阅读说明：本技术 抗体细胞因子移植蛋白和用于治疗癌症的方法 (Antibody cytokine transplantation proteins and methods for treating cancer ) 是由 J·迪恩 Y·迪亚兹-德-杜拉纳 M·迪多纳托 C·菲利皮 G·斯普拉贡 于 2018-05-22 设计创作，主要内容包括：本披露提供了移植到抗体的CDR序列中的IL2,与临床上已知和使用的分子相比,其具有优选的治疗特性。特别地,所提供的抗体细胞因子移植蛋白组合物增加或维持CD8+T效应细胞,同时降低Treg细胞的活性。另外,所提供的组合物比重组人IL2配制品例如<Image he="68" wi="298" file="DDA0002284083990000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>传达了改善的半衰期、稳定性和可生产性。(The present disclosure provides IL2 grafted into the CDR sequences of antibodies with preferred therapeutic properties compared to clinically known and used molecules. In particular, the provided antibody cytokine transplantation protein compositions increase or maintain CD8+ T effector cells while decreasing Treg cell activity. In addition, the compositions provided are more recombinant than recombinant humansIL2 formulations such as Conveying improved half-life, stability and producibility.)

1. An antibody cytokine graft protein comprising:

(a) a heavy chain variable region (VH) comprising Complementarity Determining Regions (CDRs) HCDR1, HCDR2, HCDR 3; and

(b) a light chain variable region (VL) comprising LCDR1, LCDR2, LCDR 3; and

(c) an interleukin 2(IL2) molecule grafted into the CDR of the VH or the VL.

2. The antibody cytokine graft protein of claim 1, wherein the IL2 molecule is grafted into the heavy chain CDRs.

3. The antibody cytokine graft protein of claim 2, wherein the heavy chain CDRs are selected from the group consisting of complementarity determining region 1(HCDR1), complementarity determining region 2(HCDR2) and complementarity determining region 3(HCDR 3).

4. The antibody cytokine graft protein of claim 3, wherein the IL2 molecule is engrafted in HCDR 1.

5. The antibody cytokine graft protein of claim 1, wherein the IL2 molecule is grafted into the light chain CDRs.

6. The antibody cytokine graft protein of claim 5, wherein the light chain CDR is selected from complementarity determining region 1(LCDR1), complementarity determining region 2(LCDR2), and complementarity determining region 3(LCDR 3).

7. The antibody cytokine graft protein of any one of claims 1-6, wherein the IL2 molecule contains a mutation that reduces the affinity of the IL2 molecule for a high affinity IL2 receptor.

8. The antibody cytokine graft protein of any one of claims 1-7, wherein the anti-cytokine isThe somatic cell factor transplanting protein is compared with the recombinant IL2 or

9. The antibody cytokine graft protein of any one of claims 1-8, wherein the antibody cytokine graft protein is more recombinant IL2 or

10. The antibody cytokine graft protein of any one of claims 1-9, wherein the antibody cytokine graft protein is more recombinant IL2 or

11. The antibody cytokine graft protein of any one of claims 1-10, wherein the antibody cytokine graft protein is more native than IL2 or

12. The antibody cytokine graft protein of any one of claims 1-11, wherein the IL2 molecule consists of SEQ ID No. 4.

13. The antibody cytokine graft protein of any one of claims 1-11, wherein the IL2 molecule consists of SEQ ID No. 6.

14. The antibody cytokine graft protein of any one of claims 1-13, further comprising an IgG class antibody heavy chain.

15. The antibody cytokine graft protein of claim 14, wherein the IgG class heavy chain is selected from the group consisting of IgG1, IgG2, and IgG 4.

16. The antibody cytokine graft protein of any one of claims 1-15, wherein the binding specificity of the CDRs to a target is reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99% or 100% by the grafted IL2 molecule.

17. The antibody cytokine graft protein of any one of claims 1-15, wherein the binding specificity of the CDRs to a target is reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99% or 100% in the presence of the grafted IL2 molecule.

18. The antibody cytokine graft protein of any one of claims 1-17, wherein the binding specificity of the CDRs is different from the binding specificity of the IL2 molecule.

19. The antibody cytokine graft protein of any one of claims 1-18, wherein the binding specificity of the CDRs is for a non-human antigen.

20. The antibody cytokine graft protein of claim 19, wherein the non-human antigen is a virus.

21. The antibody cytokine graft protein of claim 20, wherein the virus is Respiratory Syncytial Virus (RSV).

22. The antibody cytokine graft of claim 21, wherein the RSV is selected from RSV subgroup a and RSV subgroup B.

23. The antibody cytokine graft protein of any one of claims 1-22, wherein the antibody scaffold portion of the antibody cytokine graft protein is humanized or human.

24. An antibody cytokine graft protein comprising:

(i) a heavy chain variable region comprising (a) HCDR1 of SEQ ID NO:13, (b) HCDR2 of SEQ ID NO:14, (c) HCDR3 of SEQ ID NO:15, and a light chain variable region comprising: (d) LCDR1 of SEQ ID NO. 29, (e) LCDR2 of SEQ ID NO. 30, and (f) LCDR3 of SEQ ID NO. 31; or

(ii) A heavy chain variable region comprising (a) HCDR1 of SEQ ID NO:45, (b) HCDR2 of SEQ ID NO:46, (c) HCDR3 of SEQ ID NO: 47; and a light chain variable region comprising: (d) LCDR1 of SEQ ID NO:61, (e) LCDR2 of SEQ ID NO:62, and (f) LCDR3 of SEQ ID NO: 63.

