Anti-IL1RAP Antibodies

Abstract

Anti-IL1RAP antibodies for diagnostic and therapeutic uses are provided. The present invention relates to anti-IL1RAP binding compounds, particularly novel anti-IL1RAP antibodies, and therapeutic and diagnostic methods and compositions for their use.

Description

[Technical field]

[0001] Sequence Listing This application has been submitted electronically in ASCII format and contains a Sequence Listing, which is hereby incorporated by reference in its entirety. Said ASCII copy, created on February 3, 2022, is entitled 09-0717-WO-1_SL.txt and is 194,305 bytes in size.

[0002] The present invention relates generally to anti-IL1RAP antibodies for diagnostic and therapeutic use. The antibodies may be used in pharmaceutical compositions and kits containing such compounds. The antibodies are useful in methods for the treatment of various diseases or disorders, such as immunological, inflammatory, autoimmune, fibrotic, and respiratory diseases in humans. [Background technology]

[0003] The IL-1 family of cytokines consists of 11 different ligands, namely IL-1α (also called IL-1F1), IL-1β (IL-1F2), IL-1 receptor antagonist (IL-1Ra or IL-1F3), IL-18 (IL-1F4), IL-1F5-IL-1F10, and IL-1F11 (or IL-33). IL-1α and IL-1β are known to induce proinflammatory activity upon binding to the type I IL-1 receptor (IL-1RI) and recruitment of the common co-receptor IL-1 receptor accessory protein (IL-1RAcP), whereas IL-1Ra acts as a competitive inhibitor of IL-1 binding to IL-1RI and thus exerts anti-inflammatory activity. Numerous studies have reported that IL-18 is a proinflammatory cytokine that is an inducer of IFN-γ, whereas IL-33 has been described as an immunoregulatory cytokine specifically involved in the control of Th2 responses.

[0004] The IL1 receptor accessory protein IL1RAP is a coreceptor for the major receptors (IL1R1, IL33R, and IL36R) required for signaling of the cognate IL1 family cytokines IL1 (α, β), IL33, and IL36 (α, β, γ). The IL1 family cytokines IL1α and IL1β, IL33, and IL36α, IL36β, and IL36γ bind to their respective receptors IL1R1, IL33R, and IL36R. The cognate receptors then bind to the IL1 receptor accessory protein IL1RAP (also designated IL-1RAP), which acts as a coreceptor leading to downstream activation mediated by IL1, IL33, and IL36, respectively. IL1RAP is expressed in tissues where IL1-, IL33-, or IL36-receptors are present, such as lymph nodes (thymus, tonsils), bone marrow, brain, lung, skin, gastrointestinal tract, liver, and placenta. Myeloid leukemia stem cells in CML reportedly also express IL1RAP (Jaras, et al. (2010). Isolation and killing of candidate chronic myeloid leukemia stem cells by antibody targeting of IL-1 receptor accessory protein. Proc Natl Acad Sci USA 107, 16280-16285). The importance of IL1RAP in signaling of the IL1 family of alarmins has been demonstrated by genetic deletion of the co-receptor IL1RAP, which led to a complete loss of signaling for IL1 (IL-1 ip injection (Cullinan, et al. (1998). IL-1 receptor accessory protein is an essential component of the IL-1 receptor. J Immunol 161, 5614-5620)); and IL33 (mast cells (Palmer, et al. (2008). The IL-1 receptor accessory protein (AcP) is required for IL-33 signaling and soluble AcP enhances the ability of soluble ST2 to inhibit IL-33. Cytokine 42, 358-364.)), as well as by transfection studies / pharmacological intervention for IL36 (Towne, et al. (2011). Interleukin-36 (IL-36) ligands require processing for full agonist (IL-36alpha, The inhibitors of IL-36β, IL-36βgamma, and IL-36Ra have been confirmed by IL-36β antagonist (IL-36β, IL-36ββ, and IL-36βgamma) or antagonist (IL-36Ra) activity. J Biol Chem 286, 42594-42602). Individual cytokine inhibitors have been well reported, however, when tested in vivo in clinical trials, these inhibitors have failed, implying that a single neutralizing antibody against one or two IL1 cytokines may not be sufficient for the desired clinical efficacy.

[0005] Only a few IL1RAP antibodies have been described in the published literature. Of note, most of these antibodies only achieve a certain level of biological activity against some of the ligands, and identifying an antibody with the requisite potency against all of the potential ligands IL1-α, IL1-β, IL-33, and IL-36α, β, γ is considered a challenge, suggesting that the IL1RAP binding interface may differ between the various co-receptors. Therefore, the binding epitope appears to be crucial for efficacy, since not all antibodies have the ability to block signaling, even if they have the ability to bind to IL1RAP and mediate efficient ADCC (Agerstam, et al. (2015) PNAS 112 (34):10786-91).

[0006] The diverse nature of inflammatory diseases may explain the suboptimal performance or even failure in clinical trials when researchers simply attempt to neutralize a single disease-driving cytokine at a time, particularly in common cytokines that attempt to synergize and / or provide feedback to other cytokine members in the pathway. The IL-1 family may represent such a convergence in many inflammatory and disease conditions. IL-1β and IL-33 in asthma (Lappalainen, et al. (2005) Interleukin-1β causes pulmonary inflammation, emphysema, and airway remodeling in the adult murine lung. Am. J. Respir. Cell Mol. Biol. 32, 311-318;Prefontaine, D. et al. (2009) Increased expression of IL-33 in severe asthma: evidence of expression by airway smooth muscle cells. J. Immunol. 183, 5094-5103), and IL-1α and IL-36 in psoriasis (Towne, JE & Sims, JE (2012) IL-36 in psoriasis. Curr. Opin. Pharmacol. 12, 486-490 (2012); and Tortola, L. et al. (2012) Psoriasiform dermatitis is driven by In these diseases, more than one IL-1 family member is thought to play a prominent role, such as IL-36-mediated DC-keratinocyte crosstalk. J. Clin. Invest. 122, 3965-3976. Therefore, optimal outcomes may be expected by blocking multiple IL-1-related pathways.

[0007] Recent publications have disclosed in vitro findings that their IL1RAP antibodies specifically inhibit signaling of all six IL-1 family members (IL-1, IL-33, and IL-36).More recently, Hojen et al. published their findings of a mAb against IL1RAP that specifically inhibits IL-1, IL-33, and IL-36 signaling in vitro and also significantly attenuates heterogeneous cytokine-driven inflammation and disease severity (Hojen et al (2019) IL-1R3 blockage broadly attenuates the function of six members of the IL-1 family, revealing their contribution to models of disease. Nature Immunol. 20: 1138-1149).

[0008] IL1RAP is a co-receptor in three signaling pathways involving six cytokines of the IL-1 family (IL-1α, IL-1β, IL-33, IL36α, IL-36β, and IL-36γ), and since many diseases are driven by these cytokines, a single antagonist agent such as an antibody that can block all of the pathways would be of considerable therapeutic benefit, especially in the treatment of inflammatory diseases. Therefore, what is needed is an IL1RAP antibody that has broader simultaneous inhibition across multiple or all IL1 cytokines and more potent biological activity. Summary of the Invention

[0009] The present invention addresses the above needs by providing a biotherapeutic, particularly an antibody, that binds to IL1RAP. In one embodiment, the antibody of the present invention blocks IL1RAP-mediated cytokine signaling through the IL-33 and IL-36 signaling pathway family of key inflammatory cytokines. In one embodiment, the antibody of the present invention is useful for treating epithelial-mediated inflammation / fibrosis in diseases such as, for example, psoriasis, asthma, autoimmune diseases, acute infection scleroderma, COPD, and chronic kidney disease. In one aspect, the invention provides an anti-IL1RAP antibody having one or more of the following characteristics: In one embodiment, the anti-IL1RAPR antibody of the present invention has high molecule / cell binding potency. D In a further embodiment, the anti-IL1RAP antibodies of the invention, particularly humanized anti-IL1RAP antibodies, bind to human IL1RAP with a K D Binds to human and cynomolgus IL1RAP with <200 pM.

[0010] In embodiment 2, the present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 1, wherein said antibody or antigen-binding fragment is a monoclonal antibody or antigen-binding fragment thereof. In embodiment 3, the present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 1 or 2, wherein said antibody or antigen-binding fragment is a humanized antibody or antigen-binding fragment thereof. In embodiment 4, the present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 3, which blocks IL-1, IL-33, and IL-36 signaling. In embodiment 5, the present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, which does not bind to human IL-1R1, IL-33R, and IL-36R.

[0011] In embodiment 6, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 3, 6, 117, 118, 119, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135 (L-CDR1); the amino acid sequence of SEQ ID NO: 4, 7, 136, 137, 138, 139, 140, or 141 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8, 11, 12, 14, 142, 143, 144, 145, 146, or 147 (H-CDR1); the amino acid sequence of SEQ ID NO: 9, 13, 15, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, or 167 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 or 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 1, comprising:

[0012] In embodiment 7, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1=M or I, and X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1=M, I, X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 112 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3), or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (wherein amino acid X1=D or G, and X2=T or A) (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0013] In embodiment 8, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1=M or I, and X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1=M, I, X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 112 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3), or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 7 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (wherein amino acid X1=D or G, and X2=T or A) (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0014] In embodiment 9, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1=M or I, and X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1=M, I, X2=N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1=D or G; X2=A or T; X3=N or A; X4=Q or E; X5=M or K; X6=Q or K; X7=D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 112 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3), or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 139 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (wherein amino acid X1=D or G, and X2=T or A) (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0015] In embodiment 10, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:9 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 9 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0016] In embodiment 11, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0017] In embodiment 12, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0018] In embodiment 13, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0019] In embodiment 14, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:148 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0020] In embodiment 15, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0021] In embodiment 16, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1=A or T; X2=Q or E; X3=S or N; X4=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1=K or S; X2=S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134 (wherein amino acids X1=A or T; X2=Q or E; X3=S or N; X4=N or S); the amino acid sequence (L-CDR2) of SEQ ID NO: 140 (wherein amino acids X1=K or S; X2=S or T); and the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1=Q or E; X2=S or N; X3=N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1=K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 163 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0022] In embodiment 17, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:9 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 9 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0023] In embodiment 18, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 7 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0024] In embodiment 19, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3), or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 7 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0025] In embodiment 20, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 7 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0026] In embodiment 21, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:148 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 139 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0027] In embodiment 22, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 139 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0028] In embodiment 23, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 139 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 163 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to embodiment 6, comprising:

[0029] In embodiment 24, the invention relates to an antibody or antigen-binding fragment thereof comprising: a light chain variable region comprising the amino acid sequence of SEQ ID NO: 17; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 67; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:36; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:86; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 40; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 90; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:47; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:50; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:100; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:51; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:101; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:52; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:102. The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof according to any one of embodiments 17 to 23, comprising:

[0030] In embodiment 25, the invention provides an anti-IL1RAP antibody, wherein the antibody comprises a light chain comprising the amino acid sequence of any one of SEQ ID NOs: 170, 171, 172, 173, 174, 175, or 176; and a heavy chain comprising the amino acid sequence of any one of SEQ ID NOs: 177, 178, 179, 180, 181, 182, or 183. In embodiment 26, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 170; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 177. In embodiment 27, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 171; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 178. In embodiment 28, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 172; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 179.

[0031] In embodiment 29, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 173; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 180. In embodiment 30, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 174; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 181. In embodiment 31, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 175; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 182. In embodiment 32, the invention provides an anti-IL1RAP antibody according to embodiment 25, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 176; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 183.

[0032] In one embodiment, the antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 32 is a monoclonal antibody. In one embodiment, the antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 25 is a humanized antibody. In one embodiment, the antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 25 is a monoclonal humanized antibody.

[0033] In embodiment 33, the present invention relates to an antibody or antigen-binding fragment thereof comprising: a light chain variable region comprising an amino acid sequence of SEQ ID NO: 3, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 134 (L-CDR1); an amino acid sequence of SEQ ID NO: 4, 136, 137, 138, or 140 (L-CDR2); an amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 8, 142, 143, or 146 (H-CDR1); an amino acid sequence of SEQ ID NO: 9, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, or 165 (H-CDR2); an amino acid sequence of SEQ ID NO: 10 (H-CDR3), or a light chain variable region comprising an amino acid sequence of SEQ ID NO: 3, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 134 (L-CDR1); an amino acid sequence of SEQ ID NO: 4, 136, 137, 138, or 140 (L-CDR2); an amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 11, 144, 145, or 147 (H-CDR1); an amino acid sequence of SEQ ID NO: 9, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, or 165 (H-CDR2); an amino acid sequence of SEQ ID NO: 10 (H-CDR3), or a light chain variable region comprising an amino acid sequence of SEQ ID NO: 3, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 134 (L-CDR1); an amino acid sequence of SEQ ID NO: 4, 136, 137, 138, or 140 (L-CDR2); an amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 12, 142, 143, or 146 (H-CDR1); an amino acid sequence of SEQ ID NO: 13, 161 (H-CDR2); an amino acid sequence of SEQ ID NO: 10 (H-CDR3), or a light chain variable region comprising an amino acid sequence of SEQ ID NO: 6, 127, 128, 129, 130, 131, 132, 133, 134, or 135 (L-CDR1); an amino acid sequence of SEQ ID NO: 7, 139, or 141 (L-CDR2); an amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 14 (H-CDR1); an amino acid sequence of SEQ ID NO: 15, 162, 163, 164, or 167 (H-CDR2); an amino acid sequence of SEQ ID NO: 16 (H-CDR3). The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0034] In embodiment 34, the invention relates to an antibody or antigen-binding fragment thereof comprising: a light chain variable region comprising the amino acid sequence of SEQ ID NO: 17; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 67; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 18; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 68; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 19; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 69; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:20; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:70; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:21; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:71; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:22; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:72; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:23; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:73; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:24; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:74; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:25; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:75; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:26; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:76; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:27; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:28; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:78; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:29; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:79; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:30; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:80; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:31; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:81; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:32; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:82; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 33; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 83; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:34; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:84; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:35; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:85; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:36; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:86; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:37; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:87; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:38; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:88; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:39; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:89; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 40; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 90; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:41; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:91; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 42; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 92; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 43; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 93; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 44; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 94; or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 45; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 95; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:46; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:96; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:47; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:48; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:98; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:49; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:99; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:50; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:100; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:51; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:101; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:52; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:102; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:53; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:103; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:54; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:104; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:55; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:105; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:56; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:106; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:57; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:107; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:58; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:108; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:59; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:109; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:60; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:110; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:61; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:111; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:62; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:112; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:63; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:113; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:64; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:114; or a light chain variable region comprising the amino acid sequence of SEQ ID NO:65; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:115; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:66; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:116. The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0035] In a further embodiment 35, the invention provides a pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of the preceding embodiments, and a pharma- ceutically acceptable carrier. In a further embodiment 36, the present invention provides an antibody or antigen-binding fragment or a pharmaceutical composition according to any one of the preceding embodiments for use as a medicament. In a further embodiment 37, the invention provides the antibody or antigen-binding fragment, or pharmaceutical composition according to any one of embodiments 1 to 34, wherein the use is for the treatment of an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, or cancer.

[0036] In a further embodiment 38, the present invention provides an anti-IL1RAP antibody or antigen-binding fragment according to any one of embodiments 1 to 34 for use in treating a disease selected from an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, and cancer. In a further embodiment 39, the present invention provides the use of an anti-IL1RAP antibody or antigen-binding fragment according to any one of embodiments 1 to 34 in the manufacture of a medicament for treating a disease selected from an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, and cancer. In a further embodiment 40, the present invention provides the use of an antibody or antigen-binding fragment according to any one of embodiments 1 to 34, an anti-IL1RAP antibody or antigen-binding fragment according to embodiment 38, or an anti-IL1RAP antibody or antigen-binding fragment according to embodiment 39, wherein the use is for the treatment of psoriasis, psoriatic arthritis, multiple sclerosis, rheumatoid arthritis, COPD, chronic asthma, or ankylosing spondylitis.

[0037] In another embodiment 41, the present invention provides a method for treating a disease comprising administering to a patient in need thereof an antibody or antigen-binding fragment, or a pharmaceutical composition according to any one of embodiments 1-34, wherein the disease is selected from an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, and cancer. In another embodiment 42, the invention provides a method according to embodiment 41, wherein the disease is selected from psoriasis, psoriatic arthritis, multiple sclerosis, rheumatoid arthritis, COPD, chronic asthma, and ankylosing spondylitis.

[0038] Further embodiments of the invention include: - an isolated polynucleotide, preferably a DNA or RNA sequence, comprising a sequence encoding an anti-IL1RAP antibody or antigen-binding fragment according to the invention; - an isolated polynucleotide according to the invention, encoding a sequence defined by one or more of SEQ ID NOs: 1 to 167 or 170 to 183; - a vector comprising a polynucleotide according to the invention, preferably an expression vector, more preferably a vector comprising a polynucleotide according to the invention operatively linked to an expression control sequence; - a host cell comprising a polynucleotide according to the invention and / or a vector according to the invention; - a method for the production of an anti-IL1RAP antibody or antigen-binding fragment according to the invention, preferably a recombinant production method comprising the use of a polynucleotide according to the invention and / or of a vector according to the invention and / or of a host cell according to the invention; - such a method preferably comprises the steps of: (a) culturing a host cell under conditions allowing expression of an anti-IL1RAP antibody or antigen-binding fragment, and (b) recovering the anti-IL1RAP antibody or antigen-binding fragment; - an anti-IL1RAP antibody or antigen-binding fragment according to the invention or a diagnostic kit or method comprising its use; - a diagnostic kit or a diagnostic method according to the invention for the diagnosis of an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, cancer, psoriasis, psoriatic arthritis, multiple sclerosis, rheumatoid arthritis, COPD, chronic asthma, or ankylosing spondylitis. [Brief description of the drawings]

[0039]

Figure 1

Figure 2A-D

Figure 3A-B

Figure 4A-D

Figure 5A-D

Figure 6A-B

Figure 7A-C

[0040] Description of the Invention The present invention relates to anti-IL1RAP antibodies. In one aspect, the antibodies of the invention are for diagnostic and therapeutic use, for example in humans.

[0041] The present invention provides antibodies that bind to IL1RAP, particularly human IL1RAP. The present invention also relates to humanized antibodies that bind to IL1RAP. In a specific embodiment, the sequences of these humanized antibodies are identified based on the sequences of certain lead murine antibodies.

[0042] Without wishing to be bound by this theory, it is believed that anti-IL1RAP antibodies or antigen-binding fragments thereof bind to human IL1RAP and therefore interfere with the binding of IL1RAP agonists, thereby at least partially blocking the signaling cascade from IL1RAP to inflammatory mediators.

[0043] In one aspect, the invention provides an anti-IL1RAP antibody having one or more of the following characteristics: In one embodiment, the anti-IL1RAP antibody of the present invention has high molecule / cell binding potency. DIn a further embodiment, the anti-IL1RAP antibodies of the invention, particularly humanized anti-IL1RAP antibodies, bind to human IL1RAP with a K D Binds to human and cynomolgus IL1RAP with <200 pM. In another embodiment, the anti-IL1RAP antibodies of the invention have high cell-based functional blocking potency. In one embodiment, the anti-IL1RAP antibody of the present invention is a humanized antibody. In one embodiment, the anti-IL1RAP antibody of the present invention is a monoclonal antibody. In one embodiment, the anti-IL1RAP antibody of the present invention is a full-length antibody. In one embodiment, the anti-IL1RAP antibody of the present invention is a humanized monoclonal antibody, e.g., a full-length humanized monoclonal antibody.

[0044] The antibodies or antigen-binding fragments thereof of the present invention recognize a specific "IL1RAP antigenic epitope" or "IL1RAP epitope" or "IL-1RAP epitope." As used herein, these terms refer to a molecule (e.g., a peptide) or a fragment of a molecule capable of immunoreactivity with an anti-IL1RAP antibody. Epitopes are most commonly proteins, short oligopeptides, oligopeptide mimetics (i.e., organic compounds that mimic the antibody binding properties of the IL1RAP antigen), or combinations thereof. The minimum size of a peptide or polypeptide epitope for an antibody is believed to be about 4-5 amino acids. A peptide or polypeptide epitope contains, for example, at least 7 amino acids, or, for example, at least 9 amino acids, or, for example, about 15 to about 20 amino acids. Because an antibody may recognize an antigenic peptide or polypeptide in its tertiary form, the amino acids that comprise the epitope need not be contiguous, and in some cases, may not even be on the same peptide chain. Epitopes may be determined by a variety of techniques known in the art, such as X-ray crystallography, hydrogen / deuterium exchange mass spectrometry (HXMS), site-directed mutagenesis, alanine scanning mutagenesis, and peptide screening methods.

