NZ727772B2 - Covalently bonded diabodies having immunoreactivity with pd-1 and lag-3, and methods of use thereof - Google Patents

Abstract

The present invention is directed to bi-specific diabodies that comprise two or more polypeptide chains and which possess at least one Epitope-Binding Site that is immunospecific for an epitope of PD-1 and at least one Epitope-Binding Site that is immunospecific for an epitope of LAG-3 (i.e., a "PD-I x LAG-3 bi-specific diabody"). More preferably, the present invention is directed to bi-specific diabodies that comprise four polypeptide chains and which possess two Epitope-Binding Sites that are immunospecific for one (or two) epitope(s) of PD-1 and two Epitope-Binding Site that are immunospecific for one (or two) epitope(s) of LAG-3 (i.e., a "PD-1 x LAG-3 bi-specific, tetra-valent diabody").

Claims (20)

What Is Claimed Is:

1.Claim 1. A bi-specific Fc diabody capable of immunospecific binding to an epitope of PD-1 and to an epitope of LAG-3, wherein said diabody comprises four polypeptide chains, each having an amino terminus and a carboxy terminus, and n: (A) said first and second polypeptide chains are covalently bonded to one another, said first and third polypeptide chains are covalently bonded to one another, and said third and fourth polypeptide chains are covalently bonded to one another; (B) said first and third polypeptide chains of said diabody each comprise, in the N-terminal to C-terminal direction, a Light Chain Variable Domain of an antibody that is immunospecific for PD-1 or LAG-3, a Heavy Chain Variable Domain of an antibody that is immunospecific for LAG-3 or PD- 1, and a CH2-CH3 Domain, wherein said Light Chain Variable Domains and said Heavy Chain Variable Domains are incapable of associating to form an Epitope-Binding Site capable of g an epitope of PD-1 or an epitope of LAG-3; and (C) said second and fourth polypeptide chains of said diabody each comprise, in the N-terminal to C-terminal direction, a Light Chain Variable Domain of an dy that is immunospecific for PD-1 or LAG-3, a Heavy Chain Variable Domain of an dy that is immunospecific for LAG-3 or PD- 1, wherein said Light Chain le Domains and said Heavy Chain Variable Domains are incapable of associating to form an e-Binding Site capable of binding an epitope of PD-1 or an epitope of LAG-3; and wherein: (I) (1) said Light Chain le Domain of said first polypeptide chain and said Heavy Chain Variable Domain of said second polypeptide chain associate to form a first Epitope-Binding Site and said Heavy Chain Variable Domain of said first polypeptide chain and said Light Chain Variable Domain of said second ptide chain associate to form a second Epitope-Binding Site; and (2) said Light Chain Variable Domain of said third polypeptide chain and said Heavy Chain Variable Domain of said fourth polypeptide chain associate to form a third Epitope-Binding Site and said Heavy Chain Variable Domain of said third polypeptide chain and said Light Chain Variable Domain of said fourth ptide chain associate to form a fourth Epitope-Binding Site; wherein two of said formed Epitope-Binding Sites are capable of immunospecifically binding to an epitope of PD-1 and two of said formed Epitope-Binding Sites are capable of immunospecifically binding to an epitope of LAG-3; and II. said CH2-CH3 Domains of said first and third polypeptide chains associate to form an Fc Domain.

2.Claim 2. The bi-specific Fc diabody of claim 1, wherein: (A) said first and third polypeptide chains of said diabody each comprise, in the N-terminal to C-terminal direction, a Light Chain le Domain of an antibody that is immunospecific for PD-1 or LAG-3, a Heavy Chain Variable Domain of an antibody that is immunospecific for LAG-3 or PD- 1, a Heterodimer-Promoting Domain and a CH2-CH3 Domain, wherein said Light Chain Variable Domains and said Heavy Chain le Domains are incapable of associating to form an Epitope-Binding Site capable of binding an epitope of PD-1 or an epitope of LAG-3; and (B) said second and said fourth polypeptide chains of said diabody each comprise, in the N-terminal to C-terminal direction, a Light Chain le Domain of an antibody that is immunospecific for PD-1 or LAG-3, a Heavy Chain Variable Domain of an antibody that is immunospecific for LAG-3 or PD-1, and a dimer-Promoting , wherein said Light Chain Variable s and said Heavy Chain Variable Domains are incapable of associating to form an Epitope-Binding Site capable of binding an epitope of PD-1 or an epitope of LAG-3; wherein said Heterodimer-Promoting Domain of said first and third polypeptide chains differs from said Heterodimer-Promoting Domain of said second and fourth polypeptide chains.

3.Claim 3. The bi-specific Fc diabody of claim 2, wherein: (1) said Heterodimer-Promoting Domain of said first and third polypeptide chains comprises the amino acid sequence of SEQ ID NO:14 and said Heterodimer-Promoting Domain of said second and fourth ptide chains has the amino acid ce of SEQ ID NO:15; or (2) said Heterodimer-Promoting Domain of said first and third ptide chains comprises the amino acid sequence of SEQ ID NO:15 and said Heterodimer-Promoting Domain of said second and fourth polypeptide chains has the amino acid sequence of SEQ ID NO:14; or (3) said Heterodimer-Promoting Domain of said first and third polypeptide chains comprises the amino acid sequence of SEQ ID NO:16 and said dimer-Promoting Domain of said second and fourth polypeptide chains has the amino acid sequence of SEQ ID NO:17; or (4) said Heterodimer-Promoting Domain of said first and third polypeptide chains comprises the amino acid ce of SEQ ID NO:17 and said Heterodimer-Promoting Domain of said second and fourth polypeptide chains has the amino acid sequence of SEQ ID NO:16.

