CHIMERIC ANTIGEN RECEPTORS (CARs), COMPOSITIONS AND METHODS THEREOF

Abstract

The present disclosure relates to compositions and methods relating to chimeric antigen receptor (CAR) polypeptides and methods relating thereto. In one embodiment, the present disclosure relates to engineered cells having chimeric antigen receptor polypeptides directed to at least two targets. In another embodiment, the present disclosure relates to engineered cells having chimeric antigen receptor polypeptides and an enhancer moiety.

1 .- 105 . (canceled) 
     
     
         106 . An engineered cell comprising:
 (i) a first chimeric antigen receptor polypeptide comprising a first antigen recognition domain, a first signal peptide, a first hinge region, a first transmembrane domain, a first co-stimulatory domain, and a first signaling domain;   (ii) a second chimeric antigen receptor polypeptide comprising a second antigen recognition domain, a second signal peptide, a second hinge region, a second transmembrane domain, a second co-stimulatory domain, and a second signaling domain; and   
       wherein 
       the first antigen recognition domain is selective for one of interleukin 6 receptor, NY-ESO-1, alpha fetoprotein (AFP), glypican-3 (GPC3), BCMA, BAFF, BAFF-R, BCMA, TACI, LeY, CD4, CD25, CD38, CD5, CD13, CD14, CD15 CD19, CD20, CD22, CD33, CD41, CD61, CD64, CD68, CD117, CD123, CD138, CD267, CD269, CD38, Flt3 receptor, APRIL, and CS1; and 
       the second antigen recognition domain is selective for one of interleukin 6 receptor, NY-ESO-1, alpha fetoprotein (AFP), glypican-3 (GPC3), BCMA, BAFF, BAFF-R, BCMA, TACI, LeY, CD4, CD25, CD38, CD5, CD13, CD14, CD15 CD19, CD20, CD22, CD33, CD41, CD61, CD64, CD68, CD117, CD123, CD138, CD267, CD269, CD38, Flt3 receptor, APRIL, and CS1; 
       the first antigen recognition domain and the second antigen recognition domain are different; 
       the first chimeric antigen receptor polypeptide and the second chimeric antigen receptor polypeptide each consist of a single antigen recognition domain; and 
       the first antigen recognition domain and second antigen recognition domain are expressed on the surface of the engineered cell. 
     
     
         107 . The engineered cell according to  claim 106 , wherein the first chimeric antigen receptor polypeptide and the second chimeric engineered polypeptide are on a single polypeptide molecule, and wherein an amino acid sequence comprising a high efficiency cleavage site is disposed between the first chimeric antigen receptor polypeptide and the second chimeric antigen receptor polypeptide, and wherein high efficiency cleavage site is defined as a polypeptide sequence that results in greater than 80% self cleavage. 
     
     
         108 . The engineered cell according to  claim 107 , wherein the high efficiency cleavage site is selected from the group consisting of P2A, T2A, E2A, and F2A. 
     
     
         109 . The engineered cell according to  claim 106  wherein the first co-stimulatory domain and the second co-stimulatory domain are different. 
     
     
         110 . The engineered cell according to  claim 106 , wherein the first co-stimulatory domain comprises CD28, and the second co-stimulatory domain comprises 4-1BB. 
     
     
         111 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for TACI or CD269; and the second antigen recognition domain is selective for one of CD19, CD38, CD138, CD138, and CS1. 
     
     
         112 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for CD19; and the second antigen recognition domain is selective for one of CD20, CD22, CD33, CD123, TACI, CD269, CD38, and CS1. 
     
     
         113 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for CD33; and the second antigen recognition domain is selective for LeY or CD123. 
     
     
         114 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for BCMA; and the second antigen recognition domain is selective for one of CS1, CD19, CD20, CD22, CD38, CD138, and CS1. 
     
     
         115 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for CD19; and the second antigen recognition domain is selective for BCMA. 
     