[0045] The generalized structure of an antibody or immunoglobulin is well known to those skilled in the art. These molecules are heterotetrameric glycoproteins, typically of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains, and are typically referred to as full-length antibodies. Each light chain is covalently linked to a heavy chain by one disulfide bond to form a heterodimer, and the heterotetrameric molecule is formed by covalent disulfide linkages between the two identical heavy chains of the heterodimer. The light and heavy chains are linked together by one disulfide bond, while the number of disulfide linkages between the two heavy chains varies with the immunoglobulin isotype. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain contains a variable domain (VH) at the amino terminus. H ), followed by three or four constant domains (C H1 , C H2 , C H3 , and C H4 ), and C H1 and C. H2 Each light chain has two domains, an amino-terminal variable domain (V L ) and the carboxy-terminal constant domain (C L ) V L Domain is V H domain and non-covalently binds to C L The domains are connected via disulfide bonds to C H1 The domains are generally covalently linked. Certain amino acid residues are believed to form an interface between the light and heavy chain variable domains (Chothia et al., 1985, J. Mol. Biol. 186:651-663). The variable domain is also referred to herein as the variable region.

[0046] Certain domains within the variable domains vary widely among different antibodies, i.e., are "hypervariable". These hypervariable domains contain the residues that are directly involved in the binding and specificity of each particular antibody to its specific antigenic determinant. Hypervariability is concentrated in three segments known as complementarity determining regions (CDRs) or hypervariable loops (HVLs) in both the light and heavy chain variable domains. CDRs are described by Kabat et al., 1991, In: Sequences of Proteins of Immunological Interest, 5 thThe CDRs are defined by sequence comparison in the Ed. Public Health Service, National Institutes of Health, Bethesda, Md., whereas the HVLs (also referred to herein as CDRs) are structurally defined according to the three-dimensional structure of the variable domain as described by Chothia and Lesk, 1987, J. Mol. Biol. 196: 901-917. These two methods result in slightly different identifications of the CDRs. As defined by Kabat, in the light chain variable domain, CDR-L1 is located at about residues 24-34, CDR-L2 at about residues 50-56, and CDR-L3 at about residues 89-97; in the heavy chain variable domain, CDR-H1 is located at about residues 31-35, CDR-H2 at about residues 50-65, and CDR-H3 at about residues 95-102. The exact residue numbers encompassing a particular CDR will vary depending on the sequence and size of the CDR. While the Kabat and Chothia scheme treats different families of immunoglobulin domains separately, Lefranc and colleagues proposed a unified numbering scheme, referred to as the IMGT numbering scheme, for immunoglobulin variable domain genomic sequences, including Ab light and heavy variable domains, and T cell receptor variable domains (Lefranc MP, et al. IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains and Ig superfamily V-like domains. Dev Comp Immunol (2003) 27:55-77).

[0047] As mentioned above, there are differences and shortcomings among the Kabat, Chothia, and IMGT numbering schemes, and the variable CDR lengths only take into account the most frequent loop lengths, and therefore there are some discrepancies depending on the numbering scheme used. In some cases, some of the CDRs identified by the four methods included herein are nearly identical (e.g., L3, H3), while other CDRs (e.g., L2, H1, and H2) have substantial differences between the methods. Therefore, for completeness, the CDRs of this embodiment are labeled according to each of the numbering conventions that best identify the critical Ag-binding residues (see Tables 1-4 below). One of skill in the art can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of an antibody. Thus, the CDR1, CDR2, and CDR3 of the heavy and light chains define the specific, intrinsic, and functional properties of a given antibody.

[0048] CDR sequences according to four different nomenclature systems CDRs 1-3 of the variable light and variable heavy chain regions for antibodies A#1-A#7 are listed by SEQ ID NO: in Table 1 below and presented according to Kabat nomenclature.

[0049] [Table 1-1] [Table 1-2]

[0050] CDRs 1-3 of the variable light and variable heavy chain regions for antibodies A#1-A#7 are presented in Table 2 below according to the CCG (Chemical Computing Group, as exemplified in Almagro et al., Proteins 2011; 79:3050-3066 and Maier et al, Proteins 2014; 82:1599-1610).

[0051] [Table 2-1] [Table 2-2]

[0052] An additional numbering system based on the Chothia scheme is presented for CDRs 1-3 of the variable light and variable heavy chain regions for antibodies A#1-A#7 in Table 3 below.

[0053] [Table 3-1] [Table 3-2]

[0054] CDRs 1-3 of the variable light and variable heavy chain regions for antibodies A#1-A#7C based on the IMGT numbering scheme (Lefranc MP, et al. IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains and Ig superfamily V-like domains. Dev Comp Immunol (2003) 27:55-77) are presented in Table 4 below.

[0055] [Table 4-1] [Table 4-2]

[0056] The amino acid positions denoted in this context for Kabat, CCG, Chothia, or IMGT positions (Tables 1-4) are linear, i.e., the amino acids of each full length molecular chain are numbered consecutively starting with number 1 at the N-terminus and ending with a number that corresponds to the total number of amino acids in the molecule. For example, a heavy chain 118 amino acids in length would start with number 1 at the N-terminus and end with number 118 at the most C-terminal amino acid. Thus, for example, any reference to position 25 means amino acid number 25 counting from the N-terminus of the molecule.

[0057] The three CDRs in each of the heavy and light chains are separated by framework regions (FRs) that contain sequences that tend to be less variable. From the amino terminus to the carboxy terminus of the heavy and light chain variable domains, the FRs and CDRs are arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The predominantly β-sheet conformation of the FRs brings the CDRs in each of the chains into close proximity with each other and with the CDRs from the other chain. The resulting conformation contributes to the antigen-binding site (see Kabat et al., 1991, NIH Publ. No. 91-3242, Vol. I, pages 647-669), although not all CDR residues are directly involved in antigen binding.

[0058] FR residues and Ig constant domains are not directly involved in antigen binding, but contribute to antigen binding and / or mediate antibody effector functions. Some FR residues are thought to have significant effects on antigen binding in at least three ways: by directly binding non-covalently to the epitope, by interacting with one or more CDR residues, and by influencing the interface between the heavy and light chains. Constant domains are not directly involved in antigen binding, but mediate various Ig effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and antibody-dependent cellular phagocytosis (ADCP).

[0059] The light chains of vertebrate immunoglobulins are assigned to one of two clearly distinct classes, kappa (κ) and lambda (λ), based on the amino acid sequence of the constant domain. In comparison, the heavy chains of mammalian immunoglobulins are assigned to one of five major classes: IgA, IgD, IgE, IgG, and IgM, according to the sequence of their constant domains. IgG and IgA are further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains that correspond to the various classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the classes of native immunoglobulins are well known.

[0060] The terms "antibody", "anti-IL1RAP antibody", "anti-IL1-RAP antibody", "humanized anti-IL1RAP antibody", "humanized anti-IL1RAP epitope antibody", and "variant humanized anti-IL1RAP epitope antibody" specifically encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments such as variable domains and other portions of antibodies that exhibit the desired biological activity, e.g., IL1RAP binding. Humanized antibodies are largely human immunoglobulins (e.g., chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin) in which residues from the recipient complementarity determining regions (CDRs) are replaced by residues from the CDRs of a non-human species, such as mouse, rat, or rabbit, that have the desired specificity, affinity, and biological activity. The term "monoclonal antibody" (mAb) refers to an antibody that is highly specific to a single antigenic determinant "epitope." Thus, the modifier "monoclonal" refers to antibodies to the same epitope and should not be construed as requiring production of the antibody by any particular method. It should be understood that monoclonal antibodies can be made by any technique or methodology known in the art, including, for example, the hybridoma method (Kohler et al., 1975, Nature 256:495), or recombinant DNA methods known in the art (see, e.g., U.S. Pat. No. 4,816,567), or methods for isolating recombinantly produced monoclonals using phage antibody libraries, using the techniques described in Clackson et al., 1991, Nature 352: 624-628 and Marks et al., 1991, J. Mol. Biol. 222: 581-597. The term "monomeric" refers to a homogeneous form of an antibody. For example, with respect to a full length antibody, monomer means a monomeric antibody having two identical heavy chains and two identical light chains.

[0061] A chimeric antibody consists of the heavy and light chain variable regions of an antibody from one species (e.g., a non-human mammal such as a mouse) and the heavy and light chain constant regions of an antibody from another species (e.g., human), and can be obtained by linking DNA sequences encoding the variable regions of an antibody from a first species (e.g., a mouse) to DNA sequences for the constant regions of an antibody from a second species (e.g., human) and transforming a host with an expression vector containing the linked sequences to cause it to produce the chimeric antibody. Alternatively, a chimeric antibody can be one in which one or more regions or domains of the heavy and / or light chain are identical to, homologous to, or variants of corresponding sequences in a monoclonal antibody from another immunoglobulin class or isotype, or from a consensus or germline sequence. Chimeric antibodies can include fragments of such antibodies, provided that the antibody fragment exhibits the desired biological activity of the parent antibody, e.g., binding to the same epitope (see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al., 1984, Proc. Natl. Acad. Sci. USA 81: 6851-6855).

[0062] The terms "antibody fragment", "anti-IL1RAP antibody fragment", "anti-IL1RAP epitope antibody fragment", "humanized anti-IL1RAP antibody fragment", "humanized anti-IL1RAP epitope antibody fragment", "variant humanized anti-IL1RAP epitope antibody fragment" refer to a portion of a full-length anti-IL1RAP antibody that retains the variable region or functional capability, such as specific IL1RAP epitope binding. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fd, Fv, scFv, and scFv-Fc fragments, diabodies, linear antibodies, single chain antibodies, minibodies, diabodies formed from antibody fragments, and multispecific antibodies formed from antibody fragments.

[0063] Full length antibodies can be treated with enzymes such as papain or pepsin to produce useful antibody fragments. Papain digestion is used to produce two identical antigen-binding antibody fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment. The Fab fragment contains the constant domains of the light chain and the C domain of the heavy chain. H1 Pepsin treatment yields an F(ab')2 fragment that has two antigen-binding sites and is still capable of cross-linking antigen. Fab' fragments are H1 They differ from Fab fragments by the presence of additional residues at the C-terminus of the domain, including one or more cysteines from the antibody hinge region. F(ab')2 antibody fragments are pairs of Fab' fragments linked by cysteine ​​residues in the hinge region. Other chemical couplings of antibody fragments are also known.

[0064] An "Fv" fragment contains a complete antigen recognition and binding site, consisting of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. In this configuration, the three CDRs of each variable domain interact to form the V H -V L Collectively, the six CDRs define an antigen-binding site on the surface of the dimer and confer antigen-binding specificity to the antibody. "Single-chain Fv" or "scFv" antibody fragments are fragments of the V H and V L A single chain Fv variant contains a V domain, where the domain is present in a single polypeptide chain. A single chain Fv can recognize and bind to an antigen. An scFv polypeptide is a single chain Fv variant that contains a V domain, where the domain is present in a single polypeptide chain. H and V LA polypeptide linker located between the domains can also be optionally included to facilitate formation of the desired three-dimensional structure for antigen binding by the scFv (see, e.g., Pluckthun, 1994, In The Pharmacology of monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315).

[0065] "Diabody" refers to a small antibody fragment having two antigen binding sites, which fragment comprises a heavy chain variable domain (V.sub.H) connected to a light chain variable domain (V.sub.L) in the same polypeptide chain (V.sub.HV.sub.L or V.sub.LV.sub.H). Diabodies are more fully described, for example, in Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448. Other recognized antibody fragments consist of a pair of tandem Fd segments (V H -C H1 -V H -C H1 ). These "linear antibodies" can be bispecific or monospecific, e.g., as described in Zapata et al. 1995, Protein Eng. 8(10):1057-1062.

[0066] A "humanized antibody" or "humanized antibody fragment" is a specific type of chimeric antibody that contains an immunoglobulin amino acid sequence variant or fragment thereof that can bind to a given antigen and that contains one or more FRs that have substantially the amino acid sequence of a human immunoglobulin and one or more CDRs that have substantially the amino acid sequence of a non-human immunoglobulin. This non-human amino acid sequence, often referred to as an "import" sequence, is typically selected from the "import" antibody domains, particularly the variable domains. Generally, a humanized antibody contains at least the CDRs or HVLs of a non-human antibody inserted between the FRs of a human heavy or light chain variable domain. The present invention describes a specific humanized anti-IL1RAP antibody that contains CDRs or humanized CDRs derived from a mouse monoclonal antibody inserted between the FRs of human germline sequence heavy and light chain variable domains. It will be understood that certain mouse FR residues may be important for the function of the humanized antibody, and therefore certain human germline sequence heavy and light chain variable domain residues are modified to be the same as those of the corresponding mouse sequences.

[0067] In another embodiment, the humanized anti-IL1RAP antibody comprises substantially all of at least one, typically two, variable domains (e.g., as contained in Fab, Fab', F(ab')2, Fabc, and Fv fragments), with all or substantially all of the CDRs corresponding to those of a non-human immunoglobulin, specifically herein all of the CDRs being murine or humanized sequences as detailed herein below, and all or substantially all of the FRs being of human immunoglobulin consensus or germline sequences. In another embodiment, the humanized anti-IL1RAP antibody also comprises at least a portion of an immunoglobulin Fc region, typically that of a human immunoglobulin. Typically, the antibody will contain both a light chain, as well as at least the variable domain of a heavy chain. The antibody may optionally comprise a CDR of the heavy chain, or a CDR of the human immunoglobulin consensus or germline sequence. H1 , Hinge, C H2 , C H3 , and / or C. H4 It may also include one or more of the regions.

[0068] The humanized anti-IL1RAP antibody may be selected from any class of immunoglobulin, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. For example, the constant domain may be a complement fixing constant domain, and it is desirable for the humanized antibody to exhibit cytotoxic activity, with the isotype typically being IgG1. If such cytotoxic activity is not desired, the constant domain may be of another isotype, for example IgG2. Alternative humanized anti-IL1RAP antibodies may comprise sequences from more than one immunoglobulin class or isotype, and it is within the ordinary skill in the art to select a particular constant domain to optimize desired effector functions. In a specific embodiment, the invention provides an antibody that is an IgG1 antibody, more particularly an IgG1 antibody with knockout of effector function.

[0069] The FRs and CDRs or HVLs of the humanized anti-IL1RAP antibody need not correspond exactly to the parental sequences. For example, one or more residues in the import CDRs or HVLs, or the consensus or germline FR sequences, can be altered (e.g., mutagenized) by substitution, insertion, or deletion such that the resulting amino acid residue is no longer identical to the original residue at the corresponding position in either parental sequence, but the antibody nevertheless retains the function of binding to IL1RAP. Such alterations will typically not be drastic, but will be conservative changes. Usually, at least 75%, more often at least 90%, and most frequently more than 95% or more than 98% or more than 99% of the humanized antibody residues will correspond to those of the parental consensus or germline FR and import CDR sequences. The interface between the heavy and light chain variable regions ("V L -V H Immunoglobulin residues that affect the "interface" are those that affect the proximity or orientation of the two chains with respect to each other. Certain residues that may be involved in interchain interactions are V LResidues 34, 36, 38, 44, 46, 87, 89, 91, 96, and 98, and V H It contains residues 35, 37, 39, 45, 47, 91, 93, 95, 100, and 103 (using the numbering system set forth in Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md., 1987)). L Residues 43 and 85, and V H It is argued that residues such as residues 43 and 60 may also be involved in this interaction. Although these residues are only represented for human IgG, they are applicable across species. Key antibody residues that are reasonably predicted to be involved in interchain interactions are selected for substitution into the consensus sequence.

[0070] The terms "consensus sequence" and "consensus antibody" refer to an amino acid sequence that contains the most frequently occurring amino acid residue at each position in all immunoglobulins of any particular class, isotype, or subunit structure, such as human immunoglobulin variable domains. A consensus sequence may be based on immunoglobulins of a particular species or of many species. A "consensus" sequence, structure, or antibody is understood to refer to an amino acid sequence that contains the most frequently occurring amino acid residue at each position in all human immunoglobulins of any particular class, isotype, or subunit structure, including the consensus human sequence described in certain embodiments. Thus, a consensus sequence contains an amino acid sequence that has amino acids at each position that are present in one or more known immunoglobulins, but may not exactly reproduce the entire amino acid sequence of any single immunoglobulin. The variable region consensus sequences are not obtained from any naturally occurring antibody or immunoglobulin, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., and variants thereof.

[0071] Human germline sequences are naturally found in the human population. The combination of these germline genes creates antibody diversity. Germline antibody sequences for antibody light chains arise from conserved human germline kappa or lambda v and j genes. Similarly, heavy chain sequences arise from germline v, d, and j genes (LeFranc, MP, and LeFranc, G, "The Immunoglobulin Facts Book" Academic Press, 2001). As used herein, "variant," "anti-IL1RAP variant," "humanized anti-IL1RAP variant," or "variant humanized anti-IL1RAP" each refer to a humanized anti-IL1RAP antibody having at least the light chain variable murine CDRs. Variants include those with one or more amino acid changes in one or both light or heavy chain variable domains, provided that the amino acid changes do not substantially impair binding of the antibody to IL1RAP.

[0072] An "isolated" antibody is one that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of the antibody's natural environment are such materials that would interfere with diagnostic or therapeutic uses for the antibody, and may be enzymes, hormones, or other proteinaceous or nonproteinaceous solutes. In one embodiment, the antibody will be purified to isolate at least greater than 95% by weight of the antibody. Isolated antibody includes the antibody in situ within recombinant cells in which it is produced, since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step in which recombinant cell material is removed.

[0073] The term "antibody performance" refers to factors that contribute to the antibody's recognition of an antigen or the effectiveness of an antibody in vivo. Changes in the amino acid sequence of an antibody can affect antibody properties such as folding and the initial rate of antibody binding to an antigen (k a ), the dissociation constant of the antibody from the antigen (k d ), the affinity constant (Kd) of the antibody for the antigen, the three-dimensional structure of the antibody, protein stability, and physical factors such as the half-life of the antibody.

[0074] The term "epitope tagged" as used herein refers to an anti-IL1RAP antibody fused to an "epitope tag." An "epitope tag" is a polypeptide having a sufficient number of amino acids to provide an epitope for antibody production, yet is designed such that it does not interfere with a desired activity of the humanized anti-IL1RAP antibody. The epitope tag is typically sufficiently unique so that an antibody made against the epitope tag does not substantially cross-react with other epitopes. Suitable tag polypeptides generally contain at least six amino acid residues, and typically contain about 8-50 amino acid residues, or about 9-30 residues. Examples of epitope tags and antibodies that bind to the epitopes include the flu HA tag polypeptide and its antibody 12CA5 (Field et al., 1988 Mol. Cell. Biol. 8: 2159-2165); c-myc tag and its 8F9, 3C7, 6E10, G4, B7, and 9E10 antibodies (Evan et al., 1985, Mol. Cell. Biol. 5(12):3610-3616); and herpes simplex virus glycoprotein D (gD) tag and its antibodies (Paborsky et al. 1990, Protein Engineering 3(6): 547-553). In certain embodiments, the epitope tag is a "salvage receptor binding epitope." As used herein, the term "salvage receptor binding epitope" refers to an epitope in the Fc region of an IgG molecule (such as IgG1, IgG2, IgG3, or IgG4) that is responsible for increasing the in vivo serum half-life of the IgG molecule.

[0075] In some embodiments, the antibodies of the invention may be conjugated to a cytotoxic agent, which is any substance that inhibits or prevents the function of cells and / or causes destruction of cells. The term includes radioisotopes (I 131 , I 125 , Y 90 , and Re 186Cytotoxic agents are intended to include cytotoxic agents, such as cytotoxic agents, chemotherapeutic agents, and toxins, such as enzymatically active toxins of bacterial, fungal, plant, or animal origin, and fragments thereof. Such cytotoxic agents can be conjugated to the humanized antibodies of the invention using standard procedures and used, for example, to treat patients eligible for antibody-based therapy.

