Processing

Please wait...

Settings

Settings

Goto Application

1. WO2018129332 - EXPANSION OF TUMOR INFILTRATING LYMPHOCYTES (TILS) WITH TUMOR NECROSIS FACTOR RECEPTOR SUPERFAMILY (TNFRSF) AGONISTS AND THERAPEUTIC COMBINATIONS OF TILS AND TNFRSF AGONISTS

Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

[ EN ]

CLAIMS

We claim:

1. A method of treating a cancer with a population of tumor infiltrating lymphocytes (TILs) comprising the steps of:

(a) resecting a tumor from a patient;

(b) obtaining a first population of TILs from the tumor;

(c) performing an initial expansion of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the second population of TILs is at least 5-fold greater in number than the first population of TILs, wherein the first cell culture medium comprises IL-2 and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and wherein the initial expansion is performed over a period of 21 days or less;

(d) performing a rapid expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs, wherein the third population of TILs is at least 50-fold greater in number than the second population of TILs after 7 days from the start of the rapid expansion; wherein the second cell culture medium comprises IL-2, OKT-3 (anti- CD3) antibody, peripheral blood mononuclear cells (PBMCs), and optionally the TNFRSF agonist and a second TNFRSF agonist, and wherein the rapid expansion is performed over a period of 14 days or less;

(e) harvesting the third population of TILs; and

(f) administering a therapeutically effective portion of the third population of TILs to the patient.

2. The method of Claim 1, wherein the TNFRSF agonist is selected from the group consisting of a 4- IBB agonist, an OX40 agonist, a CD27 agonist, a GITR agonist, a HVEM agonist, a CD95 agonist, and combinations thereof.

3. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a 4- IBB agonist, and the 4- IBB agonist is selected from the group consisting of urelumab, utomilumab, EU- 101, a fusion protein, and fragments, derivatives, variants, biosimilars, and combinations thereof.

4. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is an OX40 agonist, or fragments, derivatives, variants, biosimilars, and combinations thereof.

5. The method of Claim 4, wherein the 4- IBB agonist fusion protein comprises (i) a first

soluble 4- IBB binding domain, (ii) a first peptide linker, (iii) a second soluble 4- IBB binding domain, (iv) a second peptide linker, and (v) a third soluble 4- IBB binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

6. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a OX40 agonist, and the OX40 agonist is selected from the group consisting of tavolixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM-18455, and fragments, derivatives, variants, biosimilars, and combinations thereof.

7. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is an OX40 agonist, and the OX40 agonist is an OX40 agonist fusion protein.

8. The method of Claim 7, wherein the OX40 agonist fusion protein comprises (i) a first soluble OX40 binding domain, (ii) a first peptide linker, (iii) a second soluble OX40 binding domain, (iv) a second peptide linker, and (v) a third soluble OX40 binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

9. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a CD27 agonist, and the CD27 agonist is varlilumab, or a fragment, derivative, variant, or biosimilar thereof.

10. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a CD27 agonist, and wherein the CD27 agonist is an CD27 agonist fusion protein.

11. The method of Claim 10, wherein the CD27 agonist fusion protein comprises (i) a first

soluble CD27 binding domain, (ii) a first peptide linker, (iii) a second soluble CD27 binding domain, (iv) a second peptide linker, and (v) a third soluble CD27 binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the

fusion protein is a dimeric structure according to structure I-A or structure I-B.

12. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a GITR agonist, and the GITR agonist is selected from the group consisting of TRX518, 6C8, 36E5, 3D6, 61G6, 6H6, 61F6, 1D8, 17F10, 35D8, 49A1, 9E5, 31H6, 2155, 698, 706, 827, 1649, 1718, 1D7, 33C9, 33F6, 34G4, 35B 10, 41E11, 41G5, 42A11, 44C1, 45A8, 46E11, 48H12, 48H7, 49D9, 49E2, 48 A9, 5H7, 7A10, 9H6, and fragments, derivatives, variants, biosimilars, and combinations thereof.

13. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is an GITR agonist, and the GITR agonist is a GITR agonist fusion protein.

14. The method of Claim 13, wherein the GITR agonist fusion protein comprises (i) a first

soluble GITR binding domain, (ii) a first peptide linker, (iii) a second soluble GITR binding domain, (iv) a second peptide linker, and (v) a third soluble GITR binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

15. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a HVEM agonist.

