Combination therapy with BCMA-directed T cell therapy

Abstract

Provided herein are methods, compositions and uses for treating subjects with diseases and conditions, such as those involving or associated with B-cell maturation antigen (BCMA).In some aspects, the disclosure relates to certain combination therapies, including for treating subjects with multiple myeloma, that include administering cell therapy, e.g., BCMA-targeted T cell therapy, in combination with immunomodulatory agents and other agents, e.g., dexamethasone.In some embodiments, the multiple myeloma is relapsed or refractory multiple myeloma.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 176,196, filed April 16, 2021, the entire contents of which are incorporated herein by reference for all purposes. Incorporation by reference to sequence listing

[0002] This application is being filed with an electronic Sequence Listing, which is provided as a file entitled 735042025340SeqList.txt, created on April 10, 2022, and which is 328 kilobytes in size. The information in the electronic Sequence Listing is incorporated by reference in its entirety. Field

[0003] In some aspects, the present disclosure relates to methods, compositions, and uses for treating subjects with diseases and conditions, such as those involving or associated with B-cell maturation antigen (BCMA). In some aspects, the present disclosure relates to certain combination therapies comprising administration of cell therapy, e.g., BCMA-targeted T-cell therapy, in combination with immunomodulatory agents and other agents, e.g., dexamethasone, including for the treatment of subjects with multiple myeloma. In some embodiments, the multiple myeloma is relapsed or refractory multiple myeloma. [Background technology]

[0004] For example, various strategies for immunotherapy are available, such as administering engineered T cells for adoptive therapy.For example, strategies are available for engineering T cells that express genetically engineered antigen receptors, such as chimeric antigen receptors (CARs), and administering compositions containing such cells to subjects.Improved strategies are needed to improve the efficacy of cells, for example, to improve the persistence, activity and / or proliferation of cells when administered to subjects.Methods, compositions and kits that meet such needs are provided. Summary of the Invention

[0005] In some embodiments, provided herein are methods of treating a BCMA-expressing cancer, the method comprising: (a) administering to a subject having a BCMA-expressing cancer a T cell therapy comprising a dose of engineered T cells expressing a cancer-directed chimeric antigen receptor (CAR); (b) administering to the subject an immunomodulatory compound that is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; and (c) administering dexamethasone to the subject.

[0006] In some of the provided methods, the cancer is multiple myeloma. In some of the provided methods, the cancer is relapsed or refractory multiple myeloma (R / R MM).

[0007] In some of any of the provided embodiments, the CAR specifically binds to BCMA.

[0008] In some embodiments, a method of treating multiple myeloma comprises: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM) a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA; and (b) administering to the subject a T cell therapy comprising the following structure: [ka] or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; and (c) administering dexamethasone to a subject.

[0009] In some of the provided methods, the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione or a pharmaceutically acceptable salt thereof.

[0010] In some of the provided methods, the immunomodulatory compound and dexamethasone are administered simultaneously. In some of the provided methods, administration of the immunomodulatory compound and administration of dexamethasone are initiated simultaneously. In some of the provided methods, administration of the immunomodulatory compound and administration of dexamethasone are initiated on the same day. In some of the provided methods, doses of the immunomodulatory compound and dexamethasone are administered on the same day.

[0011] In some of the provided methods, administration of the immunomodulatory compound is initiated prior to administration of the T cell therapy. In some of the provided methods, administration of the immunomodulatory compound is initiated 0-30 days, 0-15 days, 0-6 days, 0-96 hours, 0-24 hours, 0-12 hours, 0-6 hours, 0-2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours after administration of the T cell therapy. Between 6 hours and 24 hours, 6 hours and 12 hours, 12 hours and 30 days, 12 hours and 15 days, 12 hours and 6 days, 12 hours and 96 hours, 12 hours and 24 hours, 24 hours and 30 days, 24 hours and 15 days, 24 hours and 6 days, 24 hours and 96 hours, 96 hours and 30 days, 96 hours and 15 days, 96 hours and 6 days, 6 days and 30 days, 6 days and 15 days, or 15 days and 30 days before or approximately thereafter.

[0012] In some of any of the provided methods, the immunomodulatory compound is administered concurrently with the administration of the T cell therapy.

[0013] In some of the provided methods, administration of the immunomodulatory compound is initiated following administration of the T cell therapy. In some of the provided methods, administration of the immunomodulatory compound is initiated up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after administration of the T cell therapy.

[0014] In some of any of the provided methods, administration of the immunomodulatory compound is initiated at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or about at least 12 days, at least or about at least 14 days, at least or about at least 15 days, at least or about at least 21 days, at least or about at least 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days, or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after administration of the T cell therapy.

[0015] In some of the provided methods, the immunomodulatory compound is or comprises a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione. In some of the provided methods, the immunomodulatory compound is or comprises a hydrate of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione. In some of the provided methods, the immunomodulatory compound is or comprises a solvate of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione. In some of the provided methods, the immunomodulatory compound is or includes (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione.

[0016] In some of the provided methods, the dexamethasone is administered in a low dose.

[0017] In some of any of the provided methods, prior to the method, the subject has not previously been administered an immunomodulatory compound.

[0018] Also provided herein, in some embodiments, are methods of treating a BCMA-expressing cancer, the method comprising: (a) administering to a subject having a BCMA-expressing cancer a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) directed against the cancer; and (b) administering to the subject a standard triplet regimen.

[0019] In some of the provided methods, the cancer is multiple myeloma. In some of the provided methods, the cancer is relapsed or refractory multiple myeloma (R / R MM).

[0020] In some of any of the provided embodiments, the CAR specifically binds to BCMA.

[0021] Also provided herein, in some embodiments, are methods of treating multiple myeloma, comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM) a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds BCMA; and (b) administering to the subject a standard triplet regimen.

[0022] In some of any of the provided methods, administration of the standard triplet regimen is initiated subsequent to administration of the T cell therapy.

[0023] Also provided herein, in some embodiments, are methods of treating a BCMA-expressing cancer, comprising administering a standard triplet regimen to a subject having a BCMA-expressing cancer, wherein at the time administration of the standard triplet regimen is initiated, the subject has previously received T cell therapy, and the T cell therapy comprises a dose of engineered T cells expressing a chimeric antigen receptor (CAR) directed against the cancer.

[0024] In some of the provided methods, the cancer is multiple myeloma. In some of the provided methods, the cancer is relapsed or refractory multiple myeloma (R / R MM).

[0025] In some of any of the provided embodiments, the CAR specifically binds to BCMA.

[0026] Also provided herein, in some embodiments, are methods of treating multiple myeloma, comprising administering a standard triplet regimen to a subject with relapsed or refractory multiple myeloma (R / R MM), wherein at the time administration of the standard triplet regimen is initiated, the subject has previously received T cell therapy, and the T cell therapy comprises a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds BCMA.

[0027] In some of the provided methods, the standard triplet regimen includes administration of daratumumab, pomalidomide and dexamethasone.

[0028] In some of any of the provided methods, prior to the method, the subject has not previously been administered daratumumab in combination with pomalidomide.

[0029] In some of the provided methods, the standard triplet regimen includes administration of pomalidomide, bortezomib, and dexamethasone.

[0030] In some of any of the provided methods, prior to the method, the subject has not previously been administered pomalidomide in combination with bortezomib.

[0031] In some of the provided methods, the dexamethasone is administered in a low dose.

[0032] In some of the provided methods, administration of the standard triplet regimen is initiated prior to administration of the T cell therapy. In some of the provided methods, administration of the standard triplet regimen is initiated 0-30 days, 0-15 days, 0-6 days, 0-96 hours, 0-24 hours, 0-12 hours, 0-6 hours, or 0-2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 6 hours, or 6 hours after administration of the T cell therapy. In some of the provided methods, administration of the standard triplet regimen is initiated about 96 hours, 6 hours to 24 hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days to 30 days prior to administration of the T cell therapy. In some of the provided methods, administration of the standard triplet regimen is initiated about 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour or less prior to administration of the T cell therapy.

[0033] In some of any of the provided methods, the standard triplet regimen is administered concurrently with the administration of the T cell therapy.

[0034] In some of the provided methods, administration of the standard triplet regimen is initiated following administration of the T cell therapy. In some of the provided methods, administration of the standard triplet regimen is initiated up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after administration of the T cell therapy. In some of the provided methods, administration of the standard triplet regimen begins at or about 56 days (8 weeks) after administration of the T cell therapy. In some of the provided methods, T cell therapy is administered on day 1 of the method, and administration of the standard triplet regimen begins on or about month 3, day 1 (M3D1) of the method.

[0035] In some of any of the provided methods, administration of the standard triplet regimen is initiated at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or about at least 12 days, at least or about at least 14 days, at least or about at least 15 days, at least or about at least 21 days, at least or about at least 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days, or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after administration of the T cell therapy.

[0036] In some embodiments, administration of the standard triplet regimen occurs between 1 day and 360 days, between 1 day and 240 days, between 1 day and 180 days, between 1 day and 120 days, between 1 day and 90 days, between 1 day and 60 days, between 2 days and 360 days, between 2 days and 240 days, between 2 days and 180 days, between 2 days and 120 days, between 2 days and 90 days, between 2 days and 60 days, between 3 days and 360 days, between 3 days and 240 days, between 3 days and 180 days, between 3 days and 120 days, between 3 days and 90 days, between 3 days and 60 days, between 4 days and 5 days after administration of the T cell therapy. 360 days later, 4 days later - 240 days later, 4 days later - 180 days later, 4 days later - 120 days later, 4 days later - 90 days later, 4 days later - 60 days later, 5 days later - 360 days later, 5 days later - 240 days later, 5 days later - 180 days later, 5 days later - 120 days later, 5 days later - 90 days later, 5 days later 60 days later, 6 days later - 360 days later, 6 days later - 240 days later, 6 days later - 180 days later, 6 days later - 120 days later, 6 days later - 90 days later, 6 days later - 60 days later, 7 days later - 360 days later, 7 days later - 240 days later, 7 days later - 180 days later, 7 days later - 120 days later, 7 days later - 9 0 days later, 7 days later - 60 days later, 14 days later - 360 days later, 14 days later - 240 days later, 14 days later - 180 days later, 14 days later - 120 days later, 14 days later - 90 days later, 14 days later - 60 days later, 21 days later - 360 days later, 21 days later - 240 days later, 21 days later - 180 Days later, 21 days to 120 days, 21 days to 90 days, 21 days to 60 days, 28 days to 360 days, 28 to 240 days, 28 to 180 days, 28 to 120 days, 28 to 90 days, 28 to 60 days, 35 to 360 days , 35 to 240 days, 35 to 180 days, 35 to 120 days, 35 to 90 days, 35 to 60 days, 42 to 360 days, 42 to 240 days, 42 to 180 days, 42 to 120 days, 42 to 90 days, 42 to 60 days, 49 to 360 days, 49 to 240 days, 49 to 180 days, 49 to 120 days, 49 to 90 days, or 49 to 60 days, or any approximate interval therebetween (inclusive of both ends).

[0037] In some of the provided methods, the dose of genetically engineered T cells is 5 x 10 7 or approximate number of CAR+ T cells ~1 x 10 9In some of the provided methods, the dose of engineered T cells is between 1 x 10 and 1 x 10 8 or approximate number of CAR+ T cells ~1 x 10 9 In some of the provided methods, the dose of engineered T cells is between 4.5 x 10 and 5 x 10 CAR+ T cells. 8 or approximately that number of CAR+ T cells.

[0038] In some of the provided methods, the method does not include determining the level of soluble BCMA in a tissue sample from the subject.

[0039] In some of the provided methods, the method does not include determining the level of interleukin-6 (IL-6) in a tissue sample from the subject.

[0040] In some of the provided methods, the method does not include determining the level of tumor necrosis factor alpha (TNFα) in a tissue sample from the subject.

[0041] In some of the provided methods, the subject has a serum M protein level of about 0.5 g / dL or greater or a urinary M protein level of about 200 mg / 24 hours or greater. In some of the provided methods, the R / R MM is light chain MM without measurable disease in the serum or urine. In some of the provided methods, the subject has a serum immunoglobulin free light chain level of about 10 mg / dL or greater or an abnormal serum immunoglobulin kappa lambda free light chain ratio.

[0042] In some of the provided methods, prior to the method, the subject has undergone one or more prior therapies to treat R / R MM. In some of the provided methods, the one or more prior therapies include an immunomodulatory agent. In some of the provided methods, the subject has undergone at least two consecutive cycles of an immunomodulatory agent.

[0043] In some of the provided methods, the subject has undergone between one and three prior therapies to treat R / R MM. In some of the provided methods, the subject has undergone at least three prior therapies to treat R / R MM.

[0044] In some of the provided methods, the one or more prior therapies include an anti-CD38 antibody and a proteasome inhibitor. In some of the provided methods, the subject has received at least two consecutive cycles of an anti-CD38 antibody and / or a proteasome inhibitor.

[0045] In some of any of the provided methods, the subject has relapsed or been refractory to one or more prior therapies.

[0046] In some of the provided methods, the subject exhibited progressive disease (PD) during or within six months of the end of one or more prior therapies. In some of the provided methods, the subject exhibited minimal response or better to at least one of the one or more prior therapies.

[0047] In some of the provided methods, the one or more prior therapies do not include an allogeneic hematopoietic stem cell transplant, a gene therapy-based therapeutic for cancer, an investigational cell therapy for cancer, or a BCMA-targeted therapy.

[0048] In some of the provided methods, the T cell therapy is prepared from a leukapheresis product and the subject has not undergone an autologous stem cell transplant (ASCT) within 12 weeks prior to leukapheresis. In some of the provided methods, the T cell therapy is prepared from a leukapheresis product and the subject has not undergone an autologous stem cell transplant (ASCT) within 12 months prior to leukapheresis.

[0049] In some of any of the provided methods, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

[0050] In some of any of the provided methods, the CAR comprises an antigen-binding domain that binds to BCMA, a transmembrane domain, and an intracellular signaling region comprising the CD3-zeta (CD3ζ) chain.

[0051] In some of the methods provided, the antigen-binding domain is a single-chain variable fragment (scFv).

[0052] In some of the provided methods, the CAR, e.g., the antigen binding domain, is H and V L Includes the area, V H The region includes CDR-H1 set forth in SEQ ID NO: 62, CDR-H2 set forth in SEQ ID NO: 63, and CDR-H3 set forth in SEQ ID NO: 64, L The region comprises a CDR-L1 set forth in SEQ ID NO: 65, a CDR-L2 set forth in SEQ ID NO: 66, and a CDR-H3 set forth in SEQ ID NO: 67. In some of any of the provided methods, the antigen-binding domain comprises a VH1 domain having a sequence of amino acids set forth in SEQ ID NO: 30, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30. H region, and V having a sequence of amino acids set forth in SEQ ID NO:31 or a sequence of amino acids exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:31. L In some of the provided methods, the antigen-binding domain comprises a V region having the sequence of amino acids set forth in SEQ ID NO: 30. Hregion and V having the amino acid sequence set forth in SEQ ID NO: 31 L Includes the area.

[0053] In some of the provided methods, the antigen-binding domain is an scFv having a sequence of amino acids set forth in SEQ ID NO: 68 or a sequence of amino acids exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 68. In some of the provided methods, the antigen-binding domain is an scFv set forth in SEQ ID NO: 68.

[0054] In some of the provided methods, the intracellular signaling region further comprises a costimulatory signaling region. In some of the provided methods, the costimulatory signaling region comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof. In some of the provided methods, the costimulatory signaling region comprises the intracellular signaling domain of 4-1BB, suitably human 4-1BB. In some of the provided methods, the costimulatory signaling region comprises the intracellular signaling domain of human 4-1BB. In some of the provided methods, the costimulatory signaling region is between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain.

[0055] In some of the provided methods, the transmembrane domain is or includes a transmembrane domain from human CD8.

[0056] In some of the provided methods, the CAR further comprises an extracellular spacer between the antigen-binding domain and the transmembrane domain. In some of the provided methods, the spacer is between at or about 50 amino acids and at or about 250 amino acids. In some of the provided methods, the spacer is a CD8 hinge.

[0057] In some of the provided methods, the CAR has a sequence set forth in any one of SEQ ID NOs: 126-177 or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 126-177. In some of the provided methods, the CAR has a sequence of amino acids set forth in SEQ ID NO: 152 or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 152.

[0058] In some of the provided methods, the T cell therapy includes idecbutagen-Biclucel cells. In some of the provided methods, the T cell therapy is ABECMA®.

[0059] In some of the provided methods, the T cell therapy includes siltacabtagene-autolucel cells. In some of the provided methods, the T cell therapy is CARVYKTI™.

[0060] Also provided is a therapy, e.g., a combination therapy, including one or more, e.g., all, of: a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds BCMA; an immunomodulatory compound that is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; and dexamethasone, for use in a method of treating multiple myeloma in accordance with any of the provided methods.

[0061] Also provided is the use of a therapy, e.g., a combination therapy, including one or more, e.g., all, of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds BCMA, an immunomodulatory compound that is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and dexamethasone, in a method of treating multiple myeloma in accordance with any of the provided methods.

[0062] Also provided is the use of a therapy, e.g., a combination therapy, including one or more, e.g., all, of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds BCMA, an immunomodulatory compound that is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, and dexamethasone, for the manufacture of a medicament for treating multiple myeloma in accordance with any of the provided methods.

[0063] In some of the provided embodiments, the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione. In some of the provided embodiments, the immunomodulatory compound is a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione.

[0064] In some of any of the provided embodiments, the dexamethasone is formulated for low dose administration.

[0065] Also provided is a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds BCMA, and a therapy comprising one or both, e.g., both, of a standard triplet regimen, e.g., a combination therapy, for use in a method of treating multiple myeloma in accordance with any of the provided methods.

[0066] Also provided is the use of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds BCMA and one or both of a standard triplet regimen, e.g., a therapy comprising both, e.g., a combination therapy, in a method of treating multiple myeloma in accordance with any of the provided methods.

[0067] Also provided is the use of a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds BCMA, and a therapy comprising one or both, e.g., both, of a standard triplet regimen, e.g., a combination therapy, for the manufacture of a medicament for treating multiple myeloma in accordance with any of the provided methods.

[0068] In some of the provided embodiments, the standard triplet regimen includes daratumumab, pomalidomide, and dexamethasone. In some of the provided embodiments, the standard triplet regimen includes pomalidomide, bortezomib, and dexamethasone.

[0069] In some of any of the provided embodiments, the dexamethasone is formulated for low dose administration. DETAILED DESCRIPTION OF THE INVENTION

[0070] Provided herein are combination therapies involving the administration of cellular immunotherapy via adoptive therapy of immune cells with cytotoxic activity, such as through T cell function or activity, including CAR-T cell therapy, in combination with one or more combination drugs for the treatment of a subject with cancer or a proliferative disease. In some embodiments, the cellular therapy, such as CAR T cell therapy, is a BCMA-targeted cellular therapy. In some embodiments, the combination therapy is used to treat multiple myeloma, including relapsed and refractory (R / R) multiple myeloma. The provided methods and combination therapies include methods and combination therapies involving certain immunomodulatory compounds and agents administered to enhance the activity of cells of the cellular therapy (e.g., CAR T cell therapy, such as BCMA-targeted CAR T cell therapy), such as to increase the activity and persistence, including durable activity, of the cells of the cellular therapy, following administration to a subject to treat cancer or a proliferative disease, such as multiple myeloma.

[0071] In some embodiments, the combination involves the administration of an immunomodulatory compound, such as a structural or functional analog of thalidomide and / or an inhibitor of E3-ubiquitin ligase, and the administration of a T cell therapy, such as a composition comprising cells (e.g., CAR-expressing T cells) for adoptive cell therapy, such as T cell therapy. In some embodiments, the combination involves the administration of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione [ka] or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, tautomer, or racemic mixture thereof (Compound A). (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione (Compound A) is a cereblon E3 ligase modulating compound (CELMoD). Compound A modulates CRBN, thereby inducing ubiquitination of the transcription factors Aiolos and Ikaros, increasing their proteasome-dependent degradation, and enhancing T cell function. Compound A binds to CRBN more potently than lenalidomide and pomalidomide, is more efficient at degrading Aiolos and Ikaros, and has a potent direct antiproliferative effect in lymphoma cells. In some embodiments, the combination drug further comprises dexamethasone. In some embodiments, the combination agent comprises Compound A and dexamethasone. In any of the provided methods, the dexamethasone is administered at a low dose.

[0072] In some embodiments, the combination agents include agents given as part of a standard triplet regimen, e.g., a triplet regimen that is the standard for treating cancer or proliferative diseases, such as multiple myeloma. In some embodiments, the combination agents are administered as part of or according to a standard triplet regimen. In some embodiments, the cancer or proliferative disease is multiple myeloma (MM), e.g., relapsed or refractory MM (R / R MM). In some embodiments, the combination agents include agents administered as part of a triplet regimen that is the standard for treating MM, e.g., R / R MM. In some embodiments, for example, the combination agents for a standard triplet regimen include daratumumab, pomalidomide, and dexamethasone. In some embodiments, for example, the combination agents for a standard triplet regimen include pomalidomide, bortezomib, and dexamethasone. In any of the provided methods, the dexamethasone is administered at a low dose.

[0073] Among the provided embodiments, particular mention is made of combination treatments involving BCMA and cellular immunotherapies, such as T cell therapies, that target or are directed against BCMA-expressing cells and diseases. BCMA is expressed in certain diseases and conditions, e.g., malignant lesions or tissues or cells thereof, e.g., in malignant plasma cells derived from any relapsed or newly diagnosed myeloma patient, and is observed, e.g., to be heterogeneously expressed, e.g., with little or no expression in normal tissues. Among the provided embodiments, particular mention is made of approaches useful in treating such diseases and conditions and / or for targeting such cell types, including by administering cellular immunotherapies containing nucleic acid molecules encoding BCMA-binding receptors, including chimeric antigen receptors (CARs), and immune cells, such as T cells or NK cells, engineered with the encoded receptors, such as the encoded CARs, and compositions and articles of manufacture comprising the same. Receptors, such as CARs, are generally composed of antibodies (including scFvs) specific for BCMA, heavy chain variable (V) antibodies. H ) region, including antigen-binding antibody fragments such as single domain antibody fragments and single chain fragments.

[0074] Among the diseases to be treated, any disease or disorder associated with BCMA, or any disease or disorder in which BCMA is specifically expressed and / or in which BCMA is targeted for treatment (also interchangeably referred to herein as "BCMA-associated disease or disorder"). Cancers associated with BCMA expression include hematological malignancies such as multiple myeloma, Waldenstrom's hypergammaglobulinemia, and both Hodgkin's and non-Hodgkin's lymphoma. For a review of BCMA, see Coquery et al., Crit Rev Immunol., 2012, 32(4):287-305. BCMA has been implicated in mediating tumor cell survival and is therefore a potential target for cancer therapy. Chimeric antigen receptors containing mouse anti-human BCMA antibodies and cells expressing such chimeric receptors have previously been described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060.

[0075] In some embodiments, the provided methods enhance or modulate the proliferation and / or activity of T cell activity associated with the administration of a cellular immunotherapy, such as a composition comprising cells (e.g., CAR-expressing T cells) for adoptive cell therapy, such as T cell therapy.

[0076] In some embodiments, the combination therapy involves administration of dexamethasone. In some embodiments, dexamethasone is administered in combination with an immunomodulatory compound (e.g., Compound A) and T cell therapy (e.g., anti-BCMA CAR T cells). In some embodiments, dexamethasone is administered as part of a standard triplet regimen administered in combination with T cell therapy (e.g., anti-BCMA CAR T cells). In some embodiments, the standard triplet regimen is a DPd regimen (e.g., including daratumumab, pomalidomide, and dexamethasone). In some embodiments, the standard triplet regimen is a PVd regimen (e.g., including pomalidomide, bortezomib, and dexamethasone).

[0077] T cell-based therapies, e.g., adoptive T cell therapies (including therapies involving the administration of cells expressing chimeric receptors specific for a disease or disorder of interest, such as chimeric antigen receptors (CARs) and / or other recombinant antigen receptors, as well as other adoptive immune cell and adoptive T cell therapies), can be effective in treating cancer and other diseases and disorders. The engineered expression of recombinant receptors, such as chimeric antigen receptors (CARs), on the surface of T cells allows for the redirection of T cell specificity.

[0078] In certain situations, available approaches to adoptive cell therapy may not always be entirely satisfactory. For example, in certain cases, while CAR T cell persistence can be detected in many subjects, the response in some subjects is transient, and the subject has been shown to relapse in the presence of persistent CAR T cells.

[0079] In some embodiments, this may be explained by the immunological exhaustion of circulating CAR-expressing T cells and / or changes in T lymphocyte populations. In some situations, optimal efficacy may depend on the ability of the administered cells to recognize and bind to a target, such as a target antigen, transport to the appropriate site within the subject, tumor, and its environment, localize there, and successfully enter there. In some situations, optimal efficacy may depend on the ability of the administered cells to become activated, proliferate, exert various effector functions, including cytotoxic killing and secretion of various factors such as cytokines, persist, including long-term, differentiate, transition to, or engage in reprogramming of a specific phenotypic state (such as long-lived memory, low differentiation, and effector state), avoid or reduce the immunosuppressive state in the local microenvironment of the disease, produce an effective and robust recall response after clearance and re-exposure to the target ligand or antigen, and avoid or reduce exhaustion, anergy, peripheral tolerance, terminal differentiation, and / or differentiation into an inhibitory state.

[0080] In some embodiments, the exposure and persistence of the engineered cells is reduced or attenuated after administration to a subject. Nevertheless, observations indicate that in some cases, increasing a subject's exposure to administered cells expressing the recombinant receptor (e.g., increasing the number or duration of cells over time) can improve the efficacy and therapeutic outcome of adoptive cell therapy.

