MESOTHELIN CHIMERIC ANTIGEN RECEPTOR (CAR) AND ANTIBODY AGAINST PD-L1 INHIBITOR FOR COMBINED USE IN ANTICANCER THERAPY

• US 20190000944A1
• Filed: 12/21/2016
• Published: 01/03/2019
• Est. Priority Date: 12/22/2015
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute, President and Fellows of Harvard College (Harvard Corporation), Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Dana-Farber Cancer Institute, President and Fellows of Harvard College (Harvard Corporation), Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Brogdon, Jennifer, Chang, Hwai Wen, Engels, Boris, Freeman, Gordon James, Frey, Gerhard Johann, Mataraza, Jennifer Marie, Singh, Reshma, Sharpe, Arlene Helen
• Granted Patent: US 11,413,340 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/4636 Immune checkpoint inhibitors
• CPC Class Codes: A61K 39/464468 Mesothelin [MSLN]
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

Provided are compositions for use in methods for treating diseases associated with expression of mesothelin comprising administering a cell that expresses a chimeric antigen receptor (CAR) specific to mesothelin in combination with a PD-L1 inhibitor.

Claims

1. A method of treating a subject having a disease associated with mesothelin expression, comprising administering to the subject

i) a cell, e.g., a population of immune effector cells, comprising, e.g., expressing, a chimeric antigen receptor (CAR), wherein the CAR comprises a mesothelin binding domain, a transmembrane domain, and an intracellular signaling domain;

and

ii) a PD-L1 inhibitor, wherein the PD-L1 inhibitor is administered prior to administration of the cell comprising a CAR.

2. The method of claim 1, wherein the CAR-expressing cell and the PD-L1 inhibitor is administered for a treatment interval, and wherein the treatment interval comprises a single dose of the PD-L1 inhibitor and a single dose of the CAR-expressing cell.

3. The method of claim 2, wherein the treatment interval is initiated upon administration of the dose of the PD-L1 inhibitor and completed upon administration of the dose of the CAR-expressing cell.

4. The method of claim 2 or 3, wherein the treatment interval further comprises one or more, e.g., 1, 2, 3, 4, or 5 or more, subsequent doses of the PD-L1 inhibitor.

5. The method of any of claims 2-4, wherein the dose of the CAR-expressing cell is administered at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 2 weeks after the dose of PD-L1 inhibitor is administered.

6. The method of claim 5, wherein the dose of the CAR-expressing cell is administered 2 days after the dose of the PD-L1 inhibitor is administered.

7. The method of claim 1, wherein the CAR-expressing cell and the PD-L1 inhibitor is administered for a treatment interval, wherein the treatment interval comprises a first and second dose of the PD-L1 inhibitor and a dose of the CAR-expressing cell, and wherein the dose of the CAR-expressing cell is administered after administration of the first dose of the PD-L1 inhibitor but before the administration of the second dose of the PD-L1 inhibitor.

8. The method of claim 7, wherein the treatment interval is initiated upon administration of the first dose of the PD-L1 inhibitor and completed upon administration of the second dose of the PD-L1 inhibitor.

9. The method of claim 7 or 8, wherein the second dose of the PD-L1 inhibitor is administered at least 5 days, 7 days, 1 week, 2 weeks, or 3 weeks after administration of the first dose of the PD-L1 inhibitor.

10. The method of any of claims 7-9, wherein the dose of the CAR-expressing cell is administered at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 2 weeks after administration of the first dose of the PD-L1 inhibitor.

11. The method of any of claims 7-10, wherein the second dose of the PD-L1 inhibitor is administered at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 2 weeks after administration of the dose of the CAR-expressing cell.

12. The method of any of claims 2-11, wherein the treatment interval is repeated, e.g., one or more times, e.g., 1, 2, 3, 4, or 5 more times.

13. The method of any of claims 2-11, wherein the treatment interval is followed by one or more, e.g., 1, 2, 3, 4, or 5, subsequent treatment intervals.

14. The method of claim 13, wherein the one or more subsequent treatment interval is different from the first or previous treatment interval.

15. The method of claim 13 or 14, wherein the one or more subsequent treatment intervals is administered at least 1 day, e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 2 weeks, after the completion of the first or previous treatment interval.

16. The method of claims 2-15, wherein one or more subsequent doses, e.g., 1, 2, 3, 4, or 5 or more doses, of the PD-L1 inhibitor is administered after the completion of one or more treatment intervals.

