US2020299349A1PendingUtilityA1

Partial agonists of interleukin-2

48
Assignee: UNIV LELAND STANFORD JUNIORPriority: Nov 21, 2017Filed: Nov 20, 2018Published: Sep 24, 2020
Est. expiryNov 21, 2037(~11.3 yrs left)· nominal 20-yr term from priority
Inventors:Kenan Christopher GarciaSonia S. MajriCaleb R. GlassmanLeon Lih-Ren Su
C07K 14/55C12N 15/09C07K 2319/30C07K 2319/31A61K 38/00A61K 9/0024C07K 2317/74A61P 37/00C07K 16/244C07K 2317/92C12N 15/63
48
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Claims

Abstract

Provided herein, inter alia, are human interleukin-2 (IL-2) muteins or variants thereof. In particular, provided herein are IL-2 muteins that have a decreased binding capacity for IL-2Rγ. Such IL-2 muteins are useful, for example, as IL-2 partial agonist in applications where reduction or inhibition of one or more IL-2 and/or IL-15 functions is useful (e.g., in the treatment of autoimmune diseases or conditions). Also provided are nucleic acids encoding such IL-2 muteins, methods of making such IL-2 muteins, pharmaceutical compositions that include such IL-2 muteins and methods of treatment using such pharmaceutical compositions.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . An interleukin 2 (IL-2) mutein having:
 (a) reduced binding affinity for interleukin 2 receptor γ (IL-2Rγ) as compared to an IL-2 polypeptide encoded by SEQ ID NO: 2; and   (b) 15-95% E max  as compared to an IL-2 polypeptide encoded by SEQ ID NO: 2.   
     
     
         2 . The IL-2 mutein of  claim 1 , wherein the mutein comprises:
 (i) one or more amino acid substitutions that increase IL-2Rβ binding affinity compared to a polypeptide encoded by SEQ ID NO: 1, selected from L80F, R81D, L85V, I86V, and I92F, numbered in accordance with the amino acid sequence of SEQ ID NO: 1; and/or   (ii) one or more amino acid substitutions that reduce IL-2Rγ receptor binding affinity and results in 15-95% of E mx  compared to a polypeptide encoded by SEQ ID NO: 2, selected from (A) L18R and Q22E; and (B) the amino acid position 126, numbered in accordance with the amino acid sequence of SEQ ID NO: 2.   
     
     
         3 . The IL-2 mutein of  claim 2 , wherein the amino acid substitution at position 126 of SEQ ID NO: 2 is selected from the group consisting of Q126A, Q126C, Q126D, Q126E, Q126G, Q126H, Q126I, Q126K, Q126M, Q126R, Q126S, or Q126T. 
     
     
         4 . The IL-2 mutein of  claim 3 , wherein the mutein comprises the amino acid substitution Q126H, Q126K, or Q126M. 
     
     
         5 . The IL-2 mutein of  claim 3 , wherein the mutein comprises an amino acid substitution Q126H. 
     
     
         6 . An interleukin 2 (IL-2) mutein having (a) reduced binding affinity for interleukin 2 receptor γ (IL-2Rγ); and (b) 15-95% E max  as compared to a polypeptide encoded by SEQ ID NO: 1. 
     
     
         7 . The IL-2 mutein of  claim 6 , wherein the mutein comprises one or more amino acid substitutions that reduce IL-2Rγ receptor binding affinity and results in 15-95% E max  as compared to a polypeptide encoded by SEQ ID NO: 1, selected from (A) L18R and Q22E; and (B) amino acid position 126, numbered in accordance with the amino acid sequence of SEQ ID NO: 1. 
     
     
         8 . The IL-2 mutein of  claim 7 , wherein the mutein comprises an amino acid substitution at position 126 of SEQ ID NO: 1 selected from the group consisting of Q126A, Q126C, Q126D, Q126E, Q126G, Q126H, Q126I, Q126K, Q126M, Q126R, Q126S, or Q126T. 
     
     
         9 . The IL-2 mutein of  claim 8 , wherein the mutein comprises an amino acid substitution Q126H, Q126K, or Q126M. 
     
     
         10 . The IL-2 mutein of  claim 9 , wherein the mutein comprises an amino acid substitution Q126H. 
     
     
         11 . The IL-2 mutein of any one of  claims 1 - 10 , wherein the mutein is structurally modified to increase half-life. 
     
