Synergistic tumor treatment with il-2 and integrin-binding-fc fusion protein

Abstract

The present invention provides a method of treating cancer with a combination of IL-2 and an integrin-binding-Fc fusion protein. The methods of the invention can be applied to a broad range of cancer types.

1 . A method for treating cancer in a subject comprising administering to the subject an effective amount of interleukin (IL)-2 and an integrin-binding-Fc fusion protein, wherein the integrin-binding-Fc fusion protein comprises (i) an integrin-binding polypeptide comprising an integrin-binding loop and a knottin polypeptide scaffold; and (ii) an immunoglobulin Fc domain, wherein the integrin-binding polypeptide is operably linked to the Fc domain. 
     
     
         2 . The method of  claim 1 , wherein IL-2 is an extended pharmacokinetic (PK) IL-2. 
     
     
         3 . The method of  claim 1 , wherein the extended-PK IL-2 comprises a fusion protein. 
     
     
         4 . The method of  claim 3  wherein the fusion protein comprises an IL-2 moiety and a moiety selected from the group consisting of an immunoglobulin fragment, human serum albumin, and Fn3. 
     
     
         5 . The method of  claim 4 , wherein the fusion protein comprises an IL-2 moiety operably linked to an immunoglobulin Fc domain. 
     
     
         6 . The method of  claim 4 , wherein the fusion protein comprises an IL-2 moiety operably linked to human serum albumin. 
     
     
         7 . The method of  claim 1 , wherein the extended-PK IL-2 comprises an IL-2 moiety conjugated to a non-protein polymer. 
     
     
         8 . The method of  claim 7 , wherein the non-protein polymer is a polyethylene glycol. 
     
     
         9 . The method of  claim 1 , wherein the integrin-binding polypeptide binds to a tumor-associated integrin selected from the group consisting of α v β 3 , α v β 5 , and α 5 β 1 , or combination thereof. 
     
     
         10 . The method of  claim 1 , wherein the integrin-binding polypeptide binds to α v β 3 , α v β 5 , and α 5 β 1 . 
     
     
         11 . The method of  claim 1 , wherein the knottin polypeptide scaffold comprises at least three cysteine disulfide linkages or crosslinked cysteine residues, and wherein the integrin-binding loop is adjacent to cysteine residues of the knottin polypeptide scaffold. 
     
     
         12 . The method of  claim 11 , wherein the integrin-binding loop comprises an RGD peptide sequence. 
     
     
         13 . The method of  claim 11 , wherein the knottin polypeptide scaffold is derived from a knottin protein selected from the group consisting of EETI-II, AgRP, and agatoxin. 
     
     
         14 . The method of  claim 13 , wherein the knottin protein is EETI-II. 
     
     
         15 . The method of  claim 1 , wherein the integrin-binding loop comprises an RGD peptide sequence and the knottin polypeptide scaffold is derived from EETI-II. 
     
     
         16 . The method of  claim 1 , wherein the knottin polypeptide scaffold is derived from EETI-II and the integrin-binding loop comprises the sequence X 1 X 2 X 3 RGDX 7 X 8 X 9 X 10 X 11 , wherein each X represents any amino acid, wherein the loop is inserted between 2 cysteine residues in the EETI-II sequence and replaces the native EETI-II sequence. 
     
     
         17 . The method of  claim 16 , wherein the integrin-binding loop is inserted after the first cysteine in the native EETI-II sequence. 
     
     
         18 .- 34 . (canceled) 
     
     
         35 . A method for inhibiting growth and/or proliferation of tumor cells in a subject comprising administering to the subject an effective amount of an integrin-binding-Fc fusion protein, wherein the integrin-binding-Fc fusion protein comprises (i) an integrin-binding polypeptide comprising an integrin-binding loop and a knottin polypeptide scaffold; and (ii) an immunoglobulin Fc domain, wherein the integrin-binding polypeptide is operably linked to the Fc domain, and an immune checkpoint blocker. 
     
     
         36 .- 38 . (canceled) 
     
     
         39 . A method for inhibiting growth and/or proliferation of tumor cells in a subject comprising administering to the subject an effective amount of an integrin-binding-Fc fusion protein, wherein the integrin-binding-Fc fusion protein comprises (i) an integrin-binding polypeptide comprising an integrin-binding loop and a knottin polypeptide scaffold; and (ii) an immunoglobulin Fc domain, wherein the integrin-binding polypeptide is operably linked to the Fc domain, and an antagonist of VEGF. 
     
     
         40 . The method of  claim 39 , wherein the antagonist of VEGF is an antibody or antibody fragment thereof that binds VEGF, an antibody or antibody fragment thereof that binds VEGF receptor, a small molecule inhibitor of the VEGF receptor tyrosine kinase, a dominant negative VEGF, or a VEGF receptor.