[0081] In some embodiments, after prolonged stimulation or exposure to antigen and / or exposure under conditions in the tumor microenvironment, T cells can become hypofunctional and / or exhibit characteristics associated with an exhausted state over time. In some aspects, this reduces the persistence and efficacy of the T cells against the antigen and limits their ability to be effective. In particular, there is a need for methods to improve the efficacy and function of CART cells to minimize, reduce, prevent, or reverse the hypofunctional or exhausted state.

[0082] Compound A has been shown to directly affect malignant lymphocyte survival through the degradation of Ikaros family transcription factors. The molecular target of Compound A was identified as the protein cereblon (CRBN), a substrate receptor for the Cullin4 RING E3 ubiquitin ligase complex. Binding to the hydrophobic tryptophan pocket within CRBN promotes the recruitment, ubiquitination, and subsequent proteasomal degradation of several protein substrates, including Aiolos (IKZF3) and Ikaros (IKZF1). Ikaros is expressed in the immature stage of myeloid differentiation and regulates early neutrophil differentiation (Dumortier et al. (2003) Blood 101:2219). Thus, in some cases, depletion of Ikaros, such as by administering an immunomodulatory compound, e.g., Compound A, to a subject, can result in neutropenia in some instances. In some embodiments, providing the combination therapy reduces or mitigates the risk of toxicity, such as neutropenia and other toxicities, including cytokine release syndrome (CRS) or neurotoxicity, as a result of the combination of cell therapy and Compound A.

[0083] In addition to their cell-autonomous activity against malignant B cells, E3 ligase-modulating compounds such as Compound A also exert costimulatory effects on immune cells such as T and NK cells. This activity has also been shown to be mediated by CRBN-mediated degradation of activation molecules and cytokines such as Aiolos and Ikaros, negative regulators of interleukin-2 (IL-2) expression (Gandhi, Br J Haematol. 2014 Mar;164(6):811-21, Kroenke, Oncoimmunology, 2014;3(7):e941742.).

[0084] In some embodiments, the combination agents administered as part of the provided methods can improve T cell function, including functions related to the ability of T cells to produce one or more cytokines, cytotoxicity, expansion, proliferation, and persistence. In some embodiments, the provided methods enhance or modulate the proliferation and / or activity of T cells (e.g., CAR-expressing T cells) associated with the administration of T cell therapy. In some embodiments, the provided methods achieve improved or better T cell functionality, and thereby improved anti-tumor efficacy.

[0085] In some aspects, the combination agents administered as part of the provided methods can be effective in reversing, delaying, or preventing T cell exhaustion, including by increasing T cell signaling and / or altering one or more genes that are differentially regulated following chronic (long-term) stimulation. Thus, while in some cases, agents that increase or enhance T cell activity may drive cells into an exhausted state, in some aspects, the activity of the combination agents administered as part of the provided methods to exert an enhancing effect on T cell activity can be uncoupled from T cell exhaustion. In some embodiments, the provided methods involving administration of such combination agents can enhance the activity of naive T cells and delay, limit, reduce, inhibit, or prevent exhaustion. In some aspects, the provided methods can also achieve improved or more durable responses, such as in certain groups of treated subjects, compared to certain alternative methods.

[0086] In some aspects, the combination agents administered as part of the provided methods can promote the continued function and / or survival of T cell therapy (e.g., CAR-T cell) cells after activation, including after antigen encounter. In some aspects, the combination agents administered as part of the provided methods can increase the ability of such T cells to persist or function long-term, such as by preventing exhaustion or cell death. In certain embodiments, the combination therapy described herein can provide a useful therapeutic approach for enhancing and prolonging the activity of CAR T cells across B-cell malignancies by modulating the tumor microenvironment by improving the sustained anti-tumor function of CAR T cells. In some cases, the combination agents administered as part of the methods provided herein can also have a direct anti-tumor effect in lymphoma cells. In some embodiments, such improvement can result in the combination therapy exhibiting an improved overall response, e.g., reduced tumor burden and / or increased survival, compared to subjects treated with monotherapy involving administration of the T cell therapy (e.g., CAR-T cell) or combination agent alone. In some embodiments, the provided methods can increase overall response and / or survival by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold or more compared to alternative treatments, such as compared to monotherapy involving administration of T cell therapy (e.g., CAR-T cells) or a combination agent alone.

[0087] The provided methods include administering a combination agent in an effective amount that exhibits a T cell modulating effect. In some embodiments, exemplary combination agents at particular dosages can increase or enhance T cell function in T cell therapy, e.g., CAR-T cell therapy.

[0088] In some embodiments, the combination with a combination agent improves one or more outcomes or functional attributes, but does not affect one or more side effects or undesired changes in the T cells, such as not reducing the ability of the cells to become activated, secrete one or more desired cytokines, expand, and / or persist, e.g., as measured in in vitro assays compared to such cells cultured under otherwise identical conditions but in the absence of the combination agent. Thus, in some embodiments, methods and combinations are provided that result in improvements in T cell function or phenotype, e.g., intrinsic T cell functionality and / or intrinsic T cell phenotype, generally without compromising one or more other desired properties of functionality, e.g., CAR-T cell functionality.

[0089] In some embodiments, the provided methods can enhance T cell therapy, e.g., CAR-T cell therapy, which can, in some aspects, improve the outcome of treatment. In some embodiments, the methods are particularly advantageous in subjects in which the cells of the T cell therapy show weak expansion, become exhausted, show reduced or decreased persistence in the subject, and / or in subjects with cancer that is resistant or refractory to other therapies, has high-grade or high-risk cancer, and / or has or may show a relatively lower response rate to CAR-T cell therapy administered without a combination agent compared to another type of cancer or compared to administration with a different CAR-T cell therapy.

[0090] In some embodiments, the provided methods are used at a time point when T cell therapy (e.g., CAR T cells) may or may be exhibiting characteristics of exhaustion. In some embodiments, the exhaustion phenotype is evident after T cells have reached peak expansion and begin to decline in number in the subject's blood. In some embodiments, administration of the combination agent is performed at a time point when the T cells exhibit increased reduced function or exhaustion compared to a time point immediately prior to exposure of the T cells to an antigen (baseline) or compared to a time point when the cells have been exposed to an antigen but continue to proliferate and have not yet reached peak expansion. In some embodiments, the increased reduced function or exhaustion can be determined by increased expression of exhaustion markers compared to a previous, earlier time point. In some embodiments, the increased reduced function or exhaustion, such as increased expression of exhaustion markers, occurs following administration of T cell therapy (e.g., CAR T cells) to a subject with a disease or condition associated with the antigen targeted by the T cell therapy. T cells, such as T cells in the peripheral blood after administration to the subject, can be monitored for markers of T cell activation or exhaustion, such as PD-1, TIM-3, and LAG-3.

[0091] In some embodiments, a subject receiving T cell therapy, e.g., CAR-T cells, is monitored for the presence, absence, or level of therapeutic T cells in the subject, e.g., in the subject's biological sample, e.g., in the subject's blood. In some embodiments, a combination agent is administered to a subject who has received T cell therapy (e.g., CAR-T cells), but where such cells have weakly expanded and / or are at or below a threshold level in the subject's sample, e.g., blood sample, at a time when a strong or robust expansion of CAR-T cells in the subject is typically observed in multiple subjects who have received T cell therapy (e.g., CAR-T), and in some cases, the same T cell therapy (e.g., the same CAR-T cells).

[0092] In certain embodiments, the provided methods can enhance, increase, or potentiate T cell therapy in a subject in which a peak response to T cell therapy was observed, but the response, e.g., reduction in T cell presence and / or tumor burden, has become reduced or is no longer detectable. In some embodiments, the combination agent is capable of: (i) causing a peak or maximum level of T cell therapy cells to be detectable in the subject's blood; (ii) causing the number of T cell therapy cells detectable in the blood to become undetectable or reduced, optionally compared to a prior time point after administration of the T cell therapy; (iii) causing the number of T cell therapy cells detectable in the blood to be reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more, relative to the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of administration of the T cell therapy; (iv) causing a T cell therapy that is more potent than the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of administration of the T cell therapy; (v) the subject exhibits disease progression and / or relapse following remission after treatment with T cell therapy; and / or (iv) the subject exhibits an increased tumor burden compared to the tumor burden before or after administration of the T cells and before initiation of administration of the combination agent.

[0093] In some embodiments, the methods can be used to treat a disease or condition, e.g., multiple myeloma, e.g., relapsed / refractory multiple myeloma. In some embodiments, the methods can be used to treat such diseases, conditions, or malignancies in which the response, e.g., complete response, to treatment with T cell therapy alone, e.g., a composition comprising cells (e.g., CAR-expressing T cells) for adoptive cell therapy, such as T cell therapy, is relatively low (e.g., CR in less than or about 60%, less than about 50%, or less than about 45% of subjects so treated), compared to treatment with other T cell therapies or treatments of other diseases or malignancies, and / or the subject is not responsive to treatment with the combination agent alone.

[0094] In some embodiments, the combination therapies provided herein are for use in subjects with cancer in which, after initiation of administration of T cell therapy, such as a composition comprising cells for adoptive cell therapy, e.g., CAR-expressing T cells, the subject has relapsed following remission after treatment with T cell therapy. In some embodiments, subjects who have relapsed following such remission are administered a combination agent. In some embodiments, the combination therapies provided herein are for use in subjects with a disease or condition, e.g., cancer, in which the amount of the administered combination agent, as a single agent and / or in the absence of administration of T cell therapy, is insufficient to ameliorate, reduce, or prevent the disease or condition, or its symptoms or outcomes, e.g., is insufficient to ameliorate, reduce, or prevent the disease or condition, or its symptoms or outcomes, in the subject. In some embodiments, the method thereby reduces or ameliorates the symptoms or outcome or burden of a disease or condition to an extent that exceeds the combination of (i) the degree of reduction or amelioration, on average in a population of subjects with the disease or condition, that results from the administration of an immunomodulatory agent alone, and (ii) the degree of reduction or amelioration, on average in a population of subjects with the disease or condition, that results from the administration of T cell therapy alone, as appropriate. In some embodiments, the method reduces or ameliorate such symptoms, outcome or burden of the disease by more than or about 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 6.0-fold, 7.0-fold, 8.0-fold, 9.0-fold, 10.0-fold, 20.0-fold, 30.0-fold, 40.0-fold, 50.0-fold or more, e.g., compared to that in a population of subjects with the disease or condition, on average.

[0095] In some embodiments of the provided methods, one or more characteristics of the administered engineered cells may be improved, increased, or superior compared to the administered cells of a reference composition, e.g., increased or longer expansion and / or persistence of such administered cells in a subject, or increased or greater recall response after restimulation with an antigen. In some embodiments, the increase may be at least 1.2-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, or at least 10-fold increase in such a characteristic or trait compared to the same characteristic or trait after administration of a reference cell composition. In some embodiments, an increase in one or more of such characteristics or traits can be observed or is present within 7, 14, 21 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or 12 months after initiation of administration of the engineered cells and the combination agent.

[0096] In some embodiments, the combination agent is administered to the subject a sufficient time after receiving lymphodepleting therapy so that any myelosuppressive effects of the combination agent and lymphodepleting therapy are minimized.

[0097] In some embodiments, the provided methods do not result in a high rate or likelihood of toxicity or a toxic outcome, or reduce the rate or likelihood of toxicity or a toxic outcome, e.g., neurotoxicity (NT), cytokine release syndrome (CRS), or hematological toxicity, e.g., neutropenia, such as compared to certain other cell therapy or immunomodulatory regimens.

[0098] In some embodiments, the method does not result in or increase the risk of certain hematological toxicities, such as neutropenia or thrombocytopenia. In some embodiments, 50% or less of the subjects exhibit neutropenia greater than Grade 3, e.g., prolonged Grade 3 or Grade 4 neutropenia, and / or thrombocytopenia greater than Grade 3, e.g., Grade 3 or Grade 4 thrombocytopenia. In some embodiments, at least 50% of the subjects treated according to the method (e.g., at least 60%, at least 70%, at least 80%, at least 90% or more of the treated subjects) do not exhibit severe neutropenia or severe thrombocytopenia of Grade 3 or greater than Grade 3.

[0099] In some embodiments, the methods do not result in or increase the risk of severe NT (sNT), severe CRS (sCRS), macrophage activation syndrome, tumor lysis syndrome, fever of at least or about 38 degrees Celsius for three or more days, and plasma levels of CRP of at least or about 20 mg / dL. In some embodiments, more than or about 30%, 35%, 40%, 50%, 55%, 60% or more of subjects treated according to the provided methods do not exhibit any grade of CRS or any grade of neurotoxicity. In some embodiments, 50% or less of treated subjects (e.g., at least 60%, at least 70%, at least 80%, at least 90% or more of treated subjects) exhibit cytokine release syndrome (CRS) greater than Grade 2 and / or neurotoxicity greater than Grade 2. In some embodiments, at least 50% of the subjects treated according to the method (e.g., at least 60%, at least 70%, at least 80%, at least 90% or more of the treated subjects) do not experience a severe toxic outcome (e.g., severe CRS or severe neurotoxicity), e.g., do not experience grade 3 or higher neurotoxicity, and / or do not experience severe CRS, or do not do so within a certain period of time following treatment, e.g., within 1 week, 2 weeks, or 1 month of administration of the cells.

[0100] In some cases, the combination agent is administered at a time when it can efficiently / effectively boost or prime the cells. In some embodiments, administration of the combination agent is initiated at or before the peak or maximum level of the cell therapy cells is detectable in the subject's blood. In some embodiments, the provided method can enhance T cell therapy, e.g., CAR-T cell therapy, which can, in some aspects, improve the outcome of treatment. In some embodiments, the method is particularly advantageous in subjects in which the T cell therapy cells show weak expansion, become exhausted, show reduced or decreased persistence in the subject, and / or in subjects with cancer that is resistant or refractory to other therapies and / or is a high-grade or high-risk cancer.

[0101] In some embodiments, a subject receiving T cell therapy, e.g., CAR-T cells, is monitored for the presence, absence, or level of therapeutic T cells in the subject, e.g., in the subject's biological sample, e.g., in the subject's blood. In some embodiments, the provided methods result in engineered cells with increased persistence and / or better efficacy in the subject to which they are administered. In some embodiments, the persistence of engineered cells, e.g., CAR-expressing T cells, in the subject is greater than the persistence that would be achieved by alternative methods, such as those involving administration of T cell therapy but without administration of a combination agent. In some embodiments, the persistence is increased by at least or about at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more.

[0102] In some embodiments, the degree or level of persistence of administered cells can be detected or quantified after administration to a subject. For example, in some aspects, quantitative PCR (qPCR) is used to assess the content of cells expressing the recombinant receptor (e.g., CAR-expressing cells) in the subject's blood or serum or organs or tissues (e.g., disease site). In some aspects, persistence is quantified as copies of the DNA or plasmid encoding the receptor, e.g., CAR, per microgram of DNA, or as the number of receptor-expressing, e.g., CAR-expressing, cells per microliter of sample, e.g., blood or serum, or per microliter of sample, or per total number of peripheral blood mononuclear cells (PBMCs) or white blood cells or T cells. In some embodiments, flow cytometry assays can also be performed to detect receptor-expressing cells, generally using antibodies specific to the receptor. Cell-based assays can also be used to detect the number or percentage of functional cells, such as cells that can bind to and / or neutralize and / or respond to, e.g., induce a cytotoxic response, disease or condition cells or cells expressing an antigen recognized by the receptor. In any of such embodiments, the degree or level of expression of another marker associated with the recombinant receptor (e.g., CAR-expressing cells) can be used to distinguish the administered cells from endogenous cells in the subject.

[0103] In some embodiments, the combination agent is administered for a period that enhances, increases, or optimizes the durability of the response. In some aspects, the provided methods are based on the observation that subjects who achieve or are likely to achieve complete remission (CR) at 3 months, e.g., generally 6 months, are more likely to sustain the response for a longer period, e.g., survive or survive without progression for more than or about 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months after initially achieving complete response (CR) following administration of the combination therapy. In some aspects, the method is carried out to administer the combination agent for a period that is at least 3 months, e.g., at least 4 months, at least 5 months, or at least 6 months after initiating administration of T cell therapy. In some embodiments, the combination agent is administered for at least 6 months or at least 180 days after initiating administration of T cell therapy. In some embodiments, if the subject shows CR at the end of the period, or if the disease or condition progresses or recurs in the subject after receiving treatment (combination therapy), the administration of the combination agent will be terminated or stopped. In some aspects, continued administration of the combination agent can be carried out in subjects who show partial response (PR) or stable disease (SD) at the end of the period (e.g., at or about 6 months). In other aspects, the period is a fixed duration, and further administration of the combination agent is not carried out.

[0104] In some aspects, the provided methods and uses provide or achieve improved or more durable responses or efficacy compared to certain alternative methods, e.g., methods that involve administration of T cell therapy or combinations as monotherapy, or methods that do not involve administration as combined combination therapy as described herein, such as in certain groups of treated subjects. In some embodiments, the methods are advantageously based on the administration of T cell therapy, e.g., a composition comprising cells (e.g., CAR-expressing T cells) for adoptive cell therapy, such as T cell therapy, and a combination agent. In some embodiments, such responses are observed in high-risk patients with poor prognosis, such as multiple myeloma that is relapsed or refractory (R / R) to standard therapy or has a poor prognosis.

[0105] In some embodiments, at least 35%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture, kits, or compositions achieve a complete response (CR). In some embodiments, the subjects are in CR and exhibit minimal residual disease (MRD). In some embodiments, the subjects are in CR and are MRD-. In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of subjects treated according to the provided methods and / or with the provided articles of manufacture, kits, or compositions achieve an objective response of partial response (PR). In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or more of subjects treated according to the provided methods and / or using the provided articles of manufacture, kits, or compositions achieve a CR or PR at 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 1 year after initiation of administration of the cell therapy.

[0106] In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or more of subjects treated according to the provided methods and / or using the provided articles of manufacture, kits, or compositions maintain their response, e.g., maintain a CR or objective response (OR), by 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months or more after initiation of cell therapy administration. In some embodiments, such a response, e.g., a CR or OR, is durable for at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more, such as in at least or about at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or more of subjects treated according to the provided methods, or in such subjects achieving a CR by 3, 4, 5, or 6 months. In some embodiments, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or more of subjects treated according to the provided methods and / or using the provided articles of manufacture, kits, or compositions, or those subjects who achieve a CR by 3, 4, 5, or 6 months, survive or survive progression-free for more than or about 6, 7, 8, 9, 10, 11, 12, or more months.

[0107] In some embodiments, the reference cell composition may be a composition of T cells derived from the blood of a subject who does not have or is not suspected of having cancer, or a population of T cells obtained, isolated, generated, produced, incubated, and / or administered under the same or substantially the same conditions, except that they are not incubated or administered in the presence of a combination drug. In some embodiments, the reference cell composition contains substantially the same genetically engineered cells, including the expression of the same recombinant receptor, e.g., CAR. In some aspects, such T cells are treated identically or substantially the same, e.g., similarly manufactured, similarly formulated, administered with the same or approximately the same dosage and other similar factors.

[0108] In some embodiments, the provided methods result in genetically engineered cells that have increased persistence and / or better efficacy in the subject to which they are administered. In some embodiments, the persistence of the genetically engineered cells, e.g., CAR-expressing T cells, in the subject is greater than the persistence that would be achieved by an alternative method, such as administering a reference cell composition, e.g., a T cell therapy, but in the absence of administering a combination agent. In some embodiments, the persistence is increased by at least or about at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more.

[0109] In some embodiments, the degree or level of persistence of administered cells can be detected or quantified after administration to a subject. For example, in some aspects, quantitative PCR (qPCR) is used to assess the content of cells expressing the recombinant receptor (e.g., CAR-expressing cells) in the subject's blood or serum or organs or tissues (e.g., disease site). In some aspects, persistence is quantified as copies of the DNA or plasmid encoding the receptor, e.g., CAR, per microgram of DNA, or as the number of receptor-expressing, e.g., CAR-expressing, cells per microliter of sample, e.g., blood or serum, or per microliter of sample, or per total number of peripheral blood mononuclear cells (PBMCs) or white blood cells or T cells. In some embodiments, flow cytometry assays can also be performed to detect receptor-expressing cells, generally using antibodies specific to the receptor. Cell-based assays can also be used to detect the number or percentage of functional cells, such as cells that can bind to and / or neutralize and / or respond to, e.g., induce a cytotoxic response, disease or condition cells or cells expressing an antigen recognized by the receptor. In any of such embodiments, the degree or level of expression of another marker associated with the recombinant receptor (e.g., CAR-expressing cells) can be used to distinguish the administered cells from endogenous cells in the subject.

[0110] Also provided are methods for manipulating, preparing, and producing the cells, compositions containing the cells and / or combination agents, and kits and devices containing, using, producing, and administering the cells and / or combination agents, such as for the provided methods of combination therapy.

[0111] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated herein by reference. To the extent that a definition set forth herein contradicts or is otherwise inconsistent with a definition set forth in a patent, application, published application, or other publication incorporated herein by reference, the definition set forth herein takes precedence over the definition incorporated herein by reference.

[0112] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. I. Combination Therapy

[0113] Methods and uses of engineered cells, e.g., T cells (e.g., CAR-T cells), in combination with one or more combination agents for the treatment of a subject with cancer are provided. In some embodiments, the combination agent comprises an immunomodulatory compound. In some embodiments, the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or a compound represented by Formula I [ka] or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, tautomer, or racemic mixture thereof (Compound A). In some embodiments, the combination drug includes dexamethasone. In some embodiments, the combination drug includes an immunomodulatory agent, such as Compound A, and dexamethasone. In some embodiments, the combination drug includes agents given as part of a standard triplet regimen for the treatment of cancer. In some embodiments, the combination drug is administered as part of a standard triplet regimen for the treatment of cancer. In some embodiments, the combination drug includes daratumumab, pomalidomide, and dexamethasone. In some embodiments, the combination drug includes pomalidomide, bortezomib, and dexamethasone. In certain embodiments, the method is for treating multiple myeloma. In some embodiments, the multiple myeloma is relapsed or refractory multiple myeloma.

[0114] In some embodiments, the cell therapy is adoptive cell therapy. In some embodiments, the cell therapy is or includes a recombinant receptor-expressing cell therapy (optionally T cell therapy), which is tumor-infiltrating lymphocyte (TIL) therapy, transgenic TCR therapy, or optionally chimeric antigen receptor (CAR)-expressing cell therapy. In some embodiments, the therapy is a B cell targeted therapy. In some embodiments, the therapy targets B cell maturation antigen (BCMA). In some embodiments, the cells and dosage regimens for administering the cells can include any described in Section IA.

[0115] In some embodiments, dosage regimens for administering immunomodulatory compounds can include any of those described in Section IB-2.

[0116] In some embodiments, dosage regimens for administering dexamethasone, including dexamethasone, in combination with an immunomodulatory compound and cell therapy can include any described in Section IB-3.

[0117] In some embodiments, dosage regimens for administering the standard triplet regimen can include any of those described in Section IB-4.

[0118] In some embodiments, T cell therapy (e.g., CAR-expressing T cells) and combination agents are provided as pharmaceutical compositions for administration to a subject. In some embodiments, the pharmaceutical composition contains a therapeutically effective amount of one or both of the T cells for cell therapy and combination agents described. In some embodiments, the T cells for cell therapy and combination agents are formulated for administration in separate pharmaceutical compositions. In some embodiments, any of the pharmaceutical compositions provided herein can be formulated in a dosage form appropriate for each administration route.

[0119] In some embodiments, a combination therapy comprising administering a T cell therapy comprising engineered cells, such as CAR-T cell therapy, and a combination agent is administered to a subject or patient having or at risk of having a disease or condition (e.g., cancer) to be treated. In some aspects, the method treats, e.g., ameliorates, one or more symptoms of the disease or condition, such as by reducing tumor burden in cancers that express an antigen recognized by the immunotherapy or immunotherapeutic agent, e.g., recognized by the engineered T cells.

[0120] In some embodiments, the disease or condition being treated may be one in which expression of an antigen is associated with and / or involved in the etiology of the disease, condition, or disorder, e.g., causes, exacerbates, or is otherwise involved in such disease, condition, or disorder. Exemplary diseases and conditions may include diseases or conditions associated with cellular malignancies or cancerous transformation (e.g., cancer), autoimmune or inflammatory diseases, or infectious diseases caused, for example, by bacteria, viruses, or other pathogens. Exemplary antigens, including antigens associated with various diseases and conditions that can be treated, include any of the antigens described herein. In certain embodiments, a recombinant receptor expressed in the engineered cells of the combination therapy, including a chimeric antigen receptor or transgenic TCR, specifically binds to an antigen associated with the disease or condition.

[0121] In some embodiments, the cancer or proliferative disease expresses BCMA. In some embodiments, the provided methods use recombinant receptor-expressing T cells (e.g., CAR-T cells) that target BCMA.

[0122] In certain embodiments, the anti-BCMA CAR T cell therapy is idecbutagen-bicrueucel, ABECMA©, BCMA02, JCARH125, JNJ-68284528 (LCAR-B38M; siltacbutagen-bicrueucel; CARVYKTI™) (Janssen / Legend), P-BCMA-101 (Poseida), PBCAR269A (Poseida), P-BCMA-Allo1 (Poseida), Allo-715 (Pfizer / Allogene), CT053 (Carsgen), Descartes-08 (Cartesian), PHE885 (Novartis), ARI-002 (Hospital Clinic Barcelona, ​​IDIBAPS), or CTX120 (CRISPR Therapeutics). In certain embodiments, the anti-BCMA CAR T cells are idecbutagen-bicrueucel cells. In certain embodiments, the anti-BCMA CAR T cells are ABECMA® cells (cells used in ABECMA® immunotherapy). In certain embodiments, the anti-BCMA CAR T cells are siltacabtagene-autolucel cells. In certain embodiments, the anti-BCMA CAR T cells are CARVYKTI™ cells (cells used in CARVYKTI™ immunotherapy).

