Combination therapy with T-cell therapy and (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione

Abstract

Provided are methods, compositions, uses, and products for combination therapy, including immunotherapy such as adoptive cell therapy, e.g., T cell therapy, and (S)-3-[4-(4-morpholin-4-ylmethylbenzyloxy)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione, or its enantiomer or enantiomer mixture, or its pharmaceutically acceptable salt, solvate, hydrate, cocrystal, clathrate, or polymorph, for treating subjects with diseases or disorders, such as certain B-cell malignancies, as well as related methods, compositions, uses, and products.The cells generally express a recombinant receptor, such as a chimeric antigen receptor (CAR).In some embodiments, the disease or disorder is non-Hodgkin's lymphoma (NHL), for example, relapsed or refractory NHL, or a specific NHL subtype. TIFF2022554353000025.tif86128

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 932,500, filed on 7 November 2019, entitled "Combination of T-cell therapy with (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione," and U.S. Provisional Patent Application No. 63 / 016,977, filed on 28 April 2020, entitled "Combination of T-cell therapy with (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione," the entire contents of which are incorporated herein by reference.

[0002] Inclusion by referencing sequence listings This application is filed together with an electronic sequence listing. The sequence listing is provided as a file titled 735042022440SeqList.txt, created on November 2, 2020, with a size of 35,330 bytes. The electronic information of the sequence listing is incorporated in its entirety by reference.

[0003] field This disclosure relates, in some aspects, to methods, compositions, uses, and products of combination therapies, including adoptive cell therapy, such as immunotherapy, such as T-cell therapy, for treating subjects having diseases and conditions, such as certain B-cell malignancies, as well as the use of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or its enantiomer or mixture of enantiomers, or its pharmaceutically acceptable salts, solvates, hydrates, cocrystals, inclusion compounds, or polymorphs, as well as related methods, compositions, uses, and products. T-cell therapy includes cells expressing recombinant receptors such as chimeric antigen receptors (CARs). In some aspects, the disease or condition is non-Hodgkin lymphoma (NHL), such as relapsed or refractory NHL or certain NHL subtypes. [Background technology]

[0004] background Various strategies for immunotherapy are available, such as the process of administering engineered T cells for adoptive therapy. For example, strategies are available for engineering T cells that express genetically engineered antigen receptors such as CARs and administering compositions containing such cells to a target. Improved strategies are needed to improve the efficacy of the cells, for example, to improve the persistence, activity, and / or proliferation of the cells upon administration to a target. Methods, compositions, kits, and systems are provided to satisfy such needs. [Overview of the project]

[0005] overview Methods, compositions, uses, and products are provided herein that include combination therapies for subjects having cancer, such as B-cell malignancies, comprising the administration of an immunotherapy, including a cell therapy such as T-cell therapy, and the administration of compound A as described herein. In some aspects, the B-cell malignancy is non-Hodgkin lymphoma (NHL), such as relapsed or refractory NHL or certain NHL subtypes. In some aspects, the methods, uses, and products provided involve the administration of a T-cell therapy, such as CAR-expressing T cells containing an antigen-binding domain that binds to an antigen expressed on B cells. In some aspects, the antigen is CD19.

[0006] (a) A step of administering a T-cell therapy to a subject having a B-cell malignancy, comprising a dose of genetically modified T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19; and (b) Next, the following structure: A step of administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000001.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. Methods for treating B-cell malignancies, including, are provided herein. Here, the administration of the compound is initiated (or has been initiated) within 21 days after the administration of T-cell therapy, and (i) The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. (ii) A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and (iii) A second administration period, which includes a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This will be implemented as part of a cycling regimen.

[0007] In some embodiments, this method has the following structure: The procedure comprises administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000002.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof, wherein the subject has been administered T-cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19 prior to the administration of the compound. The administration of the compound must be initiated (or initiated) within 21 days of the administration of T-cell therapy, and (i) The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. (ii) A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and (iii) A second administration period, which includes a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This will be implemented as part of a cycling regimen.

[0008] In any one aspect of the method provided herein, during each four-week cycle, the compound is not administered for one week after three consecutive weeks in which the compound is administered daily.

[0009] In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg to 0.6 mg or about 0.3 mg to about 0.6 mg during a first administration period.

[0010] In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg to 0.6 mg or about 0.3 mg to about 0.6 mg during the second administration period.

[0011] In some embodiments of any one of the methods provided herein, the second administration period extends for three months, about three months, or more than three months after the start of administration of T-cell therapy. In some embodiments, the second administration period extends for three months, about three months, or more than three months after the start of administration of T-cell therapy. In some embodiments, the second administration period extends up to three months, about three months, or more than three months after the start of administration of T-cell therapy. In some embodiments, the second administration period extends up to three months, or about three months, after the start of administration of T-cell therapy.

[0012] In some embodiments of any one of the methods provided herein, administration of the compound is initiated at or before the peak expansion of T-cell therapy in the subject. In some embodiments, the peak expansion of T-cell therapy occurs between 11 or about 11 days and 15 days or about 15 days after administration of T-cell therapy.

[0013] In some aspect of any one of the methods provided herein, the first administration period begins on the same day as the initiation of T-cell therapy.

[0014] In some embodiments of any one of the methods provided herein, the first administration period begins between 1 day or about 1 day and 15 days or about 15 days after administration of T-cell therapy (including both values). In some embodiments, the first administration period begins between 1 day or about 1 day and 11 days or about 11 days after administration of T-cell therapy (including both values). In some embodiments, the first administration period begins between 8 days or about 8 days and 15 days or about 15 days after administration of T-cell therapy (including both values). In some embodiments, the first administration period begins 1 day or about 1 day after administration of T-cell therapy. In other embodiments of any one of the methods provided herein, the first administration period begins 7 days or about 7 days after administration of T-cell therapy. In other embodiments of any one of the methods provided herein, the first administration period begins 8 days or about 8 days after administration of T-cell therapy. In certain embodiments, the first administration period begins 14 days or approximately 14 days after the administration of T-cell therapy. In certain embodiments, the first administration period begins 15 days or approximately 15 days after the administration of T-cell therapy.

[0015] In some embodiments of any one of the methods provided herein, the rest period begins on day 21 or approximately day 21 after administration of T-cell therapy. In some embodiments, the rest period continues until the target B-cell count level recovers to the same or nearly the same level as measured before the first administration period. In some embodiments, the rest period is one week or approximately one week. In some embodiments, the rest period is approximately one week.

[0016] In some aspects of any one of the methods provided herein, the second administration period begins 28 days after or approximately 28 days after the administration of T-cell therapy.

[0017] In some aspects of any one of the methods provided herein, the second administration period begins 29 days after the administration of T-cell therapy, or approximately 29 days after.

[0018] In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period and / or during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg or about 0.3 mg during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period and during a second administration period.

[0019] In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period and / or during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.45 mg or about 0.45 mg during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period and during a second administration period.

[0020] In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period and / or during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.6 mg or about 0.6 mg during a second administration period. In some aspects of any one of the methods provided herein, the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period and during a second administration period.

[0021] In some embodiments of any one of the methods provided herein, the second administration period ranges from 3 months or about 3 months to 6 months or about 6 months. In some embodiments, the second administration period ranges from about 3 months to about 6 months. In some embodiments, the second administration period ranges from 3 months to 6 months.

[0022] In some embodiments of any one of the methods provided herein, the second administration period extends for three months or approximately three months after the commencement of T-cell therapy. In some embodiments, the second administration period extends up to three months or approximately three months after the commencement of T-cell therapy. In some embodiments, the second administration period extends for at least three months or approximately three months after the commencement of T-cell therapy. In some embodiments, the second administration period extends for at least three months or approximately three months after the commencement of T-cell therapy, and until the subject shows disease progression.

[0023] In some embodiments of any one of the methods provided herein, if a subject achieves a complete response (CR) after treatment more than three months prior to or about three months prior to treatment, or if the cancer, such as a B-cell malignancy, progresses after treatment or relapses after remission, the second administration period extends for three months or about three months after the start of T-cell therapy administration. In some embodiments of any one of the methods provided herein, if a subject achieves a complete response (CR) after treatment more than three months prior to or about three months prior to the start of T-cell therapy administration, or if the B-cell malignancy progresses after treatment or relapses after remission, the second administration period ends at three months or about three months after the start of T-cell therapy administration. In some embodiments, if a subject achieves a complete response (CR) after treatment more than three months prior to or about three months prior to the start of T-cell therapy administration, the second administration period ends at three months or about three months after the start of T-cell therapy administration. In some embodiments, if the B-cell malignancy progresses after treatment or relapses after remission approximately three months or less before the start of T-cell therapy, the second administration period ends three months or approximately three months after the start of T-cell therapy. In some embodiments, if the subject achieves a complete response (CR) within three months, the second administration period extends for three months or approximately three months after the start of T-cell therapy. In some embodiments, if the subject achieves a complete response (CR) within three months, the second administration period ends three months or approximately three months after the start of T-cell therapy.

[0024] In some embodiments of any one of the methods provided herein, the second administration period extends for six months or approximately six months after the commencement of T-cell therapy. In some embodiments, the second administration period extends for up to six months or approximately six months after the commencement of T-cell therapy.

[0025] In some aspects of any one of the methods provided herein, if a subject achieves a complete response (CR) after treatment more than six months prior to or about six months prior to treatment, or if the cancer, such as a B-cell malignancy, progresses after treatment or relapses after remission, the second administration period extends for six months or about six months after the start of T-cell therapy administration. In some aspects of any one of the methods provided herein, if a subject achieves a complete response (CR) after treatment more than six months prior to or about six months prior to the start of T-cell therapy administration, or if the B-cell malignancy progresses after treatment or relapses after remission, the second administration period ends at six months or about six months after the start of T-cell therapy administration. In some aspects of any one of the methods provided herein, if a subject achieves a complete response (CR) after treatment more than six months prior to or about six months prior to the start of T-cell therapy administration, the second administration period ends at six months or about six months after the start of T-cell therapy administration. In some embodiments of any one of the methods provided herein, if the B-cell malignancy progresses after treatment or relapses after remission before or about six months after the initiation of T-cell therapy, the second administration period ends six months or about six months after the initiation of T-cell therapy. In some embodiments, if the subject achieves a complete response (CR) at six months, the second administration period extends for six months or about six months after the initiation of T-cell therapy. In some embodiments, if the subject achieves a complete response (CR) at six months, the second administration period ends six months or about six months after the initiation of T-cell therapy.

[0026] In some embodiments of any one of the methods provided herein, if a subject achieves a partial response (PR) or stable disease (SD) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated six months or about six months after the initiation of T-cell therapy. In some embodiments, if a subject achieves a partial response (PR) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated six months or about six months after the initiation of T-cell therapy. In some embodiments, if a subject achieves stable disease (SD) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated six months or about six months after the initiation of T-cell therapy.

[0027] In some aspect of any one of the methods provided herein, the second administration period ends 12 months or approximately 12 months after the commencement of T-cell therapy administration.