[0076] A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. There are numerous examples of chemotherapeutic agents that may be conjugated to the therapeutic antibodies of the invention. Examples of such chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine; acetogenins (especially bullatacin and bullatasinone); camptothecins, and the like. statins (including the synthetic analog topotecan); bryostatins; kallistatins; CC-1065 (including its adozelesin, carzelesin, and bizelesin synthetic analogs); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatins, auristatins (including the analogs monomethyl-auristatin E and monomethyl-auristatin F); duocarmycins (including the synthetic analogs KW-2189 and CBI-TMI); eleutherobin; pancratistatins; sarcodictiins; spongistatins; chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride oxide oxide hydrochloride), nitrogen mustards such as melphalan, novembichine, phenesterine, and prednimustine; trofosfamide and uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine;Enediyne antibiotics (e.g., the calichemicins, especially calichemicin gamma 1I and calichemicin phiI1, see, e.g., Agnew, Chem. Intl. Ed. Engl., 33:183-186; the dynemicins, including dynemicin A; the bisphosphonates, such as clodronate; the esperamicins; and the neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (Adriamycin™) (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin Antibiotics such as mitomycins such as marcelomycin and mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; metabolic antagonists such as methotrexate and 5-fluorouracil (5-FU). substances; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate, etc.; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, etc.; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, etc.; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, etc.;Anti-adrenal drugs such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as frolinic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; democolcine; diazicon; elfomithine; elliptinium acetate; epothilone; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone, mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; rosoxantrone; podof diethyl phosphate; 2-ethylhydrazide; procarbazine; PSK®; razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triazicon; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethane; vindesine; dacarbazine; mannomustine; mitabronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ), and doxetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine (Gemzar™); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine (Navelbine™); novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11;Topoisomerase inhibitors include RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharma- ceutically acceptable salts, acids, or derivatives of any of the above. Antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including Nolvadex™), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxyphene, keoxifene, LY117018, onapristone, and toremifene (Fareston™); for example, 4(5)-imidazole, aminoglutethimide, megestrol acetate (Megace™), exemestane, formestane, fadro. Also included in this definition are aromatase inhibitors, which inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as aztreonam, vorozole (Rivisor™), letrozole (Femara™), and anastrozole (Arimidex™); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and antihormonal agents, which act to regulate or inhibit hormone action on tumors, such as pharmacologic acceptable salts, acids, or derivatives of any of the above. Any one or more of these agents may be conjugated to the humanized antibodies of the invention to provide useful therapeutic agents for the treatment of a variety of disorders.

[0077] Antibodies can also be conjugated to prodrugs. A "prodrug" is a precursor or derivative form of a pharma- ceutical active substance that can be enzymatically activated or converted to a more active form that is less cytotoxic to tumor cells compared to the parent drug. See, for example, Wilman, 1986, "Prodrugs in Cancer Chemotherapy", In Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Belfast, and Stella et al., 1985, "Prodrugs: A Chemical Approach to Targeted Drug Delivery, In: "Directed Drug Delivery, Borchardt et al., (ed.), pp. 247-267, Humana Press. Useful prodrugs include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs, and optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine, and other 5-fluorouridine prodrugs that can be converted to more active cytotoxic free drugs. Examples of cytotoxic drugs that can be derivatized into prodrug forms include, but are not limited to, those chemotherapeutic agents described above.

[0078] For diagnostic and therapeutic monitoring purposes, the antibodies of the present invention can also be conjugated to a label, whether it is a label alone or a label and an additional second agent (prodrug, chemotherapeutic agent, etc.). A label, as distinguished from other second agents, refers to an agent that is a detectable compound or composition, which can be directly or indirectly conjugated to the humanized antibody of the present invention. The label can itself be detectable (e.g., a radioactive or fluorescent label) or, in the case of an enzymatic label, can catalyze a chemical alteration of a substrate compound or composition that is detectable. Labeled humanized anti-IL1RAP antibodies can be prepared and used in a variety of applications, including in vitro and in vivo diagnostic methods.

[0079] The antibody of the present invention may be formulated as part of a liposomal preparation to affect its delivery in vivo. A "liposome" is a small vesicle composed of various types of lipids, phospholipids, and / or surfactants. Liposomes are useful for delivery of compounds or formulations, such as the humanized anti-IL1RAP antibodies disclosed herein, optionally conjugated to or in combination with one or more pharmacoactive agents and / or labels, to mammals. The components of the liposome are generally organized in a bilayer formation similar to the lipid organization of biological membranes.

[0080] Certain aspects of the invention relate to isolated nucleic acids encoding one or more domains of the humanized antibody of the invention. An "isolated" nucleic acid molecule is one that has been identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the antibody nucleic acid. An isolated nucleic acid molecule is distinguished from the nucleic acid molecule as it exists in natural cells. In various embodiments of the invention, one or more domains of the humanized antibody will be recombinantly expressed. Such recombinant expression may employ one or more control sequences, i.e., polynucleotide sequences necessary for the expression of an operably linked coding sequence in a particular host organism. Control sequences suitable for use in prokaryotic cells include, for example, promoters, operators, and ribosome binding site sequences. Eukaryotic control sequences include, but are not limited to, promoters, polyadenylation signals, and enhancers. These control sequences may be utilized for the expression and production of humanized anti-IL1RAP antibodies in prokaryotic and eukaryotic host cells.

[0081] A nucleic acid sequence is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, a nucleic acid presequence or secretory leader is operably linked to a nucleic acid encoding a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, "operably linked" means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading frame. However, enhancers may be contiguous. Linking can be accomplished by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adaptors or linkers can be used.

[0082] As used herein, the expressions "cell," "cell line," and "cell culture" are used interchangeably and all such designations include the progeny thereof. Thus, "transformants" and "transformed cells" include the primary subject cell and cultures derived therefrom without regard for the number of transfers. The term "mammal" for purposes of treatment refers to any animal classified as a mammal, including humans, domesticated and farm animals, and zoo, sport, or pet animals such as dogs, horses, cats, cows, etc. Preferably, the mammal is a human. A "disorder," as used herein, is any condition that would benefit from treatment with a humanized anti-IL1RAP antibody described herein. This includes chronic and acute disorders or diseases, including those conditions that predispose a mammal to the disorder in question. Non-limiting examples of disorders that are of interest to be treated herein include inflammatory, angiogenic, autoimmune, and immunological disorders, respiratory disorders, cancer, hematological malignancies, benign and malignant tumors, leukemia, and lymphoid malignancies.

[0083] The terms "cancer" and "cancerous" refer to or describe the physiological disease condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. IL1RAP-associated disorders include diseases and disorders of the immune system, such as autoimmune and inflammatory disorders, including, but not limited to, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), scleroderma, Sjogren's syndrome, multiple sclerosis, psoriasis, psoriatic arthritis, pulmonary inflammation, asthma, idiopathic thrombocytopenic purara (ITP), epithelial inflammatory disorders, fibrosis, and ankylosing spondylitis.

[0084] The term "intravenous infusion" refers to the introduction of an agent into the vein of an animal or human patient over a period of greater than about 15 minutes, generally about 30-90 minutes. The term "intravenous bolus" or "intravenous push" refers to administration of a drug into a vein of an animal or human such that the body receives the drug within approximately less than 15 minutes, typically less than 5 minutes.

[0085] The term "subcutaneous administration" refers to the introduction of an agent under the skin of an animal or human patient, preferably into a pocket between the skin and the underlying tissue, by relatively slow, sustained delivery from a drug receptacle. Pinching or lifting the skin and away from the underlying tissue can create the pocket. The term "subcutaneous infusion" refers to the introduction of a drug under the skin of an animal or human patient, preferably into a pocket between the skin and the underlying tissue, by relatively slow, sustained delivery from a drug receptacle for a period of time, including but not limited to less than 30 minutes or less than 90 minutes. Optionally, the infusion can be performed by subcutaneous implantation of a drug delivery pump implanted under the skin of the animal or human patient, the pump delivering a predetermined amount of drug for a predetermined period of time, such as 30 minutes, 90 minutes, or a period spanning the length of a treatment regimen. The term "subcutaneous bolus" refers to drug administration beneath the skin of an animal or human patient, where the bolus drug delivery is for less than approximately 15 minutes; in another embodiment, less than 5 minutes, and in yet another embodiment, less than 60 seconds. In still yet another embodiment, administration is within a pocket between the skin and the underlying tissue, where the pocket may be created by pinching or lifting the skin and away from the underlying tissue.

[0086] The term "therapeutically effective amount" is used to refer to an amount of active agent that relieves or improves one or more symptoms of the disorder being treated. In another embodiment, a therapeutically effective amount refers to a target serum concentration that has been shown to be effective, for example, in slowing down disease progression. Efficacy can be measured in a conventional manner depending on the condition being treated. The terms "treatment" and "therapy" and equivalents, as used herein, are intended to include therapeutic as well as preventive or suppressive measures against a disease or disorder that result in any clinically desirable or beneficial effect, including, but not limited to, alleviation or relief of one or more symptoms of the disease or disorder, regressing, slowing, or stopping its progression. Thus, for example, the term treatment includes administration of an agent before or after the onset of symptoms of the disease or disorder, thereby preventing or eliminating one or more signs of the disease or disorder. As another example, the term includes administration of an agent after clinical manifestation of the disease to combat symptoms of the disease. Furthermore, administration of an agent after onset and after clinical symptoms have developed includes "treatment" or "therapy" as used herein, when administration affects clinical parameters of the disease or disorder, such as the extent of tissue damage or the amount or degree of metastasis, regardless of whether the treatment leads to improvement of the disease. Furthermore, so long as a composition of the invention, alone or in combination with another therapeutic agent, alleviates or ameliorates at least one symptom of the disorder being treated, compared to that symptom without the use of a humanized anti-IL1RAP antibody composition, the result should be considered an effective treatment of the underlying disorder, regardless of whether all symptoms of the disorder have been alleviated. The term "package insert" is used to refer to instructions customarily included in commercial packaging of therapeutic products that contain information about the indications, usage, administration, contraindications, and / or warnings regarding the use of such therapeutic product.

[0087] antibody In one aspect, described and disclosed herein are anti-IL1RAP antibodies, particularly humanized anti-IL1RAP antibodies, as well as compositions and articles of manufacture comprising one or more anti-IL1RAP antibodies, particularly one or more humanized anti-IL1RAP antibodies of the invention. The variable regions and CDRs of representative antibodies of the invention are disclosed below.

[0088] Anti-IL1RAP mouse antibody sequence The variable regions and CDRs of a representative murine lead antibody (murine lead) of the invention are shown below.

[0089] Light chain variable region (VK) amino acid sequence [Table 5]

[0090] Heavy chain variable region (VH) amino acid sequence [Table 6]

[0091] Light chain CDR-1, CDR-2, CDR3 (L-CDR1-3) and heavy chain CDR-1, CDR-2, CDR3 (L-CDR1-3) amino acid sequences according to Kabat, CCG, Chothia, and IMGT nomenclature [Table 7]

[0092] [Table 8]

[0093] [Table 9]

[0094] [Table 10]

[0095] Anti-IL1RAP humanized antibody sequence Human framework sequences were selected against the murine leads based on framework homology, CDR structure, conserved canonical residues, conserved interface packing residues, and other parameters to generate humanized variable regions (see Examples 1-2).

[0096] Representative humanized variable regions derived from antibodies opt-27 through opt73 are shown below. [Table 11-1] [Table 11-2] [Table 11-3]

[0097] [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4]

[0098] Depicted below are the VL / VH from clone OPT27-73 from murine antibody 005-GO11, including the CDR sequences from the humanized variable regions, Opt-43 (#A2), Opt47 (#A3), Opt-54 (#A4), Opt-57 (#A5), Opt-58 (#A6), and Opt-59 (#A7). The CDRs are labeled according to the naming conventions KABAT, CCG, CHOTHIA, and IMGT.

[0099] [Table 13-1] [Table 13-2]

[0100] [Table 14-1] [Table 14-2]

[0101]

Table 15-1

Table 15-2

[0102]

Table 16-1

Table 16-2

[0103]

Table 17-1

Table 17-2

[0104]

Table 18-1

Table 18-2

[0105]

Table 19-1

Table 19-2

[0106]

Table 20-1

Table 20-2

[0107]

Table 21-1

Table 21-2

[0108]

Table 22-1

Table 22-2

Table 23-1

Table 23-2

[0109]

Table 24-1

Table 24-2

[0110]

Table 25-1

Table 25-2

Table 25-3

Table 25-4

Table 26-1

Table 26-2

Table 26-3

[0111] [Table 27]

[0112] Representative light and heavy chain sequences of the invention are shown below (humanized variable regions derived from the 005-GO11 parent (A1) linked to constant regions: opt-43 (A2), opt-47 (A3), opt-54 (A4), opt-57 (A5), opt-58 (A6), and opt-59 (A7)).

[0113] Light chain amino acid sequence [Table 28]

[0114] Heavy chain amino acid sequence [Table 29-1] [Table 29-2]

[0115] In one embodiment, an antibody of the invention comprises three light chain CDRs and three heavy chain CDRs, eg, as set out in the table above. In one embodiment, an antibody of the invention comprises a light chain and a heavy chain variable region as specified above. In one embodiment, a light chain variable region of the invention is fused to a light chain constant region, such as a kappa or lambda constant region. In one embodiment, a heavy chain variable region of the invention is fused to a heavy chain constant region, such as an IgA, IgD, IgE, IgG, or IgM, in particular an IgG1, IgG2, IgG3, or IgG4.

[0116] The present invention provides an anti-IL1RAP antibody (Antibody A1) comprising: a light chain comprising the amino acid sequence of SEQ ID NO:170; and a heavy chain comprising the amino acid sequence of SEQ ID NO:177. The present invention provides an anti-IL1RAP antibody (Antibody A2) comprising: a light chain comprising the amino acid sequence of SEQ ID NO:171; and a heavy chain comprising the amino acid sequence of SEQ ID NO:178. The present invention provides an anti-IL1RAP antibody (Antibody A3) comprising: a light chain comprising the amino acid sequence of SEQ ID NO: 172; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 179. The present invention provides an anti-IL1RAP antibody (Antibody A4) comprising: a light chain comprising the amino acid sequence of SEQ ID NO: 173; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 180.

[0117] The present invention provides an anti-IL1RAP antibody (Antibody A5) comprising: a light chain comprising the amino acid sequence of SEQ ID NO: 174; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 181. The present invention provides an anti-IL1RAP antibody (Antibody A6) comprising: a light chain comprising the amino acid sequence of SEQ ID NO: 175; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 182. The present invention provides an anti-IL1RAP antibody (Antibody A7) comprising: a light chain comprising the amino acid sequence of SEQ ID NO: 176; and a heavy chain comprising the amino acid sequence of SEQ ID NO: 183. Representative antibodies of the present invention are shown below.

[0118] [Table 30-1] [Table 30-2] [Table 30-3]

[0119] In some aspects, the invention includes a neutralizing antibody that binds to IL1RAP. In some embodiments, the antibody specifically binds to IL1RAP. In some embodiments, the antibody binds to domain 3 comprising amino acids 235-367 of IL1RAP (SEQ ID NO: 187). In some embodiments, the antibody binds to an epitope within amino acid residues corresponding to positions 235-315 of IL1RAP extracellular domain (ECD) 3 (SEQ ID NO: 188). In some embodiments, the antibody binds to an IL1RAP having an amino acid sequence that is at least 90% identical to SEQ ID NO: 187.

[0120] In some aspects, the invention includes an antigen binding protein that binds to IL1RAP, where the antigen binding protein binds to IL1RAP at a location within residues corresponding to positions 235-315 of SEQ ID NO: 188. In some embodiments, when the antigen binding protein binds to IL1RAP, the antibody is positioned no more than 8 Angstroms from at least one of the following residues 238, 239, 244-247, 249, 251-256, 261, 263, 265, 267, 269, 271, 301, 303, 305-306, 311, 313, or 315 of IL1RAP.

[0121] In some aspects, the invention includes an antigen binding protein that binds to IL1RAP, where the antigen binding protein binds to IL1RAP at a location within residues corresponding to positions 235-273 of SEQ ID NO: 189. In some embodiments, when the antigen binding protein binds to IL1RAP, the antibody is positioned no more than 8 Angstroms from at least one of the following residues 238, 239, 244-247, 249, 251-256, 261, 263, 265, 267, 269, or 271 of IL1RAP.

[0122] In some aspects, the invention includes an antigen binding protein that binds to IL1RAP, where the antigen binding protein binds to IL1RAP at a location within residues corresponding to positions 300-315 of SEQ ID NO: 190. In some embodiments, when the antigen binding protein binds to IL1RAP, the antibody is positioned no more than 8 Angstroms from at least one of the following residues 301, 303, 305-306, 311, 313, or 315 of IL1RAP. In some aspects, the invention includes an antigen binding protein that binds to IL1RAP, where the antigen binding protein binds to IL1RAP at a location within residues corresponding to positions 226-262 of SEQ ID NO: 192. In some embodiments, when the antigen binding protein binds to IL1RAP, the antibody is positioned no more than 8 Angstroms from at least one of the following residues 238, 239, 244-247, 249, 251-256, or 261 of IL1RAP. In some aspects, the invention includes an antigen binding protein that binds to IL1RAP, where the antigen binding protein binds to IL1RAP at a location within residues corresponding to positions 226-273 of SEQ ID NO: 192. In some embodiments, when the antigen binding protein binds to IL1RAP, the antibody is positioned no more than 8 Angstroms from at least one of the following residues 238, 239, 244-247, 249, 251-256, 261, 263, 265, 267, 269, or 271 of IL1RAP.

[0123] The antibodies of the present invention are useful in methods for the treatment of various diseases or disorders, such as immunological, inflammatory, autoimmune, and respiratory diseases in humans. For example, the antibodies of the present invention are useful in methods for the treatment of psoriasis, rheumatoid arthritis, or psoriatic arthritis. For example, the antibodies of the present invention are useful in methods for the treatment of chronic obstructive pulmonary disorder (COPD) or asthma. For example, the antibodies of the present invention are useful in methods for the treatment of scleroderma, palmoplantar pustulosis, generalized pustular psoriasis, diabetic nephropathy, lupus nephritis, scleroderma, ankylosing spondylitis, IL-36 receptor antagonist deficiency autoimmune disease (DITRA), IL-1 receptor antagonist deficiency autoimmune disease (DIRA), or cryopyrin-associated periodic syndrome (CAPS).

[0124] In some embodiments, the humanized antibody exhibits blocking activity, whereby it reduces IL-33, IL-36, or IL-1 mediated activation by at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% when compared to a comparator antibody control or to the absence of the anti-IL1RAP antibody or antibody fragment of the invention. The ability of the antibody to block the binding of IL-33, IL-36, and IL-1 can be measured using binding assays known in the art. Alternatively, the blocking activity of the antibody can be measured by assaying the biological effects of IL-33, IL-36, and IL-1, such as the production of IL-8, IL-6, or IL-12, to determine whether signal transduction mediated by IL1RAP is inhibited.

[0125] In a further embodiment, the invention provides a humanized anti-IL1RAP antibody having favorable biophysical properties. In one embodiment, the humanized anti-IL1RAP antibody of the invention is present in a buffer solution in at least 90% monomeric form, or at least 92% monomeric form, or at least 95% monomeric form. In a further embodiment, the humanized anti-IL1RAP antibody of the invention remains in a buffer solution in at least 90% monomeric form, or at least 92% monomeric form, or at least 95% monomeric form for one month or four months. In a further embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 170 and the heavy chain sequence of SEQ ID NO: 177 (antibody A1). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 171 and the heavy chain sequence of SEQ ID NO: 178 (antibody A2). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 172 and the heavy chain sequence of SEQ ID NO: 179 (antibody A3). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 173 and the heavy chain sequence of SEQ ID NO: 180 (antibody A4). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 174 and the heavy chain sequence of SEQ ID NO: 181 (antibody A5). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 175 and the heavy chain sequence of SEQ ID NO: 182 (antibody A6). In another embodiment, the humanized antibody of the invention consists of the light chain sequence of SEQ ID NO: 176 and the heavy chain sequence of SEQ ID NO: 183 (antibody A7).

[0126] In some embodiments, the humanized anti-IL1RAP antibody, including antigen-binding fragments thereof, such as heavy and light chain variable regions, comprises the amino acid sequence of residues derived from antibody A1, antibody A2, antibody A3, antibody A4, antibody A5, antibody A6, or antibody A7. In a further embodiment, the invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof that competitively binds to human IL1RAP with an antibody of the invention, such as antibody A1, antibody A2, antibody A3, antibody A4, antibody A5, antibody A6, or antibody A7 described herein. The ability of an antibody or antigen-binding fragment to competitively bind to IL1RAP can be measured using competitive binding assays known in the art. Humanized anti-IL1RAP antibodies may contain specific amino acid substitutions in consensus or germline framework regions. Specific substitutions of amino acid residues at these framework positions may improve various aspects of antibody performance, including binding affinity and / or stability, relative to that seen in humanized antibodies formed by "direct swapping" of CDRs or HVLs into human germline framework regions.

[0127] In some embodiments, the present invention describes monoclonal antibodies having a light chain variable region with the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the present invention describes other monoclonal antibodies having a heavy chain variable region with the amino acid sequence set forth in SEQ ID NO: 2. Placing such CDRs in the FRs of human consensus heavy and light chain variable domains will produce useful humanized antibodies of the present invention. In particular, the present invention provides monoclonal antibodies having the combination of the light chain variable and heavy chain variable regions of SEQ ID NO: 1 and SEQ ID NO: 2, respectively. Such variable regions may be combined with human constant regions.