17. The method of claim 16, wherein a dose of the PD-L1 inhibitor is administered every 5 days, 7 days, 2 weeks, 3 weeks, or 4 days after the completion of one or more treatment intervals.

18. The method of any of claims 2-6, wherein the treatment interval comprises a dose of CAR-expressing cells administered 2 days after the dose of the PD-L1 inhibitor is administered, and wherein the treatment interval is repeated twice, and wherein the treatment intervals are initiated 3 days after the completion of the previous treatment interval.

19. The method of claim 18, wherein one or more subsequent doses of the PD-L1 inhibitor is administered every 5 days, 7 days, 2 weeks, 3 weeks, or 4 weeks, after the second treatment interval.

20. The method of claim 1, wherein the subject is administered a single dose of a CAR-expressing cell and a single dose of a PD-L1 inhibitor.

21. The method of claim 20, wherein the single dose of the CAR-expressing cell is administered at least 2 days, e.g., 2, 3, 4, 5, 6, 7 days, or 2 weeks, after administration of the single dose of the PD-L1 inhibitor.

22. The method of claim 21, wherein one or more, e.g., 1, 2, 3, 4, or 5, subsequent doses of a CAR-expressing cell is administered to the subject after the initial dose of the CAR-expressing cell.

23. The method of claim 22, wherein the one or more subsequent doses of the CAR-expressing cell are administered at least 2 days, e.g., 2, 3, 4, 5, 6, 7 days or 2 weeks, after the previous dose of the CAR-expressing cell.

24. The method of claim 23, wherein the one or more subsequent doses of the CAR-expressing cell are administered at least 5 days after the previous dose of the CAR-expressing cell.

25. The method of any of claims 21-24, wherein one or more, e.g., 1, 2, 3, 4, or 5, subsequent doses of PD-L1 inhibitor are administered after administration of the single dose of the PD-L1 inhibitor.

26. The method of claim 25, wherein the one or more subsequent doses of the PD-L1 inhibitor are administered at least 5 days, 7 days, 2 weeks, 3 weeks or 4 weeks, after the previous dose of PD-L1 inhibitor.

27. The method of claim 25, wherein the one or more subsequent doses of the PD-L1 inhibitor are administered at least 1, 2, 3, 4, 5, 6, or 7 days, after a dose of the CAR-expressing cell, e.g., the initial dose of the CAR-expressing cell.

28. The method of claim 25, wherein one or more, e.g., 1, 2, 3, 4, or 5, doses of the PD-L1 inhibitor is administered prior to the first dose of the CAR-expressing cell.

29. The method of any of claims 20-28, wherein the administration of the one or more doses of the CAR-expressing cell and the one or more doses of PD-L1 inhibitor is repeated.

30. The method of any of the preceding claims, wherein the dose of CAR-expressing cells comprises at least about 1-3×

10⁷ to 1-3×

10⁸.

31. The method claim 30, wherein the dose of CAR-expressing cells is about 1-3×

10⁸ cells.

32. The method of claim 30, wherein the dose of CAR-expressing cells is about 1-3×

10⁸ cells.

33. The method of any of the preceding claims, wherein the dose of the PD-L1 inhibitor is between 1 and 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20 mg/kg, or about 1 to 5 mg/kg.

34. The method of claim 33, wherein the dose of the PD-L1 inhibitor is about 10 to 20 mg/kg.

35. The method of any of the preceding claims, wherein the PD-L1 inhibitor comprises an antibody molecule, a small molecule, a polypeptide, e.g., a fusion protein, or an inhibitory nucleic acid, e.g., a siRNA or shRNA.

36. The method of any of the preceding claims, wherein the PD-L1 inhibitor is characterized by one or more of the following:

i) inhibits or reduces PD-L1 expression, e.g., transcription or translation of PD-L1;

ii) inhibits or reduces PD-L1 activity, e.g., inhibits or reduces binding of PD-L1 to its cognate receptor, e.g., PD-1;

or

iii) binds to PD-L1 or its receptor, e.g., PD-1.

37. The method of any of the preceding claims, wherein the PD-L1 inhibitor is an antibody molecule.

38. The method of of the preceding claims, wherein the PD-L1 inhibitor is selected from the group consisting of YW243.55.S70, MPDL3280A (atezolizumab), MEDI-4736, MSB-0010718C (avelumab), MDX-1105, and any anti-PD-L1 antibody molecule provided in Table 6.

39. The method of any of the preceding claims, wherein the PD-L1 inhibitor comprises an anti-PD-L1 antibody molecule comprising

i) a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), and a heavy chain complementary determining region 3 (HC CDR3) of any PD-L1 antibody molecule amino acid sequence listed in Table 6;

and

ii) a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), and a light chain complementary determining region 3 (LC CDR3) of any PD-L1 antibody molecule amino acid sequence listed in Table 6.

40. The method of claim 39 wherein the anti-PD-L1 antibody molecule thereof comprises

i) a) a HC CDR1 amino acid sequence chosen from SEQ ID NO:

287, 290, or 195, a HC CDR2 amino acid sequence of SEQ ID NO;

288, and a HC CDR3 amino acid sequence of SEQ ID NO;

289; and

b) a LC CDR1 amino acid sequence of SEQ ID NO;

295, a LC CDR2 amino acid sequence of SEQ ID NO;

296, and a LC CDR3 amino acid sequence of SEQ ID NO;

297;

or

ii) a) a HC CDR1 amino acid sequence chosen from SEQ ID NO;

287, 290, or 195, a HC CDR2 amino acid sequence of SEQ ID NO;

291, and a HC CDR3 amino acid sequence of SEQ ID NO;

292; and

b) a LC CDR1 amino acid sequence of SEQ ID NO;

298, a LC CDR2 amino acid sequence of SEQ ID NO;

299, and a LC CDR3 amino acid sequence of SEQ ID NO;

300.

41. The method of claim 39 or 40, wherein the anti-PD-L1 antibody molecule comprises a heavy chain variable region comprising:

i) the amino acid sequence of any heavy chain variable region listed in Table 6, e.g., SEQ ID NOs;

304, 316, 324, 332, 336, 340, 348, 356, or 364;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to the amino acid sequence of any heavy chain variable region provided in Table 6, e.g., SEQ ID NOs;

304, 316, 324, 332, 336, 340, 348, 356, or 364;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any heavy chain variable region provided in Table 6, e.g., SEQ ID NOs;

304, 316, 324, 332, 336, 340, 348, 356, or 364.

42. The method of any of claims 39-41, wherein the anti-PD-L1 antibody molecule comprises a heavy chain comprising:

i) the amino acid sequence of any heavy chain listed in Table 6, e.g., SEQ ID NOs;

306, 318, 326, 334, 338, 342, 350, 358, 366, 393, 377, or 382;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any heavy chain listed in Table 6, e.g., SEQ ID NOs;

306, 318, 326, 334, 338, 342, 350, 358, 366, 393, 377, or 382;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any heavy chain listed in Table 6, e.g., SEQ ID NOs;

306, 318, 326, 334, 338, 342, 350, 358, 366, 393, 377, or 382.

43. The method of any of claims 39-42, wherein the anti-PD-L1 antibody molecule comprises a light chain variable region comprising:

i) the amino acid sequence of any light chain variable region listed in Table 6, e.g., SEQ ID NOs;

308, 312, 320, 328, 344, 352, 360, 368, or 372;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to the amino acid sequence of any light chain variable region provided in Table 6, e.g., SEQ ID NOs;

308, 312, 320, 328, 344, 352, 360, 368, or 372;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any light chain variable region provided in Table 6, e.g., SEQ ID NOs;

308, 312, 320, 328, 344, 352, 360, 368, or 372.

44. The method of any of claims 39-43, wherein the anti-PD-L1 antibody molecule comprises a light chain comprising:

i) the amino acid sequence of any light chain listed in Table 6, e.g., SEQ ID NOs;

310, 314, 322, 330, 346, 354, 362, 370, or 374;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any light chain listed in Table 6, e.g., SEQ ID NOs;

310, 314, 322, 330, 346, 354, 362, 370, or 374;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence to any any light chain listed in Table 6, e.g., SEQ ID NOs;

310, 314, 322, 330, 346, 354, 362, 370, or 374.

45. The method of any of claims 39-44, wherein the anti-PD-L1 antibody molecule comprises:

i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

304 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

308;

ii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

304 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

312;

iii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

304 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

372.

iv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

316 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

320;

v) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

316 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

352;

vi) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

324 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

328;

vii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

324 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

360;

viii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

332 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

328;

ix) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

336 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

328;

x) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

336 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

308;

xi) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

336 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

372;

xii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

340 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

344;

xiii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

340 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

372;

xiv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

348 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

352;

xv) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

348 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

386;

xvi) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

356 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

352;

or

xvii) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO;

78 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO;

368.

