Antigen-binding domain
The present disclosure relates to antigen-binding domains which bind the antigen prostate-specific membrane antigen (PSMA) and to chimeric antigen receptors (CARs) which comprise such antigen binding domains.
1 . An antigen-binding domain which binds prostate-specific membrane antigen (PSMA) and comprises either:
(a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) HCDR1, HCDR2 and HCDR3; and a light chain variable region (VL) having CDRs LCDR1, LCDR2 and LCDR3:
(i) HCDR1 comprising SEQ ID NO. 1; HCDR2 comprising SEQ ID NO. 2; HCDR3 comprising SEQ ID NO. 3; LCDR1 comprising SEQ ID NO. 4; LCDR2 comprising SEQ ID NO. 5; LCDR3 comprising SEQ ID NO. 6;
(ii) HCDR1 comprising SEQ ID NO. 9; HCDR2 comprising SEQ ID NO. 10; HCDR3 comprising SEQ ID NO. 11; LCDR1 comprising SEQ ID NO. 12; LCDR2 comprising SEQ ID NO. 13; LCDR3 comprising SEQ ID NO. 14;
(iii) HCDR1 comprising SEQ ID NO. 17; HCDR2 comprising SEQ ID NO. 18; HCDR3 comprising SEQ ID NO. 19; LCDR1 comprising SEQ ID NO. 20; LCDR2 comprising SEQ ID NO. 21; LCDR3 comprising SEQ ID NO. 22;
(iv) HCDR1 comprising SEQ ID NO. 25; HCDR2 comprising SEQ ID NO. 26; HCDR3 comprising SEQ ID NO. 27; LCDR1 comprising SEQ ID NO. 28; LCDR2 comprising SEQ ID NO. 29; LCDR3 comprising SEQ ID NO. 30;
(v) HCDR1 comprising SEQ ID NO. 33; HCDR2 comprising SEQ ID NO. 34; HCDR3 comprising SEQ ID NO. 35; LCDR1 comprising SEQ ID NO. 36; LCDR2 comprising SEQ ID NO. 37; LCDR3 comprising SEQ ID NO. 38;
(vi) HCDR1 comprising SEQ ID NO. 41; HCDR2 comprising SEQ ID NO. 42; HCDR3 comprising SEQ ID NO. 43; LCDR1 comprising SEQ ID NO. 44; LCDR2 comprising SEQ ID NO. 45; LCDR3 comprising SEQ ID NO. 46; or
(vii) HCDR1 comprising SEQ ID NO. 49; HCDR2 comprising SEQ ID NO. 50; HCDR3 comprising SEQ ID NO. 51; LCDR1 comprising SEQ ID NO. 52; LCDR2 comprising SEQ ID NO. 53; LCDR3 comprising SEQ ID NO. 54; or
(b) a domain antibody variable region having complementarity determining regions (CDRs) VHHCDR1, VHHCDR2, and VHHCDR3:
(i) VHHCDR1 comprising SEQ ID NO. 33; VHHCDR2 comprising SEQ ID NO. 34; VHHCDR3 comprising SEQ ID NO. 35;
(ii) VHHCDR1 comprising SEQ ID NO. 37; VHHCDR2 comprising SEQ ID NO. 38; VHHCDR3 comprising SEQ ID NO. 39;
(iii) VHHCDR1 comprising SEQ ID NO. 41; VHHCDR2 comprising SEQ ID NO. 42; VHHCDR3 comprising SEQ ID NO. 43;
(iv) VHHCDR1 comprising SEQ ID NO. 45; VHHCDR2 comprising SEQ ID NO. 46; VHHCDR3 comprising SEQ ID NO. 47; or
(v) VHHCDR1 comprising SEQ ID NO. 49; VHHCDR2 comprising SEQ ID NO. 50; VHHCDR3 comprising SEQ ID NO. 51.
2 . An antigen-binding domain according to claim 1 , which comprises either:
(a) a heavy chain variable region (VH); and a light chain variable region (VL):
(i) VH comprising SEQ ID NO. 7 and VL comprising SEQ ID NO. 8;
(ii) VH comprising SEQ ID NO. 15 and VL comprising SEQ ID NO. 16;
(iii) VH comprising SEQ ID NO. 23 and VL comprising SEQ ID NO. 24; or
(iv) VH comprising SEQ ID NO. 31 and VL comprising SEQ ID NO. 32; or
(b) a domain antibody variable region:
(i) SEQ ID NO. 36;
(ii) SEQ ID NO. 40;
(iii) SEQ ID NO. 44;
(iv) SEQ ID NO. 48; or
(v) SEQ ID NO. 52.
3 . A chimeric antigen receptor (CAR) comprising an antigen-binding domain according to claim 1 .
4 . A nucleic acid sequence which encodes an antigen-binding domain according to claim 1 .
5 . A nucleic acid construct which comprises a nucleic acid sequence according to claim 4 and a nucleic acid sequence which encodes a suicide gene.
6 . A vector which comprises a nucleic acid sequence according to claim 5 .
7 . A cell which expresses a CAR according to claim 3 .
8 . A method for making a cell according to claim 7 , which comprises the step of introducing a nucleic acid sequence which encodes a CAR according to claim 3 into a cell ex vivo.
9 . A pharmaceutical composition which comprises a plurality of cells according to claim 7 and a pharmaceutically acceptable carrier, diluent or excipient.
10 . A method for treating cancers that overexpress prostate specific membrane antigen which comprises the step of administering a cell according to claim 7 to a subject.