Methods of using tumor infiltrating lymphocytes in double-refractory melanoma

Abstract

Methods of treating melanomas refractory to other therapies using tumor infiltrating lymphocytes are disclosed.

We claim: 
     
         1 . A method of treating double-refractory metastatic melanoma in a patient in need thereof, the method comprising administering a therapeutically effective population of tumor infiltrating lymphocytes (TILs) to the patient. 
     
     
         2 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is a cutaneous double-refractory metastatic melanoma. 
     
     
         3 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to at least two prior systemic treatment courses, not including neo-adjuvant or adjuvant therapies. 
     
     
         4 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to aldesleukin or a biosimilar thereof. 
     
     
         5 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to pembrolizumab or a biosimilar thereof. 
     
     
         6 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to nivolumab or a biosimilar thereof. 
     
     
         7 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to ipilimumab or a biosimilar thereof. 
     
     
         8 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to ipilimumab or a biosimilar thereof and pembrolizumab or a biosimilar thereof. 
     
     
         9 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to ipilimumab or a biosimilar thereof and nivolumab or a biosimilar thereof. 
     
     
         10 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to a BRAF inhibitor. 
     
     
         11 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to a PD-L1 inhibitor. 
     
     
         12 . The method of  claim 11 , wherein the PD-L1 inhibitor is selected from the group consisting of avelumab, atezolizumab, durvalumab, and biosimilars thereof. 
     
     
         13 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to a combination of a PD-1 inhibitor and a CTLA-4 inhibitor. 
     
     
         14 . The method of  claim 13 , wherein the PD-1 inhibitor is nivolumab or a biosimilar thereof and the CTLA-4 inhibitor is selected from the group consisting of ipilumumab, tremelimumab, and biosimilars thereof. 
     
     
         15 . The method of  claim 1 , wherein the double-refractory metastatic melanoma is refractory to a combination of a BRAF inhibitor and a MEK inhibitor. 
     
     
         16 . The method of  claim 15 , wherein the BRAF inhibitor is dabrafenib or a pharmaceutically-acceptable salt thereof and the MEK inhibitor is trametinib or a pharmaceutically-acceptable salt or solvate thereof. 
     
     
         17 . The method of  claim 1 , wherein the metastatic melanoma is resistant to a PD-1 inhibitor or PD-L1 inhibitor. 
     
     
         18 . The method of  claim 17 , wherein the PD-1 or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, avelumab, atezolizumab, durvalumab, and biosimilars thereof. 
     
     
         19 . The method of  claims 1 - 10 , wherein the patient does not possess a BRAF mutation. 
     
     
         20 . The method of  claim 1 , wherein the patient has received at most 4 doses of nivolumab or a biosimilar thereof prior to receiving the therapeutically effective population of TILs. 
     
     
         21 . The method of  claims 1 - 12 , wherein the patient has progressed or had no response to at least two prior systemic treatment courses. 
     
     
         22 . A method of treating double-refractory metastatic melanoma in a patient in need thereof, the method comprising:
 (a) obtaining a first population of TILs from a tumor resected from the 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 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, optionally 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 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 (c) to step (d) occurs without opening the system;   (e) harvesting the therapeutic population of TILs obtained from step (d) to provide a harvested TIL population, wherein the transition from step (d) to step (e) occurs without opening the system;   (f) transferring the harvested TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) occurs without opening the system, and optionally cryopreserving the harvested TIL population and   (g) administering a therapeutically effective amount of the harvested TIL population to the patient with double-refractory metastatic melanoma.   
     
     
         23 . The method of  claim 20 , wherein the patient has been previously treated with a PD-1 inhibitor or a biosimilar thereof. 
     
     
         24 . The method of  claim 21 , wherein the PD-1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, and biosimilars thereof. 
     
     
         25 . The method of  claim 20 , wherein the patient has been previously treated with a PD-L1 inhibitor or a biosimilar thereof. 
     
     
         26 . The method of  claim 25 , wherein the PD-L1 inhibitor is selected from the group consisting of avelumab, atezolizumab, durvalumab, and biosimilars thereof. 
     
     
         27 . The method of any one of  claims 23 - 24 , wherein the PD-1 inhibitor or a biosimilar thereof was co-administered with a CTLA-4 inhibitor or biosimilar thereof. 
     
