Expansion culture of tumor-infiltrating lymphocytes (TILs) using adenosine A2A receptor antagonists, and therapeutic combinations of TILs and adenosine A2A receptor antagonists.

Abstract

To provide a method of expanding tumor infiltrating lymphocytes (TILs) to a therapeutic population of TILs.SOLUTION: Provided is a 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 to produce a second population of TILs; (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; (e) harvesting the therapeutic population of TILs obtained from step (d); and (f) transferring the harvested TIL population from step (e) to an infusion bag, the cell culture medium of step (c) and / or (d) comprising an adenosine 2A receptor antagonist.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Cross-reference of related applications

[0001] This international application claims priority to U.S. Provisional Patent Application No. 62 / 630,010 filed on 13 February 2018, U.S. Provisional Patent Application No. 62 / 637,603 filed on 2 March 2018, and U.S. Provisional Patent Application No. 62 / 684,698 filed on 13 June 2018, all of which are incorporated herein by reference.

[0002] Field of Invention

[0002] Disclosed herein are methods for expanding the culture of tumor-infiltrating lymphocytes (TILs) in the presence of adenosine A2A receptor (A2aR) antagonists such as vipardenant, siphoradenant (CPI-444), SCH58261, SYN115, ZM241385, SCH420814, xanthine superfamily A2aR antagonists, or related adenosine receptor 2A antagonists, as well as the use of expanded cultured TILs in the treatment of diseases such as cancer. Furthermore, therapeutic combinations of TILs and A2aR antagonists, including compositions and their use in the treatment of diseases such as cancer, are disclosed herein. [Background technology]

[0003]

[0003] Treatment of bulky, resistant cancers using adoptive autotransplantation of tumor-infiltrating lymphocytes (TILs) is a powerful approach to treating patients with poor prognoses. Gattinoni, et al., Nat. Rev. Immunol. 2006, 6, 383-393. TILs are T cell-dominant, and IL-2-based TIL expansion culture followed by a “rapid expansion culture process” (REP) has become the preferred method for TIL expansion culture due to 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. Several approaches have been explored to improve the clinical response to TIL therapy in melanoma and to extend TIL therapy to other types of tumors, but with limited success, this area remains challenging. Goff, et al., J. Clin. Oncol. 2016, 34, 2389-97; Dudley, et al., J. Clin. Oncol. 2008, 26, 5233-39; Rosenberg, et al., Clin. Cancer Res. 2011, 17, 4550-57. Specific subsets (CD8 +There has been considerable focus on the selection of TILs during expansion culture to select T cells (or other types of cells) or to target driver mutations such as the mutated ERBB2IP epitope or driver mutations in the KRAS oncogene. (Tran, et al., N. Engl.J. Med.2016, 375, 2255-62; Tran, et al., Science 2014, 344, 641-45). However, even if such selection approaches can be developed to demonstrate efficacy in large-scale clinical trials, they significantly increase the duration, complexity, and cost of TIL therapy, limiting the potential for widespread use of TIL therapy in different types of cancer.

[0004]

[0004] Adenosine A2A (or A 2A The receptors are A1, A 2B Adenosine is a member of the adenosine receptor group of G protein-coupled receptors, including A3, and is highly expressed in the spleen, thymus, leukocytes, platelets, and olfactory bulb. The presence of relatively high concentrations of adenosine in the immune microenvironment, which leads to A2a receptor activation, has been shown to represent a negative feedback loop that allows tumors to evade immune recognition. Therefore, A2A receptor (A2AR) antagonists are attracting attention as a novel form of checkpoint blockade for cancer immunotherapy. Leone, et al., Comp. Struct. Biotechnol. J. 2015, 13, 265-272. Immunosuppressive extracellular concentrations of adenosine in solid tumors are known to be in the μM range (10-20 times the normal concentration) and need to be overcome by A2AR antagonists. Blay, et al., Cancer Res. 1997, 57, 2602-2605. [Overview of the Initiative] [Means for solving the problem]

[0005]

[0005] The present invention provides the unexpected finding that adenosine receptor antagonists, such as A2AR antagonists, are useful for expanding the culture of TILs from tumors and are even more useful for treating patients in combination with TIL therapy.

[0006] Summary of the Invention

[0006] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and tumor necrosis factor receptor superfamily (TNFRSF) agonists, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to a patient with cancer. This provides a method that includes [something].

[0007]

[0007] A method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and an adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, peripheral blood mononuclear cells (PBMCs), and optionally an adenosine 2A receptor (A2aR) antagonist and a second adenosine 2A receptor (A2aR) antagonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to the patient. This method includes [something].

[0008]

[0008] In one embodiment, the present invention provides a method for expanding the culture of tumor-infiltrating lymphocytes (TILs).

[0009]

[0009] The present invention provides a method for expanding the culture of tumor-infiltrating lymphocytes (TILs), and this method is (a) Obtaining a tumor sample from a patient, wherein the tumor sample includes a first TIL population; (b) Processing the tumor sample into multiple tumor fragments; (c) Adding the tumor fragment to a sealed container; (d) Obtaining a second TIL population by performing an initial expansion culture of the first TIL population in a first cell culture medium, wherein the first cell culture medium comprises IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed in a medium of at least 100 cm². 2The initial expansion culture is carried out in a closed container that provides a gas-permeable surface area, and the initial expansion culture is carried out over a first period of about 7 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger in number than the first TIL population, and the transition from step (c) to step (d) occurs without opening the system; (e) Expanding the second TIL population in a second cell culture medium, the second cell culture medium comprising IL-2, OKT-3 and at least one adenosine 2A receptor (A2aR) antagonist and peripheral blood mononuclear cells (PBMCs, also known as mononuclear cells (MNCs)), wherein the expansion culture is carried out over a second period of about 7 to 14 days to obtain a third TIL population, the third TIL population exhibiting an increased effector T cell and / or central memory T cell subpopulation compared to the second TIL population, and the expansion culture is carried out over at least 500 cm². 2 The process is carried out in a closed container that provides a gas-permeable surface area, and the transition from step (d) to step (e) occurs without opening the system, during the expansion culture; (f) Recovering the third TIL group obtained from step (e), wherein the transition from step (e) to step (f) occurs without opening the system; and (g) (g) Transferring the recovered TIL mass from step (f) to an infusion bag, wherein the transfer from step (f) to (g) occurs without opening the system. Includes.

[0010]

[0010] In some embodiments, this method is an in vitro or ex vivo method.

[0011]

[0011] In some embodiments, the method further includes recovery via a cell processing system, such as a Fresenius Kabi LOVO system, in step (f). The term “LOVO cell processing system” also refers to any equipment or apparatus manufactured by any vendor that enables continuous flow and cell processing, in a sterile and / or closed system environment, through a membrane or filter such as a rotating membrane or rotating filter, to deliver a solution containing cells and remove the supernatant or cell culture without pelletizing. In some cases, the cell processing system can perform cell separation, washing, fluid exchange, concentration and / or other cell processing steps in a closed sterile system.

[0012]

[0012] In some embodiments, the closed container is selected from the group consisting of G containers and Xuri cell culture bags.

[0013]

[0013] In some embodiments, the infusion bag of step (g) is a HypoThermosol-containing infusion bag.

[0014]

[0014] In some embodiments, the first period of step (d) and the second period of step (e) are carried out individually within periods of 10, 11, or 12 days, respectively.

[0015]

[0015] In some embodiments, the first period of step (d) and the second period of step (e) are carried out separately within a period of 11 days.

[0016]

[0016] In some embodiments, steps (a) to (g) are carried out within a period of approximately 25 to 30 days.

[0017]

[0017] In some embodiments, steps (a) to (g) are carried out within a period of about 20 to 25 days.

[0018]

[0018] In some embodiments, steps (a) to (g) are carried out within a period of about 20 to 22 days.

[0019]

[0019] In some embodiments, steps (a) to (g) are carried out within 22 days.

[0020]

[0020] In some embodiments, steps (c) to (f) are performed in a single container, and performing steps (c) to (f) in a single container results in an increased TIL yield per excised tumor compared to performing steps (c) to (f) in multiple containers.

[0021]

[0021] In some embodiments, the PBMC is added to the TIL during the second period of step (e) without opening the system.

[0022]

[0022] In some embodiments, effector T cells and / or central memory T cells obtained from the third TIL population exhibit one or more features selected from the group consisting of CD27+ expression, CD28+ expression, longer telomeres, increased CD57 expression, and decreased CD56 expression, compared to effector T cells and / or central memory T cells obtained from the second cell population.

[0023]

[0023] In some embodiments, effector T cells and / or central memory T cells obtained from a third TIL population exhibit increased CD57 expression and decreased CD56 expression compared to effector T cells and / or central memory T cells obtained from a second cell population.

[0024]

[0024] In some embodiments, the risk of microbial contamination is reduced compared to open systems.

[0025]

[0025] In some embodiments, the TIL from step (g) is injected into the patient. In some embodiments, the TIL from step (g) is injected into the patient in combination with an adenosine A2A receptor antagonist. In some embodiments, the A2aR antagonist is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs thereof, and combinations thereof.

[0026]

[0026] The present invention also provides a method for treating a patient's cancer with a tumor-infiltrating lymphocyte (TIL) population, the method of which (a) A step of obtaining a tumor sample from a patient, wherein the tumor sample comprises a first TIL population; (b) The step of processing the tumor sample into multiple tumor fragments; (c) Adding the tumor fragment to a sealed container; (d) A step of obtaining a second TIL population by performing an initial expansion culture of the first TIL population in a first cell culture medium, wherein the first cell culture medium comprises IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, the initial expansion culture is performed in a closed container providing a gas-permeable surface area of ​​at least 100 cm2, the initial expansion culture is performed over a first period of about 7 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger in number than the first TIL population, and the transition from step (c) to step (d) occurs without opening the system; (e) Expanding the second TIL population in a second cell culture medium, the second cell culture medium comprising IL-2, OKT-3, and at least one adenosine 2A receptor (A2aR) antagonist and peripheral blood mononuclear cells (PBMCs), the expansion culture being carried out over a second period of about 7 to 14 days to obtain a third TIL population, the third TIL population exhibiting an increased effector T cell and / or central memory T cell subpopulation compared to the second TIL population, and the expansion culture being carried out over at least 500 cm². 2 The process is carried out in a closed container that provides a gas-permeable surface area, and the transition from step (d) to step (e) occurs without opening the system; (f) A step of recovering the third TIL group obtained from step (e), wherein the transition from step (e) to step (f) occurs without opening the system; (g) a step of transferring the recovered TIL mass from step (f) to an infusion bag, wherein the transfer from step (f) to (g) occurs without opening the system; and (h) Step (g) administering a therapeutically effective amount of TIL cells from the infusion bag of step (g) to the patient. Includes.

[0027]

[0027] In some embodiments, a therapeutically effective amount of TIL cells from the infusion bag in step (h) is administered to the patient in combination with an adenosine A2A receptor antagonist. In some embodiments, the A2aR antagonist is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof, or a combination thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs thereof, and combinations thereof.

[0028]

[0028] In some embodiments, the present invention also includes tumor-infiltrating lymphocyte populations for use in cancer treatment, the TIL population comprising: (b) processing a tumor sample obtained from a patient, the tumor sample comprising a first TIL population which is excised into a plurality of tumor fragments; (c) adding the tumor fragments to a closed container; (d) performing an initial expansion culture of the first TIL population in a first cell culture medium to obtain a second TIL population, the first cell culture medium comprising IL-2, and the initial expansion culture comprising at least 100 cm² 2Step (c) is carried out in a closed container that provides a gas-permeable surface area, and the initial expansion culture is carried out within a first period of about 7 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger in number than the first TIL population, and the transition from step (c) to step (d) occurs without opening the system; (e) expanding culture the second TIL population in a second cell culture medium, the second cell culture medium comprising IL-2, OKT-3 and at least one adenosine 2A receptor (A2aR) antagonist and peripheral blood mononuclear cells (PBMCs), the expansion culture is carried out within a second period of about 7 to 14 days to obtain a third TIL population, the third TIL population showing an increase in effector T cells and / or central memory T cell subpopulations compared to the second TIL population, and the expansion culture is carried out at least 500 cm² 2 The method is carried out in a closed container that provides a gas-permeable surface area, and the transition from step (d) to step (e) occurs without opening the system; (f) a step of recovering the third TIL population obtained from step (e), the transition from step (e) to step (f) occurs without opening the system; (g) a step of transferring the recovered TIL population from step (f) to an infusion bag, the transition from step (f) to (g) occurs without opening the system. In some embodiments, the method is obtained from a first step (a) of obtaining a tumor sample from a patient, the tumor sample comprising a first TIL population. In some embodiments, the TIL population is to be administered in a therapeutically effective dose from the infusion bag in step (g).

[0029]

[0029] In some embodiments, the third TIL population is maintained in a medium or formulation comprising an adenosine 2A receptor (A2aR) antagonist. In some embodiments, the A2aR antagonist is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal or prodrug thereof and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, bipadenant, ST4206, KF21213, SCH412348, 7MMG-49 or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal or prodrug thereof and combinations thereof.

[0030]

[0030] In some embodiments, a myeloablative lymphodepletion regimen is administered to the patient prior to administering a therapeutically effective amount of TIL cells in step (h). In some embodiments, the TIL population is for administration to a patient who has received a myeloablative lymphodepletion regimen.

[0031]

[0031] In some embodiments, the myeloablative lymphodepletion regimen comprises administration of cyclophosphamide at a dose of 60 mg / m 2 / day for 2 days, followed by administration of fludarabine at a dose of 25 mg / m 2 / day for 5 days.

[0032]

[0032] In some embodiments, the method further comprises treating the patient with a high-dose IL-2 regimen that begins on the day following administration of the TIL cells to the patient in step (h). In some embodiments, the TIL population is for administration prior to the high-dose IL-2 regimen. In some embodiments, the TIL population is for administration 1 day prior to the start of the high-dose IL-2 regimen.

[0033]

[0033] In some embodiments, the high-dose IL-2 regimen includes 600,000 or 720,000 IU / kg, which is administered as a 15-minute bolus intravenous infusion every 8 hours until an acceptable dose is reached.

[0034]

[0034] In some embodiments, effector T cells and / or central memory T cells obtained from the third TIL population exhibit one or more features selected from the group consisting of CD27+ expression, CD28+ expression, longer telomeres, increased CD57 expression, and decreased CD56 expression, compared to effector T cells and / or central memory T cells obtained from the second cell population.

[0035]

[0035] In some embodiments, effector T cells and / or central memory T cells obtained from a third TIL population exhibit increased CD57 expression and decreased CD56 expression compared to effector T cells and / or central memory T cells obtained from a second cell population.

[0036]

[0036] The present invention also provides a method for expanding tumor-infiltrating lymphocytes (TILs), the method comprising: (a) adding a treated tumor fragment to a closed system; (b) performing a first expansion culture of the first TIL population in a first cell culture medium to obtain a second TIL population, wherein the first cell culture medium comprises IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, the first expansion culture is performed in a closed container providing a first gas-permeable surface area, the first expansion culture is performed within a first period of about 3 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger in number than the first TIL population, and the transition from step (a) to step (b) occurring without opening the system; (c) expanding culture the second TIL population in a second cell culture medium, wherein the second cell culture medium comprises IL-2 The culture comprises OKT-3 and at least one adenosine 2A receptor (A2aR) antagonist and antigen-presenting cells, wherein the expansion culture is carried out over a second period of about 7 to 14 days to obtain a third TIL population, the third TIL population showing an increase in effector T cells and / or central memory T cell subpopulations compared to the second TIL population, the expansion culture is carried out in a closed container providing a second gas-permeable surface area, the transition from step (b) to step (c) occurs without opening the system, the step includes; (d) recovering the third TIL population obtained from step (c), the transition from step (c) to step (d) occurs without opening the system, and (e) transferring the recovered TIL population from step (d) to an infusion bag, the transition from step (d) to (e) occurs without opening the system.

[0037]

[0037] In some embodiments, the method further includes the step of cryopreserving the infusion bag containing the recovered TIL population using a cryopreservation process. In some embodiments, the cryopreservation process is carried out using a ratio of 1:1 between the recovered TIL population and CS10 medium.

[0038]

[0038] In some embodiments, the method further comprises adding an adenosine 2A receptor (A2aR) antagonist to a first TIL culture medium. In some embodiments, the A2aR antagonist is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof.

[0039]

[0039] In some embodiments, the method further comprises adding an adenosine 2A receptor (A2aR) antagonist to a second TIL culture medium. In some embodiments, the A2aR antagonist is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof.

[0040]

[0040] In some embodiments, the antigen-presenting cells are peripheral blood mononuclear cells (PBMCs). In some embodiments, the antigen-presenting cells are artificial antigen-presenting cells.

[0041]

[0041] In some embodiments, the collection in step (d) is carried out using a LOVO cell processing system.

[0042]

[0042] In some embodiments, the multiple fragments include about 50 fragments, each fragment being about 27 mm 3 It has a volume of approximately 1300 mm. In some embodiments, the multiple fragments are approximately 1300 mm 3 ~approx. 1500mm 3 It contains approximately 30 to 60 fragments with a total volume of approximately 1350 mm. In some embodiments, the fragments are approximately 1350 mm 3 It contains about 50 fragments having a total volume of . In some embodiments, the fragments include about 50 fragments having a total mass of about 1 g to about 1.5 g.

[0043]

[0043] In some embodiments, the second cell culture medium is provided in a container selected from the group consisting of G containers and Xuri cell culture bags.

[0044]

[0044] In some embodiments, the infusion bag of step (e) is a HypoThermosol-containing infusion bag.

[0045]

[0045] In some embodiments, the first period of step (b) and the second period of step (c) are carried out individually within a period of 10, 11, or 12 days, respectively. In some embodiments, the first period of step (b) and the second period of step (c) are carried out individually within a period of 11 days, respectively.

[0046]

[0046] In some embodiments, steps (a) to (e) are carried out within a period of about 25 to 30 days. In some embodiments, steps (a) to (e) are carried out within a period of about 20 to 25 days. In some embodiments, steps (a) to (e) are carried out within a period of about 20 to 22 days. In some embodiments, steps (a) to (e) are carried out within 22 days. In some embodiments, steps (a) to (e) and cryopreservation are carried out within 22 days.

[0047]

[0047] In some embodiments, steps (b) to (e) are performed in a single closed system, and performing steps (b) to (e) in a single container results in an increased TIL yield per excised tumor compared to performing steps (b) to (e) in multiple containers.

[0048]

[0048] In some embodiments, antigen-presenting cells are added to the TIL during the second period of step (c) without opening the system.

[0049]

[0049] In some embodiments, effector T cells and / or central memory T cells obtained from the third TIL population exhibit one or more features selected from the group consisting of CD27+ expression, CD28+ expression, longer telomeres, increased CD57 expression, and decreased CD56 expression, compared to effector T cells and / or central memory T cells obtained from the second cell population.

[0050]

[0050] In some embodiments, effector T cells and / or central memory T cells obtained from a third TIL population exhibit increased CD57 expression and decreased CD56 expression compared to effector T cells and / or central memory T cells obtained from a second cell population.

[0051]

[0051] In some embodiments, the risk of microbial contamination is reduced compared to open systems.

[0052]

[0052] In some embodiments, the TIL from step (e) is injected into the patient.

[0053]

[0053] In some embodiments, the TIL from step (e) is infused to the patient in combination with at least one adenosine 2A receptor antagonist. In some embodiments, the A2aR antagonist is CPI-444 or its pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or its pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs and combinations thereof.

[0054]

[0054] In some embodiments, the present invention also includes a tumor-infiltrating lymphocyte (TIL) population for use in the treatment of cancer, administered to patients receiving an adenosine 2A receptor antagonist (A2aR). In some embodiments, the A2aR is administered orally. In some embodiments, the A2aR is first co-administered with the tumor-infiltrating lymphocyte (TIL) population and then administered orally. In some embodiments, the A2aR is administered orally once daily. In some embodiments, the A2aR is administered orally twice daily. In some embodiments, the A2aR is administered orally three times daily. In some embodiments, the A2aR is CPI-444 or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal or prodrug thereof and combinations thereof. In some embodiments, the adenosine 2A receptor (A2aR) antagonist is selected from the group consisting of CPI-444, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, vipardenant, ST4206, KF21213, SCH412348, 7MMG-49 or pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs thereof, and combinations thereof.

[0055]

[0055] In some embodiments, the method further includes treating the patient with an adenosine 2A receptor antagonist (A2aR) before performing step (a). In some embodiments, the patient is treated for at least 1 day; 2 days; 3 days or more; 7 days or more; 7 days or more; less than 14 days; 14 days or more.

[0056]

[0056] In some embodiments, the closed container includes a single bioreactor. In some embodiments, the closed container includes a G-REX-10. In some embodiments, the closed container includes a G-REX-100. In some embodiments, the closed container includes a G-Rex 500. In some embodiments, the closed container includes a Xuri or Wave bioreactor gas permeable bag.

[0057]

[0057] In some embodiments, the Disclosure also provides a method for expanding the culture of tumor-infiltrating lymphocytes (TILs) into a therapeutic TIL population, the method being (b) Adding tumor fragments to a closed system, wherein the tumor fragments include a first TIL population; (c) Performing a first expansion culture to produce a second TIL population by culturing a first TIL population in a cell culture medium containing IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, wherein the first expansion culture is performed in a closed container providing a first gas-permeable surface area, the first expansion culture is performed for approximately 3 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger than the first TIL population, and the transition from step (b) to step (c) occurs without opening the system; (d) Performing a second expansion culture of the cell culture medium of the second TIL population by supplementing with additional IL-2, OKT-3, and at least one adenosine 2A receptor (A2aR) antagonist and antigen-presenting cells (APCs) to produce a third TIL population, the second expansion culture being performed over approximately 7–14 days to obtain the third TIL population, the third TIL population being a T-cell therapeutic TIL population including an increased subpopulation of effector T cells and / or central memory T cells compared to the second TIL population, the second expansion culture being performed in a closed vessel providing a second gas-permeable surface area, and the transition from step (c) to step (d) occurring without opening the system; (e) Recovering the therapeutic TIL population obtained from step (d), wherein the transition from step (d) to step (e) occurs without opening the system; and (f) Transferring the TIL population recovered from step (e) to an infusion bag, wherein the transition from step (e) to (f) occurs without opening the system. Includes.

[0058]

[0058] In some embodiments, the method includes, as a first step, (a) Obtain a first TIL population from tumors resected from patients by processing tumor samples obtained from patients into multiple tumor fragments. This also includes.

[0059]

[0059] In one embodiment, the method is an in vitro or ex vivo method.

[0060]

[0060] In some embodiments, the disclosure also provides a method for expanding the culture of tumor-infiltrating lymphocytes (TILs) into a therapeutic TIL population, the method being (a) Obtaining a first TIL population from tumors excised from patients by processing tumor samples obtained from patients into multiple tumor fragments; (b) Adding tumor fragments to a closed system; (c) Performing a first expansion culture to produce a second TIL population by culturing a first TIL population in a cell culture medium containing IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, wherein the first expansion culture is performed in a closed container providing a first gas-permeable surface area, the first expansion culture is performed for approximately 3 to 14 days to obtain a second TIL population, the second TIL population being at least 50 times larger than the first TIL population, and the transition from step (b) to step (c) occurs without opening the system; (d) Performing a second expansion culture of the cell culture medium of the second TIL population by supplementing it with additional IL-2, OKT-3 and optionally at least one adenosine 2A receptor (A2aR) antagonist and antigen-presenting cells (APCs) to produce a third TIL population, the second expansion culture being performed for approximately 7–14 days to obtain the third TIL population, the third TIL population being a T-cell therapeutic TIL population including an increased subpopulation of effector T cells and / or central memory T cells compared to the second TIL population, the second expansion culture being performed in a closed vessel providing a second gas-permeable surface area, and the transition from step (c) to step (d) occurring without opening the system; (e) Recovering the therapeutic TIL population obtained from step (d), wherein the transition from step (d) to step (e) occurs without opening the system; and (f) Transferring the TIL population recovered from step (e) to an infusion bag, wherein the transition from step (e) to (f) occurs without opening the system. Includes.

[0061]

[0061] In one embodiment, the method is an in vitro or ex vivo method.

[0062]

[0062] In some embodiments, the method further includes the step of freezing and preserving the infusion bag containing the TIL population recovered from step (f) using a cryopreservation process.

[0063]

[0063] In some embodiments, the cryopreservation process is carried out using a ratio of 1:1 between the recovered TIL population and the cryopreservation medium. In some embodiments, the cryopreservation medium contains dimethyl sulfoxide. In some embodiments, the cryopreservation medium is selected from the group consisting of Cryostor CS10, HypoThermosol, or a combination thereof.

[0064]

[0064] In some embodiments, the antigen-presenting cells are peripheral blood mononuclear cells (PBMCs).

[0065]

[0065] In some embodiments, the PBMCs are irradiated and homogeneous.

[0066]

[0066] In some embodiments, the PBMCs are added to the cell culture in step (d) on either day 9 to 14.

[0067]

[0067] In some embodiments, the antigen-presenting cells are artificial antigen-presenting cells.

[0068]

[0068] In some embodiments, the recovery in step (e) is carried out using the LOVO cell process system.

[0069]

[0069] In some embodiments, the tumor fragment is a plurality of fragments, comprising about 4 to about 50 fragments, each fragment being about 27 mm 3 It has a volume of approximately 1300 mm. In some embodiments, the multiple fragments are approximately 1300 mm 3 ~approx. 1500mm 3 It contains approximately 30 to 60 fragments with a total volume of approximately 1350 mm. In some embodiments, the fragments are approximately 1350 mm 3 It contains about 50 fragments having a total volume of . In some embodiments, the fragments include about 50 fragments having a total mass of about 1 g to about 1.5 g.

[0070]

[0070] In some embodiments, the cell culture medium is provided in a container selected from the group consisting of G containers and Xuri cell culture bags.

[0071]

[0071] In some embodiments, the infusion bag of step (f) is a HypoThermosol-containing infusion bag.

[0072]

[0072] In some embodiments, the first period of step (c) and the second period of step (e) are carried out individually within a period of 10, 11, or 12 days, respectively. In some embodiments, the first period of step (c) and the second period of step (e) are carried out individually within a period of 11 days, respectively. In some embodiments, steps (a) to (f) are carried out within a period of about 25 to 30 days. In some embodiments, steps (a) to (f) are carried out within a period of about 20 to 25 days. In some embodiments, steps (a) to (f) are carried out within a period of about 20 to 22 days. In some embodiments, steps (a) to (f) are carried out within 22 days. In some embodiments, steps (a) to (f) and cryopreservation are carried out within 22 days.

[0073]

[0073] In some embodiments, the therapeutic TIL population recovered from step (e) contains enough TILs to constitute a therapeutically effective dose of TILs. In some embodiments, the number of TILs sufficient to constitute a therapeutically effective dose is about 2.3 × 10¹⁰ to about 13.7 × 10¹⁰.

[0074]

[0074] In some embodiments, steps (b) to (e) are performed in a single container, and performing steps (b) to (e) in a single container results in an increased TIL yield per excised tumor compared to performing steps (b) to (e) in multiple containers.

[0075]

[0075] In some embodiments, antigen-presenting cells are added to the TIL during the second period of step (d) without opening the system.

[0076]

[0076] In some embodiments, effector T cells and / or central memory T cells in a therapeutic TIL population exhibit one or more features selected from the group consisting of CD27+ expression, CD28+ expression, longer telomeres, increased CD57 expression, and decreased CD56 expression, compared to effector T cells and / or central memory T cells obtained from a second cell population.

[0077]

[0077] In some embodiments, effector T cells and / or central memory T cells obtained from a third TIL population exhibit increased CD57 expression and decreased CD56 expression compared to effector T cells and / or central memory T cells obtained from a second cell population.

[0078]

[0078] In some embodiments, the risk of microbial contamination is reduced compared to open systems.

[0079]

[0079] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is selected from the group consisting of 4-1BB agonist, OX40 agonist, CD27 agonist, GITR agonist, HVEM agonist, CD95 agonist and combinations thereof. This provides a method that includes [something].

[0080]

[0080] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is selected from the group consisting of urelumab, utomirumab, EU-101 and its fragments, derivatives, variants, biosimilars and combinations thereof. This provides a method that includes [something].

[0081]

[0081] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium containing at least one adenosine 2A receptor (A2aR) antagonist, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is a 4-1BB agonist fusion protein. This provides a method that includes [something].

[0082]

[0082] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a 4-1BB agonist fusion protein, and the 4-1BB agonist fusion protein comprises (i) a first soluble 4-1BB binding domain, (ii) a first peptide linker, (iii) a second soluble 4-1BB binding domain, (iv) a second peptide linker, and (v) a third soluble 4-1BB binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB. This provides a method that includes [something].

[0083]

[0083] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an OX40 agonist, and the OX40 agonist is selected from the group consisting of tavorixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM-18455 and its fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0084]

[0084] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an OX40 agonist and the OX40 agonist is an OX40 agonist fusion protein. This provides a method that includes [something].

[0085]

[0085] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an OX40 agonist fusion protein, and the OX40 agonist fusion protein comprises (i) a first soluble OX40 binding domain, (ii) a first peptide linker, (iii) a second soluble OX40 binding domain, (iv) a second peptide linker, and (v) a third soluble OX40 binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB. This provides a method that includes [something].

[0086]

[0086] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a CD27 agonist, and the CD27 agonist is valrirumab or a fragment, derivative, variant or biosimilar thereof. This provides a method that includes [something].

[0087]

[0087] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a CD27 agonist and the CD27 agonist is a CD27 agonist fusion protein. This provides a method that includes [something].

[0088]

[0088] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a CD27 agonist and the CD27 agonist fusion protein comprises (i) a first soluble CD27 binding domain, (ii) a first peptide linker, (iii) a second soluble CD27 binding domain, (iv) a second peptide linker, and (v) a third soluble CD27 binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB. This provides a method that includes [something].

[0089]

[0089] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a GITR agonist, and the GITR agonists are TRX518, 6C8, 36E5, 3D6, 61G6, 6H6, 61F6, 1D8, 17F10, 35D8, 49A1, 9E5, 31H6, 2155, 698, 706, Steps selected from the group consisting of 827, 1649, 1718, 1D7, 33C9, 33F6, 34G4, 35B10, 41E11, 41G5, 42A11, 44C1, 45A8, 46E11, 48H12, 48H7, 49D9, 49E2, 48A9, 5H7, 7A10, 9H6 and their fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0090]

[0090] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a GITR agonist, and the GITR agonist is a GITR agonist fusion protein. This provides a method that includes [something].

[0091]

[0091] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is a GITR agonist fusion protein, and the GITR agonist fusion protein comprises (i) a first soluble GITR binding domain, (ii) a first peptide linker, (iii) a second soluble GITR binding domain, (iv) a second peptide linker, and (v) a third soluble GITR binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB. This provides a method that includes [something].

[0092]

[0092] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an HVEM agonist. This provides a method that includes [something].

[0093]

[0093] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an HVEM agonist, and the HVEM agonist is an HVEM agonist fusion protein. This provides a method that includes [something].

[0094]

[0094] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is an HVEM agonist fusion protein, and the HVEM agonist fusion protein comprises (i) a first soluble HVEM binding domain, (ii) a first peptide linker, (iii) a second soluble HVEM binding domain, (iv) a second peptide linker, and (v) a third soluble HVEM binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB. This provides a method that includes [something].

[0095]

[0095] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, further comprising a step of treating the patient with a TNFRSF agonist, which is administered intravenously at a dose of 0.1 mg / kg to 50 mg / kg every four weeks for up to eight cycles. This provides a method that includes [something].

[0096]

[0096] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a TNFRSF agonist prior to the step of resecting the tumor from the patient, wherein the TNFRSF agonist is administered intravenously at doses of 0.1 mg / kg to 50 mg / kg every four weeks for up to eight cycles. This provides a method that includes [something].

[0097]

[0097] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is selected from the group consisting of urelumab, utomirumab, EU-101, tavorixizumab, Creative Biolabs MOM-18455 and its fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0098]

[0098] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the first cell culture medium comprises a second TNFRSF agonist. This provides a method that includes [something].

[0099]

[0099] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is added to the first cell culture medium at intervals selected from the group consisting of daily, every two days, every three days, every four days, every five days, every six days, every seven days and every two weeks during the initial expansion culture. This provides a method that includes [something].

[0100]

[0100] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein a TNFRSF agonist and at least one adenosine 2A receptor (A2aR) antagonist are added to a second cell culture medium at intervals selected from the group consisting of daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every 7 days and every 2 weeks during rapid expansion culture. This provides a method that includes [something].

[0101]

[0101] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is added in a cell culture medium at a concentration sufficient to achieve a concentration of 0.1 μg / mL to 100 μg / mL, and at least one adenosine 2A receptor (A2aR) antagonist is added to achieve functional antagonism of the A2aR signaling pathway. This provides a method that includes [something].

[0102]

[0102] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the TNFRSF agonist is added to the cell culture medium at a concentration sufficient to achieve a concentration of 20 μg / mL to 40 μg / mL. This provides a method that includes [something].

[0103]

[0103] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the first cell culture medium at an initial concentration of about 10 to about 6000 IU / mL. This provides a method that includes [something].

[0104]

[0104] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the first cell culture medium at an initial concentration of approximately 3000 IU / mL. This provides a method that includes [something].

[0105]

[0105] In a further embodiment, the therapeutically active portion of a third TIL population is administered to a patient with cancer, and at least one adenosine 2A receptor (A2aR) antagonist is present in the first cell culture medium.

[0106]

[0106] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of a third TIL population to a patient having cancer (the method of claim 31), wherein IL-2 is present in a first cell culture medium at an initial concentration of about 800 IU / mL to about 1100 IU / mL. This provides a method that includes [something].

[0107]

[0107] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the first cell culture medium at an initial concentration of about 1000 IU / mL. This provides a method that includes [something].

[0108]

[0108] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the second cell culture medium at an initial concentration of about 10 to about 6000 IU / mL. This provides a method that includes [something].

[0109]

[0109] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the second cell culture medium at an initial concentration of approximately 3000 IU / mL. This provides a method that includes [something].

[0110]

[0110] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the second cell culture medium at an initial concentration of approximately 800 IU / mL to approximately 1100 IU / mL. This provides a method that includes [something].

[0111]

[0111] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-2 is present in the second cell culture medium at an initial concentration of approximately 1000 IU / mL, and the A2aR antagonist is present at a concentration sufficient to attenuate signaling via the A2aR pathway. This provides a method that includes [something].

[0112]

[0112] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-15 is present in the first cell culture medium. This provides a method that includes [something].

[0113]

[0113] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-15 is present in the first cell culture medium at an initial concentration of approximately 5 ng / mL to approximately 20 ng / mL. This provides a method that includes [something].

[0114]

[0114] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-15 is present in a second cell culture medium. This provides a method that includes [something].

[0115]

[0115] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-15 is present in the second cell culture medium at an initial concentration of approximately 5 ng / mL to approximately 20 ng / mL. This provides a method that includes [something].

[0116]

[0116] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-21 is present in the first cell culture medium. This provides a method that includes [something].

[0117]

[0117] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-21 is present in the first cell culture medium at an initial concentration of approximately 5 ng / mL to approximately 20 ng / mL. This provides a method that includes [something].

[0118]

[0118] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-21 is present in a second cell culture medium. This provides a method that includes [something].

[0119]

[0119] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein IL-21 is present in the second cell culture medium at an initial concentration of approximately 5 ng / mL to approximately 20 ng / mL. This provides a method that includes [something].

[0120]

[0120] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the OKT-3 antibody is present in the second cell culture medium at an initial concentration of approximately 10 ng / mL to approximately 60 ng / mL. This provides a method that includes [something].

[0121]

[0121] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the OKT-3 antibody is present in the second cell culture medium at an initial concentration of approximately 30 ng / mL. This provides a method that includes [something].

[0122]

[0122] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein the initial expansion culture is carried out using a gas-permeable container. This provides a method that includes [something].

[0123]

[0123] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, wherein rapid expansion culture is carried out using a gas-permeable container. This provides a method that includes [something].

[0124]

[0124] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a non-myeloablative lymphoid depletion regimen prior to administering the third TIL population to the patient. This provides a method that includes [something].

[0125]

[0125] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, further comprising the step of treating the patient with a non-myeloablative lymphoid depletion regimen prior to administering the third TIL population to the patient, the non-myeloablative lymphoid depletion regimen being 60 mg / m² 2 Administer cyclophosphamide at a daily dose for two days, followed by 25 mg / m². 2 The step includes administering fludarabine at a daily dose over a period of 5 days. This provides a method that includes [something].

[0126]

[0126] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, further comprising treating the patient with a tapering IL-2 regimen starting the day following the administration of the third TIL population to the patient, the tapering IL-2 regimen being 18,000,000 IU / m² on day 1. 2 On the second day, the reading was 9,000,000 IU / m³. 2 Furthermore, on the 3rd and 4th days, the levels were 4,500,000 IU / m³. 2 A step containing aldesleukin administered intravenously in the following doses. This provides a method that includes [something].

[0127]

[0127] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient with cancer, further comprising the step of administering the third TIL population to the patient at a dose of 0.10 mg / day to 50 mg / day, followed by treating the patient with pegylated IL-2. This provides a method that includes [something].

[0128]

[0128] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, further comprising the step of treating the patient with a high-dose IL-2 regimen starting the day following the administration of the third TIL population to the patient. This provides a method that includes [something].

[0129]

[0129] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient with cancer, further comprising treating the patient with a high-dose IL-2 regimen, which is initiated the day following the administration of the third TIL population to the patient, the high-dose IL-2 regimen comprising 600,000 or 720,000 IU / kg of aldesleukin or its biosimilar or variant, administered as a 15-minute bolus intravenous infusion every 8 hours up to a tolerable dose. This provides a method that includes [something].

[0130]

[0130] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, wherein the cancer is selected from the group consisting of melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer, renal cell carcinoma, acute myeloid leukemia, colorectal cancer, cholangiocarcinoma and sarcoma. This provides a method that includes [something].

[0131]

[0131] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC), triple-negative breast cancer, double-resistance melanoma and uveal (intraocular) melanoma. This provides a method that includes [something].

[0132]

[0132] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a PD-1 inhibitor or a PD-L1 inhibitor before surgically removing the tumor from the patient. This provides a method that includes [something].

[0133]

[0133] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a PD-1 inhibitor or a PD-L1 inhibitor before resecting the tumor from the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0134]

[0134] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with an adenosine 2a receptor (A2aR) antagonist after surgically removing a tumor from the patient. This provides a method that includes [something].

[0135]

[0135] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with (1) a PD-1 inhibitor or PD-L1 inhibitor and (2) an adenosine 2A receptor (A2aR) antagonist after surgically removing a tumor from the patient. This provides a method that includes [something].

[0136]

[0136] In one embodiment, the present invention relates to a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a PD-1 inhibitor or PD-L1 inhibitor after resecting the tumor from the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0137]

[0137] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutically effective portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a PD-1 inhibitor or a PD-L1 inhibitor after administering the third TIL population to the patient. This provides a method that includes [something].

