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

Patent metadata

Field	Value
Publication number	US-12194061-B2
Application number	US-202217817207-A
Country	US
Kind code	B2
Filing date	Aug 3, 2022
Priority date	Mar 29, 2017
Publication date	Jan 14, 2025
Grant date	Jan 14, 2025

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 cryopreserved therapeutic population of tumor infiltrating lymphocytes (TILs) produced by a method 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 a 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 third population of TILs is a therapeutic 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 therapeutic 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 therapeutic population of TILs from step (d) to an infusion bag, wherein the transfer from step (d) to (e) occurs without opening the system; and
(f) cryopreserving the infusion bag comprising the harvested therapeutic population of TILs from step (e) using a cryopreservation process.

2. The cryopreserved therapeutic population of TILs 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 cryopreserved therapeutic population of TILs according to claim 2 , further comprising disrupting the tumor sample mechanically so as to dissociate the tumor sample.

4. The cryopreserved therapeutic population of TILs according to claim 3 , further comprising purifying the disassociated tumor sample using a density gradient separation.

5. The cryopreserved therapeutic population of TILs according to claim 2 , wherein the enzymatic media comprises DNase.

6. The cryopreserved therapeutic population of TILs according to claim 5 , wherein the enzymatic media comprises 30 units/mL of DNase.

7. The cryopreserved therapeutic population of TILs according to claim 2 , wherein the enzymatic media comprises collagenase.

8. The cryopreserved therapeutic population of TILs according to claim 7 , wherein the enzymatic media comprises 1.0 mg/mL of collagenase.

9. The cryopreserved therapeutic population of TILs according to claim 1 , wherein the medium in the first expansion and/or the second expansion is free of human serum.

10. The cryopreserved therapeutic population of TILs according to claim 1 , wherein the therapeutic population of TILs harvested in step (d) comprises sufficient TILs for use in administering a therapeutically effective dosage to a subject.

11. The cryopreserved therapeutic population of TILs according to claim 10 , wherein the number of TILs sufficient for administering a therapeutically effective dosage is from about 1×10 9 to about 5×10 9 .

12. The cryopreserved therapeutic population of TILs according to claim 10 , wherein the number of TILs sufficient for administering a therapeutically effective dosage is from about 1×10 9 to about 9×10 10 .

13. The cryopreserved therapeutic population of TILs according to claim 10 , wherein the number of TILs sufficient for administering a therapeutically effective dosage is from about 5×10 9 to about 1×10 10 .

14. The cryopreserved therapeutic population of TILs according to claim 10 , wherein the number of TILs sufficient for administering a therapeutically effective dosage is from about 1×10 10 to about 5×10 10 .

15. The cryopreserved therapeutic population of TILs according to claim 1 , wherein the APCs are peripheral blood mononuclear cells (PBMCs).

16. The cryopreserved therapeutic population of TILs according to claim 15 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.

17. The cryopreserved therapeutic population of TILs according to claim 1 , wherein the therapeutic population of TILs harvested in step (d) exhibits an increased subpopulation of CD8+ cells relative to the first and/or second population of TILs.

18. The cryopreserved therapeutic population of TILs 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.

19. The cryopreserved therapeutic population of TILs according to claim 1 , wherein steps (a) through (e) are performed in about 10 days to about 22 days.

20. The cryopreserved therapeutic population of TILs according to claim 1 , wherein steps (a) through (e) are performed in about 15 days to about 22 days.

21. The cryopreserved therapeutic population of TILs according to claim 1 , wherein steps (a) through (e) are performed in about 20 days to about 22 days.

22. The cryopreserved therapeutic population of TILs according to claim 1 , wherein the tumor digest is a cryopreserved tumor digest.

Assignees

• Iovance Biotherapeutics Inc

Inventors

• Wardell Seth
• Bender James
• Lotze Michael T

Classifications

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A01N1/162

  Temperature processes, e.g. following predefined temperature changes over time · CPC title

• C12N2501/2321

  Interleukin-21 (IL-21) · CPC title

• C12N2501/2315

  Interleukin-15 (IL-15) · CPC title

• C12N5/0636

  T lymphocytes · CPC title