[0123] In some embodiments, the methods and uses include: 1) administering to a subject a T cell therapy involving T cells expressing an engineered cell surface receptor (e.g., a recombinant antigen receptor), typically a chimeric receptor such as a chimeric antigen receptor (CAR) directed against or targeting BCMA; and 2) administering to a subject one or more combination agents. In some embodiments, administration of the combination agent begins after (following) administration of the T cell therapy or after (following) initiation of administration of the T cell therapy. In some cases, the combination agent is administered to a subject who has received T cell therapy. The methods generally involve administering to a subject one or more doses of cells and two or more doses of the combination agent.

[0124] Combination therapies comprising engineered cells expressing a recombinant receptor, such as a chimeric antigen receptor (CAR), as described herein, and a combination agent, or a composition comprising the engineered cells and / or the combination agent, are useful in a variety of therapeutic, diagnostic, and prophylactic applications. For example, the combinations are useful in treating various diseases and disorders in a subject. Such methods and uses include methods of therapy involving the administration of the engineered cells and the combination agent, and / or a composition containing one or both, to a subject having a disease, condition, or disorder, such as a tumor or cancer. In some embodiments, the engineered cells and the combination agent, and / or a composition containing one or both, are administered in an effective amount to provide treatment for the disease or disorder. Uses include the use of the engineered cells and the combination agent, and / or a composition containing one or both, in such methods and treatments and in the preparation of a medicament for carrying out such methods of therapy. In some embodiments, the methods are carried out by administering the engineered cells and the combination agent, and / or a composition containing one or both, to a subject having or suspected of having a disease or condition. In some embodiments, the method thereby treats the disease, condition, or disorder in the subject.

[0125] Among the diseases to be treated, any disease or disorder associated with BCMA, or any disease or disorder in which BCMA is specifically expressed and / or in which BCMA is targeted for treatment (also interchangeably referred to herein as "BCMA-associated disease or disorder"). Cancers associated with BCMA expression include hematological malignancies such as multiple myeloma, Waldenstrom's hypergammaglobulinemia, and both Hodgkin's and non-Hodgkin's lymphoma. For a review of BCMA, see Coquery et al., Crit Rev Immunol., 2012, 32(4):287-305. BCMA has been implicated in mediating tumor cell survival and is therefore a potential target for cancer therapy. Chimeric antigen receptors containing mouse anti-human BCMA antibodies and cells expressing such chimeric receptors have previously been described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060.

[0126] In some embodiments, the BCMA-associated disease or disorder is a B-cell-associated disorder, hi some embodiments, the BCMA-associated disease or disorder is one or more diseases or conditions selected from glioblastoma, lymphomatoid granulomatosis, post-transplant lymphoproliferative disorder, immunoregulatory disorder, heavy chain disease, primary or immune cell-associated amyloidosis, or monoclonal gammopathy of undetermined significance.

[0127] In some embodiments, the BCMA-associated disease or disorder is an autoimmune disease or disorder, including, but not limited to, systemic lupus erythematosus (SLE), lupus nephritis, inflammatory bowel disease, rheumatoid arthritis (e.g., juvenile rheumatoid arthritis), ANCA-associated vasculitis, idiopathic thrombocytopenic purpura (ITP), thrombotic thrombocytopenic purpura (TTP), autoimmune thrombocytopenia, Chagas' disease, Graves' disease, Wegener's granulomatosis, polyarteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, psoriasis, IgA nephropathy, IgM polyneuropathy, vasculitis, diabetes mellitus, Raynaud's syndrome, antiphospholipid syndrome, Goodpasture's disease, Kawasaki's disease, autoimmune hemolytic anemia, myasthenia gravis, or progressive glomerulonephritis.

[0128] Among the diseases, disorders, or conditions associated with BCMA, particular mention is made of cancer (e.g., BCMA-expressing cancers). Cancers that can be treated, e.g., BCMA-expressing cancers, include, but are not limited to, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma (e.g., multiple myeloma), gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer, and head and neck cancer.

[0129] In certain diseases and conditions, BCMA is expressed in malignant cells and cancers. In some embodiments, the cancer (e.g., a BCMA-expressing cancer) is a B-cell malignancy. In some embodiments, the cancer (e.g., a BCMA-expressing cancer) is a lymphoma, leukemia, or plasma cell malignancy. Lymphomas contemplated herein include, but are not limited to, Burkitt lymphoma (e.g., endemic Burkitt lymphoma or sporadic Burkitt lymphoma), non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, Waldenstrom's hypergammaglobulinemia, follicular lymphoma, small non-cleaved cell lymphoma, mucosa-associated lymphoid tissue lymphoma (MALT), marginal zone lymphoma, splenic lymphoma, nodular monocytoid B-cell lymphoma, immunoblastic lymphoma, large cell lymphoma, diffuse mixed cell lymphoma, pulmonary B-cell angiocentric lymphoma, small lymphocytic lymphoma, primary mediastinal B-cell lymphoma, lymphoplasmacytic lymphoma (LPL), or mantle cell lymphoma (MCL). Leukemias contemplated herein include, but are not limited to, chronic lymphocytic leukemia (CLL), plasma cell leukemia, or acute lymphocytic leukemia (ALL). Plasma cell malignancies, including, but not limited to, multiple myeloma (e.g., non-secretory multiple myeloma, smoldering multiple myeloma) or plasmacytoma, are also contemplated herein.

[0130] In some embodiments, the disease or condition is a plasmacytoma, such as an extramedullary plasmacytoma. In some embodiments, the subject does not have a plasmacytoma, such as an extramedullary plasmacytoma.

[0131] In some embodiments, the disease or condition is multiple myeloma (MM), such as relapsed and / or refractory multiple myeloma (R / R MM).

[0132] In some embodiments, the methods can identify subjects who have, are suspected of having, or are at risk of developing a BCMA-related disease or disorder. Thus, methods are provided for identifying subjects who have a disease or disorder associated with elevated BCMA expression and selecting the subject for treatment with BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells).

[0133] In some aspects, for example, a subject can be screened for the presence of a disease or disorder associated with elevated BCMA expression, such as a BCMA-expressing cancer. In some embodiments, the method includes screening for or detecting the presence of a BCMA-associated disease, e.g., a tumor or cancer, e.g., multiple myeloma. Thus, in some aspects, a sample can be obtained from a patient suspected of having a disease or disorder associated with elevated BCMA expression and assayed for BCMA expression levels. In some aspects, a subject who tests positive for a BCMA-associated disease or disorder can be selected for treatment by the method and administered a therapeutically effective amount of a BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells) or pharmaceutical composition thereof described herein.

[0134] In some embodiments, a subject can be screened for the level of soluble BCMA (sBCMA) from a biological sample from the subject, such as, for example, blood or serum. In some embodiments, a subject can be screened for the level of sBCMA prior to treatment with a cell therapy. In some embodiments, a method includes screening for or detecting the level or amount of sBCMA in a subject with a disease or disorder associated with BCMA expression, e.g., a tumor or cancer, e.g., multiple myeloma. In some embodiments, a sample can be obtained from a patient suspected of having a BCMA-associated disease or disorder and assayed for the level or amount of sBCMA using an assay for detecting soluble protein levels, such as, for example, an enzyme-linked immunosorbent assay (ELISA). In some embodiments, in subjects with multiple myeloma (MM), sBCMA levels can be correlated with the percentage of plasma cells in a bone marrow biopsy. In some embodiments, in subjects with multiple myeloma (MM), sBCMA levels can correlate with a reduced response to treatment, or shorter overall or progression-free survival (see, e.g., Ghermezi et al., Haematologica 2017, 102(4): 785-795). In some embodiments, subjects exhibiting low sBCMA levels can be selected for treatment by the present methods and can be administered a therapeutically effective amount of a BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells) or pharmaceutical composition thereof described herein.

[0135] In some aspects, the method does not include screening a subject based on soluble BCMA levels. In some embodiments, the method does not include determining soluble BCMA levels in a sample from the subject. In some embodiments, the method does not include determining soluble BCMA levels in a tissue sample from the subject.

[0136] In some embodiments, the BCMA-associated disease or condition is a disease or condition that has relapsed in the subject to one or more previous therapies for treating the disease, and / or a disease or condition in which the subject has not responded to one or more other previous therapies for treating the disease and is therefore refractory to treatment with one or more previous therapies. In certain embodiments, the disease or condition is multiple myeloma that is relapsed or refractory disease (hereinafter also referred to as relapsed or refractory multiple myeloma or R / R multiple myeloma). In some embodiments, the subject has persistent or relapsed disease following treatment with other therapies, including, for example, another BCMA-specific antibody, and / or cells expressing a BCMA-targeted chimeric receptor, and / or chemotherapy, radiation, and / or hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some embodiments, the subject is resistant or refractory to treatment, i.e., does not respond following treatment with another BCMA-specific antibody and / or cells expressing a BCMA-targeted chimeric receptor and / or other therapy. In some embodiments, administration of T cell therapy (e.g., anti-BCMA CAR T cells) in the provided methods effectively treats the subject despite the subject having become resistant or refractory to another BCMA-targeted therapy. In some embodiments, the subject is determined to be at risk of relapse, such as not having relapsed but being at high risk of relapse, and thus the combination is administered prophylactically, e.g., to reduce the likelihood of or prevent relapse.

[0137] In some embodiments, the subject is a transplant-eligible subject, such as eligible for hematopoietic stem cell transplant (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some such embodiments, the subject has not previously undergone a transplant despite being eligible prior to administration of a BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells) and / or composition comprising the same provided herein.

[0138] In some embodiments, the subject is a transplant-ineligible subject, such as ineligible for hematopoietic stem cell transplant (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some such embodiments, such a subject is administered a BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells) and / or a composition comprising same, in accordance with the embodiments provided herein.

[0139] In some embodiments, prior to initiation of administration of the engineered cells, the subject has undergone one or more prior therapies to treat a disease or disorder, e.g., multiple myeloma. In some embodiments, the subject has undergone at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more prior therapies. In some embodiments, the subject has undergone at least 3, 4, 5, 6, 7, 8, 9, 10, or more prior therapies. In some embodiments, the subject has relapsed or is refractory to treatment with one or more prior therapies. In some embodiments, the subject has relapsed or is refractory to treatment with one to three prior therapies. In some embodiments, the subject has relapsed or is refractory to treatment with two or more prior therapies. In some embodiments, the subject has relapsed or is refractory to treatment with three or more prior therapies. In some embodiments, the subject has relapsed or is refractory to treatment with four or more prior therapies. In some embodiments, one or more prior therapies may include an immunomodulatory agent. In some embodiments, one or more prior therapies may include an anti-CD38 antibody. In some embodiments, one or more prior therapies may include a proteasome inhibitor. In some embodiments, one or more prior therapies may include an immunomodulatory agent, a proteasome inhibitor, and an anti-CD38 antibody. In some embodiments, the subject must have received at least two consecutive cycles of treatment with the prior therapy. In some embodiments, the subject must have received at least two consecutive cycles of treatment with each of the one or more prior therapies. In some embodiments, the subject must have received at least two consecutive cycles of treatment with each of the one or more prior therapies, unless the progressive disease was the best response to the regimen. In some aspects, the subject has relapsed or been refractory to one or more prior therapies. For example, the subject has R / R multiple myeloma.

[0140] In some embodiments, the methods can involve including or excluding certain subjects for therapy with BCMA-directed T cell therapy (e.g., anti-BCMA CAR T cells) or compositions comprising same based on particular criteria, diagnoses, or indicators. In some embodiments, at the time of administration of the cells or dose of conditioning lymphodepleting chemotherapy, the subject did not have active plasma cell leukemia (PCL) or a history of such. In some embodiments, if the subject had active PCL or a history of such PCL at the time of administration, the subject can be excluded from being treated according to the provided methods. In some embodiments, if the subject develops PCL, such as secondary PCL, at the time of administration, the subject can be excluded from being treated according to the provided methods. In some embodiments, evaluation of the criteria, diagnoses, or indicators can be performed at the time of screening the subject for eligibility or suitability for treatment according to the provided methods, at various steps in the treatment regimen, at the time of receiving lymphodepleting therapy, and / or at or just prior to the initiation of administration of the engineered cells or compositions thereof.

[0141] For the prevention or treatment of disease, the appropriate dosage of combination agent and / or immunotherapy, for example, T cell therapy (for example, CAR-expressing T cells) can depend on the type of disease to be treated, the specific combination agent, cell and / or recombinant receptor expressed in cell, the severity and course of disease, the route of administration, whether combination agent and / or T cell therapy is administered for prophylactic or therapeutic purposes, previous therapy, frequency of administration, the clinical history of the subject and response to cell, and the discretion of the attending physician.In some embodiments, the composition and cell are suitably administered to the subject at one time or over a series of treatments.The exemplary dosage regimen and schedule for the provided combination therapy are described.

[0142] In some embodiments, the T cell therapy and the combination agent are administered as part of a further combined treatment, which can be administered simultaneously with another therapeutic intervention or sequentially, in either order. In some contexts, the T cell therapy, e.g., engineered T cells, e.g., CAR-expressing T cells, are co-administered with another therapy sufficiently closely in time so that the T cell therapy enhances the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the cells are administered before one or more additional therapeutic agents. In some embodiments, the T cell therapy, e.g., engineered T cells, e.g., CAR-expressing T cells, is administered after one or more additional therapeutic agents. In some embodiments, the combination therapy method further comprises lymphodepleting therapy, such as administration of a chemotherapeutic agent. In some embodiments, the combination therapy further comprises administering another therapeutic agent, such as an anti-cancer agent, a checkpoint inhibitor, or another immunomodulatory agent. Uses include the use of the combination therapy in such methods and treatments, as well as the use of such compositions in the preparation of a medicament for carrying out such combination therapy methods. In some embodiments, the methods and uses thereby treat a disease or condition or disorder, such as cancer or a proliferative disease, in a subject.

[0143] Prior to, during, or following administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy) and / or combination agents, the biological activity of the T cell therapy, e.g., the biological activity of an engineered cell population, is measured in some embodiments, e.g., by any of a number of known methods. Parameters to evaluate include the ability of the engineered cells to destroy target cells, persistence of T cell activity, and other measures, such as measured using any suitable method known in the art, such as the assays described further below in Section III. In some embodiments, the biological activity of cells, e.g., T cells, administered for T cell-based therapy is measured by assaying cytotoxic cell killing, expression and / or secretion of one or more cytokines, proliferation, or expansion, such as after restimulation with an antigen. In some aspects, biological activity is measured by assessing disease burden and / or clinical outcome, such as reduction in tumor burden or burden. In some embodiments, administration of one or both agents of the combination therapy and / or repeat administration of either of the therapies can be determined based on the results of assays before, during, during, or after administration of one or both agents of the combination therapy.

[0144] In some embodiments, the combined effect of the combination agent in combination with cell therapy may be synergistic compared to treatment with the combination agent alone or cell therapy alone. For example, in some embodiments, the methods provided herein result in an increase or improvement in the desired therapeutic effect, such as an increase or improvement in the reduction or inhibition of one or more symptoms associated with cancer.

[0145] In some embodiments, the combination agent increases the expansion or proliferation of engineered T cells, such as CAR T cells. In some embodiments, the increased expansion or proliferation is observed in vivo after administration to a subject. In some embodiments, the increase in the number of engineered T cells, e.g., CAR T cells, is greater than or equal to about 1.2-fold, 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 6.0-fold, 7.0-fold, 8.0-fold, 9.0-fold, 10.0-fold, or more. Administration of AT cell therapy

[0146] In some embodiments of the methods, compositions, combinations, kits, and uses provided herein, the combination therapy comprises administering to the subject an immune cell therapy, such as a T cell therapy (e.g., a CAR-expressing T cell). In certain embodiments, the cell therapy is a BCMA-directed T cell therapy. For example, the T cell therapy is an anti-BCMA CAR T cell therapy.

[0147] In some embodiments, the cells used in or administered in connection with the provided methods contain or are engineered to contain engineered receptors, such as engineered antigen receptors, for example, chimeric antigen receptors (CARs) or T cell receptors (TCRs). Among the compositions, pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy, are particularly mentioned. Also provided are therapeutic methods for administering cells and compositions to subjects, for example, patients, in accordance with the provided methods and / or using the provided products or compositions.

[0148] In some embodiments, the cell-based therapy is or includes the administration of cells, e.g., immune cells, e.g., T cells or NK cells, that target molecules expressed on the surface of a lesion, such as a tumor or cancer. In some embodiments, the cells express a recombinant receptor, e.g., a CAR, containing an extracellular ligand-binding domain that specifically binds to the antigen. In some embodiments, the recombinant receptor is a CAR that contains an extracellular antigen-recognition domain that specifically binds to the antigen. In some embodiments, the recombinant receptor specifically binds to an antigen associated with a disease or condition, e.g., an antigen associated with or expressed on cells of a tumor or cancer. In certain embodiments, the antigen is BCMA. In some embodiments, the immune cells express a recombinant receptor, such as a transgenic TCR or chimeric antigen receptor (CAR). In some embodiments, the T cell therapy includes administering T cells engineered to express a chimeric antigen receptor (CAR). In certain embodiments, the cell therapy, e.g., anti-BCMA CAR T cell therapy, is for treating multiple myeloma, e.g., relapsed / refractory (R / R multiple myeloma). In some embodiments, the cells are autologous to the subject. In some embodiments, the cells are allogeneic to the subject. Exemplary engineered cells for administration as cell therapy in the provided methods are described in Section II.

[0149] The method for administering cells for adoptive cell therapy is known, and can be used in connection with the provided methods, compositions, and products and kits.For example, the method of adoptive T cell therapy is described, for example, in U.S. Patent Application Publication No. 2003 / 0170238 to Gruenberg et al.; U.S. Patent No. 4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85).For example, see Themeli et al. (2013) Nat Biotechnol. 31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; Davila et al. (2013) PLoS ONE 8(4):e61338.

[0150] In some embodiments, cell therapy, e.g., adoptive T cell therapy, is performed by autologous transfer, whereby cells are isolated and / or otherwise prepared from a subject to receive cell therapy or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., a patient, in need of treatment, and the cells, after isolation and processing, are administered to the same subject.

[0151] In some embodiments, cell therapy, e.g., adoptive T cell therapy, is carried out by allogeneic transfer, whereby cells are isolated and / or otherwise prepared from a subject other than the subject that is to receive or will ultimately receive cell therapy, e.g., a first subject. In such embodiments, the cells are then administered to a different subject of the same species, e.g., a second subject. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0152] The cells of the T cell therapy can be administered in a composition formulated for administration, or alternatively, in two or more compositions (e.g., two compositions) formulated for separate administration. The dose(s) of cells can include a specific or relative number of cells or engineered cells, and / or two or more subtypes in a defined ratio or composition within the composition, such as CD4 vs. CD8 T cells.

[0153] The cells can be administered by any suitable means, for example, by bolus injection, injection, e.g., intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, the cells are administered parenterally, intrapulmonary, intranasally, and, if desired for localized treatment, intralesionally. Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, a given dose is administered by a single bolus of cells. In some embodiments, a given dose is administered by multiple boluses of cells over a period of, for example, three days or less, or by continuous infusion of cells. In some embodiments, administration of the cell dose or any additional therapy, eg, lymphodepleting therapy, interventional therapy, and / or combination therapy, is performed by exogenous delivery.

[0154] For treatment of disease, the appropriate dosage will depend on the type of disease being treated, the type of cells or recombinant receptor, the severity and course of the disease, previous therapy, the subject's clinical history and response to the cells, and the discretion of the treating physician. The compositions and cells, in some embodiments, are suitably administered to the subject at one time or over a series of treatments.

[0155] In certain embodiments, the cells, or individual populations of cell subtypes, are in the range of about 1 million to about 100 billion cells, and / or that amount of cells per kilogram of body weight, e.g., 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, or a range defined by any two of the foregoing values). 0 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value therebetween, and / or any value per kilogram of body weight, are administered to a subject. Dosages may vary depending on the specific characteristics of the disease or disorder and / or the patient and / or other treatments.

[0156] In some embodiments, for example, when the subject is a human, the dose is about 1 x 10 8 Fewer than 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs), e.g., about 1 x 10 6 ~1×10 8 In the range of 2 x 10 such cells, for example 6 , 5×10 6 , 1×10 7 , 5×10 7 Or 1 x 10 8, or a range between any two of the foregoing values. Exemplary dosages for use or administration in accordance with the provided methods are provided in Section IA-2 below.

[0157] The cells can be administered by any suitable means. The cells are administered in a dosage regimen to achieve a therapeutic effect, such as reducing tumor burden. Dosage and administration can depend in part on the schedule of administration of the combination agent, which can be administered before, after, and / or simultaneously with the start of administration of T cell therapy. Various dosage schedules for T cell therapy include, but are not limited to, single or multiple doses over various time points, bolus administration, and pulse infusion.

[0158] Preconditioning a subject with immunodepleting (eg, lymphodepleting) therapy can, in some aspects, improve the efficacy of adoptive cellular therapy (ACT).

[0159] Thus, in some embodiments, the method includes administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or a combination thereof, to the subject prior to the initiation of cell therapy. For example, the subject can be administered the preconditioning agent at least 2 days, e.g., at least 3, 4, 5, 6, or 7 days, prior to the initiation of cell therapy. In some embodiments, the subject is administered the preconditioning agent 7 days or less, e.g., 6, 5, 4, 3, or 2 days or less, prior to the initiation of cell therapy.

[0160] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose of 20 mg / kg to 100 mg / kg or an approximation thereof, e.g., 40 mg / kg to 80 mg / kg or an approximation thereof. In some aspects, the subject is preconditioned with cyclophosphamide at or an approximation thereof. In some embodiments, the cyclophosphamide can be administered in a single dose or in multiple doses, e.g., daily, every other day, or every third day. In some embodiments, the cyclophosphamide is administered once daily for one or two days. In some embodiments, when the lymphodepleting agent includes cyclophosphamide, the subject is preconditioned with cyclophosphamide at a dose of 100 mg / m 2 ~500mg / m 2 or between or between these approximate numbers, e.g., 200 mg / m 2 ~400mg / m 2 or 250 mg / m 2 ~350mg / m 2 In some instances, the subject is administered a dose of cyclophosphamide of about 300 mg / m 2 In some embodiments, the cyclophosphamide is administered daily for, for example, 1-5 days, e.g., 3-5 days, etc. In some instances, the subject receives about 300 mg / m 2 of cyclophosphamide will be administered daily for three days prior to the start of cell therapy.

[0161] In some embodiments, when the lymphodepleting agent comprises fludarabine, the subject receives 1 mg / m 2 ~100mg / m 2 or between or between these approximate numbers, e.g., 10 mg / m 2 ~75mg / m 2 , 15 mg / m 2 ~50mg / m 2 , 20 mg / m 2~40mg / m 2 or 24 mg / m 2 ~35mg / m 2 In some instances, the subject is administered a dose of fludarabine of about 30 mg / m 2 In some embodiments, the subject receives about 30 mg / m of fludarabine. In some embodiments, the fludarabine can be administered in a single dose or in multiple doses, for example, given daily, every other day, or every third day. In some embodiments, the fludarabine is administered daily for 1-5 days, such as for 3-5 days. In some instances, the subject receives about 30 mg / m 2 of fludarabine will be administered daily for three days prior to the start of cell therapy.

[0162] In some embodiments, the lymphodepleting agent comprises a combination of drugs, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of drugs can include cyclophosphamide at any dose or administration schedule, such as those described above, and fludarabine at any dose or administration schedule, such as those described above. For example, in some aspects, the subject receives cyclophosphamide at a dose of 60 mg / kg (approximately 2 g / m 2 ) of cyclophosphamide and 3 to 5 doses of 25 mg / m 2 Fludarabine is administered before the first or subsequent dose.

[0163] After administration of the cells, the biological activity of the engineered cell population is measured, in some embodiments, by any of a number of known methods, for example. Parameters for evaluation include specific binding of engineered or natural T cells or other immune cells to antigens in vivo, for example, by imaging, or ex vivo, for example, by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable known method, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells is measured by assaying the expression and / or secretion of one or more cytokines, such as CD107a, IFNγ, IL-2, and TNF. In some aspects, biological activity is measured by assessing clinical outcomes, such as reduction in tumor burden or burden. 1. Compositions and Formulations

[0164] In some embodiments, the dose of cells of a T cell therapy, such as a T cell therapy comprising cells engineered with a recombinant antigen receptor, e.g., a CAR or TCR, is provided as a composition or formulation, such as a pharmaceutical composition or formulation. Such compositions can be used in accordance with the provided methods, such as in the prevention or treatment of diseases, conditions, and disorders, such as in the treatment of multiple myeloma, e.g., relapsed or refractory multiple myeloma.

[0165] In some embodiments, T cell therapies, such as engineered T cells (e.g., CAR T cells), are formulated with a pharmaceutically acceptable carrier. In some aspects, the choice of carrier is determined, in part, by the particular cell or agent and / or by the method of administration. Accordingly, a variety of suitable formulations exist. For example, the pharmaceutical composition can contain a preservative. Suitable preservatives can include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some aspects, a mixture of two or more preservatives is used. The preservative or mixtures thereof are typically present in an amount of about 0.0001% to about 2% by weight of the total composition. Carriers are described, for example, in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed and may include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polymers; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as polyethylene glycol (PEG).

[0166] In some embodiments, a buffering agent is included in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some embodiments, a mixture of two or more buffering agents is used. The buffering agent or mixture thereof is typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0167] The formulation may comprise an aqueous solution. The formulation or composition may contain two or more active ingredients useful for the particular indication, disease, or condition being prevented or treated by the cells or agent, where the activities of each do not adversely affect each other. Such active ingredients are suitably present in combination in amounts effective for the intended purpose. Thus, in some embodiments, the pharmaceutical composition further comprises another pharmaceutically active agent or drug, such as a chemotherapeutic agent, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0168] In some embodiments, the pharmaceutical composition contains cells in an amount effective for treating or preventing a disease or condition, such as a therapeutically or prophylactically effective amount. In some embodiments, therapeutic or prophylactic effectiveness is monitored by periodic evaluation of the treated subject. For repeated administration over several days or longer, depending on the condition, treatment is repeated until the desired suppression of disease symptoms occurs. However, other dosage regimens may be useful and can be determined. The desired dosage can be delivered by a single bolus administration of the composition, by multiple bolus administration of the composition, or by continuous infusion administration of the composition.

