereosinophilic syndrome (HES), eosinophilic granulomatosis, eosinophilic fasciitis, eosinophilic gastrointestinal disorder, eosinophilic pneumonia, eosinophilic myocarditis, hypersensitivity pneumonitis, lichen planus, sarcoidosis, urticaria, or isolated autoinflammatory skin disorder.

82. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits IRAK4 signaling.

83. 83. The method of claim 82, wherein the at least one other therapeutic agent is an IRAK4 inhibitor, degrader, or antagonist.

84. 83. The method of claim 82, wherein the at least one other therapeutic agent is PF-06650833, CA-4948, KT-474, or a combination thereof.

85. 83. The method of claim 82, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COPD, or asthma.

86. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits complement signaling.

87. 87. The method of claim 86, wherein the at least one other therapeutic agent is a C5a inhibitor or antagonist, a C5aR inhibitor or antagonist, a TSLP inhibitor or antagonist, a CD80 / CD86 inhibitor or antagonist, an IL-6 inhibitor or antagonist, a CD20 inhibitor or antagonist, an integrin alpha 4 inhibitor or antagonist, an AhR agonist, or a combination thereof.

88. 87. The method of claim 86, wherein the at least one other therapeutic agent is vilobelimab, FX002, INF904, tezepelumab, abatacept, tocilizumab, sarilumab, rituximab, vedolizumab, tapinarof, tadekinig alfa, or a combination thereof.

89. 87. The method of claim 86, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, hidradenitis suppurativa, COVID-19, COPD, or asthma.

90. 21. The method of claim 20, wherein the at least one other therapeutic agent inhibits PDE4 signaling.

91. 91. The method of claim 90, wherein the at least one other therapeutic agent is a PDE4 inhibitor or antagonist.

92. 91. The method of claim 90, wherein the at least one other therapeutic agent is apremilast, crisaborole, difamilast, roflumilast, or a combination thereof.

93. 91. The method of claim 90, wherein the inflammatory or autoimmune condition is rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, axial spondyloarthritis, Crohn's disease, ulcerative colitis, atopic dermatitis, pustular purpura, COPD, or asthma.

94. 21. The method of claim 20, wherein the at least one other therapeutic agent is administered before, simultaneously with, or after the at least one other therapeutic agent.

95. 10. The method of claim 1, wherein the antibody is administered subcutaneously or intravenously at a dose selected from 1-80 mg / kg, 1-60 mg / kg, 1-50 mg / kg, 1-40 mg / kg, 1-30 mg / kg, 1-25 mg / kg, or 1-20 mg / kg.

96. 18. The method of claim 17, wherein the antibody is administered subcutaneously or intravenously at a dose selected from 1-80 mg / kg, 1-60 mg / kg, 1-50 mg / kg, 1-40 mg / kg, 1-30 mg / kg, 1-25 mg / kg, or 1-20 mg / kg.

97. 21. The method of claim 20, wherein the antibody is administered subcutaneously or intravenously at a dose selected from 1-80 mg / kg, 1-60 mg / kg, 1-50 mg / kg, 1-40 mg / kg, 1-30 mg / kg, 1-25 mg / kg, or 1-20 mg / kg.

98. 10. The method of claim 1, wherein the antibody is administered as a subcutaneous injection or an intravenous bolus less frequently than once a week, twice a week, more frequently than twice a week, or by continuous infusion.

99. 18. The method of claim 17, wherein the antibody is administered as a subcutaneous injection or an intravenous bolus less frequently than once a week, twice a week, more frequently than twice a week, or by continuous infusion.

100. 21. The method of claim 20, wherein the antibody is administered as a subcutaneous injection or an intravenous bolus less frequently than once a week, twice a week, more frequently than twice a week, or by continuous infusion.

101. The antibody a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 67, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 66, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 70, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 69; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 15, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 51, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 50, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 49, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 55, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 54, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 53; a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 59, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 58, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 57, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 63, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 62, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 61; or BFB759 (i.e., 37E10_15B5) corresponding to a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 176, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 175, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 174, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 179, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 178, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 79, 21. The method of claim 1, 17, or 20.

102. The antibody a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 68; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 82; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 48 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 52; a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 56 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 60; or a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 173 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 177; 21. The method of claim 1, 17, or 20.

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