     
         116 . The engineered cell according to  claim 106 , wherein the engineered cell is a T cell, NK cell, NK T cell, or NK-92 cell. 
     
     
         117 . The engineered cell according to  claim 106 , wherein the engineered cell further comprises a heterologously expressed enhancer. 
     
     
         118 . The engineered cell according to  claim 117 , wherein said enhancer is selected from the group consisting of PD-1, PD-L1, CSFIR, CTAL-4, TIM-3, TGFR beta, IL-2, IL-15/IL-15 sushi, IL-6, IL-7, IL-12, IL-15, IL-17, IL-18 IL-21, functional fragments thereof, and combinations thereof. 
     
     
         119 . The engineered cell according to  claim 106 , wherein the first antigen recognition domain is selective for one of CD19, CD33, and CD269; and the second antigen recognition domain is selective for CD123 or CS1. 
     
     
         120 . The engineered cell according to  claim 106 , wherein TAC1 antigen recognition domain comprises the APRIL ligand or the BAFF ligand or a portion thereof. 
     
     
         121 . The engineered cell according to  claim 106 , wherein the BCMA antigen recognition domain comprises APRIL ligand or BAFF ligand or a portion thereof. 
     
     
         122 . The engineered cell according to  claim 106 , wherein the BAFF-R antigen recognition domain comprises the BAFF ligand or a portion thereof. 
     
     
         123 . The engineered cell according to  claim 106 , wherein the first co-stimulatory domain and the second co-stimulatory domain comprise 4-1BB co-stimulatory domain. 
     
     
         124 . A method of treating a cell proliferative disease in a patient, said method comprising:
 administering to the patient an engineered cell that expresses:   (i) a first chimeric antigen receptor polypeptide comprising a first antigen recognition domain, a first signal peptide, a first hinge region, a first transmembrane domain, a first co-stimulatory domain, and a first signaling domain;   (ii) a second chimeric antigen receptor polypeptide comprising a second antigen recognition domain, a second signal peptide, a second hinge region, a second transmembrane domain, a second co-stimulatory domain, and a second signaling domain; and   reducing the tumor burden of cell proliferative disease cells;   
       wherein 
       the first antigen recognition domain is selective for one of interleukin 6 receptor, NY-ESO-1, alpha fetoprotein (AFP), glypican-3 (GPC3), BCMA, BAFF, BAFF-R, BCMA, TACI, LeY, CD4, CD25, CD38, CD5, CD13, CD14, CD15 CD19, CD20, CD22, CD33, CD41, CD61, CD64, CD68, CD117, CD123, CD138, CD267, CD269, CD38, Flt3 receptor, APRIL, and CS1; and 
       the second antigen recognition domain is selective for one of interleukin 6 receptor, NY-ESO-1, alpha fetoprotein (AFP), glypican-3 (GPC3), BCMA, BAFF, BAFF-R, BCMA, TACI, LeY, CD4, CD25, CD38, CD5, CD13, CD14, CD15 CD19, CD20, CD22, CD33, CD41, CD61, CD64, CD68, CD117, CD123, CD138, CD267, CD269, CD38, Flt3 receptor, APRIL, and CS1; 
       the first antigen recognition domain and the second antigen recognition domain are different; 
       the first chimeric antigen receptor polypeptide and the second chimeric antigen receptor polypeptide each consist of a single antigen recognition domain; and 
       the first antigen recognition domain and second antigen recognition domain are expressed on the surface of the engineered cell. 
     
     
         125 . The method according to  claim 124 , wherein the engineered cell comprises T cells, NK cells, NK T cells, or NK-92 cells. 
     
     
         126 . The method according to  claim 124 , wherein the cell proliferative disease is selected from the group consisting of: B-cell lymphoma, T-cell lymphoma, multiple myeloma, chronic myeloid leukemia, B-cell acute lymphoblastic leukemia (B-ALL), and plasma cell neoplasms.