     
         12 . The IL-2 mutein of  claim 11 , wherein said modification comprises one or more modifications selected from the group consisting of fusion to a human Fc antibody fragment, fusion to albumin, and PEGylation. 
     
     
         13 . The IL-2 mutein of any one of  claims 1 - 12 , wherein said mutein increases proliferation of regulatory T (T reg ) cells and/or induces minimal proliferation of potentially inflammatory T cells. 
     
     
         14 . The IL-2 mutein of any one of  claims 1 - 13 , wherein said mutein causes expansion of T reg  cells and does not promote expansion of potentially inflammatory T cells and Natural Killer (NK) cell or granulocytes. 
     
     
         15 . The IL-2 mutein of  claim 14 , wherein the mutein induces less proliferation of potentially inflammatory T cells compared to the polypeptide encoded by SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         16 . The IL-2 mutein of  claim 15 , wherein said potentially inflammatory T cells are CD4 +  IFNγ +  T cells or are CD8 + IFNγ +  T cells. 
     
     
         17 . The IL-2 mutein of any one of  claims 1 - 16 , wherein the mutein increases proliferation of T reg  cells by at least 3 fold and/or induces less IFNγ secretion compared to the polypeptide encoded by SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         18 . The IL-2 mutein of any one of  claims 1 - 17 , wherein said mutein does not induce IFNγ secretion from CD8 +  T cells and/or other inflammatory immune cell subsets. 
     
     
         19 . The IL-2 mutein of any one of  claims 1 - 18 , wherein the mutein has 70-95% E max  as compared to the polypeptide encoded by SEQ ID NO: 1. 
     
     
         20 . A nucleic acid encoding the IL-2 mutein of any one of  claims 1 - 19 . 
     
     
         21 . A vector comprising the nucleic acid of  claim 20 . 
     
     
         22 . A host cell comprising the nucleic acid of  claim 20  or the vector of  claim 21 . 
     
     
         23 . A sterile pharmaceutical composition comprising the IL-2 mutein of any one of  claims 1 - 19  and a pharmaceutically acceptable excipient. 
     
     
         24 . A syringe comprising (a) the IL-2 mutein of any one of  claims 1 - 19 ; (b) the nucleic acid of  claim 20 ; (c) the vector of  claim 21 ; (d) the host cell of  claim 22 ; and/or (e) the pharmaceutical composition of  claim 23 . 
     
     
         25 . A catheter comprising (a) the IL-2 mutein of any one of  claims 1 - 19 ; (b) the nucleic acid of  claim 20 ; (c) the vector of  claim 21 ; (d) the host cell of  claim 22 ; and/or (e) the pharmaceutical composition of  claim 23 . 
     
     
         26 . A kit comprising:
 (a) one or more of (i) the IL-2 mutein of any one of  claims 1 - 19 ; (ii) the nucleic acid of  claim 20 ; (iii) the vector of  claim 21 ; (iv) the host cell of  claim 22 ; (v) the sterile pharmaceutical composition of  claim 23 ; (vi) the syringe of  claim 24 ; and/or (vii) the catheter of  claim 25 ; and   (b) written instructions for using the (i) the IL-2 mutein of any one of  claims 1 - 19 ; (ii) the nucleic acid of  claim 20 ; (iii) the vector of  claim 21 ; (iv) the host cell of  claim 22 ; (v) the sterile pharmaceutical composition of  claim 23 ; (vi) the syringe of  claim 24 ; and/or (vii) the catheter of  claim 25 .   
     
     
         27 . A method for treating an autoimmune disease in an individual in need thereof comprising administering (a) a therapeutically effective amount of the IL-2 mutein of any one of  claims 1 - 19 ; (b) the nucleic acid of  claim 20 ; (c) the vector of  claim 21 ; (d) the host cell of  claim 22 ; and/or (e) the pharmaceutical composition of  claim 23  to the individual. 
     
     
         28 . The method of  claim 27 , wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, insulin-dependent diabetes mellitus, hemolytic anemias, rheumatic fever, thyroiditis, Crohn's disease, myasthenia gravis, glomerulonephritis, autoimmune hepatitis, multiple sclerosis, alopecia areata, psoriasis, vitiligo, dystrophic epidermolysis bullosa, systemic lupus erythematosus, and graft vs. host disease. 
     
     
         29 . The method of  claim 28 , wherein the autoimmune disease is graft vs. host disease. 
     
     
         30 . The method of any one of  claims 27 - 29 , further comprising administering the IL-2 mutein of any one of  claims 1 - 19  or the pharmaceutical composition of  claim 23  in combination with an antibody that targets the mutein to a specific cell type. 
     