16. The method of Claim 15, and the HVEM agonist is a HVEM agonist fusion protein.

17. The method of Claim 16, wherein the HVEM agonist fusion protein comprises (i) a first soluble HVEM binding domain, (ii) a first peptide linker, (iii) a second soluble HVEM binding domain, (iv) a second peptide linker, and (v) a third soluble HVEM binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

18. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a CD95 agonist.

19. The method of any one of Claims 1 to 2, wherein the TNFRSF agonist is a CD95 agonist, and the CD95 agonist is a CD95 agonist fusion protein.

20. The method of Claim 19, wherein the CD95 agonist is a fusion protein comprising (i) a first soluble CD95 binding domain, (ii) a first peptide linker, (iii) a second soluble CD95 binding domain, (iv) a second peptide linker, and (v) a third soluble CD95 binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

21. The method of any one of Claims 1 to 20, further comprising the step of treating the patient with the TNFRSF agonist starting on the day after administration of the third population of TILs to the patient, wherein the TNFRSF agonist is administered intravenously at a dose of between 0.1 mg/kg and 50 mg/kg every four weeks for up to eight cycles.

22. The method of any one of Claims 1 to 21, further comprising the step of treating the patient with the TNFRSF agonist prior to the step of resecting of a tumor from the patient, wherein the TNFRSF agonist is administered intravenously at a dose of between 0.1 mg/kg and 50 mg/kg every four weeks for up to eight cycles.

23. The method of any one of Claims 1 to 22, wherein the TNFRSF agonist is selected from the group consisting of urelumab, utomilumab, EU-101, tavolixizumab, Creative Biolabs MOM- 18455, and fragments, derivatives, variants, biosimilars, and combinations thereof.

24. The method of any one of Claims 1 to 23, wherein the first cell culture medium comprises a second TNFRSF agonist.

25. The method of any one of Claims 1 to 23, wherein the TNFRSF agonist is a 4-1BB agonist, and the second TNFRSF agonist is an OX40 agonist.

26. The method of any one of Claims 1 to 25, wherein the TNFRSF agonist is added to the first cell culture medium during the initial expansion at an interval selected from the group consisting of every day, every two days, every three days, every four days, every five days, every six days, every seven days, and every two weeks.

27. The method of any one of Claims 1 to 26, wherein the TNFRSF agonist is added to the

second cell culture medium during the rapid expansion at an interval selected from the group consisting of every day, every two days, every three days, every four days, every five days, every six days, every seven days, and every two weeks.

28. The method of any one of Claims 24 to 27, wherein the TNFRSF agonist is added at a

concentration sufficient to achieve a concentration in the cell culture medium of between 0.1 μg/mL and 100 μg/mL.

29. The method of Claim 28, wherein the TNFRSF agonist is added at a concentration sufficient to achieve a concentration in the cell culture medium of between 20 μg/mL and 40 μg/mL.

30. The method of any one of Claims 1 to 30, wherein IL-2 is present at an initial concentration of about 10 to about 6000 R7/mL in the first cell culture medium.

31. The method of Claim 31, wherein IL-2 is present at an initial concentration of about 3000 IU/mL in the first cell culture medium.

32. The method of Claim 31, wherein IL-2 is present at an initial concentration of about 800 to about 1100 IU/mL in the first cell culture medium.

33. The method of Claim 33, wherein IL-2 is present at an initial concentration of about 1000 IU/mL in the first cell culture medium.

34. The method of any one of Claims 1 to 30, wherein IL-2 is present at an initial concentration of about 10 to about 6000 R7/mL in the second cell culture medium.

35. The method of Claim 35, wherein IL-2 is present at an initial concentration of about 3000 IU/mL in the second cell culture medium.

36. The method of Claim 35, wherein IL-2 is present at an initial concentration of about 800 to about 1100 IU/mL in the second cell culture medium.

37. The method of Claim 35, wherein IL-2 is present at an initial concentration of about 1000 IU/mL in the second cell culture medium.

38. The method of any one of Claims 1 to 37, wherein IL-15 is present in the first cell culture medium.

39. The method of Claim 38, wherein IL-15 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the first cell culture medium.