[0028] In some embodiments of any one of the methods provided herein, if a subject achieves a partial response (PR) or stable disease (SD) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated 12 months or about 12 months after the initiation of T-cell therapy. In some embodiments, if a subject achieves a partial response (PR) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated 12 months or about 12 months after the initiation of T-cell therapy. In some embodiments, if a subject achieves stable disease (SD) after treatment prior to or about three months after the initiation of T-cell therapy, the second dose is terminated 12 months or about 12 months after the initiation of T-cell therapy.

[0029] In some aspect of any one of the methods provided herein, if a subject achieves a partial response (PR) or stable disease (SD) after treatment less than three months or about three months after the initiation of T-cell therapy, the second administration period is terminated at least three months or about three months after the initiation of T-cell therapy when the B-cell malignancy progresses or relapses.

[0030] In some aspects of any one of the methods provided herein, the second administration is completed within 12 months or approximately 12 months after the commencement of T-cell therapy.

[0031] In some embodiments of any one of the methods provided herein, the subject achieves a complete response (CR) during the second administration period and before the end of the second administration period. In some embodiments, the second administration period is continued even if the subject achieves a complete response (CR) before the end of the second administration period.

[0032] In some embodiments of any one of the methods provided herein, at the start of administration of the compound, the subject does not exhibit serious toxicity after administration of T-cell therapy. In some embodiments, serious toxicity is severe cytokine release syndrome (CRS), optionally grade 3 or higher, long-term grade 3 or higher, or grade 4 or 5 CRS; and / or serious toxicity is severe neurotoxicity, optionally grade 3 or higher, long-term grade 3 or higher, or grade 4 or 5 neurotoxicity. In some embodiments, serious toxicity is severe CRS. In some embodiments, serious toxicity is grade 3 or higher CRS. In some embodiments, serious toxicity is long-term grade 3 or higher CRS. In some embodiments, serious toxicity is grade 4 CRS. In some embodiments, serious toxicity is grade 5 CRS. In some embodiments, serious toxicity is severe neurotoxicity. In some embodiments, serious toxicity is grade 3 or higher neurotoxicity. In some embodiments, severe toxicity is long-term Grade 3 or higher neurotoxicity. In some embodiments, severe toxicity is Grade 4 neurotoxicity. In some embodiments, severe toxicity is Grade 5 neurotoxicity.

[0033] In some embodiments of any one of the methods provided herein, if a subject exhibits toxicity, optionally hematological toxicity, after administration of the compound, administration of the compound is temporarily suspended and / or the cycling regimen is modified. In some embodiments, if a subject exhibits toxicity after administration of the compound, administration of the compound is temporarily suspended. In some embodiments, if a subject exhibits toxicity after administration of the compound, the cycling regimen is modified. In some embodiments, toxicity is hematological toxicity. In some embodiments, toxicity is selected from severe neutropenia, optionally febrile neutropenia, or prolonged grade 3 or higher neutropenia. In some embodiments, toxicity is febrile neutropenia. In some embodiments, toxicity is prolonged grade 3 or higher neutropenia. In some embodiments, administration of the compound is resumed after the subject no longer exhibits toxicity.

[0034] In some aspects of any one of the methods provided herein, cancer is a B-cell malignancy. In some aspects, B-cell malignancy is lymphoma. In some cases, lymphoma is non-Hodgkin lymphoma (NHL). In some aspects, NHL includes invasive NHL, diffuse large B-cell lymphoma (DLBCL), optionally transformed painless DLBCL-NOS; EBV-positive DLBCL-NOS; T-cell / histiocyte-rich large B-cell lymphoma; primary mediastinal large B-cell lymphoma (PMBCL); follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B); and / or high-grade B-cell lymphoma (double / triple hit) with MYC and BCL2 and / or BCL6 rearrangements with DLBCL histology. In some embodiments, at the time of or immediately before administration of the cell dose, the subject had relapsed after remission or become refractory to therapy after treatment with one, two, or three prior therapies other than one or more prior therapies for lymphoma, optionally NHL, or optionally another dose of CAR-expressing cells. In some embodiments, at the time of or prior to administration of the cell dose, the subject had or had been identified as having double / triple-hit lymphoma; the subject had or had been identified as having chemotherapy-resistant lymphoma, optionally chemotherapy-resistant DLBCL; and / or the subject had not achieved complete remission (CR) in response to prior therapy.

[0035] In any one aspect of the methods provided herein, the subject has been identified or has been identified as having an Eastern Community Oncology Group Performance Status (ECOG) status of less than 1 or 1.

[0036] In some aspects of any of the methods provided herein, the compound is a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of any of the methods provided herein, the compound is a hydrate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of any of the methods provided herein, the compound is a solvate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of any of the methods provided herein, the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same.

[0037] In some aspects of any one of the methods provided herein, the compound is administered orally.

[0038] In some aspects of any one of the methods provided herein, CD19 is human CD19.

[0039] In some embodiments of any one of the methods provided herein, the chimeric antigen receptor (CAR) comprises an extracellular antigen recognition domain that specifically binds to CD19 and an intracellular signaling domain comprising an ITAM. In some embodiments, the intracellular signaling domain comprises a signaling domain of a CD3 zeta (CD3ζ) chain, optionally a human CD3 zeta chain. In some embodiments, the intracellular signaling domain comprises a signaling domain of a human CD3-zeta chain. In some embodiments, the chimeric antigen receptor (CAR) further comprises a co-stimulatory signaling region. In some embodiments, the co-stimulatory signaling region comprises a signaling domain of CD28 or 4-1BB, optionally human CD28 or human 4-1BB. In some embodiments, the co-stimulatory signaling region comprises a signaling domain of human 4-1BB.

[0040] In any one aspect of the methods provided herein, the CAR comprises a CD19-specific scFv; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule, optionally 4-1BB, optionally human 4-1BB, or containing thereof; a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally a CD3 zeta signaling domain, optionally human CD3 zeta signaling domain, or containing thereof; and optionally, the CAR further comprises a spacer between the transmembrane domain and the scFv; the CAR is, in order, CD19-specific. The CAR comprises an scFv; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule, optionally a 4-1BB signaling domain, optionally a human 4-1BB signaling domain, or containing one; a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally a CD3 zeta signaling domain, optionally a human CD3 zeta signaling domain; or the CAR comprises, in order, a CD19-specific scFv; a spacer; a transmembrane domain; and a cytoplasmic signaling domain derived from a co-stimulatory molecule, optionally a 4-1BB signaling domain;Optionally, it includes a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, which is either a CD3 zeta signaling domain or contains one. In some embodiments, the CAR includes a spacer, the spacer being (a) comprising or consisting of all or part of an immunoglobulin hinge or a modified form thereof, or comprising about 15 amino acids or less, and not comprising the CD28 extracellular region or the CD8 extracellular region, (b) comprising or consisting of all or part of an immunoglobulin hinge, optionally an IgG4 hinge, or a modified form thereof, and / or comprising about 15 amino acids or less, and not comprising the CD28 extracellular region or the CD8 extracellular region, or (c) being 12 amino acids long or about 12 amino acids long, and / or comprising all or part of an immunoglobulin hinge, optionally an IgG4 hinge, or a modified form thereof, or (d) the sequences of SEQ ID NO:1, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID (e) A polypeptide spacer comprising or consisting of any variant of the sequence encoded by NO:2, or any variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with such sequence; and / or a cytoplasmic signaling domain derived from a costimulatory molecule comprising SEQ ID NO:12 or any variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with such sequence;and / or cytoplasmic signaling domains derived from primary signaling ITAM-containing molecules include SEQ ID NO:13 or SEQ ID NO:14 or SEQ ID NO:15 having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with them; and / or scFv includes the CDRL1 sequence of RASQDISKYLN (SEQ ID NO:35), the CDRL2 sequence of SRLHSGV (SEQ ID NO:36), and / or the CDRL3 sequence of GNTLPYTFG (SEQ ID NO:37), and / or the CDRH1 sequence of DYGVS (SEQ ID NO:38), the CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO:39), and / or YAMDYWG (SEQ ID The scFv contains the CDRH3 sequence of NO:40), or the scFv contains the variable heavy chain region and variable light chain region of FMC63 and / or the CDRL1 sequence, CDRL2 sequence, CDRL3 sequence, CDRH1 sequence, CDRH2 sequence, and CDRH3 sequence of FMC63, or binds to the same epitope as any of the aforementioned sequences or competes for binding to any of the aforementioned sequences, and optionally the scFv contains, in order, VH, optionally a linker containing SEQ ID NO:41, and VL, and / or the scFv contains a flexible linker and / or the amino acid sequence indicated as SEQ ID NO:42.

[0041] In any one aspect of the methods provided herein, the CAR comprises a CD19-specific scFv; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule, optionally being or containing 4-1BB, optionally human 4-1BB; and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally being or containing a CD3 zeta signaling domain, optionally human CD3 zeta signaling domain; and optionally, the CAR further comprises a spacer between the transmembrane domain and the scFv.

[0042] In any one aspect of the methods provided herein, the CAR comprises, in order: a CD19-specific scFv; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule, optionally a 4-1BB signaling domain, optionally a human 4-1BB signaling domain or containing one; and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally a CD3 zeta signaling domain, optionally a human CD3 zeta signaling domain.

[0043] In any one aspect of the methods provided herein, the CAR comprises, in order, a CD19-specific scFv; a spacer; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule which is optionally a 4-1BB signaling domain; and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule which is optionally a CD3 zeta signaling domain or includes the same.

[0044] In some aspects of any one of the methods provided herein, the spacer is a polypeptide spacer comprising all or part of an immunoglobulin hinge or a modified form thereof, or comprising about 15 amino acids or less. In some aspects of any one of the methods provided herein, the spacer comprises all or part of an immunoglobulin hinge, optionally an IgG4 hinge, or a modified form thereof, and / or comprises about 15 amino acids or less. In some aspects, the spacer is 12 amino acid long or about 12 amino acid long, and / or comprises all or part of an immunoglobulin hinge, optionally IgG4, or a modified form thereof. In some aspect of any one of the methods provided herein, the spacer comprises or consists of the sequence of SEQ ID NO:1, the sequences encoded by SEQ ID NO:2, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, or any variant of said sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with them.

[0045] In any one aspect of the methods provided herein, the cytoplasmic signaling domain derived from the co-stimulatory molecule includes SEQ ID NO:12 or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity.

[0046] In any one aspect of the methods provided herein, cytoplasmic signaling domains derived from primary signaling ITAM-containing molecules include SEQ ID NO:13 or SEQ ID NO:14 or SEQ ID NO:15 having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with them.

[0047] In some aspect of any one of the methods provided herein, scFv includes the CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), the CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and / or the CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), as well as the CDRH1 sequence of DYGVS (SEQ ID NO: 38), the CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and / or the CDRH3 sequence of YAMDYWG (SEQ ID NO: 40). In some aspects of any one of the methods provided herein, scFv comprises the variable heavy chain region and the variable light chain region of FMC63 and / or the CDRL1 sequence, CDRL2 sequence, CDRL3 sequence, CDRH1 sequence, CDRH2 sequence, and CDRH3 sequence of FMC63, and optionally scFv comprises VH containing SEQ ID NO:41 and VL containing the amino acid sequence indicated as SEQ ID NO:42. In some aspects of any one of the methods provided herein, scFv has the amino acid sequence indicated as SEQ ID NO:43.

[0048] In any one aspect of the methods provided herein, the dose of genetically engineered T cells is 1 × 10⁻⁶ 5 ~5×10 8 Or approximately 1 x 10 5 ~5×10 8 Total CAR-expressing T cells, 1 × 10 6 ~2.5×10 8 Or approximately 1 x 106 ~2.5×10 8 of total CAR-expressing T cells, 5×10 6 ~1×10 8 or about 5×10 6 ~1×10 8 of total CAR-expressing T cells, 1×10 7 ~2.5×10 8 or about 1×10 7 ~2.5×10 8 of total CAR-expressing T cells, or 5×10 7 ~1×10 8 or about 5×10 7 ~1×10 8 of total CAR-expressing T cells (including the values at both ends).

[0049] In some embodiments of any one of the methods provided herein, the dose of genetically engineered T cells is at least 1×10 5 or at least about 1×10 5 of CAR-expressing cells, at least 2.5×10 5 or at least about 2.5×10 5 of CAR-expressing cells, at least 5×10 5 or at least about 5×10 5 of CAR-expressing cells, at least 1×10 6 or at least about 1×10 6 of CAR-expressing cells, at least 2.5×10 6 or at least about 2.5×10 6 of CAR-expressing cells, at least 5×10 6 or at least about 5×10 6 of CAR-expressing cells, at least 1×10 7 or at least about 1×10 7 of CAR-expressing cells, at least 2.5×10 7 or at least about 2.5×10 7 of CAR-expressing cells, at least 5×10 7 or at least about 5×10 7 of CAR-expressing cells, at least 1×10 8 or at least about 1×10 8CAR-expressing cells, at least 2.5 × 10⁶ 8 Or at least about 2.5 × 10 8 CAR-expressing cells, or at least 5 × 10⁶ 8 Or at least about 5 x 10 8 Includes CAR-expressing cells.

[0050] In any one aspect of the methods provided herein, the dose of genetically engineered T cells is 5 × 10⁻⁶ 7 Or approximately 5 x 10 7 Includes total CAR-expressing T cells.

[0051] In any one aspect of the methods provided herein, the dose of genetically engineered T cells is 1 × 10⁻⁶ 8 Or approximately 1 x 10 8 Includes CAR-expressing cells.

[0052] In some aspects of any one of the methods provided herein, the dose of cells is administered parenterally, optionally intravenously.

[0053] In some aspect of any one of the methods provided herein, the T cells are primary T cells obtained from a sample derived from the subject.

[0054] In some aspect of any one of the methods provided herein, the T cells are autologous to the subject.

[0055] In some aspect of any one of the methods provided herein, the T cells are allogeneic with respect to the subject.

[0056] In some embodiments of any one of the methods provided herein, the dose of genetically engineered T cells comprises CAR-expressing CD4+ T cells and CAR-expressing CD8+ T cells, and the dose administration comprises the step of administering a plurality of distinct compositions, the plurality of distinct compositions comprising a first composition comprising one of CD4+ T cells and one of CD8+ T cells and a second composition comprising the other of CD4+ T cells or CD8+ T cells. In some embodiments, the first and second compositions are administered at intervals of 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours, or the administration of the first and second compositions is carried out on the same day at intervals of about 0 to about 12 hours, about 0 to about 6 hours, or about 0 to 2 hours, and / or the initiation of the administration of the first composition and the initiation of the administration of the second composition are carried out at intervals of about 1 minute to about 1 hour, or about 5 minutes to about 30 minutes. In some embodiments, the first composition and the second composition are administered at intervals of 2 hours or less, 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, or 5 minutes or less. In some embodiments, the first composition contains CD4+ T cells. In some embodiments, the first composition contains CD8+ T cells. In some embodiments, the first composition is administered before the second composition.

[0057] In some embodiments, the dose comprises a first composition and a second composition, which are administered at intervals of 0 to 12 hours or about 0 to about 12 hours, at intervals of 0 to 6 hours or about 0 to about 6 hours, or at intervals of 0 to 2 hours or about 0 to about 2 hours. In some embodiments, the initiation of administration of the first composition and the initiation of administration of the second composition occur at intervals of 2 hours or about 2 hours, 1 hour or about 1 hour, or 30 minutes or about 30 minutes, 15 minutes or about 15 minutes, 10 minutes or about 10 minutes, or 5 minutes or about 5 minutes. In some embodiments, the initiation and / or completion of administration of the first composition and the completion and / or initiation of administration of the second composition are performed at intervals of 2 hours or less or about 2 hours, 1 hour or less or about 1 hour, or 30 minutes or less or about 30 minutes, at intervals of 15 minutes or less or about 15 minutes, 10 minutes or less or about 10 minutes, or 5 minutes or less or about 5 minutes.

[0058] In some aspects of any one of the methods provided herein, prior to the administration of T-cell therapy, the subject is pre-treated with lymphocyte apheresis including the administration of fludarabine and / or cyclophosphamide. In some aspects of any one of the methods provided herein, the step of administering lymphocyte apheresis including the administration of fludarabine and / or cyclophosphamide to the subject immediately before the administration of T-cell therapy is further included. In some aspects, the lymphocyte apheresis is administered at approximately 200-400 mg / m² 2 Optionally, 300 mg / m² 2 Alternatively, approximately 300 mg / m² 2 Cyclophosphamide (including values ​​at both ends), and / or approximately 20-40 mg / m² 2 Optionally, 30 mg / m² 2 This may include 2-4 days of fludarabine, optionally 3 days of daily administration, or lymphocyte apheresis, at approximately 500 mg / m². 2 This includes the administration of cyclophosphamide. In some embodiments, lymphocyte apheresis therapy is 300 mg / m². 2 Alternatively, approximately 300 mg / m² 2 Cyclophosphamide and approximately 30 mg / m²2 This includes a 3-day daily administration of fludarabine, and / or lymphocyte apheresis therapy at 500 mg / m². 2 Alternatively, approximately 500 mg / m² 2 Cyclophosphamide and approximately 30 mg / m² 2 This includes a 3-day daily dose of fludarabine.

[0059] In some aspects of any one of the methods provided herein, the subject is a human being.

[0060] In any one aspect of the methods provided herein, at least 35%, at least 40%, or at least 50% of subjects treated according to this method achieve a sustained complete response (CR) over 6 months or more than 6 months or 9 months or more, or at least 60, 70, 80, 90, or 95% of subjects achieving CR achieve a sustained complete response (CR); and / or at least 60, 70, 80, 90, or 95% of subjects achieving CR by 6 months remain responsive and maintain CR over 3 months or more than 3 months and / or 6 months or more than 6 months and / or 9 months or more. , and / or survive or survive without progression; and / or at least 50%, at least 60%, or at least 70% of subjects treated according to this method achieve an objective response (OR), and optionally, the OR is persistent for 6 months or more than 6 months or 9 months or more, or persistent in at least 60, 70, 80, 90, or 95% of subjects achieving the OR; and / or at least 60%, 70%, 80%, 90%, or 95% of subjects achieving the OR by 6 months remain responsive or survive for 3 months or more than 3 months and / or 6 months or more.

[0061] In some embodiments of any of the methods provided herein, administration of the compound reverses the depletion phenotype in the subject's CAR-expressing T cells, prevents, inhibits or delays the development of the depletion phenotype in the subject's CAR-expressing T cells, reduces the level or degree of the depletion phenotype in the subject's CAR-expressing T cells, or reduces the proportion of the total number of CAR-expressing T cells in the subject that have the depletion phenotype. In some of any such embodiments, the initiation of the compound administration follows the administration of T cell therapy, and after the administration or initiation of the compound, the subject shows recovery or rescue of antigen-specific or tumor-specific activity or function of the CAR-expressing T cells in the subject, optionally, the recovery, rescue and / or initiation of the compound administration occurs at a point in time after the subject or CAR-expressing T cells in the subject's blood have shown the depletion phenotype.

[0062] In some aspects of the methods provided herein, administration of the compound results in an increase in antigen-specific, e.g., CD19 antigen-specific activity or antigen-receptor-driven activity of naive T cells or non-depleted T cells in a subject, including optionally T cells expressing the CAR, after exposure of T cells to an antigen, e.g., CD19, or an antigen receptor-specific activator, compared to no administration of the compound; or prevents, inhibits, or delays the development of a depletion phenotype in naive T cells or non-depleted T cells in a subject, including optionally T cells expressing the CAR, after exposure of T cells to an antigen, e.g., CD19, or an antigen receptor-specific activator, compared to no administration of the compound; or includes administration in an amount, frequency, and / or duration effective in reversing a depletion phenotype in depleted T cells, including optionally T cells expressing the CAR, in a subject, compared to no administration of the subject. In some of the embodiments, administration of the compound includes administration in an amount, frequency, and duration effective in producing an increase in the activity, preventing, inhibiting, or delaying the onset of the depletion phenotype, and / or reversing the depletion phenotype. In some of the embodiments, administration of the compound includes administration in an amount, frequency, or duration effective in producing an increase in the activity, preventing, inhibiting, or delaying the onset of the depletion phenotype, and / or reversing the depletion phenotype. In some of the embodiments, the T cells in the subject include T cells expressing the CAR, and the antigen is a CD19 target antigen. In some of the embodiments, the T cells in the subject include T cells expressing the CAR, or the antigen is a CD19 antigen.

[0063] In some embodiments of the methods provided herein, the depletion phenotype includes, with respect to T cells or a population of T cells, an increase in the level or degree of surface expression of one or more T cells of one or more depletion markers, optionally two, three, four, five, or six depletion markers, or an increase in the proportion of the population of T cells exhibiting surface expression, compared to a reference T cell population under the same conditions. In some embodiments of the methods provided herein, the depletion phenotype includes, with respect to T cells or a population of T cells, a decrease in the level or degree of activity exhibited by the T cells or population of T cells upon exposure to an antigen or antigen receptor-specific activator, compared to a reference T cell population under the same conditions. In some of any embodiments, the increase in level, degree, or percentage is more than 1.2 times or about 1.2 times, more than 1.5 times or about 1.5 times, more than 2.0 times or about 2.0 times, more than 3 times or about 3 times, more than 4 times or about 4 times, more than 5 times or about 5 times, more than 6 times or about 6 times, more than 7 times or about 7 times, more than 8 times or about 8 times, more than 9 times or about 9 times, more than 10 times or about 10 times, or more. In some of any aspects, the reduction in level, degree, or percentage is greater than 1.2 times or about 1.2 times, greater than 1.5 times or about 1.5 times, greater than 2.0 times or about 2.0 times, greater than 3 times or about 3 times, greater than 4 times or about 4 times, greater than 5 times or about 5 times, greater than 6 times or about 6 times, greater than 7 times or about 7 times, greater than 8 times or about 8 times, greater than 9 times or about 9 times, greater than 10 times or about 10 times, or more.

[0064] In some of the various embodiments, the reference T cell population is a population of T cells known to have a non-depletion phenotype, a population of naive T cells, a population of central memory T cells, or optionally a population of stem central memory T cells that originate from or are of the same species as the subject from which one or more T cells having a depletion phenotype originate. In some of the embodiments of the methods provided herein, the reference T cell population is a target-matched population comprising bulk T cells isolated from the blood of a subject from which one or more T cells having a depletion phenotype originate, optionally the bulk T cells do not express CARs and are obtained from the subject from which one or more T cells having a depletion phenotype originate before receiving a dose of CAR-expressing T cells. In some of the various embodiments, the reference T cell population is a target-matched population comprising bulk T cells isolated from the blood of a subject from which one or more T cells having a depletion phenotype originate, optionally the bulk T cells do not express CARs or are obtained from the subject from which one or more T cells having a depletion phenotype originate before receiving a dose of CAR-expressing T cells. In some of the arbitrary embodiments, the reference T cell population is a composition containing a sample of T cell therapy, or a pharmaceutical composition containing CAR-expressing T cells, prior to its administration to a subject, and optionally, the composition is a cryopreserved sample. In some of the arbitrary embodiments, one or more depletion markers are inhibitory receptors. In some of the arbitrary embodiments, one or more depletion markers are selected from PD-1, CTLA-4, TIM-3, LAG-3, BTLA, 2B4, CD160, CD39, VISTA, and TIGIT.

[0065] In some of the arbitrary embodiments of any of the methods provided herein, the activity is one or more proliferation, cytotoxicity, or production of one or a combination of inflammatory cytokines, and optionally, one or a combination of cytokines is selected from the group consisting of IL-2, IFN-γ, and TNF-α. In some of the arbitrary embodiments, the exposure to the antigen or antigen receptor-specific activator comprises exposing T cells by incubation with the antigen or antigen receptor-specific activator, optionally a CAR-binding activator, wherein the antigen is optionally the CD19 antigen. In some of the arbitrary embodiments, the step of exposing to the antigen or antigen receptor-specific activator comprises the step of incubating T cells with target cells expressing the CD19 antigen, optionally cells of a disease, disorder, or condition such as cancer, e.g., B-cell malignancy.

[0066] In some embodiments, the use of combination therapy comprising T-cell therapy and a compound in a method for treating B-cell malignancies is also provided herein, (a) A step of administering a T-cell therapy to a subject having a B-cell malignancy, comprising a dose of genetically modified T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19; and (b) Next, the following structure: A step of administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000003.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. The compound is administered within 21 days of T-cell therapy and in a cycling regimen comprising: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to 3 consecutive weeks; a rest period of at least 1 week in which the compound is not administered, beginning at the end of the first administration period; and a second administration period comprising a 4-week cycle in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for 3 consecutive weeks in each 4-week cycle.

[0067] In some embodiments, the use of the compound in a method for treating B-cell malignancies is also provided herein, the method having the following structure: The process includes administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000004.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof, wherein the subject is genetically engineered to express a chimeric antigen receptor (CAR) that specifically binds to CD19 prior to the administration of the compound. The patient is receiving T-cell therapy, which includes a dose of T cells, and the compound is administered within 21 days of the administration of T-cell therapy and in a cycling regimen including: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to 3 consecutive weeks; a rest period of at least 1 week in which the compound is not administered, starting from the end of the first administration period; and a second administration period including a 4-week cycle in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for 3 consecutive weeks in each 4-week cycle.

[0068] In some embodiments, the use of combination therapy comprising T-cell therapy and compounds in the manufacture of agents for treating B-cell malignancies is also provided herein, (a) the T-cell therapy to be administered to a subject having a B-cell malignancy, the T-cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19, and (b) the subject subsequently having the following structure: The compound (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000005.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof, should be administered, and administration of the compound should be initiated (or initiated) within 21 days after administration of T-cell therapy and should be carried out in a cycling regimen comprising: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to 3 consecutive weeks; a rest period of at least 1 week in which the compound is not administered, beginning at the end of the first administration period; and a second administration period comprising a 4-week cycle in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for 3 consecutive weeks in each 4-week cycle.

[0069] The use of compounds in the manufacture of drugs for the treatment of B-cell malignancies is also provided herein, and the subject is the following structure: The subject should be administered a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione having TIFF0007837864000006.tif27128, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof, wherein the subject should be given a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19 prior to the administration of the compound. If the patient is receiving T-cell therapy including the compound, administration of the compound should be initiated (or initiated) within 21 days of the administration of the T-cell therapy and should be carried out in a cycling regimen including: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to 3 consecutive weeks; a rest period of at least 1 week in which the compound is not administered, starting from the end of the first administration period; and a second administration period including a 4-week cycle in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for 3 consecutive weeks in each 4-week cycle.

[0070] In some of the uses provided herein, the combination therapy is used in accordance with any of the above embodiments of the methods provided herein.

[0071] In some of the uses provided herein, the compound is used in accordance with any of the above embodiments of the methods provided herein.

[0072] In some embodiments of use provided herein, the compound is administered in an amount of 0.3 mg to 0.6 mg or about 0.3 mg to about 0.6 mg during a first administration period.

[0073] In some embodiments of use provided herein, the compound is administered in an amount of 0.3 mg to 0.6 mg or about 0.3 mg to about 0.6 mg during a second administration period.

[0074] In some aspects of use provided herein, the second administration period extends for three months, approximately three months, or more than three months after the start of T-cell therapy. In some aspects of use provided herein, the second administration period extends for up to three months, or approximately three months, after the start of T-cell therapy.

[0075] In some embodiments of use provided herein, administration of the compound is initiated at or before the peak expansion of T-cell therapy in the subject. In some embodiments, the peak expansion of T-cell therapy occurs between 11 days or about 11 days and 15 days or about 15 days after administration of T-cell therapy.

[0076] In some embodiments of the use provided herein, the first administration period begins on the same day as the initiation of T-cell therapy.

[0077] In some aspects of use provided herein, the first administration period begins between 1 day or about 1 day and 15 days or about 15 days after administration of T-cell therapy (including both extreme values). In some aspects of use provided herein, the first administration period begins between 1 day or about 1 day and 11 days or about 11 days after administration of T-cell therapy (including both extreme values). In some aspects of use provided herein, the first administration period begins between 8 days or about 8 days and 15 days or about 15 days after administration of T-cell therapy (including both extreme values).

[0078] In some embodiments of the use provided herein, the first administration period begins one day or approximately one day after the administration of T-cell therapy. In some embodiments of the use provided herein, the first administration period begins seven days or approximately seven days after the administration of T-cell therapy. In some embodiments of the use provided herein, the first administration period begins eight days or approximately eight days after the administration of T-cell therapy. In some embodiments of the use provided herein, the first administration period begins fourteen days or approximately fourteen days after the administration of T-cell therapy. In some embodiments of the use provided herein, the first administration period begins fifteen days or approximately fifteen days after the administration of T-cell therapy.

[0079] In some embodiments of the use provided herein, the rest period begins on day 21 or approximately day 21 after administration of T-cell therapy. In some embodiments of the use provided herein, the rest period continues until the target B-cell count level recovers to the same or nearly the same level as measured before the first administration period. In some embodiments of the use provided herein, the rest period is approximately one week.

[0080] In some embodiments of the use provided herein, the second administration period begins 28 days or approximately 28 days after the administration of T-cell therapy. In some embodiments of the use provided herein, the second administration period begins 29 days or approximately 29 days after the administration of T-cell therapy.

[0081] In some embodiments of use provided herein, the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period and / or during a second administration period.

[0082] In some embodiments of the use provided herein, the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period and / or during a second administration period.

[0083] In some embodiments of use provided herein, the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period and / or during a second administration period.

[0084] In some aspects of the uses provided herein, the compound is a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of the uses provided herein, the compound is a hydrate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of the uses provided herein, the compound is a solvate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. In some aspects of the use provided herein, the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione or comprises the same.

[0085] In some aspects of use provided herein, B-cell malignancies are lymphomas. In some aspects, lymphoma is non-Hodgkin lymphoma (NHL), and optionally NHL includes invasive NHL, diffuse large B-cell lymphoma (DLBCL), optionally transformed painless DLBCL-NOS; EBV-positive DLBCL-NOS; T-cell / histiocyte-rich large B-cell lymphoma; primary mediastinal large B-cell lymphoma (PMBCL); follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B); and / or high-grade B-cell lymphoma (double / triple hit) with DLBCL histology and MYC and BCL2 and / or BCL6 rearrangements.

[0086] In some aspects of use provided herein, CD19 refers to human CD19.

[0087] In some embodiments of the uses provided herein, a chimeric antigen receptor (CAR) comprises an extracellular antigen recognition domain that specifically binds to CD19 and an intracellular signaling domain comprising an ITAM. In some embodiments, the intracellular signaling domain comprises a CD3 zeta (CD3ζ) chain, optionally a signaling domain of a human CD3 zeta chain.

[0088] In some embodiments of the uses provided herein, the chimeric antigen receptor (CAR) further comprises a co-stimulatory signaling region. In some embodiments, the co-stimulatory signaling region comprises a signaling domain of CD28 or 4-1BB, optionally human CD28 or human 4-1BB. In some embodiments, the co-stimulatory signaling region comprises a signaling domain of human 4-1BB.

[0089] In some embodiments of use provided herein, the dose of genetically engineered T cells is 1 × 10⁻⁶ 5 ~5×10 8 Or approximately 1 x 10 5 ~5×10 8 Total CAR-expressing T cells, 1 × 10 6 ~2.5×10 8 Or approximately 1 x 10 6 ~2.5×10 8 Total CAR-expressing T cells, 5 × 10 6 ~1 × 10 8 Or approximately 5 x 10 6 ~1 × 10 8 Total CAR-expressing T cells, 1 × 10 7 ~2.5×10 8 Or approximately 1 x 10 7 ~2.5×10 8 Total CAR-expressing T cells, or 5 × 10⁶ 7 ~1 × 10 8 Or approximately 5 x 10 7 ~1 × 10 8 This includes total CAR-expressing T cells (including values ​​at both ends). In some aspects of use provided herein, the dose of genetically engineered T cells is at least 1 × 10⁶ 5 Or at least about 1 × 10 5CAR-expressing cells of at least 2.5×10 5 or at least about 2.5×10 5 CAR-expressing cells of at least 5×10 5 or at least about 5×10 5 CAR-expressing cells of at least 1×10 6 or at least about 1×10 6 CAR-expressing cells of at least 2.5×10 6 or at least about 2.5×10 6 CAR-expressing cells of at least 5×10 6 or at least about 5×10[[ID=1**]] 6 CAR-expressing cells of at least 1×10 7 or at least about 1×10 7 CAR-expressing cells of at least 2.5×10 7 or at least about 2.5×10 7 CAR-expressing cells of at least 5×10 7 or at least about 5×10 7 CAR-expressing cells of at least 1×10 8 or at least about 1×10 8 CAR-expressing cells of at least 2.5×10 8 or at least about 2.5×10 8 CAR-expressing cells, or at least 5×10 8 or at least about 5×10 8 include CAR-expressing cells. In some embodiments of the uses provided herein, the dose of genetically engineered T cells is 5×10 7 or about 5×10 7 total CAR-expressing T cells. In some embodiments of the uses provided herein, the dose of genetically engineered T cells is 1×10<******107>or about 1×10 8 total CAR-expressing cells.