[0169] Cells can be administered using standard administration techniques, formulations, and / or devices. Formulations and devices, such as syringes and vials, are provided for storing and administering the compositions. With respect to cells, administration can be autologous or xenogeneic. For example, immunoresponsive cells or precursors can be obtained from a subject and administered to the same subject or a different compatible subject. Peripheral blood-derived immunoresponsive cells or their progeny (e.g., derived in vivo, ex vivo, or in vitro) can be administered by localized injection, including catheter administration, systemic injection, localized injection, intravenous injection, or parenteral administration. When administering a therapeutic composition (e.g., a pharmaceutical composition containing genetically modified immunoresponsive cells), it will generally be formulated in a unit-dosage injectable form (solution, suspension, emulsion).

[0170] Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the agent or cell population is administered parenterally. The term "parenteral," as used herein, includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. In some embodiments, the agent or cell population is administered to a subject using peripheral systemic delivery via intravenous, intraperitoneal, or subcutaneous injection.

[0171] In some embodiments, the composition is provided as a sterile liquid preparation, such as an isotonic aqueous solution, suspension, emulsion, dispersion, or viscous composition, which may be buffered to a selected pH in some aspects. Liquid preparations are usually easier to prepare than gels, other viscous compositions, and solid compositions. Furthermore, liquid compositions are somewhat easier to administer, especially by injection. On the other hand, viscous compositions can be formulated within an appropriate viscosity range to provide a longer contact period with specific tissues. Liquid or viscous compositions can include, for example, a solvent or dispersion medium carrier, including water, saline, phosphate buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), and suitable mixtures thereof.

[0172] Sterile injectable solutions can be prepared by incorporating the cells in a solvent such as a mixture with a suitable carrier, diluent, or excipient, such as sterile water, saline, glucose, dextrose, or the like. The composition may also be lyophilized. The composition may contain auxiliary substances such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity-enhancing additives, preservatives, flavoring agents, coloring agents, and the like, depending on the desired route of administration and preparation. Standard texts can, in some embodiments, be consulted for preparing suitable preparations.

[0173] Various additives can be added to enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of microbial action can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0174] Formulations to be used for in vivo administration are generally sterile. Sterility is readily accomplished, for example, by filtration through sterile filtration membranes.

[0175] For the prevention or treatment of disease, the appropriate dosage may depend on the type of disease being treated, the type of agent(s), the type of cells or recombinant receptor, the severity and course of the disease, whether the agent or cells are being administered for prophylactic or therapeutic purposes, previous therapy, the subject's clinical history and response to the agent or cells, and the discretion of the attending physician. The compositions, in some embodiments, are suitably administered to the subject at one time or over a series of treatments.

[0176] In some cases, cell therapy is administered as a single pharmaceutical composition containing cells. In some embodiments, a given dose is administered by a single bolus of cells or drug. In some embodiments, it is administered by multiple bolus administration of cells or drug over a period of, for example, 3 days or less, or by continuous infusion of cells or drug. 2. Dosage Schedule and Administration

[0177] In some embodiments, the dose of cells is administered to the subject in accordance with the provided combination therapy method.In some embodiments, the size or timing of the dose is determined as a function of the specific disease or condition in the subject.In consideration of the provided explanation, the size or timing of the dose for a specific disease can be empirically determined.

[0178] In certain embodiments, the cells, or individual populations or subtypes of cells, are present in a range of about 100,000 to about 100 billion cells, and / or that amount of cells per kilogram of subject body weight, e.g., 100,000 to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells. cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells The subject is administered between about 120 million cells and about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value between these ranges, and / or per kilogram of the subject's body weight. Dosages may vary depending on the specific characteristics of the disease or disorder and / or patient and / or other treatment. In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells; in other embodiments, it refers to the number of T cells or PBMCs or total cells administered.

[0179] In some embodiments, the cell therapy comprises at least or at least about 0.1 x 10 6 Cells / kg (subject weight), 0.2 × 10 6 cells / kg, 0.3×10 6 cells / kg, 0.4×106 cells / kg, 0.5×10 6 cells / kg, 1×10 6 cells / kg, 2.0×10 6 cells / kg, 3×10 6 cells / kg or 5 x 10 6 It includes administering a dose containing a number of cells that is or is about cells / kg.

[0180] In some embodiments, the cell therapy comprises 0.1 x 10 6 Cells / kg (subject weight) ~ 1.0 x 10 7 Between or around 0.5 x 10 cells / kg 6 cells / kg~5×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~3×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~2×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~1×10 6 Between or about 1.0 x 10 cells / kg 6 ~5 x 10 cells / kg (subject weight) 6 Between or about 1.0 x 10 cells / kg 6 cells / kg~3×10 6 Between or about 1.0 x 10 cells / kg 6 cells / kg~2×10 6 Between or about 2.0 x 10 cells / kg 6 ~5 x 10 cells / kg (subject weight) 6 Between or about 2.0 x 10 cells / kg 6 cells / kg~3×10 6 cells / kg or 3.0 x 10 6 ~5 x 10 cells / kg (subject weight) 6 This includes administering a dose containing a number of cells between or about (inclusive) cells / kg.

[0181] In some embodiments, the dose of cells is 2×10 5 or approximate number of cells / kg ~ 2 x 10 6 or approximately 4 x 10 cells / kg 5 or approximately 1 x 10 cells / kg 6 Between 6 x 10 or the approximate number of cells / kg 5 or approximately 8 x 10 cells / kg 5 In some embodiments, the dose of cells is between 2 x 10 cells / kg or about 2 x 10 cells / kg of subject body weight. 5 3×10 or less cells (e.g., antigen-expressing, e.g., CAR-expressing cells) (cells / kg), e.g., 3×10 5 Cells / kg or less, 4 x 10 5 Cells / kg or less, 5 x 10 5 Cells / kg or less, 6 x 10 5 7 x 10 cells / kg or less 5 Cells / kg or less, 8 x 10 5 Cells / kg or less, 9 x 10 5 Cells / kg or less, 1 x 10 6 Cells / kg or less, or 2 x 10 6 In some embodiments, the dose of cells comprises at least about 2 x 10 cells / kg or less. 5 or at or about that number of cells (e.g., antigen-expressing, e.g., CAR-expressing cells) (cells / kg), e.g., at least or at least about 3 x 10 5 cells / kg or an approximation thereof, at least or at least about 4 x 10 5 cells / kg or an approximation thereof, at least or at least about 5 x 10 5 cells / kg or an approximation thereof, at least or at least about 6 x 10 5 cells / kg or an approximation thereof, at least or at least about 7 x 10 5cells / kg or an approximation thereof, at least or at least about 8 x 10 5 cells / kg or an approximation thereof, at least or at least about 9 x 10 5 cells / kg or an approximation thereof, at least or at least about 1 x 10 6 cells / kg or at or about that number, or at least about 2 x 10 6 Includes cells / kg or any value or approximation thereof.

[0182] In some embodiments, the dose of cells is a flat dose of cells or a fixed dose of cells such that the dose of cells is not related to or based on the body surface area or weight of the subject.

[0183] In some embodiments, the cell therapy comprises 1 x 10 5 ~2×10 9 5 x 10 or approximately 5 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 5 ~1×10 9 or approximately 1 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), or 1 x 10 6 ~1×10 9 In some embodiments, the cell therapy comprises administering a dose comprising at or about 1 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), inclusive. 5 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), for example, at least or about at least 1 x 10 6 , at least or about at least 1 x 10 7 , at least or about at least 1 x 10 8 , at least or about at least 1 x 10 9 It includes administering a dose of cells, including a number of such cells.

[0184] In some embodiments, the dose of genetically engineered cells is at least or at least about 1 x 10 5 CAR-expressing cells, at least or at least about 2.5 x 10 5 CAR-expressing cells, at least or at least about 5 x 10 5 CAR-expressing cells, at least or at least about 1 x 10 6 CAR-expressing cells, at least or at least about 2.5 x 10 6 CAR-expressing cells, at least or at least about 5 x 10 6 CAR-expressing cells, at least or at least about 1 x 10 7 CAR-expressing cells, at least or at least about 2.5 x 10 7 CAR-expressing cells, at least or at least about 5 x 10 7 CAR-expressing cells, at least or at least about 1 x 10 8 CAR-expressing cells, at least or at least about 2.5 x 10 8 CAR-expressing cells, or at least about or at least 5 x 10 8 Contains CAR-expressing cells.

[0185] In some embodiments, for example, when the subject is a human, the dose is 1×10 6 or greater than approximately 2 x 10 total recombinant receptor (e.g., CAR)-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) 9 or less than the approximate number of total recombinant receptor (e.g., CAR)-expressing cells, T cells or peripheral blood mononuclear cells (PBMCs), e.g., 1.0 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 1.0 x 10 such cells, e.g., 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×108 , 8×10 8 or 1.2 x 10 9 In some embodiments, for example, when the subject is a human, the dose is 1 x 10 or about 1 x 10 such total cells. 6 or greater than approximately 2 x 10 total recombinant receptor (e.g., CAR)-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) 9 or less than the approximate number of total recombinant receptor (e.g., CAR)-expressing cells, T cells or peripheral blood mononuclear cells (PBMCs), e.g., 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 2.5 x 10 such cells 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, for example, when the subject is a human, the dose is 1.0 x 10 or about 1.0 x 10 such total cells. 7 In some embodiments, for example, when the subject is a human, the dose comprises 1.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 2.0 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 2.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 5×10 or approximately 5×10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7In some embodiments, for example, when the subject is a human, the dose comprises 1.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 3 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 4.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 6 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, if the subject is a human, the dose comprises 8 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 1.2 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 9 The cells comprise at or about the total number of recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0186] In some embodiments, the dose of genetically engineered cells is 1×10 5 pieces or the approximate number ~ 2 x 10 9 Total CAR-expressing (CAR+) T cells, 1 x 10 5 Pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5Pieces or the approximate number ~ 5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 Pieces or the approximate number ~ 5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 6 Pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 Pieces or the approximate number ~ 5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 Pieces or the approximate number ~ 5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 6 Total CAR-expressing T cells, 2.5 x 10 6 Pieces or the approximate number ~ 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 2.5 x 10 8Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 Pieces or the approximate number ~ 5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 1 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 Pieces or the approximate number ~ 5 x 10 6 5 x 10 total CAR-expressing T cells or their approximate number 6 Pieces or the approximate number ~ 5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 Pieces or the approximate number ~ 5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 7 Pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 Pieces or the approximate number ~ 5 x 10 71 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 7 Pieces or the approximate number ~ 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 pieces or its approximate number ~ 2.5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 pieces or its approximate number ~ 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 Pieces or the approximate number ~ 5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 7 Pieces or the approximate number ~ 5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 8 Pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 8 pieces or its approximate number ~ 2.5 x 10 8 or approximately 2.5 x 10 total CAR-expressing T cells 8 Pieces or the approximate number ~ 5 x 10 8 In some embodiments, the dose of engineered cells comprises at or about 1.0 x 10 total CAR-expressing T cells. 7 pieces or the approximate number ~ 8 x 10 8 Total CAR-expressing (CAR+) T cells, 1.0 x 10 7 pieces or its approximate number ~ 6.5 x 10 8 Total CAR+ T cells, 1.5 x 10 7 pieces or its approximate number ~ 6.5 x 10 8 Total CAR+ T cells, 1.5 x 10 7pieces or its approximate number ~ 6.0 x 10 8 Total CAR+ T cells, 2.5 x 10 7 pieces or its approximate number ~ 6.0 x 10 8 or approximately 5.0 x 10 total CAR+ T cells 7 pieces or its approximate number ~ 6.0 x 10 8 Contains at or about the total number of CAR+ T cells.

[0187] In some embodiments, the dose of genetically engineered cells is 2.5×10 7 Approximately 1.2 x 10 or approximately 1.2 x 10 CAR-expressing (CAR+) T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 9 Between 5.0 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total PBMCs 7 or approximate number of CAR-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) ~6.0 x 10 8 Between 5.0 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total PBMCs 7 or approximate number of CAR-expressing T cells ~4.5 x 10 8 Between 1.5 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 8 or approximate number of CAR-expressing T cells ~3.0 x 10 8 In some embodiments, the dose comprises between 2.5 x 10 and approximately 2.5 x 10 CAR-expressing T cells, total T cells, or total PBMCs, each inclusive. In some embodiments, the number refers to the total number of CD3+ or CD8+, and in some cases, CAR-expressing (e.g., CAR+) cells. In some embodiments, the dose is between 2.5 x 10 and approximately 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 5.0 x 10 or the approximate number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 Pieces or their approximate number ~ 6.0 x 10 8 5.0 x 10 or the approximate number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 Pieces or their approximate number ~ 4.5 x 10 8or approximately 1.5 x 10 CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 8 Pieces or their approximate number ~ 3.0 x 10 8 The number of cells includes the number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells (inclusive) at or near that number.

[0188] In some embodiments, the dose of engineered cells refers to the total number of CD3+ CAR-expressing (CAR+) or CD4+ / CD8+ CAR-expressing (CAR+) cells. In some embodiments, the dose is 1.0 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 1.5 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 1.2 x 10 9 2.0 x 10 or approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 1.2 x 10 9 2.5 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 1.2 x 10 9 5.0 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 6.0 x 10 8 5.0 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 4.5 x 10 8 or approximately 1.5 x 10 CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 8 Pieces or their approximate number ~ 3.0 x 108 In some embodiments, the dose comprises 1.0 x 10 or approximately 1.0 x 10 total CD3+ or CD4+ / CD8+ T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells (inclusive) of engineered cells. 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, the dose comprises 2.5 x 10 total CD3+ or CD4+ / CD8+ T cells or CD3+ CAR-expressing or CD4+ / CD8+ CAR-expressing cells. 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, the dose comprises 1.0 x 10 CD3+ CAR-expressing cells or approximately 1.0 x 10 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 The cells contain at or about that number of CD4+ / CD8+ CAR-expressing cells.

[0189] In some embodiments, the dose is 1.0 x 10 7 In some embodiments, the dose is 1.5 x 10 7In some embodiments, the dose is 2.0 x 10 7 In some embodiments, the dose is 2.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 2.5 x 10 7 In some embodiments, the dose is 5×10 7 In some embodiments, the dose is 1.5 x 10 8 In some embodiments, the dose is 3 x 10 8 In some embodiments, the dose is 4.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 4.5 x 10 8 In some embodiments, the dose is 6 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 6 x 10 8 In some embodiments, the dose is 8 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 8 x 10 8 In some embodiments, the dose is 1.2 x 10 9 In some embodiments, the dose is 2.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 2.5 x 10 7 In some embodiments, the dose is 5 x 10 7 In some embodiments, the dose is 1.5 x 10 8 In some embodiments, the dose is 3 x 10 8 In some embodiments, the dose is 4.5 x 10 8 In some embodiments, the dose is 6 x 10 8 In some embodiments, the dose is 6.5 x 10 8In some embodiments, the dose is 8 x 10 8 In some embodiments, the dose is 1.2 x 10 9 or approximately that number of CD4+ or CD8+ CAR-expressing cells.

[0190] In some embodiments, the dose of T cells comprises CD4+ T cells, CD8+ T cells, or CD4+ and CD8+ T cells.

[0191] In some embodiments, for example, when the subject is a human, the dose of CD4+ T cells and CD8+ T cells totals 1×10 6 Pieces or their approximate number ~ 2 x 10 9 Between or about 2.5 x 10 total CAR-expressing CD4+ cells and CAR-expressing CD8+ cells, for example, 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 5 x 10 such cells 7 Pieces or their approximate number ~ 4.5 x 10 8 in the range of or about that number of such cells; e.g., 1.0 x 10 7 Pieces or its approximate number, 2.5 x 10 7 Pieces or its approximate number, 2.0 x 10 7 Pieces or its approximate number, 2.5 x 10 7 Pieces or its approximate number, 5 x 10 7 Pieces or its approximate number, 1.5 x 10 8 Pieces or its approximate number, 3 x 10 8 Pieces or its approximate number, 4.5 x 10 8 Pieces or its approximate number, 6 x 10 8 pieces or its approximate number, 6.5 x 10 8 Pieces or its approximate number, 8 x 10 8 or its approximate number, or 1.2 x 10 9In some embodiments, for example, when the subject is a human, the dose of CD8+ T cells, including in the dose comprising CD4+ T cells and CD8+ T cells, is 1 x 10 or about 1 x 10 such total cells, or a range between any two of the foregoing values. 6 Pieces or their approximate number ~ 2 x 10 9 Between or about 10 total recombinant receptor (e.g., CAR)-expressing CD8+ cells, e.g., 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 5 x 10 such cells 7 Pieces or their approximate number ~ 4.5 x 10 8 in the range of or about that number of such cells; e.g., 2.5 x 10 7 Pieces or its approximate number, 5 x 10 7 Pieces or its approximate number, 1.5 x 10 8 Pieces or its approximate number, 3 x 10 8 Pieces or its approximate number, 4.5 x 10 8 Pieces or its approximate number, 6 x 10 8 Pieces or its approximate number, 8 x 10 8 or its approximate number, or 1.2 x 10 9 This includes at or about that number of such total cells, or a range between any two of the foregoing values.

[0192] In some embodiments, a dose of cells, e.g., recombinant receptor-expressing T cells, is administered to a subject as a single dose, or is administered only once within a period of two weeks, one month, three months, six months, one year, or more. In some embodiments, a patient is administered multiple doses, and each or the total dose can be within any of the values ​​set forth above. In some embodiments, the engineered cells or compositions of engineered cells for administration exhibit characteristics indicative of or consistent with cell health. In some embodiments, 70, 75, 80, 85, or 90% of such doses, or at least about 70, 75, 80, 85, or 90% of the CAR+ cells exhibit one or more characteristics or phenotypes indicative of cell health or biologically active CAR cells, such as the absence of expression of apoptotic markers.

[0193] In certain embodiments, the phenotype is or includes the absence of apoptosis and / or the absence of indicators that the cell is undergoing the apoptotic process. Apoptosis is a process of programmed cell death that involves a series of stereotypical morphological and biochemical events that result in characteristic cellular changes and death, including blebbing, cell shrinkage, nuclear fragmentation, chromatin condensation, chromosomal DNA fragmentation, and general mRNA decay. In some aspects, the early stage of apoptosis can be indicated by the activation of certain caspases, such as 2, 8, 9, and 10. In some aspects, the mid- to late-stage of apoptosis is characterized by further loss of membrane integrity, chromatin condensation, and DNA fragmentation, and includes biochemical events such as the activation of caspases 3, 6, and 7.

[0194] In certain embodiments, the phenotype is one or more factors associated with programmed cell death, e.g., pro-apoptotic factors known to initiate apoptosis, e.g., members of the death receptor pathway, activated members of the mitochondrial (intrinsic) pathway, e.g., Bcl-2 family members, e.g., Bax, Bad, and Bid, and negative expression of caspases. In certain embodiments, the phenotype is an indicator, e.g., the absence of annexin V molecules or TUNEL staining, which will preferentially bind to cells undergoing apoptosis when incubated or contacted with the cell composition. In some embodiments, the phenotype is or includes expression of one or more markers indicative of an apoptotic state in cells. In some embodiments, the phenotype is the lack of expression and / or activation of a caspase, e.g., caspase-3. In some aspects, activation of caspase-3 indicates increased or resurrected apoptosis. In certain embodiments, caspase activation can be detected by known methods. In some embodiments, caspase activation can be detected using an antibody that specifically binds to activated caspase (i.e., specifically binds to the cleaved polypeptide). In certain embodiments, the phenotype is or comprises active caspase 3. In some embodiments, the marker of apoptosis is a reagent that detects a cellular feature associated with apoptosis. In certain embodiments, the reagent is an annexin V molecule.

[0195] In some embodiments, compositions containing engineered cells for administration contain a certain number or amount of cells that exhibit a phenotype indicative of or consistent with cellular health. In some of any of the embodiments, less than about 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the CAR-expressing T cells in a dose of engineered T cells express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any of the embodiments, less than 5%, 4%, 3%, 2%, or 1% of the CAR-expressing T cells in a dose of engineered T cells express Annexin V or active Caspase 3.

[0196] In the context of adoptive cell therapy, administration of a given "dose" of cells encompasses administration of a given amount or number of cells as a single composition and / or a single uninterrupted administration, e.g., as a single injection or continuous infusion, as well as administration of a given amount or number of cells as divided doses provided in multiple individual compositions or infusions over a specified period of time, which is three days or less. Thus, in some situations, a dose is a single or continuous administration of a specified number of cells given or initiated at a single time point. However, in some situations, a dose is administered in multiple injections or infusions over a period of three days or less, such as once daily for three or two days, or by multiple infusions over a one-day period.

[0197] Thus, in some aspects, the dose of cells is administered in a single pharmaceutical composition, hi some embodiments, the dose of cells is administered in multiple compositions that collectively contain the dose of cells.

[0198] The term "split dose" refers to a dose that is divided so as to be administered over more than one day. This type of administration is encompassed by the present method and is considered to be a single dose. In some embodiments, the split dose cells are administered in multiple compositions that collectively comprise the dose cells over a period of 3 days or less.

[0199] Thus, the dose of cells can be administered as a split dose. For example, in some embodiments, the dose can be administered to a subject over two or three days. An exemplary method for split dosing includes administering 25% of the dose on day 1 and the remaining 75% of the dose on day 2. In other embodiments, 33% of the dose can be administered on day 1 and the remaining 67% on day 2. In some aspects, 10% of the dose is administered on day 1, 30% of the dose is administered on day 2, and 60% of the dose is administered on day 3. In some embodiments, the split doses are not spread out over more than three days.

[0200] In some embodiments, the dose of cells is generally large enough to be effective in reducing disease burden.

[0201] In some embodiments, cells are administered at a desired dosage, which in some aspects includes a desired dose or number of cells or cell type(s) and / or a desired ratio of cell types. Thus, the dosage of cells is, in some embodiments, based on the total number of cells (or number per kg body weight) and the desired ratio of individual populations or subtypes, e.g., the ratio of CD4+ to CD8+. In some embodiments, the dosage of cells is based on the desired total number (or number per kg body weight) of cells in each population or of each cell type. In some embodiments, the dosage is based on a combination of such features, such as the desired number of total cells, the desired ratio, and the desired total number of cells in each population.

[0202] In some embodiments, CD8 + and CD4 +A population or subtype of cells, such as T cells, is administered at a desired dose of total cells, such as a desired dose of T cells, or within an acceptable difference. In some embodiments, the desired dose is a desired number of cells, or a desired number of cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some embodiments, the desired dose is at or exceeds a minimum number of cells or a minimum number of cells per unit of body weight. In some embodiments, among the total cells administered at a desired dose, individual populations or subtypes are administered at a desired output ratio (e.g., CD4 + Against CD8 + ) within a certain allowable difference or error in such ratio.

[0203] In some embodiments, cells are administered at a desired dose of one or more individual populations or subtypes of cells, such as a desired dose of CD4+ cells and / or a desired dose of CD8+ cells, or within an acceptable difference. In some aspects, the desired dose is the desired number of cells of a subtype or population, or the desired number of such cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is at or exceeds the minimum number of cells of a population or subtype, or the minimum number of cells of a population or subtype per unit of body weight.

[0204] Thus, in some embodiments, dosage is based on a desired fixed dose and desired ratio of total cells and / or based on a desired fixed dose of one or more, e.g., each, of individual subtypes or subpopulations. Thus, in some embodiments, dosage is based on a desired fixed or minimum dose of T cells and CD4 + Against CD8 + Based on the desired ratio of cells and / or CD4 + and / or CD8 + Based on desired fixation or minimum dose of cells.

[0205] In some embodiments, cells are administered at a desired output ratio or within an acceptable range of multiple cell populations or subtypes, such as CD4+ and CD8+ cells or subtypes. In some aspects, the desired ratio can be a specific ratio or a range of ratios. For example, in some embodiments, the desired ratio (e.g., CD4 + Against CD8 + The ratio of cells is between 5:1 or about to 5:1 or about to (or greater than about 1:5 and less than about 5:1), or between 1:3 or about to 3:1 or about to (or greater than about 1:3 and less than about 3:1), for example, between 2:1 or about to 1:5 or about to (or greater than about 1:5 and less than about 2:1), for example, 5:1, 4.5:1, 4:1, 3.5:1, 3:1 , 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5 or an approximation thereof. In some embodiments, acceptable differences are within about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the desired ratio, including any value in between these ranges.

[0206] In some embodiments, the cell dose or composition comprises a defined or target ratio of CD4+ cells expressing the recombinant receptor to CD8+ cells expressing the recombinant receptor and / or CD4+ cells to CD8+ cells that is approximately 1:1 or between approximately 1:3 and approximately 3:1, e.g., approximately 1:1.

[0207] In certain embodiments, the number and / or concentration of cells refers to the number of recombinant receptor (e.g., CAR)-expressing cells. In other embodiments, the number and / or concentration of cells refers to the number or concentration of administered total cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0208] In some embodiments, the size of the dose is determined based on one or more criteria such as the subject's response to previous treatment, e.g., chemotherapy; the disease burden in the subject, e.g., tumor burden, volume, size or extent, degree or type of metastasis, stage; and / or the likelihood or incidence that the subject will develop a toxic outcome, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or host immune response, to the cells and / or recombinant receptor being administered.

