Axl tyrosine kinase inhibitors and methods of making and using the same

Abstract

Disclosed are novel inhibitors of the Axl receptor tyrosine kinase (RTK) and methods of using such inhibitors in a variety of therapeutic approaches in the areas of cancer therapy and anti-thrombosis (anti-clotting) therapy.

What is claimed is: 
     
         1 . An Axl fusion protein comprising:
 a) a first protein comprising, consisting essentially of, or consisting of, at least a portion of the extracellular domain of an Axl receptor tyrosine kinase (Axl RTK) that binds to an Axl ligand; and   b) a second protein that is a heterologous fusion protein, wherein the second protein is fused to the first protein.   
     
     
         2 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of the Gas6 major binding site of Axl. 
     
     
         3 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of the Gas6 major binding site and the Gas6 minor binding site of Axl. 
     
     
         4 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of the Ig1 domain of Axl. 
     
     
         5 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of the Ig1 domain and the Ig2 domain of Axl. 
     
     
         6 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of a portion of the extracellular domain of Axl RTK in which at least one of the FBNIII motifs in the first protein is deleted or mutated of Axl. 
     
     
         7 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of a portion of the extracellular domain of Axl RTK in which both of the FBNIII motifs is deleted or mutated of Axl. 
     
     
         8 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of, the entire Axl RTK extracellular domain of Axl. 
     
     
         9 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of positions 1-445 of Axl RTK, with respect to SEQ ID NO:2. 
     
     
         10 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of positions 1-325 of Axl RTK, with respect to SEQ ID NO:2. 
     
     
         11 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of positions 1-225 of Axl RTK, with respect to SEQ ID NO:2. 
     
     
         12 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of at least: positions 10-222 of Axl RTK, positions 20-222 of Axl RTK, positions 30-222 of Axl RTK, positions 40-222 of Axl RTK, positions 50-222 of Axl RTK, or positions 60-222 of Axl RTK, with respect to SEQ ID NO:2. 
     
     
         13 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of at least: positions 10-225 of Axl RTK, positions 20-225 of Axl RTK, positions 30-225 of Axl RTK, positions 40-225 of Axl RTK, positions 50-225 of Axl RTK, or positions 60-225 of Axl RTK, with respect to SEQ ID NO:2. 
     
     
         14 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of: at least positions 63-225 of SEQ ID NO:2. 
     
     
         15 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of at least: positions 1-137 of Axl RTK, positions 10-137 of Axl RTK, positions 20-137 of Axl RTK, positions 30-137 of Axl RTK, positions 40-137 of Axl RTK, positions 50-137 of Axl RTK, or positions 60-137 or Axl RTK, with respect to SEQ ID NO:2. 
     
     
         16 . The Axl fusion protein of  claim 1 , wherein the first protein comprises, consists essentially of, or consists of at least positions 63 to 218 of SEQ ID NO:2. 
     
     
         17 . The Axl fusion protein of  claim 1 , wherein the first protein comprises at least positions 63-99, 136, 138, and 211-218 of SEQ ID NO:2, arranged in a conformation that retains the tertiary structure of these positions with respect to the full-length extracellular domain of Axl RTK (positions 1-445 of SEQ ID NO:2). 
     
     
         18 . The Axl fusion protein of  claim 1 , wherein the Axl RTK comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:2 or SEQ ID NO:4. 
     
     
         19 . The Axl fusion protein of  claim 1 , wherein the Axl RTK comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:4. 
     
     
         20 . The Axl fusion protein of  claim 1 , wherein the Axl RTK comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:4. 
     
     
         21 . The Axl fusion protein of  claim 1 , wherein the Axl RTK comprises an amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4. 
     
     
         22 . The Axl fusion protein of  claim 1 , wherein the fusion protein is produced as a dimer of Axl proteins. 
     
     
         23 . The Axl fusion protein of  claim 1 , wherein the heterologous fusion protein is an immunoglobulin Fc domain. 
     
     
         24 . The Axl fusion protein of  claim 23 , wherein the immunoglobulin Fc domain consists essentially of or consists of a heavy chain hinge region, a CH 2  domain and a CH 3  domain. 
     
     
         25 . The Axl fusion protein of  claim 23 , wherein the immunoglobulin Fc domain is from an IgG immunoglobulin protein. 
     
     
         26 . The Axl fusion protein of  claim 23 , wherein the immunoglobulin Fc domain is from an IgG1 immunoglobulin protein. 
     
     
         27 . The Axl fusion protein of  claim 23 , wherein the immunoglobulin Fc domain is from a human immunoglobulin. 
     
     
         28 - 37 . (canceled) 
     
     
         38 . An Axl fusion protein comprising:
 a) a first protein comprising, consisting essentially of, or consisting of, at least a portion of the extracellular domain of an Axl receptor tyrosine kinase (Axl RTK) that binds to a receptor selected from the group consisting of Axl, Tyro and Mer and inhibits the binding of a ligand to said receptor or inhibits dimerization, trimerization or formation of any receptor-protein complex of said receptor; and   b) a second protein that is a heterologous fusion protein, wherein the second protein is fused to the first protein.   
     
     
         39 . The Axl fusion protein of  claim 38 , wherein the first protein consists of the FNIII domains of Axl. 
     
     
         40 - 62 . (canceled)