Multivalent and multispecific gitr-binding fusion proteins

Abstract

This disclosure generally provides molecules that specifically engage glucocorticoid-induced TNFR-related protein (GITR), a member of the TNF receptor superfamily (TNFRSF). More specifically, the disclosure relates to multivalent and/or multispecific molecules that bind at least GITR.

What is claimed is: 
     
         1 . A polypeptide comprising at least two copies of a GITR-binding domain (GITR-BD), wherein the GITR-BD comprises a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 138; a complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO: 141; and a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 116, and wherein the polypeptide does not comprise a binding domain that binds to an antigen other than GITR. 
     
     
         2 . The polypeptide of  claim 1 , wherein the GITR-BD comprises the amino acid sequence of SEQ ID NO: 59. 
     
     
         3 . The polypeptide of  claim 2 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 93. 
     
     
         4 . A tetravalent GITR-targeting molecule comprising two copies of a fusion protein comprising the structure: (GITR-BD)-Linker-(GITR-BD)-Linker-Hinge-Fc, wherein
 (a) each GITR-BD is a GITR-binding domain comprising a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 138; a complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO: 141; and a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 116;   (b) Linker is a linker polypeptide;   (c) Hinge is a polypeptide derived from an immunoglobulin hinge region; and   (d) Fc is an immunoglobulin Fc region polypeptide;   
       and wherein the tetravalent GITR-targeting molecule does not comprise a binding domain that binds to an antigen other than GITR. 
     
     
         5 . The tetravalent GITR-targeting molecule of  claim 4 , wherein the Fc comprises an amino acid sequence selected from SEQ ID NO: 1-6. 
     
     
         6 . The tetravalent GITR-targeting molecule of  claim 5 , wherein the Hinge comprises the amino acid sequence of SEQ ID NO: 8. 
     
     
         7 . The tetravalent GITR-targeting molecule of  claim 6 , wherein each Linker comprises an amino acid sequence independently selected from SEQ ID NO: 12 and SEQ ID NO: 16. 
     
     
         8 . The tetravalent GITR-targeting molecule of  claim 4 , wherein each GITR-BD comprises the amino acid sequence of SEQ ID NO: 59. 
     
     
         9 . The tetravalent GITR-targeting molecule of  claim 8 , wherein the Fc comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         10 . The tetravalent GITR-targeting molecule of  claim 9 , wherein the Hinge comprises the amino acid sequence of SEQ ID NO: 8. 
     
     
         11 . The tetravalent GITR-targeting molecule of  claim 10 , wherein each Linker comprises an amino acid sequence independently selected from SEQ ID NO: 12 and SEQ ID NO: 16. 
     
     
         12 . A tetravalent GITR-targeting molecule comprising two copies of the polypeptide of  claim 3 . 
     
     
         13 . A pharmaceutical composition comprising the tetravalent GITR-targeting molecule of  claim 4 . 
     
     
         14 . A method of treating cancer comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of  claim 13 . 
     
     
         15 . The method of  claim 14 , wherein the cancer is selected from the group consisting of bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, and virus-related cancer. 
     
     
         16 . The method of  claim 15 , wherein the cancer is selected from the group consisting of lung cancer, head and neck cancer, pancreatic cancer, kidney cancer, ovarian cancer, neoplasm of the central nervous system, skin cancer, and thyroid cancer. 
     
     
         17 . The method of  claim 14 , further comprising administering an anti-PD1 or anti-PDL1 antibody. 
     
     
         18 . The method of  claim 17 , wherein an anti-PD1 antibody is administered. 
     
     
         19 . The method of  claim 18 , wherein the anti-PD1 antibody is BMS-936558. 
     
     
         20 . A pharmaceutical composition comprising the tetravalent GITR-targeting molecule of  claim 12 . 
     
     
         21 . A method of treating cancer comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of  claim 20 . 
     
     
         22 . The method of  claim 21 , wherein the cancer is selected from the group consisting of bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, and virus-related cancer. 
     
     
         23 . The method of  claim 22 , wherein the cancer is selected from the group consisting of lung cancer, head and neck cancer, pancreatic cancer, kidney cancer, ovarian cancer, neoplasm of the central nervous system, skin cancer, and thyroid cancer. 
     
     
         24 . The method of  claim 23 , further comprising administering an anti-PD1 or anti-PDL1 antibody. 
     
     
         25 . The method of  claim 24 , wherein an anti-PD1 antibody is administered. 
     
     
         26 . The method of  claim 25 , wherein the anti-PD1 antibody is BMS-936558. 
     
     
         27 . The polypeptide of  claim 1 , wherein the GITR-BD comprises an amino acid sequence selected from SEQ ID NOs: 50-59 and 61-62. 
     
     
         28 . The polypeptide of  claim 4 , wherein each GITR-BD comprises an amino acid sequence selected from SEQ ID NOs: 50-59 and 61-62.