Search

US-12622927-B2 - Processes for production of tumor infiltrating lymphocytes and uses of same in immunotherapy

US12622927B2US 12622927 B2US12622927 B2US 12622927B2US-12622927-B2

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.

Inventors

  • Seth Wardell
  • James Bender
  • Michael T. Lotze

Assignees

  • IOVANCE BIOTHERAPEUTICS, INC.

Dates

Publication Date
20260512
Application Date
20220803

Claims (20)

  1. 1 . A method for treating a subject with a melanoma, the method comprising administering expanded tumor infiltrating lymphocytes (TILs) comprising: (a) performing a first expansion by culturing a first population of TILs from tumor fragments obtained from a tumor resected from the subject 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 11 days to obtain the second population of TILs; (b) 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 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 (a) to step (b) occurs without opening the system; (c) harvesting the third population of TILs obtained from step (b), wherein the transition from step (b) to step (c) occurs without opening the system; (d) transferring the harvested third TIL population from step (c) to an infusion bag, wherein the transfer from step (c) to (d) occurs without opening the system; (e) cryopreserving the infusion bag comprising the harvested TIL population from step (d) using a cryopreservation process; and (f) administering a therapeutically effective dosage of the harvested TIL population from the infusion bag in step (e) to the subject.
  2. 2 . The method according to claim 1 , wherein the third population of TILs harvested in step (c) comprises sufficient TILs for administering a therapeutically effective dosage of the TILs in step (f).
  3. 3 . The method according to claim 2 , wherein the therapeutically effective dosage in step (f) comprises from about 1×10 9 to about 9×10 10 TILs.
  4. 4 . The method according to claim 2 , wherein the therapeutically effective dosage in step (f) comprises from about 1×10 9 to about 5×10 9 TILs.
  5. 5 . The method according to claim 2 , wherein the therapeutically effective dosage in step (f) comprises from about 5×10 9 to about 1×10 10 TILs.
  6. 6 . The method according to claim 2 , wherein the therapeutically effective dosage in step (f) comprises from about 1×10 10 to about 5×10 10 TILs.
  7. 7 . The method according to claim 1 , wherein the APCs comprise peripheral blood mononuclear cells (PBMCs).
  8. 8 . The method according to claim 7 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.
  9. 9 . The method according to claim 1 , wherein prior to administering a therapeutically effective dosage of TIL cells in step (f), a non-myeloablative lymphodepletion regimen has been administered to the subject.
  10. 10 . The method according to claim 9 , where the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of 60 mg/kg/day for two days followed by administration of fludarabine at a dose of 25 mg/m 2 /day for five days.
  11. 11 . 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 (f).
  12. 12 . The method according to claim 11 , 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.
  13. 13 . The method according to claim 1 , wherein the melanoma is metastatic melanoma.
  14. 14 . The method according to claim 1 , wherein the closed container in step (a) and/or step (b) is a gas-permeable bag.
  15. 15 . The method according to claim 1 , wherein the tumor fragments comprise about 4 to about 50 fragments, wherein each fragment has a volume of about 27 mm 3 .
  16. 16 . The method according to claim 1 , wherein the tumor fragments comprise about 30 to about 60 fragments with a total volume of about 1300 mm 3 to about 1500 mm 3 .
  17. 17 . The method according to claim 1 , wherein the tumor fragments comprise about 50 fragments with a total mass of about 1 gram to about 1.5 grams.
  18. 18 . The method according to claim 1 , wherein the cell culture medium in step (b) further comprises IL-15 and/or IL-21.
  19. 19 . The method according to claim 17 , wherein the IL-15 concentration is about 500 IU/mL to about 100 IU/mL.
  20. 20 . The method according to claim 17 , wherein the IL-21 concentration is about 20IU/mL to about 0.5 IU/mL.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. patent application Ser. No. 17/856,806, filed on Jul. 1, 2022, which is a continuation of U.S. patent application Ser. No. 17/147,080, filed on Jan. 12, 2021, which is a divisional of U.S. patent application Ser. No. 15/863,634, filed on Jan. 5, 2018, now U.S. Pat. No. 10,894,063, which claims priority to U.S. Provisional Patent Application No. 62/478,506, filed on Mar. 29, 2017, U.S. Provisional Patent Application No. 62/539,410, filed on Jul. 31, 2017, U.S. Provisional Patent Application No. 62/548,306, filed on Aug. 21, 2017, U.S. Provisional Patent Application No. 62/554,538, filed on Sep. 5, 2017, U.S. Provisional Patent Application No. 62/559,374, filed on Sep. 15, 2017, U.S. Provisional Patent Application No. 62/567,121, filed on Oct. 2, 2017, U.S. Provisional Patent Application No. 62/577,655, filed on Oct. 26, 2017, U.S. Provisional Patent Application No. 62/582,874, filed on Nov. 7, 2017, and U.S. Provisional Patent Application No. 62/596,374, filed on Dec. 8, 2017, which are hereby incorporated by reference in their entirety. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML file, created on Aug. 2, 2022, is named 116983-5017-US_Sequence_Listing.xml and is 16,384 bytes in size. BACKGROUND OF THE INVENTION Treatment of bulky, refractory cancers using adoptive transfer of tumor infiltrating lymphocytes (TILs) represents a powerful approach to therapy for patients with poor prognoses. Gattinoni, et al., Nat. Rev. Immunol. 2006, 6, 383-393. A large number of TILs are required for successful immunotherapy, and a robust and reliable process is needed for commercialization. This has been a challenge to achieve because of technical, logistical, and regulatory issues with cell expansion. IL-2-based TIL expansion followed by a “rapid expansion process” (REP) has become a preferred method for TIL expansion because of its speed and efficiency. Dudley, et al., Science 2002, 298, 850-54; Dudley, et al., J. Clin. Oncol. 2005, 23, 2346-57; Dudley, et al., J. Clin. Oncol. 2008, 26, 5233-39; Riddell, et al., Science 1992, 257, 238-41; Dudley, et al., J. Immunother. 2003, 26, 332-42. REP can result in a 1,000-fold expansion of TILs over a 14-day period, although it requires a large excess (e.g., 200-fold) of irradiated allogeneic peripheral blood mononuclear cells (PBMCs, also known as mononuclear cells (MNCs)), often from multiple donors, as feeder cells, as well as anti-CD3 antibody (OKT3) and high doses of IL-2. Dudley, et al., J. Immunother. 2003, 26, 332-42. TILs that have undergone an REP procedure have produced successful adoptive cell therapy following host immunosuppression in patients with melanoma. Current infusion acceptance parameters rely on readouts of the composition of TILs (e.g., CD28, CD8, or CD4 positivity) and on fold expansion and viability of the REP product. Current TIL manufacturing processes are limited by length, cost, sterility concerns, and other factors described herein such that the potential to commercialize such processes is severely limited, and for these and other reasons, at the present time no commercial process has become available. There is an urgent need to provide TIL manufacturing processes and therapies based on such processes that are appropriate for commercial scale manufacturing and regulatory approval for use in human patients at multiple clinical centers. BRIEF SUMMARY OF THE INVENTION The present invention provides improved and/or shortened methods for expanding TILs and producing therapeutic populations of TILs. The present invention provides a method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising: (a) 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;(b) adding the tumor fragments into a closed system;(c) 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 about 3-14 days to obtain the second population of TILs, wherein the second population of TILs is at least 50-fold greater in number than the first population of TILs, and wherein the transition from step (b) to step (c) occurs without opening the system;(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs 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 about 7-14 days to obtain the third population of TILs, wherein the third