40. The method of any one of Claims 1 to 39, wherein IL-15 is present in the second cell culture medium.

41. The method of Claim 40, wherein IL-15 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the second cell culture medium.

42. The method of any one of Claims 1 to 41, wherein IL-21 is present in the first cell culture medium.

43. The method of Claim 42, wherein IL-21 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the first cell culture medium.

44. The method of any one of Claims 1 to 43, wherein IL-21 is present in the second cell culture medium.

45. The method of Claim 44, wherein IL-21 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the second cell culture medium.

46. The method of any one of Claims 1 to 45, wherein OKT-3 antibody is present at an initial concentration of about 10 ng/mL to about 60 ng/mL in the second cell culture medium.

47. The method of Claim 46, wherein OKT-3 antibody is present at an initial concentration of about 30 ng/mL in the second cell culture medium.

48. The method of any one of Claims 1 to 47, wherein the initial expansion is performed using a gas permeable container.

49. The method of any one of Claims 1 to 48, wherein the rapid expansion is performed using a gas permeable container.

50. The method of any one of Claims 1 to 49, further comprising the step of treating the patient with a non-myeloablative lymphodepletion regimen prior to administering the third population of TILs to the patient.

51. The method of Claim 50, wherein the non-myeloablative lymphodepletion regimen

comprises the steps of administration of cyclophosphamide at a dose of 60 mg/m2/day for two days followed by administration of fludarabine at a dose of 25 mg/m2/day for five days.

52. The method of any one of Claims 1 to 51, further comprising the step of treating the patient with a decrescendo IL-2 regimen starting on the day after administration of the third population of TILs to the patient, wherein the decrescendo IL-2 regimen comprises aldesleukin administered intravenously at a dose of 18,000,000 IU/m2 on day 1, 9,000,000 IU/m2 on day 2, and 4,500,000 IU/m2 on days 3 and 4.

53. The method of any one of Claims 1 to 51, further comprising the step of treating the patient with pegylated IL-2 after administration of the third population of TILs to the patient at a dose of 0.10 mg/day to 50 mg/day.

54. The method of any one of Claims 1 to 51, further comprising the step of treating the patient with a high-dose IL-2 regimen starting on the day after administration of the third population of TILs to the patient.

55. The method of Claim 54, wherein the high-dose IL-2 regimen comprises 600,000 or 720,000 IU/kg of aldesleukin, or a biosimilar or variant thereof, administered as a 15-minute bolus intravenous infusion every eight hours until tolerance.

56. The method of any one of Claims 1 to 55, wherein the cancer is selected from the group consisting of melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer, renal cell carcinoma, acute myeloid leukemia, colorectal cancer, cholangiocarcinoma, and sarcoma.

57. The method of any one of Claims 1 to 56, wherein the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC), triple negative breast cancer, double- refractory melanoma, and uveal (ocular) melanoma.

58. The method of any one of Claims 1 to 57, further comprising the step of treating the patient with a PD-1 inhibitor or PD-Ll inhibitor prior to resecting the tumor from the patient.

59. The method of Claim 58, wherein the PD-1 inhibitor or PD-Ll inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

60. The method of any one of Claims 1 to 59, further comprising the step of treating the patient with a PD-1 inhibitor or PD-Ll inhibitor after resecting the tumor from the patient.

61. The method of Claim 60, wherein the PD-1 inhibitor or PD-Ll inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

62. The method of any one of Claims 1 to 61, further comprising the step of treating the patient with a PD-1 inhibitor or PD-Ll inhibitor after administering the third population of TILs to the patient.

63. The method of Claim 62, wherein the PD-1 inhibitor or PD-Ll inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

64. The method of any one of Claims 1 to 63, wherein the first cell culture medium further

comprises IL-4, IL-7, or a combination thereof.

65. The method of any one of Claims 1 to 64, wherein the second cell culture medium further comprises IL-4, IL-7, or a combination thereof.

66. The method of any one of Claims 1 to 65, wherein the initial expansion is performed over a period of 11 days or less.

