Anti-cd276 polypeptides, proteins, and chimeric antigen receptors

Abstract

Polypeptides and proteins that specifically bind to and immunologically recognize CD276 are disclosed. Chimeric antigen receptors (CARs), anti-CD276 binding moieties, nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the polypeptides and proteins are also disclosed. Methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal are also disclosed.

1 - 29 . (canceled) 
     
     
         30 . A polypeptide comprising
 (a) a heavy and light chain comprising (i) SEQ ID NOs: 1-6;
 a heavy and light chain comprising (ii) SEQ ID NOs: 11-16; or 
 a heavy and light chain comprising (iii) SEQ ID NOs: 20-25; 
 wherein the heavy and light chains of (i), (ii), or (iii) bind to CD276 and are linked to one or more of the immunoglobulin constant domains CH2 and/or CH3, and 
   (b) a leader amino acid sequence.   
     
     
         31 . The polypeptide according to  claim 30 , wherein the leader amino acid sequence comprises SEQ ID NO: 60, SEQ ID NO: 61 or SEQ ID NO: 62. 
     
     
         32 . The polypeptide according to  claim 30 , wherein the one or more immunoglobulin constant domains comprise SEQ ID NO: 71. 
     
     
         33 . The polypeptide according to  claim 30 , wherein the heavy and light chains of (i), (ii), or (iii) and the one or more immunoglobulin constant domains comprise SEQ ID NO: 80, SEQ ID NO: 82, or SEQ ID NO: 84. 
     
     
         34 . A protein comprising:
 (a) a first polypeptide chain comprising (i) a heavy chain comprising SEQ ID NOs: 1-3,
 (ii) a heavy chain comprising SEQ ID NOs: 11-13, or 
 (iii) a heavy chain comprising SEQ ID NOs: 20-22; and 
   (b) a second polypeptide chain comprising
 (iv) a light chain comprising SEQ ID NOs: 4-6, 
 (v) a light chain comprising SEQ ID NOs: 14-16, or 
 (vi) a light chain comprising SEQ ID NOs: 23-25, 
 wherein the first polypeptide of (i), (ii), or (iii) and the second polypeptide of (iv), (v), or (vi) bind to CD276, and are linked to one or more of the immunoglobulin constant domains CH2 and/or CH3, and 
   (c) a leader amino acid sequence.   
     
     
         35 . The protein according to  claim 34 , wherein the leader amino acid sequence comprises SEQ ID NO: 60, SEQ ID NO: 61 or SEQ ID NO: 62. 
     
     
         36 . The protein according to  claim 34 , wherein the one or more immunoglobulin constant domains comprise SEQ ID NO: 71. 
     
     
         37 . The protein according to  claim 34 , wherein the first polypeptide chain, second polypeptide chain, and one or more immunoglobulin constant domains comprise SEQ ID NO: 80, SEQ ID NO: 82, or SEQ ID NO: 84. 
     
     
         38 . The polypeptide according to  claim 30 , further comprising a linker amino acid sequence. 
     
     
         39 . The polypeptide according to  claim 38 , wherein the linker comprises SEQ ID NO: 9 or SEQ ID NO: 115. 
     
     
         40 . The polypeptide according to  claim 30 , comprising SEQ ID NO: 10, SEQ ID NO: 19, or SEQ ID NO: 28. 
     
     
         41 . A chimeric antigen receptor (CAR) comprising
 (a) an antigen binding domain formed by a heavy and light chain comprising (i) SEQ ID NOs: 1-6, a heavy and light chain comprising (ii) SEQ ID NOs: 11-16, or a heavy and light chain comprising (iii) SEQ ID NOs: 20-25, wherein the heavy and light chains of (i), (ii), or (iii) bind to CD276;   (b) an immunoglobulin constant domain;   (c) a leader amino acid sequence;   (d) a transmembrane domain, and   (e) an intracellular T cell signaling domain.   
     
     
         42 . The CAR according to  claim 41 , wherein the leader amino acid sequence comprises SEQ ID NO: 60, SEQ ID NO: 61 or SEQ ID NO: 62. 
     
     
         43 . The CAR according to  claim 41 , wherein the immunoglobulin constant domain comprises one or more of CH2 and/or CH3 and is effective to extend the antigen binding domain away from a membrane of a CAR expressing cell. 
     
     
         44 . The CAR according to  claim 41 , wherein the immunoglobulin constant domain comprises SEQ ID NO: 71. 
     
     
         45 . The CAR according to  claim 41 , wherein the transmembrane domain comprises CD8 and CD28. 
     
     
         46 . The CAR according to  claim 41 , wherein the transmembrane domain comprises any one or more of a CD8 amino acid sequence comprising SEQ ID NO: 29, a CD28 amino acid sequence comprising SEQ ID NO: 30, and a CD8 amino acid sequence comprising SEQ ID NO: 31. 
     
     
         47 . The CAR according to  claim 41 , wherein the intracellular T cell signaling domain comprises one or more of i) CD28, ii) CD137, and iii) CD3 zeta. 
     
     
         48 . The CAR according to  claim 41 , wherein the intracellular T cell signaling domain comprises a CD28 amino acid sequence comprising SEQ ID NO: 32 and/or SEQ ID NO: 35. 
     
     
         49 . The CAR according to  claim 41 , wherein the intracellular T cell signaling domain comprises a CD137 amino acid sequence comprising SEQ ID NO: 33 and/or SEQ ID NO: 37. 
     
     
         50 . A conjugate comprising (a) the polypeptide of  claim 30  conjugated to (b) an effector molecule. 
     
     
         51 . The conjugate according to  claim 50 , wherein the effector molecule is PE4E, PE40, PE38, PE25, PE38QQR, PE38KDEL, PE-LR, or PE35. 
     
     
         52 . A nucleic acid comprising a nucleotide sequence encoding the polypeptide of  claim 30 . 
     
     
         53 . The nucleic acid according to  claim 52  comprising a nucleotide sequence comprising SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 72, SEQ ID NOs: 76-79, SEQ ID NO: 81, SEQ ID NO: 83 or SEQ ID NO: 140. 
     
     
         54 . A recombinant expression vector comprising the nucleic acid of  claim 52  or  claim 53 . 
     
     
         55 . An isolated host cell comprising the recombinant expression vector of  claim 54 . 
     
     
         56 . A population of cells comprising at least one isolated host cell of  claim 55 . 
     
     
         57 . The polypeptide of  claim 30  expressed in one or more host cells for use in treating cancer in a mammal.