[0128] In some embodiments, the present invention describes other humanized antibodies having a light chain variable region sequence having an amino acid sequence set forth in any one of SEQ ID NOs: 17 to 66. In some embodiments, the present invention describes other humanized antibodies having a heavy chain variable region sequence having an amino acid sequence set forth in any one of SEQ ID NOs: 67 to 116. In particular, the present invention relates to the nucleic acids represented by SEQ ID NOs: 17 / 67, 18 / 68, 19 / 69, 20 / 70, 21 / 71, 22 / 72, 23 / 73, 24 / 74, 25 / 75, 26 / 76, 27 / 77, 28 / 78, 29 / 79, 30 / 80, 31 / 81, 32 / 82, 33 / 83, 34 / 84, 35 / 85, 36 / 86, 37 / 87, 38 / 88, 39 / 89, 40 / 90, 41 / 91, 42 / 92, 43 / 93, 44 / 99, 45 / 9 The present invention provides monoclonal antibodies having the following combinations of light and heavy chain variable regions: 5, 46 / 96, 47 / 97, ​​48 / 98, 49 / 99, 50 / 100, 51 / 101, 52 / 102, 53 / 103, 54 / 104, 55 / 105, 56 / 106, 57 / 107, 58 / 108, 59 / 109, 60 / 110, 61 / 111, 62 / 112, 63 / 113, 64 / 114, 65 / 115, and 66 / 116. Such variable regions may be combined with human constant regions.

[0129] In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 17 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 17, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 67 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 67. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody. In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 36 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 36, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 86 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 86. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody.

[0130] In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 40 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 40, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 90 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 90. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody.

[0131] In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 47 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 47, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 97 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 97. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody.

[0132] In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 50 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 50, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 100 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 100. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody. In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 51 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 51, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 101 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 101. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody.

[0133] In a further embodiment, the invention relates to an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a humanized light chain variable domain comprising the CDRs of SEQ ID NO: 52 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain light chain amino acid sequence of SEQ ID NO: 52, and a humanized heavy chain variable domain comprising the CDRs of SEQ ID NO: 102 and framework regions having an amino acid sequence at least 90% identical, at least 93% identical, or at least 95% identical to the amino acid sequence of the framework regions of the variable domain heavy chain amino acid sequence of SEQ ID NO: 102. In one embodiment, the anti-IL1RAP antibody is a humanized monoclonal antibody.

[0134] In a further aspect, the invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising a light chain CDR1 (L-CDR1) sequence of any one of SEQ ID NOs: 3 or 6; a light chain CDR2 (L-CDR2) sequence of any one of SEQ ID NOs: 4 or 7; a light chain CDR3 (L-CDR3) sequence of SEQ ID NO: 5; a heavy chain CDR1 (H-CDR1) sequence of any one of SEQ ID NOs: 8, 11, 12, 14; a heavy chain CDR2 (H-CDR2) sequence of any one of SEQ ID NOs: 9, 13, or 15; and a heavy chain CDR3 (H-CDR3) sequence of any one of SEQ ID NOs: 10 or 16. In one aspect, the anti-IL1RAP antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an L-CDR1 as listed above, an L-CDR2 as listed above, and an L-CDR3 as listed above, and a heavy chain variable region comprising an H-CDR1 as listed above, an H-CDR2 as listed above, and an H-CDR3 as listed above.

[0135] In a further aspect, the present invention relates to a light chain CDR1 (L-CDR1) sequence of any one of SEQ ID NOs: 3, 6, 134 (wherein X1=A or T; X2=Q or E; X3=S or N; X4=S or N), or 135 (wherein X1=A or T; X2=Q or E; X3=S or N; X4=S or N); a light chain CDR2 (L-CDR2) sequence of any one of SEQ ID NOs: 4, 7, 140 (wherein X1=S or K, X2=S or T), or 141 (wherein X1=S or K); a light chain CDR3 (L-CDR3) sequence of SEQ ID NO: 5; a light chain CDR4 (L-CDR4) sequence of SEQ ID NOs: 8, 11, 12, 14, 146 (wherein X1=M or I, X2=N or and a heavy chain CDR3 (H-CDR3) sequence of any one of SEQ ID NOs: 10 or 16. The present invention provides an anti-IL1RAP antibody or an antigen-binding fragment thereof comprising a heavy chain CDR1 (H-CDR1) sequence of any one of SEQ ID NOs: 9, 13, or 15, 165 (wherein X1=D or G, X2=A or T, X3=N or A, X4=Q or E, X5=M or K, X6=Q or K, X7=D or G), 166 (wherein X1=D or G; X2=A or T), or 167 (wherein X1=D or G; X2=A or T). In one embodiment, the anti-IL1RAP antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an L-CDR1 as listed above, an L-CDR2 as listed above, and an L-CDR3 as listed above, and a heavy chain variable region comprising an H-CDR1 as listed above, an H-CDR2 as listed above, and an H-CDR3 as listed above.

[0136] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 3, 4, 5, 8, 9, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 3, 4, 5, 11, 9, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 3, 4, 5, 12, 13, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 6, 7, 5, 14, 15, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0137] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 8, 149, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 11, 149, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 12, 161, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 7, 5, 14, 162, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0138] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 142, 153, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 144, 153, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 12, 13, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 7, 5, 14, 15, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0139] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 142, 148, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 144, 148, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 136, 5, 12, 161, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 7, 5, 14, 162, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0140] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 8, 148, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 11, 148, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 12, 161, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 139, 5, 14, 162, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0141] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 8, 149, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 11, 149, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 12, 161, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 139, 5, 14, 162, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0142] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 142, 151, and 10 (KABAT), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 144, 151, and 10 (CCG), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 117, 137, 5, 12, 161, and 10 (CHOTHIA), respectively; or d) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 127, 139, 5, 14, 163, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0143] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer a) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 135, 140, 5, 146, 165, and 10 (KABAT / CCG), respectively; or b) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 135, 140, 5, 12, 166, and 10 (CHOTHIA), respectively; or c) the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 sequences of SEQ ID NOs: 136, 141, 5, 14, 167, and 16 (IMGT), respectively The present invention provides an anti-IL1RAP antibody or antigen-binding fragment thereof comprising:

[0144] In one embodiment, the anti-IL1RAP antibody or antigen-binding fragment thereof comprises a light chain variable region comprising a combination of the L-CDR1, L-CDR2, and L-CDR3 listed above, and a heavy chain variable region comprising a combination of the H-CDR1, H-CDR2, and H-CDR3 listed above. In specific embodiments, it is contemplated that chimeric antibodies having CDR regions switched between these exemplary immunoglobulins (i.e., switching, for example, one or two CDRs from one of the mouse antibodies or humanized antibodies derived therefrom with the analogous CDRs from another mouse antibody or humanized antibody derived therefrom) may produce useful antibodies.

[0145] In certain embodiments, the humanized anti-IL1RAP antibody is an antibody fragment. Various antibody fragments have been broadly discussed above, and there are techniques that have been developed for the production of antibody fragments. Fragments can be derived by proteolytic digestion of intact antibodies (see, e.g., Morimoto et al., 1992, Journal of Biochemical and Biophysical Methods 24:107-117; and Brennan et al., 1985, Science 229:81). Alternatively, fragments can be produced directly in recombinant host cells. For example, Fab'-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab')2 fragments (see, e.g., Carter et al., 1992, Bio / Technology 10:163-167). By another approach, F(ab')2 fragments can be directly isolated from recombinant host cell culture. Other techniques for the production of antibody fragments will be apparent to the skilled practitioner. Thus, in one aspect, the invention provides antibody fragments that comprise one of the combinations of CDRs described herein, in particular L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 described herein. In a further aspect, the invention provides antibody fragments that comprise one of the combinations of variable regions described herein, such as the light chain variable region and heavy chain variable region described herein.

[0146] Certain embodiments include a F(ab')2 fragment of a humanized anti-IL1RAP antibody comprising the light chain sequence of any of SEQ ID NOs: 170, 171, 172, 173, 174, 176, or 176 in combination with the heavy chain sequence of SEQ ID NOs: 177, 178, 179, 180, 181, 182, or 183. Such embodiments may include an intact antibody comprising such a F(ab')2. In some embodiments, the antibody or antibody fragment comprises a constant region that mediates effector functions. The constant region may provide antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC) responses. The effector domain may be, for example, the Fc region of an Ig molecule. The effector domain of the antibody can be from any suitable vertebrate species and isotype. Isotypes from various animal species differ in their ability to mediate effector functions. For example, the ability of human immunoglobulins to mediate CDC and ADCC / ADCP generally follows the order IgM≈IgG1≈IgG3>IgG2>IgG4 and IgG1≈IgG3>IgG2 / IgM / IgG4, respectively. Murine immunoglobulins generally follow the order murine IgM≈IgG3>>IgG, respectively. 2b >IgG 2a >>IgG1 and IgG 2b >IgG 2a In another example, murine IgG 2a mediates ADCC, whereas murine IgG 2a Both IgM and IgM mediate CDC.

[0147] Antibody modification Humanized anti-IL1RAP antibodies and agents may include modifications of the humanized anti-IL1RAP antibody or antigen-binding fragment thereof. For example, it may be desirable to modify the antibody with respect to effector function to enhance the effectiveness of the antibody in treating cancer. One such modification is the introduction of a cysteine ​​residue into the Fc region, thereby allowing interchain disulfide bond formation in this region. Homodimeric antibodies thus generated may have improved internalization capability and / or increased complement-mediated cell killing and / or antibody-dependent cellular cytotoxicity (ADCC). See, e.g., Caron et al., 1992, J. Exp Med. 176:1191-1195; and Shopes, 1992, J. Immunol. 148:2918-2922. Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al., 1993, Cancer Research 53: 2560-2565. Alternatively, an antibody can be engineered to contain dual Fc regions, enhancing complement lysis and ADCC capabilities of the antibody. See Stevenson et al., 1989, Anti-Cancer Drug Design 3: 219-230.

[0148] Antibodies with improved ability to support ADCC have been generated by modifying the glycosylation pattern of their Fc region. H2 This is possible because antibody glycosylation at asparagine residue N297 in the domain is involved in the interaction between IgG and Fcγ receptors, which is a prerequisite for ADCC. Host cell lines have been engineered to express antibodies with altered glycosylation, such as increased branching N-acetylglucosamine or decreased fucose. Reduction of fucose provides a greater enhancement of ADCC activity than does increasing the presence of branching N-acetylglucosamine. Furthermore, enhancement of ADCC by low-fucose antibodies is independent of FcγRIIIa V / F polymorphism.

[0149] Modifying the amino acid sequence of the Fc region of an antibody is an alternative to glycosylation engineering to enhance ADCC. The binding site on human IgG1 for Fcγ receptors has been determined by extensive mutational analysis. This has led to the creation of humanized IgG1 antibodies with Fc mutations that increase binding affinity to FcγRIIIa and enhance ADCC in vitro. Additionally, Fc variants have been obtained with many different permutations of binding properties, for example, improved binding to specific FcγR receptors with unchanged or diminished binding to other FcγR receptors.

[0150] Another embodiment includes an immunoconjugate comprising a humanized antibody or fragment thereof conjugated to a cytotoxic agent such as a chemotherapeutic agent, a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or a fragment thereof), or a radioactive isotope (i.e., a radioconjugate). Chemotherapeutic agents useful in generating such immunoconjugates are described above. Enzymatically active toxins and fragments thereof which can be used to form useful immunoconjugates include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, the trichothecenes, and the like. A variety of radionuclides are available for the production of radioconjugated humanized anti-IL1RAP antibodies. Examples include: 212 Bi, 131 I, 131 In,90 Y, and 186 Includes Re.

[0151] Conjugates of humanized anti-IL1RAP antibodies and cytotoxic or chemotherapeutic agents can be made by known methods using a variety of bifunctional protein coupling agents, such as N-succinimidyl-3-(2-pyridyldithiol)propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipomidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, ricin immunotoxins can be prepared as described in Vitetta et al., 1987, Science 238:1098. Carbon-14 labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotides to antibodies. Conjugates can also be formed using cleavable linkers.

[0152] The humanized anti-IL1RAP antibodies disclosed herein can also be formulated as immunoliposomes. Liposomes containing the antibodies are prepared by methods known in the art, such as those described in Epstein et al., 1985, Proc. Natl. Acad. Sci. USA 82:3688; Hwang et al., 1980, Proc. Natl. Acad. Sci. USA 77:4030; and U.S. Patent Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed, for example, in U.S. Patent No. 5,013,556.

[0153] Particularly useful liposomes can be generated by the reverse phase evaporation method using a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter. Fab' fragments of the antibodies disclosed herein can be conjugated to liposomes by a disulfide exchange reaction as described in Martin et al., 1982, J. Biol. Chem. 257:286-288. Chemotherapeutic agents (such as doxorubicin) can be contained within the liposomes. See, e.g., Gabizon et al., 1989, J. National Cancer Inst. 81(19):1484.

[0154] The antibodies described and disclosed herein may also be used in ADEPT (antibody-directed enzyme prodrug therapy) procedures by conjugating the antibody to a prodrug-activating enzyme that converts a prodrug (e.g., a peptidyl chemotherapeutic agent) into an active anticancer drug. See, for example, WO81 / 01145, WO88 / 07378, and U.S. Patent No. 4,975,278. The enzyme component of the immunoconjugate useful for ADEPT is an enzyme that can act on a prodrug in such a way as to convert it into its more active, cytotoxic form. Specific enzymes that are useful in ADEPT include alkaline phosphatase, for converting phosphate-containing prodrugs to free drugs; arylsulfatases, for converting sulfate-containing prodrugs to free drugs; cytosine deaminase, for converting the non-toxic 5-fluorocytosine to the anticancer drug 5-fluorouracil; proteases such as Serratia protease, thermolysin, subtilisin, carboxypeptidase, and cathepsins (such as cathepsin B and L) for converting peptide-containing prodrugs to free drugs; These include, but are not limited to, D-alanyl carboxypeptidase for converting prodrugs containing substituents, carbohydrate cleaving enzymes such as β-galactosidase and neuraminidase for converting glycosylated prodrugs to free drugs, β-lactamase for converting β-lactam derivatized drugs to free drugs, and penicillin amidases such as penicillin V amidase or penicillin G amidase for converting drugs derivatized at their amine nitrogens with phenoxyacetyl or phenylacetyl groups, respectively, to free drugs. Alternatively, antibodies with enzymatic activity ("abzymes") can be used to convert prodrugs to free active drugs (see, e.g., Massey, 1987, Nature 328: 457-458). Antibody-abzyme conjugates may be prepared by known methods for delivery of abzymes to tumor cell populations, for example by covalently attaching an enzyme to the humanized anti-IL1RAP antibody / heterobifunctional crosslinking reagent discussed above.Alternatively, a fusion protein comprising at least the antigen-binding region of an antibody disclosed herein linked to at least a functionally active portion of an enzyme described above can be constructed using recombinant DNA techniques (see, e.g., Neuberger et al., 1984, Nature 312:604-608).

[0155] In certain embodiments, it may be desirable to use a humanized anti-IL1RAP antibody fragment rather than an intact antibody, for example to increase tissue penetration. It may be desirable to modify the antibody fragment to increase its serum half-life. This can be achieved, for example, by incorporating a salvage receptor binding epitope into the antibody fragment. In one method, the appropriate region of the antibody fragment can be altered (e.g., mutated), or the epitope can be incorporated into a peptide tag, which is then fused to the antibody fragment, either terminally or centrally, for example, by DNA or peptide synthesis. See, for example, WO96 / 32478.

[0156] In other embodiments, covalent modifications of the humanized anti-IL1RAP antibody are also included. Covalent modifications include modifications of cysteinyl, histidyl, lysinyl and amino terminal residues, arginyl, tyrosyl, carboxyl side groups (aspartyl or glutamyl), glutaminyl and asparaginyl residues, or seryl or threonyl residues. Another type of covalent modification involves chemically or enzymatically coupling glycosides to the antibody. Such modifications can be made by chemical synthesis or, where applicable, by enzymatic or chemical cleavage of the antibody. Other types of covalent modifications of the antibody can be introduced into the molecule by reacting targeted amino acid residues of the antibody with organic derivatizing agents capable of reacting with selected side chains or amino- or carboxy-terminal residues.

[0157] Removal of any carbohydrate moieties present on the antibody may be accomplished chemically or enzymatically. Chemical deglycosylation is described by Hakimuddin et al., 1987, Arch. Biochem. Biophys. 259:52 and by Edge et al., 1981, Anal. Biochem., 118:131. Enzymatic cleavage of carbohydrate moieties on the antibody can be achieved by the use of a variety of endo- and exo-glycosidases as described by Thotakura et al., 1987, Meth. Enzymol 138:350.

[0158] Another type of useful covalent modification involves linking the antibody to one of a variety of nonproteinaceous polymers, such as polyethylene glycol, polypropylene glycol, or polyoxyalkylenes, in the manner described in one or more of U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192, and 4,179,337.

[0159] Humanization and Amino Acid Sequence Variants Amino acid sequence variants of the anti-IL1RAP antibody can be prepared by introducing appropriate nucleotide changes into the anti-IL1RAP antibody DNA or by peptide synthesis. Such variants include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequences of the example anti-IL1RAP antibodies herein. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics. Amino acid changes can also alter post-translational processing of the humanized or variant anti-IL1RAP antibody, such as by changing the number or position of glycosylation sites.

[0160] A useful method for identifying certain residues or regions of an anti-IL1RAP antibody that are preferred sites for mutagenesis is called "alanine scanning mutagenesis" as described by Cunningham and Wells (Science, 244:1081-1085 (1989)). Here, a residue or group of target residues is identified (e.g., charged residues such as arg, asp, his, lys, and glu) and replaced with neutral or negatively charged amino acids (typically alanine) to affect the interaction of the amino acid with the IL1RAP antigen. Those amino acid locations that show functional sensitivity to the substitution are then refined by introducing further or other variants at or for the site of substitution. Thus, while the site for introducing amino acid sequence variation is predetermined, the nature of the mutation per se need not be predetermined. For example, to analyze the performance of a mutation at a given site, alanine scanning or random mutagenesis is performed at the target codon or region and the expressed anti-IL1RAP antibody variants are screened for the desired activity.

[0161] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from a single residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues. An example of a terminal insertion comprises an anti-IL1RAP antibody fused to an epitope tag. Other insertional variants of the anti-IL1RAP antibody molecule include the fusion to the N- or C-terminus of the anti-IL1RAP antibody of an enzyme or a polypeptide which increases the serum half-life of the antibody. Another type of variant is an amino acid substitution variant. These variants have at least one amino acid residue in the anti-IL1RAP antibody molecule removed and a different residue inserted in its place. The sites of greatest interest for substitution mutagenesis include the hypervariable regions, although FR alterations are also contemplated. Conservative substitutions are shown in Table 26 under the heading of "preferred substitutions". More substantial changes designated as "exemplary substitutions" or described further below in relation to amino acid classes can be introduced and the products screened if such substitutions result in altered biological activity.

[0162] [Table 31]

[0163] In protein chemistry, it is generally accepted that the biological properties of antibodies can be addressed by selecting substitutions that differ significantly in their effect on (a) maintaining the structure of the polypeptide backbone in the area of ​​the substitution, e.g., sheet or helix conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) maintaining the bulk of the side chains. Naturally occurring residues are divided into groups based on common side chain properties: (1) Hydrophobic: norleucine, met, ala, val, leu, ile; (2) Neutral hydrophilic: cys, ser, thr; (3) Acidic: asp, glu; (4) Basic: asn, gin, his, lys, arg; (5) Residues that affect chain orientation: gly, pro; and (6) Aromatics: trp, tyr, phe. Non-conservative substitutions would involve exchanging a member of one of these classes for another class.

[0164] Any cysteine ​​residues not involved in maintaining the proper conformation of a humanized or variant anti-IL1RAP antibody may also be substituted, generally with serine, to improve the oxidative stability of the molecule, to prevent aberrant cross-linking, or to provide an established point for conjugation to a cytotoxic or cytostatic compound. Conversely, cysteine ​​bond(s) may be added to the antibody to improve its stability (particularly where the antibody is an antibody fragment such as an Fv fragment).

[0165] One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Generally, the resulting variants selected for further development will have improved biological properties relative to the parent antibody from which they were generated. A convenient way to generate such substitutional variants is affinity maturation using phage display. Briefly, several hypervariable region sites (e.g., 6-7 sites) are mutated to generate all possible amino substitutions at each site. The antibody variants thus generated are displayed in a monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage-displayed variants are then screened for their biological activity (e.g., binding affinity). To identify candidate hypervariable region sites for modification, alanine scanning mutagenesis can be performed to identify hypervariable region residues that contribute significantly to antigen binding. Alternatively or additionally, it may be beneficial to analyze a crystal structure of the antigen-antibody complex to identify contact points between the antibody and human IL1RAP. Such contact and neighboring residues are candidates for substitution according to the techniques detailed herein. Once such variants are generated, the panel of variants can be subjected to screening as described herein, and antibodies with superior properties in one or more relevant assays can be selected for further development.

