Multi-chain chimeric polypeptides and uses thereof

Abstract

To provide multi-chain chimeric polypeptides.SOLUTION: There are provided herein multi-chain chimeric polypeptides that include: (a) a first chimeric polypeptide including a first target-binding domain, a soluble tissue factor domain, and a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide including a second domain of a pair of affinity domains and a second target-binding domain, where the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains. Also provided herein are methods of using these multi-chain chimeric polypeptides, and nucleic acids encoding these multi-chain chimeric polypeptides.SELECTED DRAWING: Figure 1

Claims

1. (a)(i) a first target binding domain that specifically binds TGF-β, wherein the first target binding domain comprises a sequence at least 90% identical to SEQ ID NO: 85; (ii) a soluble tissue factor domain comprising a sequence at least 90% identical to SEQ ID NO: 1; (iii) a first domain of a pair of affinity domains comprising a sequence at least 90% identical to SEQ ID NO: 14; and a first chimeric polypeptide comprising: (b) (i) a second domain of the pair of affinity domains comprising a sequence at least 90% identical to SEQ ID NO: 28; (ii) a second target binding domain that specifically binds to TGF-β, comprising a sequence at least 90% identical to SEQ ID NO: 85; and a second chimeric polypeptide comprising wherein the first chimeric polypeptide and the second chimeric polypeptide are associated via binding between the first domain and the second domain of the pair of affinity domains.

2. The multi-chain chimeric polypeptide of claim 1 , wherein the first target binding domain and the soluble tissue factor domain are immediately adjacent to each other within the first chimeric polypeptide.

3. The multi-chain chimeric polypeptide of claim 1, wherein the first chimeric polypeptide further comprises a linker sequence between the first target binding domain and the soluble tissue factor domain within the first chimeric polypeptide.

4. The multi-chain chimeric polypeptide of any one of claims 1 to 3, wherein the soluble tissue factor domain and the first domain of the pair of affinity domains are immediately adjacent to each other within the first chimeric polypeptide.

5. The multi-chain chimeric polypeptide of any one of claims 1 to 3, wherein the first chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain in the first chimeric polypeptide and the first domain of the pair of affinity domains.

6. The multi-chain chimeric polypeptide of any one of claims 1 to 5, wherein the second domain and the second target binding domain of the pair of affinity domains are immediately adjacent to each other within the second chimeric polypeptide.

7. The multi-chain chimeric polypeptide of any one of claims 1 to 5, wherein the second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains within the second chimeric polypeptide and the second target binding domain.

8. The multi-chain chimeric polypeptide of any one of claims 1 to 7, wherein the first chimeric polypeptide further comprises one or more additional target binding domain(s).

9. The multi-chain chimeric polypeptide of any one of claims 1 to 8, wherein the second chimeric polypeptide further comprises one or more additional target binding domains.

10. the first target binding domain comprises a sequence at least 95% identical to SEQ ID NO: 85; the soluble tissue factor domain comprises a sequence at least 95% identical to SEQ ID NO: 1; the first domain of the pair of affinity domains comprises a sequence at least 95% identical to SEQ ID NO: 14; The second domain of the pair of affinity domains comprises a sequence that is at least 95% identical to SEQ ID NO: 28; and The multi-chain chimeric polypeptide of claim 1 , wherein the second target binding domain comprises a sequence that is at least 95% identical to SEQ ID NO:

85.

11. the first target binding domain comprises a sequence at least 99% identical to SEQ ID NO: 85; the soluble tissue factor domain comprises a sequence at least 99% identical to SEQ ID NO: 1; the first domain of the pair of affinity domains comprises a sequence at least 99% identical to SEQ ID NO: 14; The second domain of the pair of affinity domains comprises a sequence that is at least 99% identical to SEQ ID NO: 28; and The multi-chain chimeric polypeptide of claim 10 , wherein the second target binding domain comprises a sequence that is at least 99% identical to SEQ ID NO:

85.

12. the first target binding domain comprises SEQ ID NO: 85; the soluble tissue factor domain comprises SEQ ID NO: 1; the first domain of the pair of affinity domains comprises SEQ ID NO: 14; the second domain of the pair of affinity domains comprises SEQ ID NO: 28; and the second target binding domain comprises SEQ ID NO: 85; The multi-chain chimeric polypeptide of claim 11.