     
         31 . The method of  claim 30 , wherein the cell type is a regulatory T (T reg ) cell. 
     
     
         32 . The method of  claim 30  or  claim 31 , wherein the antibody is covalently or non-covalently linked to the IL-2 mutein. 
     
     
         33 . A method for producing the IL-2 mutein of any one of  claims 1 - 19  comprising culturing the host cell of  claim 22  under suitable conditions for the production of the mutein. 
     
     
         34 . The method of  claim 33  further comprising isolating and/or purifying the produced IL-2 mutein. 
     
     
         35 . The method of  claim 33  or  34 , further comprising structurally modifying the IL-2 mutein to increase half-life. 
     
     
         36 . The method of  claim 35 , wherein said modification comprises one or more alterations selected from the group consisting of fusion to a human Fc antibody fragment, fusion to albumin, and PEGylation. 
     
     
         37 . A method for preventing the proliferation of potentially inflammatory T cells and/or preventing secretion of IFNγ from CD8 +  T cells or other inflammatory immune cell subsets, said method comprising contacting a cell expressing an interleukin 2 receptor γ (IL-2Rγ) with the IL-2 mutein of any one of  claims 1 - 19 . 
     
     
         38 . The method of  claim 37 , wherein said potentially inflammatory T cells are CD4 + CD44 + IFNγ +  T cells or are CD8 + CD44 + IFNγ +  T cells. 
     
     
         39 . The method of  claim 37  or  claim 38 , wherein the method is performed in vitro, in vivo, or ex vivo. 
     
     
         40 . A method for decreasing proliferation of regulatory T (T reg ) cells comprising contacting a T reg  cell with an interleukin 2 (IL-2) mutein having: (i) reduced binding affinity for interleukin 2 receptor γ (IL-2Rγ) as compared to the polypeptide encoded by SEQ ID NO: 2; and (ii) 0-50% E max  as compared to the polypeptide encoded by SEQ ID NO: 2. 
     
     
         41 . The method of  claim 40 , wherein the IL-2 mutein comprises:
 (i) one or more amino acid substitutions that increase IL-2Rβ binding affinity compared to the polypeptide encoded by SEQ ID NO: 1, selected from L80F, R81D, L85V, I86V, and I92F, numbered in accordance with the amino acid sequence of SEQ ID NO: 1; and   (ii) one or more amino acid substitutions that reduce IL-2Rγ receptor binding affinity and results in 0-50% of the E max  compared to the polypeptide encoded by SEQ ID NO: 2, selected from (A) L18R and Q22E; and (B) the amino acid position 126, numbered in accordance with the amino acid sequence of SEQ ID NO: 2.   
     
     
         42 . A method for decreasing proliferation of regulatory T (T reg ) cells comprising contacting a T reg  cell with an IL-2 mutein having: (i) reduced binding affinity for IL-2Rγ as compared to the polypeptide encoded by SEQ ID NO: 1; and (ii) 0-50% E max  as compared to the polypeptide encoded by SEQ ID NO: 1. 
     
     
         43 . The method of  claim 42 , wherein the mutein comprises one or more amino acid substitutions that reduce IL-2Rγ receptor binding affinity and results in 0-50% E max  as compared to a polypeptide encoded by SEQ ID NO: 1, selected from (A) L18R and Q22E; and (B) amino acid position 126, numbered in accordance with the amino acid sequence of SEQ ID NO: 1. 
     
     
         44 . The method of  claim 41  or  claim 43 , wherein the amino acid substitution at position 126 of SEQ ID NO: 1 or SEQ ID NO: 2 is selected from the group consisting of Q126A, Q126C, Q126D, Q126E, Q126G, Q126H, Q126I, Q126K, Q126M, Q126R, Q126S, or Q126T. 
     
     
         45 . The method of  claim 44 , wherein the mutein comprises the amino acid substitution Q126H, Q126K, or Q126M. 
     
     
         46 . The method of  claim 45 , wherein the mutein comprises an amino acid substitution Q126H. 
     
     
         47 . The method of any one of  claims 39 - 46 , further comprising administering the muteins with one or more antibodies that target the mutein to a T reg  cell. 
     
     
         48 . The method of  claim 47 , wherein the antibody is covalently or non-covalently linked to the mutein. 
     
     
         49 . The method of any one of  claims 39 - 48 , wherein the method is performed in vitro, in vivo, or ex vivo.

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