NZ778489B2

NZ778489B2 - PD-1-binding molecules and methods of use thereof

Info

Publication number: NZ778489B2
Application number: NZ778489A
Authority: NZ
Inventors: Ezio Bonvini; Leslie S Johnson; Scott Koenig; Motte Mohs Ross La; Paul A Moore; Kalpana Shah; Douglas H Smith
Original assignee: Macrogenics Inc
Filing date: 2016-07-28
Publication date: 2025-01-28

Abstract

The present invention is directed to selected anti-PD-1 antibodies capable of binding to both cynomolgus monkey PD-1 and to human PD-1 : PD-1 mAb 1, PD-1 mAb 2, PD-1 mAb 3, PD-1 mAb 4, PD-1 mAb 5, PD-1 mAb 6, PD-1 mAb 7, PD-1 mAb 8, PD-1 mAb 9, PD-1 mAb 10, PD-1 mAb 11, PD-1 mAb 12, PD-1 mAb 13, PD 1 mAb 14, or PD-1 mAb 15, and to humanized and chimeric versions of such antibodies. The invention additionally pertains to PD-1 -binding molecules that comprise PD-1 binding fragments of such anti-PD-1 antibodies, immunocongugates, and to bispecific molecules, including diabodies, BiTEs, bispecific antibodies, etc., that comprise (i) such PD-1 -binding fragments, and (ii) a domain capable of binding an epitope of a molecule involved in regulating an immune check point present on the surface of an immune cells. The present invention also pertains to methods of using molecules that bind PD-1 for stimulating immune responses, as well as methods of detecting PD-1.

Claims

What is Claimed is:

1. Claim 1. A bispecific binding molecule capable of binding to PD-1 and LAG-3, wherein the bispecific binding molecule comprises: (I) a PD-1 binding domain comprising a Variable Heavy Chain Domain and a Variable Light Chain Domain, wherein said Variable Heavy Chain Domain comprises a CDR 1 Domain, a CDR 2 Domain and a CDR 3 Domain, and said Variable Light Chain Domain comprises a CDR 1 Domain, a CDR 2 Domain, and a CDR 3 Domain, wherein: L L L (A) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the H H H Heavy Chain CDRs of PD-1 mAb 7, and respectively comprise the amino acid sequences: SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the L L L Light Chain CDRs of PD-1 mAb 7, and, respectively comprise the amino acid sequences: SEQ ID NO:144, SEQ ID NO:145, and SEQ ID NO:146; (B) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the H H H Heavy Chain CDRs of hPD-1 mAb 7(1.2), and respectively comprise the amino acid sequences: SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the L L L Light Chain CDRs of hPD-1 mAb 7(1.2), and, respectively comprise the amino acid sequences: SEQ ID NO:157, SEQ ID NO:145, and SEQ ID NO:146; (C) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the H H H Heavy Chain CDRs of hPD-1 mAb 7(1.3), and respectively comprise the amino acid sequences: SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the L L L Light Chain CDRs of hPD-1 mAb 7(1.3), and, respectively comprise the amino acid sequences: SEQ ID NO:157, SEQ ID NO:158, and SEQ ID NO:146; and (II) a LAG-3 binding domain comprising a Variable Heavy Chain Domain and a Variable Light Chain Domain, wherein the Variable Heavy Chain Domain of the LAGbinding domain comprises a CDR 1 Domain, a CDR 2 Domain, and a CDR 3 Domain, and the Variable Light Chain Domain of the LAGbinding domain comprises a CDR 1 Domain, a CDRL2 Domain, and a CDRL3 Domain.

2. Claim 2. The bispecific binding molecule of claim 1, wherein: (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the H H H Heavy Chain CDRs of hPD-1 mAb 7(1.2), and respectively comprise the amino acid sequences: SEQ ID NO:139, SEQ ID NO:140, and SEQ ID NO:141; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain are the L L L Light Chain CDRs of hPD-1 mAb 7(1.2), and, respectively comprise the amino acid sequences: SEQ ID NO:157, SEQ ID NO:145, and SEQ ID NO:146.

3. Claim 3. The bispecific binding molecule of claim 1 or 2, wherein said molecule comprises a Heavy Chain Variable Domain that comprises the amino acid sequence of SEQ ID NO:147.