13. the first chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 133; and the second chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 177; The multi-chain chimeric polypeptide according to any one of claims 1 to 7.

14. the first chimeric polypeptide comprises a sequence at least 95% identical to SEQ ID NO: 133; and the second chimeric polypeptide comprises a sequence at least 95% identical to SEQ ID NO: 177; The multi-chain chimeric polypeptide of claim 13.

15. the first chimeric polypeptide comprises a sequence at least 99% identical to SEQ ID NO: 133; and the second chimeric polypeptide comprises a sequence at least 99% identical to SEQ ID NO: 177; The multi-chain chimeric polypeptide of claim 14.

16. the first chimeric polypeptide comprises SEQ ID NO: 133, and the second chimeric polypeptide comprises SEQ ID NO: 177; The multi-chain chimeric polypeptide of claim 15.

17. A nucleic acid encoding one or more of the multi-chain chimeric polypeptides of claims 1 to 16.

18. A pharmaceutical composition for stimulating immune cells or for inducing or increasing the proliferation of immune cells, comprising an effective amount of any of the multi-chain chimeric polypeptides of claims 1 to 16.

19. 19. The pharmaceutical composition of claim 18, wherein the immune cells are selected from the group consisting of immature thymocytes, peripheral blood lymphocytes, naive T cells, multipotent Th cell precursors, lymphoid progenitor cells, Treg cells, Th17 cells, Th22 cells, Th9 cells, Th2 cells, Th1 cells, Th3 cells, γδ T cells, αβ T cells, tumor-infiltrating T cells, CD8+ T cells, CD4+ T cells, natural killer T cells, mast cells, macrophages, neutrophils, dendritic cells, basophils, eosinophils, and natural killer cells.

20. 20. The pharmaceutical composition of claim 18 or 19, wherein the immune cells have been previously genetically modified to express a chimeric antigen receptor or a recombinant T cell receptor.

21. An effective amount of any of the multi-chain chimeric polypeptides of claims 1 to 16. A pharmaceutical composition for inducing differentiation of immune cells into memory immune cells or memory-like immune cells, comprising:

22. 17. A pharmaceutical composition for killing cancer cells, infected cells, or senescent cells in a subject in need thereof, comprising an effective amount of any of the multi-chain chimeric polypeptides of claims 1 to 16.

23. A pharmaceutical composition for treating a subject in need thereof, comprising a therapeutically effective amount of any of the multi-chain chimeric polypeptides of claims 1 to 16.

24. 24. The pharmaceutical composition of claim 23, wherein the subject has been identified or diagnosed as having cancer.

25. 25. The pharmaceutical composition of claim 24, wherein the cancer is selected from the group consisting of solid tumors, hematological tumors, sarcoma, osteosarcoma, glioblastoma, neuroblastoma, melanoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, B-cell neoplasms, multiple myeloma, B-cell lymphoma, B-cell non-Hodgkin's lymphoma, Hodgkin's lymphoma, chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), acute lymphocytic leukemia (ALL), myelodysplastic syndromes (MDS), cutaneous T-cell lymphoma, retinoblastoma, gastric cancer, urothelial carcinoma, lung cancer, renal cell carcinoma, gastroesophageal cancer, pancreatic cancer, prostate cancer, breast cancer, colorectal cancer, ovarian cancer, non-small cell lung cancer, squamous cell head and neck cancer, endometrial cancer, cervical cancer, liver cancer, and hepatocellular carcinoma.

26. A pharmaceutical composition for treating a subject identified or diagnosed as having an infectious disease, comprising a therapeutically effective amount of any of the multi-chain chimeric polypeptides of claims 1-16.

27. 27. The pharmaceutical composition of claim 26, wherein the infectious disease is infection with cytomegalovirus (CMV).

28. 27. The pharmaceutical composition of claim 26, wherein the infectious disease is infection with human immunodeficiency virus (HIV).

29. 27. The pharmaceutical composition of claim 26, wherein the infectious disease is an infection with influenza.

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