[0209] In some embodiments, administration of a combination agent in combination with cells can significantly increase the expansion or proliferation of the cells, thereby allowing a lower dose of cells to be administered to the subject. In some cases, the provided methods allow for a lower dose of such cells to be administered, such as 1.5, 2, 3, 4, 5, or 10 times lower than the dose in methods in which cell therapy is administered without administration of a combination agent, to achieve the same or better treatment efficacy as the dose in methods in which cell therapy is administered without administration of a combination agent.

[0210] In some embodiments, for example, the dose is 5.0 x 10 6 ~2.25×10 7 , 5.0×10 6 ~2.0×10 7 , 5.0×10 6 ~1.5×10 7 , 5.0×10 6 ~1.0×10 7 , 5.0×10 6 ~7.5×10 6 , 7.5×10 6 ~2.25×10 7 , 7.5×10 6 ~2.0×10 7 , 7.5×10 6 ~1.5×10 7 , 7.5×10 6 ~1.0×10 7 , 1.0×10 7 ~2.25×10 7 , 1.0×10 7 ~2.0×107 , 1.0×10 7 ~1.5×10 7 , 1.5×10 7 ~2.25×10 7 , 1.5×10 7 ~2.0×10 7 , 2.0×10 7 ~2.25×10 7 In some embodiments, the dose of cells comprises at least or at least about 5 x 10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 10×10 6 ~Approx. 15×10 6 In some embodiments, such a dose, e.g., such a target number of cells, refers to the total recombinant receptor-expressing cells in the administered composition.

[0211] In some embodiments, for example, the lower dose is about 5×10 per kilogram of subject body weight. 6 Fewer than about 4.5 x 10 cells, recombinant receptor (e.g., CAR)-expressing cells, T cells and / or PBMCs, e.g., about 4.5 x 10 per kilogram of subject body weight 6 , 4×10 6 , 3.5×10 6 , 3×10 6 , 2.5×10 6 , 2 × 10 6 , 1.5×10 6 , 1×10 6 , 5×10 5 , 2.5×10 5 or 1×10 5 In some embodiments, the lower dose contains less than about 1 x 10 such cells per kilogram of subject body weight. 5 , 2 × 10 5 , 5×10 5 or 1×10 6In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells; in other embodiments, they refer to the number of T cells or PBMCs or total cells administered.

[0212] In some embodiments, the subject receives multiple doses of cells, for example, two or more doses or multiple sequential doses. In some embodiments, two doses are administered to the subject. In some embodiments, the subject receives sequential doses, for example, the second dose is administered approximately 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days after the first dose. In some embodiments, the first dose is followed by multiple sequential doses, such that the additional dose(s) are administered following the administration of the sequential doses. In some aspects, the number of cells administered to the subject in the additional doses is the same or similar to the first dose and / or the sequential doses. In some embodiments, the additional dose(s) is / are greater than the previous dose. In some embodiments, one or more subsequent doses of cells can be administered to the subject. In some embodiments, the subsequent dose of cells is administered more than or about 7, 14, 21, 28, or 35 days after the start of administration of the first dose of cells. The subsequent dose of cells may be more than, about the same as, or less than the first dose. In some embodiments, the administration of T cell therapy may be repeated, such as administering the first and / or second dose of cells.

[0213] In some embodiments, the administration of the cell therapy, e.g., the start of the dose of cells or the first dose of a divided dose of cells, is administered prior to (before), simultaneously with, or after (subsequently or subsequent to) the administration of the combination agent.

[0214] In some embodiments, the dose of cells or the subsequent dose of cells is administered simultaneously with the start of administration of the combination agent in the combination therapy method. In some embodiments, the dose of cells or the subsequent dose of cells is administered on the same day as the start of administration of the combination agent in the combination therapy method. In some embodiments, the dose of cells or the subsequent dose of cells is administered within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days of the start of administration of the combination agent in the combination therapy method.

[0215] In some embodiments, the dose of cells or subsequent doses of cells are administered before beginning or commencing administration of a combination agent in accordance with a provided combination therapy. In some embodiments, the dose of cells is administered at least or at least about 1 hour, at least or at least about 2 hours, at least or at least about 3 hours, at least or at least about 6 hours, at least or at least about 12 hours, at least or at least about 1 day, at least or at least about 2 days, at least or at least about 3 days, at least or at least about 4 days, at least or at least about 5 days, at least or at least about 6 days, at least or at least about 7 days, at least or at least about 12 days, at least or at least about 14 days, at least or at least about 15 days, at least or at least about 21 days, at least or at least about 28 days, at least or at least about 30 days, at least or at least about 35 days, at least or at least about 42 days, at least or at least about 60 days, or at least or at least about 90 days before administration of a combination agent in accordance with a provided combination therapy.

[0216] In some embodiments, the administration of a combination agent in accordance with a provided combination therapy occurs at a time when the prior administration of immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy) is associated or may be associated with decreased functionality of T cells compared to functionality of T cells at a time immediately prior to initiation of immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy) or at an earlier time point after initiation of T cell therapy. In some embodiments, the method involves evaluating a sample from the subject following administration of a dose of cells of T cell therapy, e.g., adoptive T cell therapy, but prior to administration of a combination agent, for one or more functions of T cells, such as expansion or persistence of cells as determined by, e.g., blood levels or amounts, or other phenotypes or desired outcomes described herein, e.g., those described in Section III. In some embodiments, the method involves evaluating a sample from the subject following administration of a dose of cells of T cell therapy, e.g., adoptive T cell therapy, but prior to administration of a combination agent, for expression of one or more exhaustion markers. Various parameters for determining or evaluating a combination therapy regimen are described in Section III. B. Administration of Combination Drugs

[0217] In some embodiments, the provided combination therapy methods, compositions, combinations, kits, and uses involve the administration of a combination agent, which can be administered before, after, during, simultaneously with, or near-simultaneously with the administration of T cell therapy, e.g., T cells expressing a chimeric antigen receptor (CAR). In some embodiments, the combination agent includes an immunomodulatory compound, e.g., a structural or functional analog or derivative of thalidomide and / or an inhibitor of E3 ubiquitin ligase, e.g., Compound A (Iberdomide, (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione). In some embodiments, the combination agent includes an immunomodulatory agent, e.g., Compound A, and a steroid, e.g., dexamethasone. In some embodiments, the combination includes a standard triplet regimen, e.g., a drug given as part of a standard cancer or proliferative disease treatment, e.g., MM treatment. In some embodiments, the combination includes daratumumab, pomalidomide, and dexamethasone, and the standard triplet regimen is the DPd regimen. In some embodiments, the combination includes pomalidomide, bortezomib, and dexamethasone, and the standard triplet regimen is the PVd regimen. 1. Compositions and Formulations

[0218] In some embodiments of the combination therapy methods, compositions, combinations, kits and uses provided herein, the combination therapy can be administered in one or more compositions, e.g., one or more pharmaceutical compositions, containing the combination agents.

[0219] In some embodiments, a composition containing a combination agent can include a carrier, such as a diluent, adjuvant, excipient, or vehicle, with which the combination agent is administered. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin. Such compositions will contain a therapeutically effective amount of the combination agent, generally in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to a patient. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. The pharmaceutical composition may contain any one or more of diluent(s), adjuvant(s), antiadherent(s), binder(s), coating(s), filler(s), flavor(s), color(s), lubricant(s), glidant(s), preservative(s), surfactant(s), adsorbent(s), emulsifier(s), pharmaceutical excipient(s), pH buffer(s), or sweetener(s), and combinations thereof. In some embodiments, the pharmaceutical composition may be in liquid, solid, lyophilized powder, gel form, and / or combinations thereof. In some aspects, the choice of carrier is determined in part by the particular inhibitor and / or method of administration.

[0220] Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citric acid, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; serum These include, but are not limited to, proteins such as albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants, stabilizers, and / or preservatives, such as polyethylene glycol (PEG). The composition containing the combined drug may be lyophilized.

[0221] In some embodiments, the pharmaceutical composition can be formulated for administration by any known route, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical, local, otic, inhalation, buccal (e.g., sublingual), and transdermal administration, or any other route. In some embodiments, other modes of administration are also contemplated. In some embodiments, administration is by bolus injection, injection, e.g., intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, administration is parenteral, intrapulmonary and intranasal, and if desired for local treatment, intralesional administration.Parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.In some embodiments, a given dose is administered by a single bolus administration.In some embodiments, it is administered by multiple bolus administrations over a period of, for example, 3 days or less, or by continuous infusion administration.

[0222] In some embodiments, administration can be local, topical, or systemic depending on the location of treatment. In some embodiments, local administration to the area requiring treatment can be achieved, for example, but not limited to, by local infusion during surgery, topical application in conjunction with a wound dressing after surgery, injection, using a catheter, using a suppository, or using an implant. In some embodiments, the composition can also be administered sequentially, intermittently, or in the same composition with other biologically active agents. In some embodiments, administration can also include controlled release systems, including controlled release formulations and device-controlled release, such as using a pump. In some embodiments, administration is oral.

[0223] In some embodiments, pharmaceutically and therapeutically active compounds and their derivatives are typically formulated and administered in unit or multiple dosage forms. Each unit dosage contains a predetermined amount of the therapeutically active compound sufficient to produce the desired therapeutic effect, in combination with the required pharmaceutical carrier, vehicle, or diluent. In some embodiments, unit dosage forms include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, oral solutions or suspensions, and oil-and-water emulsions, each containing an appropriate amount of the compound or a pharmaceutically acceptable derivative thereof. Unit dosage forms may be contained in ampoules and syringes or individually packaged tablets or capsules. Unit dosage forms may be administered in fractions or multiples thereof. In some embodiments, multiple dosage forms are multiple identical unit dosage forms packaged in a single container to be administered in segregated unit dosage forms. Examples of multiple dosage forms include vials, bottles of tablets or capsules, or bottles of pints or gallons.

[0224] The active ingredient can be entrapped in microcapsules, colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or macroemulsions. In certain embodiments, pharmaceutical compositions containing combination drugs are formulated as inclusion complexes, such as cyclodextrin inclusion complexes, or as liposomes. Liposomes can function to target host cells (e.g., T-cells or NK cells) to specific tissues. Many methods are available for preparing liposomes, such as those described in Szoka et al., Ann. Rev. Biophys. Bioeng., 9: 467 (1980) and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0225] In some embodiments, pharmaceutical compositions containing combination drugs can use time-release, delayed release, and sustained release delivery systems so that the composition is delivered before and in sufficient time to cause sensitization of the treated area. Many types of release delivery systems are available and known. Such systems can avoid repeated administration of the composition, thereby increasing convenience for patients and physicians.

[0226] The composition containing the combined drug may be lyophilized. Depending on the desired route of administration and preparation, the composition may contain auxiliary substances such as wetting, dispersing or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling agents or viscosity-enhancing additives, preservatives, flavoring agents, coloring agents, etc. In some embodiments, standard textbooks can be consulted to prepare suitable preparations.

[0227] Various additives can be added to enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of microbial action can be ensured by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, such as aluminum monostearate and gelatin.

[0228] Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules.

[0229] In some embodiments, the composition containing the combined agent is administered in the form of a salt, for example, a pharmaceutically acceptable salt.Suitable pharmaceutically acceptable acid addition salts include those derived from mineral acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid, and organic acids, such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid and arylsulfonic acids, for example, p-toluenesulfonic acid. 2. Immunomodulatory Compound Dosage Schedule

[0230] In some embodiments, the provided methods of combination therapy involve administering an immunomodulatory compound, e.g., Compound A, and a cell therapy, such as a T cell therapy (e.g., a CAR-expressing T cell).

[0231] In some embodiments, administration of the immunomodulatory compound, e.g., Compound A, is initiated prior to, following, during, during the course of, simultaneously with, near-simultaneously with, sequentially, and / or intermittently with administration of cell therapy, such as T cell therapy (e.g., CAR-expressing T cells). In some embodiments, the methods involve initiating administration of the immunomodulatory compound, e.g., Compound A, prior to administration of T cell therapy. In other embodiments, the methods involve initiating administration of the immunomodulatory compound, e.g., Compound A, after administration of T cell therapy. In some embodiments, the dosing schedule involves initiating administration of the immunomodulatory compound, e.g., Compound A, simultaneously or concurrently with administration of T cell therapy.

[0232] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in cycles. In some embodiments, a cycle includes an administration period during which the immunomodulatory compound, e.g., Compound A, is administered, followed by a rest period during which the immunomodulatory compound, e.g., Compound A, is not administered. In some embodiments, the total number of days in a cycle, e.g., from the beginning of administration of the immunomodulatory compound, e.g., Compound A, is more than or about 21 days, 28 days, 30 days, 40 days, 50 days, 60 days, or more.

[0233] In some embodiments, the initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs in at least one cycle, and the initiation of administration of the T cell therapy occurs on the same day, optionally simultaneously. In some embodiments, the initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs prior to the initiation of administration of the T cell therapy in at least one cycle. In some embodiments, the initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs concurrently or on the same day as the initiation of administration of the T cell therapy in at least one cycle. In some embodiments, the immunomodulatory compound, e.g., Compound A, occurs 0-15 days, 0-6 days, 0-96 hours, 0-24 hours, 0-12 hours, 0-6 hours, or 0-2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours, 6 hours to The immunomodulatory compound, e.g., Compound A, is administered 0 to 30 days or any number thereof, such as 24 hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days to 30 days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered less than about 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour prior to the initiation of T cell therapy.

[0234] In some of any such embodiments in which the immunomodulatory compound, e.g., Compound A, is given prior to cell therapy (e.g., T cell therapy such as CAR-T cell therapy), administration of the immunomodulatory compound, e.g., Compound A, continues at regular intervals until and / or for a time after the initiation of cell therapy.

[0235] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered after or in addition to administration of cell therapy (e.g., T cell therapy such as CAR-T cell therapy). In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered within or about 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 96 hours, 4 days, 5 days, 6 or 7 days, 14 days, 15 days, 21 days, 24 days, 28 days, 30 days, 36 days, 42 days, 60 days, 72 days, or 90 days after initiation of administration of cell therapy (e.g., T cell therapy). In some embodiments, provided methods involve continued administration of the immunomodulatory compound, such as at regular intervals, after initiation of administration of cell therapy.

[0236] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, or up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after the start of administration of the cell therapy (e.g., T cell therapy such as CAR-T cell therapy).

[0237] In some of any such above-described embodiments, the immunomodulatory compound, e.g., Compound A, is administered before and after the initiation of administration of the cell therapy (e.g., T cell therapy, such as CAR-T cell therapy).

[0238] In some embodiments, initiation of administration of an immunomodulatory compound, e.g., Compound A, occurs when: (i) a peak or maximum level of T cell therapy cells is detectable in the subject's blood; (ii) the number of T cell therapy cells detectable in the blood, after becoming detectable in the blood, becomes undetectable or is reduced, optionally reduced compared to an earlier time point after administration of the T cell therapy; (iii) the number of T cell therapy cells detectable in the blood is reduced by more than 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold or more compared to the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of administration of the T cell therapy; (iv) the T At a time point after the peak or maximum level of cell therapy cells is detectable in the subject's blood, the number of T cells or cells derived therefrom detectable in the subject's blood is less than 10%, less than 5%, less than 1%, or less than 0.1% of the total peripheral blood mononuclear cells (PBMCs) in the subject's blood; (v) the subject exhibits disease progression and / or relapse following remission after treatment with T cell therapy, and / or (iv) the subject exhibits an increased tumor burden compared to the tumor burden at a time point before or after administration of the T cells and before initiation of administration of the immunomodulatory compound, as appropriate, performed shortly thereafter or within 1-3 days thereafter.

[0239] In some embodiments, the initiation of administration of the immunomodulatory compound, e.g., Compound A, follows the initiation of administration of T cell therapy for at least one cycle. In some embodiments, the initiation of administration of the immunomodulatory compound, e.g., Compound A, follows at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or about at least 12 days, at least or about at least 14 days, at least or about at least 15 days, at least or about at least 21 days, at least or about 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days, or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after the initiation of administration of T cell therapy. In some embodiments, initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs at least 2 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after initiation of administration of T cell therapy. In some embodiments, initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs 2 to 28 days or 7 to 21 days after initiation of administration of T cell therapy. In some embodiments, initiation of administration of the immunomodulatory compound, e.g., Compound A, occurs more than or about 14, 15, 16, 17, 18, 19, 20, 21, 24, or 28 days after initiation of administration of T cell therapy. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered several times a day, twice a day, daily, every other day, three times a week, twice a week, or once a week after initiation of cell therapy. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered daily. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered twice daily. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered three times daily.In other embodiments, the immunomodulatory compound, e.g., Compound A, is administered every other day. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered daily. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for several consecutive days during the administration period, such as up to about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more than 30 consecutive days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for more than or about 7 consecutive days, more than or about 14 consecutive days, more than or about 21 consecutive days, more than or about 21 consecutive days, or more than or about 28 consecutive days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for up to 21 consecutive days during the administration period. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for up to 21 consecutive days during the administration period, with the cycle comprising more than 30 days beginning after the start of administration of the immunomodulatory compound.

[0240] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 days or less, or for 30 consecutive days or less, during the administration period. In certain embodiments, the immunomodulatory compound, e.g., Compound A, is administered once daily for 14 days over a 21-day treatment cycle. In certain embodiments, the immunomodulatory compound, e.g., Compound A, is administered once daily for 21 days over a 28-day treatment cycle. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered for 14 consecutive days or less, during the administration period.

[0241] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in cycles, where the cycle comprises administration of the immunomodulatory compound, e.g., Compound A, for several consecutive days, followed by a rest period during which the immunomodulatory compound is not administered. In some embodiments, the rest period is greater than about 1 day, greater than about 3 days, greater than about 5 days, greater than about 7 days, greater than about 8 days, greater than about 9 days, greater than about 10 days, greater than about 11 days, greater than about 12 days, greater than about 13 days, greater than about 14 days, greater than about 15 days, greater than about 16 days, greater than about 17 days, greater than about 18 days, greater than about 19 days, greater than about 20 days, or greater than about 21 days or more. In some embodiments, the rest period is greater than 7 days, greater than 14 days, greater than 21 days, or greater than 28 days. In some embodiments, the rest period is greater than about 14 days. In some embodiments, the cycle of administration of the immunomodulatory compound does not contain a rest period.

[0242] In some embodiments, the immunomodulatory compound, for example, Compound A, is administered in cycles, and the cycle is repeated at least once. In some embodiments, the immunomodulatory compound, for example, Compound A, is administered for at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles, at least 7 cycles, at least 8 cycles, at least 9 cycles, at least 10 cycles, at least 11 cycles, or at least 12 cycles. In some embodiments, the immunomodulatory compound, for example, Compound A, is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 cycles.

[0243] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day, once a day, every other day, every third day, twice a week, once a week, or only once a day prior to or following the initiation of administration of T cell therapy. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in multiple doses at regular intervals prior to, during, the course of, and / or after the period of administration of T cell therapy. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in one or more doses at regular intervals prior to administration of T cell therapy. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in one or more doses at regular intervals after administration of T cell therapy. In some embodiments, one or more doses of the immunomodulatory compound, e.g., Compound A, can be administered simultaneously with the administration of the dose of T cell therapy.

[0244] In some embodiments, the dose, frequency, duration timing and / or sequence of administration of an immunomodulatory compound, e.g., Compound A, is determined based on certain thresholds or criteria resulting from the screening steps described herein and / or evaluation of treatment outcomes, e.g., those described in Section III herein.

[0245] In some embodiments, the method involves administering a cell therapy to a subject who has previously been administered a therapeutically effective amount of an immunomodulatory compound. In some embodiments, the immunomodulatory compound is administered to the subject before administering to the subject a dose of cells expressing the recombinant receptor. In some embodiments, treatment with the immunomodulatory compound occurs simultaneously with administration of the dose of cells. In some embodiments, the immunomodulatory compound is administered after administration of the dose of cells.

[0246] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered daily for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more than 21 days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered twice daily for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more than 21 days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered three times daily for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more than 21 days. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered every other day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 days or more than 21 days.

[0247] In some embodiments of the methods provided herein, the immunomodulatory compound, e.g., Compound A, and the T cell therapy are administered simultaneously or near simultaneously.

[0248] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in an amount of 0.1 mg or about 0.1 mg to about 100 mg, 0.1 mg or about 0.1 mg to 50 mg, 0.1 mg or about 0.1 mg to 25 mg, 0.1 mg or about 0.1 mg to 10 mg, 0.1 mg or about 0.1 mg to 5 mg, 0.1 mg or about 0.1 mg to 1 mg, 1 mg or about 1 mg to 100 mg, 1 mg or about 1 mg to 50 mg, 1 mg or about 1 mg to 25 mg, 1 mg or about 1 mg to 10 mg, In some embodiments, the amount is a once-daily dose of an immunomodulatory compound, e.g., Compound A, of 1 mg or about 1 mg to 5 mg, 5 mg or about 5 mg to 100 mg, 5 mg or about 5 mg to 50 mg, 5 mg or about 5 mg to 25 mg, 5 mg or about 5 mg to 10 mg, 10 mg or about 10 mg to 100 mg, 10 mg or about 10 mg to 50 mg, 10 mg or about 10 mg to 25 mg, 25 mg or about 25 mg to 100 mg, 25 mg or about 25 mg to 50 mg, or 50 mg or about 50 mg to 100 mg, inclusive.

[0249] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered at a dosage of about 1 mg to about 20 mg, e.g., about 1 mg to about 10 mg, about 2.5 mg to about 7.5 mg, about 5 mg to about 15 mg, e.g., about 5 mg, 10 mg, 15 mg, or 20 mg. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered at a dose of about 10 μg / kg to 5 mg / kg, e.g., about 100 μg / kg to about 2 mg / kg, about 200 μg / kg to about 1 mg / kg, about 400 μg / kg to about 600 μg / kg, e.g., about 500 μg / kg. In some embodiments, the amount is a once-daily dose of the immunomodulatory compound, e.g., Compound A. In some embodiments, the immunomodulatory compound is Compound A. In some embodiments, the immunomodulatory compound is Compound B. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in a total daily dosage of at least about 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day, or 100 mg per day. In some embodiments, the dose of the immunomodulatory compound, e.g., Compound A, is at or about 25 mg per day. In certain embodiments, the dose of the immunomodulatory compound, e.g., Compound A, is at or about 10 mg per day. In some embodiments, the immunomodulatory compound is Compound A. In some embodiments, the immunomodulatory compound is Compound B.

[0250] In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in an amount greater than or equal to 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, or more than about 25 mg. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered in an amount greater than or equal to 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, or greater than or equal to 15 mg per day, or more than about 25 mg per day. In some embodiments, the immunomodulatory compound is Compound A. In some embodiments, the immunomodulatory compound is Compound B.

[0251] In some embodiments, provided methods include administering to a subject an effective amount of Compound A per day to modulate the activity and / or function of a T cell therapy. In some embodiments, Compound A is administered at a dosage of 0.1 mg or about to 1 mg or about. In some embodiments, the amount is 0.1 mg or about, 0.2 mg or about, 0.3 mg or about, 0.4 mg or about, 0.5 mg or about, 0.6 mg or about, 0.7 mg or about, 0.8 mg or about, 0.9 mg or about, or 1.0 mg or about, or any value between any of the foregoing. In some embodiments, the amount of Compound A is administered in a cycling regimen involving daily administration for a four-week period or three weeks in a cycle. The administration of Compound A is carried out over a period of time, for example, generally for more than one week, for example, for one or more months, two or more months, three or more months, four or more months, five or more months, or six or more months. Exemplary dosing regimens are described herein.

[0252] In some embodiments, the methods provided minimize or avoid toxicity following administration to a subject of a T cell therapy and / or an immunomodulatory compound, e.g., Compound A. In some embodiments, the methods provided herein involve administering a dose substantially lower than the dose identified to be the MTD of the compound.

[0253] In some of the embodiments, the methods and uses include administration of Compound A. In some embodiments, administration of Compound A is initiated after (subsequent to) the initiation of cell therapy, such as T cell therapy (e.g., CAR-expressing T cells). In some embodiments, administration of Compound A is initiated at or after a peak or maximum level of T cell therapy cells is detectable in the subject's blood. In some cases, administration of Compound A is initiated when (i) a peak or maximum level of T cell therapy cells is detectable in the subject's blood; (ii) the number of T cell therapy cells detectable in the blood becomes undetectable or is reduced, optionally compared to an earlier time point after administration of the T cell therapy; (iii) the number of T cell therapy cells detectable in the blood is reduced by 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold, or more, relative to the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of administration of the T cell therapy; (iv (v) the subject exhibits disease progression and / or relapse following remission after treatment with T cell therapy; and / or (iv) the subject exhibits an increased tumor burden compared to the tumor burden prior to or following administration of the cells and prior to initiation of administration of Compound A. In certain embodiments, the provided methods are performed to enhance, increase, or potentiate T cell therapy in a subject in whom a peak response to T cell therapy was observed, but in whom the response, e.g., reduction in T cell presence and / or tumor burden, has become diminished or is no longer detectable.

[0254] In some embodiments, administration of Compound A is initiated at or about 14 days to about 35 days after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is initiated about 21 to about 35 days after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is initiated about 21 to about 28 days after initiation of administration of T cell therapy. In some embodiments, administration of Compound A begins at or about 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, or 35 days or about 35 days after initiation of administration of the T cell therapy.

[0255] In some embodiments, at the time the subject is first administered Compound A and / or at any subsequent time point after initiation of administration, the subject does not exhibit signs or symptoms of severe toxicity, e.g., severe cytokine release syndrome (CRS) or severe toxicity. In some embodiments, Compound A is administered at a time when the subject does not exhibit signs or symptoms of severe CRS and / or does not exhibit Grade 3 or higher CRS, e.g., prolonged Grade 3 CRS or Grade 4 or 5 CRS. In some embodiments, Compound A is administered at a time when the subject does not exhibit signs or symptoms of severe neurotoxicity and / or does not exhibit Grade 3 or higher neurotoxicity, e.g., prolonged Grade 3 neurotoxicity or Grade 4 or 5 neurotoxicity. In some aspects, between the time of initiation of administration of T cell therapy and the time of administration of Compound A, the subject did not exhibit severe CRS and / or did not exhibit Grade 3 or higher CRS, e.g., prolonged Grade 3 CRS or Grade 4 or 5 CRS. In some instances, between the time of initiation of administration of T cell therapy and the time of administration of Compound A, the subject does not experience severe neurotoxicity and / or does not experience grade 3 or higher neurotoxicity, e.g., prolonged grade 3 neurotoxicity or grade 4 or 5 neurotoxicity.