25. An antibody cytokine graft protein comprising:

(i) a heavy chain variable region (VH) comprising SEQ ID NO 19 and a light chain variable region (VL) comprising SEQ ID NO 35; or

(ii) The heavy chain variable region (VH) comprising SEQ ID NO 51 and the light chain variable region (VL) comprising SEQ ID NO 67.

26. The antibody cytokine graft protein of any one of claims 1-25, further comprising a modified Fc region corresponding to reduced effector function.

27. The antibody cytokine graft protein of claim 26, wherein the modified Fc region comprises a mutation selected from one or more of D265A, P329A, P329G, N297A, L234A and L235A.

28. The antibody cytokine graft protein of claim 27, wherein the modified Fc region comprises a combination of mutations selected from one or more of D265A/P329A, D265A/N297A, L234/L235A, P329A/L234A/L235A, and P329G/L234A/L235A.

29. An antibody cytokine transplantation protein comprising HCDR1 of SEQ ID NO 13, HCDR2 of SEQ ID NO 14, HCDR3 of SEQ ID NO 15, LCDR1 of SEQ ID NO 29, LCDR2 of SEQ ID NO 30, LCDR3 of SEQ ID NO 31, a modified Fc region containing said mutation D265A/P329A, when combined with

30. An antibody cytokine transplantation protein comprising HCDR1 of SEQ ID NO 45, HCDR2 of SEQ ID NO 46, HCDR3 of SEQ ID NO 47, LCDR1 of SEQ ID NO 61, LCDR2 of SEQ ID NO 62, LCDR3 of SEQ ID NO 63, a modified Fc region containing said mutation D265A/P329A, wherein when combined with said mutation

31. An isolated nucleic acid encoding an antibody cytokine graft protein, comprising: (i) the heavy chain of SEQ ID NO 22 and the light chain of SEQ ID NO 38; or (ii) the heavy chain of SEQ ID NO:54 and the light chain of SEQ ID NO: 70.

32. A recombinant host cell suitable for producing an antibody cytokine graft protein, comprising the nucleic acid of claim 31 encoding the heavy and light chain polypeptides of the antibody cytokine graft protein, and optionally a secretion signal.

33. The recombinant host cell of claim 32, which is a mammalian cell line.

34. The recombinant host cell of claim 33, wherein the mammalian cell line is a CHO cell line.

35. A pharmaceutical composition comprising the antibody cytokine graft protein of any one of claims 1-30, and a pharmaceutically acceptable carrier.

36. A method of treating cancer in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of the antibody cytokine graft protein of any one of claims 1-30 or the pharmaceutical composition of claim 35.

37. The method of claim 36, wherein the cancer is selected from the group consisting of: melanoma, lung cancer, colorectal cancer, prostate cancer, breast cancer, and lymphoma.

38. The method of claim 36 or 37, wherein the antibody cytokine graft protein or pharmaceutical composition is administered in combination with another therapeutic agent.

39. The method of claim 38, wherein the therapeutic agent is another antibody cytokine graft protein.

40. The method of claim 38, wherein the therapeutic agent is an immune checkpoint inhibitor.

41. The method of claim 40, wherein the immune checkpoint is selected from the group consisting of: PD-1, PD-L1, PD-L2, TIM3, CTLA-4, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGFR.

42. A method of expanding CD8T effector cells in a patient in need thereof, the method comprising administering to the patient the antibody cytokine graft protein of any one of claims 1-30 or the pharmaceutical composition of claim 35.

43. The method of claim 42, wherein the CD8T effector cells are expanded and Treg cells are not expanded.

44. The method of claim 42, wherein the CD8T effector cells are expanded and NK cells are not expanded.

45. The method of any one of claims 42-44, further comprising administering an immune checkpoint inhibitor.

46. The method of claim 45, wherein the immune checkpoint is selected from the group consisting of: PD-1, PD-L1, PD-L2, TIM3, CTLA-4, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGFR.

47. Use of an antibody cytokine graft protein in the treatment of cancer, the antibody cytokine graft protein comprising:

(i) a heavy chain variable region comprising (a) HCDR1 of SEQ ID NO:13, (b) HCDR2 of SEQ ID NO:14, (c) HCDR3 of SEQ ID NO:15, and a light chain variable region comprising: (d) LCDR1 of SEQ ID NO. 29, (e) LCDR2 of SEQ ID NO. 30, and (f) LCDR3 of SEQ ID NO. 31; or

(ii) A heavy chain variable region comprising (a) HCDR1 of SEQ ID NO:45, (b) HCDR2 of SEQ ID NO:46, (c) HCDR3 of SEQ ID NO: 47; and a light chain variable region comprising: (d) LCDR1 of SEQ ID NO:61, (e) LCDR2 of SEQ ID NO:62, and (f) LCDR3 of SEQ ID NO: 63.

48. The use of claim 47, wherein the antibody cytokine graft protein is administered in combination with another therapeutic agent.

49. The use of claim 48, wherein the therapeutic agent is an immune checkpoint inhibitor.

50. The use of claim 49, wherein the immune checkpoint is selected from the group consisting of: PD-1, PD-L1, PD-L2, TIM3, CTLA-4, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGFR.