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

Patent metadata

Field	Value
Publication number	US-12473532-B2
Application number	US-201917053344-A
Country	US
Kind code	B2
Filing date	May 9, 2019
Priority date	May 10, 2018
Publication date	Nov 18, 2025
Grant date	Nov 18, 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 method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:
(a) pre-treating a patient with a regimen comprising a kinase inhibitor or an ITK inhibitor; (b) obtaining a first population of TILs from a tumor resected from a patient by processing a tumor sample obtained from the patient into multiple tumor fragments; (c) adding the tumor fragments into a closed system; (d) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2, and optionally OKT-3, 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 3-14 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) occurs without opening the system; (e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, optionally 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 7-14 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 (d) to step (e) occurs without opening the system; (f) harvesting the therapeutic population of TILs obtained from step (e), wherein the transition from step (e) to step (f) occurs without opening the system; and (g) transferring the harvested TIL population from step (f) to an infusion bag, wherein the transfer from step (f) to (g) occurs without opening the system.

2 . The method according to claim 1 , further comprising the step of cryopreserving the infusion bag comprising the harvested TIL population in step (g) using a cryopreservation process.

3 . The method according to claim 1 , wherein the cryopreservation process is performed using a 1:1 ratio of harvested TIL population to cryopreservation media.

4 . The method according to claim 1 , wherein the antigen-presenting cells are peripheral blood mononuclear cells (PBMCs).

5 . The method according to claim 4 , wherein the PBMCs are irradiated and allogeneic.

6 . The method according to claim 4 , wherein the PBMCs are added to the cell culture on any of days 9 through 14 in step (e).

7 . The method according to claim 1 , wherein the antigen-presenting cells are artificial antigen-presenting cells.

8 . The method according to claim 1 , wherein the multiple fragments comprise about 4 to about 50 fragments, wherein each fragment has a volume of about 27 mm 3 .

9 . The method according to claim 1 , wherein the multiple fragments comprise about 30 to about 60 fragments with a total volume of about 1300 mm 3 to about 1500 mm 3 .

10 . The method according to claim 9 , wherein the multiple fragments comprise about 50 fragments with a total volume of about 1350 mm 3 .

11 . The method according to claim 1 , wherein the multiple fragments comprise about 50 fragments with a total mass of about 1 gram to about 1.5 grams.

12 . The method according to claim 1 , wherein the cell culture medium in step (d) further comprises IL-15 and/or IL-21.

13 . The method according to claim 1 , wherein the IL-2 concentration is about 10,000 IU/mL to about 5,000 IU/mL.

14 . The method according to claim 12 , wherein the IL-15 concentration is about 500 IU/mL to about 100 IU/mL.

15 . The method according to claim 12 , wherein the IL-21 concentration is about 20 IU/mL to about 0.5 IU/mL.

16 . The method according to claim 3 , wherein the cryopreservation media comprises dimethlysulfoxide (DMSO).

17 . The method according to claim 3 , wherein the cryopreservation media comprises 7% to 10% DMSO.

18 . The method according to claim 1 , wherein the first period in step (d) and the second period in step (f) are each individually performed within a period of 10 days, 11 days, or 12 days.

19 . The method according to claim 1 , wherein steps (b) through (g) are performed within a period of about 10 days to about 22 days.

20 . The method according to claim 1 , wherein steps (b) through (g) are performed in 10 days or less.

21 . The method according to claim 1 , wherein the therapeutic population of TILs harvested in step (f) comprises sufficient TILs for a therapeutically effective dosage of the TILs.

22 . The method according to claim 21 , wherein the number of TILs sufficient for a therapeutically effective dosage is from about 2.3×10 10 to about 13.7×10 10 .

23 . The method according to claim 1 , wherein steps (c) through (f) are performed in a single container, wherein performing steps (c) through (f) in a single container results in an increase in TIL yield per resected tumor as compared to performing steps (c) through (f) in more than one container.

24 . The method according to claim 1 , wherein the TILs from step (g) are infused into a patient.

25 . The method according to claim 1 , wherein the multiple fragments comprise about 4 fragments.

26 . A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:
(a) adding into a closed system processed tumor fragments from a tumor resected from a patient to obtain a first population of TILs, wherein the patient was pre-treated with a regimen comprising a kinase inhibitor or an ITK inhibitor prior to resection; (b) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2, and optionally OKT-3, 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 3-14 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 of the second population of TILs with additional IL-2, optionally 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 7-14 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; and (e) transferring the harvested TIL population from step (d) to an infusion bag, wherein the transfer from step (d) to (e) occurs without opening the system.

27 . A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:
(a) pre-treating a patient with a regimen comprising a kinase inhibitor or an ITK inhibitor; (b) obtaining a first population of TILs from a tumor resected from a patient by processing a tumor sample o

Assignees

• Iovance Biotherapeutics Inc

Inventors

• Wardell Seth

Classifications

• C12N2501/998

  Proteins not provided for elsewhere · CPC title

• C12N2501/2321

  Interleukin-21 (IL-21) · CPC title

• C12N2501/2315

  Interleukin-15 (IL-15) · CPC title

• C12N2501/2302

  Interleukin-2 (IL-2) · CPC title

• C12N5/0638

  Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK] · CPC title