67. The method of any one of Claims 1 to 65, wherein the rapid expansion is performed over a period of 11 days or less.

68. A process for the preparation of a population of tumor infiltrating lymphocytes (TILs)

comprising the steps of:

(b) obtaining a first population of TILs;

(c) performing an initial expansion of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the second population of TILs is at least 5-fold greater in number than the first population of TILs, wherein the first cell culture medium comprises IL-2 and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and wherein the initial expansion is performed over a period of 21 days or less;

(d) performing a rapid expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs, wherein the third population of TILs is at least 50-fold greater in number than the second population of TILs after 7 days from the start of the rapid expansion; wherein the second cell culture medium comprises IL-2, OKT-3 (anti- CD3 antibody), peripheral blood mononuclear cells (PBMCs), and optionally the TNFRSF agonist, and wherein the rapid expansion is performed over a period of 14 days or less; and

(e) harvesting the third population of TILs.

69. The process according to claim 68 wherein the first population of TILs is obtained from a tumor which tumor has been resected from a patient.

70. The process according to any one of claims 68 to 69, wherein the TNFRSF agonist is selected from the group consisting of a 4-lBB agonist, an OX40 agonist, a CD27 agonist, a GITR agonist, a HVEM agonist, a CD95 agonist, and combinations thereof.

71. The process according to any one of claims 68 to 70, wherein the TNFRSF agonist is a 4- 1BB agonist.

72. The process according to any one of claims 68 to 71, the TNFRSF agonist is a 4- IBB

agonist, and the 4-lBB agonist is selected from the group consisting of urelumab, utomilumab, EU-101, and fragments, derivatives, variants, biosimilars, and combinations thereof.

73. The process according to any one of claims 68 to 72, wherein the TNFRSF agonist is a 4- 1BB agonist, and the 4- IBB agonist is a 4- IBB agonist fusion protein.

74. The process according to any one of claims 68 to 73, wherein the TNFRSF agonist is a 4- 1BB agonist fusion protein, and the 4-lBB agonist fusion protein comprises (i) a first soluble 4-lBB binding domain, (ii) a first peptide linker, (iii) a second soluble 4-lBB binding domain, (iv) a second peptide linker, and (v) a third soluble 4- IBB binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

75. The process according to any one of claims 68 to 74, wherein the TNFRSF agonist is a OX40 agonist.

76. The process according to any one of claims 68 to 75, wherein the TNFRSF agonist is a OX40 agonist, and the OX40 agonist is selected from the group consisting of tavolixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM- 18455, and fragments, derivatives, variants, biosimilars, and combinations thereof.

77. The process according to any one of claims 68 to 76, wherein the TNFRSF agonist is an OX40 agonist, and the OX40 agonist is an OX40 agonist fusion protein.

78. The process according to any one of claims 68 to 77, wherein the TNFRSF agonist is an OX40 agonist fusion protein, and the OX40 agonist fusion protein comprises (i) a first soluble OX40 binding domain, (ii) a first peptide linker, (iii) a second soluble OX40 binding domain, (iv) a second peptide linker, and (v) a third soluble OX40 binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

79. The process according to any one of claims 68 to 78, wherein the TNFRSF agonist is a CD27 agonist.

80. The process according to any one of claims 68 to 79, wherein the TNFRSF agonist is a CD27 agonist, and the CD27 agonist is varlilumab, or a fragment, derivative, variant, or biosimilar thereof.

81. The process according to any one of claims 68 to 80, wherein the TNFRSF agonist is a CD27 agonist, and wherein the CD27 agonist is an CD27 agonist fusion protein.

82. The process according to any one of claims 68 to 81, wherein the TNFRSF agonist is a CD27 agonist, and the CD27 agonist fusion protein comprises (i) a first soluble CD27 binding domain, (ii) a first peptide linker, (iii) a second soluble CD27 binding domain, (iv) a second peptide linker, and (v) a third soluble CD27 binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

83. The process according to any one of claims 68 to 82, wherein the TNFRSF agonist is a GITR agonist.

84. The process according to any one of claims 68 to 83, wherein the TNFRSF agonist is a GITR agonist, and the GITR agonist is selected from the group consisting of TRX518, 6C8, 36E5, 3D6, 61G6, 6H6, 61F6, 1D8, 17F10, 35D8, 49A1, 9E5, 31H6, 2155, 698, 706, 827, 1649, 1718, 1D7, 33C9, 33F6, 34G4, 35B 10, 41E11, 41G5, 42A11, 44C1, 45A8, 46E11, 48H12, 48H7, 49D9, 49E2, 48 A9, 5H7, 7A10, 9H6, and fragments, derivatives, variants, biosimilars, and combinations thereof.

85. The process according to any one of claims 68 to 84, wherein the TNFRSF agonist is an

GITR agonist and the GITR agonist is a GITR agonist fusion protein.

86. The process according to any one of claims 68 to 85, wherein the TNFRSF agonist is a GITR agonist fusion protein, and the GITR agonist fusion protein comprises (i) a first soluble GITR binding domain, (ii) a first peptide linker, (iii) a second soluble GITR binding domain, (iv) a second peptide linker, and (v) a third soluble GITR binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I- A or structure I-B.

87. The process according to any one of claims 68 to 86, wherein the TNFRSF agonist is a

HVEM agonist.

88. The process according to any one of claims 68 to 87, wherein the TNFRSF agonist is an HVEM agonist, and the HVEM agonist is a HVEM agonist fusion protein.

89. The process according to any one of claims 68 to 88, wherein the TNFRSF agonist is a

HVEM agonist fusion protein, and wherein the HVEM agonist fusion protein comprises (i) a first soluble HVEM binding domain, (ii) a first peptide linker, (iii) a second soluble HVEM binding domain, (iv) a second peptide linker, and (v) a third soluble HVEM binding domain, further comprising an additional domain at the N-terminal and/or C-terminal end, and wherein the additional domain comprises a Fc fragment domain and hinge domain, and wherein the fusion protein is a dimeric structure according to structure I-A or structure I-B.

90. The process according to any one of claims 68 to 89, wherein the TNFRSF agonist is

selected from the group consisting of urelumab, utomilumab, EU-101, tavolixizumab, Creative Biolabs MOM-18455, and fragments, derivatives, variants, biosimilars, and combinations thereof.

91. The process according to any one of claims 68 to 90, wherein the first cell culture medium comprises a second TNFRSF agonist.

92. The process according to any one of claims 68 to 91, wherein the TNFRSF agonist is added to the first cell culture medium during the initial expansion at an interval selected from the group consisting of every day, every two days, every three days, every four days, every five days, every six days, every seven days, and every two weeks.

93. The process according to any one of claims 68 to 92, wherein the TNFRSF agonist is added to the second cell culture medium during the rapid expansion at an interval selected from the group consisting of every day, every two days, every three days, every four days, every five days, every six days, every seven days, and every two weeks.

94. The process according to any one of claims 68 to 93, wherein the TNFRSF agonist is added at a concentration sufficient to achieve a concentration in the cell culture medium of between 0.1 μg/mL and 100 μg/mL.

95. The process according to any one of claims 68 to 94, wherein the TNFRSF agonist is added at a concentration sufficient to achieve a concentration in the cell culture medium of between 20 μg/mL and 40 μg/mL.

96. The process according to any one of claims 68 to 95, wherein IL-2 is present at an initial concentration of about 10 to about 6000 IU/mL in the first cell culture medium.

97. The process according to any one of claims 68 to 96, wherein IL-2 is present at an initial concentration of about 3000 IU/mL in the first cell culture medium.

98. The process according to any one of claims 68 to 97, wherein IL-2 is present at an initial concentration of about 800 to about 1100 R7/mL in the first cell culture medium.

99. The process according to any one of claims 68 to 98, wherein IL-2 is present at an initial concentration of about 1000 IU/mL in the first cell culture medium.

100. The process according to any one of claims 68 to 99, wherein IL-2 is present at an initial concentration of about 10 to about 6000 IU/mL in the second cell culture medium.

101. The process according to any one of claims 68 to 100, wherein IL-2 is present at an initial concentration of about 3000 IU/mL in the second cell culture medium.

102. The process according to any one of claims 68 to 101, wherein IL-2 is present at an initial concentration of about 800 to about 1100 IU/mL in the second cell culture medium.

103. The process according to any one of claims 68 to 102, wherein IL-2 is present at an initial concentration of about 1000 IU/mL in the second cell culture medium.

104. The process according to any one of claims 68 to 103, wherein IL-15 is present in the first cell culture medium.

105. The process according to any one of claims 68 to 104, wherein IL-15 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the first cell culture medium.

106. The process according to any one of claims 68 to 105, wherein IL-15 is present in the

second cell culture medium.