46. The method of any of claims 39-45, wherein the anti-PD-L1 antibody molecule comprises:

i) a heavy chain comprising the amino acid sequence of SEQ ID NO;

306 and a light chain comprising the amino acid sequence of SEQ ID NO;

310;

ii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

306 and a light chain comprising the amino acid sequence of SEQ ID NO;

214;

iii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

306 and a light chain comprising the amino acid sequence of SEQ ID NO;

374;

iv) a heavy chain comprising the amino acid sequence of SEQ ID NO;

318 and a light chain comprising the amino acid sequence of SEQ ID NO;

322;

v) a heavy chain comprising the amino acid sequence of SEQ ID NO;

318 and a light chain comprising the amino acid sequence of SEQ ID NO;

354;

vi) a heavy chain comprising the amino acid sequence of SEQ ID NO;

326 and a light chain comprising the amino acid sequence of SEQ ID NO;

330;

vii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

326 and a light chain comprising the amino acid sequence of SEQ ID NO;

362;

viii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

334 and a light chain comprising the amino acid sequence of SEQ ID NO;

330;

ix) a heavy chain comprising the amino acid sequence of SEQ ID NO;

338 and a light chain comprising the amino acid sequence of SEQ ID NO;

330;

x) a heavy chain comprising the amino acid sequence of SEQ ID NO;

338 and a light chain comprising the amino acid sequence of SEQ ID NO;

310;

xi) a heavy chain comprising the amino acid sequence of SEQ ID NO;

338 and a light chain comprising the amino acid sequence of SEQ ID NO;

374;

xii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

342 and a light chain comprising the amino acid sequence of SEQ ID NO;

346;

xiii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

342 and a light chain comprising the amino acid sequence of SEQ ID NO;

374;

xiv) a heavy chain comprising the amino acid sequence of SEQ ID NO;

350 and a light chain comprising the amino acid sequence of SEQ ID NO;

354;

xv) a heavy chain comprising the amino acid sequence of SEQ ID NO;

350 and a light chain comprising the amino acid sequence of SEQ ID NO;

374;

xvi) a heavy chain comprising the amino acid sequence of SEQ ID NO;

358 and a light chain comprising the amino acid sequence of SEQ ID NO;

354;

xvii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

366 and a light chain comprising the amino acid sequence of SEQ ID NO;

370;

xviii) a heavy chain comprising the amino acid sequence of SEQ ID NO;

393 and a light chain comprising the amino acid sequence of SEQ ID NO;

322;

xix) a heavy chain comprising the amino acid sequence of SEQ ID NO;

91 and a light chain comprising the amino acid sequence of SEQ ID NO;

330;

or

xx) a heavy chain comprising the amino acid sequence of SEQ ID NO;

382 and a light chain comprising the amino acid sequence of SEQ ID NO;

354.

47. The method of any of the preceding claims, wherein the mesothelin binding domain comprises a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), and a heavy chain complementary determining region 3 (HC CDR3) of any mesothelin heavy chain binding domain amino acid sequence listed in Table 2;

and a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), and a light chain complementary determining region 3 (LC CDR3) of any mesothelin light chain binding domain amino acid sequence listed in Table 2.

48. The method of claim 47, wherein the mesothelin binding domain comprises a HC CDR1, a HC CDR2, and a HC CDR3 according to the HC CDR amino acid sequences in Table 4, and a LC CDR1, a LC CDR2, and a LC CDR3 according to the LC CDR amino acid sequences in Table 5.

49. The method of any of the preceding claims, wherein the mesothelin binding domain comprises:

i) the amino acid sequence of any heavy chain variable region of a mesothelin binding domain listed in Table 2;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to the amino acid sequence of any heavy chain variable region of a mesothelin binding domain provided in Table 2;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any heavy chain variable region of a mesothelin binding domain provided in Table 2.

50. The method of any of the preceding claims, wherein the mesothelin binding domain comprises:

i) the amino acid sequence of any heavy chain of a mesothelin binding domain provided in Table 2;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any heavy chain of a mesothelin binding domain provided in Table 2;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence to any heavy chain of a mesothelin binding domain provided in Table 2.

51. The method of any of the preceding claims, wherein the mesothelin binding domain comprises:

i) the amino acid sequence of any light chain variable region of a mesothelin binding domain provided in Table 2;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to the amino acid sequence of any light chain variable region of a mesothelin binding domain provided in Table 2;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any light chain variable region of a mesothelin binding domain provided in Table 2.

