Processes for production of tumor infiltrating lymphocytes and uses of same in immunotherapy

Patent metadata

Field	Value
Publication number	US-11517592-B1
Application number	US-202217856800-A
Country	US
Kind code	B1
Filing date	Jul 1, 2022
Priority date	Mar 29, 2017
Publication date	Dec 6, 2022
Grant date	Dec 6, 2022

Abstract

The present invention provides improved and/or shortened methods for expanding TILs and producing therapeutic populations of TILs, including novel methods for expanding TIL populations in a closed system that lead to improved efficacy, improved phenotype, and increased metabolic health of the TILs in a shorter time period, while allowing for reduced microbial contamination as well as decreased costs. Such TILs find use in therapeutic treatment regimens.

First claim

What is claimed is:

1. A method for treating a subject with melanoma, the method comprising administering expanded tumor infiltrating lymphocytes (TILs) comprising:
(a) adding a tumor digest or tumor fragments into a closed system, wherein the tumor digest or tumor fragments comprise a first population of TILs and are obtained from a tumor that was resected from the subject;
(b) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for a first period of about 3 to 11 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) occurs without opening the system;
(c) performing a second expansion by supplementing the cell culture medium with additional IL-2, OKT-3, and antigen presenting cells (APCs) to produce a third population of TILs, wherein the second expansion is performed for a second period of about 7 to 11 days to obtain the third population of TILs, wherein the second expansion is performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (b) to step (c) occurs without opening the system;
(d) harvesting the third population of TILs obtained from step (c), wherein the transition from step (c) to step (d) occurs without opening the system;
(e) transferring the harvested third TIL population from step (d) to an infusion bag, wherein the transfer from step (d) to (e) occurs without opening the system;
(f) cryopreserving the infusion bag comprising the harvested TIL population from step (e) using a cryopreservation process, wherein no selection of the first population of TILs, second population of TILs, third population of TILs, harvested TIL population, and/or the therapeutic TIL population based on CD8 expression is performed during any of steps (a) to step (0, and
(g) administering a therapeutically effective dosage of the harvested TIL population from the infusion bag in step (f) to the subject.

2. The method according to claim 1 , wherein the tumor digest in step (a) was prepared by incubating a sample of the tumor that was resected from the subject in an enzymatic media.

3. The method according to claim 2 , wherein the preparation of the tumor digest comprises disrupting the tumor sample mechanically so as to dissociate the tumor sample, optionally further comprising purifying the disassociated tumor sample using a density gradient separation.

4. The method according to claim 2 , wherein the enzymatic media comprises DNase.

5. The method according to claim 4 , wherein the enzymatic media comprises 30 units/mL of DNase.

6. The method according to claim 2 , wherein the enzymatic media comprises collagenase.

7. The method according to claim 6 , wherein the enzymatic media comprises 1.0 mg/mL of collagenase.

8. The method according to claim 1 , wherein the third population of TILs harvested in step (d) comprises sufficient TILs for administering a therapeutically effective dosage of the TILs in step (g).

9. The method according to claim 8 , wherein the therapeutically effective dosage in step (g) comprises from about 1×10 9 to 5×10 10 TILs.

10. The method according to claim 8 , wherein the therapeutically effective dosage in step (g) comprises from about 1×10 9 to about 9×10 10 TILs.

11. The method according to claim 8 , wherein the therapeutically effective dosage in step (g) comprises from about 5×10 9 to 1×10 10 TILs.

12. The method according to claim 8 , wherein the therapeutically effective dosage in step (g) comprises from about 1×10 10 to 5×10 10 TILs.

13. The method according to claim 1 , wherein the APCs comprise peripheral blood mononuclear cells (PBMCs).

14. The method according to claim 13 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.

15. The method according to claim 1 , wherein prior to administering a therapeutically effective dosage of TIL cells in step (g), a non-myeloablative lymphodepletion regimen has been administered to the subject.

16. The method according to claim 15 , where the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of 60 mg/m 2 /day for two days followed by administration of fludarabine at a dose of 25 mg/m 2 /day for five days.

17. The method according to claim 1 , further comprising the step of treating the subject with a high-dose IL-2 regimen starting on the day after administration of the TIL cells to the subject in step (g).

18. The method according to claim 17 , wherein the high-dose IL-2 regimen comprises 600,000 or 720,000 IU/kg administered as a 15-minute bolus intravenous infusion every eight hours until tolerance.

19. The method according to claim 1 , wherein the melanoma is metastatic melanoma.

20. The method according to claim 1 , wherein the first expansion in step (b) and the second expansion in step (c) are each individually performed within a period of 11 days.

21. The method according to claim 1 , wherein the first expansion is performed for about 10 days or for about 11 days.

22. The method according to claim 1 , wherein the second expansion is performed for about 10 days or for about 11 days.

23. The method according to claim 1 , wherein steps (a) through (e) are performed in about 10 days to about 22 days.

24. The method according to claim 1 , wherein steps (a) through (e) are performed in about 20 days or less.

25. The method according to claim 1 , wherein no selection of the first population of TILs, second population of TILs, third population of TILs, harvested TIL population, and/or the therapeutic TIL population based on CD4 and/or CD8 expression is performed during any of steps (a) to step (f).

26. The method according to claim 1 , wherein no selection of the first population of TILs, second population of TILs, third population of TILs, harvested TIL population, and/or the therapeutic TIL population is performed during any of steps (a) to step (f).

27. The method according to claim 1 , wherein the therapeutic population of TILs is capable of exhibiting IFN-gamma (IFN-γ) secretion greater than 1000 pg/ml.

Assignees

• Iovance Biotherapeutics Inc

Inventors

• Wardell Seth
• Bender James
• Lotze Michael T

Classifications

• C12N2501/2321

  Interleukin-21 (IL-21) · CPC title

• C12N2501/24

  Interferons [IFN] · CPC title

• C12N2502/11

  blood or immune system cells · CPC title

• A61K38/217

  IFN-gamma · CPC title

• C12N2501/603

  Oct-3/4 · CPC title