Interleukin-18 variants and methods of use
The present invention provides compositions and methods comprising an activator of interleukin-18 (IL-18) activity for use in therapeutic and non-therapeutic applications. The activator provides IL-18 signaling activity even in the presence of an inhibitory molecule 5 such as IL-18 binding protein (IL-18BP).
1 . A method comprising administering to a subject a composition comprising a modified interleukin 18 (IL-18) polypeptide conjugated, with or without a linker, to an anti-PD-1 antibody, wherein the modified IL-18 polypeptide comprises an amino acid sequence having 93% or more sequence identity with the wild-type (WT) IL-18 sequence as set forth in SEQ ID NO: 30 and at least three mutations, relative to WT human IL-18 as set forth in SEQ ID NO:30, that reduce binding to IL-18 binding protein (IL-18BP) as compared to WT IL-18 as set forth in SEQ ID NO: 30, wherein the at least three mutations comprise at least one substitution at a position selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
2 . The method of claim 1 , wherein the at least three mutations comprise a substitution at Tyrosine-1 and the at least one substitution at a position selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
3 . The method of claim 1 , wherein the at least three mutations comprise at least two substitutions at positions selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
4 . The method of claim 1 , wherein the modified IL-18 polypeptide further comprises a substitution at Glutamic acid-6.
5 . The method of claim 1 , wherein the at least three mutations comprise the at least one substitution at a position selected from: Methionie-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionin-60, and at least one substitution at a position selected from: Glutamine-103, Serine-105, Aspartic acid-110, Asparagine-111, and Methionine-113.
6 . The method of claim 1 , wherein the subject has cancer.
7 . The method of claim 6 , wherein the subject has leukemia.
8 . The method of claim 6 , wherein the subject has multiple myeloma.
9 . The method of claim 1 , wherein the modified IL-18 polypeptide has an EC 50 for IL-18BP that is at least 10-fold higher than the EC 50 of WT IL-18 for IL-18BP.
10 . The method of claim 1 , wherein the modified IL-18 polypeptide has an IL-18 BP to IL-18 Receptor (IL-18R) dissociation constant ratio that is at least 2-fold higher than the IL-18 to IL-18R dissociation constant ratio of WT IL-18.
11 . The method of claim 10 , wherein the modified IL-18 polypeptide has an IL-18 BP to IL-18R dissociation constant ratio that is at least 20-fold higher than the IL-18 to IL-18R dissociation constant ratio of WT IL-18.
12 . The method of claim 1 , wherein the modified IL-18 polypeptide comprises at least four mutations, relative to WT IL-18 as set forth in SEQ ID NO: 30, that reduce binding to IL-18BP as compared to WT IL-18 as set forth in SEQ ID NO: 30.
13 . The method of claim 1 , wherein the modified IL-18 polypeptide comprises at least five mutations, relative to WT IL-18 as set forth in SEQ ID NO: 30, that reduce binding to IL-18BP as compared to WT IL-18 as set forth in SEQ ID NO: 30.
14 . The method of claim 6 , wherein the subject has an MHC class I deficient cancer.
15 . The method of claim 6 , wherein the subject has a solid tumor.
16 . The method of claim 1 , wherein the modified IL-18 polypeptide has a K D for IL-18BP of 10 nM or greater.
17 . The method of claim 16 , wherein the modified IL-18 variant polypeptide binds human IL-18Ra with an affinity that is at least comparable to WT human IL-18.
18 . A method comprising administering to a subject a composition comprising a nucleic acid encoding a modified interleukin 18 (IL-18) polypeptide fused, with or without a linker, to an anti-PD-1 antibody, wherein the modified IL-18 polypeptide comprises an amino acid sequence having 93% or more sequence identity with the wild-type (WT) IL-18 sequence as set forth in SEQ ID NO: 30 and at least three mutations, relative to WT human IL-18 as set forth in SEQ ID NO:30, that reduce binding to IL-18 binding protein (IL-18BP) as compared to WT IL-18 as set forth in SEQ ID NO: 30, wherein the at least three mutations comprise at least one substitution at a position selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
19 . The method of claim 18 , wherein the at least three mutations comprise a substitution at Tyrosine-1 and the at least one substitution at a position selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
20 . The method of claim 18 , wherein the at least three mutations comprise at least two substitutions at positions selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60.
21 . The method of claim 18 , wherein the modified IL-18 polypeptide further comprises a substitution at Glutamic acid-6.
22 . The method of claim 18 , wherein the at least three mutations comprise the at least one substitution at a position selected from: Methionine-51, Lysine-53, Serine-55, Glutamine-56, Proline-57, Glycine-59, and Methionine-60, and at least one substitution at Glutamine-103, Serine-105, Aspartic acid-110, Asparagine-111, or Methionine-113.
23 . The method of claim 18 , wherein the modified IL-18 polypeptide has an EC 50 for IL-18BP that is at least 10-fold higher than the EC 50 of WT IL-18 for IL-18BP.
24 . The method of claim 18 , wherein the modified IL-18 polypeptide has an IL-18 BP to IL-18 Receptor (IL-18R) dissociation constant ratio that is at least 2-fold higher than the IL-18 to IL-18R dissociation constant ratio of WT IL-18.
25 . The method of claim 24 , wherein the modified IL-18 polypeptide has an IL-18 BP to IL-18R dissociation constant ratio that is at least 20-fold higher than the IL-18 to IL-18R dissociation constant ratio of WT IL-18.
26 . The method of claim 18 , wherein the modified IL-18 polypeptide comprises at least four mutations, relative to WT IL-18 as set forth in SEQ ID NO: 30, that reduce binding to IL-18BP as compared to WT IL-18 as set forth in SEQ ID NO: 30.
27 . The method of claim 18 , wherein the modified IL-18 polypeptide comprises at least five mutations, relative to WT IL-18 as set forth in SEQ ID NO: 30, that reduce binding to IL-18BP as compared to WT IL-18 as set forth in SEQ ID NO: 30.
28 . The method of claim 18 , wherein the subject has cancer.