[0256] In some embodiments, the daily dose of Compound A is in an amount of 0.1 mg to 5 mg or an approximation thereof. In some embodiments, the daily dose of Compound A is in an amount of about 0.1 mg to about 5 mg, about 0.5 mg to about 5 mg, about 1 mg to about 5 mg, about 1.5 mg to about 5 mg, about 2 mg to about 5 mg, about 2.5 mg to about 5 mg, about 3 mg to about 5 mg, about 0.1 mg to about 4 mg, about 0.1 mg to about 4 mg, about 1 mg to about 4 mg, about 1.5 mg to about 4 mg, about 2 mg to about 4 mg, about 2.5 mg to about 4 mg, about 3 mg to about 4 mg, about 0.1 mg to about 3.5 mg, about 0.5 mg to about 3.5 mg, about 1 mg to about 3.5 mg, about 1.5 mg to about 3.5 mg, about 2 mg to about 3.5 mg, about 2.5 mg to about 3.5 mg. g, about 3 mg to about 3.5 mg, about 0.1 mg to about 3 mg, about 0.5 mg to about 3 mg, about 1 mg to about 3 mg, about 1.5 mg to about 3 mg, about 2 mg to about 3 mg, about 2.5 mg to about 3 mg, about 0.1 mg to about 2.5 mg, about 0.5 mg to about 2.5 mg, about 1 mg to about 2.5 mg, about 1.5 mg to about 2.5 mg, about 2 mg to about 2.5 mg, about 0.1 mg to about 2 mg, about 0.5 mg to about 2 mg, about 1 mg to about 2 mg, about 1.5 mg to about 2 mg, about 0.1 mg to about 1.5 mg, about 0.5 mg to about 1.5 mg, about 1 mg to about 1.5 mg, about 0.1 mg to about 1 mg, or about 0.5 mg to about 1 mg.

[0257] In some embodiments, the dose of Compound A per day it is administered is about or at least about 0.5 mg, or an amount equal to or at least that value. In some embodiments, the dose of Compound A per day it is administered is about or at least about 1 mg, or an amount equal to or at least that value. In some embodiments, the dose of Compound A per day it is administered is about or at least about 1.5 mg, or an amount equal to or at least that value. In some embodiments, the dose of Compound A per day it is administered is about or at least about 2 mg, or an amount equal to or at least that value. In some embodiments, the dose of Compound A per day it is administered is about or at least about 2.5 mg, or an amount equal to or at least that value. In some embodiments, the dose of Compound A per day it is administered is about or at least about 3 mg, or an amount equal to or at least that value. In some of any such embodiments, the dose of Compound A per day it is administered is about 5 mg or less. In some embodiments, Compound A is administered in an amount of about 4.5 mg or less per day. In some embodiments, Compound A is administered in an amount of about 4 mg or less per day. In some embodiments, Compound A is administered in an amount of about 3.5 mg or less per day. In some embodiments, Compound A is administered in an amount of about 3 mg or less per day. In some embodiments, Compound A is administered in an amount of about 2.5 mg or less per day. In some embodiments, Compound A is administered in an amount of about 2 mg or less per day. In some embodiments, Compound A is administered in an amount of about 1.5 mg or less per day. In some embodiments, Compound A is administered in an amount of about 1 mg or less per day.

[0258] In some embodiments, the dose of Compound A per day it is administered is at or about 3 mg. In some embodiments, the dose of Compound A per day it is administered is at or about 2.5 mg. In some embodiments, the dose of Compound A per day it is administered is at or about 2 mg. In some embodiments, the dose of Compound A per day it is administered is at or about 1.5 mg. In some embodiments, the dose of Compound A per day it is administered is at or about 1 mg per day.

[0259] In some embodiments, compound A may be present in an amount of about 10 nM to about 500 nM, about 40 nM to about 500 nM, about 60 nM to about 500 nM, about 80 nM to about 500 nM, about 100 nM to about 500 nM, about 150 nM to about 500 nM, about 200 nM to about 500 nM, about 250 nM to about 500 nM, about 300 nM to about 500 nM, about 350 nM to about 500 nM, about 400 nM to about 500 nM, 10 nM to about 400 nM, about 40 nM to about 400 nM, about 60 nM to about 400 nM, about 80 nM to about 400 nM, about 100 nM to about 500 nM, M ~ about 400nM, about 150nM - about 400nM, about 200nM - about 400nM, about 250nM - about 400nM, about 300nM - about 400nM, about 350nM - about 400nM, 10nM - about 350nM, about 40nM - about 350nM, about 60nM - about 350nM, about 80nM to about 350nM, about 100nM to about 350nM, about 150nM to about 350nM, about 200nM to about 350nM, about 250nM to about 350nM, about 300nM to about 350nM, about 10nM to about 300nM, about 40nM to about 30 0nM, approximately 60nM to approximately 300nM, approximately 80nM to approximately 300nM, approximately 100nM to approximately 300nM, approximately 150nM to approximately 300nM, approximately 200nM to approximately 300nM, approximately 250nM to approximately 250nM, approximately 10nM to approximately 250nM, approximately 40nM to approximately 250nM , about 60nM to about 250nM, about 80nM to about 250nM, about 100nM to about 250nM, about 150nM to about 250nM, about 200nM to about 250nM, about 10nM to about 200nM, about 40nM to about 200nM, about 60nM to about 200nM, about 80 and a maximum concentration of Compound A in the blood (C), such as for each week of the cycling regimen or for at least one week of the cycling regimen, in the range of about 100 nM to about 200 nM, about 100 nM to about 200 nM, about 150 nM to about 200 nM, about 10 nM to about 150 nM, about 40 nM to about 150 nM, about 60 nM to about 150 nM, about 80 nM to about 150 nM, about 100 nM to about 150 nM, about 10 nM to about 100 nM, about 40 nM to about 100 nM, about 60 nM to about 100 nM, or about 80 nM to about 100 nM. max In some embodiments, Compound A is administered in an amount that achieves a C within the range for at least about 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 16 hours, or 24 hours. maxis administered in a dose that maintains

[0260] In some embodiments, Compound A has a C of Compound A in the blood of about or at least about 40 nM. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood of about or at least about 60 nM. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood of about or at least about 80 nM, such as over each week of the cycling regimen or over at least one week of the cycling regimen. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as about or at least about 90 nM over each week of the cycling regimen or over at least one week of the cycling regimen. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as about or at least about 100 nM over each week of the cycling regimen or over at least one week of the cycling regimen. max In some embodiments, Compound A is administered in an amount that achieves C for at least about 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 16 hours, or 24 hours. max is administered in a dose that maintains

[0261] In some embodiments, Compound A provides a C of Compound A in the blood, such as over each week of the cycling regimen or over at least one week of the cycling regimen, of about 500 nM or less. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 400 nM or less. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 350 nM or less. maxIn some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 300 nM or less. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 250 nM or less. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 200 nM or less. max In some embodiments, Compound A is administered in an amount that achieves a C of Compound A in the blood, such as for each week of the cycling regimen or for at least one week of the cycling regimen, of about 150 nM or less. max is administered in an amount that achieves

[0262] In some embodiments, Compound A is administered in a cycling regimen involving repeated dosing of the compound over a specified period or duration. In some embodiments, Compound A is administered in a cycling regimen, where the compound is administered in an effective amount, such as the amounts described above, for each week of the cycling regimen or for at least one week of the cycling regimen, for each of five or fewer days per week for a period of more than one week. In some embodiments, the amount of Compound A per administration or per day is 3 mg or less (e.g., 3 mg, 2.5 mg, 2 mg, 1.5 mg, 1 mg, 0.5 mg or less). In some embodiments, the amount of Compound A per administration or per day is 3 mg or less, 2.5 mg or less, 2 mg or less, 1.5 mg or less, 1 mg or less, 0.5 mg or less. In some embodiments, the amount of Compound A per administration or per day is about 1 mg to about 2 mg (e.g., 1 mg or less, 2 mg or less).

[0263] In some embodiments, each week of the cycling regimen comprises administering Compound A for no more than 5 days each week. In some embodiments, each week of the cycling regimen comprises administering Compound A for no more than 4 days each week. In some embodiments, each week of the cycling regimen comprises administering Compound A for no more than 3 days each week.

[0264] In some embodiments, each week of the cycling regimen comprises administering Compound A for 3-5 days per week. In some embodiments, each week of the cycling regimen comprises administering Compound A for 4-5 days per week. In some embodiments, each week of the cycling regimen comprises administering Compound A for 3-4 days per week.

[0265] In some embodiments, each week of the cycling regimen or at least one week of the cycling regimen comprises administering Compound A for no more than five consecutive days per week, followed by a rest period during the remainder of the week during which the compound is not administered. In some embodiments, each week of the cycling regimen or at least one week of the cycling regimen comprises administering Compound A for three to five consecutive days per week, followed by a rest period during the remainder of the week during which the compound is not administered. In some embodiments, each week of the cycling regimen or at least one week of the cycling regimen comprises administering Compound A for three consecutive days per week, followed by a four-day rest period during which the compound is not administered. In some embodiments, each week of the cycling regimen or at least one week of the cycling regimen comprises administering Compound A for four consecutive days per week, followed by a three-day rest period during which the compound is not administered. In some embodiments, each week of the cycling regimen or at least one week of the cycling regimen comprises administering Compound A for five consecutive days per week, followed by a two-day rest period during which the compound is not administered.

[0266] In some embodiments, the cycling regimen for administering Compound A is carried out for a period subsequent to the initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of more than one week after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of about or at least about one month after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of about or at least about two months after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of about or at least about three months after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of about or at least about four months after initiation of administration of T cell therapy. In some embodiments, administration of Compound A is extended for a period of about or at least about five months after initiation of administration of T cell therapy.

[0267] In some embodiments, administration of Compound A extends over a period of at least 3 months. In some embodiments, administration of Compound A extends over a period of 90 days or so, 100 days or so, 105 days or so, 110 days or so, 115 days or so, 120 days or so, 125 days or so, 130 days or so, 135 days or so, 140 days or so, 145 days or so, 150 days or so, 155 days or so, 160 days or so, 165 days or so, 170 days or so, 175 days or so, 180 days or so, 185 days or so, 190 days or so, 195 days or so, 200 days or so or more after initiation of administration of T cell therapy.

[0268] In some embodiments, administration of Compound A extends for a period of 90 days or about 90 days or 3 months or about 3 months after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, administration of Compound A extends for a period of 120 days or 4 months or about 120 days after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, administration of Compound A extends for a period of 150 days or 5 months or about 150 days after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, administration of Compound A extends for a period of 180 days or 6 months or about 180 days after initiation of administration of T cell therapy (e.g., CAR T cell therapy).

[0269] In some embodiments, if the subject achieves a complete response (CR) following treatment before or about 6 months, or if the cancer (e.g., B-cell malignancy) progresses or recurs following remission following treatment, administration of Compound A is terminated or discontinued at the end of the period (e.g., 3, 4, 5, or 6 months or about) after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, the period is of fixed duration such that administration of Compound A continues for the period, even if the subject achieves a complete response (CR) at a time point before the end of the period. In some embodiments, the subject has a CR with minimal residual disease (MRD). In some embodiments, the subject has a CR that is MRD-.

[0270] In some embodiments, if the subject exhibits a partial response (PR) or stable disease (SD) after treatment, administration of Compound A is continued after the end of the period, for example, for more than 3, 4, 5, or 6 months or about 3, 4, 5, or 6 months after initiation of administration of T cell therapy (e.g., CAR T cells). In some embodiments, administration of Compound A is continued for more than 6 months after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, for subjects who exhibit PR or SD at the end of the initial period, administration of Compound A is continued until the subject achieves a complete response (CR) following treatment, or until the cancer (e.g., multiple myeloma, e.g., relapsed / refractory multiple myeloma) progresses or recurs following remission after treatment.

[0271] In some embodiments, administration of Compound A is carried out in a cycling regimen comprising administering Compound A in an amount of about 3 mg or less (e.g., 1-3 mg, 1 mg, 2 mg, or 3 mg) per day for 5 days or less (e.g., 3, 4, or 5 days) per week for a period of more than 1 week. In some embodiments, each week of the cycling regimen involves administration of the compound for 3, 4, or 5 consecutive days, followed by a rest period during the remainder of the week during which the compound is not administered. In some embodiments, each week of the cycling regimen involves administration of the compound for 5 days, followed by a 2-day rest period during which the compound is not administered. In some embodiments, administration of Compound A is initiated more than about 14 to about 35 days (e.g., about 21 to about 35 days) after initiation of administration of the cell therapy. In some embodiments, at the time of administration of Compound A, the subject does not exhibit severe toxicity following administration of T cell therapy (e.g., CAR T cells).

[0272] In some embodiments, administration of Compound A is carried out in a cycling regimen comprising administering an effective amount of Compound A for no more than 5 days per week (e.g., 3, 4, or 5 days) for a period extending for 3 or more months, 4 or more months, 5 or more months, or 6 or more months after initiation of administration of cell therapy (e.g., T cell therapy). In some embodiments, the period extends for 3 or more months, 4 or more months, 5 or more months, or 6 or more months. In some embodiments, each week of the cycling regimen involves administration of the compound for each of 3, 4, or 5 consecutive days, followed by a rest period for the remainder of the week during which the compound is not administered. In some embodiments, each week of the cycling regimen involves administration of the compound for each of 5 consecutive days, followed by a 2-day rest period during which the compound is not administered. In some embodiments, administration of Compound A is initiated more than about 14 to about 35 days (e.g., about 21 to about 35 days, e.g., 28 days or any number thereof) after initiation of administration of cell therapy. In some embodiments, at the time of administration of Compound A, the subject does not experience severe toxicity following administration of cell therapy. In some embodiments, administration of Compound A is terminated or discontinued if the subject achieves a complete response (CR) following treatment or the cancer, e.g., B-cell malignancy, progresses or recurs following remission following treatment before or about the end of the period. In some embodiments, administration of Compound A is continued for the period, even if the subject achieves a complete response (CR) at a time point before the end of the period. In some embodiments, after initiation of administration of T cell therapy, administration of Compound A is continued after the end of the initial period if the subject experiences a partial response (PR) or stable disease (SD) following treatment. In some embodiments, administration of Compound A is repeated until the subject achieves a complete response (CR) following treatment, or until the cancer, e.g., multiple myeloma, e.g., relapsed / refractory multiple myeloma, progresses or relapses following remission after treatment.

[0273] In some embodiments, administration of Compound A is carried out in a cycling regimen comprising administering Compound A in an amount of about 3 mg or less per day (e.g., 1-3 mg, 1 mg, 2 mg, or 3 mg) for each of five days or less (e.g., three, four, or five days) per week for a period of about three months or more (e.g., for a period of three, four, five, or six months or so) after initiation of administration of T cell therapy (e.g., CAR T cell therapy). In some embodiments, each week of the cycling regimen involves administration of the compound for each of three, four, or five consecutive days, followed by a rest period for the remainder of the week during which the compound is not administered. In some embodiments, each week of the cycling regimen involves administration of the compound for each of five days, followed by a two-day rest period during which the compound is not administered. In some embodiments, administration of Compound A is initiated more than about 14 to about 35 days (e.g., about 21 to about 35 days, e.g., 28 days or so) after initiation of administration of the cell therapy. In some embodiments, at the time of administering Compound A, the subject does not experience severe toxicity following administration of cell therapy. In some embodiments, the cancer is multiple myeloma, e.g., relapsed / refractory multiple myeloma. In some embodiments, if the subject achieves a complete response (CR) following treatment before or about six months, or if the cancer, e.g., B-cell malignancy, progresses or recurs following remission after treatment, administration of Compound A is terminated or discontinued at or about six months after initiation of administration of T cell therapy. In some embodiments, even if the subject achieves a complete response (CR) at a time point before the end of the period, the cycling regimen is continued for the entire period. In some embodiments, if the subject experiences a partial response (PR) or stable disease (SD) following treatment at or about six months, administration of Compound A is continued further after the end of the period, e.g., for more than six months after initiation of administration of cell therapy. In some embodiments, administration of Compound A is continued until the subject achieves a complete response (CR) following treatment, or until the cancer, e.g., multiple myeloma, e.g., relapsed / refractory multiple myeloma, progresses or relapses following remission after treatment.

[0274] In some embodiments, administration of Compound A is carried out in a cycling regimen comprising administering Compound A in an amount of about 1 mg to about 3 mg (e.g., 1 mg, 2 mg, or 3 mg) per day for each of five consecutive days per week for at or about six months or more following initiation of cell therapy (e.g., T cell therapy), followed by a two-day rest period during which no compound is administered. In some embodiments, administration of Compound A is initiated more than about 14 to about 35 days (e.g., about 21 to about 35 days, e.g., at or about 28 days) after initiation of cell therapy administration. In some embodiments, at the time of administration of Compound A, the subject does not exhibit severe toxicity following administration of cell therapy. In some embodiments, if the subject achieves a complete response (CR) following treatment at or about six months, or if the cancer, e.g., B-cell malignancy, progresses or recurs following remission following treatment, administration of Compound A is discontinued at or about six months after initiation of cell therapy administration. In some embodiments, even if the subject achieves a complete response (CR) at or about 6 months, administration of Compound A is continued for a period of time. In some embodiments, if the subject exhibits a partial response (PR) or stable disease (SD) after treatment at or about 6 months, administration of Compound A is further administered for more than 6 months after initiation of administration of T cell therapy. In some embodiments, administration is continued until the subject achieves a complete response (CR) following treatment, or until the disease or condition, e.g., multiple myeloma, e.g., relapsed / refractory multiple myeloma, progresses or relapses following remission after treatment.

[0275] In some cases, the cycling regimen can be interrupted at any time and / or for one or more periods. In some cases, if a subject develops one or more adverse events, dose-limiting toxicity (DLT), neutropenia or febrile neutropenia, thrombocytopenia, cytokine release syndrome (CRS) and / or neurotoxicity (NT), such as those described in Section IV, the cycling regimen is interrupted or modified. In some embodiments, the amount of Compound A per administration or per day on a particular day of a week is modified after a subject develops one or more adverse events, dose-limiting toxicity (DLT), neutropenia or febrile neutropenia, thrombocytopenia, cytokine release syndrome (CRS) and / or neurotoxicity (NT).

[0276] In any of the above-described embodiments, the immunomodulatory compound, e.g., Compound A, can be administered orally. In some embodiments, the immunomodulatory compound, e.g., Compound A, is administered as a tablet or capsule.

[0277] In some embodiments, the dosage, e.g., the daily dosage, is administered in one or more divided doses, e.g., 2, 3, or 4 doses, or in a single formulation. The immunomodulatory compound, e.g., Compound A, can be administered alone, in the presence of a pharmaceutically acceptable carrier, or in the presence of other therapeutic agents.

[0278] It is understood that higher or lower dosages of the immunomodulatory compound may be used, for example, depending on the particular agent and route of administration. In some embodiments, the immunomodulatory compound can be administered alone or in the form of a pharmaceutical composition, where the compound is in admixture or mixture with one or more pharmaceutically acceptable carriers, excipients, or diluents. In some embodiments, the immunomodulatory compound can be administered systemically or locally to the organ or tissue to be treated. Exemplary routes of administration include, but are not limited to, topical, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intratumoral, and intravenous), oral, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation routes. In some embodiments, the route of administration is oral, parenteral, rectal, nasal, topical, or ocular, or by inhalation. In some embodiments, the immunomodulatory compound is administered orally. In some embodiments, the immunomodulatory compound is administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions.

[0279] Once the patient's disease improves, the dosage can be adjusted. For example, the dosage or frequency of administration, or both, can be reduced as a function of symptoms to a level at which the desired therapeutic or preventive effect is maintained. When the symptoms are alleviated to an appropriate level, treatment can be stopped. However, the patient may require intermittent treatment on a long-term basis after any recurrence of symptoms. The patient may also require chronic treatment on a long-term basis. 3. Dexamethasone Dosage Schedule

[0280] In some embodiments, provided methods of combination therapy involve administering dexamethasone to a subject. In some embodiments, provided methods of combination therapy involve administering dexamethasone and an immunomodulatory compound (e.g., Compound A) to a subject in addition to administering an immunotherapy, e.g., a T cell therapy.

[0281] Relative to administration of immunotherapy, e.g., T cell therapy, dexamethasone administration can begin at any of the times described in Section IB-2. In some embodiments, dexamethasone is administered concurrently with an immunomodulatory compound (e.g., Compound A). In some embodiments, dexamethasone administration begins at the same time that administration of an immunomodulatory compound (e.g., Compound A) begins.

[0282] In some embodiments, dexamethasone is administered at any frequency described in Section IB-2.

[0283] In some embodiments, dexamethasone is administered for any duration described in Section IB-2.

[0284] It will be within the skill of the prescribing physician to provide the recommended dexamethasone dose for treatment.In some embodiments, low-dose dexamethasone treatment is administered.The exemplary dose and treatment cycle for dexamethasone (for example, for low-dose dexamethasone treatment) are described in WO2012 / 078492;Siegel et al. (2020), Br J Haematol 188(4):501-519;San Miguel et al. (2013), Lancet Oncol 14(11):1055-1066;Dimopoulos et al. (2016), Blood 128(4):497-503;Hanaizi et al. (2015), Oncologist 20(3):329-34;And Weisel et al. (2015), Clin Lymphoma Myeloma Leuk 15(9):519-30.

[0285] In some embodiments, administration of dexamethasone is discontinued or adjusted based on any of the criteria described in Section IB-2. 4. Standard triplet regimen

[0286] In some embodiments, provided methods of combination therapy involve administering a standard triplet regimen to a subject in combination with the administration of immunotherapy, e.g., T cell therapy. In some embodiments, the standard triplet regimen is a treatment regimen that is the standard for the treatment of multiple myeloma, e.g., R / R multiple myeloma. Exemplary regimens and combination agents to be administered as part of a standard triplet regimen are known in the art and include those described in Kawano et al. (2015), Immunol Rev 263(1):160-72; Krejcik et al. (2016), Blood 128(3):384-94; Quach et al. (2010), Leukemia 2010 24(1):22-32; Dimopoulos et al. (2020), Leukemia; Abdallah et al. (2018), Blood 132 (Supplement 1): 5642; and Lacy et al. (2014), Blood 124(21): 304.

[0287] In some embodiments, the standard triplet regimen includes administration of daratumumab, pomalidomide, and dexamethasone (e.g., the standard triplet regimen is a DPd regimen). In some embodiments, the combination agents for DPd can be administered in a 28-day cycle. In some embodiments, daratumumab is administered at or about 16 mg / kg. In some embodiments, daratumumab is administered no more than once a week. In some embodiments, pomalidomide is administered at or about a 4 mg dose. In some embodiments, pomalidomide is administered on days 1-21 of a 28-day cycle. In some embodiments, dexamethasone is administered at a low dose. In some embodiments, dexamethasone is administered weekly. In some embodiments, dexamethasone is administered at or about a 40 mg dose. It would be within the skill of a prescribing physician to provide a recommended standard triplet regimen for treatment in combination with immunotherapy, e.g., T-cell therapy.

[0288] In some embodiments, the standard triplet regimen includes administration of pomalidomide, bortezomib, and dexamethasone (e.g., the standard triplet regimen is a PVd regimen). In some embodiments, the combination agent for PVd can be administered in a 21-day or 28-day cycle. In some embodiments, pomalidomide is administered at or about a 4 mg dose. In some embodiments, pomalidomide is administered on days 1-14 of a 21-day cycle. In some embodiments, pomalidomide is administered on days 1-21 of a 28-day cycle. In some embodiments, bortezomib is administered at a dose of 1.3 mg / m or less twice weekly. 2 In some embodiments, dexamethasone is administered at or about 1 mg / kg. In some embodiments, dexamethasone is administered at a low dose. In some embodiments, dexamethasone is administered at a dose of 20 mg or 40 mg. In some embodiments, dexamethasone is administered no more than twice a week. It would be within the skill of the prescribing physician to provide a recommended standard triplet regimen for treatment in combination with immunotherapy, e.g., T cell therapy.

[0289] Compared to administration of immunotherapy, e.g., T cell therapy, administration of the standard triplet regimen can begin at any of the time points described in Section IB-2.

[0290] In some embodiments, administration of the standard triplet regimen is initiated prior to administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated 0-30 days, 0-15 days, 0-6 days, 0-96 hours, 0-24 hours, 0-12 hours, 0-6 hours, 0-2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 6 hours ... In some embodiments, administration of the standard triplet regimen is initiated about 96 hours, 6 hours to 24 hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days to 30 days prior to administration of the immunotherapy, e.g., T cell therapy, or approximately any of these.

[0291] In some embodiments, the standard triplet regimen is administered concurrently with the administration of immunotherapy, e.g., T cell therapy.

[0292] In some embodiments, administration of the standard triplet regimen is initiated following administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 720 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 360 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 240 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 120 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 90 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated up to or up to about 60 days after immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated at or about 56 days after immunotherapy, e.g., T cell therapy. In some embodiments, immunotherapy, e.g., T cell therapy, is administered on day 1 of any of the provided methods, and administration of the standard triplet regimen begins on or about month 3, day 1 (M3D1) of any of the provided methods.

[0293] In some embodiments, administration of the standard triplet regimen begins at least or about at least one day after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least two days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least three days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least four days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least five days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least six days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least seven days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 8 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 9 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 10 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 12 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 14 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 15 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 21 days after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen begins at least or about at least 24 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 28 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 30 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 35 days after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins at least or about at least 42 days after administration of the immunotherapy, e.g., T cell therapy.