[0138]

[0138] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs), and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; (e) the step of recovering the third TIL group; and (f) A step of administering the therapeutic active portion of a third TIL population to a patient having cancer, further comprising the step of treating the patient with a PD-1 inhibitor or a PD-L1 inhibitor after administering the third TIL population to the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations. This provides a method that includes [something].

[0139]

[0139] In one embodiment, the present invention is a process for preparing a tumor-infiltrating lymphocyte (TIL) population, (b) Step of obtaining the first TIL group; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; and (e) Step of recovering the third TIL group It provides a process that includes this.

[0140]

[0140] In one embodiment, the present invention is (b) Step of obtaining the first TIL group; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; and (e) Step of recovering the third TIL group This provides a population of tumor-infiltrating lymphocytes (TILs) that can be obtained from a process including the following.

[0141]

[0141] In one embodiment, the present invention provides a TIL population for use in the treatment of cancer. In one embodiment, the present invention provides a pharmaceutical composition comprising a tumor-infiltrating lymphocyte (TIL) population for use in the treatment of cancer, wherein the tumor-infiltrating lymphocyte (TIL) population is (b) Step of obtaining the first TIL group; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, at least one adenosine 2A receptor (A2aR) antagonist and a tumor necrosis factor receptor superfamily (TNFRSF) agonist, and the initial expansion culture is performed for a period of 21 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and optionally a TNFRSF agonist, and the rapid expansion culture is performed for a period of 14 days or less; and (e) Step of recovering the third TIL group It can be obtained through a process that includes this.

[0142]

[0142] In one embodiment, the first TIL population is obtained from a tumor. In one embodiment, the tumor is first excised from the patient. In one embodiment, the first TIL population is obtained from the tumor excised from the patient. In one embodiment, the TIL population is intended to be administered to a patient with cancer in a therapeutically effective dose.

[0143]

[0143] In one embodiment, the present invention is a method for expanding a population of tumor-infiltrating lymphocytes (TILs), (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2, and the initial expansion culture is performed for a period of 11 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and TNFRSF agonist, and the rapid expansion culture is performed for a period of 11 days or less; (e) the step of recovering the third TIL group; and (f) Optionally, cryopreserve a third TIL population in a dimethyl sulfoxide-based medium. This provides a method that includes [something].

[0144]

[0144] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed for a period of 11 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and TNFRSF agonist, and the rapid expansion culture is performed for a period of 11 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to the patient. This provides a method that includes [something].

[0145]

[0145] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed for a period of 11 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and TNFRSF agonist, and the rapid expansion culture is performed for a period of 11 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to the patient. The present invention provides a method in which the TNFRSF agonist is selected from the group consisting of 4-1BB agonists, OX40 agonists, and combinations thereof.

[0146]

[0146] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed for a period of 11 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and TNFRSF agonist, and the rapid expansion culture is performed for a period of 11 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to the patient. Includes, TNFRSF agonists are selected from the group consisting of 4-1BB agonists, OX40 agonists, and combinations thereof. The present invention provides a method in which a TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is selected from the group consisting of urelumab, utomirumab, EU-101, fusion proteins, and their fragments, derivatives, variants, biosimilars, and combinations.

[0147]

[0147] In one embodiment, the present invention is a method for treating cancer with a tumor-infiltrating lymphocyte (TIL) population, (a) The step of removing the tumor from the patient; (b) Steps to obtain a first TIL population from the tumor; (c) A step of obtaining a second TIL population by performing an initial expansion culture of a first TIL population in a first cell culture medium, wherein the second TIL population is at least five times larger in number than the first TIL population, the first cell culture medium contains IL-2 and at least one adenosine 2A receptor (A2aR) antagonist, and the initial expansion culture is performed for a period of 11 days or less; (d) A step of obtaining a third TIL population by performing rapid expansion culture of a second TIL population in a second cell culture medium, wherein seven days after the start of rapid expansion culture, the third TIL population is at least 50 times larger in number than the second TIL population, the second cell culture medium contains IL-2, OKT-3 (anti-CD3) antibody, at least one adenosine 2A receptor (A2aR) antagonist, peripheral blood mononuclear cells (PBMCs) and TNFRSF agonist, and the rapid expansion culture is performed for a period of 11 days or less; (e) the step of recovering the third TIL group; and (f) The step of administering the therapeutically effective portion of the third TIL population to the patient. Includes, TNFRSF agonists are selected from the group consisting of 4-1BB agonists, OX40 agonists, and combinations thereof. TNFRSF agonists are OX40 agonists, and OX40 agonists are selected from the group consisting of tavorixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM-18455, and their fragments, derivatives, variants, biosimilars, and combinations. The method provides a concentration of 1 μg / mL to 30 μg / mL of OX4 agonist at the start of step (d).

[0148]

[0148] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the TNFRSF agonist is present at a concentration of 5 μg / mL to 20 μg / mL at the start of step (d).

[0149]

[0149] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the TNFRSF agonist is present at a concentration of about 10 μg / mL at the start of step (d).

[0150]

[0150] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the TNFRSF agonist is maintained at a concentration of 1 μg / mL to 30 μg / mL throughout step (d).

[0151]

[0151] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the TNFRSF agonist is maintained at a concentration of 5 μg / mL to 20 μg / mL throughout step (d).

[0152]

[0152] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the TNFRSF agonist is maintained at a concentration of about 10 μg / mL throughout step (d).

[0153]

[0153] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein one adenosine 2A receptor (A2aR) antagonist is administered throughout step (d) at least 1 nM, about 10 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 85 nM, about 90 nM, about 95 nM, about 100 nM, about 1 uM, about 10 uM, about 25 uM M, approximately 50uM, approximately 75uM, approximately 80uM, approximately 90uM, approximately 100uM, approximately 125uM, approximately 150uM, approximately 175uM, approximately 200uM, approximately 225uM, approximately 250uM, approximately 280uM, approximately 275uM, approximately 290uM, approximately 300uM, less than 500uM, less than 1000uM, less than 2000uM, maintained at concentrations approximately equivalent to the solubility limit of a specific A2aR antagonist.

[0154]

[0154] In one embodiment, the present invention provides a method according to any of the embodiments described above, wherein the third TIL group is CD4 in the second TIL group. + CD8 for TIL + Compared to the TIL reference ratio, CD4 + CD8 for TIL + This shows the increased ratio of TIL. In one embodiment, the increased ratio is selected from the group consisting of at least 1% greater than the reference ratio, at least 2% greater than the reference ratio, at least 5% greater than the reference ratio, at least 10% greater than the reference ratio, at least 15% greater than the reference ratio, at least 20% greater than the reference ratio, at least 25% greater than the reference ratio, at least 30% greater than the reference ratio, at least 35% greater than the reference ratio, at least 40% greater than the reference ratio, at least 45% greater than the reference ratio, and at least 50% greater than the reference ratio. In one embodiment, the increased ratio is 5% to 80% greater than the reference ratio. In one embodiment, the increased ratio is 10% to 70% greater than the reference ratio. In one embodiment, the increased ratio is 15% to 60% greater than the reference ratio. In one of the embodiments described above, the reference ratio is obtained from a third TIL population that is a responder to the TNFRSF agonist.

[0155]

[0155] In one embodiment, the present invention provides a method of any of the embodiments described above, wherein the cancer is selected from the group consisting of melanoma, uveal (intraocular) melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer (head and neck squamous cell carcinoma), renal cell carcinoma, colorectal cancer, pancreatic cancer, glioblastoma, cholangiocarcinoma, and sarcoma. In one embodiment, the present invention provides a method of any of the embodiments described above, wherein the cancer is selected from the group consisting of cutaneous melanoma, uveal (intraocular) melanoma, platinum-resistant ovarian cancer, pancreatic ductal adenocarcinoma, osteosarcoma, triple-negative breast cancer, and non-small cell lung cancer.

[0156]

[0156] In one embodiment, any of the above embodiments may be combined with any of the following embodiments.

[0157]

[0157] In one embodiment, the process is an in vitro or ex vivo process.

[0158]

[0158] In one embodiment, the TNFRSF agonist is selected from the group consisting of 4-1BB agonist, OX40 agonist, CD27 agonist, GITR agonist, HVEM agonist, CD95 agonist, and combinations thereof.

[0159]

[0159] In one embodiment, the TNFRSF agonist is a 4-1BB agonist.

[0160]

[0160] In one embodiment, the TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is selected from the group consisting of urelumab, utomirumab, EU-101, and their fragments, derivatives, variants, biosimilars, and combinations.

[0161]

[0161] In one embodiment, the TNFRSF agonist is a 4-1BB agonist, and the 4-1BB agonist is a 4-1BB agonist fusion protein.

[0162]

[0162] In one embodiment, the TNFRSF agonist is a 4-1BB agonist fusion protein, and the 4-1BB agonist fusion protein comprises (i) a first soluble 4-1BB binding domain, (ii) a first peptide linker, (iii) a second soluble 4-1BB binding domain, (iv) a second peptide linker, and (v) a third soluble 4-1BB binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB.

[0163]

[0163] In one embodiment, the TNFRSF agonist is an OX40 agonist.

[0164]

[0164] In one embodiment, the TNFRSF agonist is an OX40 agonist, and the OX40 agonist is selected from the group consisting of tavorixizumab, GSK3174998, MEDI6469, MEDI6383, MOXR0916, PF-04518600, Creative Biolabs MOM-18455, and their fragments, derivatives, variants, biosimilars and combinations.

[0165]

[0165] In one embodiment, the TNFRSF agonist is an OX40 agonist, and the OX40 agonist is an OX40 agonist fusion protein.

[0166]

[0166] In one embodiment, the TNFRSF agonist is an OX40 agonist fusion protein, and the OX40 agonist fusion protein comprises (i) a first soluble OX40 binding domain, (ii) a first peptide linker, (iii) a second soluble OX40 binding domain, (iv) a second peptide linker, and (v) a third soluble OX40 binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB.

[0167]

[0167] In one embodiment, the TNFRSF agonist is a CD27 agonist.

[0168]

[0168] In one embodiment, the TNFRSF agonist is a CD27 agonist, and the CD27 agonist is valrirumab or a fragment, derivative, variant, or biosimilar thereof.

[0169]

[0169] In one embodiment, the TNFRSF agonist is a CD27 agonist, and the CD27 agonist is a CD27 agonist fusion protein.

[0170]

[0170] In one embodiment, the TNFRSF agonist is a CD27 agonist, and the CD27 agonist fusion protein comprises (i) a first soluble CD27 binding domain, (ii) a first peptide linker, (iii) a second soluble CD27 binding domain, (iv) a second peptide linker, and (v) a third soluble CD27 binding domain, further comprising additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB.

[0171]

[0171] In one embodiment, the TNFRSF agonist is a GITR agonist.

[0172]

[0172] In one embodiment, the TNFRSF agonist is a GITR agonist, and the GITR agonist is selected from the group consisting of TRX518, 6C8, 36E5, 3D6, 61G6, 6H6, 61F6, 1D8, 17F10, 35D8, 49A1, 9E5, 31H6, 2155, 698, 706, 827, 1649, 1718, 1D7, 33C9, 33F6, 34G4, 35B10, 41E11, 41G5, 42A11, 44C1, 45A8, 46E11, 48H12, 48H7, 49D9, 49E2, 48A9, 5H7, 7A10, 9H6 and their fragments, derivatives, variants, biosimilars and combinations.

[0173]

[0173] In one embodiment, the TNFRSF agonist is a GITR agonist, and the GITR agonist is a GITR agonist fusion protein.

[0174]

[0174] In one embodiment, the TNFRSF agonist is a GITR agonist fusion protein, and the GITR agonist fusion protein comprises (i) a first soluble GITR binding domain, (ii) a first peptide linker, (iii) a second soluble GITR binding domain, (iv) a second peptide linker, and (v) a third soluble GITR binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB.

[0175]

[0175] In one embodiment, the TNFRSF agonist is an HVEM agonist.

[0176]

[0176] In one embodiment, the TNFRSF agonist is an HVEM agonist, and the HVEM agonist is an HVEM agonist fusion protein.

[0177]

[0177] In one embodiment, the TNFRSF agonist is an HVEM agonist fusion protein, and the HVEM agonist fusion protein comprises (i) a first soluble HVEM binding domain, (ii) a first peptide linker, (iii) a second soluble HVEM binding domain, (iv) a second peptide linker, and (v) a third soluble HVEM binding domain, and further comprises additional domains at the N-terminus and / or C-terminus, the additional domains comprising an Fc fragment domain and a hinge domain, and the fusion protein has a dimeric structure according to structure IA or structure IB.

[0178]

[0178] In one embodiment, the TNFRSF agonist is selected from the group consisting of urelumab, utomirumab, EU-101, tavorixizumab, Creative Biolabs MOM-18455, and their fragments, derivatives, variants, biosimilars, and combinations.

[0179]

[0179] In one embodiment, the first cell culture medium contains a second TNFRSF agonist.

[0180]

[0180] In one embodiment, the TNFRSF agonist is added to the first cell culture medium at intervals selected from the group consisting of daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every 7 days and every 2 weeks during the initial expansion culture.

[0181]

[0181] In one embodiment, the TNFRSF agonist is added to the second cell culture medium at intervals selected from the group consisting of daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every 7 days and every 2 weeks during rapid expansion culture.

[0182]

[0182] In one embodiment, the TNFRSF agonist is added to the cell culture medium at a concentration sufficient to achieve a concentration of 0.1 μg / mL to 100 μg / mL.

[0183]

[0183] In one embodiment, the TNFRSF agonist is added to the cell culture medium at a concentration sufficient to achieve a concentration of 20 μg / mL to 40 μg / mL.

[0184]

[0184] Further details of the TNFRSF agonist are provided herein.

[0185]

[0185] In one embodiment, IL-2 is present in the first cell culture medium at an initial concentration of about 10 to about 6000 IU / mL.

[0186]

[0186] In one embodiment, IL-2 is present in the first cell culture medium at an initial concentration of about 3000 IU / mL.

[0187]

[0187] In one embodiment, IL-2 is present in the first cell culture medium at an initial concentration of about 800 IU / mL to about 1100 IU / mL.

[0188]

[0188] In one embodiment, IL-2 is present in the first cell culture medium at an initial concentration of about 1000 IU / mL.

[0189]

[0189] In one embodiment, IL-2 is present in the second cell culture medium at an initial concentration of about 10 to about 6000 IU / mL.

[0190]

[0190] In one embodiment, IL-2 is present in the second cell culture medium at an initial concentration of about 3000 IU / mL.

[0191]

[0191] In one embodiment, IL-2 is present in the second cell culture medium at an initial concentration of about 800 IU / mL to about 1100 IU / mL.

[0192]

[0192] In one embodiment, IL-2 is present in the second cell culture medium at an initial concentration of about 1000 IU / mL.

[0193]

[0193] In one embodiment, IL-15 is present in the first cell culture medium.

[0194]

[0194] In one embodiment, IL-15 is present in the first cell culture medium at an initial concentration of about 5 ng / mL to about 20 ng / mL.

[0195]

[0195] In one embodiment, IL-15 is present in a second cell culture medium.

[0196]

[0196] In one embodiment, IL-15 is present in the second cell culture medium at an initial concentration of about 5 ng / mL to about 20 ng / mL.

[0197]

[0197] In one embodiment, IL-21 is present in a first cell culture medium.

[0198]

[0198] In one embodiment, IL-21 is present in the first cell culture medium at an initial concentration of about 5 ng / mL to about 20 ng / mL.

[0199]

[0199] In one embodiment, IL-21 is present in a second cell culture medium.

[0200]

[0200] In one embodiment, IL-21 is present in the second cell culture medium at an initial concentration of about 5 ng / mL to about 20 ng / mL.

[0201]

[0201] In one embodiment, the OKT-3 antibody is present in the second cell culture medium at an initial concentration of about 10 ng / mL to about 60 ng / mL.

[0202]

[0202] In one embodiment, the OKT-3 antibody is present in the second cell culture medium at an initial concentration of approximately 30 ng / mL.

[0203]

[0203] In one embodiment, the initial expansion culture is carried out using a gas-permeable container.

[0204]

[0204] In one embodiment, rapid expansion culture is carried out using a gas-permeable container.

[0205]

[0205] In one embodiment, the present invention provides a tumor-infiltrating lymphocyte (TIL) population for use in the treatment of cancer, wherein the tumor-infiltrating lymphocyte (TIL) population can be obtained by a process of the present invention as described herein.

[0206]

[0206] In one embodiment, the present invention provides a pharmaceutical composition comprising a population of tumor-infiltrating lymphocytes (TILs) for use in a method of treating cancer, wherein the population of tumor-infiltrating lymphocytes (TILs) can be obtained by a process of the present invention as described herein.

[0207]

[0207] In one embodiment, the TIL group and / or pharmaceutical composition is for use in the treatment of cancer in combination with TNFRSF.

[0208]

[0208] In one embodiment, the present invention provides a combination of a TIL population that can be obtained by the process of the present invention as described herein and TNFRSF for use in the treatment of cancer.

[0209]

[0209] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a TNFRSF agonist, wherein the TNFRSF agonist is to be administered to the patient the day following the administration of a third TIL population, and the TNFRSF agonist is administered intravenously at doses of 0.1 mg / kg to 50 mg / kg every four weeks for up to eight cycles.

[0210]

[0210] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a TNFRSF agonist, wherein the TNFRSF agonist is to be administered before the step of tumor resection from the patient, and the TNFRSF agonist is to be administered intravenously at doses of 0.1 mg / kg to 50 mg / kg every 4 weeks for up to 8 cycles.

[0211]

[0211] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a non-myeloablative lymphoid depletion regimen.

[0212]

[0212] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a non-myeloablative lymphoid depletion regime before administering to the patient the pharmaceutical composition comprising the third TIL population and / or the third TIL population.

[0213]

[0213] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a non-myeloablative lymph depletion regimen prior to administration of the pharmaceutical composition comprising the third TIL population and / or the third TIL population to the patient, wherein the non-myeloablative lymph depletion regimen is 60 mg / m² 2 Administer cyclophosphamide at a daily dose for two days, followed by 25 mg / m². 2 The procedure includes administering fludarabine at a dose of / day over a period of 5 days. Further details of the non-myeloablative lymphoid depletion regimen are provided herein, for example, under the heading "Non-myeloablative lymphoid depletion by chemotherapy."

[0214]

[0214] In one embodiment, the TIL group and / or pharmaceutical composition is intended for use in the treatment of cancer in combination with an IL-2 regimen.

[0215]

[0215] In one embodiment, the IL-2 regimen is a tapering IL-2 regimen.

[0216]

[0216] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a tapering IL-2 regimen initiated the day following administration of the third TIL population and / or pharmaceutical composition containing the third TIL population to the patient, wherein the tapering IL-2 regimen is 18,000,000 IU / m² on day 1. 2 On the second day, the reading was 9,000,000 IU / m³. 2 Furthermore, on the 3rd and 4th days, the levels were 4,500,000 IU / m³. 2 Contains aldezleukin administered intravenously at the specified dose.

[0217]

[0217] In one embodiment, the TIL group and / or pharmaceutical composition is for use in the treatment of cancer in combination with pegylated IL-2.

[0218]

[0218] In one embodiment, the TIL population and / or pharmaceutical composition is for use in a method of treating cancer in combination with pegylated IL-2, which is administered to a patient after administering a pharmaceutical composition containing a third TIL population and / or a third TIL population at a dose of 0.10 mg / day to 50 mg / day.

[0219]

[0219] In one embodiment, the TIL population and / or pharmaceutical composition is for use in a method of treating cancer in combination with a high-dose IL-2 regimen.

