Targeted protein degradation to attenuate adoptive t-cell therapy associated adverse inflammatory responses

Abstract

This invention is in the area of compositions and methods for regulating chimeric antigen receptor immune effector cell, for example T-cell (CAR-T), therapy to modulate associated adverse inflammatory responses, for example, cytokine release syndrome and tumor lysis syndrome, using targeted protein degradation.

1 . A method of reducing an adverse immune response in a subject caused by an activated immune effector cell that expresses a chimeric antigen receptor polypeptide comprising:
 administering to the subject experiencing an adverse immune response an effective amount of a heterobifunctional compound;   wherein the subject has previously been administered an immune effector cell capable of expressing a chimeric antigen receptor polypeptide;   wherein the chimeric antigen receptor polypeptide comprises:   i) an extracellular ligand binding protein;   ii) a transmembrane protein;   iii) a cytoplasmic protein comprising at least one intracellular signaling protein; and,   iv) a heterobifunctional compound targeting protein capable of being bound by a heterobifunctional compound;   wherein the administered heterobifunctional compound binds to i) the chimeric receptor antigen polypeptide through the heterobifunctional compound targeting protein and ii) a ubiquitin ligase in a manner that brings the chimeric antigen receptor polypeptide into proximity of the ubiquitin ligase; and   wherein the chimeric antigen receptor polypeptide, when bound by the heterobifunctional compound, is ubiquitinated and then degraded by a proteasome.   
     
     
         2 . The method of  claim 1 , wherein the immune effector cell is an autologous human cell. 
     
     
         3 . The method of  claim 1 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence from a non-endogenous peptide. 
     
     
         4 . The method of  claim 1 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence selected from SEQ ID NO: 1-9 and 24-58. 
     
     
         5 . The method of  claim 4 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 1. 
     
     
         6 . The method of  claim 5 , wherein the heterobifunctional compound targeting protein is capable of being bound by a heterobifunctional compound selected from dFKBP1 to dFKBP13. 
     
     
         7 . The method of  claim 4 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 2. 
     
     
         8 . The method of  claim 4 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 3. 
     
     
         9 . The method of  claim 8 , wherein the heterobifunctional compound targeting protein is capable of being bound by a heterobifunctional compound selected from dBET1 to dBET18. 
     
     
         10 . The method of  claim 4 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 9. 
     
     
         11 . The method of  claim 10 , wherein the heterobifunctional compound targeting protein is capable of being bound by a heterobifunctional compound selected from dHalo1 to dHalo2. 
     
     
         12 . The method of  claim 4 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 45. 
     
     
         13 .- 30 . (canceled) 
     
     
         31 . The method of  claim 1 , wherein the extracellular ligand binding protein binds CD19. 
     
     
         32 . The method of  claim 1 , wherein the transmembrane protein comprises the transmembrane region of CD28. 
     
     
         33 . The method of  claim 1 , wherein the at least one intracellular signaling protein is derived from CD3 zeta. 
     
     
         34 . The method of  claim 1 , wherein the at least one intracellular signaling protein further comprises a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83. 
     
     
         35 . The method of  claim 1 , wherein the heterobifunctional compound targeting protein comprises an amino acid sequence of SEQ ID NO: 2 and the heterobifunctional compound targeting protein is capable of being bound by a heterobifunctional compound selected from dFKBP6 to dFKBP13.