# US 12,178,847 B2 · Compositions and methods for adjoining type I and type II extracellular domains as heterologous chimeric proteins > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,178,847 B2 - Document ID: US12178847B2 - Status: Granted - Kind: B2 - Application number: 16/813,165 - Filed: 2020-03-09 - Published: 2020-08-06 - Granted: 2024-12-31 - Earliest claimed: 2015-10-01 - Assignees: Kopfkino IP, LLC - Applicants: Kopfkino IP, LLC - Inventors: Taylor Schreiber; George Fromm; Suresh De Silva; Neal Schilling - Web page: https://patentgrep.com/patent/US12178847B2 ## CPC - A61K 38/1774: {Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)} - A61P 35/00: Antineoplastic agents - A61K 38/177: {Receptors; Cell surface antigens; Cell surface determinants} - A61P 37/02: Immunomodulators - A61P 37/04: Immunostimulants - A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} - 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 2319/00: Fusion polypeptide - C07K 2319/74: containing a fusion for binding to a cell surface receptor - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract The present invention relates to, inter alia, compositions and methods, including chimeric proteins that find use in the treatment of disease, such as immunotherapies for cancer and autoimmunity. In part, the invention provides, in various embodiments, fusions of extracellular domains of transmembrane proteins that can have stimulatory or inhibitory effects. ## Claims ### Claim 1 A heterologous chimeric protein comprising: (a) a first domain comprising an extracellular domain of human programmed cell death protein 1 (PD-1) that is capable of binding a PD-1 ligand, and comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 32; (b) a second domain comprising an extracellular domain of human glucocorticoid-induced TNFR-related (GITR) ligand (GITRL) that is capable of binding a GITRL receptor; and (c) a linker linking the first domain and the second domain and comprising a hinge-CH2-CH3Fc domain, wherein the linker comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 71 or SEQ ID NO: 72. ### Claim 2 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of inhibiting an immunosuppressive signal. ### Claim 3 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of: (a) reducing or eliminating an immune inhibitory signal when the portion of PD-1 is bound to its ligand and/or (b) increasing or activating an immune stimulatory signal when the portion of GITRL is bound to its receptor. ### Claim 4 (depends on 1) The heterologous chimeric protein of claim 1, wherein the PD-1 ligand is PD-L1. ### Claim 5 (depends on 1) The heterologous chimeric protein of claim 1, wherein the GITRL receptor is GITR. ### Claim 6 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of simultaneously binding the PD-1 ligand and the GITRL receptor, wherein the PD-1 ligand is PD-L1, and the GITRL receptor is GITR. ### Claim 7 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of reducing the ability of CD4+FoxP3+regulatory T cells to suppress the proliferation of conventional CD4+or CD8+T cells. ### Claim 8 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of enhancing tumor killing activity by T cells. ### Claim 9 (depends on 1) The heterologous chimeric protein of claim 1, wherein the heterologous chimeric protein is capable of providing a sustained immunomodulatory effect. ### Claim 10 (depends on 1) The heterologous chimeric protein of claim 1, wherein the linker further comprises one or more joining linkers having amino acid sequences selected from SEQ ID NOs: 23-31 and 73-78. ### Claim 11 (depends on 1) The heterologous chimeric protein of claim 1, wherein the chimeric protein is expressed by a mammalian host cell as a secretable and functional single polypeptide chain. ### Claim 12 (depends on 1) The heterologous chimeric protein of claim 1, wherein the portion of PD-1is at least 98% identical to the amino acid sequence of SEQ ID NO: 32. ### Claim 13 (depends on 1) The heterologous chimeric protein of claim 1, wherein the linker comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 71 or SEQ ID NO: 72. ### Claim 14 (depends on 1) An expression vector, comprising a nucleic acid encoding the heterologous chimeric protein of claim 1. ### Claim 15 (depends on 14) A host cell, comprising the expression vector of claim 14. ### Claim 16 (depends on 1) A pharmaceutical composition, comprising a therapeutically effective amount of the heterologous chimeric protein of claim 1. ### Claim 17 A recombinant fusion protein comprising: (a) a first domain comprising an extracellular domain of human programmed cell death 1 (PD-1) that is at least 95% identical to the amino acid sequence of SEQ ID NO: 32 and is capable of binding a PD-1 ligand, (b) a second domain comprising an extracellular domain of human glucocorticoid-induced TNFR-related (GITR) ligand (GITRL) and is capable of binding a GITRL receptor, and (c) a linker linking the first domain and the second domain, wherein the linker comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 71 or SEQ ID NO: 72. ### Claim 18 (depends on 1) A nucleic acid encoding the heterologous chimeric protein of claim 1.