[0220]

[0220] In one embodiment, the TIL population and / or pharmaceutical composition is for use in a method of treating cancer in combination with a high-dose IL-2 regimen initiated the day after the administration of the pharmaceutical composition containing the third TIL population and / or the third TIL population to the patient.

[0221]

[0221] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a high-dose IL-2 regimen initiated the day following administration of a third TIL population and / or a pharmaceutical composition comprising the third TIL population to a patient, wherein the high-dose IL-2 regimen comprises 600,000 or 720,000 IU / kg of aldesleukin or its biosimilar or variant, administered as a 15-minute bolus intravenous infusion every 8 hours up to a tolerable dose.

[0222]

[0222] In one embodiment, the TIL group and / or pharmaceutical composition is for use in the treatment of cancer, the cancer being selected from the group consisting of melanoma, ovarian cancer, cervical cancer, lung cancer, bladder cancer, breast cancer, head and neck cancer, renal cell carcinoma, acute myeloid leukemia, colorectal cancer, cholangiocarcinoma and sarcoma.

[0223]

[0223] In one embodiment, the TIL group and / or pharmaceutical composition is for use in the treatment of cancer, the cancer being selected from the group consisting of non-small cell lung cancer (NSCLC), triple-negative breast cancer, double-resistance melanoma and uveal (intraocular) melanoma.

[0224]

[0224] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor.

[0225]

[0225] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations.

[0226]

[0226] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor, wherein the PD-1 inhibitor or PD-L1 inhibitor is to be administered before the tumor is removed from the patient.

[0227]

[0227] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or PD-L1 inhibitor before surgical removal of the tumor from the patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations.

[0228]

[0228] In one embodiment, the TIL population and / or pharmaceutical composition is for use in a method of treating cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor.

[0229]

[0229] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations.

[0230]

[0230] In one embodiment, the TIL population and / or pharmaceutical composition is for use in a method of treating cancer in combination with a PD-1 inhibitor or PD-L1 inhibitor after tumor resection from a patient.

[0231]

[0231] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or PD-L1 inhibitor after tumor resection from a patient, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations.

[0232]

[0232] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or a PD-L1 inhibitor, wherein the PD-1 or PD-L1 inhibitor is for administration to the patient after administration of a pharmaceutical composition containing a third TIL population and / or a third TIL population.

[0233]

[0233] In one embodiment, the TIL population and / or pharmaceutical composition is for use in the treatment of cancer in combination with a PD-1 inhibitor or PD-L1 inhibitor for administration after the patient has been administered a third TIL population, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab and their fragments, derivatives, variants, biosimilars and combinations. Further details of the PD-1 inhibitor and PD-L1 inhibitor are described herein, for example, under the heading "Combinations with PD-1 and PD-L1 Inhibitors". In some embodiments, the TIL population and / or pharmaceutical composition containing the TIL population further includes one or more features as described herein, for example, under the headings "Pharmaceutical Compositions, Dosages and Dosage Regimen of TILs" and "Pharmaceutical Compositions, Dosages and Dosage Regimen of TNFRSF Agonists".

[0234] Brief explanation of the drawing

[0234] The above summary and the following detailed description of the invention will be better understood when read in conjunction with the attached drawings. [Brief explanation of the drawing]

[0235] [Figure 1]

[0235] The TIL expansion culture and treatment process is illustrated. The A2AR antagonist (labeled "A2AR" in Figure 1) or TNFRSF agonist of the present invention can be used in both the pre-REP stage (upper half of the figure) or the REP stage (lower half of the figure) and can be added when IL-2 is added to each cell culture. Step 1 refers to the addition of approximately 4 tumor fragments to 10 G-Rex 10 flasks. In Step 2, approximately 40 × 10⁶ TILs or more are obtained. In Step 3, the fragmentation into 36 G-Rex 100 flasks occurs for REP. In Step 4, the TILs are recovered by centrifugation. The fresh TIL product is obtained in Step 5 after a total process time of approximately 43 days, at which point the TILs can be injected into the patient. [Figure 2]

[0236] This disclosure provides a treatment protocol for use with TILs grown with the A2AR antagonist. The TNFRSF agonist may be used in treatment as described herein, either after administration of TILs or during the expansion culture process. [Figure 3]

[0237] An exemplary TIL expansion culture and manufacturing protocol (Process 2A) is shown. [Figure 4]

[0238] The steps of an exemplary method performed in Process 2A are shown below. [Figure 5]

[0239] An exemplary TIL expansion culture protocol is shown. [Figure 6]

[0240] This study evaluates the binding affinity to Creative Biolabs (CB) and BPS Biosciences (BPS) 4-1BB agonist antibodies, as assessed by the percentage of 4-1BB+ cells obtained by flow cytometry. The CB 4-1BB agonist showed the highest binding affinity. [Figure 7]

[0241] This shows the binding affinity to Creative Biolabs (CB) and BPS Biosciences (BPS) 4-1BB agonist antibodies, evaluated by mean fluorescence intensity (MFI). The CB 4-1BB agonist showed the highest binding affinity. [Figure 8]

[0242] The results of the evaluation of NF-κB pathway activation by anti-4-1BB agonist antibodies are shown. [Figure 9]

[0243] This indicates the binding affinity to the Creative Biolabs OX40 agonist antibody, as assessed by the percentage of OX40+ cells obtained by flow cytometry. [Figure 10]

[0244] This indicates the binding affinity to the Creative Biolabs OX40 agonist antibody, as evaluated by mean fluorescence intensity (MFI). [Figure 11]

[0245] Creative Biolabs anti-OX40 agonist antibody (at the five concentrations shown) exhibits equivalent binding affinity to the commercially available anti-OX40 (clone Ber-ACT35) agonist. The first letter of each tumor name indicates histology: C=cervix; H=head and neck (head and neck squamous cell carcinoma); L=lung; and M=melanoma. [Figure 12]

[0246] The results of the evaluation of NF-κB pathway activation by anti-OX40 agonist antibodies are presented. OX40 reporter cells were treated for 24 hours with either anti-OX40 alone or an isotype control at concentrations of 1, 2, 4, 8, and 16 μg / mL, with or without PBMC feeder cells. The cells were lysed using a one-step luciferase reagent, and luciferase activity was measured using a luminometer. [Figure 13]

[0247] This document presents the experimental design for experiments using 4-1BB and OX40 agonists during the pre-REP phase. [Figure 14]

[0248] The tumor histology used in the experimental design shown in Figure 23 is presented. [Figure 15]

[0249] This document presents a data analysis strategy used to evaluate the impact of 4-1BB and anti-OX40 agonists used during pre-REP on TIL performance and characteristics. [Figure 16]

[0250] The results for total cell count in cell expansion cultures using CB 4-1BB agonist are shown (N=3). NT = no test (control). The p-value was >0.99. [Figure 17]

[0251] The results for total cell count in cell expansion cultures using CB OX40 agonist are shown (N=5). NT=not tested (control). The p-value was 0.06. [Figure 18]

[0252] The results for total cell count in cell expansion cultures using CB 4-1BB agonist and OX-40 agonist are shown (N=2). NT = No test (control). [Figure 19]

[0253] The results for the total CD8+ cell count in cell expansion cultures using CB 4-1BB agonist are shown (N=3). The p-value was 0.5. [Figure 20]

[0254] The results for total CD8+ cell count in cell expansion cultures using CB OX40 agonist are shown (N=5). The p-value was 0.03. [Figure 21]

[0255] The results for total CD8+ cell counts in cell expansion cultures using CB 4-1BB agonist and OX-40 agonist are shown (N=2). NT = not tested (control). [Figure 22]

[0256] The results for the total CD8+ / CD4+ cell count ratio in cell expansion cultures using CB 4-1BB agonist are shown (N=3). The p-value was 0.2. [Figure 23]

[0257] The results for the total CD8+ / CD4+ cell count ratio in cell expansion cultures using CB OX40 agonist are shown (N=5). The p-value was 0.12. [Figure 24]

[0258] The results for the total CD8+ / CD4+ cell count ratio in cell expansion cultures using CB 4-1BB agonist and OX-40 agonist are shown (N=2). NT = not tested (control). [Figure 25]

[0259] The experimental scheme for REP proliferation of pre-REP TILs cultured in the presence of 4-1BB or OX40 agonist is shown. [Figure 26]

[0260] This shows the magnification factor of pre-REP TILs cultured in the presence of CB 4-1BB agonist compared to TILs not treated with pre-REP(NT), and TILs cultured with REP. [Figure 27]

[0261] This shows the magnification factor of pre-REP TILs cultured in the presence of CB OX40 agonist compared to TILs not treated with pre-REP(NT), and TILs cultured with REP. [Figure 28]

[0262] This shows the magnification factor of pre-REP TILs cultured in the presence of CB 4-1BB agonist and CB OX40 agonist compared to TILs not treated with pre-REP(NT), and TILs cultured with REP. [Figure 29]

[0263] Histology of 21 TIL strains used for evaluation of CB OX40 agonists during the REP phase is shown. [Figure 30]

[0264] This document presents an experimental scheme for evaluating CB OX40 agonists during the REP phase. [Figure 31]

[0265] The presence of the OX40 agonist antibody indicates that CD8+ TILs are preferentially expanded during REP (shown as a percentage of CD3+CD4+ cells). [Figure 32]

[0266] The presence of the OX40 agonist antibody indicates that CD8+ TILs are preferentially expanded during REP (shown as a percentage of CD3+CD8+ cells). [Figure 33]

[0267] This indicates that in non-responder TIL lines, no downregulation of OX40 was observed in the CD4+ subset after anti-OX40 treatment. [Figure 34]

[0268] This document presents experimental details of CB OX40 agonist dose titration in non-responder and responder TIL systems. [Figure 35]

[0269] The results of CB OX40 agonist dose titration in the Responder TIL system are shown. [Figure 36]

[0270] The results of CB OX40 agonist dose titration in non-responder TIL systems are shown. [Figure 37]

[0271] This shows a comparable TCRvb repertoire profile for the responder L4005. [Figure 38]

[0272] This shows a comparable TCRvb repertoire profile for responder H3005. [Figure 39]

[0273] This shows a comparable TCRvb repertoire profile for the M1022 responder. [Figure 40]

[0274] The cell count results for melanoma TILs obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 41]

[0275] The cell count results for lung TILs (first tumors) obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 42]

[0276] The cell count results for lung TILs (secondary tumors) obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 43]

[0277] Flow cytometry analysis of CD8+ and CD4+ subsets of melanoma TILs obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions is shown. [Figure 44]

[0278] Flow cytometry analysis of CD8+ and CD4+ subsets of pulmonary TILs (primary tumors) obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions is shown. [Figure 45]

[0279] Flow cytometry analysis of CD8+ and CD4+ subsets of pulmonary TILs (secondary tumors) obtained after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions is shown. [Figure 46]

[0280] The results of ELISA and ELIspot obtained from melanoma TILs after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 47]

[0281] The results of ELISA and ELIspot obtained from lung TILs (first tumors) after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 48]

[0282] The results of ELISA and ELIspot obtained from lung TILs (secondary tumors) after the addition of A2AR antagonists to pre-REP and REP cultures under various conditions are shown. [Figure 49]

[0283] This disclosure provides a treatment protocol for use with TILs grown with the A2AR antagonist. The TNFRSF agonist may be used in treatment as described herein, either after administration of TILs or during the expansion culture process. [Modes for carrying out the invention]

[0236] A brief explanation of sequence listings

[0284] Sequence ID 1 is the amino acid sequence of the heavy chain of muromonab.

[0285] Sequence ID 2 is the amino acid sequence of the light chain of muromonab.

[0286] Sequence ID 3 is the amino acid sequence of recombinant human IL-2 protein.

[0287] Sequence ID 4 is the amino acid sequence of aldethleukin.

[0288] Sequence ID 5 is the amino acid sequence of recombinant human IL-4 protein.

[0289] Sequence ID 6 is the amino acid sequence of recombinant human IL-7 protein.

[0290] Sequence ID 7 is the amino acid sequence of recombinant human IL-15 protein.

[0291] Sequence ID 8 is the amino acid sequence of recombinant human IL-21 protein.

[0292] Sequence ID 9 is the amino acid sequence of human 4-1BB.

[0293] Sequence ID 10 is the amino acid sequence of mouse 4-1BB.

[0294] Sequence ID 11 is the heavy chain of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0295] Sequence ID 12 is the light chain of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0296] Sequence ID 13 is the heavy chain variable region (VH) of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0297] Sequence ID 14 is the light chain variable region (VL) of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0298] Sequence ID 15 is the heavy chain CDR1 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0299] Sequence ID 16 is the heavy chain CDR2 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0300] Sequence ID No. 17 is the heavy chain CDR3 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0301] Sequence ID No. 18 is the light chain CDR1 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0302] Sequence ID 19 is the light chain CDR2 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0303] Sequence ID No. 20 is the light chain CDR3 of the 4-1BB agonist monoclonal antibody utomirumab (PF-05082566).

[0304] Sequence ID 21 is the heavy chain of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0305] Sequence ID 22 is the light chain of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0306] Sequence ID 23 is the heavy chain variable region (VH) of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0307] Sequence ID No. 24 is the light chain variable region (VL) of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0308] Sequence ID No. 25 is the heavy chain CDR1 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0309] Sequence ID No. 26 is the heavy chain CDR2 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0310] Sequence ID No. 27 is the heavy chain CDR3 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0311] Sequence ID No. 28 is the light chain CDR1 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0312] Sequence ID No. 29 is the light chain CDR2 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0313] Sequence ID No. 30 is the light chain CDR3 of the 4-1BB agonist monoclonal antibody urelumab (BMS-663513).

[0314] Sequence ID 31 is the Fc domain of the TNFRSF agonist fusion protein.

[0315] Sequence ID 32 is the linker for the TNFRSF agonist fusion protein.

[0316] Sequence ID 33 is the linker for the TNFRSF agonist fusion protein.

[0317] Sequence ID 34 is the linker for the TNFRSF agonist fusion protein.

[0318] Sequence ID 35 is the linker for the TNFRSF agonist fusion protein.

[0319] Sequence ID 36 is the linker for the TNFRSF agonist fusion protein.

[0320] Sequence ID 37 is the linker for the TNFRSF agonist fusion protein.

[0321] Sequence ID 38 is the linker for the TNFRSF agonist fusion protein.

[0322] Sequence ID 39 is the linker for the TNFRSF agonist fusion protein.

[0323] Sequence ID 40 is the linker for the TNFRSF agonist fusion protein.

[0324] Sequence ID 41 is the linker for the TNFRSF agonist fusion protein.

[0325] Sequence ID 42 is the Fc domain of the TNFRSF agonist fusion protein.

[0326] Sequence ID 43 is the linker for the TNFRSF agonist fusion protein.

[0327] Sequence ID 44 is the linker for the TNFRSF agonist fusion protein.

[0328] Sequence ID 45 is the linker for the TNFRSF agonist fusion protein.

[0329] Sequence ID 46 is the amino acid sequence of the 4-1BB ligand (4-1BBL).

[0330] Sequence ID No. 47 is the soluble portion of the 4-1BBL polypeptide.

[0331] Sequence ID 48 is the heavy chain variable region (VH) of the 4-1BB agonist antibody 4B4-1-1 version 1.

[0332] Sequence ID 49 is the light chain variable region (VL) of the 4-1BB agonist antibody 4B4-1-1 version 1.

[0333] Sequence ID 50 is the heavy chain variable region (VH) of the 4-1BB agonist antibody 4B4-1-1 version 2.

[0334] Sequence ID 51 is the light chain variable region (VL) of the 4-1BB agonist antibody 4B4-1-1 version 2.

[0335] Sequence ID 52 is the heavy chain variable region (VH) of the 4-1BB agonist antibody H39E3-2.

[0336] Sequence ID 53 is the light chain variable region (VL) of the 4-1BB agonist antibody H39E3-2.

[0337] Sequence ID 54 is the amino acid sequence of human OX40.

[0338] Sequence ID 55 is the amino acid sequence of mouse OX40.

[0339] Sequence ID 56 is the heavy chain of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0340] Sequence ID 57 is the light chain of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0341] Sequence ID 58 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0342] Sequence ID 59 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0343] Sequence ID 60 is the heavy chain CDR1 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0344] Sequence ID 61 is the heavy chain CDR2 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0345] Sequence ID 62 is the heavy chain CDR3 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0346] Sequence ID 63 is the light chain CDR1 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0347] Sequence ID 64 is the light chain CDR2 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0348] Sequence ID 65 is the light chain CDR3 of the OX40 agonist monoclonal antibody tavorixizumab (MEDI-0562).

[0349] Sequence ID 66 is the heavy chain of the OX40 agonist monoclonal antibody 11D4.

[0350] Sequence ID 67 is the light chain of the OX40 agonist monoclonal antibody 11D4.

[0351] Sequence ID 68 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 11D4.

[0352] Sequence ID 69 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 11D4.

[0353] Sequence ID 70 is the heavy chain CDR1 of the OX40 agonist monoclonal antibody 11D4.

[0354] Sequence ID 71 is the heavy chain CDR2 of the OX40 agonist monoclonal antibody 11D4.

[0355] Sequence ID 72 is the heavy chain CDR3 of the OX40 agonist monoclonal antibody 11D4.

[0356] Sequence ID 73 is the light chain CDR1 of the OX40 agonist monoclonal antibody 11D4.

[0357] Sequence ID 74 is the light chain CDR2 of the OX40 agonist monoclonal antibody 11D4.

[0358] Sequence ID 75 is the light chain CDR3 of the OX40 agonist monoclonal antibody 11D4.

[0359] Sequence ID 76 is the heavy chain of the OX40 agonist monoclonal antibody 18D8.

[0360] Sequence ID 77 is the light chain of the OX40 agonist monoclonal antibody 18D8.

[0361] Sequence ID 78 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 18D8.

[0362] Sequence ID 79 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 18D8.

[0363] Sequence ID 80 is the heavy chain CDR1 of the OX40 agonist monoclonal antibody 18D8.

[0364] Sequence ID No. 81 is the heavy chain CDR2 of the OX40 agonist monoclonal antibody 18D8.

[0365] Sequence ID No. 82 is the heavy chain CDR3 of the OX40 agonist monoclonal antibody 18D8.

[0366] Sequence ID No. 83 is the light chain CDR1 of the OX40 agonist monoclonal antibody 18D8.

[0367] Sequence ID 84 is the light chain CDR2 of the OX40 agonist monoclonal antibody 18D8.

[0368] Sequence ID No. 85 is the light chain CDR3 of the OX40 agonist monoclonal antibody 18D8.

[0369] Sequence ID 86 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody Hu119-122.

[0370] Sequence ID 87 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody Hu119-122.

[0371] Sequence ID 88 is the heavy chain CDR1 of the OX40 agonist monoclonal antibody Hu119-122.

[0372] Sequence ID 89 is the heavy chain CDR2 of the OX40 agonist monoclonal antibody Hu119-122.

[0373] Sequence ID 90 is the heavy chain CDR3 of the OX40 agonist monoclonal antibody Hu119-122.

[0374] Sequence ID 91 is the light chain CDR1 of the OX40 agonist monoclonal antibody Hu119-122.

[0375] Sequence ID 92 is the light chain CDR2 of the OX40 agonist monoclonal antibody Hu119-122.

[0376] Sequence ID 93 is the light chain CDR3 of the OX40 agonist monoclonal antibody Hu119-122.

[0377] Sequence ID 94 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody Hu106-222.

[0378] Sequence ID 95 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody Hu106-222.

[0379] Sequence ID 96 is the heavy chain CDR1 of the OX40 agonist monoclonal antibody Hu106-222.