4. Claim 4. The bispecific binding molecule of any one of claims 1-3, wherein said molecule comprises a Light Chain Variable Domain that comprises the amino acid sequence of SEQ ID NO:151, SEQ ID NO:153, or SEQ ID NO:155.

5. Claim 5. The bispecific binding molecule of any one of claims 1-4, wherein: (A) said molecule comprises a Heavy Chain Variable Domain that comprises the amino acid sequence of SEQ ID NO:147; or (B) said molecule comprises a Light Chain Variable Domain that comprises the amino acid sequence of SEQ ID NO:153.

6. Claim 6. The bispecific binding molecule of any one of claims 1-5 wherein said LAG-3 epitope-binding site comprises: (A) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the H H H Variable Heavy Chain of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:42, SEQ ID NO:43, and SEQ ID NO:44; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the L L L Variable Light Chain of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:46, SEQ ID NO:47, and SEQ ID NO:48; (B) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the H H H Variable Heavy Chain of hLAG-3 mAb 1 VH1, and respectively comprise the amino acid sequences: SEQ ID NO:42, SEQ ID NO:43, and SEQ ID NO:44; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the L L L Variable Light Chain of LAG-3 mAb 1 VL4, and respectively comprise the amino acid sequences: SEQ ID NO:55, SEQ ID NO:47, and SEQ ID NO:48; (C) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the H H H Variable Heavy Chain of LAG-3 mAb 6, and respectively comprise the amino acid sequences: SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the L L L Variable Light Chain of LAG-3 mAb 6, and respectively comprise the amino acid sequences: SEQ ID NO:61, SEQ ID NO:62, and SEQ ID NO:63; (D) (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the H H H Variable Heavy Chain of hLAG-3 mAb 6 VH1, and respectively comprise the amino acid sequences: SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59; and (2) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the L L L Variable Light Chain of hLAG-3 mAb 6 VL1, and respectively comprise the amino acid sequences: SEQ ID NO:298, SEQ ID NO:62, and SEQ ID NO:63.

7. Claim 7. The bispecific binding molecule of any one of claims 1-5, wherein said LAG-3 epitope-binding site comprises: (1) the CDR 1 Domain, CDR 2 Domain, and CDR 3 Domain of the Variable H H H Heavy Chain of hLAG-3 mAb 6 VH1, and respectively comprise the amino acid sequences: SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59; and (2) the CDRL1 Domain, CDRL2 Domain, and CDRL3 Domain of the Variable Light Chain of hLAG-3 mAb 6 VL1/VL2 and respectively comprise the amino acid sequences: SEQ ID NO:298, SEQ ID NO:62, and SEQ ID NO:63.

8. Claim 8. The bispecific binding molecule of any one of claims 1-5, wherein said LAG-3 epitope-binding site comprises: (1) the Variable Heavy Chain of hLAG-3 mAb 6 VH1 (SEQ ID NO:294) and (2) the Variable Light Chain of hLAG-3 mAb 6 VL1 (SEQ ID NO:296).

9. Claim 9. The bispecific binding molecule of any one of claims 1-8, wherein said molecule (A) a diabody, said diabody being a covalently bonded complex that comprises four or five polypeptide chains; or (B) a bispecific antibody.

10. Claim 10. The bispecific binding molecule of any one of claims 1-9, wherein said molecule comprises an Fc Region.

11. Claim 11. The bispecific binding molecule of claim 10, wherein said Fc Region is of the IgG1, IgG2, IgG3, or IgG4 isotype.

12. Claim 12. The bispecific binding molecule of claim 11, wherein said molecule or said antibody further comprises a Hinge Domain.

13. Claim 13. The bispecific binding molecule of claim 12, wherein said Fc Region and said Hinge Doman are of the IgG4 isotype, and wherein said Hinge Domain comprises a stabilizing mutation.

14. Claim 14. The bispecific binding molecule of any one of claims 10-13, wherein said Fc Region is a variant Fc Region that comprises: (a) one or more amino acid modifications that reduces the affinity of the variant Fc Region for an Fc?R; and/or (b) one or more amino acid modifications that enhances the serum half-life of the variant Fc Region.

15. Claim 15. The bispecific binding molecule of claim 14, wherein said one or more amino acid modifications that reduces the affinity of the variant Fc Region for an Fc?R comprise the substitution of L234A; L235A; or L234A and L235A, wherein said numbering is that of the EU index as in Kabat.

