# US 12,006,347 B2 · Superagonists and antagonists of interleukin-2 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,006,347 B2 - Document ID: US12006347B2 - Status: Granted - Kind: B2 - Application number: 17/470,246 - Filed: 2021-09-09 - Published: 2022-06-30 - Granted: 2024-06-11 - Earliest claimed: 2010-12-22 - Assignees: The Board of Trustees of the Leland Stanford Junior University - Applicants: The Board of Trustees of the Leland Stanford Junior University - Inventors: Kenan Christopher Garcia; Aron Levin; Aaron Ring - Web page: https://patentgrep.com/patent/US12006347B2 ## CPC - C07K 14/55: IL-2 - A61P 35/00: Antineoplastic agents - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract Novel human interleukin-2 (IL-2) muteins or variants thereof, and nucleic acid molecules and variants thereof are provided. Methods for producing these muteins as well as methods for stimulating the immune system of an animal are also disclosed. In addition, the invention provides recombinant expression vectors comprising the nucleic acid molecules of this invention and host cells into which expression vectors have been introduced. Pharmaceutical compositions are included comprising a therapeutically effective amount of a human IL-2 mutein of the invention and a pharmaceutically acceptable carrier. The IL-2 muteins can be used in pharmaceutical compositions for use in treatment of cancer and in stimulating the immune response. ## Claims ### Claim 1 A nucleic acid encoding an IL-2Rβ binding protein, wherein the equilibrium dissociation constant for IL-2Rβ of the IL-2Rβ binding protein is less than that of wild-type human IL-2 (hIL-2), and wherein said IL-2Rβ binding protein is an IL-2 mutein comprising amino acid substitutions at positions L80F, R81D, L85V, I86V, and I92F numbered in accordance with wild-type hIL-2. ### Claim 2 (depends on 1) The nucleic acid of claim 1, wherein the binding protein further comprises the following amino acid substitution: F42A. ### Claim 3 (depends on 2) The nucleic acid of claim 2, wherein the binding protein further comprises one or more amino acid substitutions at positions selected from the group consisting of I24V, P65H, Q74R, Q74H, Q74N, Q74S, I89V, and V931. ### Claim 4 (depends on 3) The nucleic acid of claim 3, wherein position 74 is arginine. ### Claim 5 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: Q74N, L80F, R81D, L85V, I86V, I89V, and I92F. ### Claim 6 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: L80F, R81D, L85V, I86V, I89V, I92F, and V931. ### Claim 7 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: Q74S, L80F, R81D, L85V, I86V, and I92F. ### Claim 8 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: Q74H, L80F, R81D, L85V, I86V, and I92F. ### Claim 9 (depends on 1) The nucleic acid of claim 1, wherein the binding protein inhibits the interaction between the IL- 2Rβ and the IL-2Rγ. ### Claim 10 (depends on 9) The nucleic acid of claim 9, wherein the binding protein further comprises one or more amino acid substitutions at positions 18, 22, 24, 65, 74, 89, 93, and 126 numbered in accordance with wild-type hIL-2. ### Claim 11 (depends on 10) The nucleic acid of claim 10, wherein the binding protein has the following substitutions: L18R, Q22E, L80F, R81D, L85V, I86V, I89V, I92F, V931, and Q126T. ### Claim 12 (depends on 10) The nucleic acid of claim 10, wherein the binding protein has the following substitutions: L18R, Q22E, Q74S, L80F, R81D, L85V, I86V, I89V, I92F, V931, and Q126T. ### Claim 13 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: L18R, Q22E, L80F, R81D, L85V, I86V, I89V, I92F, V931, and Q126T. ### Claim 14 (depends on 1) The nucleic acid of claim 1, wherein the binding protein has the following substitutions: Q74N, L80F, R81D, L85V, I86V, and I92F. ### Claim 15 (depends on 1) The nucleic acid of claim 1, wherein the binding protein further comprises one or more amino acid substitutions at positions selected from the group consisting of 124, F42, K43, P65, Q74, I89, and V93. ### Claim 16 (depends on 1) The nucleic acid of claim 1, wherein the binding protein further comprises one or more amino acid substitutions at positions selected from the group consisting of F42 and K43. ### Claim 17 (depends on 1) The nucleic acid of claim 1, wherein the binding protein further comprises the following amino acid substitution: K43N. ### Claim 18 (depends on 1) The nucleic acid of claim 1, wherein the binding protein comprises the following substitutions: F42A, L80F, R81D, L85V, I86V, I89V, and I92F. ### Claim 19 (depends on 1) The nucleic acid of claim 1, wherein the binding protein comprises the following amino acid substitution: Q74N. ### Claim 20 (depends on 16) The nucleic acid of claim 16, wherein the binding protein exhibits reduced binding to an IL-2Rα relative to wild-type IL-2. ### Claim 21 (depends on 16) The nucleic acid of claim 16, wherein the binding protein comprises the following substitutions: L18R, Q22E, L80F, R81D, L85V, I86V, I89V, I92F, V931, and Q126T. ### Claim 22 (depends on 21) The nucleic acid of claim 21, wherein the binding protein exhibits reduced binding to IL-2Rγ relative to wild-type IL-2. ### Claim 23 (depends on 21) The nucleic acid of claim 21, wherein the binding protein exhibits reduced signaling through IL-2Rγ relative to wild-type IL-2. ### Claim 24 (depends on 1) The nucleic acid of claim 1, wherein the binding protein is fused to a heterologous polypeptide. ### Claim 25 (depends on 24) The nucleic acid of claim 24, wherein the heterologous polypeptide is an antibody Fc region and/or human serum albumin. ### Claim 26 (depends on 1) A vector comprising the nucleic acid sequence of claim 1. ### Claim 27 (depends on 1) The nucleic acid of claim 1, wherein the IL-2 mutein comprises the amino acid sequence of SEQ ID NO: 12.