[0380] Sequence ID 97 is the heavy chain CDR2 of the OX40 agonist monoclonal antibody Hu106-222.

[0381] Sequence ID 98 is the heavy chain CDR3 of the OX40 agonist monoclonal antibody Hu106-222.

[0382] Sequence ID 99 is the light chain CDR1 of the OX40 agonist monoclonal antibody Hu106-222.

[0383] Sequence ID No. 100 is the light chain CDR2 of the OX40 agonist monoclonal antibody Hu106-222.

[0384] Sequence ID 101 is the light chain CDR3 of the OX40 agonist monoclonal antibody Hu106-222.

[0385] Sequence ID 102 is the amino acid sequence of the OX40 ligand (OX40L).

[0386] Sequence ID No. 103 is the soluble portion of the OX40L polypeptide.

[0387] Sequence ID No. 104 is an alternative soluble moiety of the OX40L polypeptide.

[0388] Sequence ID 105 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 008.

[0389] Sequence ID 106 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 008.

[0390] Sequence ID 107 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 011.

[0391] Sequence ID 108 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 011.

[0392] Sequence ID 109 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 021.

[0393] Sequence ID No. 110 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 021.

[0394] Sequence ID 111 is the heavy chain variable region (VH) of the OX40 agonist monoclonal antibody 023.

[0395] Sequence ID 112 is the light chain variable region (VL) of the OX40 agonist monoclonal antibody 023.

[0396] Sequence ID 113 is the heavy chain variable region (VH) of an OX40 agonist monoclonal antibody.

[0397] Sequence ID No. 114 is the light chain variable region (VL) of an OX40 agonist monoclonal antibody.

[0398] Sequence ID No. 115 is the heavy chain variable region (VH) of an OX40 agonist monoclonal antibody.

[0399] Sequence ID 116 is the light chain variable region (VL) of an OX40 agonist monoclonal antibody.

[0400] Sequence ID No. 117 is the heavy chain variable region (VH) of a humanized OX40 agonist monoclonal antibody.

[0401] Sequence ID 118 is the heavy chain variable region (VH) of a humanized OX40 agonist monoclonal antibody.

[0402] Sequence ID No. 119 is the light chain variable region (VL) of a humanized OX40 agonist monoclonal antibody.

[0403] Sequence ID No. 120 is the light chain variable region (VL) of a humanized OX40 agonist monoclonal antibody.

[0404] Sequence ID 121 is the heavy chain variable region (VH) of a humanized OX40 agonist monoclonal antibody.

[0405] Sequence ID No. 122 is the heavy chain variable region (VH) of a humanized OX40 agonist monoclonal antibody.

[0406] Sequence ID 123 is the light chain variable region (VL) of a humanized OX40 agonist monoclonal antibody.

[0407] Sequence ID No. 124 is the light chain variable region (VL) of a humanized OX40 agonist monoclonal antibody.

[0408] Sequence ID No. 125 is the heavy chain variable region (VH) of an OX40 agonist monoclonal antibody.

[0409] Sequence ID No. 126 is the light chain variable region (VL) of an OX40 agonist monoclonal antibody.

[0410] Sequence ID 127 is the amino acid sequence of human CD27.

[0411] Sequence ID 128 is the amino acid sequence of macaque CD27.

[0412] Sequence ID 129 is the heavy chain of the CD27 agonist monoclonal antibody varylumab (CDX-1127).

[0413] Sequence ID 130 is the light chain of the CD27 agonist monoclonal antibody varylumab (CDX-1127).

[0414] Sequence ID 131 is the heavy chain variable region (V) of the CD27 agonist monoclonal antibody varylumab (CDX-1127). H )

[0415] Sequence ID 132 is the light chain variable region (V) of the CD27 agonist monoclonal antibody varylumab (CDX-1127). L )

[0416] Sequence ID 133 is the heavy chain CDR1 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0417] Sequence ID 134 is the heavy chain CDR2 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0418] Sequence ID 135 is the heavy chain CDR3 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0419] Sequence ID 136 is the light chain CDR1 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0420] Sequence ID 137 is the light chain CDR2 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0421] Sequence ID 138 is the light chain CDR3 of the CD27 agonist monoclonal antibody varrilumab (CDX-1127).

[0422] Sequence ID 139 is the amino acid sequence of the CD27 ligand (CD70).

[0423] Sequence ID 140 is the soluble portion of the CD70 polypeptide.

[0424] Sequence ID 141 is an alternative soluble moiety of the CD70 polypeptide.

[0425] Sequence ID 142 is the amino acid sequence of human GITR (human tumor necrosis factor receptor superfamily member 18 (TNFRSF18) protein).

[0426] Sequence ID 143 is the amino acid sequence of mouse GITR (mouse tumor necrosis factor receptor superfamily member 18 (TNFRSF18) protein).

[0427] Sequence ID 144 is the amino acid sequence of HuN6C8 (glycosylated), a heavy chain variant of the 6C8 humanized GITR agonist monoclonal antibody having N (asparagine) in CDR2, which corresponds to Sequence ID 60 of U.S. Patent No. 7,812,135.

[0428] Sequence ID 145 is the amino acid sequence of HuN6C8 (non-glycosylated), a heavy chain variant of the 6C8 humanized GITR agonist monoclonal antibody having N (asparagine) in CDR2, which corresponds to Sequence ID 61 of U.S. Patent No. 7,812,135.

[0429] Sequence ID 146 is the amino acid sequence of HuQ6C8 (glycosylated), a heavy chain variant of the 6C8 humanized GITR agonist monoclonal antibody having Q (glutamine) in CDR2, which corresponds to Sequence ID 62 of U.S. Patent No. 7,812,135.

[0430] Sequence ID 147 is the amino acid sequence of HuQ6C8 (non-glycosylated), a heavy chain variant of the 6C8 humanized GITR agonist monoclonal antibody having Q (glutamine) in CDR2, which corresponds to Sequence ID 63 of U.S. Patent No. 7,812,135.

[0431] Sequence ID 148 is the amino acid sequence of the light chain of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 58 of U.S. Patent No. 7,812,135.

[0432] Sequence ID 149 is a leader sequence amino acid sequence that may be optionally included in GITR agonist monoclonal antibodies along with the amino acid sequences of Sequence ID 144, Sequence ID 145, Sequence ID 146, or Sequence ID 147.

[0433] Sequence ID 150 is a leader sequence amino acid sequence that may be optionally included in GITR agonist monoclonal antibodies along with the amino acid sequence of Sequence ID 148.

[0434] Sequence ID 151 is the amino acid sequence of the heavy chain variable region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 1 of U.S. Patent No. 7,812,135.

[0435] Sequence ID 152 is the amino acid sequence of the heavy chain variable region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 66 of U.S. Patent No. 7,812,135.

[0436] Sequence ID 153 is the amino acid sequence of the light chain variable region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 2 of U.S. Patent No. 7,812,135.

[0437] Sequence ID 154 is the amino acid sequence of the heavy chain CDR1 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 3 of U.S. Patent No. 7,812,135.

[0438] Sequence ID 155 is the amino acid sequence of the heavy chain CDR2 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 4 of U.S. Patent No. 7,812,135.

[0439] Sequence ID 156 is the amino acid sequence of the heavy chain CDR2 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 19 of U.S. Patent No. 7,812,135.

[0440] Sequence ID 157 is the amino acid sequence of the heavy chain CDR3 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 5 of U.S. Patent No. 7,812,135.

[0441] Sequence ID 158 is the amino acid sequence of the heavy chain CDR1 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 6 of U.S. Patent No. 7,812,135.

[0442] Sequence ID 159 is the amino acid sequence of the heavy chain CDR2 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 7 of U.S. Patent No. 7,812,135.

[0443] Sequence ID 160 is the amino acid sequence of the heavy chain CDR3 region of a 6C8 humanized GITR agonist monoclonal antibody, corresponding to Sequence ID 8 of U.S. Patent No. 7,812,135.

[0444] Sequence ID 161 is the amino acid sequence of HuN6C8 (glycosylated), a heavy chain variant of the 6C8 chimeric GITR agonist monoclonal antibody having N (asparagine) in CDR2, which corresponds to Sequence ID 23 of U.S. Patent No. 7,812,135.

[0445] Sequence ID 162 is the amino acid sequence of HuQ6C8 (non-glycosylated), a heavy chain variant of the 6C8 chimeric GITR agonist monoclonal antibody having Q (glutamine) in CDR2, which corresponds to Sequence ID 24 of U.S. Patent No. 7,812,135.

[0446] Sequence ID 163 is the amino acid sequence of the light chain of a 6C8 chimeric GITR agonist monoclonal antibody, corresponding to Sequence ID 22 of U.S. Patent No. 7,812,135.

[0447] Sequence ID 164 is the amino acid sequence of the 36E5 heavy chain variable region of the GITR agonist patent no. 8,709,424 in the United States.

[0448] Sequence ID No. 165 is the amino acid sequence of the GITR agonist 36E5 light chain variable region of U.S. Patent No. 8,709,424.

[0449] Sequence ID 166 is the amino acid sequence of the GITR agonist 3D6 heavy chain variable region of U.S. Patent No. 8,709,424.

[0450] Sequence ID No. 167 is the amino acid sequence of the GITR agonist 3D6 light chain variable region, U.S. Patent No. 8,709,424.

[0451] Sequence ID 168 is the amino acid sequence of the 61G6 heavy chain variable region of the GITR agonist patent no. 8,709,424 in the United States.

[0452] Sequence ID 169 is the amino acid sequence of the 61G6 light chain variable region of the GITR agonist, U.S. Patent No. 8,709,424.

[0453] Sequence ID No. 170 is the amino acid sequence of the 6H6 heavy chain variable region of the GITR agonist patent no. 8,709,424 in the United States.

[0454] Sequence ID No. 171 is the amino acid sequence of the 6H6 light chain variable region of the GITR agonist patent no. 8,709,424 in the United States.

[0455] Sequence ID 172 is the amino acid sequence of the heavy chain variable region of the GITR agonist 61F6, U.S. Patent No. 8,709,424.

[0456] Sequence ID 173 is the amino acid sequence of the light chain variable region of the GITR agonist 61F6, U.S. Patent No. 8,709,424.

[0457] Sequence ID 174 is the amino acid sequence of the heavy chain variable region of the GITR agonist 1D8, U.S. Patent No. 8,709,424.

[0458] Sequence ID No. 175 is the amino acid sequence of the light chain variable region of the GITR agonist 1D8, U.S. Patent No. 8,709,424.

[0459] Sequence ID 176 is the amino acid sequence of the heavy chain variable region of the GITR agonist 17F10, U.S. Patent No. 8,709,424.

[0460] Sequence ID 177 is the amino acid sequence of the light chain variable region of the GITR agonist 17F10, U.S. Patent No. 8,709,424.

[0461] Sequence ID 178 is the amino acid sequence of the heavy chain variable region of the GITR agonist 35D8, U.S. Patent No. 8,709,424.

[0462] Sequence ID 179 is the amino acid sequence of the light chain variable region of the GITR agonist 35D8, U.S. Patent No. 8,709,424.

[0463] Sequence ID 180 is the amino acid sequence of the heavy chain variable region of the GITR agonist 49A1, U.S. Patent No. 8,709,424.

[0464] Sequence ID 181 is the amino acid sequence of the light chain variable region of the GITR agonist 49A1, U.S. Patent No. 8,709,424.

[0465] Sequence ID 182 is the amino acid sequence of the GITR agonist 9E5 heavy chain variable region of U.S. Patent No. 8,709,424.

[0466] Sequence ID 183 is the amino acid sequence of the 9E5 light chain variable region of the GITR agonist, U.S. Patent No. 8,709,424.

[0467] Sequence ID 184 is the amino acid sequence of the GITR agonist 31H6 heavy chain variable region of U.S. Patent No. 8,709,424.

[0468] Sequence ID No. 185 is the amino acid sequence of the GITR agonist 31H6 light chain variable region of U.S. Patent No. 8,709,424.

[0469] Sequence ID No. 186 is the amino acid sequence of the 36E5 heavy chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0470] Sequence ID No. 187 is the amino acid sequence of the 36E5 light chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0471] Sequence ID No. 188 is the amino acid sequence of the 3D6 heavy chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0472] Sequence ID No. 189 is the amino acid sequence of the 3D6 light chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0473] Sequence ID No. 190 is the amino acid sequence of the 61G6 heavy chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0474] Sequence ID 191 is the amino acid sequence of the 61G6 light chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0475] Sequence ID 192 is the amino acid sequence of the 6H6 heavy chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0476] Sequence ID 193 is the amino acid sequence of the 6H6 light chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0477] Sequence ID 194 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 61F6, U.S. Patent No. 8,709,424.

[0478] Sequence ID 195 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 61F6, U.S. Patent No. 8,709,424.

[0479] Sequence ID 196 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 1D8, U.S. Patent No. 8,709,424.

[0480] Sequence ID 197 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 1D8, U.S. Patent No. 8,709,424.

[0481] Sequence ID 198 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 17F10, U.S. Patent No. 8,709,424.

[0482] Sequence ID 199 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 17F10, U.S. Patent No. 8,709,424.

[0483] Sequence ID No. 200 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 35D8, U.S. Patent No. 8,709,424.

[0484] Sequence ID No. 201 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 35D8, U.S. Patent No. 8,709,424.

[0485] Sequence ID No. 202 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 49A1, U.S. Patent No. 8,709,424.

[0486] Sequence ID 203 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 49A1, patent number 8,709,424 in the United States.

[0487] Sequence ID No. 204 is the amino acid sequence of the 9E5 heavy chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0488] Sequence ID No. 205 is the amino acid sequence of the light chain variable region of the humanized GITR agonist 9E5, U.S. Patent No. 8,709,424.

[0489] Sequence ID No. 206 is the amino acid sequence of the heavy chain variable region of the humanized GITR agonist 31H6, U.S. Patent No. 8,709,424.

[0490] Sequence ID No. 207 is the amino acid sequence of the 31H6 light chain variable region of the humanized GITR agonist, U.S. Patent No. 8,709,424.

[0491] Sequence ID No. 208 is the amino acid sequence of the GITR agonist 2155 variable heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0492] Sequence ID No. 209 is the amino acid sequence of the GITR agonist 2155 variable light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0493] Sequence ID No. 210 is the amino acid sequence of the GITR agonist 2155 humanized (HC1) heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0494] Sequence ID No. 211 is the amino acid sequence of the GITR agonist 2155 humanized (HC2) heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0495] Sequence ID 212 is the amino acid sequence of the GITR agonist 2155 humanized (HC3a) heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0496] Sequence ID No. 213 is the amino acid sequence of the humanized (HC3b) GITR agonist heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0497] Sequence ID No. 214 is the amino acid sequence of the humanized (HC4) GITR agonist heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0498] Sequence ID No. 215 is the amino acid sequence of the 2155 humanized (LC1) GITR agonist light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0499] Sequence ID 216 is the amino acid sequence of the 2155 humanized (LC2a) GITR agonist light chain in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0500] Sequence ID 217 is the amino acid sequence of the 2155 humanized (LC2b) GITR agonist light chain in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0501] Sequence ID No. 218 is the amino acid sequence of the 2155 humanized (LC3) GITR agonist light chain in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0502] Sequence ID No. 219 is the amino acid sequence of the GITR agonist 698 variable heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0503] Sequence ID No. 220 is the amino acid sequence of the GITR agonist 698 variable light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0504] Sequence ID No. 221 is the amino acid sequence of the GITR agonist 706 variable heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0505] Sequence ID No. 222 is the amino acid sequence of the GITR agonist 706 variable light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0506] Sequence ID 223 is the amino acid sequence of the GITR agonist 827 variable heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0507] Sequence ID No. 224 is the amino acid sequence of the GITR agonist 827 variable light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0508] Sequence ID No. 225 is the amino acid sequence of the GITR agonist 1718 variable heavy chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0509] Sequence ID No. 226 is the amino acid sequence of the GITR agonist 1718 variable light chain described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0510] Sequence ID No. 227 is the amino acid sequence of the GITR agonist 2155 heavy chain CDR3 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0511] Sequence ID No. 228 is the amino acid sequence of the GITR agonist 2155 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0512] Sequence ID No. 229 is the amino acid sequence of the GITR agonist 2155 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0513] Sequence ID No. 230 is the amino acid sequence of the GITR agonist 2155 light chain CDR3 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0514] Sequence ID No. 231 is the amino acid sequence of the GITR agonist 2155 light chain CDR2 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0515] Sequence ID 232 is the amino acid sequence of the GITR agonist 2155 light chain CDR1 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0516] Sequence ID No. 233 is the amino acid sequence of the GITR agonist 698 and 706 heavy chain CDR3 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0517] Sequence ID No. 234 is the amino acid sequence of the GITR agonist 698 and 706 heavy chain CDR2 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0518] Sequence ID No. 235 is the amino acid sequence of GITR agonist 698 and 706 heavy chain CDR1 from U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0519] Sequence ID No. 236 is the amino acid sequence of the GITR agonist 698 light chain CDR3 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0520] Sequence ID No. 237 is the amino acid sequence of GITR agonist 698, 706, 827 and 1649 light chain CDR2 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0521] Sequence ID No. 238 is the amino acid sequence of GITR agonist 698, 706, 827 and 1649 light chain CDR1 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0522] Sequence ID No. 239 is the amino acid sequence of GITR agonist 706, 827, and 1649 light chain CDR3 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0523] Sequence ID No. 240 is the amino acid sequence of GITR agonist 827 and heavy chain CDR3 1649 of U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0524] Sequence ID No. 241 is the amino acid sequence of the GITR agonist 827 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0525] Sequence ID No. 242 is the amino acid sequence of the GITR agonist 1649 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0526] Sequence ID No. 243 is the amino acid sequence of the GITR agonist 1718 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0527] Sequence ID No. 244 is the amino acid sequence of the GITR agonist 1718 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0528] Sequence ID No. 245 is the amino acid sequence of the GITR agonist 1718 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0529] Sequence ID No. 246 is the amino acid sequence of the GITR agonist 1718 light chain CDR3 in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0530] Sequence ID No. 247 is the amino acid sequence of the GITR agonist 1718 light chain CDR2, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0531] Sequence ID No. 248 is the amino acid sequence of the GITR agonist 1718 light chain CDR1, as described in U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0532] Sequence ID No. 249 is the amino acid sequence of GITR agonist 827 and heavy chain CDR1 1649 from U.S. Patent Application Publication No. 2013 / 0108641 A1.

