Granted Patent B2

US 12,611,428 · App. 17/110,179 · Granted Apr 28, 2026

Inventors: Seth Wardell (Tampa, FL); James Bender (Rancho Santa Margarita, CA)

Assignee: Iovance Biotherapeutics, Inc.

A61K35/17A01N1/162A61K9/0019A61K31/675A61K31/7076A61K38/2013A61K40/11A61K40/428A61P35/00C12N5/0634C12N5/0636C12N5/0638A61K38/217A61K39/0011A61K2039/5154A61K2039/5156A61K2039/5158A61K2039/55533A61K40/50C12N2501/04C12N2501/2302C12N2501/2315C12N2501/2321C12N2501/24C12N2501/603C12N2502/11C12N2506/30

Abstract

Claims (63)

1 . A method for treating a subject with a cancer, the method comprising administering a combination of expanded tumor infiltrating lymphocytes (TILs) and an anti-PD-1 antibody, the method 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, 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 about 3-14 days to obtain the second population of TILs, 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, optionally OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 4-6 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 (c) to step (d) occurs without opening the system;

(e) dividing the third population of TILs into a first plurality of 2-5 subpopulations of TILs, wherein at least 1.0×10 9 TILs are present in each subpopulation, wherein the transition from step (d) to (e) occurs without opening the system;

(f) performing a third expansion of the first plurality of subpopulations of TILs by supplementing the cell culture medium of each subpopulation of TILs with additional IL-2, optionally OKT-3, to produce a second plurality of subpopulations of TILs, wherein the third expansion is performed for about 5-7 days, wherein the third expansion for each subpopulation is performed in a closed container providing a third gas-permeable surface area, and wherein the transition from step (e) to step (f) occurs without opening the system;

(g) harvesting the second plurality of subpopulations of TILs obtained from step (f), wherein the transition from step (f) to step (g) occurs without opening the system; and

(h) transferring the harvested subpopulations of TILs from step (g) to one or more infusion bags, wherein the transition from step (g) to (h) occurs without opening the system;

(i) cryopreserving the one or more infusion bags comprising the harvested subpopulations of TILs from step (h) using a cryopreservation process; and

(j) administering to the subject:

i) a therapeutically effective dosage of the harvested subpopulations of TILs from the infusion bag in step (g); and

ii) the anti-PD-1 antibody.

2 . The method according to claim 1 , wherein the cell culture medium is CTS Optimizer.

3 . The method according to claim 1 , wherein the second plurality of subpopulations of TILs harvested in step (g) comprises sufficient TILs for administering a therapeutically effective dosage of the TILs in step (j).

4 . The method according to claim 3 , wherein the therapeutically effective dosage in step (j) comprises from about 1×10 9 to about 9×10 10 TILs.

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

6 . The method according to claim 1 , wherein the third population of TILs harvested in step (e) exhibits an increased subpopulation of CD8+ cells relative to the first and/or second population of TILs.

7 . The method according to claim 5 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.

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

9 . The method according to claim 8 , where the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of mg/kg/day for two days followed by administration of fludarabine at a dose of 25 mg/m 2 /day for five days.

10 . 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 (j).

11 . The method according to claim 10 , 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.

12 . A method according to claim 1 , wherein the second plurality of subpopulations of TILs in step (f) provides for increased efficacy, increased interferon-gamma (IFN-γ) production, increased polyclonality, increased average IP-10, and/or increased average MCP-1 when administered to the subject.

13 . The method according to claim 1 , wherein the cancer is selected from the group consisting of melanoma (including metastatic melanoma), ovarian cancer, cervical cancer, non-small-cell lung cancer (NSCLC), lung cancer, bladder cancer, breast cancer, cancer caused by human papilloma virus, head and neck cancer (including head and neck squamous cell carcinoma (HNSCC)), renal cancer, and renal cell carcinoma.

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

15 . The method according to claim 1 , wherein the anti-PD-1 antibody is nivolumab.

16 . The method according to claim 1 , wherein the anti-PD-1 antibody is pembrolizumab.