[0294] In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 90 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 1 day and 60 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 2 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 2 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 2 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 2 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 2 days and 90 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 2 days and 60 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 90 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 3 days and 60 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 4 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 4 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 4 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 4 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 4 days and 90 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 4 days and 60 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 90 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 5 days and 60 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 6 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 6 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 6 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 6 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 6 and 90 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 6 and 60 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 90 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 7 days and 60 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 14 days and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 14 days and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 14 days and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 14 days and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 14 and 90 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 14 and 60 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 21 and 360 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 21 and 240 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 21 and 180 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 21 and 120 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 21 and 90 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 21 and 60 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 28 and 360 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 28 and 240 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 28 and 180 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 28 and 120 days, or any round number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 28 days and 90 days, or any number therebetween, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 28 days and 60 days, or any number therebetween, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 35 days and 360 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 35 days and 240 days, or any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 35 days and 18 ... Administration of the standard triplet regimen is initiated between 35 days and 120 days, inclusive, or any round number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 35 days and 90 days, inclusive, or any round number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 35 days and 60 days, inclusive, or any round number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 days and 360 days, inclusive, or any round number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 days and 240 days, inclusive, or any round number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 and 180 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 and 120 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 and 90 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 42 and 60 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 49 and 360 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 49 and 240 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen begins between 49 and 180 days, or about any number thereof, inclusive, after administration of the immunotherapy, e.g., T cell therapy.In some embodiments, administration of the standard triplet regimen is initiated between 49 days and 120 days, inclusive, or any number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 49 days and 90 days, inclusive, or any number thereof, after administration of the immunotherapy, e.g., T cell therapy. In some embodiments, administration of the standard triplet regimen is initiated between 49 days and 60 days, inclusive, or any number thereof, after administration of the immunotherapy, e.g., T cell therapy.

[0295] In some embodiments, administration of the standard triplet regimen begins on or about day 56 after the immunotherapy, e.g., T cell therapy. In some embodiments, the immunotherapy, e.g., immunotherapy, e.g., T cell therapy, is administered on day 1 of any of the provided methods, and administration of the standard triplet regimen begins on or about month 3, day 1 (M3D1) of any of the provided methods.

[0296] In some embodiments, the standard triplet regimen is administered for any duration described in Section IB-2.

[0297] In some embodiments, administration of the standard triplet regimen is discontinued or adjusted based on any of the criteria described in Section IB-2. C. Lymphocyte Depletion Treatment

[0298] In some aspects, the provided methods can further include administering one or more lymphodepleting therapies, such as prior to or concurrently with the initiation of administration of T cell therapy. In some embodiments, the lymphodepleting therapy can include administration of a phosphamide, such as cyclophosphamide. In some embodiments, the lymphodepleting therapy can include administration of fludarabine.

[0299] In some embodiments, preconditioning a subject with immunodepleting (e.g., lymphodepleting) therapy can improve the efficacy of adoptive cell therapy (ACT). Preconditioning with lymphodepleting agents, including a combination of cyclosporine and fludarabine, has been effective in improving the efficacy of transferred tumor-infiltrating lymphocyte (TIL) cells in cell therapy, including improving the response and / or persistence of transferred cells. See, for example, Dudley et al., Science, 298, 850-54 (2002); Rosenberg et al., Clin Cancer Res, 17(13):4550-4557 (2011). Similarly, in the context of CAR+T cells, some studies have incorporated lymphodepleting agents, most commonly cyclophosphamide, fludarabine, bendamustine, or a combination thereof, sometimes accompanied by low-dose irradiation. Han et al. Journal of Hematology & Oncology, 6:47 (2013);Kochenderfer et al., Blood, 119: 2709-2720 (2012);Kalos et al., Sci Transl Med, 3(95):95ra73 (2011);Clinical Trial Study Record Nos.: NCT02315612; Please refer to NCT01822652.

[0300] Such preconditioning can be performed with the goal of reducing the risk of one or more of a variety of outcomes that can attenuate the effectiveness of therapy. These include the phenomenon known as "cytokine sink," in which T cells, B cells, and NK cells compete with TILs for homeostatic and activating cytokines such as IL-2, IL-7, and / or IL-15; suppression of TILs by regulatory T cells, NK cells, or other cells of the immune system; and the influence of negative regulators in the tumor microenvironment. Muranski et al., Nat Clin Pract Oncol. December; 3(12): 668-681 (2006).

[0301] Thus, in some embodiments, the provided methods further involve administering a lymphodepleting therapy to the subject. In some embodiments, the methods involve administering the lymphodepleting therapy to the subject prior to administration of the dose of cells. In some embodiments, the lymphodepleting therapy comprises a chemotherapeutic agent, such as fludarabine and / or cyclophosphamide. In some embodiments, administration of the cells and / or lymphodepleting therapy is performed by exogenous delivery.

[0302] In some embodiments, the method includes administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, to the subject, such as cyclophosphamide, fludarabine, or a combination thereof, before administering the dose of cells. For example, the subject can be administered the preconditioning agent at least 2 days, for example, at least 3, 4, 5, 6, or 7 days, before the first or subsequent dose. In some embodiments, the subject is administered the preconditioning agent 7 days or less, for example, 6, 5, 4, 3, or 2 days or less, before administering the dose of cells.

[0303] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose of between or about 20 mg / kg and 100 mg / kg, such as between or about 40 mg / kg and 80 mg / kg. In some aspects, the subject is preconditioned with cyclophosphamide at or about 60 mg / kg. In some embodiments, fludarabine can be administered in a single dose, or in multiple doses, such as daily, every other day, or every third day. In some embodiments, cyclophosphamide is administered once daily for 1 or 2 days.

[0304] In some embodiments, when the lymphodepleting agent comprises fludarabine, the subject receives 10 mg / m 2 ~75mg / m 2 , 15 mg / m 2 ~50mg / m 2 , 20 mg / m 2 ~30mg / m2 or 24 mg / m 2 ~26mg / m 2 or between 1 mg / m 2 ~100mg / m 2 In some instances, the subject receives fludarabine at a dose of between 25 mg / m 2 In some embodiments, fludarabine is administered daily for 1 to 5 days, such as 3 to 5 days. In some embodiments, fludarabine can be administered in a single dose or in multiple doses, for example, every day, every other day, or every third day. In some embodiments, fludarabine is administered daily for 1 to 5 days, such as 3 to 5 days.

[0305] In some embodiments, the lymphodepleting agent comprises a combination of drugs, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of drugs can include cyclophosphamide at any dose or administration schedule, such as those described above, and fludarabine at any dose or administration schedule, such as those described above. For example, in some aspects, the subject receives 60 mg / kg (approximately 2 g / m) of cyclophosphamide prior to the dose of cells. 2 ) of cyclophosphamide and 3 to 5 doses of 25 mg / m 2 Fludarabine is administered.

[0306] In some embodiments, administration of a preconditioning agent prior to injection of a dose of cells improves the outcome of the treatment. For example, in some aspects, preconditioning improves the efficacy of the dose or increases the persistence of recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. In some embodiments, preconditioning treatment increases disease-free survival, such as the percentage of subjects who are alive and show no minimal residual or molecularly detectable disease after a given period following a dose of cells. In some embodiments, the time to median disease-free survival is increased.

[0307] Once the cells are administered to a subject (e.g., a human), the biological activity of the engineered cell population is, in some embodiments, measured by any of a number of known methods. Parameters for evaluation include specific binding of engineered or natural T cells or other immune cells to antigens in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells can also be measured by assaying the expression and / or secretion of certain cytokines, such as CD107a, IFNγ, IL-2, and TNF. In some embodiments, biological activity is measured by assessing clinical outcomes such as reduction in tumor burden or burden, hi some embodiments, toxicity outcomes, cell persistence and / or expansion, and / or the presence or absence of a host immune response are assessed.

[0308] In some embodiments, administration of a preconditioning agent prior to injection of a dose of cells improves the outcome of the treatment, such as by improving the efficacy of the dose, or increases the persistence of the recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. Thus, in some embodiments, the dose of preconditioning agent given in a method that is a combination therapy with a combination agent and cell therapy is higher than the dose given in a method that does not use a combination agent. II. Cellular Therapy and Cell Engineering

[0309] In some embodiments, cellular immunotherapy, such as T cell therapy, for use in conjunction with the provided combination therapy methods comprises administering engineered cells expressing a recombinant receptor designed to recognize and / or specifically bind to a molecule associated with a disease or condition, e.g., cancer, e.g., multiple myeloma, e.g., relapsed or refractory multiple myeloma. In some embodiments, binding to an antigen results in a response, such as an immune response, to such a molecule after binding to such a molecule. In some embodiments, the cells contain or are engineered to contain an engineered receptor, e.g., an engineered antigen receptor, e.g., a chimeric antigen receptor (CAR) or T cell receptor (TCR). Recombinant receptors, such as CARs, generally comprise an extracellular antigen (or ligand) binding domain directed against BCMA, linked, in some aspects, via a linker and / or transmembrane domain(s), to one or more intracellular signaling components. In some aspects, the engineered cells are provided in pharmaceutical compositions and formulations suitable for administration to a subject, such as for adoptive cell therapy. Methods of therapy for administering the cells and compositions to a subject, e.g., a patient, are also provided.

[0310] In some embodiments, the cells contain one or more nucleic acids that have been introduced by genetic engineering, thereby expressing recombinant or engineered products of such nucleic acids. In some embodiments, gene transfer is achieved by first stimulating the cells, e.g., in combination with a stimulus that induces a response such as proliferation, survival, and / or activation, e.g., as measured by expression of cytokines or activation markers, and then transducing and expanding the activated cells in culture to numbers sufficient for clinical application. A. Recombinant Receptors, e.g., Chimeric Antigen Receptors (CARs)

[0311] The cells generally express recombinant receptors, such as antigen receptors, including functional non-TCR antigen receptors, e.g., chimeric antigen receptors (CARs), and other antigen-binding receptors, such as transgenic T cell receptors (TCRs), and other chimeric receptors, among others.

[0312] In some embodiments of the provided methods and uses, engineered cells, such as T cells, express a chimeric receptor, such as a chimeric antigen receptor (CAR), that contains one or more domains that combine a ligand-binding domain (e.g., derived from an antibody or antibody fragment) that provides specificity for a desired antigen (e.g., a tumor antigen) with an intracellular signaling domain. In some embodiments, the intracellular signaling domain is an activation intracellular domain portion, such as a T cell activation domain, that provides a primary activation signal. In some embodiments, the intracellular signaling domain contains, or additionally contains, a costimulatory signaling domain to facilitate effector function. After specific binding to a molecule, e.g., an antigen, the receptor generally delivers an immunostimulatory signal, such as an ITAM transduction signal, to the cell, thereby promoting an immune response targeted to the disease or condition. In some embodiments, the chimeric receptor, when engineered into an immune cell, can modulate T cell activity, and in some cases, modulate T cell differentiation or homeostasis, resulting in engineered cells with improved longevity, survival, and / or persistence in vivo, such as for use in adoptive cell therapy methods.

[0313] In some embodiments, CARs are constructed with specificity for a particular antigen (or marker or ligand), such as an antigen expressed in a particular cell type to be targeted by adoptive therapy, e.g., a cancer marker, and / or an antigen expressed in a normal or non-diseased cell type, against which an attenuating response is intended to be induced. Thus, a CAR typically comprises, in its extracellular portion, one or more antigen-binding molecules, e.g., one or more antigen-binding fragments, domains, or portions, or one or more antibody-derived variable domains and / or antibody-derived molecules.

[0314] The term "antibody" is used herein in the broadest sense and includes antigen-binding fragments (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (V) fragments capable of specifically binding to an antigen, and H (scFv) regions, single-chain antibody fragments including single-chain variable fragments (sdAb, sdFv, nanobody) and single-domain antibody (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific or trispecific, antibodies, diabodies, triabodies and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term "antibody" should be understood to encompass functional antibody fragments thereof, also referred to herein as "antigen-binding fragments." The term also encompasses intact or whole antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0315] The terms "complementarity-determining region" and "CDR," synonymous with "hypervariable region" or "HVR," are known in the art to refer to non-contiguous sequences of amino acids in an antibody variable region that confer antigen specificity and / or binding affinity. Generally, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. The terms "framework region" and "FR" are known in the art to refer to the non-CDR portions of the heavy and light chain variable regions. Generally, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region.

[0316] The precise amino acid sequence boundaries of a given CDR or FR can be determined using the Kabat numbering scheme (Kabat et al., 1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD; Al-Lazikani et al., (1997) JMB 273,927-948 (Chothia numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," J. Mol. Biol. 262, 732-745 (Contact numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003. Jan;27(1):55-77 (the "IMGT" numbering scheme); Honegger A and Plueckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8;309(3):657-70 (the "Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272 (the "AbM" numbering scheme).

[0317] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. The numbering for both the Kabat and Chothia schemes is based on the length of the most common antibody region sequences, with insertions adjusted by an insertion character, e.g., "30a," and deletions occurring in some antibodies. The two schemes place certain insertions and deletions ("indels") in different positions, resulting in differential numbering. The contact scheme is based on the analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on those used by Oxford Molecular's AbM antibody modeling software.

[0318] Table 1 below lists exemplary boundary positions for CDR-L1, CDR-L2, CDR-L3, and CDR-H1, CDR-H2, and CDR-H3, as identified by the Kabat, Chothia, AbM, and contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are positioned between the CDRs; for example, FR-L1 precedes CDR-L1, FR-L2 is positioned between CDR-L1 and CDR-L2, FR-L3 is positioned between CDR-L2 and CDR-L3, etc. Note that the Kabat numbering scheme shown places insertions at H35A and H35B, so the end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention shown, varies between H32 and H34 depending on the length of the loop. [Table 1]

[0319] Thus, unless otherwise specified, the "CDRs" or "complementary determining regions" of a given antibody or region thereof, e.g., its variable region, or each designated CDR (e.g., CDR-H1, CDR-H2, CDR-H3), will be understood to encompass a (or specific) complementary determining region as defined by any of the schemes described above or other known schemes. For example, a particular CDR (e.g., CDR-H3) may be complementary to a given V H or V L When a variable region amino acid sequence is described as containing the amino acid sequence of a corresponding CDR in the variable region, it is understood that such CDR has the sequence of the corresponding CDR (e.g., CDR-H3) in the variable region as defined by any of the schemes described above or other known schemes. In some embodiments, specific CDR sequences are specified. Although exemplary CDR sequences of the provided antibodies are described using various numbering schemes, it is understood that the provided antibodies can include CDRs described according to any of the other numbering schemes described above or other numbering schemes known to those of skill in the art.

[0320] Similarly, unless otherwise specified, a given antibody or region thereof, e.g., a FR of its variable region or individually designated FR(s) (e.g., FR-H1, FR-H2, FR-H3, FR-H4), will be understood to encompass a (or specific) framework region as defined by any known scheme. In some instances, a scheme for identifying a particular CDR, FR or FRs or CDRs is specified, such as CDRs defined by the Kabat, Chothia, AbM, IMGT, or contact method or other known scheme. In other cases, the specific amino acid sequence of a CDR or FR is given.

[0321] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable regions of the heavy and light chains (V, respectively) of a native antibody H and V L ) generally have a similar structure, with each domain containing four conserved framework regions (FR) and three CDRs (see, e.g., Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)). H or V L Each complementary V domain may be sufficient to confer antigen-binding specificity. L or V H To screen a library of domains, we used V from an antibody that binds to an antigen. H or V L Domains can be used to isolate antibodies that bind to specific antigens. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0322] Among the antigen-binding domains contained in CARs, antigen-binding domains derived from antibody fragments are particularly mentioned. "Antibody fragment" or "antigen-binding fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; heavy chain variable (V); H ) regions, single chain antibody molecules, e.g., scFv, and V H In certain embodiments, the antibody from which the CAR is derived includes, but is not limited to, a heavy chain variable (V) antibody, such as an scFv. H ) region and / or light chain variable (V L ) region.

[0323] A single domain antibody (sdAb) is an antibody fragment that contains all or part of the heavy chain variable region or all or part of the light chain variable region of an antibody. In certain embodiments, a single domain antibody is a human single domain antibody.

[0324] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies or production by recombinant host cells. In some embodiments, the antibody is a recombinantly produced fragment, such as a fragment having two or more antibody regions or chains joined by a synthetic linker, e.g., a peptide linker, and / or a fragment containing a configuration that does not occur in nature, such as a fragment that cannot be produced by enzymatic digestion of a naturally occurring intact antibody. In some aspects, the antibody fragment is an scFv.

[0325] In some embodiments, the CAR is an sdFv, nanobody, V H H and V NAR etc., the variable weight (V) of a monoclonal antibody (mAb) or single domain antibody (sdAb) H ) and variable light (V L In some embodiments, the antigen-binding fragment comprises an antigen-binding portion or portions of an antibody molecule, such as a single-chain antibody fragment (scFv) derived from a variable region of an antibody linked by a flexible linker.

[0326] In some embodiments, the antibody or antigen-binding fragment thereof is a single-chain antibody fragment, such as a single-chain variable fragment (scFv) or a diabody or a single-domain antibody (sdAb). H In some embodiments, the antibody or antigen-binding fragment is a single domain antibody comprising only the heavy chain variable (V H ) region and the light chain variable (V L ) region.

[0327] A "humanized" antibody is an antibody in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A humanized antibody may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of a non-human antibody typically refers to a variant of a non-human antibody that has undergone humanization to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the CDR residues are derived) to, for example, restore or improve antibody specificity or affinity.

[0328] Among the anti-BCMA antibodies from which the provided CARs are derived, human antibodies are particularly preferred. A "human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or a non-human source utilizing a human antibody repertoire, including a human antibody library, or other human antibody coding sequence. This term excludes humanized forms of non-human antibodies containing non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human. This term includes antigen-binding fragments of human antibodies.

[0329] Human antibodies can be prepared by administering immunogens to transgenic animals that have been engineered to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals typically contain all or part of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci or are extrachromosomally present or randomly integrated into the animal's chromosomes. In such transgenic animals, the endogenous immunoglobulin loci are generally inactivated. Human antibodies can also be derived from human antibody libraries, including phage display and cell-free libraries, containing antibody coding sequences derived from the human repertoire.

[0330] Among the antibodies from which the provided CAR is derived, particular mention is made of monoclonal antibodies, including monoclonal antibody fragments. As used herein, the term "monoclonal antibody" refers to an antibody obtained from or contained in a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical except for possible variants that contain naturally occurring mutations or arise during the production of the monoclonal antibody preparation, and such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically contain different antibodies directed against different epitopes, each monoclonal antibody in a monoclonal antibody preparation is directed against a single epitope in an antigen. This term should not be interpreted as requiring the production of antibodies by any particular method. Monoclonal antibodies can be produced by a variety of techniques, including, but not limited to, hybridoma production, recombinant DNA methods, phage display, and other antibody display methods.

[0331] In some embodiments, the CAR comprises the heavy chain variable (V H ) region and / or light chain variable (V L) region, e.g., the BCMA-binding portion(s) of an antibody molecule, such as an scFv antibody fragment. Chimeric receptors such as CARs generally comprise a portion of an antibody molecule, generally the extracellular antigen-binding domain, such as the variable heavy (VH) chain region and / or variable light (VL) chain region of an antibody, e.g., an scFv antibody fragment. In some embodiments, the provided BCMA-binding CARs contain an antibody, such as an anti-BCMA antibody or antigen-binding fragment thereof, that confers the BCMA-binding properties of the provided CAR. In some embodiments, the antibody or antigen-binding domain can be or can be derived from any of the described anti-BCMA antibodies. See, e.g., Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060; U.S. Patent No. 9,034,324, U.S. Patent No. 9,765,342; U.S. Patent Publication Nos. US2016 / 0046724, US20170183418; and International Published PCT Application Nos. WO2016090320, WO2016090327, WO2016094304, WO2016014565, WO106014789, WO2010104949, WO2017 / 025038, or WO2017173256. Any of such anti-BCMA antibodies or antigen-binding fragments can be used in the provided CARs. In some embodiments, the anti-BCMA CARs are variable-weight (V H ) and / or variable light (V L In some embodiments, the antigen-binding domain is an scFv containing a variable heavy (V) region. H ) and / or variable light (V L ) region is derived from an antibody described in WO2016090320 or WO2016090327.

[0332] In some embodiments, the antibody, e.g., anti-BCMA antibody or antigen-binding fragment, comprises a heavy and / or light chain variable (V) as described. H or V LIn some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, contains a V domain sequence, VH1, VH2, and / or VH3, as described. H In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, comprises a V domain sequence containing CDR-L1, CDR-L2, and / or CDR-L3 as described. L In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, comprises a V domain sequence or a sufficient antigen-binding portion thereof containing CDR-H1, CDR-H2, and / or CDR-H3 as described. H V containing the region sequences and containing CDR-L1, CDR-L2 and / or CDR-L3 as described L Particularly included are antibodies having a sequence that is at least or about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to such a sequence.

[0333] In some embodiments, the antibody is a V H V such as any of the sequences (e.g., CDR-H1, CDR-H2, CDR-H3 and / or CDR-H4) H and single domain antibodies (sdAbs) that contain only the domain sequence or a sufficient antigen-binding portion thereof.

[0334] In some embodiments, V H The antibodies (e.g., anti-BCMA antibodies) or antigen-binding fragments thereof provided herein that comprise a V region further comprise a light chain or a sufficient antigen-binding portion thereof. For example, in some embodiments, the antibodies or antigen-binding fragments thereof comprise a V region. H Area and V L Area, or V H and V L In such embodiments, the V H The domain sequence is V HIn some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or scFv. In some such embodiments, the antibody is a full-length antibody that also contains a constant region.

[0335] In some embodiments, the CAR is an anti-BCMA CAR specific for BCMA, for example, human BCMA. Chimeric antigen receptors derived from anti-BCMA antibodies, including mouse anti-human BCMA antibodies and human anti-human BCMA antibodies, and cells expressing such chimeric receptors have been previously described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060, US9,765,342, WO2016 / 090320, WO2016090327, WO2010104949A2, WO2016 / 0046724, WO2016 / 014789, WO2016 / 094304, WO2017 / 025038 and WO2017173256. In some embodiments, the anti-BCMA CAR comprises a variable-heavy (V) antibody derived from an antibody described in WO2016 / 090320 or WO2016090327. H ) and / or variable light (V L In some embodiments, the antigen-binding domain comprises an scFv containing a variable heavy chain (V H ) and variable light chain (V L In some embodiments, the antibody fragment contains a V H The region may be any of the V sequences set forth in any of SEQ ID NOs: 30, 32, 34, 36, 38, 40, 42, 47, 49, 51, 53, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, and 111. H is or comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of the V LThe region may be any of the V sequences set forth in any of SEQ ID NOs: 31, 33, 35, 37, 39, 41, 43, 48, 50, 52, 54, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, and 112. L It is or comprises an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of the region.

[0336] In some embodiments, the antigen binding domain in the anti-BCMA CAR, such as an scFv, is H and V L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 30. H and V L The region is V as set forth in SEQ ID NO: 31. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 32. H and V L The region is V as set forth in SEQ ID NO: 33 L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 34. H and V L The region is V as set forth in SEQ ID NO: 35. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 36 H and V L The region is V as set forth in SEQ ID NO: 37 L In some embodiments, the VH The region is V as set forth in SEQ ID NO: 38 H and V L The region is V as set forth in SEQ ID NO:39 L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 40. H and V L The region is V as set forth in SEQ ID NO: 41. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 42. H and V L The region is V as set forth in SEQ ID NO: 43 L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 77 H and V L The region is V as set forth in SEQ ID NO: 78 L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 79 H and V L The region is V as set forth in SEQ ID NO: 80. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 81. H and V L The region is V as set forth in SEQ ID NO: 82. L In some embodiments, the V HThe region is V as set forth in SEQ ID NO: 83 H and V L The region is V as set forth in SEQ ID NO:84. L In some embodiments, the V H The region is V H and V L The region is V as set forth in SEQ ID NO:86. L In some embodiments, the V H The region is V as set forth in SEQ ID NO:87. H and V L The region is V as set forth in SEQ ID NO:88. L In some embodiments, the V H The region is V as set forth in SEQ ID NO:89. H and V L The region is V as set forth in SEQ ID NO: 90. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 91. H and V L The region is V as set forth in SEQ ID NO: 92. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 93 H and V L The region is V as set forth in SEQ ID NO:94. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 95.H and V L The region is V as set forth in SEQ ID NO:96. L In some embodiments, the V H The region is V H and V L The region is V L In some embodiments, the V H The region is V as set forth in SEQ ID NO:99 H and V L The region is V as set forth in SEQ ID NO: 100. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 101. H and V L The region is V as set forth in SEQ ID NO: 102. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 103. H and V L The region is V as set forth in SEQ ID NO: 104. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 105. H and V L The region is V as set forth in SEQ ID NO: 106. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 107. Hand V L The region is V as set forth in SEQ ID NO: 106. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 30. H and V L The region is V as set forth in SEQ ID NO: 108. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 109. H and V L The region is V as set forth in SEQ ID NO: 110. L In some embodiments, the V H The region is V as set forth in SEQ ID NO: 111. H and V L The region is V as set forth in SEQ ID NO: 112. L The CDR-L1, CDR-L2 and CDR-L3 are contained within the CDR-L1, CDR-L2 and CDR-L3.

[0337] In some embodiments, V H The region is V as set forth in SEQ ID NO:47 H and V L The region is V L In some embodiments, the V H The region is V as set forth in SEQ ID NO:49 H and V L The region is V as set forth in SEQ ID NO: 50. L In some embodiments, the V HThe region is V as set forth in SEQ ID NO: 51. H and V L The region is V as set forth in SEQ ID NO: 52. L In some embodiments, the V H The region is V as set forth in SEQ ID NO:53 H and V L The region is V as set forth in SEQ ID NO:54. L The CDR-L1, CDR-L2 and CDR-L3 are contained within the CDR-L1, CDR-L2 and CDR-L3.