[0533] Sequence ID No. 250 is the amino acid sequence of the GITR agonist 1D7 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0534] Sequence ID No. 251 is the amino acid sequence of the GITR agonist 1D7 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0535] Sequence ID 252 is the amino acid sequence of the GITR agonist 1D7 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0536] Sequence ID 253 is the amino acid sequence of the GITR agonist 1D7 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0537] Sequence ID 254 is the amino acid sequence of the GITR agonist 1D7 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0538] Sequence ID No. 255 is the amino acid sequence of the GITR agonist 1D7 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0539] Sequence ID No. 256 is the amino acid sequence of the GITR agonist 1D7 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0540] Sequence ID No. 257 is the amino acid sequence of the GITR agonist 1D7 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0541] Sequence ID 258 is the amino acid sequence of the GITR agonist 1D7 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0542] Sequence ID 259 is the amino acid sequence of the GITR agonist 1D7 light chain CDR3 in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0543] Sequence ID No. 260 is the amino acid sequence of the GITR agonist 33C9 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0544] Sequence ID No. 261 is the amino acid sequence of the GITR agonist 33C9 light chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0545] Sequence ID 262 is the amino acid sequence of the GITR agonist 33C9 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0546] Sequence ID No. 263 is the amino acid sequence of the GITR agonist 33C9 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0547] Sequence ID 264 is the amino acid sequence of the GITR agonist 33C9 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0548] Sequence ID No. 265 is the amino acid sequence of the GITR agonist 33C9 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0549] Sequence ID No. 266 is the amino acid sequence of the GITR agonist 33C9 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0550] Sequence ID No. 267 is the amino acid sequence of the GITR agonist 33C9 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0551] Sequence ID 268 is the amino acid sequence of the GITR agonist 33C9 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0552] Sequence ID No. 269 is the amino acid sequence of the GITR agonist 33C9 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0553] Sequence ID No. 270 is the amino acid sequence of the GITR agonist 33F6 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0554] Sequence ID No. 271 is the amino acid sequence of the GITR agonist 33F6 light chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0555] Sequence ID No. 272 ​​is the amino acid sequence of the GITR agonist 33F6 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0556] Sequence ID 273 is the amino acid sequence of the GITR agonist 33F6 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0557] Sequence ID No. 274 is the amino acid sequence of the GITR agonist 33F6 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0558] Sequence ID No. 275 is the amino acid sequence of the GITR agonist 33F6 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0559] Sequence ID No. 276 is the amino acid sequence of the GITR agonist 33F6 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0560] Sequence ID No. 277 is the amino acid sequence of the GITR agonist 33F6 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0561] Sequence ID No. 278 is the amino acid sequence of the GITR agonist 33F6 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0562] Sequence ID No. 279 is the amino acid sequence of the GITR agonist 33F6 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0563] Sequence ID No. 280 is the amino acid sequence of the GITR agonist 34G4 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0564] Sequence ID No. 281 is the amino acid sequence of the GITR agonist 34G4 light chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0565] Sequence ID No. 282 is the amino acid sequence of the GITR agonist 34G4 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0566] Sequence ID No. 283 is the amino acid sequence of the GITR agonist 34G4 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0567] Sequence ID No. 284 is the amino acid sequence of the GITR agonist 34G4 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0568] Sequence ID No. 285 is the amino acid sequence of the GITR agonist 34G4 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0569] Sequence ID No. 286 is the amino acid sequence of the GITR agonist 34G4 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0570] Sequence ID No. 287 is the amino acid sequence of the GITR agonist 34G4 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0571] Sequence ID No. 288 is the amino acid sequence of the GITR agonist 34G4 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0572] Sequence ID No. 289 is the amino acid sequence of the GITR agonist 34G4 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0573] Sequence ID No. 290 is the amino acid sequence of the GITR agonist 35B10 heavy chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0574] Sequence ID No. 291 is the amino acid sequence of the GITR agonist 35B10 light chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0575] Sequence ID 292 is the amino acid sequence of the GITR agonist 35B10 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0576] Sequence ID 293 is the amino acid sequence of the GITR agonist 35B10 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0577] Sequence ID No. 294 is the amino acid sequence of the GITR agonist 35B10 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0578] Sequence ID No. 295 is the amino acid sequence of the GITR agonist 35B10 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0579] Sequence ID No. 296 is the amino acid sequence of the GITR agonist 35B10 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0580] Sequence ID No. 297 is the amino acid sequence of the GITR agonist 35B10 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0581] Sequence ID No. 298 is the amino acid sequence of the GITR agonist 35B10 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0582] Sequence ID No. 299 is the amino acid sequence of the GITR agonist 35B10 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0583] Sequence ID No. 300 is the amino acid sequence of the GITR agonist 41E11 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0584] Sequence ID 301 is the amino acid sequence of the GITR agonist 41E11 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0585] Sequence ID 302 is the amino acid sequence of the GITR agonist 41E11 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0586] Sequence ID 303 is the amino acid sequence of the GITR agonist 41E11 variable light chain described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0587] Sequence ID 304 is the amino acid sequence of the GITR agonist 41E11 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0588] Sequence ID 305 is the amino acid sequence of the GITR agonist 41E11 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0589] Sequence ID 306 is the amino acid sequence of the GITR agonist 41E11 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0590] Sequence ID 307 is the amino acid sequence of the GITR agonist 41E11 light chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0591] Sequence ID 308 is the amino acid sequence of the GITR agonist 41E11 light chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0592] Sequence ID 309 is the amino acid sequence of the GITR agonist 41E11 light chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0593] Sequence ID No. 310 is the amino acid sequence of the GITR agonist 41G5 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0594] Sequence ID 311 is the amino acid sequence of the GITR agonist 41G5 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0595] Sequence ID 312 is the amino acid sequence of the GITR agonist 41G5 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0596] Sequence ID 313 is the amino acid sequence of the GITR agonist 41G5 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0597] Sequence ID 314 is the amino acid sequence of the GITR agonist 41G5 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0598] Sequence ID 315 is the amino acid sequence of the GITR agonist 41G5 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0599] Sequence ID 316 is the amino acid sequence of the GITR agonist 41G5 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0600] Sequence ID 317 is the amino acid sequence of the GITR agonist 41G5 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0601] Sequence ID 318 is the amino acid sequence of the GITR agonist 41G5 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0602] Sequence ID 319 is the amino acid sequence of the GITR agonist 41G5 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0603] Sequence ID No. 320 is the amino acid sequence of the GITR agonist 42A11 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0604] Sequence ID 321 is the amino acid sequence of the GITR agonist 42A11 light chain in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0605] Sequence ID 322 is the amino acid sequence of the GITR agonist 42A11 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0606] Sequence ID 323 is the amino acid sequence of the GITR agonist 42A11 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0607] Sequence ID 324 is the amino acid sequence of the GITR agonist 42A11 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0608] Sequence ID 325 is the amino acid sequence of the GITR agonist 42A11 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0609] Sequence ID 326 is the amino acid sequence of the GITR agonist 42A11 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0610] Sequence ID 327 is the amino acid sequence of the GITR agonist 42A11 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0611] Sequence ID 328 is the amino acid sequence of the GITR agonist 42A11 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0612] Sequence ID 329 is the amino acid sequence of the GITR agonist 42A11 light chain CDR3 in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0613] Sequence ID No. 330 is the amino acid sequence of the GITR agonist 44C1 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0614] Sequence ID 331 is the amino acid sequence of the GITR agonist 44C1 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0615] Sequence ID 332 is the amino acid sequence of the GITR agonist 44C1 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0616] Sequence ID 333 is the amino acid sequence of the GITR agonist 44C1 variable light chain described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0617] Sequence ID 334 is the amino acid sequence of the GITR agonist 44C1 heavy chain CDR1 described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0618] Sequence ID 335 is the amino acid sequence of the GITR agonist 44C1 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0619] Sequence ID 336 is the amino acid sequence of the GITR agonist 44C1 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0620] Sequence ID 337 is the amino acid sequence of the GITR agonist 44C1 light chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0621] Sequence ID 338 is the amino acid sequence of the GITR agonist 44C1 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0622] Sequence ID 339 is the amino acid sequence of the GITR agonist 44C1 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0623] Sequence ID No. 340 is the amino acid sequence of the GITR agonist 45A8 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0624] Sequence ID 341 is the amino acid sequence of the GITR agonist 45A8 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0625] Sequence ID 342 is the amino acid sequence of the GITR agonist 45A8 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0626] Sequence ID 343 is the amino acid sequence of the GITR agonist 45A8 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0627] Sequence ID 344 is the amino acid sequence of the GITR agonist 45A8 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0628] Sequence ID 345 is the amino acid sequence of the GITR agonist 45A8 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0629] Sequence ID 346 is the amino acid sequence of the GITR agonist 45A8 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0630] Sequence ID 347 is the amino acid sequence of the GITR agonist 45A8 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0631] Sequence ID 348 is the amino acid sequence of the GITR agonist 45A8 light chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0632] Sequence ID 349 is the amino acid sequence of the GITR agonist 45A8 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0633] Sequence ID 350 is the amino acid sequence of the GITR agonist 46E11 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0634] Sequence ID 351 is the amino acid sequence of the GITR agonist 46E11 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0635] Sequence ID 352 is the amino acid sequence of the GITR agonist 46E11 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0636] Sequence ID 353 is the amino acid sequence of the GITR agonist 46E11 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0637] Sequence ID 354 is the amino acid sequence of the GITR agonist 46E11 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0638] Sequence ID 355 is the amino acid sequence of the GITR agonist 46E11 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0639] Sequence ID 356 is the amino acid sequence of the GITR agonist 46E11 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0640] Sequence ID 357 is the amino acid sequence of the GITR agonist 46E11 light chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0641] Sequence ID 358 is the amino acid sequence of the GITR agonist 46E11 light chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0642] Sequence ID 359 is the amino acid sequence of the GITR agonist 46E11 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0643] Sequence ID 360 is the amino acid sequence of the GITR agonist 48H12 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0644] Sequence ID 361 is the amino acid sequence of the GITR agonist 48H12 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0645] Sequence ID 362 is the amino acid sequence of the GITR agonist 48H12 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0646] Sequence ID 363 is the amino acid sequence of the GITR agonist 48H12 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0647] Sequence ID 364 is the amino acid sequence of the GITR agonist 48H12 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0648] Sequence ID 365 is the amino acid sequence of the GITR agonist 48H12 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0649] Sequence ID 366 is the amino acid sequence of the GITR agonist 48H12 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0650] Sequence ID 367 is the amino acid sequence of the GITR agonist 48H12 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0651] Sequence ID 368 is the amino acid sequence of the GITR agonist 48H12 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0652] Sequence ID 369 is the amino acid sequence of the GITR agonist 48H12 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0653] Sequence ID No. 370 is the amino acid sequence of the GITR agonist 48H7 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0654] Sequence ID No. 371 is the amino acid sequence of the GITR agonist 48H7 light chain from U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0655] Sequence ID 372 is the amino acid sequence of the GITR agonist 48H7 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0656] Sequence ID 373 is the amino acid sequence of the GITR agonist 48H7 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0657] Sequence ID 374 is the amino acid sequence of the GITR agonist 48H7 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0658] Sequence ID 375 is the amino acid sequence of the GITR agonist 48H7 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0659] Sequence ID 376 is the amino acid sequence of the GITR agonist 48H7 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0660] Sequence ID 377 is the amino acid sequence of the GITR agonist 48H7 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0661] Sequence ID 378 is the amino acid sequence of the GITR agonist 48H7 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0662] Sequence ID 379 is the amino acid sequence of the GITR agonist 48H7 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0663] Sequence ID 380 is the amino acid sequence of the GITR agonist 49D9 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0664] Sequence ID 381 is the amino acid sequence of the GITR agonist 49D9 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0665] Sequence ID 382 is the amino acid sequence of the GITR agonist 49D9 variable heavy chain described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0666] Sequence ID 383 is the amino acid sequence of the GITR agonist 49D9 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0667] Sequence ID 384 is the amino acid sequence of the GITR agonist 49D9 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0668] Sequence ID 385 is the amino acid sequence of the GITR agonist 49D9 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0669] Sequence ID 386 is the amino acid sequence of the GITR agonist 49D9 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0670] Sequence ID 387 is the amino acid sequence of the GITR agonist 49D9 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0671] Sequence ID 388 is the amino acid sequence of the GITR agonist 49D9 light chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0672] Sequence ID 389 is the amino acid sequence of the GITR agonist 49D9 light chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0673] Sequence ID 390 is the amino acid sequence of the GITR agonist 49E2 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0674] Sequence ID 391 is the amino acid sequence of the GITR agonist 49E2 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0675] Sequence ID 392 is the amino acid sequence of the GITR agonist 49E2 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0676] Sequence ID 393 is the amino acid sequence of the GITR agonist 49E2 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0677] Sequence ID 394 is the amino acid sequence of the GITR agonist 49E2 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0678] Sequence ID 395 is the amino acid sequence of the GITR agonist 49E2 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0679] Sequence ID 396 is the amino acid sequence of the GITR agonist 49E2 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0680] Sequence ID 397 is the amino acid sequence of the GITR agonist 49E2 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0681] Sequence ID 398 is the amino acid sequence of the GITR agonist 49E2 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0682] Sequence ID 399 is the amino acid sequence of the GITR agonist 49E2 light chain CDR3 in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0683] Sequence ID No. 400 is the amino acid sequence of the GITR agonist 48A9 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0684] Sequence ID No. 401 is the amino acid sequence of the GITR agonist 48A9 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0685] Sequence ID 402 is the amino acid sequence of the GITR agonist 48A9 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0686] Sequence ID 403 is the amino acid sequence of the GITR agonist 48A9 variable light chain described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0687] Sequence ID 404 is the amino acid sequence of the GITR agonist 48A9 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0688] Sequence ID 405 is the amino acid sequence of the GITR agonist 48A9 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0689] Sequence ID No. 406 is the amino acid sequence of the GITR agonist 48A9 heavy chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0690] Sequence ID No. 407 is the amino acid sequence of the GITR agonist 48A9 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0691] Sequence ID No. 408 is the amino acid sequence of the GITR agonist 48A9 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0692] Sequence ID No. 409 is the amino acid sequence of the GITR agonist 48A9 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0693] Sequence ID No. 410 is the amino acid sequence of the GITR agonist 5H7 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0694] Sequence ID No. 411 is the amino acid sequence of the GITR agonist 5H7 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0695] Sequence ID 412 is the amino acid sequence of the GITR agonist 5H7 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0696] Sequence ID 413 is the amino acid sequence of the GITR agonist 5H7 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0697] Sequence ID 414 is the amino acid sequence of the GITR agonist 5H7 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0698] Sequence ID 415 is the amino acid sequence of the GITR agonist 5H7 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0699] Sequence ID No. 416 is the amino acid sequence of the GITR agonist 5H7 heavy chain CDR3 described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0700] Sequence ID No. 417 is the amino acid sequence of the GITR agonist 5H7 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0701] Sequence ID No. 418 is the amino acid sequence of the GITR agonist 5H7 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0702] Sequence ID 419 is the amino acid sequence of the GITR agonist 5H7 light chain CDR3 in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0703] Sequence ID No. 420 is the amino acid sequence of the GITR agonist 7A10 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0704] Sequence ID No. 421 is the amino acid sequence of the GITR agonist 7A10 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0705] Sequence ID 422 is the amino acid sequence of the GITR agonist 7A10 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0706] Sequence ID 423 is the amino acid sequence of the GITR agonist 7A10 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0707] Sequence ID 424 is the amino acid sequence of the GITR agonist 7A10 heavy chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0708] Sequence ID No. 425 is the amino acid sequence of the GITR agonist 7A10 heavy chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0709] Sequence ID 426 is the amino acid sequence of the GITR agonist 7A10 heavy chain CDR3, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0710] Sequence ID No. 427 is the amino acid sequence of the GITR agonist 7A10 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0711] Sequence ID 428 is the amino acid sequence of the GITR agonist 7A10 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0712] Sequence ID No. 429 is the amino acid sequence of the GITR agonist 7A10 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0713] Sequence ID No. 430 is the amino acid sequence of the GITR agonist 9H6 heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0714] Sequence ID No. 431 is the amino acid sequence of the GITR agonist 9H6 light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0715] Sequence ID 432 is the amino acid sequence of the GITR agonist 9H6 variable heavy chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0716] Sequence ID 433 is the amino acid sequence of the GITR agonist 9H6 variable light chain described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0717] Sequence ID 434 is the amino acid sequence of the GITR agonist 9H6 heavy chain CDR1, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0718] Sequence ID 435 is the amino acid sequence of the GITR agonist 9H6 heavy chain CDR2, as described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0719] Sequence ID 436 is the amino acid sequence of the GITR agonist 9H6 heavy chain CDR3 described in U.S. Patent Application Publication 2015 / 0064204 A1.

[0720] Sequence ID 437 is the amino acid sequence of the GITR agonist 9H6 light chain CDR1, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0721] Sequence ID 438 is the amino acid sequence of the GITR agonist 9H6 light chain CDR2, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0722] Sequence ID 439 is the amino acid sequence of the GITR agonist 9H6 light chain CDR3, as described in U.S. Patent Application Publication No. 2015 / 0064204 A1.

[0723] Sequence ID 440 is the amino acid sequence of the GITR ligand (GITRL).

[0724] Sequence ID 441 is the soluble portion of the GITRL polypeptide.

[0725] Sequence ID 442 is the amino acid sequence of human HVEM (CD270).

[0726] Sequence ID 443 is the amino acid sequence for the HVEM ligand (LIGHT).

[0727] Sequence ID 444 is the soluble portion of the LIGHT polypeptide.

[0728] Sequence ID No. 445 is an alternative soluble moiety of the LIGHT polypeptide.

[0729] Sequence ID 446 is an alternative soluble moiety of the LIGHT polypeptide.

[0730] Sequence ID 447 is the amino acid sequence of human CD95 isoform 1.

[0731] Sequence ID 448 is the amino acid sequence of human CD95 isoform 2.

[0732] Sequence ID 449 is the amino acid sequence of human CD95 isoform 3.

[0733] Sequence ID 450 is the amino acid sequence of human CD95 isoform 4.

[0734] Sequence ID 451 is the heavy chain variable region (V) of the CD95 agonist monoclonal antibody E09. H )

[0735] Sequence ID 452 is the light chain variable region (V) of the CD95 agonist monoclonal antibody E09. L )

[0736] Sequence ID 453 is the heavy chain CDR1 of the CD95 agonist monoclonal antibody E09.

[0737] Sequence ID 454 is the heavy chain CDR2 of the CD95 agonist monoclonal antibody E09.

[0738] Sequence ID 455 is the heavy chain CDR3 of the CD95 agonist monoclonal antibody E09.

[0739] Sequence ID 456 is the light chain CDR1 of the CD95 agonist monoclonal antibody E09.

[0740] Sequence ID 457 is the light chain CDR2 of the CD95 agonist monoclonal antibody E09.

[0741] Sequence ID 458 is the light chain CDR3 of the CD95 agonist monoclonal antibody E09.

[0742] Sequence ID 459 is the amino acid sequence of the CD95 ligand (CD95L).

[0743] Sequence ID 460 is the soluble portion of the CD95L polypeptide.

[0744] Sequence ID 461 is an alternative soluble moiety of the CD95L polypeptide.

[0745] Sequence ID 462 is an alternative soluble moiety of the CD95L polypeptide.

[0746] Sequence ID 463 is the heavy chain amino acid sequence of the PD-1 inhibitor nivolumab.

[0747] Sequence ID 464 is the light chain amino acid sequence of the PD-1 inhibitor nivolumab.

[0748] Sequence ID 465 is the heavy chain variable region (V) of the PD-1 inhibitor nivolumab. H ) This is the amino acid sequence.

[0749] Sequence ID 466 is the light chain variable region (V) of the PD-1 inhibitor nivolumab. L ) This is the amino acid sequence.

[0750] Sequence ID 467 is the heavy chain CDR1 amino acid sequence of the PD-1 inhibitor nivolumab.

[0751] Sequence ID 468 is the heavy chain CDR2 amino acid sequence of the PD-1 inhibitor nivolumab.

[0752] Sequence ID 469 is the heavy chain CDR3 amino acid sequence of the PD-1 inhibitor nivolumab.

[0753] Sequence ID 470 is the CDR1 amino acid sequence of the light chain of the PD-1 inhibitor nivolumab.

[0754] Sequence ID 471 is the CDR2 amino acid sequence of the light chain of the PD-1 inhibitor nivolumab.

[0755] Sequence ID 472 is the CDR3 amino acid sequence of the light chain of the PD-1 inhibitor nivolumab.

[0756] Sequence ID 473 is the heavy chain amino acid sequence of the PD-1 inhibitor pembrolizumab.