[0090] In some aspects of the uses provided herein, the dose of genetically engineered T cells includes CD4+ T cells expressing a CAR and CD8+ T cells expressing a CAR, and administration of the dose includes the step of administering a plurality of separate compositions, the plurality of separate compositions including a first composition containing one of CD4+ T cells and CD8+ T cells and a second composition containing the other of CD4+ T cells or CD8+ T cells. In some aspects, the first composition contains CD4+ T cells. In some aspects, the first composition contains CD8+ T cells. [Invention 1001] A method for treating B-cell malignancies, (a) A step of administering a T-cell therapy to a subject having a B-cell malignancy, comprising a dose of genetically modified T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19; and (b) Next, the following structure: TIFF0007837864000007.tif27128 A step of administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. Includes, The administration of the compound is initiated (or will be initiated) within 21 days after the administration of T-cell therapy, and The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and A second administration period, comprising a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This is implemented in cycling regimens that include method. [Invention 1002] A method for treating B-cell malignancies, The following structure: TIFF0007837864000008.tif27128 The process includes administering to a subject a compound that is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. Prior to the administration of the compound, the subject received T-cell therapy including a dose of genetically modified T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19. The administration of the compound is initiated (or will be initiated) within 21 days after the administration of T-cell therapy, and The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and A second administration period, comprising a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This is implemented in cycling regimens that include method. [Invention 1003] The method of the present invention 1001 or 1002, wherein the compound is administered in an amount of 0.3 mg to about 0.6 mg or about 0.3 mg to about 0.6 mg during a first administration period. <00009५0> [Invention 1004] The method according to any one of the present invention 1001 to 1003, wherein the compound is administered in an amount of 0.3 mg to about 0.6 mg or about 0.3 mg to about 0.6 mg during a second administration period. [Invention 1005] The method according to any one of the present invention 1001 to 1004, wherein the second administration period is 3 months, approximately 3 months, or longer than 3 months after the start of T-cell therapy administration. [Invention 1006] The method according to any one of items 1001 to 1004 of the present invention, wherein the second administration period extends up to three months or approximately three months after the start of T-cell therapy administration. [Invention 1007] Any method 1001 to 1006 of the present invention, wherein the administration of the compound is initiated at or before the peak expansion of T-cell therapy in the subject. [Invention 1008] The method of the present invention 1007, wherein the peak expansion of T-cell therapy occurs between 11 days or about 11 days and 15 days or about 15 days after administration of T-cell therapy. [Invention 1009] The method according to any one of items 1001 to 1008 of the present invention, wherein the first administration period begins on the same day as the start of T-cell therapy administration. [Invention 1010] The method according to any one of the present invention 1001 to 1008, wherein the first administration period is started between 1 day or about 1 day and 15 days or about 15 days (including the values ​​at both ends) after administration of T-cell therapy. [Invention 1011] The method according to any one of the present invention 1001-1008 and 1010, wherein the first administration period begins between 1 day or about 1 day and 11 days or about 11 days (including the values ​​at both ends) after administration of T-cell therapy. [Invention 1012] The method according to any one of the present invention 1001-1008 and 1010, wherein the first administration period is initiated between 8 days or about 8 days and 15 days or about 15 days (including the values ​​at both ends) after administration of T-cell therapy. [Invention 1013] The method according to any one of the present invention 1001-1008, 1010, and 1011, wherein the first administration period begins one day or approximately one day after the administration of T-cell therapy. [Invention 1014] The method according to any of items 1001-1008, 1010, and 1011 of the present invention, wherein the first administration period begins 7 days or approximately 7 days after the administration of T-cell therapy. [Invention 1015] The method according to any of items 1001-1008 and 1010-1012 of the present invention, wherein the first administration period begins 8 days or approximately 8 days after the administration of T-cell therapy. [Invention 1016] The method according to any of items 1001-1008, 1010, and 1012 of the present invention, wherein the first administration period begins 14 days or approximately 14 days after the administration of T-cell therapy. [Invention 1017] The method according to any of items 1001-1008, 1010, and 1012 of the present invention, wherein the first administration period begins 15 days or approximately 15 days after the administration of T-cell therapy. [Invention 1018] The method according to any of items 1001 to 1017 of the present invention, wherein the resting period begins on day 21 or approximately day 21 after administration of T-cell therapy. [Invention 1019] A method according to any one of items 1001 to 1018 of the present invention, wherein a rest period is maintained until the target B cell count level recovers to the same or nearly the same level as measured before the first administration period. [Invention 1020] A method of the present invention, wherein the rest period is approximately one week. [Invention 1021] The method according to any of items 1001 to 1020 of the present invention, wherein the second administration period is started 28 days after or approximately 28 days after the administration of T-cell therapy. [Invention 1022] The method according to any of items 1001 to 1020 of the present invention, wherein the second administration period is started 29 days after or approximately 29 days after the administration of T-cell therapy. [Invention 1023] The method according to any one of items 1001 to 1022 of the present invention, wherein the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period and / or during a second administration period. [Invention 1024] The method according to any one of the invention 1001 to 1022, wherein the compound is administered in an amount of 0.3 mg or about 0.3 mg during a first administration period and during a second administration period. [Invention 1025] The method according to any one of the present invention 1001 to 1022, wherein the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period and / or during a second administration period. [Invention 1026] The method according to any one of items 1001 to 1022 of the present invention, wherein the compound is administered in an amount of 0.45 mg or about 0.45 mg during a first administration period and during a second administration period. [Invention 1027] The method according to any one of the 1001 to 1022 of the present invention, wherein the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period and / or during a second administration period. [Invention 1028] The method according to any one of the present invention 1001 to 1022, wherein the compound is administered in an amount of 0.6 mg or about 0.6 mg during a first administration period and during a second administration period. [Invention 1029] The method according to any one of the 1001 to 1028 of the present invention, wherein the compound is a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. [Invention 1030] The method according to any one of the present invention 1001 to 1028, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione hydrate or contains the same. [Invention 1031] The method according to any one of the present invention 1001 to 1028, wherein the compound is a solvate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. [Invention 1032] The method according to any one of the present invention 1001 to 1028, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or comprises the same. [Invention 1033] The method according to any of items 1001-1004 and 1007-1032 of the present invention, wherein if the subject achieved a complete response (CR) after treatment, or if the B-cell malignancy progressed after treatment or relapsed after remission, more than 3 months or approximately 3 months before the start of T-cell therapy administration, the second administration period is terminated 3 months or approximately 3 months after the start of T-cell therapy administration. [Invention 1034] The method of the present invention 1033, wherein if the subject achieves a complete response (CR) within 3 months, the second administration period is terminated 3 months or approximately 3 months after the start of T-cell therapy administration. [Invention 1035] The method according to any of items 1001-1004 and 1007-1032 of the present invention, wherein the second administration period is terminated 6 months or approximately 6 months after the start of T-cell therapy administration. [Invention 1036] The method according to any of items 1001-1004 and 1007-1032 of the present invention, wherein if the subject achieved a complete response (CR) after treatment, or if the B-cell malignancy progressed after treatment or relapsed after remission, more than 6 months or approximately 6 months before the start of T-cell therapy administration, the second administration period is terminated 6 months or approximately 6 months after the start of T-cell therapy administration. [Invention 1037] The method of the present invention 1036, wherein if the subject achieves a complete response (CR) within 6 months, the second administration period is terminated 6 months or approximately 6 months after the start of T-cell therapy administration. [Invention 1038] A method according to any of items 1001 to 1037 of the present invention, wherein the second administration period is continued even if the subject achieves a complete response (CR) before the end of the second administration period. [Invention 1039] A method according to any one of items 1001 to 1038 of the present invention, wherein the subject does not exhibit serious toxicity after administration of T-cell therapy at the time of initiation of administration of the compound. [Invention 1040] Severe toxicity is severe cytokine release syndrome (CRS), optionally grade 3 or higher, long-term grade 3 or higher, or grade 4 or 5 CRS; and / or Severe toxicity is defined as severe neurotoxicity, optionally grade 3 or higher, long-term grade 3 or higher, or grade 4 or 5 neurotoxicity. The method of the present invention 1039. [Invention 1041] Any method of the present invention 1001 to 1040, wherein if the subject exhibits toxicity, optionally hematological toxicity, after administration of the compound, the administration of the compound is temporarily suspended and / or the cycling regimen is changed. [Invention 1042] The method of the present invention 1041, wherein the toxicity is selected from severe neutropenia, optionally febrile neutropenia, and prolonged grade 3 or higher neutropenia. [Invention 1043] The method of the present invention 1041 or 1042, wherein the administration of the compound is resumed after the subject no longer exhibits toxicity. [Invention 1044] A method according to any of items 1001 to 1043 of the present invention, wherein the B-cell malignant tumor is lymphoma. [Invention 1045] The method of the present invention 1044, wherein the lymphoma is non-Hodgkin lymphoma (NHL), and optionally, the NHL includes invasive NHL, diffuse large B-cell lymphoma (DLBCL), optionally transformed painless DLBCL-NOS; EBV-positive DLBCL-NOS; T-cell / histiocyte-rich large B-cell lymphoma; primary mediastinal large B-cell lymphoma (PMBCL); follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B); and / or high-grade B-cell lymphoma (double / triple hit) with DLBCL histology and MYC and BCL2 and / or BCL6 rearrangements. [Invention 1046] Any method of the present invention 1001 to 1045, wherein CD19 is human CD19. [Invention 1047] A method according to any one of the present invention 1001 to 1046, wherein the chimeric antigen receptor (CAR) comprises an extracellular antigen recognition domain that specifically binds to CD19 and an intracellular signaling domain containing ITAM. [Invention 1048] The method of the present invention 1047, wherein the intracellular signaling domain includes a CD3 zeta (CD3ζ) chain, optionally a human CD3 zeta chain signaling domain. [Invention 1049] The method of the present invention 1047 or 1048, wherein the chimeric antigen receptor (CAR) further comprises a co-stimulatory signaling region. [Invention 1050] The method of the present invention 1049, wherein the co-stimulatory signaling region comprises a signaling domain of CD28 or 4-1BB, optionally human CD28 or human 4-1BB. [Invention 1051] The method of Invention 1049 or Invention 1050, wherein the co-stimulatory signaling region includes the signaling domain of human 4-1BB. [Invention 1052] The CAR comprises a CD19-specific scFv; a transmembrane domain; optionally a cytoplasmic signaling domain derived from a co-stimulatory molecule, which is or contains 4-1BB, optionally human 4-1BB; and optionally a CD3 zeta signaling domain, which is or contains human CD3 zeta signaling domain, derived from a primary signaling ITAM-containing molecule, and Optionally, the CAR further includes a spacer between the transmembrane domain and the scFv. Any method according to invention 1001 to 1051. [Invention 1053] A method according to any one of the present invention 1001 to 1051, wherein the CAR comprises, in order: a CD19-specific scFv; a transmembrane domain; optionally a cytoplasmic signaling domain derived from a co-stimulatory molecule, which is either a 4-1BB signaling domain or a human 4-1BB signaling domain; and optionally a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, which is either a CD3 zeta signaling domain or a human CD3 zeta signaling domain. [Invention 1054] A method according to any one of the present invention 1001 to 1051, wherein the CAR comprises, in order, a CD19-specific scFv; a spacer; a transmembrane domain; a cytoplasmic signaling domain derived from a co-stimulatory molecule which is optionally a 4-1BB signaling domain; and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule which is optionally a CD3 zeta signaling domain or contains the same. [Invention 1055] The spacer, polypeptide spacers comprising all or part of an immunoglobulin hinge or a modified form thereof, or comprising approximately 15 amino acids or less. That is, Any method according to invention 1052 to 1054. [Invention 1056] Any method of the Invention 1052-1055, wherein the spacer comprises or consists of all or part of an immunoglobulin hinge, optionally an IgG4 hinge, or a modified form thereof, and / or comprises about 15 amino acids or less. [Invention 1057] The method of Invention 1055 or Invention 1056, wherein the spacer is 12 amino acid length or approximately 12 amino acid length and / or comprises or consists of all or part of an immunoglobulin hinge, optionally IgG4, or a modified form thereof. [Invention 1058] Any method of the present invention 1052 to 1057, wherein the spacer has or consists of the sequence of SEQ ID NO:1; the sequences encoded by SEQ ID NO:2, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34; or a variant of said sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with them. [Invention 1059] A method according to any one of the present invention 1052 to 1058, wherein the cytoplasmic signaling domain derived from a co-stimulatory molecule includes SEQ ID NO:12 