[0338] In some embodiments, the antigen binding domain, such as an scFv, comprises the V set forth in SEQ ID NO: 30. H and V set forth in SEQ ID NO: 31 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 32. H and V set forth in SEQ ID NO: 33 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 34. H and V set forth in SEQ ID NO: 35 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 36. H and V set forth in SEQ ID NO: 37 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 38. H and V set forth in SEQ ID NO: 39 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 40. H and V set forth in SEQ ID NO: 41 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 42. H and V set forth in SEQ ID NO: 43 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 77.H and V set forth in SEQ ID NO: 78 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 79. H and V set forth in SEQ ID NO: 80 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 81. H and V set forth in SEQ ID NO: 82 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 83. H and V set forth in SEQ ID NO: 84 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 85. H and V set forth in SEQ ID NO: 86 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 87. H and V set forth in SEQ ID NO: 88 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 89. H and V set forth in SEQ ID NO: 90 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 91. H and V set forth in SEQ ID NO: 92 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 93. H and V set forth in SEQ ID NO: 94 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 95. H and V set forth in SEQ ID NO: 96 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 97. H and V set forth in SEQ ID NO: 98 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 99. H and V as set forth in SEQ ID NO: 100L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 101. H and V set forth in SEQ ID NO: 102 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 103. H and V as set forth in SEQ ID NO: 104 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 105. H and V set forth in SEQ ID NO: 106 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 107. H and V set forth in SEQ ID NO: 106 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 30. H and V set forth in SEQ ID NO: 108 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 109. H and V set forth in SEQ ID NO: 110 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 111. H and V set forth in SEQ ID NO: 112 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 47. H and V set forth in SEQ ID NO: 48 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO:49. H and V as set forth in SEQ ID NO: 50 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 51. H and V set forth in SEQ ID NO: 52 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 53. H and V set forth in SEQ ID NO: 54 LIn some embodiments, V H or V L is the aforementioned V H or V L In some embodiments, the V has a sequence of amino acids that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of the V sequences and retains binding to BCMA. H The region is V L In some embodiments, the V H The region is V L In some embodiments, the variable heavy chain and the variable light chain are connected by a linker. In some embodiments, the linker is set forth in SEQ ID NO: 55, 70, 72, 73, or 74.

[0339] In some embodiments, the antibody comprises a heavy chain variable (V H ) region and the light chain variable (V L An antibody is an antigen-binding fragment, such as an scFv, that contains one or more linkers joining two antibody domains or regions, such as a V H and V L These include single-chain antibody fragments, such as scFvs and diabodies, particularly human single-chain antibody fragments, which typically include a linker(s) connecting two antibody domains or regions, such as a region. The linker is typically a peptide linker, e.g., a flexible and / or soluble peptide linker, e.g., a linker rich in glycine and serine. Particularly preferred linkers include linkers rich in glycine and serine and / or, in some cases, threonine. In some embodiments, the linker further includes a charged residue, such as lysine and / or glutamic acid, which can improve solubility. In some embodiments, the linker further includes one or more prolines.

[0340] In some aspects, a linker that is rich in glycine and serine (and / or threonine) comprises at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of such amino acid(s). In some embodiments, it comprises at least or about 50%, 55%, 60%, 70%, or 75% glycine, serine, and / or threonine. In some embodiments, the linker is composed substantially entirely of glycine, serine, and / or threonine. Linkers are generally between about 5 and about 50 amino acids in length, typically between at or about 10 and at or about 30, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30, and in some examples, between 10 and 25 amino acids in length. Exemplary linkers include linkers having various numbers of repeats of the sequence GGGGS (4GS; SEQ ID NO: 19) or GGGS (3GS; SEQ ID NO: 71), e.g., between 2, 3, 4, or 5 repeats of such a sequence. Exemplary linkers include linkers having or consisting of the sequence set forth in SEQ ID NO: 72 (GGGGSGGGGSGGGGS), SEQ ID NO: 55 (ASGGGGSGGRASGGGGS), SEQ ID NO: 73 (GSTSGSGKPGSGEGSTKG) or SEQ ID NO: 74 (SRGGGGSGGGGSGGGGSLEMA).

[0341] In some embodiments, the antigen binding domain in the anti-BCMA CAR, such as an scFv, is H and V L In some embodiments, the V H The region includes CDR-H1 set forth in SEQ ID NO: 56, CDR-H2 set forth in SEQ ID NO: 57, and CDR-H3 set forth in SEQ ID NO: 58, and V L The region comprises a CDR-L1 set forth in SEQ ID NO: 59, a CDR-L2 set forth in SEQ ID NO: 60, and a CDR-H3 set forth in SEQ ID NO: 61. HThe region has a sequence of amino acids set forth in SEQ ID NO: 36 or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 36, and L The region has a sequence of amino acids set forth in SEQ ID NO: 37, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 37. In some embodiments, V H The region has the amino acid sequence set forth in SEQ ID NO: 36, L The region has the sequence of amino acids set forth in SEQ ID NO: 37. H The region is V L In some embodiments, the V H The region is V L In some embodiments, the antigen-binding domain is an scFv having a sequence of amino acids set forth in SEQ ID NO: 180, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 180. In certain embodiments, any of the above-described antigen-binding domains binds to BCMA.

[0342] In some embodiments, the antigen binding domain in the anti-BCMA CAR, such as an scFv, is H and V L In some embodiments, the V H The region includes CDR-H1 set forth in SEQ ID NO: 62, CDR-H2 set forth in SEQ ID NO: 63, and CDR-H3 set forth in SEQ ID NO: 64, LThe region comprises CDR-L1 set forth in SEQ ID NO: 65, CDR-L2 set forth in SEQ ID NO: 66, and CDR-H3 set forth in SEQ ID NO: 67. H The region has a sequence of amino acids set forth in SEQ ID NO: 30 or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30, and L The region has a sequence of amino acids set forth in SEQ ID NO: 31, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 31. In some embodiments, V H The region has the amino acid sequence set forth in SEQ ID NO: 30, L The region has the sequence of amino acids set forth in SEQ ID NO: 31. H The region is V L In some embodiments, the V H The region is V L In some embodiments, the antigen-binding domain is an scFv having a sequence of amino acids set forth in SEQ ID NO: 68, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 68. In certain embodiments, any of the above-described antigen-binding domains binds to BCMA.

[0343] In some embodiments, the recombinant receptor, e.g., the antibody portion of a CAR, the recombinant receptor, such as a CAR, comprises a hinge region, e.g., an IgG4 hinge region, an IgG1 hinge region, a C H 1 / C Land / or a spacer, which may be or may include at least a portion of an immunoglobulin constant region or a mutant or modified version thereof, such as an Fc region. In some embodiments, the recombinant receptor further comprises a spacer and / or a hinge region. In some embodiments, the constant region or portion is a constant region or portion of human IgG, e.g., IgG4 or IgG1. In some aspects, the portion of the constant region functions as a spacer region between the antigen recognition component, e.g., the scFv, and the transmembrane domain. The spacer may be of a length that results in increased cellular responsiveness after antigen binding compared to the absence of the spacer.

[0344] Exemplary spacers, e.g., hinge regions, include those described in International Patent Application Publication No. WO2014031687. In some examples, the spacer is 12 amino acids in length, or about 12 amino acids in length, or 12 or fewer amino acids in length. Exemplary spacers have at least about 10-229 amino acids, about 10-200 amino acids, about 10-175 amino acids, about 10-150 amino acids, about 10-125 amino acids, about 10-100 amino acids, about 10-75 amino acids, about 10-50 amino acids, about 10-40 amino acids, about 10-30 amino acids, about 10-20 amino acids, or about 10-15 amino acids, including spacers containing any integer between any of the endpoints of the listed ranges. In some embodiments, the spacer region has about 12 amino acids or fewer, about 119 amino acids or fewer, or about 229 amino acids or fewer. In some embodiments, the spacer is a spacer having at least a particular length, such as having a length that is at least 100 amino acids, such as at least 110, 125, 130, 135, 140, 145, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250 amino acids in length. Exemplary spacers include IgG4 hinge alone, C H 2 and C H IgG4 hinge linked to three domains, or C H Exemplary spacers include an IgG4 hinge alone, C H2 and C H IgG4 hinge linked to three domains, or C H The spacer comprises an IgG4 hinge linked to the 3 domain. Exemplary spacers include, but are not limited to, those described in Hudecek et al., Clin. Cancer Res., 19:3153 (2013), Hudecek et al. (2015) Cancer Immunol Res. 3(2): 125-135, International Patent Application Publication No. WO2014031687, U.S. Patent No. 8,822,647, or Published Application No. US2014 / 0271635. In some embodiments, the spacer comprises an immunoglobulin hinge region, C H 2 and C H In some embodiments, the sequences include three regions: hinge, C H 2 and C H 3 are derived in whole or in part from IgG4 or IgG2. In some cases, the hinge, C H 2 and C H 3 is derived from IgG4. In some embodiments, the hinge, C H 2 and C H One or more of the three are chimeric and contain sequences from IgG4 and IgG2. In some examples, the spacer is an IgG4 / 2 chimeric hinge, an IgG2 / 4 C H 2 and IgG4 C H It contains three regions.

[0345] In some embodiments, the spacer, which may be a constant region or portion thereof of an immunoglobulin, is a spacer of a human IgG, such as IgG4 or IgG1. In some embodiments, the spacer has the sequence ESKYGPPCPPCP (set forth in SEQ ID NO: 1). In some embodiments, the spacer has the sequence set forth in SEQ ID NO: 3. In some embodiments, the spacer has the sequence set forth in SEQ ID NO: 4. In some embodiments, the encoded spacer is or contains the sequence set forth in SEQ ID NO: 29. In some embodiments, the constant region or portion is that of IgD. In some embodiments, the spacer has the sequence set forth in SEQ ID NO: 5. In some embodiments, the spacer has the sequence set forth in SEQ ID NO: 125.

[0346] In some embodiments, the spacer may be derived in whole or in part from IgG4 and / or IgG2 and may contain mutations, such as one or more single amino acid mutations in one or more domains. In some examples, the amino acid modification is a substitution of proline (P) for serine (S) in the hinge region of IgG4. In some embodiments, the amino acid modification is at the C position of the full-length IgG4 Fc sequence. H and substitution of glutamine (Q) for asparagine (N) to reduce glycosylation heterogeneity, such as the N177Q mutation at position 177 in the CH2 region or N176Q at position 176 in the CH2 region of the full-length IgG4 Fc sequence.

[0347] In some embodiments, the spacer can be of a length that provides increased cellular responsiveness following antigen binding compared to the absence of the spacer and / or the presence of a different spacer, e.g., a spacer that differs only in length. In some embodiments, the spacer is at least 100 amino acids in length, e.g., at least 110, 125, 130, 135, 140, 145, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250 amino acids in length. In some examples, the spacer is 12 amino acids or less in length. Exemplary spacers include those having at least about 10-300 amino acids, about 10-200 amino acids, about 50-175 amino acids, about 50-150 amino acids, about 10-125 amino acids, about 50-100 amino acids, about 100-300 amino acids, about 100-250 amino acids, about 125-250 amino acids, or about 200-250 amino acids, including any integer between any of the endpoints of the listed ranges. In some embodiments, the spacer or spacer region is at least about 12 amino acids in length, at least about 119 amino acids or less, at least about 125 amino acids, at least about 200 amino acids, or at least about 220 amino acids, or at least about 225 amino acids in length.

[0348] In some embodiments, the spacer is 125-300 amino acids in length, 125-250 amino acids in length, 125-230 amino acids in length, 125-200 amino acids in length, 125-180 amino acids in length, 125-150 amino acids in length, 150-300 amino acids in length, 150-250 amino acids in length, 150-230 amino acids in length, 150-200 amino acids in length, 150-180 amino acids in length, 180-300 amino acids in length, 180-250 amino acids in length, 180-230 amino acids in length, 180-200 amino acids in length, 200-300 amino acids in length, 200-250 amino acids in length, 200-230 amino acids in length, 230-300 amino acids in length, 230-250 amino acids in length, or 250-300 amino acids in length. In some embodiments, the spacer is at or about at least 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 221, 222, 223, 224, 225, 226, 227, 228, or 229 amino acids in length, or any length between the foregoing.

[0349] Exemplary spacers include spacers conta...

Claims

1. 1. A therapeutic agent comprising a T cell preparation for use in a method for treating multiple myeloma, the method comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM), the T cell preparation comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to BCMA; (b) administering to the subject an immunomodulatory compound, wherein the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione or a pharma- ceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; (c) administering dexamethasone to the subject; A therapeutic agent comprising:

2. 1. A therapeutic agent comprising an immunomodulatory compound for use in a method of treating multiple myeloma, said method comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM) a T cell preparation, the T cell preparation comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA; (b) administering to the subject an immunomodulatory compound, wherein the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione or a pharma- ceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; (c) administering dexamethasone to the subject; A therapeutic agent comprising:

3. 1. A therapeutic agent comprising dexamethasone for use in a method of treating multiple myeloma, said method comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM) a T cell preparation, the T cell preparation comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA; (b) administering to the subject an immunomodulatory compound, wherein the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione or a pharma- ceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof; (c) administering dexamethasone to the subject; A therapeutic agent comprising:

4. 1. A therapeutic agent comprising a T cell preparation, an immunomodulatory compound, and dexamethasone for use in a method of treating multiple myeloma, the method comprising administering the therapeutic agent to a subject having relapsed or refractory multiple myeloma (R / R MM), the T cell preparation comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to BCMA, and the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof.

5. (a) the immunomodulatory compound and dexamethasone are administered simultaneously; and / or (b) the immunomodulatory compound and dexamethasone are administered on the same day; The therapeutic agent according to any one of claims 1 to 3.

6. (a) administration of the immunomodulatory compound is initiated prior to administration of the T cell preparation; and / or (b) administration of the immunomodulatory compound is 0-30 days, 0-15 days, 0-6 days, 0-96 hours, 0-24 hours, 0-12 hours, 0-6 hours, 0-2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours, 6 hours 24 hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days to 30 days, or any number of such dates; The therapeutic agent according to any one of claims 1 to 3.

7. The method of any one of claims 1 to 3, wherein the immunomodulatory compound is administered simultaneously with the administration of the T cell preparation.

8. (a) administration of the immunomodulatory compound is initiated subsequent to administration of the T cell preparation; (b) administration of said immunomodulatory compound begins up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after administration of said T cell preparation; and / or (c) administration of the immunomodulatory compound is initiated at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or about at least 12 days, at least or about at least 14 days, at least or about at least 15 days, at least or about at least 21 days, at least or about at least 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days, or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after administration of the T cell preparation. The therapeutic agent according to any one of claims 1 to 3.

9. The method of any one of claims 1 to 4, wherein the immunomodulatory compound is a pharma- ceutically acceptable salt or polymorph of (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione.

10. The method of any one of claims 1 to 4, wherein the immunomodulatory compound is (S)-3-[4-(4-morpholin-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione.

11. The method according to any one of claims 1 to 4, wherein dexamethasone is administered in a low dose.

12. The method of any one of claims 1 to 4, wherein prior to the method, the subject has not previously been administered the immunomodulatory compound.

13. 1. A therapeutic agent comprising a T cell preparation for use in a method for treating multiple myeloma, the method comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM), the T cell preparation comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to BCMA; (b) administering to the subject a standard triplet regimen; A therapeutic agent comprising:

14. 1. A kit comprising agents for use in a standard triplet regimen for use in a method of treating multiple myeloma, said method comprising: (a) administering to a subject with relapsed or refractory multiple myeloma (R / R MM) a T cell preparation, the T cell preparation comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA; (b) administering to said subject said standard triplet regimen; Including the kit.

15. 1. A kit comprising a T cell preparation and agents for use in a standard triplet regimen for use in a method of treating multiple myeloma, the method comprising administering the T cell preparation to a subject having relapsed or refractory multiple myeloma (R / R MM) and administering the standard triplet regimen to the subject, wherein the T cell preparation comprises a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA.

16. 1. A kit comprising agents for use in a standard triplet regimen for use in a method of treating multiple myeloma, the method comprising administering the standard triplet regimen to a subject with relapsed or refractory multiple myeloma (R / R MM), wherein at the time the standard triplet regimen is initiated, the subject has previously been administered a T cell preparation, the T cell preparation comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that specifically binds to BCMA.

17. The method or kit of any of claims 13 to 16, wherein the standard triplet regimen comprises administration of daratumumab, pomalidomide, and dexamethasone.

18. 18. The treatment or kit of claim 17, wherein prior to the method, the subject has not previously been administered daratumumab in combination with pomalidomide.

19. The method or kit of any of claims 13 to 16, wherein the standard triplet regimen comprises administration of pomalidomide, bortezomib, and dexamethasone.

20. 20. The treatment or kit of claim 19, wherein prior to the method, the subject has not previously been administered pomalidomide in combination with bortezomib.

21. The method or kit of claim 17, wherein dexamethasone is administered in a low dose.

22. 20. The method of claim 19, wherein dexamethasone is administered in a low dose.

23. (a) the standard triplet regimen is initiated prior to administration of the T cell preparation; (b) the standard triplet regimen is administered from 0 to 30 days, 0 to 15 days, 0 to 6 days, 0 to 96 hours, 0 to 24 hours, 0 to 12 hours, 0 to 6 hours, 0 to 2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours, 6 hours to 24 hours, hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 hours to 30 days, 6 hours to 15 days, or 15 days to 30 days before or about any of these; and / or (c) the standard triplet regimen is initiated less than about 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour prior to administration of the T cell preparation; A therapeutic agent or kit according to claim 13 or 14.

24. The method or kit of claim 13 or 14, wherein the standard triplet regimen is administered simultaneously with the administration of the T cell preparation.

25. (a) the standard triplet regimen is initiated following administration of the T cell preparation; (b) the standard triplet regimen is initiated up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days, up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up to about 360 days, or up to or up to about 720 days or more after administration of the T cell preparation; (c) the standard triplet regimen is initiated at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or about at least 12 days, at least or about at least 14 days, at least or about at least 15 days, at least or about at least 21 days, at least or about at least 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days, or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after administration of the T cell preparation; and / or (d) the standard triplet regimen is administered 1 to 360 days, 1 to 240 days, 1 to 180 days, 1 to 120 days, 1 to 90 days, 1 to 60 days, 2 to 360 days, 2 to 240 days, 2 to 180 days, 2 to 120 days, 2 to 90 days, 2 to 60 days, 3 to 360 days, 3 to 240 days, 3 to 180 days, 3 to 120 days, 3 to 90 days, 3 to 60 days, 4 to 360 days, 4 to 240 days, Days later, 4 days to 180 days, 4 days to 120 days, 4 days to 90 days, 4 days to 60 days, 5 days to 360 days, 5 days to 240 days, 5 days to 180 days, 5 days to 120 days, 5 days to 90 days, 5 days to 60 days, 6 days to 360 days, 6 days After 240 days, 6 days after 180 days, 6 days after 120 days, 6 days after 90 days, 6 days after 60 days, 7 days after 360 days, 7 days after 240 days, 7 days after 180 days, 7 days after 120 days, 7 days after 90 days, 7 days after 60 days, 14 days after 36 days 0 days later, 14 days later - 240 days later, 14 days later - 180 days later, 14 days later - 120 days later, 14 days later - 90 days later, 14 days later - 60 days later, 21 days later - 360 days later, 21 days later - 240 days later, 21 days later - 180 days later, 21 days later - 120 days later, 21 days later - 90 days later , 21 days to 60 days, 28 days to 360 days, 28 days to 240 days, 28 days to 180 days, 28 days to 120 days, 28 days to 90 days, 28 days to 60 days, 35 days to 360 days, 35 days to 240 days, 35 days to 180 days, 35 days 17. The therapeutic agent or kit according to any one of claims 13, 14, and 16, wherein the administration is initiated between or about (each including) 49 days to 120 days, 35 days to 90 days, 35 days to 60 days, 42 days to 360 days, 42 days to 240 days, 42 days to 180 days, 42 days to 120 days, 42 days to 90 days, 42 days to 60 days, 49 days to 360 days, 49 days to 240 days, 49 days to 180 days, 49 days to 120 days, 49 days to 90 days, or 49 days to 60 days.

26. 17. The treatment or kit of any of claims 13, 14, and 16, wherein the standard triplet regimen is initiated at or about 56 days (8 weeks) after administration of the T cell preparation, optionally wherein the T cell preparation is administered on day 1 of the method, and the standard triplet regimen is initiated at or about month 3, day 1 (M3D1) of the method.

27. (a) the dose of the engineered T cells is 5×10 ７ ~1 x 10 CAR+ T cells or their approximate number ９ or between 1 x 10 CAR+ T cells or approximately ８ ~1 x 10 CAR+ T cells or their approximate number ９ and / or between or about that number of CAR+ T cells; (b) the dose of the engineered T cells is 4.5×10 ８ or approximately the number of CAR+ T cells; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

28. (a) the method does not include determining a level of soluble BCMA in a tissue sample from the subject; (b) the method does not include determining the level of interleukin-6 (IL-6) in a tissue sample from the subject; and / or (c) the method does not include determining a level of tumor necrosis factor alpha (TNFα) in a tissue sample from the subject. The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

29. (a) the subject has a serum M protein level of about 0.5 g / dL or greater or a urinary M protein level of about 200 mg / 24 hours or greater; or (b) the R / R MM is light chain MM without measurable disease in serum or urine; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

30. 30. The method of claim 29, wherein the R / R MM is light chain MM with no measurable disease in serum or urine and the subject has a serum immunoglobulin free light chain level of about 10 mg / dL or greater or an abnormal serum immunoglobulin kappa lambda free light chain ratio.

31. The method or kit of any of claims 1-4 and 13-16, wherein prior to the method, the subject has undergone one or more prior therapies to treat the R / R MM.

32. (a) the one or more prior therapies includes an immunomodulatory agent; (b) the subject has undergone between 1 and 3 prior therapies to treat the R / R MM, or the subject has undergone at least 3 prior therapies to treat the R / R MM; (c) the one or more prior therapies comprise an anti-CD38 antibody and a proteasome inhibitor; (d) the subject has relapsed or been refractory to said one or more prior therapies; (e) the subject exhibited progressive disease (PD) during or within 6 months of the last of said one or more prior therapies; (f) the subject has had a minimal response or better outcome to at least one of the one or more prior therapies; and / or (g) the one or more prior therapies does not include an allogeneic hematopoietic stem cell transplant, a gene therapy based therapeutic for cancer, an investigational cell therapy for cancer, or a BCMA targeted therapy; The therapeutic agent or kit according to claim 31.

33. (a) the one or more prior therapies included an immunomodulatory agent and the subject has received at least two consecutive cycles of the immunomodulatory agent; and / or (b) the one or more prior therapies comprise an anti-CD38 antibody and a proteasome inhibitor, and the subject has received at least two consecutive cycles of the anti-CD38 antibody and the proteasome inhibitor. The therapeutic agent or kit of claim 32.

34. (a) the T cell preparation is prepared from a leukapheresis product and the subject has not undergone an autologous stem cell transplant (ASCT) within 12 weeks prior to leukapheresis; and / or (b) the T cell preparation is prepared from a leukapheresis product and the subject has not undergone an autologous stem cell transplant (ASCT) within 12 months prior to leukapheresis; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

35. The subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

36. The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16, wherein the CAR comprises an antigen-binding domain that binds to BCMA, a transmembrane domain, and an intracellular signaling region comprising a CD3-zeta (CD3ζ) chain.

37. 37. The method of claim 36, wherein the antigen-binding domain is a single-chain variable fragment (scFv).

38. (a) the antigen-binding domain is Ｈ and V Ｌ region, Ｈ The region comprises CDR-H1 as set forth in SEQ ID NO:62, CDR-H2 as set forth in SEQ ID NO:63, and CDR-H3 as set forth in SEQ ID NO:64, Ｌ The region comprises CDR-L1 set forth in SEQ ID NO:65, CDR-L2 set forth in SEQ ID NO:66, and CDR-H3 set forth in SEQ ID NO:67; (b) the antigen-binding domain comprises: V having a sequence of amino acids set forth in SEQ ID NO:30 or a sequence of amino acids exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:

30. Ｈ A region, V having a sequence of amino acids set forth in SEQ ID NO:31 or a sequence of amino acids exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:

31. Ｌ Area and and / or (c) the antigen-binding domain is an scFv having an amino acid sequence set forth in SEQ ID NO:68, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:68; 37. The therapeutic agent or kit of claim 36.

39. The method or kit of claim 36, wherein the intracellular signaling region further comprises a costimulatory signaling region.

40. (a) the costimulatory signaling region comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof; (b) the costimulatory signaling region comprises the intracellular signaling domain of 4-1BB, optionally human 4-1BB; and / or (c) the costimulatory signaling region is between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain.

40. The therapeutic agent or kit of claim 39.

41. (a) the transmembrane domain is or comprises a transmembrane domain from human CD8; and / or (b) the CAR further comprises an extracellular spacer between the antigen-binding domain and the transmembrane domain.

37. The therapeutic agent or kit of claim 36.

42. (a) the spacer is between 50 or about 50 amino acids and 250 or about 250 amino acids; and / or (b) the spacer is a CD8 hinge; 42. The therapeutic agent or kit of claim 41.

43. (a) the CAR has a sequence as set forth in any one of SEQ ID NOs: 126-177, or a sequence of amino acids that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 126-177; and / or (b) the CAR has an amino acid sequence set forth in SEQ ID NO: 152, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 152; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

44. (a) the T cell preparation comprises idecabutagen viculeucel cells; and / or (b) the T cell preparation is ABECMA®; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.

45. (a) the T cell preparation comprises siltacavtagene autoreucel cells; and / or (b) the T cell preparation is CARVYKTI™; The therapeutic agent or kit according to any one of claims 1 to 4 and 13 to 16.