Anti-CD19 humanized antibody and immune effector cell targeting cd 19
Disclosed are an anti-CD19 humanized antibody prepared from a murine monoclonal antibody, a chimeric antigen receptor containing the humanized antibody, and an immune cell expressing the humanized antibody. Not only does the humanized antibody of the present invention not produce an anti-antibody response (AAR) and a human anti-mouse antibody response (HAMA), but same also has better affinity than a murine antibody, and has excellent activity and safety, thereby providing a new means for treating CD19-expressing tumors.
1. An antibody that binds CD19, wherein the antibody comprises a light chain variable region (VL) comprising light chain complementarity determining region (LCDR) 1, 2, and 3 and comprises a heavy chain variable region (VH) comprising heavy chain complementarity determining region (HCDR) 1, 2, and 3; wherein the antibody is selected from the group consisting of:
(a) an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11;
(b) an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region of SEQ ID NO: 5;
(c) an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region of SEQ ID NO: 3;
(d) an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11;
(e) an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region of SEQ ID NO: 5;
(f) an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region of SEQ ID NO: 3;
(g) an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11;
(h) an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region of SEQ ID NO: 5; and
(i) an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region of SEQ ID NO: 3.
2. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11.
3. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region of SEQ ID NO: 5.
4. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 1 and a heavy chain variable region of SEQ ID NO: 3.
5. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11.
6. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region of SEQ ID NO: 5.
7. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region of SEQ ID NO: 3.
8. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11.
9. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region of SEQ ID NO: 5.
10. The antibody of claim 1 , wherein the antibody comprises an antibody having a light chain variable region of SEQ ID NO: 7 and a heavy chain variable region of SEQ ID NO: 3.
11. The antibody of claim 1 , wherein the antibody is selected from the group consisting of a Fab, Fab′, Fab′-SH, Fd, F(ab′)2, scFv, bispecific single chain Fv dimer, diabodies, tribodies, and scFvs genetically fused to the same or different antibodies.
12. A CD19 specific chimeric antigen receptor (CAR) comprising an extracellular domain, a hinge domain, a transmembrane domain, and an intracellular signal domain sequentially linked, wherein the extracellular domain comprises an anti-CD19 antibody of claim 1 .
13. The chimeric antigen receptor of claim 12 , wherein the intracellular signal domain further comprises one or more co-stimulatory signal domains.
14. The chimeric antigen receptor of claim 13 , wherein the transmembrane domain is a transmembrane domain of CD8α or CD28; or wherein the co-stimulatory signal domain is the intracellular signal domain of CD137 or CD28.
15. The chimeric antigen receptor of claim 12 , wherein the extracellular domain in a single chain antibody comprising a light chain variable region that has LCDR1 as shown in SEQ ID NO: 17, LCDR2 as shown in SEQ ID NO: 13 and LCDR3 as shown in SEQ ID NO: 14 and a heavy chain variable region that has HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 16 and HCDR3 as shown in SEQ ID NO: 11.
16. A genetically modified immune cell which expresses the chimeric antigen receptor of claim 12 .
17. The genetically modified immune cell of claim 16 , wherein the genetically modified immune cell further expresses a sequence that is a cytokine selected from IL-12, IL-15, IL-21 or type I interferon or a sequence that is a safety switch.
18. A genetically modified T cell which expresses the chimeric antigen receptor of claim 15 .
19. A multifunctional immunoconjugate, comprising an antibody of claim 1 and a functional molecule linked thereto, wherein the functional molecule is selected from the group consisting of a molecule that targets tumor surface markers other than CD19, a molecule that inhibits tumors, a molecule that targets a surface marker on an immune cell, or a detectable label; wherein the multifunctional immunoconjugate optionally comprises a linker peptide between the antibody and functional molecule.
20. The multifunctional immunoconjugate of claim 19 , wherein the molecule that inhibits tumors is an antitumor cytokine or an antitumor toxin.
21. The multifunctional immunoconjugate of claim 19 , wherein the molecule that targets a surface marker of an immune cell is an anti-CD3 antibody.
22. A pharmaceutical composition comprising an antibody of claim 1 .
23. A pharmaceutical composition comprising genetically modified T cells of claim 18 .
24. A pharmaceutical composition comprising a multifunctional immunoconjugate of claim 19 .
25. A CD19 specific chimeric antigen receptor, wherein the chimeric antigen receptor is selected from the group consisting of:
chimeric antigen receptor huHD37-28Z, which has an extracellular domain as shown in SEQ ID NO: 21, a hinge domain as shown in SEQ ID NO: 25, a transmembrane domain as shown in SEQ ID NO: 27, a co-stimulatory signal domain as shown in SEQ ID NO: 29, and a primary signal domain as shown in SEQ ID NO: 31;
chimeric antigen receptor huHD37-BBZ, which has an extracellular domain as shown in SEQ ID NO: 21, a hinge domain as shown in SEQ ID NO: 25, a transmembrane domain as shown in SEQ ID NO: 33, a co-stimulatory signal domain as shown in SEQ ID NO:35, and a primary signal domain as shown in SEQ ID NO: 31; and
chimeric antigen receptor huHD37-28BBZ, which has an extracellular domain as shown in SEQ ID NO: 21, a hinge domain as shown in SEQ ID NO: 25, a transmembrane domain as shown in SEQ ID NO: 27, costimulatory signal domains as shown in SEQ ID NO: 29 and SEQ ID NO: 35, and a primary signal domain as shown in SEQ ID NO: 31.
26. A genetically modified T cell which expresses a CD19 specific chimeric antigen receptor of claim 25 .
27. A pharmaceutical composition comprising genetically modified T cells of claim 26 .
28. A method of killing a CD19-expressing tumor cell comprising contacting the tumor cell with genetically modified T cells of claim 26 .