52. The method of any of the preceding claims, wherein the mesothelin binding domain comprises:

i) the amino acid sequence of any light chain of a mesothelin binding domain provided in Table 2;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any light chain of a mesothelin binding domain provided in Table 2;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence to any light chain of a mesothelin binding domain provided in Table 2.

53. The method of any of the preceding claims, wherein the mesothelin binding domain comprises the amino acid sequence of any heavy chain variable region listed in Table 2, and the amino acid sequence of any light chain variable region listed in Table 2.

54. The method of any of the preceding claims, wherein the mesothelin binding domain comprises:

i) the amino acid sequence selected from the group consisting of SEQ ID NO;

43, SEQ ID NO;

49, SEQ ID NO;

275, SEQ ID NO;

39, SEQ ID NO;

40, SEQ ID NO;

41, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

45, SEQ ID NO;

46, SEQ ID NO;

47, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

51, SEQ ID NO;

52, SEQ ID NO;

53, SEQ ID NO;

54, SEQ ID NO;

55, SEQ ID NO;

56, SEQ ID NO;

57, SEQ ID NO;

58, SEQ ID NO;

59, SEQ ID NO;

60, SEQ ID NO;

61, or SEQ ID NO;

62;

ii) the amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NO;

43, SEQ ID NO;

49, SEQ ID NO;

275, SEQ ID NO;

39, SEQ ID NO;

40, SEQ ID NO;

41, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

45, SEQ ID NO;

46, SEQ ID NO;

47, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

51, SEQ ID NO;

52, SEQ ID NO;

53, SEQ ID NO;

54, SEQ ID NO;

55, SEQ ID NO;

56, SEQ ID NO;

57, SEQ ID NO;

58, SEQ ID NO;

59, SEQ ID NO;

60, SEQ ID NO;

61, or SEQ ID NO;

62;

or

iii) an amino acid sequence with 95-99% identity to the amino acid sequence to any of SEQ ID NO;

43, SEQ ID NO;

49, SEQ ID NO;

275, SEQ ID NO;

39, SEQ ID NO;

40, SEQ ID NO;

41, SEQ ID NO;

42, SEQ ID NO;

44, SEQ ID NO;

45, SEQ ID NO;

46, SEQ ID NO;

47, SEQ ID NO;

48, SEQ ID NO;

50, SEQ ID NO;

51, SEQ ID NO;

52, SEQ ID NO;

53, SEQ ID NO;

54, SEQ ID NO;

55, SEQ ID NO;

56, SEQ ID NO;

57, SEQ ID NO;

58, SEQ ID NO;

59, SEQ ID NO;

60, SEQ ID NO;

61, or SEQ ID NO;

62.

55. The method of any of the preceding claims, wherein the transmembrane domain comprises a transmembrane domain from a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.

56. The method of any of the preceding claims, wherein the transmembrane domain comprises

(i) the amino acid sequence of SEQ ID NO:

6,

(ii) an amino acid sequence comprises at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

6, or

(iii) a sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

6.

57. The method of any of the preceding claims, wherein the mesothelin binding domain is connected to the transmembrane domain by a hinge region.

58. The method of any of the preceding claims, wherein the hinge region comprises SEQ ID NO:

2, or a sequence with 95-99% identity thereof.

59. The method of any of the preceding claims, wherein the intracellular signaling domain comprises a costimulatory signaling domain comprising a functional signaling domain obtained from a protein selected from the group consisting of a MHC class I molecule, a TNF receptor protein, an Immunoglobulin-like protein, a cytokine receptor, an integrin, a signaling lymphocytic activation molecule (SLAM protein), an activating NK cell receptor, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

60. The method of claim 59, wherein the costimulatory domain comprises the amino acid sequence of SEQ ID NO:

7, or an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

7, or an amino acid sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

7.

61. The method of any of the preceding claims, wherein the intracellular signaling domain comprises a functional signaling domain of 4-1BB and/or a functional signaling domain of CD3 zeta.

62. The method of any of the preceding claims, wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:

7 and/or the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10;

or an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

7 and/or the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10;

or an amino acid sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

7 and/or the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10.

63. The method of any of the preceding claims, wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:

7 and the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10, wherein the amino acid sequences comprising the intracellular signaling domain are expressed in the same frame and as a single polypeptide chain.