[0166] Another type of amino acid variant of an antibody alters the original glycosylation pattern of the antibody. By "altering" is meant deleting one or more carbohydrate moieties found in the antibody, and / or adding one or more glycosylation sites that are not present in the antibody. In some embodiments, it may be desirable to modify an antibody of the invention to add glycosylation sites. Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used. Thus, to glycosylate a given protein, such as an antibody, the amino acid sequence of the protein is engineered to contain one or more of the tripeptide sequences described above (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).

[0167] Nucleic acid molecules encoding amino acid sequence variants of anti-IL1RAP antibodies are prepared by a variety of methods known in the art, including, but not limited to, isolation from natural sources (in the case of naturally occurring amino acid sequence variants) or preparation by oligonucleotide-mediated (or site-directed) mutagenesis, PCR mutagenesis, and cassette mutagenesis of earlier prepared variants or non-variant forms of anti-IL1RAP antibodies.

[0168] Polynucleotides, Vectors, Host Cells, and Recombinant Methods Other embodiments include isolated polynucleotides comprising sequences encoding a humanized anti-IL1RAP antibody, vectors and host cells containing the polynucleotides, and recombinant techniques for the production of humanized antibodies. The isolated polynucleotides can encode an anti-IL1RAP antibody in any desired form, including, for example, full-length monoclonal antibodies, Fab, Fab', F(ab')2, and Fv fragments, diabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0169] Some embodiments include an isolated polynucleotide comprising a sequence encoding a light chain variable region of an antibody or antibody fragment having the amino acid sequence of any of SEQ ID NOs: 17-66. Some embodiments include an isolated polynucleotide comprising a sequence encoding a heavy chain variable region of an antibody or antibody fragment having the amino acid sequence of any of SEQ ID NOs: 67-116. Some embodiments include an isolated polynucleotide comprising a sequence encoding a light chain variable region of an antibody or antibody fragment having the amino acid sequence of any of SEQ ID NOs: 17, 36, 40, 47, 50, 51, and 52. Some embodiments include an isolated polynucleotide comprising a sequence encoding a heavy chain variable region of an antibody or antibody fragment having the amino acid sequence of SEQ ID NOs: 67, 86, 90, 97, 100, 101, and 102. In one aspect, the isolated polynucleotide sequence encodes an antibody or antibody fragment having light and heavy chain variable regions comprising the amino acid sequences of SEQ ID NO:17 and SEQ ID NO:167, respectively; SEQ ID NO:36 and SEQ ID NO:86, respectively; SEQ ID NO:40 and SEQ ID NO:90, respectively; SEQ ID NO:47 and SEQ ID NO:97, respectively; SEQ ID NO:50 and SEQ ID NO:100, respectively; SEQ ID NO:51 and SEQ ID NO:101, respectively; and SEQ ID NO:52 and SEQ ID NO:102, respectively.

[0170] A polynucleotide comprising a sequence encoding a humanized anti-IL1RAP antibody or a fragment or chain thereof may be fused to one or more regulatory or control sequences, as known in the art, and may be contained in a suitable expression vector or host cell, as known in the art. Each of the polynucleotide molecules encoding the heavy or light chain variable domains may be fused independently to a polynucleotide sequence encoding a constant domain, such as a human constant domain, to allow for the production of an intact antibody. Alternatively, the polynucleotides or portions thereof may be fused together to provide a template for the production of a single chain antibody.

[0171] For recombinant production, a polynucleotide encoding the antibody is inserted into a replicable vector for cloning (amplification of the DNA) or for expression. Many suitable vectors are available for expressing recombinant antibodies. Vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.

[0172] The humanized anti-IL1RAP antibody can also be produced as a fusion polypeptide in which the antibody is fused at the amino terminus of the mature protein or polypeptide to a heterologous polypeptide, such as a signal sequence or other polypeptide with a specific cleavage site. The heterologous signal sequence selected is typically one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For prokaryotic host cells that do not recognize and process the humanized anti-IL1RAP antibody signal sequence, the signal sequence can be substituted by a prokaryotic signal sequence. The signal sequence can be, for example, the alkaline phosphatase, penicillinase, lipoprotein, heat-stable enterotoxin II leader, and the like. For yeast secretion, the native signal sequence may be substituted with a leader sequence derived, for example, from yeast invertase alpha factor (including the Saccharomyces and Kluyveromyces alpha-factor leaders), acid phosphatase, C. albicans glucoamylase, or the signals described in WO 90 / 13646. In mammalian cells, mammalian signal sequences may be used as well as viral secretion leaders, such as the herpes simplex gD signal. DNA for such a precursor region is ligated in reading frame to DNA encoding the humanized anti-IL1RAP antibody.

[0173] Expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors, this sequence enables the vector to replicate independently of the host chromosomal DNA and includes an origin of replication or an autonomously replicating sequence. Such sequences are well known for a variety of bacteria, yeast, and viruses. The origin of replication from the plasmid pBR322 is suitable for most gram-negative bacteria, the 2-ν. plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV, and BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter). Expression and cloning vectors may contain genes encoding selectable markers to facilitate identification of expression. Typical selectable marker genes encode proteins that confer resistance to antibiotics or other toxins, such as ampicillin, neomycin, methotrexate, or tetracycline, or alternatively, complement auxotrophic deficiencies, or in other alternatives, supply specific nutrients not present in complex media, such as a gene encoding D-alanine racemase for Bacilli.

[0174] One example of a selection scheme utilizes a drug that stops the growth of the host cell. Those cells that are successfully transformed with the heterologous gene produce a protein that confers drug resistance and therefore survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid, and hygromycin. Common selectable markers for mammalian cells are DHFR (dihydrofolate reductase), thymidine kinase, metallothionein-I and -II (such as primate metallothionein genes), adenosine deaminase, ornithine decarboxylase, etc., which allow the identification of cells that are capable of taking up the nucleic acid encoding the humanized anti-IL1RAP antibody. Cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium containing methotrexate (Mtx), a competitive antagonist of DHFR. An appropriate host cell when wild-type DHFR is employed is a Chinese hamster ovary (CHO) cell line (e.g., DG44) that is deficient in DHFR activity.

[0175] Alternatively, host cells transformed or co-transformed with DNA sequences encoding an anti-IL1RAP antibody, a wild-type DHFR protein, and another selectable marker such as aminoglycoside 3'-phosphotransferase (APH) (particularly wild-type hosts containing endogenous DHFR) can be selected by growing the cells in medium containing a selection agent for the selectable marker, such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See, e.g., U.S. Patent No. 4,965,199.

[0176] When recombinant production is carried out in yeast cells as host cells, the TRP1 gene present in the yeast plasmid YRp7 (Stinchcomb et al., 1979, Nature 282: 39) can be used as a selectable marker. The TRP1 gene provides a selectable marker for mutant strains of yeast lacking the ability to grow in tryptophan, such as ATCC No. 44076 or PEP4-1 (Jones, 1977, Genetics 85:12). The presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan. Similarly, Leu2p-deficient yeast strains, such as ATCC 20,622 and 38,626, are complemented by known plasmids bearing the LEU2 gene. In addition, vectors derived from the 1.6 μm circular plasmid pKD1 can be used to transform Kluyveromyces yeast. Alternatively, an expression system for large-scale production of recombinant calf chymosin has been reported for K. lactis (Van den Berg, 1990, Bio / Technology 8:135). Stable multicopy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed (Fleer et al., 1991, Bio / Technology 9:968-975).

[0177] Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the nucleic acid molecule encoding the anti-IL1RAP antibody or a polypeptide chain thereof. Promoters suitable for use with prokaryotic hosts include the phoA promoter, β-lactamase and lactose promoter systems, alkaline phosphatase, tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter. Other known bacterial promoters are also suitable. Promoters for use in bacterial systems will also contain a Shine-Dalgarno (SD) sequence operably linked to the DNA encoding the humanized anti-IL1RAP antibody. Many eukaryotic promoter sequences are known. Virtually all eukaryotic genes have an AT-rich region located approximately 25-30 bases upstream from the site where transcription begins. Another sequence found 70-80 bases upstream from the start of transcription of many genes is a CNCAAT region, where N can be any nucleotide. At the 3' end of most eukaryotic genes is an AATAAA sequence that may be a signal for addition of a poly A tail to the 3' end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors.

[0178] Examples of suitable promoting sequences for use with yeast hosts include promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, glucosephosphate isomerase, and glucokinase. Inducible promoters have the added advantage of transcription being controlled by growth conditions. These include yeast promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, derivative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes involved in maltose and galactose utilization. Suitable vectors and promoters for use in yeast expression are further described in EP 73,657. Yeast enhancers are also advantageously used with yeast promoters.

[0179] Humanized anti-IL1RAP antibody transcription from the vector in mammalian host cells is controlled by promoters taken from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retroviruses, hepatitis B virus, and Simian Virus 40 (SV40), from heterologous mammalian promoters, such as the actin promoter or immunoglobulin promoters, or from heat shock promoters, provided that such promoters are compatible with the host cell system. The early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment, which also contains the origin of SV40 viral replication. The immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment. A system for expressing DNA in a mammalian host using bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978. See also Reyes et al., 1982, Nature 297:598-601, which discloses the expression of human p-interferon cDNA in mouse cells under the control of the thymidine kinase promoter from herpes simplex virus. Alternatively, the long terminal repeat of Rous sarcoma virus can be used as a promoter.

[0180] Another useful element that can be used in a recombinant expression vector is an enhancer sequence, which is used to increase the transcription of DNA encoding a humanized anti-IL1RAP antibody by higher eukaryotes. Many enhancer sequences are now known from mammalian genes (e.g., globin, elastase, albumin, α-fetoprotein, and insulin). Typically, however, enhancers from eukaryotic cell viruses are used. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. See also Yaniv, 1982, Nature 297:17-18 for a description of enhancing elements for the activation of eukaryotic promoters. The enhancer can be spliced ​​into the vector at a 5' or 3' position to the humanized anti-IL1RAP antibody coding sequence, but is preferably located at a site 5' from the promoter.

[0181] Expression vectors used in eukaryotic host cells (nucleated cells from yeast, fungi, insects, plants, animals, humans, or other multicellular organisms) may also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5', and sometimes 3', untranslated regions of eukaryotic or viral DNA or cDNA. These regions contain nucleotide segments that are transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the anti-IL1RAP antibody. One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94 / 11026 and the expression vectors disclosed therein. In some embodiments, humanized anti-IL1RAP antibodies can be expressed using the CHEF system. (See, e.g., U.S. Pat. No. 5,888,809; the disclosure of which is incorporated herein by reference.)

[0182] Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above. Suitable prokaryotes for this purpose include eubacteria, such as gram-negative or gram-positive organisms, e.g., Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Enterobacteriaceae, such as B. subtilis and B. licheniformis (e.g., B. licheniformis 41, disclosed in DD 266,710 published April 12, 1989). Examples of suitable E. coli cloning hosts include Bacillus, such as P. cerevisiae, Pseudomonas, such as P. aeruginosa, and Streptomyces. One preferred E. coli cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coli X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.

[0183] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for humanized anti-IL1RAP antibody-encoding vectors. Saccharomyces cerevisiae, the common baker's yeast, is the most commonly used among lower eukaryotic host organisms. However, Schizosaccharomyces pombe; Kluyveromyces hosts such as K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans, and K. marxianus; Yarrowia (EP 402,226); Pichia pastoris; pastors (EP 183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa; Schwanniomyces, such as Schwanniomyces occidentalis; and several other genera, species, and strains, such as filamentous fungi, for example Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts, such as A. nidulans and A. niger, are commonly available and useful herein.

[0184] Suitable host cells for the expression of glycosylated humanized anti-IL1RAP antibodies are derived from multicellular organisms. Examples of invertebrate cells include plant and insect cells, including, for example, numerous baculovirus strains and variants, as well as corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruit fly), and Bombyx mori (silkworm). A variety of virus strains for transfection are publicly available, such as the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used, in particular, for transfection of Spodoptera cells. Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco may also be utilized as hosts.

[0185] In another embodiment, expression of humanized anti-IL1RAP is in vertebrate cells. Propagation of vertebrate cells in culture (tissue culture) has become a routine procedure and techniques are widely available. Examples of useful mammalian host cell lines are the SV40 transformed monkey kidney CV1 line (COS-7, ATCC CRL 1651), human embryonic kidney line (293, or 293 cells subcloned for growth in suspension culture (Graham et al., 1977, J. Gen Virol. 36: 59)), baby hamster kidney cells (BHK, ATCC CCL 10), Chinese hamster ovary cells / -DHFR1 (CHO, Urlaub et al., 1980, Proc. Natl. Acad. Sci. USA 77: 4216; e.g., DG44), mouse Sertoli cells (TM4, Mather, 1980, Biol. Reprod. 23:243-251), monkey kidney cells (CV1 ATCC CCL 70), African green monkey kidney cells (VERO-76, ATCC CRL-1587), human cervical carcinoma cells (HELA, ATCC CCL 2), canine kidney cells (MDCK, ATCC CCL 34), buffalo rat liver cells (BRL 3A, ATCC CRL 1442), human lung cells (W138, ATCC CCL 75), human liver cells (Hep G2, HB 8065), mouse mammary tumor (MMT 060562, ATCC CCL51), TR1 cells (Mather et al., 1982, Annals NY Acad. Sci. 383: 44-68), MRC 5 cells, FS4 cells, and a human hepatocellular carcinoma line (Hep G2).

[0186] Host cells are transformed with the expression or cloning vectors described above for producing a humanized anti-IL1RAP antibody and cultured in conventional nutrient media modified as necessary to induce promoters, select transformants, or amplify the genes encoding the desired sequences.

[0187] The host cells used to produce the humanized anti-IL1RAP antibodies described herein may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma-Aldrich Co., St. Louis, Mo.), Minimum Essential Medium ((MEM), Sigma-Aldrich Co.), RPMI-1640 (Sigma-Aldrich Co.), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma-Aldrich Co.) are suitable for culturing the host cells. In addition, any of the media described in one or more of Ham et al., 1979, Meth. Enz. 58: 44; Barnes et al., 1980, Anal. Biochem. 102: 255; U.S. Pat. No. 4,767,704; U.S. Pat. No. 4,657,866; U.S. Pat. No. 4,927,762; U.S. Pat. No. 4,560,655; U.S. Pat. No. 5,122,469; WO90 / 103430; and WO87 / 00195 may be used as a culture medium for the host cells. Any of these media may be supplemented as necessary with hormones and / or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as gentamicin), trace elements (defined as inorganic compounds typically present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Other supplements may also be included at appropriate concentrations that would be known to one of skill in the art. Culture conditions such as temperature, pH, etc. will be those previously used with the host cell selected for expression and will be apparent to one of skill in the art.

[0188] When using recombinant techniques, antibodies can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the antibody is produced intracellularly, the cells can be disrupted to release the protein as a first step. Particulate debris, whether host cells or lysed fragments, can be removed, for example, by centrifugation or ultrafiltration. Carter et al., 1992, Bio / Technology 10:163-167, describes a procedure for isolating antibodies secreted into the periplasmic space of E. coli. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonyl fluoride (PMSF) for about 30 minutes. Cell debris can be removed by centrifugation. If the antibody is secreted into the medium, the supernatant from such expression systems is generally first concentrated using a commercially available protein concentration filter, such as an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the preceding steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.A variety of methods may be used to isolate antibodies from host cells.

[0189] The antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being a typical purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc domain present in the antibody. Protein A can be used to purify antibodies based on human gamma 1, gamma 2, or gamma 4 heavy chains (see, e.g., Lindmark et al., 1983 J. Immunol. Meth. 62:1-13). Protein G is recommended for all mouse isotypes and for human gamma 3 (see, e.g., Guss et al., 1986 EMBO J. 5:1567-1575). The matrix to which the affinity ligand is attached is most often agarose, although other matrices can be used. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. H3 If the antibody contains a nucleotide sequence similar to that of the nucleotide sequence ...

[0190] After any preliminary purification steps, the mixture containing the antibody of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography, typically performed at low salt concentrations (e.g., from about 0-0.25 M salt) and using an elution buffer at a pH of about 2.5-4.5. Also included are nucleic acids that hybridize under low, medium and high stringency conditions as defined herein to all or a portion (e.g., a portion encoding a variable region) of the nucleotide sequence represented by the isolated polynucleotide sequence encoding an antibody or antibody fragment of the invention. The hybridizing portion of the hybridizing nucleic acid is typically at least 15 (e.g., 20, 25, 30, or 50) nucleotides in length. The hybridizing portion of the hybridizing nucleic acid is at least 80%, e.g., at least 90%, at least 95%, or at least 98% identical to a portion or all of the sequence of a nucleic acid encoding an anti-IL1RAP polypeptide (e.g., a heavy or light chain variable region) or its complement. Hybridizing nucleic acids of the type described herein can be used, for example, as cloning probes, primers, e.g., PCR primers, or diagnostic probes.

[0191] non-therapeutic use The antibodies described herein are useful as affinity purification agents. In this process, the antibodies are immobilized on a solid phase, such as a protein A resin, using methods well known in the art. The immobilized antibodies are contacted with a sample containing the IL1RAP protein (or fragment thereof) to be purified, and the support is then washed with a suitable solvent that will remove substantially all material in the sample except for the IL1RAP protein bound to the immobilized antibody. Finally, the support is washed with another suitable solvent that will release the IL1RAP protein from the antibody.

[0192] Anti-IL1RAP antibodies, e.g., humanized anti-IL1RAP antibodies, are also useful in diagnostic assays to detect and / or quantitate IL1RAP protein, e.g., to detect IL1RAP expression in specific cells, tissues, or serum. Anti-IL1RAP antibodies can be used diagnostically, e.g., to monitor disease onset or progression, e.g., as part of a clinical trial procedure to determine the efficacy of a given treatment and / or prevention regimen. Detection can be facilitated by conjugating the anti-IL1RAP antibody. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, various positron-emitting metals using positron emission tomography, and non-radioactive paramagnetic metal ions. See, e.g., U.S. Pat. No. 4,741,900 for metal ions that can be conjugated to antibodies for use as diagnostics according to the present invention.

[0193] Anti-IL1RAP antibodies can be used in methods for diagnosing an IL1RAP-associated disorder (e.g., a disorder characterized by aberrant expression of IL1RAP) or for determining whether a subject has an increased risk of developing an IL1RAP-associated disorder. Such methods include contacting a biological sample from a subject with an IL1RAP antibody and detecting binding of the antibody to IL1RAP. By "biological sample" is intended any biological sample obtained from an individual, cell line, tissue culture, or other source of cells potentially expressing IL1RAP. Methods for obtaining tissue biopsies and body fluids from mammals are well known in the art.

[0194] In some embodiments, the method may further include comparing the level of IL1RAP in the patient sample with a control sample (e.g., a subject not having an IL1RAP-associated disorder) to determine whether the patient has or is at risk of developing an IL1RAP-associated disorder. In some embodiments, it may be advantageous to label the antibody with a detectable moiety, for example for diagnostic purposes. Numerous detectable labels are available, including radioisotopes, fluorescent labels, enzyme substrate labels, and the like. The label may be indirectly conjugated to the antibody using a variety of known techniques. For example, the antibody may be conjugated with biotin, and any of the three broad categories of labels described above may be conjugated with avidin, or vice versa. Biotin selectively binds to avidin, and thus the label may be conjugated to the antibody in this indirect manner. Alternatively, to achieve indirect conjugation of the label to the antibody, the antibody may be conjugated with a small hapten (such as digoxin), and one of the various types of labels described above is conjugated with an anti-hapten antibody (e.g., an anti-digoxin antibody). Thus, indirect conjugation of the label to the antibody may be achieved.

[0195] Exemplary radioisotope labels include: 35 S, 14 C. 125 I, 3 H, and 131 I. The antibody can be labeled with a radioisotope, for example, using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, 1991, Coligen et al., Ed. Wiley-Interscience, New York, NY, Pubs. Radioactivity can be measured, for example, by scintillation counting. Exemplary fluorescent labels include labels derived from rare earth chelates (europium chelates), or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin, and Texas Red can be used. Fluorescent labels can be conjugated to antibodies by known techniques, such as those disclosed in Current Protocols in Immunology. Fluorescence can be quantified using a fluorometer.