or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity therewith. [Invention 1060] A method according to any one of the invention 1052 to 1059, wherein the cytoplasmic signaling domains derived from primary signaling ITAM-containing molecules include SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:15 having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with them. [Invention 1061] Any method of the present invention 1052 to 1060, wherein scFv includes the CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), the CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and / or the CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), and / or the CDRH1 sequence of DYGVS (SEQ ID NO: 38), the CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and / or the CDRH3 sequence of YAMDYWG (SEQ ID NO: 40). [Invention 1062] Any method of the Invention 1052 to 1061, wherein scFv comprises the variable heavy chain region and the variable light chain region of FMC63 and / or the CDRL1 sequence, CDRL2 sequence, CDRL3 sequence, CDRH1 sequence, CDRH2 sequence, and CDRH3 sequence of FMC63, and optionally scFv comprises VH containing SEQ ID NO:41 and VL containing the amino acid sequence indicated as SEQ ID NO:42. [Invention 1063] The method according to any one of the invention 1052 to 1061, wherein scFv has the amino acid sequence shown in SEQ ID NO:43. [Invention 1064] The dose of genetically modified T cells is 1 × 10 5 ~5×10 8 Or approximately 1 x 10 5 ~5×10 8 Total CAR-expressing T cells, 1 × 10 6 ~2.5×10 8 Or approximately 1 x 10 6 ~2.5×10 8 Total CAR-expressing T cells, 5 × 10 6 ~1×10 8 Or approximately 5 x 10 6 ~1×10 8 Total CAR-expressing T cells, 1 × 10 7 ~2.5×10 8 Or approximately 1 x 10 7 ~2.5×10 8 Total CAR-expressing T cells, or 5 × 10⁶ 7 ~1×10 8 Or approximately 5 x 10 7 ~1×10 8 A method of the present invention, comprising total CAR-expressing T cells (including values ​​at both ends). [Invention 1065] The dose of genetically modified T cells is at least 1 × 10⁶ 5 Or at least about 1 × 10 5 CAR-expressing cells, at least 2.5 × 10⁶ 5 Or at least about 2.5 × 10 5 CAR-expressing cells, at least 5 × 10 5 Or at least about 5 x 105 CAR-expressing cells, at least 1 × 10⁶ 6 Or at least about 1 × 10 6 CAR-expressing cells, at least 2.5 × 10⁶ 6 Or at least about 2.5 × 10 6 CAR-expressing cells, at least 5 × 10 6 Or at least about 5 x 10 6 CAR-expressing cells, at least 1 × 10⁶ 7 Or at least about 1 × 10 7 CAR-expressing cells, at least 2.5 × 10⁶ 7 Or at least about 2.5 × 10 7 CAR-expressing cells, at least 5 × 10 7 Or at least about 5 x 10 7 CAR-expressing cells, at least 1 × 10⁶ 8 Or at least about 1 × 10 8 CAR-expressing cells, at least 2.5 × 10⁶ 8 Or at least about 2.5 × 10 8 CAR-expressing cells, or at least 5 × 10⁶ 8 Or at least about 5 x 10 8 A method of the present invention, comprising CAR-expressing cells, any of items 1001 to 1064. [Invention 1066] The dose of genetically modified T cells is 5 × 10 7 Or approximately 5 x 10 7 A method of the present invention, comprising total CAR-expressing T cells, as described in any of items 1001 to 1065. [Invention 1067] The dose of genetically modified T cells is 1 × 10 8 Or approximately 1 x 10 8 A method according to any of the present invention 1001 to 1065, comprising CAR-expressing cells. [Invention 1068] A method according to any of items 1001 to 1067 of the present invention, wherein the dose of cells is administered parenterally, optionally intravenously. [Invention 1069] The method according to any of the present invention 1001 to 1068, wherein the T cells are primary T cells obtained from the subject. [Invention 1070] A method according to any of the present invention 1001 to 1069, wherein the T cells are of self-derived origin to the target. [Invention 1071] A method according to any of the present invention 1001 to 1068, wherein the T cells are allogeneic to the target. [Invention 1072] A method according to any of the 1001 to 1071 of the present invention, wherein the dose of genetically modified T cells comprises CAR-expressing CD4+ T cells and CAR-expressing CD8+ T cells, and the administration of the dose comprises the administration of a plurality of distinct compositions, the plurality of distinct compositions comprising a first composition comprising one of the CD4+ T cells and the CD8+ T cells, and a second composition comprising the other of the CD4+ T cells or the CD8+ T cells. [Invention 1073] The first composition and the second composition are administered at intervals of 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours, or the administration of the first composition and the second composition are carried out on the same day, at intervals of approximately 0 to approximately 12 hours, approximately 0 to approximately 6 hours, or approximately 0 to 2 hours, and / or The administration of the first composition and the administration of the second composition are performed at intervals of approximately 1 minute to 1 hour, or approximately 5 minutes to 30 minutes. The method of the present invention 1072. [Invention 1074] The method of the present invention 1072 or 1073, wherein the first composition and the second composition are administered at intervals of 2 hours or less, 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, or 5 minutes or less. [Invention 1075] A method according to any one of the present invention 1072 to 1074, wherein the first composition comprises CD4+ T cells. [Invention 1076] A method according to any one of the present invention 1072 to 1074, wherein the first composition comprises CD8+ T cells. [Invention 1077] A method according to any one of the present invention 1072 to 1076, wherein the first composition is administered before the second composition. [Invention 1078] A method according to any one of items 1001 to 1077 of the present invention, wherein, prior to the administration of T-cell therapy, the subject is pre-treated with lymphocyte apheresis including the administration of fludarabine and / or cyclophosphamide. [Invention 1079] Any method of the present invention 1001 to 1077, further comprising the step of administering a lymphocyte apheresis therapy, including the administration of fludarabine and / or cyclophosphamide, immediately before the administration of T-cell therapy. [Invention 1080] Lymphocyte apheresis therapy involves approximately 200-400 mg / m². 2 Optionally, 300 mg / m² 2 Alternatively, approximately 300 mg / m² 2 Cyclophosphamide (including values ​​at both ends), and / or approximately 20-40 mg / m² 2 Optionally, 30 mg / m² 2 This may include daily administration of fludarabine for 2-4 days, or optionally 3 days, or lymphocyte apheresis at approximately 500 mg / m². 2 A method according to Invention 1078 or Invention 1079, comprising the administration of cyclophosphamide. [Invention 1081] Lymphocyte apheresis therapy is administered at 300 mg / m². 2 Alternatively, approximately 300 mg / m² 2 Cyclophosphamide and approximately 30 mg / m² 2 This includes a 3-day daily dose of fludarabine, and / or Lymphocyte apheresis therapy is administered at 500 mg / m². 2 Alternatively, approximately 500 mg / m² 2 Cyclophosphamide and approximately 30 mg / m² 2 This includes a 3-day daily dose of fludarabine. Any method according to invention 1078 to 1080. [Invention 1082] A method according to any of the present invention 1001 to 1081, wherein the subject is a human. [Invention 1083] At least 35%, at least 40%, or at least 50% of subjects treated according to the method described above achieve a complete response (CR) that is sustained or sustained in at least 60, 70, 80, 90, or 95% of subjects achieving a CR over 6 months or more than 6 months or 9 months or more; and / or At least 60, 70, 80, 90, or 95% of subjects achieving CR by 6 months remain responsive, remain CR, and / or survive or survive without progression at 3 months or more than 3 months and / or 6 months or more than 6 months and / or 9 months or more than 9 months; and / or At least 50%, at least 60%, or at least 70% of subjects treated according to the method described above achieve an objective response (OR), and optionally, the OR is sustained for 6 months or more than 6 months or 9 months or more than 9 months, or is sustained in at least 60%, 70%, 80%, 90%, or 95% of subjects achieving the OR; and / or At least 60, 70, 80, 90, or 95% of subjects achieving OR by 6 months remain responsive or survive for 3 months or more and / or 6 months or more. Any method of the present invention 1001 to 1082. [Invention 1084] Any method of the present invention 1045-1083, wherein at the time of administration of a dose of cells or immediately before administration, the subject has relapsed after remission following treatment with one or more prior therapies for NHL, or optionally another dose of CAR-expressing cells, or has become refractory to said therapy. [Invention 1085] At or before administering the dose of cells, The subject has or has been identified as having double / triple-hit lymphoma; The subject has chemotherapy-resistant lymphoma, optionally chemotherapy-resistant DLBCL, or has been identified as having chemotherapy-resistant DLBCL; and / or The subject did not achieve complete remission (CR) in response to previous therapy. Any method of the present invention 1045 to 1084. [Invention 1086] The administration of the aforementioned compound Reverse the depletion phenotype in target CAR-expressing T cells; To prevent, inhibit, or delay the development of the depletion phenotype in target CAR-expressing T cells; To reduce the level or degree of the depletion phenotype in target CAR-expressing T cells; or It reduces the proportion of CAR-expressing T cells in a subject exhibiting a depletion phenotype. Any method according to invention 1001 to 1085. [Invention 1087] Any method of the present invention 1001 to 1086, wherein the administration of the compound is initiated following the administration of T cell therapy, and after the administration or initiation of the compound, the subject shows recovery or rescue of antigen-specific or tumor-specific activity or function of CAR-expressing T cells in the subject, and optionally, the recovery, rescue, and / or initiation of the administration of the compound occurs at a point after the subject or CAR-expressing T cells in the subject's blood have shown a depleted phenotype. [Invention 1088] The administration of the aforementioned compound (a) To result in an increase in antigen-specific activity or antigen-receptor-driven activity of naive T cells or non-depleted T cells in a subject, including optionally T cells expressing the CAR, after exposure of T cells to the CD19 antigen or antigen receptor-specific activator, compared to the absence of administration of the compound; or (b) To prevent, inhibit, or delay the development of a depletion phenotype in naive or non-depleted T cells in a subject, including optionally T cells expressing the CAR, after exposure of T cells to the CD19 antigen or an antigen receptor-specific activator, compared to the absence of administration of the compound; or (c) In order to reverse the depletion phenotype in depleted T cells, including T cells expressing the CAR, in the subject compared to when the subject has not received the aforementioned administration. Any method of the present invention 1001 to 1087, comprising administration in an effective amount, frequency, and / or duration. [Invention 1089] The administration of the aforementioned compound (i) in order to bring about the aforementioned increase in activity, and (ii) To prevent, inhibit, or delay the onset of the depletion phenotype and / or reverse the depletion phenotype A method of the present invention 1088, comprising administration in an effective amount, frequency, and / or duration. [Invention 1090] The method of the present invention 1088 or 1089, wherein the T cells in the subject include T cells expressing the CAR and / or the antigen is CD19. [Invention 1091] Regarding T cells or populations of T cells, the depletion phenotype is, Compared to a reference T cell population under the same conditions, an increase in the level or degree of surface expression of one or more T cells of one or more depletion markers, optionally two, three, four, five, or six depletion markers, or an increase in the proportion of the T cell population showing surface expression; or Compared to a reference T cell population under the same conditions, a decrease in the level or degree of activity exhibited by the T cells or T cell population upon exposure to the CD19 antigen or antigen receptor-specific activator. Any method of the present invention 1086 to 1090, including the above. [Invention 1092] The method of the present invention 1091, wherein the increase in level, degree, or percentage is more than 1.2 times or about 1.2 times, more than 1.5 times or about 1.5 times, more than 2.0 times or about 2.0 times, more than 3 times or about 3 times, more than 4 times or about 4 times, more than 5 times or about 5 times, more than 6 times or about 6 times, more than 7 times or about 7 times, more than 8 times or about 8 times, more than 9 times or about 9 times, more than 10 times or about 10 times, or more. [Invention 1093] The method of the present invention 1091, wherein the reduction in level, degree, or percentage is greater than 1.2 times or about 1.2 times, greater than 1.5 times or about 1.5 times, greater than 2.0 times or about 2.0 times, greater than 3 times or about 3 times, greater than 4 times or about 4 times, greater than 5 times or about 5 times, greater than 6 times or about 6 times, greater than 7 times or about 7 times, greater than 8 times or about 8 times, greater than 9 times or about 9 times, greater than 10 times or about 10 times, or more. [Invention 1094] The method according to any one of the invention 1091 to 1093, wherein the reference T cell population is a population of T cells known to have a non-depletion phenotype, a population of naive T cells, a population of central memory T cells, or a population of stem central memory T cells that originate from or are of the same species as the subject from which one or more T cells having a depleted phenotype originate. [Invention 1095] (a) The reference T cell population is a target-matched population that includes bulk T cells isolated from the subject's blood from which one or more T cells with a depletion phenotype originate, and optionally the bulk T cells do not express CAR; and / or (b) A reference T cell population obtained from a subject from which one or more T cells with a depleted phenotype are derived, prior to receiving a dose of CAR-expressing T cells. Any method of the present invention 1091 to 1094. [Invention 1096] The method according to any of items 1091 to 1095 of the present invention, wherein the reference T cell population is a composition containing a T cell therapy sample prior to its administration to a subject, or a pharmaceutical composition containing CAR-expressing T cells, and optionally the composition is a cryopreserved sample. [Invention 1097] Any method according to invention 1091 to 1096, wherein one or more depletion markers are inhibitory receptors. [Invention 1098] A method according to any one of the present invention 1091 to 1097, wherein one or more depletion markers are selected from PD-1, CTLA-4, TIM-3, LAG-3, BTLA, 2B4, CD160, CD39, VISTA, and TIGIT. [Invention 1099] Any method according to items 1091 to 1098 of the present invention, wherein the activity is one or more proliferation, cytotoxicity, or production of one or a combination of inflammatory cytokines, and optionally one or a combination of cytokines is selected from the group consisting of IL-2, IFN-γ, and TNF-α. [Invention 1100] Any method 1091 to 1099 of the present invention, wherein exposure to a CD19 antigen or antigen receptor-specific activator comprises incubation with a CD19 antigen or antigen receptor-specific activator, optionally an activator that binds to the antigen-binding domain of a CAR. [Invention 1101] The method of the present invention 1100, comprising exposure to a CD19 antigen or an antigen receptor-specific activator to T cells, target cells expressing the CD19 antigen, and optionally B-cell malignant tumor cells. [Brief explanation of the drawing]