     
         28 . The method of any one of  claims 25 - 26 , wherein the PD-L1 inhibitor or a biosimilar thereof was co-administered with a CTLA-4 inhibitor or biosimilar thereof 
     
     
         29 . The method of any one of  claims 22 - 28 , wherein the patient has been previously treated with one additional prior line of systemic therapy. 
     
     
         30 . The method of  claim 29 , wherein the one additional prior line of systemic therapy is a BRAF inhibitor or a pharmaceutically-acceptable salt thereof 
     
     
         31 . The method of  claim 30 , wherein the BRAF inhibitor is selected from the group consisting of vemurafenib, dabrafenib, and pharmaceutically-acceptable salts thereof 
     
     
         32 . The method of  claim 29 , wherein the one additional prior line of systemic therapy is a MEK inhibitor or a pharmaceutically-acceptable salt or solvate thereof 
     
     
         33 . The method of  claim 32 , wherein the MEK inhibitor is selected from the group consisting of trametinib, cobimetinib, and pharmaceutically-acceptable salts or solvates thereof 
     
     
         34 . The method of  claim 29 , wherein the one additional prior line of systemic therapy is a combination of a BRAF inhibitor or a pharmaceutically-acceptable salt thereof and a MEK inhibitor or a pharmaceutically-acceptable salt or solvate thereof 
     
     
         35 . The method of  claim 34 , wherein the BRAF inhibitor is selected from the group consisting of vemurafenib, dabrafenib, and pharmaceutically-acceptable salts thereof, and the MEK inhibitor is selected from the group consisting of trametinib, cobimetinib, and pharmaceutically-acceptable salts or solvates thereof 
     
     
         36 . The method of  claim 29 , wherein the one additional prior line of systemic therapy is a CTLA-4 inhibitor or a biosimilar thereof. 
     
     
         37 . The method of  claim 36 , wherein the CTLA-4 inhibitor is selected from the group consisting of ipilumumab, tremelimumab, and biosimilars thereof 
     
     
         38 . The method of  claim 29 , wherein the one additional prior line of systemic therapy is chemotherapeutic regimen. 
     
     
         39 . The method of  claim 38 , wherein the chemotherapeutic regimen comprises dacarbazine or temozolimide. 
     
     
         40 . The method of  claim 22 , wherein the first expansion is performed over a period of about 11 days. 
     
     
         41 . The method of any one of  claims 22 - 40 , wherein the IL-2 is present at an initial concentration of between 1000 IU/mL and 6000 IU/mL in the cell culture medium in the first expansion step (c). 
     
     
         42 . The method of any one of  claims 22 - 40 , wherein in the second expansion step (d), the IL-2 is present at an initial concentration of between 1000 IU/mL and 6000 IU/mL and the OKT-3 antibody is present at an initial concentration of about 30 ng/mL. 
     
     
         43 . The method of any one of  claims 22 - 42 , wherein the first expansion is performed using a gas permeable container. 
     
     
         44 . The method of any one of  claims 22 - 42 , wherein the second expansion is performed using a gas permeable container. 
     
     
         45 . The method of any one of  claims 22 - 44 , wherein the cell culture medium in the first expansion step (c) further comprises a cytokine selected from the group consisting of TL-4, TL-7, IL-15, IL-21, and combinations thereof. 
     
     
         46 . The method of any one of  claims 22 - 44 , wherein the cell culture medium in the second expansion step (d) further comprises a cytokine selected from the group consisting of IL-4, IL-7, IL-15, IL-21, and combinations thereof. 
     
     
         47 . The method of any one of  claims 22 - 46 , further comprising the step of treating the patient with a non-myeloablative lymphodepletion regimen prior to administering the TILs to the patient. 
     
     
         48 . The method of  claim 47 , wherein the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of 60 mg/m2/day for two days followed by administration of fludarabine at a dose of 25 mg/m2/day for five days. 
     
     
         49 . The method of any one of  claims 22 - 48 , further comprising the step of treating the patient with an IL-2 regimen starting on the day after administration of the TILs to the patient. 
     
     
         50 . The method of  claim 49 , wherein the IL-2 regimen is a high-dose IL-2 regimen comprising 600,000 or 720,000 11 J/kg of aldesleukin, or a biosimilar or variant thereof, administered as a 15-minute bolus intravenous infusion every eight hours until tolerance. 
     
     
         51 . The method according to any one of  claims 1 - 50 , wherein the therapeutically effective population of TILs comprises from about 2.3×10 10  to about 13.7×10 10  TILs.