[0196] There are a variety of well-characterized enzyme-substrate labels known in the art (for a review, see, e.g., U.S. Pat. No. 4,275,149). The enzyme generally catalyzes a chemical alteration of a chromogenic substrate that can be measured using a variety of techniques. For example, the alteration can be a color change of the substrate that can be measured spectrophotometrically. Alternatively, the enzyme can alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying the change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and can then emit light or donate energy to a fluorescent acceptor that can be measured, for example, using a chemiluminometer.

[0197] Examples of enzyme labels include luciferases, such as firefly luciferase and bacterial luciferase (U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinedione, malate dehydrogenase, urease, peroxidases, such as horseradish peroxidase (HRPO), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases (such as glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described, for example, in O'Sullivan et al., 1981, Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (J. Langone & H. Van Vunakis, eds.), Academic press, NY, 73: 147-166.

[0198] Examples of enzyme-substrate combinations include, for example, horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, which oxidizes a dye precursor such as orthophenylenediamine (OPD) or 3,3',5,5'-tetramethylbenzidine hydrochloride (TMB); alkaline phosphatase (AP) with para-nitrophenyl phosphate as a chromogenic substrate; and β-D-galactosidase (β-D-Gal) with a chromogenic substrate such as p-nitrophenyl-β-D-galactosidase or the fluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase. Numerous other enzyme-substrate combinations are available to those of skill in the art, for general reviews see U.S. Patent No. 4,275,149 and U.S. Patent No. 4,318,980. In another embodiment, the humanized anti-IL1RAP antibody is used unlabeled and is detected using a labeled antibody that binds to the humanized anti-IL1RAP antibody. The antibodies described herein may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. See, e.g., Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987).

[0199] Anti-IL1RAP antibodies or antigen-binding fragments thereof may be used to inhibit binding of ligands to the IL-36 receptor. Such methods include administering an anti-IL1RAP antibody or antigen-binding fragment thereof to a cell (e.g., a mammalian cell) or cellular environment, whereby signal transduction mediated by the IL-36 receptor is inhibited. These methods may be performed in vitro or in vivo. By "cellular environment" is intended the tissue, medium, or extracellular matrix surrounding the cell. The anti-IL1RAP antibody or antigen-binding fragment thereof is administered to the cellular environment of the cell in such a manner that the antibody or fragment can bind to IL1RAP molecules that are outside and surrounding the cell, thus preventing binding of IL-36 ligand to its receptor.

[0200] Diagnostic Kits The anti-IL1RAP antibodies may be used in diagnostic kits, i.e., packaged combinations of reagents in predetermined amounts with instructions for carrying out a diagnostic assay. When the antibody is labeled with an enzyme, the kit may include substrates and cofactors required by the enzyme, such as substrate precursors that provide a detectable chromophore or fluorophore. In addition, other additives may be included, such as stabilizers, buffers (e.g., blocking buffers or lysis buffers). The relative amounts of the various reagents may vary widely to provide concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. The reagents may be provided as dry powders, typically lyophilized, including excipients that will provide a reagent solution having the appropriate concentration upon dissolution.

[0201] therapeutic use In another embodiment, the humanized anti-IL1RAP antibodies disclosed herein are useful in treating various disorders associated with expression of IL1RAP as described herein. A method for treating an IL1RAP-associated disorder comprises administering to a subject in need thereof a therapeutically effective amount of a humanized anti-IL1RAP antibody. The humanized anti-IL1RAP antibody or agent is administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, as well as intralesional administration (including perfusing or otherwise contacting the graft with the antibody prior to transplantation) if desired for localized immunosuppressive treatment. The humanized anti-IL1RAP antibody or agent may be administered, for example, as an infusion or as a bolus. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, the humanized anti-IL1RAP antibody is suitably administered by pulse infusion, particularly with descending doses of antibody. In one embodiment, dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether administration is brief or chronic.

[0202] With regard to preventing or treating disease, the appropriate dosage of the antibody will depend on a variety of factors, such as the type of disease being treated as defined above, the severity and course of the disease, whether the antibody is being administered for preventive or therapeutic purposes, previous therapy, the patient's medical history and response to the antibody, and the discretion of the attending physician. The antibody is suitably administered to the patient at one time or over a series of treatments. Depending on the type and severity of the disease, about 1 μg / kg to 20 mg / kg (e.g., 0.1 to 15 mg / kg) of antibody is an initial candidate dosage for administration to a patient, whether by one or more separate administrations or by continuous infusion, for example. Typical daily dosages may range from about 1 μg / kg to 100 mg / kg or more, depending on the factors mentioned above. For repeated administration over several days or longer, depending on the condition, treatment is continued until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays. Exemplary dosage regimens are those disclosed in WO94 / 04188. The term "suppression" is used herein in the same context as "amelioration" and "alleviation" to mean a lessening of one or more characteristics of a disease.

[0203] The antibody compositions will be formulated, dosed, and administered in a manner consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the schedule of administration, and other factors known to medical practitioners. A "therapeutically effective amount" of the antibody to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disorder associated with IL1RAP expression. The antibodies need not be, but may be, formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents will depend on the amount of humanized anti-IL1RAP antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. These will generally be used in the same dosages and using the routes of administration used above, or at about 1-99% of the dosages previously employed.

[0204] Pharmaceutical Compositions and Their Administration A composition comprising an IL1RAP binding agent (e.g., an anti-IL1RAP antibody) may be administered to a subject having or at risk of having an immunological disorder, a respiratory disorder, or cancer. The present invention further provides for the use of an IL1RAP binding agent (e.g., an anti-IL1RAP antibody) in the manufacture of a medicament for preventing or treating cancer, a respiratory disorder, or an immunological disorder. As used herein, the term "subject" refers to any mammalian patient to which an IL1RAP binding agent may be administered, including, for example, humans and non-human mammals, such as primates, rodents, and dogs. Subjects specifically intended for treatment using the methods described herein include humans. The antibody or agent may be administered alone or in combination with other compositions in preventing or treating an immunological disorder, a respiratory disorder, or cancer. Such compositions that may be administered in combination with an antibody or agent include methotrexate (MTX), and immunomodulatory agents, such as antibodies or small molecules.

[0205] Examples of antibodies for use in such pharmaceutical compositions include antibodies or antibody fragments having a light chain variable region amino acid sequence of any of SEQ ID NOs: 17 to 66. Examples of antibodies for use in such pharmaceutical compositions also include humanized antibodies or antibody fragments having a heavy chain variable region amino acid sequence of any of SEQ ID NOs: 67 to 116. Further examples of antibodies for use in such pharmaceutical compositions also include humanized antibodies or antibody fragments having the light chain variable region amino acid sequence of any of SEQ ID NOs: 17, 36, 40, 47, 50, 51, and 52. Preferred antibodies for use in such pharmaceutical compositions also include humanized antibodies or antibody fragments having the heavy chain variable region amino acid sequence of any of SEQ ID NOs: 67, 86, 90, 97, 100, 101, and 102. Further examples of antibodies for use in such pharmaceutical compositions also include humanized antibodies or antibody fragments having the light chain variable region and heavy chain variable region of any of SEQ ID NOs: 17 and 67, SEQ ID NOs: 36 and 86, SEQ ID NOs: 40 and 90, SEQ ID NOs: 47 and 97, SEQ ID NOs: 50 and 100, SEQ ID NOs: 51 and 101, and SEQ ID NOs: 52 and 102. Further examples of antibodies for use in such pharmaceutical compositions also include antibody A1, antibody A2, antibody A3, antibody A4, antibody A5, antibody A6, or antibody A7.

[0206] A variety of delivery systems are known and can be used to administer the IL1RAP binding agent. Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The IL1RAP binding agent can be administered, for example, by infusion, bolus, or injection, and can be administered together with other biologically active agents, such as chemotherapeutic agents. Administration can be systemic or local. In a preferred embodiment, administration is by subcutaneous injection. Formulations for such injections can be prepared, for example, in prefilled syringes, which can be administered once every other week. In specific embodiments, the IL1RAP binding agent composition is administered by injection, by catheter, by suppository, or by implant, where the implant is of a porous, non-porous, or gelatinous material, including a membrane, such as a sialastic membrane, or a fiber. Typically, when administering the composition, a material is used that does not absorb the anti-IL1RAP antibody or agent.

[0207] In other embodiments, the anti-IL1RAP antibody or agent is delivered in a controlled release system. In one embodiment, a pump can be used (see, e.g., Langer, 1990, Science 249:1527-1533; Sefton, 1989, CRC Crit. Ref. Biomed. Eng. 14:201; Buchwald et al., 1980, Surgery 88:507; Saudek et al., 1989, N. Engl. J. Med. 321:574). In another embodiment, a polymeric material is used. (See, e.g., Medical Applications of Controlled Release (Langer and Wise eds., CRC Press, Boca Raton, Fla., 1974); Controlled Drug Bioavailability, Drug Product Design and Performance (Smolen and Ball eds., Wiley, New York, 1984); Ranger and Peppas, 1983, Macromol. Sci. Rev. Macromol. Chem. 23:61. See also Levy et al., 1985, Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 71:105.) Other controlled release systems are discussed, e.g., in Langer, supra. An IL1RAP binding agent (eg, an anti-IL1RAP antibody) can be administered as a pharmaceutical composition comprising a therapeutically effective amount of the binding agent and one or more pharma- ceutically compatible ingredients.

[0208] In a typical embodiment, the pharmaceutical composition is formulated according to routine procedures as a pharmaceutical composition suitable for intravenous or subcutaneous administration to humans. Typically, compositions for administration by injection are solutions in sterile isotonic aqueous buffer. If necessary, the medicament may also include a solubilizing agent and a local anesthetic, such as lignocaine, to ease pain at the site of the injection. Generally, the ingredients are supplied separately or mixed together in unit dosage form, for example as a dry lyophilized powder or water-free concentrate, in a sealed container, such as an ampoule or sachette, indicating the quantity of active agent. If the medicament is to be administered by injection, it can be dispensed using an infusion bottle containing sterile pharmaceutical grade water or saline. If the medicament is to be administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

[0209] Additionally, the pharmaceutical composition may be provided as a pharmaceutical kit comprising (a) a container containing an IL1RAP binding agent (e.g., an anti-IL1RAP antibody) in lyophilized form, and (b) a second container containing a pharma- ceutically acceptable diluent for injection (e.g., sterile water). The pharma- ceutically acceptable diluent may be used to reconstitute or dilute the lyophilized anti-IL1RAP antibody or agent. Such containers may optionally be accompanied by a notice in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceutical or biological products, the notice reflecting approval by the agency for manufacture, use, or sale for human administration. The amount of an IL1RAP binding agent (e.g., an anti-IL1RAP antibody) that is effective in treating or preventing an immunological disorder or cancer can be determined by standard clinical techniques. In addition, in vitro assays can be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the stage of the immunological disorder or cancer, and should be decided according to the judgment of the practitioner and each patient's circumstances. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0210] In general, the dosage of an anti-IL1RAP antibody or IL1RAP binding agent administered to a patient with an immunological disorder or an IL1RAP-expressing cancer is typically about 0.1 mg / kg to about 100 mg / kg of the subject's body weight. The dosage administered to a subject is about 0.1 mg / kg to about 50 mg / kg, about 1 mg / kg to about 30 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 15 mg / kg, or about 1 mg / kg to about 10 mg / kg of the subject's body weight.

[0211] Exemplary doses include, but are not limited to, 1 ng / kg to 100 mg / kg. In some embodiments, the dose is about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, or about 16 mg / kg. The dose can be administered, for example, daily, once per week (every week), twice per week, three times per week, four times per week, five times per week, six times per week, once every two weeks, or monthly, bimonthly, or trimonthly. In specific embodiments, the dose is about 0.5 mg / kg / week, about 1 mg / kg / week, about 2 mg / kg / week, about 3 mg / kg / week, about 4 mg / kg / week, about 5 mg / kg / week, about 6 mg / kg / week, about 7 mg / kg / week, about 8 mg / kg / week, about 9 mg / kg / week, about 10 mg / kg / week, about 11 mg / kg / week, about 12 mg / kg / week, about 13 mg / kg / week, about 14 mg / kg / week, about 15 mg / kg / week, or about 16 mg / kg / week. In some embodiments, the dose ranges from about 1 mg / kg / week to about 15 mg / kg / week.

[0212] In some embodiments, a pharmaceutical composition comprising an IL1RAP binding agent may further comprise a therapeutic agent conjugated or unconjugated to the binding agent. The anti-IL1RAP antibody or IL1RAP binding agent may be co-administered in combination with one or more therapeutic agents for the treatment or prevention of an immunological disorder or cancer. Such combined therapeutic administration may have additive or synergistic effects on disease parameters (eg, severity of symptoms, number of symptoms, or frequency of recurrence). With respect to therapeutic regimens for combined administration, in a specific embodiment, the anti-IL1RAP antibody or IL1RAP binding agent is administered simultaneously with the Therapeutic Agent. In another specific embodiment, the Therapeutic Agent is administered at least one hour and up to several months before or after administration of the anti-IL1RAP antibody or IL1RAP binding agent, e.g., at least one hour, five hours, twelve hours, one day, one week, one month, or three months before or after administration of the anti-IL1RAP antibody or IL1RAP binding agent.

[0213] manufactured goods In another embodiment, an article of manufacture is included that contains materials useful for treating the disorders described above. The article of manufacture includes a container and a label. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials, such as glass or plastic. The container holds a composition that is effective for treating a medical condition and can have a sterile access port. For example, the container can be an intravenous fluid bag or a vial with a stopper that can be pierced by a hypodermic needle. The active agent in the composition is a humanized anti-IL1RAP antibody. A label on or associated with the container indicates that the composition is used to treat a selected medical condition. The article of manufacture can further include a second container that contains a pharma- ceutically acceptable buffer, such as phosphate buffered saline, Ringer's solution, and dextrose solution. It can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

[0214] The present invention is further described in the following examples, which are not intended to limit the scope of the invention. The antibodies of the invention are further described in the Examples below. EXAMPLES

[0215] Example 1 Identification of anti-human IL1RAP antibodies A) Immunization with human IL1RAP Mice were immunized with recombinantly produced human IL1RAP (Genbank accession NP_002173.1) protein and produced strong titer responses that were incorporated into traditional hybridoma production. Fusion products that block IL33 and IL36 signaling in whole blood and PBMCS assays with weaker / cell type specific activity against IL1 were subcloned and rescreened. Inhibition of each of the cytokines that signal through IL1RAP is currently being investigated in clinical studies using either antibodies against individual cytokines or against the cognate receptors, demonstrating the appeal of this pathway family. Variable domains were cloned from hybridomas using standard PCR primer sets. The variable domains and specific CDRs are described above.

[0216] B) Generation of chimeric Fab: Candidate IL1RAP G011 chimeric Fabs were generated to benchmark and evaluate humanized / sequence-optimized Fabs in ELISA binding experiments. Briefly, chimeric Fabs were generated by fusing mouse VK and VH residues to human Ck and CH1 residues, respectively. Using the mouse sequences of selected antibody candidates, gene fragments were designed and synthesized for murine VK and VH. The V regions were amplified using a biotinylated forward primer containing a specific sequence for the murine framework 1 region and an overhang sequence annealed to the end of the gene III sequence, and a reverse primer from the conserved human constant region (Ck or CH1, respectively). PCR was performed using the gene fragments as templates, and the DNA products were cloned into M13LE01 vectors using standard protocols. E. coli plaques expressing the DNA of interest were selected and the corresponding DNA samples were isolated and sequenced. Exponentially grown E. coli XL1-blue were induced with 0.5 mM IPTG followed by infection with the correct plaque representing the chimeric Fab and grown overnight at 25°C. Either culture supernatant or bacterial pellet was collected by centrifugation. The pellet was frozen at -80°C for 15 min followed by thawing at RT in lysis buffer (20 mM Tris, 150 mM NaCl, 5 mM EDTA, pH 8.0). The pellet was resuspended by vigorous vortexing for 15 min. Lysates from periplasmic extracts containing soluble chimeric Fab were obtained by centrifugation.

[0217] C) ELISA Binding Assay Development: To identify positive binders during screening, an ELISA binding assay was developed to identify binders independent of Fab expression and ensure that the data represented the binding activity of the Fab. Briefly, plates were coated overnight with various amounts of anti-Fd (Meridian Life Science, Cat#W90075C) in a 96-well format. Assay plates were blocked with 3% milk in PBD for 1 hour, followed by the addition of various amounts of either culture supernatant or periplasmic extract of chimeric / sequence-optimized Fab. For primary screening, plates were coated with 800ng / mL of anti-Fd antibody and for confirmation screening with 400ng / mL of anti-Fd antibody (periplasmic Fab, triplicate). Biotinylated antigen was used at 30ng / mL for 1× primary screening (secreted Fab) and 10ng / mL for 3× confirmation screening (periplasmic Fab, triplicate).

[0218] D) Cloning, expression, and purification of IgG The mouse antibody was converted to a chimeric antibody consisting of a mouse variable domain on a human constant domain (hu IgG1KO / kappa). The hu IgG1KO (knock out) has two replacement mutations (Leu234Ala and Leu235Ala) that eliminate ADCC and CDC activity by reducing effector functions such as FcγR and complement binding. The variable domains of the mouse and chimeric antibodies are identical. The chimeric antibody was generated to verify the function of the antibody and ensure that the correct variable domain sequences were obtained. To generate the antibody constructs, the codon-optimized DNA for the sequence-optimized VH and VK regions was fused to the heavy and light chain constant regions from human IgG1 and directionally cloned into the pTT5 huIgK vector and the VH gene into the pTT5 huIgG1KO vector using the In-Fusion® HD Cloning Kit (Clontech, USA). To facilitate In-Fusion® HD Cloning, the PCR products were purified and treated with Cloning Enhancer before In-Fusion® HD Cloning. Cloning and transformation were performed according to the manufacturer's protocol (Clontech, USA). The DNA sequences of the subcloned gene V gene fragments were confirmed by isolating miniprep DNA and subjecting the isolated DNA to Sanger double-stranded sequencing.

[0219] Both light and heavy chain expression vector DNAs were expanded for transfection into CHO-E cells. Transfected CHO-E cells growing in suspension in serum-free medium were cultured in shake flasks under agitation at 140 rpm, 37°C, and 5% CO2 and kept in exponential growth conditions. On the day of transfection, cells were chemically transfected with 1 mg of light chain plasmid and 0.5 mg of heavy chain plasmid. They were then cultured at 1-2 x 10 cells / ml in 1 L of Gibco® FreeStyle™ CHO Expression Medium (LifeTechnologies, NY, US). 6 Cells were seeded at 1000 cells / ml. Cells were then incubated under orbital shaking with a single supply of 150 ml of commercial feed solution for 10-12 days to allow protein expression. Antibody titers in cell culture supernatants were determined using an Octet® instrument (Pall ForteBio, CA, US) and a protA biosensor chip according to the manufacturer's instructions.

[0220] Recombinant antibodies were purified from culture supernatants by Protein A affinity chromatography using MabSelect™ (Amersham Biosciences) and stored in 60 mM NaOAc buffer (pH 5.0). The purity and degree of heterogeneity of the samples was assessed by mass spectrometry and analytical ultracentrifugation. Prior to functional testing, all samples were confirmed to have a monomer content of ≧90% and contain <10% impurities.

[0221] Example 2 Production of humanized IL1RAP antibodies To reduce potential immunogenicity following administration in humans, the murine anti-human IL1RAP monoclonal antibody GO11 was "humanized" by a design and screening process. Human framework sequences were selected for the murine lead based on framework homology, CDR structure, conserved canonical residues, conserved interface packing residues, and other parameters. Specific substitution of amino acid residues at these framework positions may improve various aspects of antibody performance, including binding affinity and / or stability, compared to those exhibited in humanized antibodies formed by "direct swapping" of CDRs or HVLs into human germline framework regions. For framework engineering, we identified closely matching human germline for the murine GO11 candidate and grafted the CDRs from the murine lead onto the human germline IGKV1-39*01 and IGHV1-69*02. The resulting grafted variable regions were cloned into M13-based vectors and expressed in E. coli cells. The grafted Fabs from E. coli periplasmic extracts were assessed for binding activity in ELISA experiments, and a chimeric Fab with mouse variable regions was kept as a positive control (Figure 1). As can be seen in FIG. 1, the grafted Fab portrayed weaker binding activity than the mouse Fab, suggesting the need to identify critical mouse residues to regain binding activity.

[0222] Besides the CDRs, certain mouse framework residues, such as canonical and Vernier zone residues, are known to play important roles in positioning the CDR loops. To identify human frameworks that retain similar binding of murine lead candidate residues closer to the CDRs, the Vernier zone and canonical residues were changed from human to mouse residues. Phage libraries representing various mutations / changes in the framework sequences were constructed, specifically K45Q, I48V, Y49H, and Q100A in VK, while Q3H, K23E, R38K, A40R, M48I, R67K, V68A, T69R, I70F, A72V, and Y95F for VH were evaluated (Figure 2A / D). E. coli cells were transformed with DNA containing the various combinations. Approximately 450 VK clone variants and 1350 VH clone variants were selected and screened for binding to huIL1RAP by ELISA in comparison to the chimeric parent anti-IL1RAP Fab.