[0757] Sequence ID 474 is the light chain amino acid sequence of the PD-1 inhibitor pembrolizumab.

[0758] Sequence ID 475 is the heavy chain variable region (V) of the PD-1 inhibitor pembrolizumab. H ) This is the amino acid sequence.

[0759] Sequence ID 476 is the light chain variable region (V) of the PD-1 inhibitor pembrolizumab. L ) This is the amino acid sequence.

[0760] Sequence ID 477 is the heavy chain CDR1 amino acid sequence of the PD-1 inhibitor pembrolizumab.

[0761] Sequence ID 478 is the heavy chain CDR2 amino acid sequence of the PD-1 inhibitor pembrolizumab.

[0762] Sequence ID 479 is the heavy chain CDR3 amino acid sequence of the PD-1 inhibitor pembrolizumab.

[0763] Sequence ID 480 is the CDR1 amino acid sequence of the light chain of the PD-1 inhibitor pembrolizumab.

[0764] Sequence ID 481 is the CDR2 amino acid sequence of the light chain of the PD-1 inhibitor pembrolizumab.

[0765] Sequence ID 482 is the CDR3 amino acid sequence of the light chain of the PD-1 inhibitor pembrolizumab.

[0766] Sequence ID 483 is the heavy chain amino acid sequence of the PD-L1 inhibitor durvalumab.

[0767] Sequence ID 484 is the light chain amino acid sequence of the PD-L1 inhibitor durvalumab.

[0768] Sequence ID 485 is the heavy chain variable region (V) of the PD-L1 inhibitor durvalumab. H ) This is the amino acid sequence.

[0769] Sequence ID 486 is the light chain variable region (V) of the PD-L1 inhibitor durvalumab. L ) This is the amino acid sequence.

[0770] Sequence ID 487 is the heavy chain CDR1 amino acid sequence of the PD-L1 inhibitor durvalumab.

[0771] Sequence ID 488 is the heavy chain CDR2 amino acid sequence of the PD-L1 inhibitor durvalumab.

[0772] Sequence ID 489 is the heavy chain CDR3 amino acid sequence of the PD-L1 inhibitor durvalumab.

[0773] Sequence ID 490 is the CDR1 amino acid sequence of the light chain of the PD-L1 inhibitor durvalumab.

[0774] Sequence ID 491 is the CDR2 amino acid sequence of the light chain of the PD-L1 inhibitor durvalumab.

[0775] Sequence ID 492 is the CDR3 amino acid sequence of the light chain of the PD-L1 inhibitor durvalumab.

[0776] Sequence ID 493 is the heavy chain amino acid sequence of the PD-L1 inhibitor avelumab.

[0777] Sequence ID 494 is the light chain amino acid sequence of the PD-L1 inhibitor avelumab.

[0778] Sequence ID 495 is the heavy chain variable region (V) of the PD-L1 inhibitor avelumab. H ) This is the amino acid sequence.

[0779] Sequence ID 496 is the light chain variable region (V) of the PD-L1 inhibitor avelumab. L ) This is the amino acid sequence.

[0780] Sequence ID 497 is the heavy chain CDR1 amino acid sequence of the PD-L1 inhibitor avelumab.

[0781] Sequence ID 498 is the heavy chain CDR2 amino acid sequence of the PD-L1 inhibitor avelumab.

[0782] Sequence ID 499 is the heavy chain CDR3 amino acid sequence of the PD-L1 inhibitor avelumab.

[0783] Sequence ID 500 is the CDR1 amino acid sequence of the light chain of the PD-L1 inhibitor avelumab.

[0784] Sequence ID 501 is the CDR2 amino acid sequence of the light chain of the PD-L1 inhibitor avelumab.

[0785] Sequence ID 502 is the CDR3 amino acid sequence of the light chain of the PD-L1 inhibitor avelumab.

[0786] Sequence ID 503 is the heavy chain amino acid sequence of the PD-L1 inhibitor atezolizumab.

[0787] Sequence ID 504 is the light chain amino acid sequence of the PD-L1 inhibitor atezolizumab.

[0788] Sequence ID 505 is the heavy chain variable region (V) of the PD-L1 inhibitor atezolizumab. H ) This is the amino acid sequence.

[0789] Sequence ID 506 is the light chain variable region (V) of the PD-L1 inhibitor atezolizumab. L ) This is the amino acid sequence.

[0790] Sequence ID 507 is the heavy chain CDR1 amino acid sequence of the PD-L1 inhibitor atezolizumab.

[0791] Sequence ID 508 is the heavy chain CDR2 amino acid sequence of the PD-L1 inhibitor atezolizumab.

[0792] Sequence ID 509 is the heavy chain CDR3 amino acid sequence of the PD-L1 inhibitor atezolizumab.

[0793] Sequence ID 510 is the CDR1 amino acid sequence of the light chain of the PD-L1 inhibitor atezolizumab.

[0794] Sequence ID 511 is the CDR2 amino acid sequence of the light chain of the PD-L1 inhibitor atezolizumab.

[0795] Sequence ID 512 is the CDR3 amino acid sequence of the light chain of the PD-L1 inhibitor atezolizumab.

[0237] Detailed description of the invention

[0796] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. All patents and publications referenced herein are incorporated herein by reference in their entirety.

[0238] definition

[0797] As used herein, the terms “co-administration,” “co-administration,” “administered in combination with,” “administered in combination with,” “simultaneous,” and “concurrent” encompass the administration of two or more active pharmaceutical ingredients (in preferred embodiments of the present invention, for example, at least one TNFRSF agonist and multiple TILs) to a subject so that both active pharmaceutical ingredients and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different time points in separate compositions, or administration in a composition containing two or more active pharmaceutical ingredients. Simultaneous administration in separate compositions and administration in a composition containing both drugs are preferred.

[0239]

[0798] The term “rapid expansion culture” means an increase in the number of antigen-specific TILs of at least about 3 times (or 4 times, 5 times, 6 times, 7 times, 8 times, or 9 times) over a period of one week, more preferably at least about 10 times (or 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, or 90 times) over a period of one week, or most preferably at least about 100 times over a period of one week. Several rapid expansion culture protocols are described herein.

[0240]

[0799] In this specification, “tumor-infiltrating lymphocytes” or “TILs” means a population of cells that have migrated from the bloodstream into the tumor and were originally obtained as leukocytes. Examples of TILs include, but are not limited to, CD8+ cytotoxic T cells (lymphocytes), Th1 and Th17 CD4+ T cells, natural killer cells, dendritic cells, and M1 macrophages. TILs include both primary and secondary TILs. “Primary TILs” are obtained from patient tissue samples as outlined herein (sometimes referred to as “freshly recovered”), and “secondary TILs” are any TIL cell populations that have been cultured or grown as discussed herein, but are not limited to, bulk TILs and expanded culture TILs (“REP TILs” or “post-REP TILs”).

[0241]

[0800] In this specification, “cell population” (including TIL) means a large number of cells that share common traits. Generally, a population is roughly 1 × 10⁶ in number. 6 ~1 × 10 10 This is a range of individuals, and different TIL populations contain different numbers. For example, the initial growth of primary TILs in the presence of IL-2 is approximately 1 × 10⁻⁶. 8 This yields a bulk TIL population of individual cells. REP expansion cultures generally yield 1.5 × 10⁶ cells. 9 ~1.5×10 10 This is done so that individual cell populations for injection are provided.

[0242]

[0801] The term "central memory T cells" refers to a subset of T cells in humans that are CD45R0+ and constitutively express CCR7 (CCR7hi) and CD62L (CD62hi). The surface phenotype of central memory T cells also includes TCR, CD3, CD127 (IL-7R), and IL-15R. Transcription factors of central memory T cells include BCL-6, BCL-6B, MBD2, and BMI1. After TCR induction, central memory T cells primarily secrete IL-2 and CD40L as effector molecules. Central memory T cells are dominant in the CD4 compartment of the blood and are proportionally concentrated in lymph nodes and tonsils in humans.

[0243]

[0802] The term "anti-CD3 antibody" refers to an antibody or its variant, such as a monoclonal antibody, that is targeted against the CD3 receptor on the T cell antigen receptor of mature T cells and includes human, humanized, chimeric, or mouse antibodies. Anti-CD3 antibodies include OKT-3, also known as muromonab. Anti-CD3 antibodies also include the UHCT1 clone, also known as T3 and CD3ε. Other anti-CD3 antibodies include, for example, otelixizumab, teprizumab, and vizilizumab.

[0244]

[0803] The term "OKT-3" (also referred to herein as "OKT3") refers to monoclonal antibodies, biosimilars, or variants thereof, including human, humanized, chimeric, or mouse antibodies targeted against the CD3 receptor in the T cell antigen receptor of mature T cells, and includes commercially available forms such as OKT-3 (30 ng / mL, MACS GMP CD3 pure, Miltenyi Biotech, Inc., San Diego, CA, USA) and muromonab or their variants, conservative amino acid substitutions, glycoforms, or biosimilars. The amino acid sequences of the heavy and light chains of muromonab are shown in Table 1 (SEQ ID NOs: 1 and 2).

[0245] [Table 1]

[0246]

[0804] The term "IL-2" (also referred to herein as "IL2") refers to the T cell growth factor known as interleukin-2, and includes all forms of IL-2, including human and mammalian forms, conserved amino acid substitutions, glycoforms, biosimilars, and their variants. IL-2 is described, for example, in Nelson, J. Immunol. 2004, 172, 3983-88 and Malek, Annu. Rev. Immunol. 2008, 26, 453-79, the disclosures of which are incorporated herein by reference. The amino acid sequence of recombinant human IL-2 suitable for use in the present invention is shown in Table 2 (SEQ ID NO: 3). For example, the term IL-2 encompasses human recombinant forms of IL-2, including aldesleukin (PROLEUKIN, commercially available from multiple suppliers at 22 million IU per single-use vial) and recombinant IL-2 commercially supplied by CellGenix, Inc., Portsmouth, NH, USA (CELLGRO GMP) or ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ, USA (catalog number CYT-209-b), as well as other commercially available equivalents from other distributors. Aldesleukin (des-alanil-1, serine-125 human IL-2) is a non-glycosylated human recombinant IL-2 with a molecular weight of approximately 15 kDa. The amino acid sequence of aldesleukin suitable for use in the present invention is shown in Table 2 (SEQ ID NO: 4). The term IL-2 also encompasses pegylated forms of IL-2, including the pegylated IL-2 prodrug NKTR-214, available from Nektar Therapeutics, South San Francisco, CA, USA, as described herein. NKTR-214 and pegylated IL-2 suitable for use in the present invention are described in U.S. Patent Application Publication 2014 / 0328791 A1 and International Publication 2012 / 065086 A1 (these disclosures are incorporated herein by reference). Alternative forms of conjugate IL-2 suitable for use in the present invention are described in U.S. Patents 4,766,106, 5,206,344, 5,089,261 and 4902,502, the disclosures of which are incorporated herein by reference.Formulations of IL-2 suitable for use in the present invention are described in U.S. Patent No. 6,706,289, the disclosure of which is incorporated herein by reference.

[0247] [Table 2]

[0248]

[0805] The term "IL-4" (also referred to herein as "IL4") refers to the cytokine known as interleukin 4, which is produced by Th2 T cells and by eosinophils, basophils, and mast cells. IL-4 regulates the differentiation of naive helper T cells (Th0 cells) into Th2 T cells. Steinke and Borish, Respir. Res. 2001, 2, 66-70. Upon activation by IL-4, Th2 T cells subsequently produce further IL-4 in a positive feedback loop. IL-4 also stimulates B cell expansion culture and class II MHC expression, inducing class switching from B cells to IgE and IgG1 expression. Recombinant human IL-4 suitable for use in the present invention is commercially available from several suppliers, including ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ, USA (catalog number CYT-211) and ThermoFisher Scientific, Inc., Waltham, MA, USA (recombinant human IL-15 protein, catalog number Gibco CTP0043). The amino acid sequence of recombinant human IL-4 suitable for use in the present invention is shown in Table 2 (SEQ ID NO: 5).

[0249]

[0806] The term "IL-7" (also referred to herein as "IL7") refers to a glycosylated tissue-derived cytokine known as interleukin-7, which can be obtained from stromal and epithelial cells as well as dendritic cells. Fry and Mackall, Blood 2002, 99, 3892-904. IL-7 can stimulate T cell development. IL-7 binds to the IL-7 receptor, a heterodimer consisting of IIL-7 receptor alpha and a common gamma chain receptor, in a series of signals crucial for T cell development in the thymus and survival in the periphery. Recombinant human IL-7 suitable for use in this invention is commercially available from several suppliers, including ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ, USA (catalog no. CYT-254) and ThermoFisher Scientific, Inc., Waltham, MA, USA (human IL-7 recombinant protein, catalog no. Gibco PHC0071). Table 2 shows the amino acid sequence of recombinant human IL-7 suitable for use in the present invention (SEQ ID NO: 6).

[0250]

[0807] The term “IL-15” (also referred to herein as “IL15”) refers to the T cell growth factor known as interleukin-15 and includes all forms of IL-15, including human and mammalian forms, conserved amino acid substitutions, glycoforms, biosimilars and their variants. IL-15 is described, for example, in Fehniger and Caligiuri, Blood 2001, 97, 14-32, the disclosure of which is incorporated herein by reference. IL-15 shares β and γ signaling receptor subunits with IL-2. Recombinant human IL-15 is a single non-glycosylated polypeptide chain containing 114 amino acids (and N-terminal methionine) with a molecular weight of 12.8 kDa. Recombinant human IL-15 is commercially available from several suppliers, including ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ, USA (catalog number CYT-230-b) and ThermoFisher Scientific, Inc., Waltham, MA, USA (recombinant human IL-15 protein, catalog number 34-8159-82). The amino acid sequence of recombinant human IL-15 suitable for use in this invention is shown in Table 2 (SEQ ID NO: 7).

[0251]

[0808] The term “IL-21” (also referred to herein as “IL21”) refers to the pleiotropic cytokine protein known as interleukin-21 and includes all forms of IL-21, including human and mammalian forms, conserved amino acid substitutions, glycoforms, biosimilars and their variants. IL-21 is described, for example, in Spolski and Leonard, Nat. Rev. Drug. Disc. 2014, 13, 379-95, the disclosure of which is incorporated herein by reference. IL-21 is primarily produced by natural killer T cells and activated human CD4+ T cells. Recombinant human IL-21 is a single nonglycosylated polypeptide chain containing 132 amino acids with a molecular weight of 15.4 kDa. Recombinant human IL-21 is commercially available from several suppliers, including ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ, USA (catalog number CYT-408-b) and ThermoFisher Scientific, Inc., Waltham, MA, USA (recombinant human IL-21 protein, catalog number 14-8219-80). Table 2 shows the amino acid sequence of recombinant human IL-21 suitable for use in the present invention (SEQ ID NO: 8).

[0252]

[0809] Adenosine A2A receptor antagonists are called "A2aR antagonists" and "A 2A These receptors are referred to as "AdoR antagonists." They belong to the G protein-coupled receptor family and are distinct from the adenosine A1, adenosine A2B, and adenosine A3 receptor subfamilies.

[0253]

[0810] The term "CPI-444" refers to the compound 7-(5-methylfuran-2-yl)-3-[[6-[[(3S)-oxolan-3-yl]oxymethyl]pyridine-2-yl]methyl]triazolo[4,5-d]pyrimidine-5-amine, also known as siforadenanth. This compound is also known as "V81444". Its molecular formula is C 20 H 21It is N7O3. As used in this disclosure, the terms “CPI-444” or “siphoradenanth” encompass pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs of 7-(5-methylfuran-2-yl)-3-[[6-[[(3S)-oxolan-3-yl]oxymethyl]pyridine-2-yl]methyl]triazolo[4,5-d]pyrimidine-5-amine, respectively.

[0254]

[0811] The term "SCH58261" refers to the molecular formula C 18 H 15 The compound 2-(furan-2-yl)-7-phenethyl-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine-5-amine having N7O. As used in this disclosure, the term "SCH58261" encompasses pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs of 2-(furan-2-yl)-7-phenethyl-7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine-5-amine.

[0255]

[0812] The term "SYN115" refers to the molecular formula C 19 H 26 The compound 4-hydroxy-N-[4-methoxy-7-(4-morpholinyl)-2-benzothiazolyl]-4-methyl-1-piperidinecarboxamide has N4O4S. As used in this disclosure, the term "SYN115" encompasses pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs of 4-hydroxy-N-[4-methoxy-7-(4-morpholinyl)-2-benzothiazolyl]-4-methyl-1-piperidinecarboxamide.

[0256]

[0813] The term "ZM241385" refers to the molecular formula C 16 H 15The compound 4-(-2-[7-amino-2-{2-furyl}{1,2,4}triazolo{2,3-a}{1,3,5}triazine-5-yl-amino]ethyl)phenol having N7O2. As used in this disclosure, the term "ZM241385" encompasses pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs of 4-(-2-[7-amino-2-{2-furyl}{1,2,4}triazolo{2,3-a}{1,3,5}triazine-5-yl-amino]ethyl)phenol.

[0257]

[0814] The term "7MMB" refers to a family of compounds defined in the template, where X is C and R is selected from the group consisting of para-F, meta-F, para-CH3, 2,4-difluoro, 2,6-difluoro, 3,4-difluoro, 3,4-dimethoxy, meta-(2-methoxyethoxy), meta-(1,3-benzodioxole), para-Cl, para-CF3, para-CN and para-tert-butyl; where X is N and R is selected from the group consisting of para-F, meta-F, ortho-F, para-Cl, meta-CF3, 2,4-difluoro, 2,6-difluoro, 3,4-difluoro, meta-(2-methoxyethoxy), meta-(1,3-benzodioxole), para-CH3 and meta-OCH3. The term "7MMB" encompasses pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs of the genus disclosed in this template and in the following section on adenosine 2A receptor antagonists "7MMG".

[0258]

[0815] The term "in vivo" refers to events that occur within the body of mammals.

[0259]

[0816] The term "ex vivo" refers to events that occur outside the body of a mammal in an artificial environment.

[0260]

[0817] The term "in vitro" refers to events that occur in a test system. In vitro assays encompass cell-based assays in which live or dead cells may be used, and may also encompass cell-free assays in which intact cells are not used.

[0261]

[0818] The terms “effective dose” or “therapeutic effective dose” refer to the amount of a compound or combination of compounds described herein that is sufficient to achieve the intended use, including but not limited to disease treatment. The therapeutic effective dose may vary depending on the intended use (in vitro or in vivo), the subject and disease state being treated (e.g., the subject’s weight, age, and sex), the severity of the disease state, or the method of administration. This term also applies to doses that induce a specific response in target cells (e.g., reduced platelet adhesion and / or cell migration). The specific dose will vary depending on the specific compound selected, the administration regimen to be followed, whether the compound is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system through which the compound is carried.

[0262]

[0819] "Therapeutic effect," as used herein, encompasses therapeutic and / or preventive benefits. Preventive effect includes delaying or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0263]

[0820] The terms "QD," "qd," or "qd" mean once a day, once a day, or once every day. The terms "BID," "bid," or "bid" mean twice a day, twice a day, or twice every day. The terms "TID," "tid," or "tid" mean three times a day, three times a day, or three times every day. The terms "QID," "qid," or "qid" mean four times a day, four times a day, or four times every day. The term "QW" means once a week. The term "Q2W" means once every two weeks. The term "Q3W" means once every three weeks. The term "Q4W" means once every four weeks.

[0264]

[0821] The term "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be formed using inorganic and organic acids. Preferred inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Preferred organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases that can derive salts include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. The term "cocrystal" refers to molecular complexes derived from several cocrystal-forming agents known in the art. Unlike salts, cocrystals typically do not involve hydrogen transfer between the cocrystal and the drug, but instead involve intermolecular interactions such as hydrogen bonding, aromatic ring stacking, or dispersion forces between the cocrystal-forming organism and the drug in the crystalline structure.