16. Claim 16. The bispecific binding molecule of claim 14 or 15, wherein said one or more amino acid modifications that that enhances the serum half-life of the variant Fc Region comprise the substitution of M252Y; M252Y and S254T; M252Y and T256E; M252Y, S254T and T256E; or K288D and H435K, wherein said numbering is that of the EU index as in Kabat.

17. Claim 17. The bispecific binding molecule of any one of claims 1-16, wherein said molecule is a diabody comprising: (a) SEQ ID NO:267, wherein X is Ala; X is Tyr; X is Thr; X is Glu, and 1 2 3 4 SEQ ID NO:268; or (b) SEQ ID NO:267, wherein X is Gly; X is Tyr; X is Thr; X is Glu, and 1 2 3 4 SEQ ID NO:268; or (c) SEQ ID NO:267, wherein X is Gly; X is Met; X is Ser; X is Thr, and 1 2 3 4 SEQ ID NO:268; or (d) SEQ ID NOs:269 and 270; or (e) SEQ ID NOs:271 and 272; or (f) SEQ ID NOs:273, 274, 275, and 276; or (g) SEQ ID NOs:277, 278, 279, and 280; or (h) SEQ ID NOs:281, 282, and 283; or (i) SEQ ID NOs:290 and 291; or (j) SEQ ID NOs:292 and 293.

18. Claim 18. The bispecific binding molecule of claim 17, wherein said molecule is a diabody comprising SEQ ID NOs:290 and 291.

19. Claim 19. A composition comprising: (A) the bispecific binding molecule of any one of claims 1-18; and (B) a pharmaceutically acceptable carrier.

20. Claim 20. The bispecific binding molecule of any one of claims 1-18, wherein said molecule is detectably labeled and is used in the detection of PD-1.

21. Claim 21. Use of the bispecific binding molecule of any one of claims 1-18, for the manufacture of a medicament for the treatment of a disease or condition associated with a suppressed immune system.

22. Claim 22. The use of claim 21, wherein the disease or condition is cancer or an infection.

23. Claim 23. The use of claim 22, wherein said cancer is characterized by the presence of a cancer cell selected from the group consisting of a cell of: an adrenal gland tumor, an AIDS-associated cancer, an alveolar soft part sarcoma, an astrocytic tumor, bladder cancer, bone cancer, a brain and spinal cord cancer, a metastatic brain tumor, a breast cancer, a carotid body tumors, a cervical cancer, a chondrosarcoma, a chordoma, a chromophobe renal cell carcinoma, a clear cell carcinoma, a colon cancer, a colorectal cancer, a cutaneous benign fibrous histiocytoma, a desmoplastic small round cell tumor, an ependymoma, a Ewing’s tumor, an extraskeletal myxoid chondrosarcoma, a fibrogenesis imperfecta ossium, a fibrous dysplasia of the bone, a gallbladder or bile duct cancer, gastric cancer, a gestational trophoblastic disease, a germ cell tumor, a head and neck cancer, hepatocellular carcinoma, an islet cell tumor, a Kaposi’s Sarcoma, a kidney cancer, a leukemia, a lipoma/benign lipomatous tumor, a liposarcoma/malignant lipomatous tumor, a liver cancer, a lymphoma, a lung cancer, a medulloblastoma, a melanoma, a meningioma, a multiple endocrine neoplasia, a multiple myeloma, a myelodysplastic syndrome, a neuroblastoma, a neuroendocrine tumors, an ovarian cancer, a pancreatic cancer, a papillary thyroid carcinoma, a parathyroid tumor, a pediatric cancer, a peripheral nerve sheath tumor, a phaeochromocytoma, a pituitary tumor, a prostate cancer, a posterious uveal melanoma, a rare hematologic disorder, a renal metastatic cancer, a rhabdoid tumor, a rhabdomysarcoma, a sarcoma, a skin cancer, a soft-tissue sarcoma, a squamous cell cancer, a stomach cancer, a synovial sarcoma, a testicular cancer, a thymic carcinoma, a thymoma, a thyroid metastatic cancer, and a uterine cancer.

24. Claim 24. A nucleic acid expression vector encoding the bispecific binding molecule of any one of claims 1-18.