[0223] Molecules that depicted similar binding to the positive control were selected and checked again for their binding activity in a confirmatory ELISA binding experiment. Plaques representing the selected binders were used to reinfect E. coli cells and the cultures were induced with 0.5 mM IPTG. Periplasmic extracts were used to confirm binding to the huIL1RAP antigen. As before, we kept the chimeric Fab as a positive control in our experiments.

[0224] As can be seen in Figures 2A-D, clones 405-10 and 405-12 (SEQ ID NOs: 18 and 19, respectively) for VK and clones 406-18 and 406-20 (SEQ ID NOs: 68 and 69, respectively) for VH depicted binding similar to the chimeric control. Sequence analysis of these light chain frameworks revealed that the amino acid mutation Y49H in FW2 was present in all clones that depicted binding similar to the positive control chimeric Fab and is therefore considered an important critical murine residue to be included in the optimized light chain framework. For VH, the amino acid mutation A72V in FW3 was present in multiple clones, including 406-18 and 406-20, along with other murine residues, and therefore considered important, but not critical, for the optimized molecule. 406-18 had an additional change T69R, while 4306-20 had an I70F change in framework 3 in addition to A72V, but neither of these residues appear to be critical for binding. Given these observations, we decided to use the grafted VH with the A72V residue as a potential critical residue in the final library and as described later.

[0225] CDR Optimization Germlining has evolved as a novel strategy to change non-critical mouse residues to more human germline residues to improve the percent human content of antibodies derived from non-human primates and thus reduce the formation of anti-drug antibodies. We evaluated the role of these mouse residues of IL1RAP in L-CDR1, L-CDR2, and part of L-CDR3, H-CDR1, and H-CDR2 by changing them to human germline residues as point mutations in the IgG backbone. Thirteen variants for the light chain (T25A, E27Q, N28S, N30S, Y50A, K52S, T53Y, A55Q, E56S, H90Q, H91S, G93S, and S95P) and 19 variants for the heavy chain (Y27G, I28T, L30S, T31S, W33Y, M34I, N35S, Q50R, F52I, A54I, S55L, D56G, S57I, T58A, N61A, E62Q, M63K, K65Q, and D66G) were generated separately in the vector pTT5 (see Figure 3A / B). For expression analysis in CHO-E cells for 7 days, the light chain variants were paired with the parent chimeric heavy chain and the heavy chain variants were paired with the parent chimeric light chain. The resulting variants were evaluated for binding in ELISA experiments and compared to the parent chimeric IgG with mouse variable regions but human constant regions. For the light chain variants, the six point variants VK-T25A, E27Q, N28S, N30S, K52S, and T53Y depicted binding similar to the chimeric IgG (+10% control; Figure 3A), while for the heavy chain, the ten point variants VH-I28T, L30S, M34I, N35S, D56G, T58A, N61A, E62Q, M63K, K65Q, and D66G depicted binding similar to the chimeric IgG (+10% control; Figure 3B). Fabs that showed better or equal binding and improved expression compared to the chimeric parent Fab were selected for further characterization.

[0226] Representative humanized variable regions for antibody GO11 are shown in Section 2 of the specification. Thus, antibodies A1 to A7 were humanized antibodies derived from murine antibody 005-GO11 (VL / VH SEQ ID NOs: 1 and 2) (cloned into a human IgG1 KO (KO = knockout) / kappa backbone). Antibodies A1 to A7 are shown in Table 25. Exemplary anti-IL1RAP Abs of the invention specifically bind human and cyno IL1RAP (ECD). The binding affinity of the antibodies binding to human IL1RAP and cyno IL1RAP was determined to be 110 and 160 pM, respectively, using SPR. The anti-IL1RAP antibodies of the invention are not cross-reactive with mouse IL1RAP.

[0227] [Table 32]

[0228] Example 3 Biological activity; Potency of humanized anti-human IL1RAP antibodies in functional human assays - Blockade of IL-33, IL-36, and IL-1 signaling Activation of target cells by IL-1β depends on its interaction with two membrane-bound receptors: IL-1 receptor type I (IL-1R1) and IL-1 receptor accessory protein (IL-1RAcP). IL-1R1 is a ligand-recognition receptor that binds IL-1β with high affinity. Although IL-1RAcP does not directly interact with IL-1β, its recruitment is essential for the formation of a signaling-competent complex. IL1RAP is also a functional part of the IL-33 receptor complex, and therefore, binding to IL1RAP can inhibit both IL-1 and IL-33 signaling. To test the ability of potential lead candidate antibodies to block IL-1 and IL-33 signaling, IL-1 and IL-33 dependent cell assays were established. Exemplary antibodies of the invention were tested for inhibition of IL-1β and IL-33 signaling.

[0229] Protocol: Generation and stimulation of monocyte-derived macrophages (MDMs): Human monocyte-derived macrophages were differentiated and generated as recently described by Koss et al. (IL36 is a Critical Upstream Amplifier of Neutrophilic Lung Inflammation in Mice, publication pending). Cells were stimulated with medium containing Dulbecco's Modified Eagle Medium (DMEM) (1×) + GlutaMAX™-I (GIBCO #31966-021); 10% HI fetal calf serum (FCS; GIBCO #16140-071); 1% NEAA (100× GIBCO #11140-035); 1% P / S (10,000 U / mL penicillin, 10,000 μg / mL streptomycin, GIBCO #15140-122), and 10 ng / mL recombinant human MCSF (R&D Systems; 216-MCC / CF). 6.2×104 MDMs were seeded in 96-well plates. After 24 h, macrophages were pre-stimulated with anti-IL1rap AB (0.4, 2, 10, 50, 250 nM), and 30 min later, MDMs were stimulated with rhIL-36α (33 ng / mL; 6995-IL-010 / CF; R&D Systems), rhIL-36β (33 ng / mL; 6834-ILB-025 / CF; R&D Systems), rhIL-36γ (33 ng / mL; 2320-IL-025 / CF; R&D Systems), or rhIL-1α (10 ng / mL; 200-LA-010 / CF; R&D Systems) for 24 h at 37° C. and 5% CO2. IL12p40 concentrations were measured in the supernatants by a single MSD (Meso Scale Discovery, #K151UQK-1) according to the manufacturer's instructions.

[0230] Generation and stimulation of monocyte-derived dendritic cells (MoDCs): Monocytes were isolated from peripheral blood mononuclear cells from human volunteer donors as described in Koss et al. (IL36 is a Critical Upstream Amplifier of Neutrophilic Lung Inflammation in Mice, publication pending). 5Monocytes were seeded in 96-well plates using a Dendritic Cell Culture Kit (#10985, Stemcel) according to the manufacturer's instructions. After 8 days, differentiated MoDCs were pre-stimulated with anti-IL1rap AB (0.4, 2, 10, 50, 250 nM) for 30 min and further stimulated with rhIL-36α (33 ng / mL; 6995-IL-010 / CF; R&D Systems), rhIL-36β (33 ng / mL; 6834-ILB-025 / CF; R&D Systems), and rhIL-36γ (33 ng / mL; 2320-IL-025 / CF; R&D Systems) for 48 h at 37°C and 5% CO2. IL-8, IL-6, and TNFα concentrations were measured in the supernatants by MSD (Meso Scale Discovery, #K15067L) according to the manufacturer's instructions.

[0231] Whole Blood Assays: 180 μL of whole blood from human volunteer donors (diluted 1:2 with RPMI medium (RPMI1640 Gibco #61870-010)) was pipetted into a 96-well plate. Anti-IL1rap Abs of the present invention (0.07, 0.21, 0.62, 1.8, 5.5, 16.5, 50, 150, 450 nM (MIP1β), 0.1, 1, 10, 100, 1000 nM (IFNγ)) were added to the wells and pre-incubated for 30 minutes at 37° C. and 5% CO2. rhIL33 (3625-IL-010 / CF, R&D Systems, 0.6 nM), rhIL36γ (6835-IL-010 / CF, R&D Systems, 10 nM), and rhIL-12 (219-IL / CF, R&D Systems, 0.25 nM) were added to the wells for 24 h at 37°C and 5% CO2. Supernatants were collected and Mip1β (DY271-05, DuoSet ELISA, R&D Systems) and IFNγ (#555142, ELISA, BD Biosciences) protein concentrations were measured according to the manufacturer's instructions.

[0232] result: The ability of anti-IL1RaP Ab to suppress immune cell activation by IL1 family cytokines was determined. Human monocyte-derived macrophages (MDMs) were stimulated with IL-36α, β, γ alone, IL-1a alone, or a combination of IL-36α, β, γ, and IL-1a as relevant human innate immune cells in vitro. IL-12 protein production was measured as a marker of cell activation downstream of IL1 family cytokine activation. The production of IL-12 in response to IL-36α, β, γ, or IL-36α, β, γ in combination with IL-1a was significantly inhibited by anti-IL1RaP Ab (IC50=0.35nM and IC50=6.48nM, respectively) (FIG. 4A). FIG. 4A shows the inhibition of IL-12p40 secretion by the anti-IL1RaP Ab of the present invention in cytokine-stimulated MDM (mean values ​​± SD of technical triplicates from one representative of two experiments with MDM obtained from different donors are depicted). Since IL-1a alone could not induce IL-12 production in MDM and similar results were obtained after IL-33 stimulation (data not shown), the ability of anti-IL1RaP Ab to suppress IL-12 protein was measured in response to stimulation with IL-4 followed by exposure to IL-36α, β, γ, IL-1a, or IL-36α, β, γ with IL-1a. Anti-IL1RaP Ab #A2 significantly suppressed IL-12 production in response to IL-36α, β, γ (IC50 = 1.91 nM), IL-1a (IC50 = 5.92 nM), and IL-36α, β, γ / IL-1a (IC50 = 8.14 nM) in IL-4-conditioned MDMs (Figure 4A).

[0233] Previous experiments demonstrated that MoDCs were unresponsive to stimulation with IL-1a or IL-33 (not shown). Therefore, as a next measure of the potency of the IL1RAP Ab, we stimulated human monocyte-derived dendritic cells (MoDCs), relevant human adaptive immune cells in vitro, with IL-36α, β, γ, or alone. Protein production of IL-8 (IC50=3.95 nM), TNF (IC50=6.45 nM), and IL-6 (IC50=9.41 nM) was significantly suppressed by anti-IL1Rap Ab (Figure 4B). Consistent with the findings in MDM and DC, when we measured the production of the standard myeloid cell-derived mediator Mip-1b in whole blood, IL-33 did not induce its production, whereas IL-36γ potently induced Mip-1b production, which was significantly inhibited by anti-IL1RaP Ab #A2 (IC50=4.37) (Fig. 4C). Similar data were obtained after combining IL-36γ with IL-33 (IC50=3.43) (Fig. 4C). IL-33 has been reported to activate the production of IFNγ by lymphocytes. Pilot experiments showed that the highest production of IFNγ in whole blood assay was observed after combining IL-33 with IL-12 (data not shown). Therefore, we determined the ability of anti-IL1RaP Ab to inhibit IL-33 / IL-12-mediated production of IFNγ in whole blood assay, and the production of IFNγ was significantly inhibited (IC50=4.84nM) (Figure 4D).

[0234] Example 4 Epitope binding of anti-IL1RAP antibodies: Unique epitopes revealed by crystal structure and HDX MS To understand where anti-IL1RAP clones bind to IL1RAP, structural analysis of the protein was performed using the extracellular domain of the receptor. The "extracellular domain" of IL1RAP forms a cell membrane-bound complex with IL1 receptors, such as IL1R type I or type II. Human IL1RAP (Accession No. Q9NPH3 UniProtKB / Swiss-Prot) is known to consist of three extracellular domains: domain I: amino acids 21-134; domain II: amino acids 135-234; and domain III: amino acids 235-367.

[0235] Crystallization of IL1RAP:Fab binary complex: IL1RAP and anti-IL1RAP #A2 Fab were mixed at a molar ratio of 1.2:1 and concentrated to 20 mg / mL. Crystals were obtained using the hanging drop vapor diffusion method by mixing 1 μl of the complex with 1 μL of reservoir solution containing 0.1 M ammonium acetate, 0.1 M BIS-TRIS pH 6.0, and 15% w / v polyethylene glycol 10000. Plate-like shaped crystals appeared within a day. Crystals were cryo-protected in reservoir solution supplemented with 30% v / v glycerol and flash frozen in liquid nitrogen.

[0236] Data collection and structure determination: Data were collected at beamline X10SA at SLS, Villigen, Switzerland, at 100 K. 720 frames with 0.25° per frame were collected on a PILATUS 6M-F detector. Data were processed with autoPROC software (Smart, et al. Global phasing Limited, 2013;Evans, P. (2006) Scaling and assessment of data quality. Acta Crystallogr D Biol Crystallogr 62, 72-82;Kabsch, W. (2010) Xds. Acta Crystallogr D Biol Crystallogr 66, 125-132;Vonrhein, C., Flensburg, C., Keller, P., Sharff, A., Smart, O., Paciorek, W., Womack, T., and Bricogne, G. (2011) Data processing and analysis with the autoPROC toolbox. Acta Crystallogr D Biol Crystallogr 67, 293-302;Tickle, I., Flensburg, C., Keller, P., Paciorek, W., Sharff, A., Vonrhein, C., and Bricogne, G. (2018) STARANISO. Global Phasing Ltd., Cambridge, United Kingdo;Winn, MD, Ballard, CC, Cowtan, KD, Dodson, EJ, Emsley, P., Evans, PR, Keegan, RM, Krissinel, EB, Leslie, AG, McCoy, A., McNicholas, SJ, Murshudov, GN, Pannu, NS, Potterton, EA, Powell, HR, Read, RJ, Vagin, A., and Wilson, KS.(2011) Overview of the CCP4 suite and current developments. Acta Crystallogr D Biol Crystallogr 67, 235-24; and Evans, PR, and Murshudov, GN (2013) How good are my data and what is the resolution? Acta Crystallogr D Biol Crystallogr 69, 1204-1214). Models for molecular replacement were created using SCULPTOR in the PHENIX software suite, using Fab from pdb:1hzh as a template (Adams, et al. (2010) PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr 66, 213-221). Molecular replacement was performed with PHASER (McCoy, et al. (2007) Phaser crystallographic software. J Appl Crystallogr 40, 658-674) using the Fab model generated with SCULPTOR and the interleukin-1 receptor accessory protein from pdb:4dep as a template. The structure was refined using Buster (Bricogne, et al. (2020) BUSTER version 2.11.7. in BUSTER version 2.11.7, 2.11.7 Ed., Global Phasing Ltd., Cambridge, United Kingdom) model building and least-squares fit analysis was performed with COOT (Emsley, et al. (2010) Features and development of Coot. Acta Crystallogr D Biol Crystallogr 66, 486-501). Figures 6A-C are modeled using PyMOL (Schrodinger, LLC.(2015) Created using the PyMOL Molecular Graphics System, Version 2.4.

[0237] [Table 33]

[0238] HDX MS Experimental Procedure: For HDX (hydrogen / deuterium exchange mass spectrometry) experiments, IL1RAP antigen was analyzed alone (control) and together with the anti-IL1RAP #A2 antibody of the present invention present at approximately equimolar ratio (mixed sample). All sample handling was performed by the LEAP HDX PAL system. To identify peptides, control samples were incubated with HO buffer (HO 10 mM sodium phosphate pH 7.4). For the exchange experiments, the control and mixed samples were incubated (in duplicate) with DO buffer (DO 10 mM sodium phosphate pH 7.4) at 10, 100, and 1000 seconds by the following procedure: (1) 4 μL of sample was added to 40 μL of H2O / DO buffer; (2) the mixture was incubated at 20 °C for various time points (0, 10, 100, and 1000 seconds); (3) 40 μL of incubated sample was transferred to 40 μL of quench buffer (4 M urea, 0.4 M TCEP-HCl) at 4 °C; (4) 60 μL of quenched sample was transferred to an immobilized protease XIII / pepsin column (1:1) by running mobile phase A (99% HO, 1% acetonitrile, and 0.1% formic acid) at 200 μL / min for 2 minutes. The digested peptides were desalted on a Vanguard Pre-column (ACQUITY UPLC BEH C18, 130 Å, 1.7 μm, 2.1 mm × 5 mm, NovaBioAssays) for 3 min and then separated by liquid chromatography using an Acquity UPLC BEH C18 Column 1.7 um, 1 mm × 50 mm (Waters) at a flow rate of 160 μl / min at 4 °C. The LC gradient solvent system consisted of mobile phase A (composition as above) and mobile phase B (0.1% formic acid, 5% H2O, and 95% acetonitrile). The percentage of mobile phase B was held at 5% for 5 min; increased from 5% to 15% at 5.6 min, to 40% at 10.4 min, to 90% at 11 min; held at 90% until 11.5 min; decreased to 5% at 12.4 min, then held at 5% until 14 min.After chromatographic separation, peptides were detected by a Thermo Scientific Orbitrap Fusion mass spectrometer operated in positive electrospray ionization mode. Data were then analyzed by Byonic software (Protein Metrics) to identify peptides and by HDExaminer software (Sierra Analytics) to calculate deuterium uptake.

[0239] result: The structure of anti-IL1RAP #A2 Fab in complex with the extracellular domain of IL-1RAcP (residues 21-368, Table 28) was determined. All residues of IL-1RAcP are visible in the electron density except for residues 21-23 and 351-368. The Fab chains could also be traced completely except for the four C-terminal residues of chain H and the first N-terminal residue of chain L. The crystal B-factors are lowest for the D3 domain and the Fab (86 Å2 and 82 Å2) and increase towards the N-terminus of IL-1RAcP (100 Å2 for domain D2 and 160 Å2 for domain D1). This may be explained by the crystal packing of the complex, where the D1 domain has the least number of contact points to neighboring molecules.

[0240] The structure of IL-1RAcP resembles the previously determined structures of human IL-1RAcP-IL-1RI-IL1β (Thomas et al. (2012) Structure of the activating IL-1 receptor signaling complex. Nature structural & molecular biology 19, 455-457) and IL-1RAcP-IL-1RII IL1β (Wang et al. (2010) Structural insights into the assembly and activation of IL-1beta with its receptors. Nat Immunol 11, 905-911), with rms deviations of all IL-1RAcP backbone Cα atoms of 2.7 Å and 2.4 Å, respectively. An exemplary anti-IL1RAP #A2 Fab of the invention binds to the D3 domain of IL-1RAcP, with the interface burying approximately 900 Å2 of the surface area on each binding partner. The epitope residues on the D3 domain largely coincide with residues involved in the IL1RAcP-IL-1RI interface. Although the interaction of IL1RAcP with IL-1RI extends to residues in the D2 and D3 domains of IL1RAcP, the interface of IL1RAcP and the anti-IL1RAP antibodies of the invention is restricted to the D3 domain.

[0241] Figures 5A-D illustrate the structural features of IL-1RAcP: anti-IL1RAP Ab of the invention and the IL-1RAcP-IL-1RI-IL1β complex in two different views related by a 90° rotation. In Figure 5A, IL-1RAcP is shown in light blue with a semi-transparent surface. The Fab is shown as a ribbon. The heavy and light chains are colored in dark and light gray, respectively. Figure 5B shows the location of the Fab on IL-1RAcP in gray. For clarity, the Fab is shown as a semi-transparent ribbon. Figure 5C shows the structure of the human IL-1RAcP-IL-1RI-IL1β ternary complex (pdb 4dep) for comparison, in the same orientation as Figure 5A. IL-1RI is colored in red and IL-1β in green. Figure 5D illustrates the IL-1RAcP side of the IL-1RAcP-IL-1RI interface (shown in orange). For clarity, IL-1RI and IL1β are shown as semi-transparent ribbons.

[0242] The epitope is formed by amino acid residues 238, 239, 241, 244-247, 249, 251-256, 261, 263, 265, 267, 269, 271, 301, 303, 305-307, 311, 313, and 315 as defined by the program PISA. This matches well with the epitope determined by HDX-MS (residues 226-262 and 269-273) demonstrated in Figures 6A-B, which compares the HDX and X-ray epitopes on IL1RAP. In Figure 6A, the HDX epitope is shown in yellow and includes residues 226-262 and 269-273. In Figure 6B, the X-ray epitope is shown (as also shown in Figure 6B, except that in this figure the anti-IL1RAP Fab of the invention has been removed for clarity).