[0091] [Figure 1] Figure 1 shows the intracellular expression of Ikaros and Aiolos in CAR-expressing T cells stimulated with anti-CD19 after incubation with various concentrations of compound A (square) or compound B (circle). [Figure 2A] Figure 2A shows cytokine products from newly thawed or chronically stimulated (e.g., exhibiting a depletion phenotype) anti-CD19 CAR T cells cultured with K562.CD19 target cells for 5 days. [Figure 2B] Figure 2B shows the cytolytic activity of newly thawed and chronically unstimulated (e.g., exhibiting a naive phenotype) or chronically stimulated by 5 days of culture with K562.CD19 target cells (e.g., exhibiting a depletion phenotype). [Figure 2C] Figure 2C shows the proliferation (mean ± SEM) of anti-CD19 CAR T cells from three donors in the presence of compound A (triangles) or compound B (circles) at various concentrations. [Figure 2D] Figure 2D shows the effect of various concentrations of compound A (squares) or compound B (circles) on cell viability when CAR T cells are stimulated with 3 μg / mL anti-ID (left panel) or 30 μg / mL anti-ID (right panel). [Figure 2E] Figure 2E shows the cell cycle analysis of anti-CD19 CAR T cells after treatment with 1000 nM compound B or 100 nM compound A. [Figure 2F] Figure 2F shows the percentage of anti-CD19 CAR T cells in the G1 phase of the cell cycle after exposure to various concentrations of compound A (squares) or compound B (circles). CAR T cells were exposed to 3 μg / mL anti-ID (left panel) or 30 μg / mL anti-ID (right panel). [Figure 2G] Figure 2G shows the intracellular cytokine expression levels of IFNγ, perforin, granzyme B, and IL-2 in anti-CD19 CAR T cells stimulated with 30 μg / mL anti-ID for 24 hours (left panel) or 72 hours (right panel) and exposed to compound A or compound B. [Figure 3A] Figure 3A shows the expression of Ikaros in anti-CD19 CAR T cells subjected to chronic stimulation in the presence of compound A (10 nM or 100 nM). [Figure 3B] Figure 3B shows the cell lysis activity, measured by the number of tumor cells over time, for chronically stimulated cells co-incubated in the presence of compound A (0.001 μM or 0.01 μM) compared to the absence of the compound (control), before rechallenge by target cells expressing CD19. [Figure 3C] Figure 3C shows the size of Granta-519 tumor spheroids at various time points after co-culture with 1 μM compound B (left panel) or 0.001 μM or 0.01 μM compound A (right panel). [Figure 3D]Figure 3D shows the mean tumor volume of Granta-519 tumor spheroids 9 days after compound A was discovered. [Figure 3E] Figure 3E shows a representative image of Granta-519 tumor cells grown as three-dimensional spheroids on day 9 after co-culture with anti-CD19 CAR T cells following co-incubation with chronic stimulation and compound A. [Figure 3F] Figure 3F shows cytokine levels of IFNγ, IL-2, and TNFα measured from the supernatant of chronically stimulated anti-CD19 CAR T cells co-cultured with CD19 tumor spheroids for 5 days and treated with compound A or compound B. [Figure 3G] Figure 3G shows the mean IFNγ concentrations from the supernatant measured after 5 days of co-culture, determined from pooled data from three donors and two independent experiments (statistically significant differences between treatments are indicated as *P<.05, ***P<.001, and ****P<.0001). [Figure 3H] Figure 3H shows a volcano plot illustrating differentially expressed genes induced by each simultaneous treatment with 1 nM or 10 nM of compound A during chronic stimulation. [Figure 3I] Figure 3I shows a comparison of the effects of chronic stimulation and compound A 10nM during chronic stimulation on gene expression profiles (log2 ratio changes). [Figure 3J] Figure 3J shows the KEGG pathway enrichment analysis of differentially expressed genes. [Figure 4A] Figure 4A shows the effect of compound A (0.001 μM or 0.01 μM) on the cytolytic activity of anti-CD19 CAR T cells targeting K562 cells (K562.CD19), Raji cells, or Granta-519 cells transduced with CD19. [Figure 4B] Figure 4B shows the mean size measurements of Granta-519 tumor spheroids 9 days after co-culture with CAR T cells. [Figure 4C]Figure 4C shows cytokine levels of IFNγ, IL-2, and TNFα measured from the supernatant of chronically stimulated anti-CD19 CAR T cells co-cultured with CD19 tumor spheroids for 5 days and treated with compound A or compound B. [Figure 5A] Figure 5A shows the mean measured size of A549.CD19 tumor spheroids on day 9 after co-culture with CAR-T cells in the presence of compound A (0.001 μM, 0.01 μM, or 0.1 μM). [Figure 5B] Figure 5B shows the multiplier change in the number of CAR T cells in co-cultures with A549.CD19 tumor spheroids, measured on day 5 after treatment with compound A (0.01 μM or 0.1 μM). [Figure 5C] Figure 5C shows representative images of Granta-519 spheroids and A549.CD19 spheroids after 9 days of co-culture with chronically stimulated anti-CD19 CAR T cells and rescue incubation with compound A. [Figure 5D] Figure 5D shows the mean measured size of tumor spheroids over time after co-culture of anti-CD19 CAR T cells with compound A. [Figure 5E] Figure 5E shows the cytokine levels of IFNγ, IL-2, and TNFα measured from the supernatant of chronically stimulated anti-CD19 CAR T cells co-cultured with CD19 tumor spheroids for 5 days and treated with compound A. [Figure 5F] Figure 5F shows the mean IFNγ concentrations measured from the co-culture supernatant on day 5, pooled from data from three donors and two independent experiments (statistically significant differences between treatments are indicated as *P<.05 and ****P<.0001). [Figure 5G] Figure 5G shows a volcano plot illustrating differentially expressed genes induced by 9 days of co-culture with chronically stimulated anti-CD19 CAR T cells, followed by rescue treatment with compound A. [Figure 5H]Figure 5H shows a comparison of the effects on gene expression profiles (log2 ratio changes) induced by rescue treatment with 10 nM compound A on chronically stimulated anti-CD19 CAR T cells. [Figure 5I] Figure 5I shows KEGG pathway enrichment analysis of differentially expressed genes in chronically stimulated anti-CD19 CAR T cells after rescue treatment with compound A. [Figure 6A] Figure 6A shows the cytolytic activity of anti-CD19 CAR T cells co-cultured with RL CD19+ tumor cells in the presence of compound A (0.001 μM, 0.01 μM, or 0.1 μM), as measured by the number of tumor cells. [Figure 6B] Figure 6B shows the tumor size of RL tumor spheroids co-cultured with anti-CD19 CAR T cells in the presence of compound B (1 μM) or compound A (0.001 μM, 0.01 μM, or 0.1 μM). [Figure 6C] Figure 6C shows the number of tumor cells in RL tumor spheroids co-cultured with anti-CD19 CAR T cells in the presence of compound B (1 μM) or compound A (0.001 μM, 0.01 μM, or 0.1 μM). [Figure 7A] Figure 7A is a heatmap of intracellular cytokines after 3 days of treatment of anti-CD19 CAR T cells with compound A. The figure shows the log2 factor change in mean fluorescence intensity (MFI) of cytokines compared to the vehicle control in culture. [Figure 7B] Figure 7B shows the cytolytic activity, based on the number of tumor cells, after co-culturing anti-CD19 CAR T cells treated with compound A for 3 days with Raji or Granta-519 lymphoma target cells. [Figure 7C] Figure 7C shows the proliferation of anti-CD19 CAR T cells after treatment with compound A for 3 days. [Figure 7D] Figure 7D shows a plot of the percentage of anti-CD19 CAR T cells in the G1 phase after treatment with compound A for 3 days (mean ± SEM from pooled data from 3 donors and 2 independent experiments). [Modes for carrying out the invention]