4.Claim 4. The bi-specific Fc diabody of any one of claims 2-3, wherein said Heterodimer- Promoting Domain of said first polypeptide chain comprises SEQ ID NO:14 and said Heterodimer-Promoting Domain of said second polypeptide chain comprises SEQ ID NO:15.

5.Claim 5. The bi-specific Fc diabody of any one of claims 2-3, wherein said Heterodimer- Promoting Domain of said first polypeptide chain comprises SEQ ID NO:16 and said Heterodimer-Promoting Domain of said second polypeptide chain comprises SEQ ID NO:17.

6.Claim 6. The bi-specific Fc diabody of any one of claims 2-5, wherein said Heavy Chain Variable Domain is separated from said Heterodimer-Promoting Domain in each of said first, second, third and fourth polypeptide chains by a linker comprising the amino acid sequence of SEQ ID NO:18.

7.Claim 7. The cific Fc diabody of any one of claims 1-6, wherein said CH2-CH3 Domains of said first and third ptide chains each comprise the amino acid sequence of SEQ ID NO:24.

8.Claim 8. The bi-specific Fc diabody of any one of claims 1-7, wherein said Heavy Chain Variable Domain of an antibody that is immunospecific for LAG-3 comprises the amino acid sequence of SEQ ID NO:11, and wherein said Light Chain Variable Domain of an antibody that is immunospecific for LAG-3 comprises the amino acid sequence of SEQ ID NO:12.

9.Claim 9. The bi-specific Fc diabody of any one of claims 1-8, wherein said Heavy Chain Variable Domain of an antibody that is immunospecific for PD-1 comprises the amino acid sequence of SEQ ID NO:2, and wherein said Light Chain Variable Domain of an antibody that is immunospecific for PD-1 comprises the amino acid ce of SEQ ID NO:3.

10.Claim 10. The bi-specific Fc diabody of any one of claims 1-9, wherein: (A) said first and third polypeptide chains each comprise a Light Chain Variable Domain of an antibody that is immunospecific for PD-1 and a Heavy Chain Variable Domain of an antibody that is specific for LAG-3; and (B) said second and fourth polypeptide chains each comprise a Light Chain Variable Domain of an dy that is immunospecific for LAG-3 and a Heavy Chain Variable Domain of an antibody that is immunospecific for PD-1.

11.Claim 11. The bi-specific Fc y of any one of claims 1-9, wherein: (A) said first and third polypeptide chains each comprise a Light Chain Variable Domain of an antibody that is immunospecific for LAG-3 and a Heavy Chain le Domain of an antibody that is immunospecific for PD-1; (B) said second and fourth polypeptide chains each comprise a Light Chain Variable Domain of an antibody that is specific for PD-1 and a Heavy Chain le Domain of an antibody that is immunospecific for LAG-3.

12.Claim 12. A pharmaceutical composition that comprises an effective amount of the bi-specific Fc diabody of any one of claims 1-11, and a pharmaceutically acceptable r.

13.Claim 13. The pharmaceutical composition of claim 12, wherein said effective amount of said bi-specific Fc diabody is an amount effective to treat cancer in a recipient individual in need of such treatment.

14.Claim 14. The ceutical ition of claim 13, wherein said cancer is an adrenal gland cancer, an ssociated 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 desmoplastic small round cell tumor, an ependymoma, a Ewing’s tumor, an extraskeletal myxoid chondrosarcoma, a gallbladder or bile duct cancer, gastric cancer, a gestational trophoblastic disease, a germ cell tumor, a head and neck cancer, cellular carcinoma, an islet cell tumor, a Kaposi’s sarcoma, a kidney cancer, a leukemia, a liposarcoma/malignant lipomatous tumor, a liver cancer, a lymphoma, a lung cancer, a oblastoma, a melanoma, a meningioma, a le 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 pheochromocytoma, a pituitary tumor, a prostate cancer, a posterior uveal ma, a renal metastatic , a rhabdoid tumor, a rhabdomyosarcoma, a sarcoma, a skin cancer, a soft-tissue sarcoma, a squamous cell cancer, a stomach cancer, a al sarcoma, a testicular cancer, a thymic carcinoma, a thymoma, a thyroid metastatic cancer, or a uterine cancer.

15.Claim 15. The pharmaceutical composition of claim 12, wherein said effective amount of said cific Fc diabody is an amount effective to treat a disease associated with the presence of a pathogen in a recipient individual in need of such treatment.

16.Claim 16. The ceutical composition of claim 15, wherein said pathogen is a bacterium, a fungus, a protozoan, or a virus.

17.Claim 17. Use of the bi-specific Fc diabody of any of claims 1-11, or the pharmaceutical composition of any of claims 13-14, in the manufacture of a medicament for treating cancer.

18.Claim 18. Use of the bi-specific Fc y of any of claims 1-11, or the ceutical composition of any of claim 15-16, in the manufacture of a ment for treating a disease associated with the presence of a pathogen.

19.Claim 19. The bi-specific Fc diabody of any one of claims 1-11, or the pharmaceutical composition of any one of claims 12-14, for use in treating cancer in an individual in need thereof.

20.Claim 20. The bi-specific Fc y of any one of claims 1-11, or the pharmaceutical composition of any one of claims 12, 15 or 16, for use in treating a disease associated with the presence of a pathogen in an individual in need thereof. Diabody—Type Binding Site Binds First E a": \