[0243] The amino acids of the IL1RAP extracellular domain start at amino acid position 21 as shown in SEQ ID NO: 184 and extend to amino acid 367. IL1RAP domain 1 corresponds to amino acid positions 21-134 (SEQ ID NO: 185), domain 2 corresponds to amino acids 135-234 (SEQ ID NO: 186), and domain 3 is non-responsive to amino acid positions 235-267 (SEQ ID NO: 187). Domain 3 is further divided herein according to identified epitope binding regions defined by X-ray crystallography (XR-D3A / B corresponding to amino acid positions 235-315 (SEQ ID NO:188), XR-D3A corresponding to amino acid positions 235-273 (SEQ ID NO:189), and XR-D3B corresponding to amino acids 300-315 (SEQ ID NO:190)) and HDX mapping (HDX-D2 / 3 A corresponding to amino acid positions 226-262 (SEQ ID NO:191), and HDX-D2 / 3 B corresponding to amino acid positions 226-273 (SEQ ID NO:192)). The underlined and bolded amino acid positions in SEQ ID NOs: 202-207 correspond to identified contact amino acids K238, N239, V241, V244, I245, H246, S247, N249, H251, V252, V253, Y254, E255, K256, E261, L263, P265, T267, Y269, S271, S301, S303, S305, R306, T307, T311, T313, I315 of the epitope that binds exemplary anti-IL1RAP antibody #A2 by X-ray mapping.

[0244] IL1RAP ECD Accession Number: Q9NPH3 (amino acids 21 to 367) SERCDDWGLDTMRQIQVFEDEPARIKCPLFEHFLKFNYSTAHSAGLTLIWYWTRQDRDLEEPINFRLPENRISKEKDVLWFRPTLLNDTGNYTCMLRNTTYCSKVAFPLEVVQKDSCFNSPMKLPVHKLYIEYGIQRITCPNVDGYFPSSVKPTITWYMGCYKIQNFNNVIPEGMNLSFLIALISNNGNYTCVVTYPENGRTFHLTRTLTVKVVGSPKNAVPPVIHSPNDHVVYEKEPGEELLIPCTVYFSFLMDSRNEVWWTIDGKKPDDITIDVTINESISHSRTEDETRTQILSIKKVTSEDLKRSYVCHARSAKGEVAKAAKVKQKVPAPRYTVELACGFGAT (SEQ ID NO: 184) IL1RAP ECD domain 1 (amino acids 21-134): SERCDDWGLDTMRQIQVFEDEPARIKCPLFEHFLKFNYSTAHSAGLTLIWYWTRQDRDLEEPINFRLPENRISKEKDVLWFRPTLLNDTGNYTCMLRNTTYCSKVAFPLEVVQK (SEQ ID NO: 185)

[0245] IL1RAP ECD domain 2 (amino acids 135-234): DSCFNSPMKLPVHKLYIEYGIQRITCPNVDGYFPSSVKPTITWYMGCYKIQNFNNVIPEGMNLSFLIALISNNGNYTCVVTYPENGRTFHLTRTLTVKVV (SEQ ID NO: 186) IL1RAP ECD domain 3 (amino acids 235-367): GSP KN A V PP VIHS P N D HVVYEK EPG E E L LI P C T V Y F S FLMDSRNEVWWTIDGKKPDDITIDVTINE S I S HSRT EDE T R T Q I LSIKKVTSEDLKRSYVCHARSAKGEVAKAAKVKQKVPAPRYTVELACGFGAT (SEQ ID NO: 187) IL1RAP ECD XR-D3A / B (amino acids 235-315): GSP KN A V PP VIHS P N D HVVYEK EPG E E L LI P C T V Y F S FLMDSRNEVWWTIDGKKPDDITIDVTINE S I S H SRT EDE T R T Q I (SEQ ID NO:188)

[0246] IL1RAP ECD XR D3A (amino acids 235-273): GSP KN A V PP VIHS P N D HVVYEK EPG E E L LI P C T V Y F S FL (SEQ ID NO: 189) IL1RAP ECD XR D3B (amino acids 300-315): S I S H SRT EDE T R T Q I (SEQ ID NO:190) IL1RAP ECD HDX D2 / 3 A (amino acids 226-262): TRTLTVKVVGSP KN A V PP VIHS P N DHVVYEK EPG E E L (SEQ ID NO:191) IL1RAP ECD HDX D2 / 3 B (amino acids 226-273): TRTLTVKVVGSP KN A V PP VIHS P N D HVVYEK EPG E E L LI P C T V Y F S FL (SEQ ID NO: 192)

[0247] The paratope is formed by residues I28 and L30-W33 from CDR-H1, F52, A54, S55, S57, and Y59 from CDR-H2, K74 from the loop following CDR-H2, and Y102-G107 and Y109 from CDR-H3. From the light chain, only CDR-L2 (residues E56 and G57) and CDR-L3 (residues G93 and T94) are critical for binding IL1RAP. The polar interactions include 10 hydrogen bonds and one salt bridge. These are summarized in Table 29.

[0248] [Table 34] Two antibodies, CAN03 and CAN04, were reported to bind to domains 3 and 2, respectively, inhibit IL-1 signaling, and partially inhibit IL-33 signaling (see Wang et al., 2010, Nature Immunology, 11:905-912 and WO2020 / 035577). CAN04 and CAN03 were synthesized according to the VH and VL sequences described in WO2020 / 035577 (herein incorporated by reference) and used for comparison with an exemplary anti-IL1RAP antibody (#A2). Table 30 summarizes a comparison of CAN04 versus an exemplary anti-IL1RAP (#A2) chimeric precursor. The CAN04 Ab did not show as broad inhibition across multiple measured parameters as #A2 in the Hek-Blue and NCI-H292 assays, and only demonstrated comparable inhibitory potency against IL36γ in the WB (whole blood) assay.

[0249] For the CAN03 Ab, although antibody binding was detected, there was no measurable inhibition in our in vitro screening assay (data not shown). This was consistent with previous findings reported in WO2020 / 035577, where only when CAN03 was combined with CAN04 was measurable IL-1B inhibition detected at the level of CAN04 alone.

[0250] [Table 35]

[0251] The "CAN04" Ab was reported to bind to domain 2 of IL1RAP, i.e., within amino acids 135-234 of IL1RAP (see Wang et al., 2010, Nature Immunology, 11:905-912), and the epitope to which the CAN04 antibody binds was reported to be located within amino acids 135-154, 155-174, 175-194, 195-214 of IL1RAP, or between amino acids 215-234. The "CAN03" antibody was reported to bind to domain 3 of IL1RAP, which consists of the structural region defined by amino acids 235-369 of IL1RAP. However, when comparing the HDX measurements for the epitopes of #A2, and CAN04 and CAN03, their epitopes were completely different, even without overlapping amino acids. This may be due to the nonlinear or conformational nature of the diverse epitopes recognized by CAN04 and CAN03. This could also explain the difference in biological activity, especially of CAN03, which does not have inhibitory activity, when compared to the #A2 antibody.

[0252] Example 5 Measurement of soluble IL1RAP and cyno PK Soluble IL1RAP is known to be present in human and cynomolgus monkey blood. To determine how soluble IL1RAP may affect the PK of anti-IL1RAP antibodies in vivo, free and total anti-IL1RAP levels were quantified in cyno serum samples.

[0253] Determination of free anti-IL1RAP antibodies: Serum concentrations of free anti-IL1RAP antibodies were measured using an enzyme-linked immunosorbent assay (ELISA) method. Briefly, microtiter plates (Nunc) were coated with 2 μg / mL recombinant cynomolgus IL1RAP overnight at 2-8 °C. Unbound capture reagent was washed off (1x PBS with 0.05% Tween 20) and wells were blocked with 5% BSA (Seracare) and incubated for 1 h at room temperature. Plates were then washed and calibration standards, QCs, and sample serial dilutions were added and incubated for 1 h at room temperature. Plates were washed and horseradish peroxidase (HRP)-conjugated goat anti-human IgG (Southern Biotech) pre-adsorbed against monkey serum proteins was added and incubated for 1 h at room temperature. After washing, bound HRP conjugate was detected using tetramethylbenzidine (TMB) substrate. The reaction was stopped with 1M H2SO4 and absorbance was measured using a SpectraMax® microplate reader at dual wavelengths of 450 and 650 nm. The signal produced was proportional to the amount of anti-Ang1 antibody present in the sample. Softmax Pro software (v5.4) was used for calibration standard curve fitting using a four-parameter logistic model and back-calculation of all unknown sample concentrations. (See FIG. 7A.)

[0254] Determination of total anti-IL1RAP antibodies Serum concentrations of total anti-IL1RAP antibodies were measured using a homogenous Meso Scale Discovery (MSD) electrochemiluminescence assay. Briefly, gold small spot streptavidin plates (MSD part #L45SA) were blocked with 5% BSA (Seracare) for 1 hour. Plates were then washed and capture and detection master mix of biotin and MSD sulfo-tag conjugated goat anti-human IgG (Southern Biotech) pre-adsorbed to monkey serum proteins containing calibration standards, QCs, and sample serial dilutions were added and incubated for 2 hours at room temperature. After washing, 2x MSD Read Buffer was added and the electrochemiluminescence signal was then measured on a Sector Imager 6000 instrument. Unknown serum concentrations were calculated from a standard curve fitted to a four-parameter logistic equation using MSD Discovery Workbench software. (See Figure 7B.)

[0255] Determination of free and total soluble IL1RAP Free and total soluble IL1RAP serum concentrations were determined using a Gyrolab™ workstation using biotinylated and Alexa Fluor 647-labeled anti-IL1RAP antibodies as capture and detection reagents, respectively. Total soluble IL1RAP concentrations were measured using monoclonal anti-IL1RAP capture and detection antibodies that recognize distinct epitopes from the therapeutic mAb. Free soluble IL1RAP concentrations were measured using monoclonal anti-IL1RAP capture antibodies and therapeutic anti-IL1RAP as detection. Reference standards and study samples were diluted in 2% BSA buffer. Samples and reagents were placed in a 96-well polypropylene microplate and sealed with a microplate foil sealer before loading into the instrument. Addition of capture reagents, samples, standards, detection reagents, and wash solutions to the microstructures on the CD was a fully automated process. During the assay, the method components were robotically loaded onto streptavidin beads inside a microcolumn structure on the Bioaffy CD starting with biotinylated capture antibody, wash buffer, then calibration standards, and samples, wash buffer, then AlexaFluor 647-conjugated detection antibody. After washing to remove unbound Alexa Fluor, the fluorescence intensity was measured and was proportional to the amount of soluble IL1RAP present in the sample. Analyte concentrations were determined from data regression of fluorescence values ​​derived from Alexa647 fluorescence emission converted to concentration values. Quantification was based on a four-parameter logistic (1 / Y2) regression from a recombinant cynomolgus monkey standard curve. (See FIG. 7C.)

[0256] result: Binding to surface IL1RAP is required for blocking activity Although IL1RAP binds not only to surface IL1RAP but also to the soluble form of ILRAP, an effective dose could be predicted. array:

[0257] [Table 36-1] [Table 36-2]

Table 36-3

Table 36-4

Table 36-5

Table 36-6

Table 36-7

Table 36-8

Table 36-9

Table 36-10

Table 36-11

Table 36-12

Table 36-13

Table 36-14

Table 36-15

Claims

1. K equal to or <0.1 nM D An anti-IL1RAP antibody or antigen-binding fragment thereof that binds to human IL1RAP at

2. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 1 which is a monoclonal antibody or antigen-binding fragment thereof.

3. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 1 , which is a humanized antibody or antigen-binding fragment thereof.

4. K equal to or <200 pM D 4. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 3, which binds to human and cynomolgus monkey IL1RAP at 100-150 nm.

5. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 3, 6, 117, 118, 119, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, or 135 (L-CDR1); the amino acid sequence of SEQ ID NO: 4, 7, 136, 137, 138, 139, 140, or 141 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8, 11, 12, 14, 142, 143, 144, 145, 146, or 147 (H-CDR1); the amino acid sequence of SEQ ID NO: 9, 13, 15, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, or 167 (H-CDR2); or the amino acid sequence of SEQ ID NO: 10 or 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 1 .

6. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1 = M or I, and X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1 = M, I, X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 112 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (H-CDR2), where amino acid X1=D or G, and X2=T or A; and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

7. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1 = M or I, and X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1 = M, I, X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 7 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (H-CDR2), where amino acid X1=D or G, and X2=T or A; and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

8. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 146, where X1 = M or I, and X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10, or c) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and d) a heavy chain variable region comprising the amino acid sequence (H-CDR1) of SEQ ID NO: 147, where X1 = M, I, X2 = N or S; the amino acid sequence (H-CDR2) of SEQ ID NO: 165, where X1 = D or G; X2 = A or T; X3 = N or A; X4 = Q or E; X5 = M or K; X6 = Q or K; X7 = D or G; the amino acid sequence (H-CDR3) of SEQ ID NO: 10; or e) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 112 (H-CDR1); the amino acid sequence of SEQ ID NO: 166 (wherein amino acid X1=D or G) (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 127 (L-CDR1); the amino acid sequence of SEQ ID NO: 139 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 167 (H-CDR2), where amino acid X1=D or G, and X2=T or A; and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

9. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:9 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 9 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

10. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

11. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 13 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 15 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

12. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

13. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

14. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 162 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

15. a) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or c) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 144 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or e) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 134, where X1 = A or T; X2 = Q or E; X3 = S or N; X4 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 140, where X1 = K or S; X2 = S or T; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 (H-CDR1); the amino acid sequence of SEQ ID NO: 161 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or g) a light chain variable region comprising the amino acid sequence (L-CDR1) of SEQ ID NO: 135, where X1 = Q or E; X2 = S or N; X3 = N or S; the amino acid sequence (L-CDR2) of SEQ ID NO: 141, where X1 = K or S; the amino acid sequence (L-CDR3) of SEQ ID NO: 5; and h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 (H-CDR1); the amino acid sequence of SEQ ID NO: 163 (H-CDR2); and the amino acid sequence of SEQ ID NO: 16 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

16. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO:3 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); and the amino acid sequence of SEQ ID NO:5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:9 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

17. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

18. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

19. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

20. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:148 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

21. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:149 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

22. a) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137; and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3). The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 5 .

23. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 1, comprising: a light chain variable region comprising any one of the amino acid sequences of SEQ ID NOs: 17, 36, 40, 47, 50, 51, or 52; and a heavy chain variable region comprising any one of the amino acid sequences of SEQ ID NOs: 67, 86, 97, 100, 101, or 102.

24. A light chain variable region comprising the amino acid sequence of SEQ ID NO: 17; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 67; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:36; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:86; or A light chain variable region comprising the amino acid sequence of SEQ ID NO: 40; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 90; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:47; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:50; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:100; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:51; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:101; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:52; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:

102. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 23, comprising:

25. 2. The anti-IL1RAP antibody of claim 1, comprising a light chain comprising the amino acid sequence of any one of SEQ ID NOs: 170, 171, 172, 173, 174, 175, or 176; and a heavy chain comprising the amino acid sequence of any one of SEQ ID NOs: 177, 178, 179, 180, 181, 182, or 183.

26. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 170; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

177.

27. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 171; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

178.

28. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 172; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

179.

29. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 173; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

180.

30. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 174; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

181.

31. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 175; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

182.

32. 26. The anti-IL1RAP antibody of claim 25, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 176; and a heavy chain comprising the amino acid sequence of SEQ ID NO:

183.

33. a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 118 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 119 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 149 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 150 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 120 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 152 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 121 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 152 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 122 (L-CDR1); the amino acid sequence of SEQ ID NO: 138 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 154 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 155 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 121 (L-CDR1); the amino acid sequence of SEQ ID NO: 138 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 120 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 122 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 156 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 123 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 143 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 124 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 158 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 151 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 152 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 154 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 155 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 156 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 143 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 136 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 152 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 153 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 154 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 155 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 156 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 143 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 148 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 117 (L-CDR1); the amino acid sequence of SEQ ID NO: 137 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 157 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 125 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 159 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 125 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); and the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 142 (H-CDR1); the amino acid sequence of SEQ ID NO: 160 (H-CDR2); and the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO: 125 (L-CDR1); the amino acid sequence of SEQ ID NO: 4 (L-CDR2); the amino acid sequence of SEQ ID NO: 5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 (H-CDR1); the amino acid sequence of SEQ ID NO: 159 (H-CDR2); the amino acid sequence of SEQ ID NO: 10 (H-CDR3); or a light chain variable region comprising the amino acid sequence of SEQ ID NO:125 (L-CDR1); the amino acid sequence of SEQ ID NO:4 (L-CDR2); the amino acid sequence of SEQ ID NO:5 (L-CDR3); and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8 (H-CDR1); the amino acid sequence of SEQ ID NO:160 (H-CDR2); and the amino acid sequence of SEQ ID NO:10 (H-CDR3). The anti-IL-36R antibody or antigen-binding fragment thereof of claim 1,

34. A light chain variable region comprising the amino acid sequence of SEQ ID NO:20; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:70; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:21; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:71; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:22; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:72; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:23; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:73; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:24; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:74; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:25; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:75; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:26; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:76; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:27; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:28; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:78; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:29; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:79; or A light chain variable region comprising the amino acid sequence of SEQ ID NO: 30; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 80; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:31; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:81; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:32; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:82; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:33; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:83; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:34; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:84; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:35; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:85; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:37; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:87; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:38; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:88; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:39; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:89; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:41; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:91; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:42; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:92; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:43; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:93; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:44; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:94; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:45; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:95; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:46; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:96; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:48; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:98; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:49; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:99; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:53; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:103; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:54; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:104; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:55; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:105; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:56; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:106; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:57; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:107; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:58; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:108; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:59; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:109; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:60; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:110; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:61; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:111; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:62; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:112; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:63; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:113; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:64; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:114; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:65; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:115; or A light chain variable region comprising the amino acid sequence of SEQ ID NO:66; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:

116. The anti-IL1RAP antibody or antigen-binding fragment thereof of claim 33, comprising:

35. An anti-IL1RAP antibody or antigen-binding fragment according to any one of claims 1 to 34 for use as a medicament.

36. A pharmaceutical composition comprising the antibody or antigen-binding fragment of any one of claims 1 to 34 and a pharma- ceutically acceptable carrier.

37. 37. The pharmaceutical composition of claim 36 for use in a method for treating a disease selected from inflammatory diseases, autoimmune diseases, respiratory diseases, metabolic disorders, epithelial-mediated inflammatory disorders, fibrosis, and cancer.

38. 35. The anti-IL1RAP antibody or antigen-binding fragment of any one of claims 1 to 34, for use in treating a disease selected from an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, and cancer.

39. 35. Use of an anti-IL1RAP antibody or antigen-binding fragment of any one of claims 1 to 34 in the manufacture of a medicament for treating a disease selected from an inflammatory disease, an autoimmune disease, a respiratory disease, a metabolic disorder, an epithelial-mediated inflammatory disorder, fibrosis, and cancer.

40. 40. Use of the pharmaceutical composition of claim 37, the anti-IL1RAP antibody or antigen-binding fragment of claim 38, or the anti-IL1RAP antibody or antigen-binding fragment of claim 39, wherein the disease is selected from psoriasis, psoriatic arthritis, multiple sclerosis, rheumatoid arthritis, COPD, chronic asthma, and ankylosing spondylitis.

41. 2. The isolated polynucleotide of claim 1, which encodes a sequence defined by one or more of SEQ ID NOs: 1-167 or 170-183.

42. 42. A vector comprising a polynucleotide according to claim 41, preferably an expression vector, more preferably a vector comprising a polynucleotide according to the invention operably linked to an expression control sequence.

43. 43. A host cell comprising the polynucleotide and vector of claim 42.

44. 35. A method for producing an anti-IL1RAP antibody or antigen-binding fragment of any one of claims 1 to 34, comprising the steps of: (a) culturing a host cell under conditions allowing expression of the anti-IL1RAP antibody or antigen-binding fragment; and (b) recovering the anti-IL1RAP antibody or antigen-binding fragment.

45. A diagnostic kit comprising the anti-IL1RAP antibody or antigen-binding fragment of any one of claims 1 to 34.

46. 46. ​​The diagnostic kit of claim 45 for the diagnosis of inflammatory diseases, autoimmune diseases, respiratory diseases, metabolic disorders, epithelial-mediated inflammatory disorders, fibrosis, cancer, psoriasis, psoriatic arthritis, multiple sclerosis, rheumatoid arthritis, COPD, chronic asthma, or ankylosing spondylitis.

47. 35. An isolated antibody or antigen-binding fragment that binds to the following residues K238, N239, V241, V244, I245, H246, S247, N249, H251, V252, V253, Y254, E255, K256, E261, L263, P265, T267, Y269, and S271 when bound to IL1RAP, and blocks binding of any of the antibodies of claims 1-34.

48. 48. The isolated monoclonal antibody of claim 47, which blocks binding of anti-IL1RAP to IL1RAP by at least 80%.

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