Ciliary Neurotrophic Factor Receptor Ligand-Binding Agents and Methods of Using the Same

Abstract

Provided are agents that specifically bind a ligand of ciliary neurotrophic factor receptor (CNTFR). In certain aspects, an agent of the present disclosure is a soluble CNTFR polypeptide. The soluble CNTFR polypeptide may have an altered (e.g., reduced) binding affinity for one or more ligand-CNTFR complex subunits, an altered (e.g., increased) binding affinity for one or more CNTFR ligands, or any combination thereof. Compositions that include the agents of the present disclosure are also provided, as are methods of using the agents (e.g., for treating a cell proliferative disorder) and methods of identifying an individual as having a cell proliferative disorder associated with CNTFR signaling.

1 - 89 . (canceled) 
     
     
         90 . A method of treating a disorder associated with CNTFR signaling in an individual in need thereof, the method comprising:
 administering to the individual a therapeutically effective amount of a soluble ciliary neurotrophic factor receptor (CNTFR) polypeptide comprising a CNTFR extracellular domain that specifically binds cardiotrophin-like cytokine factor 1 (CLCF1), wherein the extracellular domain comprises:
 one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for leukemia inhibitory factor receptor (LIFR) relative to a wild-type CNTFR polypeptide, wherein the one or more mutations comprise a mutation at amino acid position 177, 178, or both, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1; and/or 
 one or more mutations that increase binding affinity of the soluble CNTFR polypeptide for CLCF1 relative to a wild-type CNTFR polypeptide, wherein the one or more mutations comprise a mutation at amino acid position 110, 174, 237, 287, or any combination thereof, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1. 
   
     
     
         91 . The method according to  claim 90 , wherein the one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for LIFR comprise Y177H, K178N, or both. 
     
     
         92 . The method according to  claim 90 , wherein the one or more mutations that increase binding affinity of the soluble CNTFR polypeptide for CLCF1 comprise R110Q, T174P, S237F, I287F, or any combination thereof. 
     
     
         93 . The method according to  claim 90 , wherein the soluble CNTFR polypeptide comprises a mutation at each of amino acid positions 110, 174, 177, 178, 237, and 287. 
     
     
         94 . The method according to  claim 93 , wherein the soluble CNTFR polypeptide comprises each of the following mutations: R110Q, T174P, Y177H, K178N, S237F, and I287F. 
     
     
         95 . The method according to  claim 90 , wherein the soluble CNTFR polypeptide comprises one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for glycoprotein 130 (gp130) relative to a wild-type CNTFR polypeptide. 
     
     
         96 . The method according to  claim 95 , wherein the one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for gp130 comprise a mutation at amino acid position 268, 269, or both, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1. 
     
     
         97 . The method according to  claim 96 , wherein the one or more mutations that reduce binding affinity for gp130 comprise T268A, D269A, or both. 
     
     
         98 . The method according to  claim 90 , wherein the individual in need thereof has a cell proliferative disorder associated with CNTFR signaling, and the administering is effective in treating the cell proliferative disorder. 
     
     
         99 . The method according to  claim 98 , wherein the cell proliferative disorder is cancer. 
     
     
         100 . The method according to  claim 99 , wherein the cancer is lung cancer. 
     
     
         101 . The method according to  claim 100 , wherein the lung cancer is non-small cell lung cancer (NSCLC). 
     
     
         102 . A pharmaceutical composition comprising:
 a soluble CNTFR polypeptide comprising a CNTFR extracellular domain that specifically binds CLCF1, wherein the extracellular domain comprises:
 one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for LIFR relative to a wild-type CNTFR polypeptide, wherein the one or more mutations comprise a mutation at amino acid position 177, 178, or both, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1; and/or 
 one or more mutations that increase binding affinity of the soluble CNTFR polypeptide for CLCF1 relative to a wild-type CNTFR polypeptide, wherein the one or more mutations comprise a mutation at amino acid position 110, 174, 237, 287, or any combination thereof, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1; and 
   a pharmaceutically acceptable carrier.   
     
     
         103 . The pharmaceutical composition of  claim 102 , wherein the one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for LIFR comprise Y177H, K178N, or both. 
     
     
         104 . The pharmaceutical composition of  claim 102 , wherein the one or more mutations that increase binding affinity of the soluble CNTFR polypeptide for CLCF1 comprise R110Q, T174P, S237F, 1287F, or any combination thereof. 
     
     
         105 . The pharmaceutical composition of  claim 102 , wherein the soluble CNTFR polypeptide comprises a mutation at each of amino acid positions 110, 174, 177, 178, 237, and 287. 
     
     
         106 . The pharmaceutical composition of  claim 105 , wherein the soluble CNTFR polypeptide comprises each of the following mutations: R110Q, T174P, Y177H, K178N, S237F, and 1287F. 
     
     
         107 . The pharmaceutical composition of  claim 102 , wherein the soluble CNTFR polypeptide comprises one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for glycoprotein 130 (gp130) relative to a wild-type CNTFR polypeptide. 
     
     
         108 . The pharmaceutical composition of  claim 102 , wherein the soluble CNTFR polypeptide comprises one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for glycoprotein 130 (gp130) relative to a wild-type CNTFR polypeptide. 
     
     
         109 . The pharmaceutical composition of  claim 108 , wherein the one or more mutations that reduce binding affinity of the soluble CNTFR polypeptide for gp130 comprise a mutation at amino acid position 268, 269, or both, relative to a CNTFR polypeptide having the amino acid sequence set forth in SEQ ID NO:1.