[0265]

[0822] The terms “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” include all solvents, dispersants, coatings, antimicrobial and antifungal agents, isotonic and absorption retardants, and inactive components. The use of such pharmaceutically acceptable carriers or excipients for active pharmaceutical ingredients is well known in the art. Unless any conventional pharmaceutically acceptable carrier or excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the present invention is intended. Additional active pharmaceutical ingredients, such as other drugs, may also be incorporated into the compositions, processes, and methods described.

[0266]

[0823] The term “antigen” refers to a substance that induces an immune response. In some embodiments, an antigen is a molecule that can be conjugated by an antibody or T cell receptor (TCR) when presented by a major histocompatibility complex (MHC) molecule. As used herein, the term “antigen” also encompasses T cell epitopes. Antigens may be further recognized by the immune system. In some embodiments, an antigen can induce a humoral or cellular immune response that results in the activation of B lymphocytes and / or T lymphocytes. In some cases, this may require the antigen to contain or be conjugated to a Th cell epitope. An antigen may also have one or more epitopes (e.g., a B epitope and a T epitope). In some embodiments, an antigen preferably reacts with its corresponding antibody or TCR in a typically highly specific and selective manner, and does not react with a number of other antibodies or TCRs that may be induced by that antigen.

[0267]

[0824] The terms “antibody” and “antibody” refer to an entire immunoglobulin and any antigen-binding fragment (“antigen-binding moiety”) or their single chains. “Antibody” further refers to a glycoprotein or its antigen-binding moiety comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (V in this specification). HIt consists of a heavy chain constant region (abbreviated as CH1) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2 and CH3. Each light chain is a light chain variable region (V in this specification). L It consists of a (abbreviated as) and a light chain steady region. The light chain steady region is one domain, C L It is composed of the antibody V H and V L The region is called the Complementarity Determination Region (CDR) or Hypervariability Region (HVR), and can be further subdivided into hypervariability regions that can be dispersed into more conserved regions (called framework regions (FR)). H and V L It consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. The variable regions of the heavy and light chains contain binding domains that interact with the antigen epitope. The constant region of the antibody can mediate the binding of immunoglobulins to various cells of the immune system (e.g., effector cells) and to host tissues or factors including the first component (Clq) of the classical complement system.

[0268]

[0825] The terms “monoclonal antibody,” “mAb,” and “monoclonal antibody composition,” or their plural forms, refer to preparations of antibody molecules with a single molecular composition. Monoclonal antibody compositions exhibit a single binding specificity and affinity for a particular epitope. Monoclonal antibodies specific to the TNFRSF receptor can be produced using knowledge and techniques in the art, which involve injecting a suitable antigen into a test subject and then isolating hybridomas that express antibodies with the desired sequence or functional characteristics. The DNA encoding the monoclonal antibody can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of the monoclonal antibody). Hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA can be placed in an expression vector, which is then transfected into host cells such as Escherichia coli (E. coli) cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in recombinant host cells. Recombinant antibody production is described in more detail below.

[0269]

[0826] As used herein, the terms “antigen-binding moiety” or “antigen-binding fragment” (or simply “antibody moiety” or “fragment”) of an antibody refer to one or more fragments of an antibody that possess the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody may be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term “antigen-binding moiety” of an antibody include (i)V L , V H , C L (ii) a monovalent fragment consisting of the CH1 domain, the Fab fragment; (ii) a divalent fragment containing two Fab fragments linked by disulfide bridges in the hinge region, the F(ab')2 fragment; (iii) V H and Fd fragment consisting of CH1 domain; (iv) V of a single arm of antibody L and V H Fv fragment consisting of domains, (v)V H or V LIt may consist of domain-based antibody (dAb) fragments (Ward, et al., Nature, 1989, 341, 544-546); and (vi) isolated complementarity-determining regions (CDRs). Furthermore, it includes two domains of the Fv fragment, V L and V H These are encoded by separate genes, but they can be synthesized using recombination. L and V H Region pairs can be linked by synthetic linkers, which allow them to be constructed as a single protein chain forming a monovalent molecule known as a single-chain Fv (scFv); see, for example, Bird, et al., Science 1988, 242, 423-426; and Huston, et al., Proc.Natl.Acad.Sci.USA 1988, 85, 5879-5883). Such scFv antibodies are also intended to be encompassed within the terminology of the “antigen-binding portion” or “antigen-binding fragment” of an antibody. These antibody fragments are obtained using prior art known to those skilled in the art, and the fragments are screened for utility in the same manner as intact antibodies.

[0270]

[0827] As used herein, the term “human antibody” is intended to include antibodies having variable regions in which both the framework region and the CDR region are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region is also derived from a human germline immunoglobulin sequence. The human antibodies of the present invention may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or somatic mutation in vivo). As used herein, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as a mouse, has been transplanted into a human framework sequence.

[0271]

[0828] The term "human monoclonal antibody" refers to a single-binding specific antibody having variable regions in both its framework region and CDR region derived from human germline immunoglobulin sequences. In one embodiment, the human monoclonal antibody is obtained from a transgenic non-human animal, such as a transgenic mouse, and produced by a hybridoma containing B cells having a genome containing human heavy chain and light chain transgenes fused to immortalized cells.

[0272]

[0829] As used herein, the term “recombinant human antibody” includes all human antibodies prepared, expressed, produced or isolated by recombinant means, such as (a) antibodies isolated from animals (such as mice) that are transgenic or transchromosomes of human immunoglobulin genes or hybridomas prepared therefrom (as further described below), (b) antibodies isolated from host cells transformed to express human antibodies, e.g., transfectomas, (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, produced or isolated by any other means, including splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable regions in which the framework region and CDR region are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies can be subjected to in vitro mutagenesis (or in vivo somatic mutagenesis, if using animals transgenic for human Ig sequences), and therefore the V of recombinant antibodies H and V L The amino acid sequence of the region is from human germ cell line V H and V L While derived from and related to sequences, these sequences do not naturally exist within the in vivo human antibody germline repertoire.

[0273]

[0830] As used herein, “isotype” refers to the antibody class (e.g., IgM or IgG1) encoded by a heavy chain constant region gene.

[0274]

[0831] The phrases "antibody that recognizes an antigen" and "antibody that is specific to an antigen" are used interchangeably in this specification with the term "antibody that specifically binds to an antigen."

[0275]

[0832] The term "human antibody derivative" refers to any variant form of a human antibody, including a conjugate of an antibody with another active pharmaceutical ingredient or antibody. The terms "conjugate," "antibody-drug conjugate," "ADC," or "immunoconjugate" refer to an antibody or fragment thereof conjugated with another therapeutic portion, which can be conjugated to the antibodies described herein using methods available in the art.

[0276]

[0833] The terms "humanized antibody" and "humanized" refer to antibodies in which a CDR sequence derived from the germ cell line of another mammalian species, such as mouse, has been transplanted into a human framework sequence. Further modifications of the framework region may be made within the human framework sequence. Humanized non-human (e.g., mouse) antibodies are chimeric antibodies containing a minimal sequence derived from non-human immunoglobulin. For the most part, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's hypervariable region are replaced by residues from 15 hypervariable regions (donor antibodies) of a non-human species, such as mouse, rat, rabbit, or non-human primate, to have the desired specificity, affinity, and capability. In some cases, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may contain residues not found in either the recipient antibody or the donor antibody. These modifications are made to further improve antibody performance. Generally, humanized antibodies contain substantially all of at least one, typically two, variable domains, where all or substantially all of the hypervariable loops correspond to those of non-human immunoglobulins, and all or substantially all of the FR region is a human immunoglobulin sequence. Humanized antibodies also optionally contain at least a portion of the immunoglobulin constant region (Fc), typically that of human immunoglobulins. For further details, see Jones, et al., Nature 1986, 321, 522-525; Riechmann, et al., Nature 1988, 332, 323-329; and Presta, Curr.Op.Struct.Biol 1992, 2, 593-596. The TNFRSF agonists described herein may be modified to use any Fc variant known to confer improved (e.g., reduced) effector function and / or FcR binding.Fc variants include, for example, International Publication Nos. 1988 / 07089A1, 1996 / 14339A1, 1998 / 05787A1, 1998 / 23289A1, 1999 / 51642A1, 99 / 58572A1, 2000 / 09560A2, 2000 / 32767A1, 2000 / 42072A2, 2002 / 44215A2, and the same. No. 2002 / 060919A2, No. 2003 / 074569A2, No. 2004 / 016750A2, No. 2004 / 029207A2, No. 2004 / 035752A2, No. 2004 / 063351A2, 2004 / 074455A2, 2004 / 099249A2, 2005 / 040217A2, 2005 / 070963A1, 20 05 / 077981A2, 2005 / 092925A2, 2005 / 123780A2, 2006 / 019447A1, 2006 / 047350A2 and 2006 / 085967A2; and US Patent Nos. 5,648,260, 5,739,277, 5,834,250, 5,869,046, 6,096,871, 6, This may include any one of the amino acid substitutions disclosed in Patent Nos. 121,022, 6,194,551, 6,242,195, 6,277,375, 6,528,624, 6,538,124, 6,737,056, 6,821,505, 6,998,253; and 7,083,784, the disclosures of which are incorporated herein by reference.

[0277]

[0834] The term "chimeric antibody" is intended to refer to an antibody in which the variable region sequence originates from one species and the constant region sequence originates from another species, such as an antibody in which the variable region sequence originates from a mouse antibody and the constant region sequence originates from a human antibody.

[0278]

[0835] A "diabody" is a small antibody fragment that has two antigen-binding sites. The fragment is the same polypeptide chain (V H -V L or V L -V H) Light chain variable domain (V L ) connected to the heavy chain variable domain (V H ) includes. By using a linker that is too short to pair two domains on the same chain, the domain is forced to pair with a complementary domain on another chain, generating two antigen-binding sites. Diabodies are described more extensively, for example, in European Patent No. 404,097, International Publication No. 93 / 11161; and Bolliger, et al., Proc. Natl. Acad. Sci. USA 1993, 90, 6444-6448.

[0279]

[0836] The term "glycosylation" refers to modified derivatives of antibodies. Agricosylated antibodies lack glycosylation. Glycosylation can be modified, for example, to increase the affinity of an antibody to an antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites in the antibody sequence. For example, one or more amino acid substitutions can be made, resulting in the removal of one or more variable region framework glycosylation sites, thereby eliminating glycosylation at those sites. As described in U.S. Patents 5,714,350 and 6,350,861, aglycosylation can increase the affinity of an antibody to an antigen. Additionally or alternatively, antibodies with altered types of glycosylation can be produced, such as low-fucosylated antibodies with a reduced amount of fucosyl residues or antibodies with increased bisecting GlcNac structures. Such modified glycosylation patterns have been demonstrated to increase the capabilities of antibodies. Such carbohydrate modifications can be achieved, for example, by altering the glycosylation mechanism to express antibodies in host cells. Cells with modified glycosylation mechanisms have been described in the Art and can be used as host cells to express the recombinant antibodies of the present invention and thereby produce antibodies having the modified glycosylation. For example, the cell lines Ms704, Ms705, and Ms709 lack the fucosyltransferase gene, FUT8 (alpha(1,6)fucosyltransferase), so antibodies expressed in the Ms704, Ms705, and Ms709 cell lines lack fucose on their carbohydrates. The Ms704, Ms705, and Ms709 FUT8- / - cell lines were generated by targeting and disrupting the FUT8 gene in CHO / DG44 cells using two substitution vectors (see, e.g., U.S. Patent Application Publication No. 2004 / 0110704 or Yamane-Ohnuki, et al, Biotechnol.Bioeng., 2004, 87, 614-622).As another example, European Patent No. 1,176,195 describes a cell line having a functionally disrupted FUT8 gene encoding a fucosyltransferase such that antibodies expressed in such cell lines exhibit low fucosylation by reducing or eliminating an alpha-1,6 linkage-related enzyme, and also describes cell lines with low or no enzymatic activity in adding fucose to N-acetylglucosamine bound to the Fc region of antibodies, such as the rat myeloma cell line YB2 / 0 (ATCC CRL 1662). International Publication No. 03 / 035835 describes the mutant CHO cell line, Lec13 cells, which have a reduced ability to bind fucose to Asn(297) linked carbohydrates and result in low fucosylation of antibodies expressed in its host cells (see also Shields, et al., J. Biol Chem. 2002, 277, 26733-26740). International Publication No. 99 / 54342 describes a cell line engineered to express a glycoprotein-modified glycosyltransferase (e.g., beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such that the antibody expressed in the engineered cell line exhibits an increased bisecting GlcNac structure, resulting in increased ADCC activity of the antibody (see also Umana, et al., Nat. Biotech. 1999, 17, 176-180). Alternatively, the fucose residues of the antibody can be cleaved using a fucosidase enzyme. For example, alpha-L-fucosidase, a fucosidase, removes fucosyl residues from the antibody, as described in Tarentino, et al., Biochem. 1975, 14, 5516-5523.

[0280]

[0837] "Pegylation" refers to a modified antibody or fusion protein or a fragment thereof that reacts with PEG, such as a reactive ester or aldehyde derivative of polyethylene glycol (PEG), under conditions such that one or more PEG groups become attached to the antibody or antibody fragment. Pegylation can, for example, increase the biological (e.g., serum) half-life of an antibody. Preferably, pegylation is effected via an acylation or alkylation reaction with a reactive PEG molecule (or similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any form of PEG that has been used to derivatize other proteins, such as mono(C1-C 10 )alkoxy- or aryloxy-...

Claims

1. A method for expanding the culture of tumor-infiltrating lymphocytes (TILs) into a therapeutic TIL population, (a) A step of processing a tumor sample containing a first TIL population into multiple tumor fragments; (b) Adding the tumor fragment to the closed system; (c) A first expansion culture is performed to produce a second TIL population by culturing the first TIL population in a cell culture medium containing IL-2 and optionally OKT-3, wherein the first expansion culture is performed in a closed container providing a first gas-permeable surface area, the first expansion culture is performed for about 3 to 14 days to obtain the second TIL population, and the transition from step (b) to step (c) occurs without opening the system; (d) A second expansion culture is performed to produce a third TIL population by supplementing the cell culture medium of the second TIL population with additional IL-2, OKT-3, and antigen-presenting cells (APCs), wherein the second expansion culture is performed for approximately 7 to 14 days to obtain the third TIL population, the third TIL population being a therapeutic TIL population, the second expansion culture is performed in a closed container providing a second gas-permeable surface area, and the transition from step (c) to step (d) occurs without opening the system; (e) A step of collecting the therapeutic TIL population obtained from step (d), wherein the transition from step (d) to step (e) occurs without opening the system; and (f) A step of transferring the recovered TIL mass from step (e) to an infusion bag, wherein the transition from step (e) to (f) occurs without opening the system. Includes, The cell culture medium of step (c) and / or step (d) contains an adenosine 2A receptor (A2aR) antagonist containing CPI-444. method.

2. The method according to claim 1, wherein the adenosine 2A receptor (A2aR) antagonist further comprises vipardenant, SCH58261, ZM241385, SCH420814, SYN115, 8-CSC, KW-6002, A2A receptor antagonist 1, ADZ4635, ST4206, KF21213, SCH412348, 7MMG-49 or pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs thereof and combinations thereof.

3. The adenosine 2A receptor (A2aR) antagonist is added to the cell culture medium in step (c) at intervals selected from the group consisting of daily, every two days, every three days, every four days, every five days, every six days, every seven days and every two weeks during the first expansion culture, and / or the A2aR antagonist is added to the cell culture medium in step (d) at intervals selected from the group consisting of daily, every two days, every three days, every four days, every five days, every six days, every seven days and every two weeks during the second expansion culture. The method according to claim 1 or 2, wherein the adenosine 2A receptor (A2aR) antagonist is optionally added in the cell culture medium of step (c) or in a concentration sufficient to achieve a concentration of 0.01 μg / mL to 500 μg / mLM in the cell culture medium of step (c), and optionally added in the cell culture medium of step (c) or in a concentration sufficient to achieve a concentration of 1 μg / mL to 100 μg / mL.

4. The method according to any one of claims 1 to 3, wherein IL-2 is present in the cell culture medium of step (c) at an initial concentration of 10 to 6000 IU / mL, optionally, IL-2 is present in the cell culture medium of step (c) at an initial concentration of 3000 IU / mL, or IL-2 is present in the cell culture medium of step (c) at an initial concentration of 800 to 1100 IU / mL, optionally, IL-2 is present in the cell culture medium of step (c) at an initial concentration of 1000 IU / mL.

5. The method according to any one of claims 1 to 4, wherein IL-2 is present in the cell culture medium of step (d) at an initial concentration of 10 to 6000 IU / mL, optionally, IL-2 is present in the cell culture medium of step (d) at an initial concentration of 3000 IU / mL, or IL-2 is present in the cell culture medium of step (d) at an initial concentration of 800 to 1100 IU / mL, optionally, IL-2 is present in the cell culture medium of step (d) at an initial concentration of 1000 IU / mL.

6. The method according to any one of claims 1 to 5, wherein IL-15 is present in the cell culture medium of step (c), and / or IL-15 is present in the cell culture medium of step (d), and optionally IL-15 is present in the cell culture medium of step (c) at an initial concentration of 5 ng / mL to 20 ng / mL, and / or IL-15 is present in the cell culture medium of step (d) at an initial concentration of 5 ng / mL to 20 ng / mL.

7. The method according to any one of claims 1 to 6, wherein IL-21 is present in the cell culture medium of step (c), and / or IL-21 is present in the cell culture medium of step (d), and optionally IL-21 is present in the cell culture medium of step (c) at an initial concentration of 5 ng / mL to 20 ng / mL, and / or IL-21 is present in the cell culture medium of step (d) at an initial concentration of 5 ng / mL to 20 ng / mL.

8. The method according to any one of claims 1 to 7, wherein the OKT-3 antibody is present in the cell culture medium of step (d) at an initial concentration of 10 ng / mL to 60 ng / mL, and optionally, the OKT-3 antibody is present in the cell culture medium of step (d) at an initial concentration of 30 ng / mL.

9. The method according to any one of claims 1 to 8, wherein the first expansion culture is carried out using a gas-permeable container, and / or the second expansion culture is carried out using a gas-permeable container.

10. The method according to any one of claims 1 to 9, wherein the tumor sample is from any solid tumor, including a primary tumor, an invasive tumor, or a metastatic tumor, and optionally, the tumor sample is from any solid tumor.

11. The method according to any one of claims 1 to 10, wherein the tumor is excised from a patient who was treated with a PD-1 inhibitor or a PD-L1 inhibitor before the tumor was excised.

12. The method according to claim 11, wherein the PD-1 inhibitor or PD-L1 inhibitor is selected from the group consisting of nivolumab, pembrolizumab, durvalumab, atezolizumab, avelumab, and their fragments, derivatives, variants, biosimilars, and combinations.

13. The method according to any one of claims 1 to 10, wherein the cell culture medium of step (c) further comprises IL-4, IL-7, or a combination thereof, and / or the cell culture medium of step (d) further comprises IL-4, IL-7, or a combination thereof.

14. The method according to any one of claims 1 to 11, wherein the first expansion culture is carried out over a period of 11 days or less, and / or the second expansion culture is carried out over a period of 11 days or less.

15. The third TIL group is CD4 in the second TIL group. ＋ CD8 for TIL ＋ Compared to the reference ratio of TIL, CD4 ＋ CD8 for TIL ＋ The method according to any one of claims 1 to 12, which shows the increased ratio of TIL.

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