107. The process according to any one of claims 68 to 106, wherein IL-15 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the second cell culture medium.

108. The process according to any one of claims 68 to 107, wherein IL-21 is present in the first cell culture medium.

109. The process according to any one of claims 68 to 108, wherein IL-21 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the first cell culture medium.

110. The process according to any one of claims 68 to 109, wherein IL-21 is present in the

second cell culture medium.

111. The process according to any one of claims 68 to 110, wherein IL-21 is present at an initial concentration of about 5 ng/mL to about 20 ng/mL in the second cell culture medium.

112. The process according to any one of claims 68 to 111, wherein OKT-3 antibody is present at an initial concentration of about 10 ng/mL to about 60 ng/mL in the second cell culture medium.

113. The process according to any one of claims 68 to 112, wherein OKT-3 antibody is present at an initial concentration of about 30 ng/mL in the second cell culture medium.

114. The process according to any one of claims 68 to 113, wherein the initial expansion is performed using a gas permeable container.

115. The process according to any one of claims 68 to 114, wherein the rapid expansion is

performed using a gas permeable container.

116. A population of tumor infiltrating lymphocytes (TILs) obtainable from a process according to any one of claims 68 to 115.

117. A pharmaceutical composition comprising a population of tumor infiltrating lymphocytes (TILs) for use in treating a cancer wherein the population of tumor infiltrating lymphocytes (TILs) is obtainable by a process according to any one of claims 68 to 115, wherein

optionally the pharmaceutical composition comprises the third population of TILs.

118. The pharmaceutical composition for use in the treatment of a cancer according to claim 117, wherein the pharmaceutical composition is for use in combination with a T FRSF agonist.

119. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a TNFRSF agonist wherein the TNFRSF agonist is for administration on the day after administration of the third population of TILs to the patient, and wherein the TNFRSF agonist is administered intravenously at a dose of between 0.1 mg/kg and 50 mg/kg every four weeks for up to eight cycles.

120. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a TNFRSF agonist wherein the TNFRSF agonist is for administration prior to the step of resecting of a tumor from the patient, and wherein the TNFRSF agonist is for administration intravenously at a dose of between 0.1 mg/kg and 50 mg/kg every four weeks for up to eight cycles.

121. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 for use in combination with a non-myeloablative lymphodepletion regimen.

122. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a myeloablative lymphodepletion regimen prior to administering the third population of TILs to the patient.

123. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a myeloablative lymphodepletion regimen wherein the non-myeloablative lymphodepletion regimen is for administration prior to administering the third population of TILs to the patient, and wherein the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of 60 mg/m2/day for two days followed by administration of fludarabine at a dose of 25 mg/m2/day for five days.

124. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a decrescendo IL-2 regimen.

125. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a decrescendo IL-2 regimen starting on the day after administration of the third population of TILs to the patient, wherein the decrescendo IL-2 regimen comprises aldesleukin administered intravenously at a dose of 18,000,000 IU/m2 on day 1, 9,000,000 IU/m2 on day 2, and 4,500,000 IU/m2 on days 3 and 4.

126. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with pegylated IL-2.

127. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with pegylated IL-2 administered after administration of the third population of TILs to the patient at a dose of 0.10 mg/day to 50 mg/day.

128. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a high-dose IL-2 regimen.

129. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a high-dose IL-2 regimen starting on the day after administration of the third population of TILs to the patient.

130. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a high-dose IL-2 regimen starting on the day after administration of the third population of TILs to the patient, wherein the high-dose IL-2 regimen comprises 600,000 or 720,000 IU/kg of aldesleukin, or a biosimilar or variant thereof, administered as a 15-minute bolus intravenous infusion every eight hours until tolerance.

131. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor.

132. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

133. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor, wherein the PD-1 inhibitor or PD-L1 inhibitor is administered prior to resection of the tumor from the patient.

134. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor prior to resection of the tumor from the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

135. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor after resection a tumor from the patient.

136. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor after resection of the tumor from the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

137. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor which is for administration after administering the third population of TILs to the patient.

138. The pharmaceutical composition for use in the treatment of a cancer according to claim 117 wherein the pharmaceutical composition is for use in combination with a PD-1 inhibitor or PD-L1 inhibitor which is for administration after administering the third population of TILs to the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and fragments, derivatives, variants, biosimilars, and combinations thereof.