64. The method of any of the preceding claims, wherein the CAR further comprises a leader sequence comprising the amino acid sequence of SEQ ID NO:

1.

65. The method of any of the preceding claims, wherein the CAR comprises:

(i) the amino acid sequence of any of SEQ ID NO;

67;

SEQ ID NO;

73, SEQ ID NO;

278, SEQ ID NO;

63, SEQ ID NO;

64, SEQ ID NO;

65, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

69, SEQ ID NO;

70, SEQ ID NO;

71, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

75, SEQ ID NO;

76, SEQ ID NO;

77, SEQ ID NO;

78, SEQ ID NO;

79, SEQ ID NO;

80, SEQ ID NO;

81, SEQ ID NO;

82, SEQ ID NO;

83, SEQ ID NO;

84, SEQ ID NO;

85, or SEQ ID NO;

86;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NO;

67, SEQ ID NO;

73, SEQ ID NO;

278, SEQ ID NO;

63, SEQ ID NO;

64, SEQ ID NO;

65, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

69, SEQ ID NO;

70, SEQ ID NO;

71, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

75, SEQ ID NO;

76, SEQ ID NO;

77, SEQ ID NO;

78, SEQ ID NO;

79, SEQ ID NO;

80, SEQ ID NO;

81, SEQ ID NO;

82, SEQ ID NO;

83, SEQ ID NO;

84, SEQ ID NO;

85, or SEQ ID NO;

86;

or

(iii) an amino acid sequence with 95-99% identity to any of SEQ ID NO;

67;

SEQ ID NO;

73, SEQ ID NO;

278, SEQ ID NO;

63, SEQ ID NO;

64, SEQ ID NO;

65, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

69, SEQ ID NO;

70, SEQ ID NO;

71, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

75, SEQ ID NO;

76, SEQ ID NO;

77, SEQ ID NO;

78, SEQ ID NO;

79, SEQ ID NO;

80, SEQ ID NO;

81, SEQ ID NO;

82, SEQ ID NO;

83, SEQ ID NO;

84, SEQ ID NO;

85, or SEQ ID NO;

86.

66. The method of any of the preceding claims, wherein the cell comprising a CAR comprises a nucleic acid encoding the CAR.

67. The method of claim 66, wherein the nucleic acid encoding the CAR is a lentiviral vector.

68. The method of claim 66 or 67, wherein the nucleic acid encoding the CAR is introduced into the cells by lentiviral transduction.

69. The method of any of claim 66, wherein the nucleic acid encoding the CAR is an RNA, e.g., an in vitro transcribed RNA.

70. The method of claim 69, wherein the nucleic acid encoding the CAR is introduced into the cells by electroporation.

71. The method of any of the preceding claims, wherein the cell is a T cell or an NK cell.

72. The method of claim 71, wherein the T cell is an autologous or allogeneic T cell.

73. The method of any of the preceding claims, further comprising administering an additional therapeutic agent that treats the disease associated with mesothelin expression, e.g., an anti-cancer agent.

74. The method of any of the preceding claims, wherein the disease associated with mesothelin expression is a cancer.

75. The method of claim 74, wherein the cancer is chosen from one or more of mesothelioma, malignant pleural mesothelioma, non-small cell lung cancer, small cell lung cancer, squamous cell lung cancer, or large cell lung cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, pancreatic metatstatic, ovarian cancer, or colorectal cancer and bladder cancer, or a metastasis thereof.

76. The method of any of the preceding claims, wherein the subject is a mammal, e.g., a human.

77. The method of any of the preceding claims, wherein the subject expresses PD-L1 and/or PD-L2.

78. The method of claim 74, wherein a cancer cell or a cell in close proximity to a cancer cell in the subject expresses PD-L1 and/or PD-L2.

79. The method of any of the preceding claims, wherein the cell expressing a CAR expresses PD-1 and/or PD-L1.

80. A combination comprising

a cell, e.g., a population of immune effector cells, comprising a CAR, wherein the CAR comprises a mesothelin binding domain, a transmembrane domain, and an intracellular signaling domain;

and

a PD-L1 inhibitor chosen from Table 6;

for use in treating a disease associated with mesothelin expression, e.g., a cancer, in a subject.

81. A composition (e.g., one or more compositions or dosage forms), comprising

a cell, e.g., a population of immune effector cells, comprising a CAR, wherein the CAR comprises a mesothelin binding domain, a transmembrane domain, and an intracellular signaling domain, and

a PD-L1 inhibitor chosen from Table 6.