Dual function antibodies specific to glycosylated PD-L1 and methods of use thereof
Antibodies that bind specifically to glycosylated PD-L1 relative to unglycosylated PD-L1, block binding of PD-L1 to PD-1 and promote internalization and degradation of PD-L1 are provided. Antibodies that recognize specific epitopes on glycosylated PD-L1 protein and that exhibit the dual functions of both blocking the binding of PD-L1 to PD-1 and also facilitating the internalization of PD-L1 on cells are provided. In some aspects, PD-L1 polypeptides comprising glycosylated amino acid residues at amino and carboxy terminal positions of the PD-L1 extracellular domain are also provided. Methods for using such antibodies for the treatment of cancer, particularly PD-L1 positive cancer, are also provided.
1. An isolated antibody which binds to glycosylated PD-L1 comprising a V H domain comprising a CDR H1 with the amino acid sequence of SEQ ID NO: 4, a CDR H2 with the amino acid sequence of SEQ ID NO: 6, and a CDR H3 with the amino acid sequence of SEQ ID NO: 8 or a V H domain comprising a CDR H1 with the amino acid sequence of SEQ ID NO: 5, a CDR H2 with the amino acid sequence of SEQ ID NO: 7, and a CDR H3 with the amino acid sequence of SEQ ID NO: 9 and a V L domain comprising a CDR L1 with the amino acid sequence of SEQ ID NO: 12, a CDR L2 with the amino acid sequence of SEQ ID NO: 14, and a CDR L3 with the amino acid sequence of SEQ ID NO: 16.
2. The isolated antibody of claim 1 , wherein the V H domain has the amino acid sequence of SEQ ID NO: 3 and the V L has the amino acid sequence of SEQ ID NO: 11.
3. An isolated antibody which binds to glycosylated PD-L1 comprising
a V H domain comprising a CDR H1 with the amino acid sequence of SEQ ID NO: 20, a CDR H2 with the amino acid sequence of SEQ ID NO: 22, and a CDR H3 with the amino acid sequence of SEQ ID NO: 24, or a V H comprising a CDR H1 with the amino acid sequence of SEQ ID NO: 21 a CDR H2 with the amino acid sequence of SEQ ID NO: 23, and a CDR H3 with the amino acid sequence of SEQ ID NO: 25 and
wherein said antibody has a V L domain comprising a CDR L1 with the amino acid sequence of SEQ ID NO: 28, a CDR L2 with the amino acid sequence of SEQ ID NO: 30, and a CDR L3 with the amino acid sequence of SEQ ID NO: 32.
4. An isolated antibody of claim 1 comprising a V H domain encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 2, and/or a V L domain encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 10.
5. The isolated antibody of claim 3 , which has human antibody framework regions or a human constant domain.
6. The isolated antibody of claim 1 , wherein the antibody is at least one selected from the group consisting of a Fab′, a F(ab′)2, a F(ab′)3, a monovalent scFv, a bivalent scFv, a bispecific, and a biparatopic.
7. A composition comprising the antibody of claim 1 in a pharmaceutically acceptable carrier, diluent, excipient, or vehicle.
8. A method of treating a PD-L1 positive cancer in a subject in need, comprising administering to a subject having a PD-L1 positive cancer an effective amount of the isolated antibody of claim 1 .
9. The method of claim 8 , wherein the PD-L1 positive cancer is at least one cancer selected from the group consisting of a breast cancer, a lung cancer, a head & neck cancer, a prostate cancer, an esophageal cancer, a tracheal cancer, a skin cancer, a brain cancer, a liver cancer, a bladder cancer, a stomach cancer, a pancreatic cancer, an ovarian cancer, a uterine cancer, a cervical cancer, a testicular cancer, a colon cancer, a rectal cancer, and a skin cancer.
10. The isolated antibody of claim 1 , wherein the antibody is conjugated to an antineoplastic agent to produce an antibody-drug conjugate (ADC).
11. The isolated antibody of claim 10 , wherein the antibody is chemically conjugated to a maleimide and caproic acid (MC) attachment group, which is chemically conjugated to a cathepsin-cleavable linker, which is chemically conjugated to a paraminobenzoic acid (PAB) spacer, which is chemically conjugated to MMAE, thereby forming the ADC.
12. An ADC comprising a dual function anti-glycPD-L1 antibody chemically coupled to a cytotoxic drug, wherein the anti-glycPD-L1 antibody is that of claim 1 .
13. The isolated antibody of claim 1 , which has human antibody framework regions or a human constant domain.
14. The isolated antibody according to claim 3 , wherein the antibody is at least one selected from the group consisting of a Fab′, a F(ab′)2, a F(ab′)3, a monovalent scFv, a bivalent scFv, a bispecific, and a biparatopic.
15. An isolated antibody of claim 3 having a VH domain comprising the amino acid sequence of SEQ ID NO: 19 and VL domain comprising the amino acid sequence of SEQ ID NO: 27.
16. An isolated antibody of claim 3 comprising a V H domain encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 18, and a V L domain encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 26.
17. A composition comprising the antibody of claim 3 and a pharmaceutically acceptable carrier, diluent, excipient, or vehicle.
18. A method of treating a PD-L1 positive cancer in a subject in need of said treatment, said method comprising administering to the subject an effective amount of the antibody according to claim 3 .
19. The method of claim 18 , wherein the cancer is at least one cancer selected from the group consisting of a hematological cancer, a breast cancer, a lung cancer, a head & neck cancer, a prostate cancer, an esophageal cancer, a tracheal cancer, a skin cancer, a brain cancer, a liver cancer, a bladder cancer, a stomach cancer, a pancreatic cancer, an ovarian cancer, a uterine cancer, a cervical cancer, a testicular cancer, a colon cancer, a rectal cancer, and a skin cancer.
20. A method of assessing PD-L1 glycosylation in a biological sample, said method comprising contacting a biological sample with an antibody according to claim 3 , wherein detection of binding of said antibody indicates that the sample comprises glycosylated PD-L1.
21. The method of claim 20 , wherein the sample is a cell sample from a cancer or tumor of a subject.
22. A method of assessing PD-L1 glycosylation in a biological sample, said method comprising contacting a biological sample with an antibody according to claim 1 , wherein detection of binding of said antibody indicates that the sample comprises glycosylated PD-L1.
23. The method of claim 22 , wherein the sample is a cell sample from a cancer or tumor of a subject.
24. The isolated antibody of claim 1 , wherein the antibody is an IgG antibody, an IgM antibody, or an IgA antibody.
25. The isolated antibody of claim 3 , wherein the antibody is an IgG antibody, an IgM antibody, or an IgA antibody.
26. The isolated antibody of claim 3 , wherein the antibody is conjugated to an antineoplastic agent to produce an antibody-drug conjugate (ADC).
27. The isolated antibody of claim 26 , wherein the antibody is chemically conjugated to a maleimide and caproic acid (MC) attachment group, which is chemically conjugated to a cathepsin-cleavable linker, which is chemically conjugated to a paraminobenzoic acid (PAB) spacer, which is chemically conjugated to MMAE, thereby forming the ADC.
28. An ADC comprising a dual function anti-glycPD-L1 antibody chemically coupled to a cytotoxic drug, wherein the anti-glycPD-L1 antibody is that of claim 3 .