17 . A method for treating a subject with a cancer, the method comprising administering a combination of expanded tumor infiltrating lymphocytes (TILs) and an anti-PD-1 antibody, the method 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 a tumor digest;

(b) adding the tumor digest into a closed system;

(c) 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 about 3-14 days to obtain the second population of TILs, 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, optionally OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 4-6 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 (c) to step (d) occurs without opening the system;

(e) dividing the third population of TILs into a first plurality of 2-5 subpopulations of TILs, wherein at least 1.0×10 9 TILs are present in each subpopulation, wherein the transition from step (d) to (e) occurs without opening the system;

(f) performing a third expansion of the first plurality of subpopulations of TILs by supplementing the cell culture medium of each subpopulation of TILs with additional IL-2, optionally OKT-3, to produce a second plurality of subpopulations of TILs, wherein the third expansion is performed for about 5-7 days, wherein the third expansion for each subpopulation is performed in a closed container providing a third gas-permeable surface area, and wherein the transition from step (e) to step (f) occurs without opening the system;

(g) harvesting the second plurality of subpopulations of TILs obtained from step (f), wherein the transition from step (f) to step (g) occurs without opening the system; and

(h) transferring the harvested subpopulations of TILs from step (g) to one or more infusion bags, wherein the transition from step (g) to (h) occurs without opening the system;

(i) cryopreserving the one or more infusion bags comprising the harvested subpopulations of TILs from step (h) using a cryopreservation process; and

(j) administering to the subject:

i) a therapeutically effective dosage of the harvested subpopulations of TILs from the infusion bag in step (g); and

ii) the anti-PD-1 antibody.

18 . The method according to claim 17 , wherein processing a tumor sample obtained from the subject into a tumor digest in step (a) comprises incubating the tumor sample in an enzymatic media.

19 . The method according to claim 18 , wherein processing a tumor sample obtained from the subject into a tumor digest in step (a) further comprises disrupting the tumor sample mechanically so as to dissociate the tumor sample.

20 . The method according to claim 19 , wherein processing a tumor sample obtained from the subject into a tumor digest in step (a) further comprises purifying the disassociated tumor sample using a density gradient separation.

21 . The method according to claim 18 , wherein the enzymatic media comprises DNase.

22 . The method according to claim 21 , wherein the enzymatic media comprises 30 units/mL of DNase.

23 . The method according to claim 18 , wherein the enzymatic media comprises collagenase.

24 . The method according to claim 23 , wherein the enzymatic media comprises 1.0 mg/mL of collagenase.

25 . The method according to claim 17 , wherein the cell culture medium is CTS Optimizer.

26 . The method according to claim 17 , wherein the second plurality of subpopulations of TILs harvested in step (g) comprises sufficient TILs for administering a therapeutically effective dosage of the TILs in step (j).

27 . The method according to claim 19 , wherein the therapeutically effective dosage in step (j) comprises from about 1×10 9 to about 9×10 10 TILs.

28 . The method according to claim 17 , wherein the APCs comprise peripheral blood mononuclear cells (PBMCs).

29 . The method according to claim 17 , wherein the third population of TILs harvested in step (e) exhibits an increased subpopulation of CD8+ cells relative to the first and/or second population of TILs.

30 . The method according to claim 21 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.

31 . The method according to claim 17 , wherein prior to administering a therapeutically effective dosage of TIL cells in step (j), a non-myeloablative lymphodepletion regimen has been administered to the subject.

32 . The method according to claim 24 , 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.

33 . The method according to claim 17 , 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 (j).

34 . The method according to claim 26 , 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.

35 . The method according to claim 17 , wherein the second plurality of subpopulations of TILs in step (f) provides for increased efficacy, increased interferon-gamma (IFN-γ) production, increased polyclonality, increased average IP-10, and/or increased average MCP-1 when administered to the subject.

36 . The method according to claim 17 , wherein the cancer is selected from the group consisting of melanoma (including metastatic melanoma), ovarian cancer, cervical cancer, non-small-cell lung cancer (NSCLC), lung cancer, bladder cancer, breast cancer, cancer caused by human papilloma virus, head and neck cancer (including head and neck squamous cell carcinoma (HNSCC)), renal cancer, and renal cell carcinoma.

37 . The method according to claim 17 , wherein steps (a) through (h) are performed in about 10 days to about 22 days.

38 . The method according to claim 17 , wherein the anti-PD-1 antibody is nivolumab.

39 . The method according to claim 17 , wherein the anti-PD-1 antibody is pembrolizumab.

Assignments (1)

Continuity (12)

References Cited (395)