139. The pharmaceutical composition for use in the treatment of a cancer according to any one of claims 117 to 138 wherein the cancer is selected from the group consisting of melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer, renal cell carcinoma, acute myeloid leukemia, colorectal cancer,

cholangiocarcinoma, and sarcoma.

140. The pharmaceutical composition for use in the treatment of a cancer according to any one of claims 117 to 138, wherein the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC), triple negative breast cancer, double-refractory melanoma, and uveal (ocular) melanoma.

141. A method of treating a cancer with a population of tumor infiltrating lymphocytes (TILs) comprising the steps of:

(a) resecting a tumor from a patient;

(b) obtaining a first population of TILs from the tumor;

(c) performing an initial expansion of the first population of TILs in a first cell culture

medium to obtain a second population of TILs, wherein the second population of TILs is at least 5-fold greater in number than the first population of TILs, wherein the first cell culture medium comprises IL-2, and wherein the initial expansion is performed over a period of 11 days or less;

(d) performing a rapid expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs, wherein the third population of TILs is at least 50-fold greater in number than the second population of TILs after 7 days from the start of the rapid expansion; wherein the second cell culture medium comprises IL- 2, OKT-3 (anti-CD3) antibody, peripheral blood mononuclear cells (PBMCs), and a TNFRSF agonist, and wherein the rapid expansion is performed over a period of 11 days or less;

(e) harvesting the third population of TILs; and

(f) administering a therapeutically effective portion of the third population of TILs to the patient.

142. The method of Claim 141, wherein the TNFRSF agonist is selected from the group

consisting of a 4-1BB agonist, an OX40 agonist, and a combination thereof.

143. The method of Claim 142, wherein the TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is selected from the group consisting of urelumab, utomilumab, EU-101, a fusion protein, and fragments, derivatives, variants, biosimilars, and combinations thereof.

144. The method of Claim 142, wherein the TNFRSF agonist is a OX40 agonist, and the OX40 agonist is selected from the group consisting of tavolixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM- 18455, and fragments, derivatives, variants, biosimilars, and combinations thereof.

145. The method of any one of Claims 141 to 144, wherein the TNFRSF agonist is present at the start of step (d) at a concentration between 1 μg/mL and 30 μg/mL.

146. The method of Claim 145, wherein the TNFRSF agonist is present at the start of step (d) at a concentration between 5 μg/mL and 20 μg/mL.

147. The method of Claim 146, wherein the TNFRSF agonist is present at the start of step (d) at a concentration of about 10 μg/mL.

148. The method of any one of Claims 141 to 144, wherein the TNFRSF agonist is maintained throughout step (d) at a concentration between 1 μg/mL and 30 μg/mL.

149. The method of Claim 145, wherein the TNFRSF agonist is maintained throughout step (d) at a concentration between 5 μg/mL and 20 μg/mL.

150. The method of Claim 146, wherein the TNFRSF agonist is maintained throughout step (d) at a concentration of about 10 μg/mL.

151. The method of any one of Claims 141 to 150, wherein the third population of TILs exhibits an increased ratio of CD8+ TILs to CD4+ TILs in comparison to the reference ratio of CD8+ TILs to CD4+ TILs in the second population of TILs.

152. The method of Claim 151, wherein the increased ratio is at least 5% greater than the

reference ratio.

153. The method of Claim 152, wherein the increased ratio is at least 10% greater than the

reference ratio.

154. The method of Claim 153, wherein the increased ratio is at least 20% greater than the

reference ratio.

155. The method of Claim 154, wherein the increased ratio is at least 35% greater than the

reference ratio.

156. The method of Claim 155, wherein the increased ratio is at least 50% greater than the

reference ratio.

157. The method of any one of Claims 141 to 156, wherein the cancer is selected from the group consisting of melanoma, uveal (ocular) melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer (head and neck squamous cell cancer), renal cell carcinoma, colorectal cancer, pancreatic cancer, glioblastoma, cholangiocarcinoma, and sarcoma.

158. The method of any one of Claims 141 to 156, wherein the cancer is selected from the group consisting of cutaneous melanoma, uveal (ocular) melanoma, platinum -resistant ovarian cancer, pancreatic ductal adenocarcinoma, osteosarcoma, triple-negative breast cancer, and non-small-cell lung cancer.