[0092] Detailed explanation Manipulated cells such as T cells (e.g., CAR-T cells) for the treatment of subjects with cancer or proliferative disorders, as well as (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione TIFF0007837864000009.tif28128 or pharmaceutically acceptable salts, solvates, hydrates, stereoisomers, tautomers or racemic mixtures thereof (compound A), and methods and uses of their compositions are provided. In some aspects, T cell therapy is adoptive T cell therapy comprising T cells that specifically recognize and / or target antigens associated with cancer or proliferative disorders, e.g., antigens associated with B-cell malignancies, e.g., non-Hodgkin lymphoma or its subtypes. In some aspects, T cell therapy comprises T cells engineered with chimeric antigen receptors (CARs) that bind to antigens, e.g., antigen-binding domains that specifically bind. In some cases, the antigen targeted by T cell therapy is CD19. Also provided are products such as compositions comprising T cell therapy and / or combinations and kits comprising compositions comprising compound A, as well as the use of such compositions and combinations for treating or preventing diseases, conditions, and disorders, including cancers such as B-cell malignancies.

[0093] (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione (compound A) is a cereblon E3 ligase modulating compound (CELMoD). Compound A modulates CRBN, induces ubiquitination of transcription factors Aiolos and Ikaros, increases their proteasome-dependent degradation, and enhances T cell function. Compound A binds more strongly to CRBN and is more efficient at degrading Aiolos and Ikaros than lenalidomide and pomalidomide, and has a potent direct antiproliferative effect against lymphoma cells. Compound A is also 10 to 20 times more potent in the degradation of Ikaros and Aiolos compared to compound B. Compound A has a direct antiproliferative effect against lymphoma cells. As shown herein, compound A also enhances T cell function.

[0094] T cell-based therapies, such as adoptive T cell therapy (including the administration of cells expressing chimeric receptors specific to the disease or disorder of interest, such as chimeric antigen receptors (CARs) and / or other recombinant antigen receptors, as well as other adoptive immunotherapy and adoptive T cell therapies), may be effective in treating diseases and disorders such as B cell malignancies. The manipulated expression of recombinant receptors, such as chimeric antigen receptors (CARs), on the surface of T cells can alter the direction of T cell specificity. In clinical trials, CAR-T cells, such as anti-CD19 CAR-T cells, have produced sustained complete responses in patients with both leukemia and lymphoma (Porter et al. (2015) Sci Transl Med 7:303ra139; Kochenderfer (2015) J. Clin. Oncol., 33:540-9; Lee et al. (2015) Lancet, 385:517-28; Maude et al. (2014) N Engl J Med, 371:1507-17).

[0095] In certain situations, the available approaches to adoptive cell therapy may not always be completely satisfactory. For example, while persistence of CAR T cells can be detected in many subjects with lymphoma, complete responses (CRs) are observed less frequently in subjects with NHL compared to those with ALL. More specifically, higher overall response rates (CR rates of 47%–60%) of up to 80% have been reported after CAR T cell infusion, but in some cases the response is transient, and subjects have been shown to relapse in the presence of persistent CAR T cells (Neelapu, 58th Annual Meeting of the American Society of Hematology (ASH):2016;San Diego, CA, USA. Abstract No. LBA-6.2016; Abramson, Blood. 2016 Dec 01;128(22):4192). Another study reported a long-term CR rate of 40% (Schuster, Ann Hematol. 2016 Oct;95(11):1805-10).

[0096] In some aspects, the explanation for this is immunological depletion of circulating CAR-expressing T cells and / or alterations in the T lymphocyte population. This is because, in some situations, the optimal effect may depend on the ability of administered cells to become active, expand, exhibit various effector functions including cytotoxic death and secretion of various factors such as cytokines, survive including long-term, differentiate into specific phenotypic states (such as long-lived memory state, poorly differentiated state, and effector state), participate in transition or reprogramming, avoid or reduce immunosuppressive states in the local microenvironment of the disease, provide effective and robust recall responses after clearance and re-exposure to target ligands or antigens, and avoid or reduce differentiation into depletion, anergy, peripheral immune tolerance, terminal differentiation, and / or suppressive states.

[0097] In some embodiments, the exposure, persistence, and function of the manipulated cells decrease or decline after administration to the subject. Nevertheless, in some cases, it has been observed that administered cells expressing recombinant receptors can re-expand and / or reactivate in vivo (e.g., showing an increase in cell number or an extension of duration), improving efficacy and therapeutic outcomes in adoptive cell therapy.

[0098] In some aspects, after prolonged stimulation or exposure to antigens and / or exposure under conditions in the tumor microenvironment, T cells may become hypofunctional over time and / or exhibit characteristics associated with depletion. In some aspects, this reduces the persistence and efficacy of T cells against antigens, limiting their ability to be effective. There is a need for methods to improve the efficacy and function of CAR T cells, particularly to minimize, reduce, prevent, or reverse hypofunctional or depleted states.

[0099] The methods provided are based on the observation that certain immunomodulatory compounds, e.g., compound A, improve T cell function, including the ability of T cells to produce one or more cytokines, and functions related to cytotoxicity, expansion, proliferation, and persistence. In some aspects, the methods provided enhance or modulate the proliferation and / or activity of T cell activity associated with the administration of T cell therapy (e.g., CAR-expressing T cells). Such methods and uses have been found to provide or achieve improved or greater T cell functionality, thereby improving antitumor effects.

[0100] In addition to enhancing T cell function, it is also found herein that such immunomodulatory compounds, e.g., compound A, may exhibit effects that reverse, delay, or prevent T cell depletion, including increasing T cell signaling and / or altering one or more genes that are differentially regulated after chronic stimulation. Thus, in some cases, agents that increase or enhance T cell activity may lead cells to a depleted state, but it is found herein that the activity of such immunomodulatory compounds, e.g., compound A, to exert an enhancing effect on T cell activity is cleaved from T cell depletion. Furthermore, observations herein indicate that immunomodulatory compounds, e.g., compound A, exhibit activity that rescues T cells from T cell depletion, such as by restoring or partially restoring the activity of one or more T cells after the cells have exhibited depletion characteristics. Notably, the results herein indicate that exposure of chronically stimulated T cells exhibiting depleted T cell characteristics to the immunomodulatory compounds described herein, e.g., compound A, can restore or partially restore their activity. The observations herein support the possibility that the methods provided may also achieve improved or more sustained responses compared to certain alternative methods, for example, in specific groups of subjects being treated.

[0101] The observations herein demonstrate improved T cell functionality after treatment with compound A on acutely stimulated anti-CD19 CAR T cells and anti-CD19 CAR T cells that were made hypofunctional in chronic stimulation assays. Treatment with compound A was shown to completely degrade the expression of both Ikaros and Aiolos in activated anti-CD19 CAR T cells after 24 hours of treatment. Compound A was shown to increase effector cytokine production (e.g., IFN-γ) in anti-CD19 CAR T cells while simultaneously slowing their proliferation rate. This effect on proliferation was observed at all concentrations tested (1–100 nM) and was attributed to the accumulation of G1 phase anti-CD19 CAR T cells. This demeritation of effector cytokine production from proliferation rate may be clinically beneficial. Concurrent chronic stimulation and treatment with compound A (1 and 10 nM) were shown to limit the occurrence of hypofunctional depletion of anti-CD19 CAR T cells and improve effector cytokine secretion and expression, as assessed by cytolysis against CD19+ lymphoma cell lines and CD19+ lymphoma spheroids. Addition of compound A (1 and 10 nM) to depleted anti-CD19 CAR T cells restored cytolytic activity against CD19+ spheroids and improved effector cytokine expression. In summary, the combination of compound A and anti-CD19 CAR T cells may offer a useful therapeutic approach to enhance and prolong anti-CD19 CAR T cell activity across B-cell malignancies by modulating the tumor microenvironment, improving the sustained antitumor function of CAR T cells, and potentially by a direct antitumor effect against lymphoma cells (Lonial, 2019 J Clin Oncol., 37:8006).

[0102] These observations were made using a chronic stimulation assay to induce hypofunctionality in CAR T cells (e.g., reduced cytolysis and IL-2 secretion). Using this model, CAR T cells were examined to evaluate the effect of compound A on CAR T cell function when present during (concurrent) or after (rescue) exposure to conditions that result in a hypofunctional depletion state. Upon rechallenge with the antigen, the findings provided herein demonstrate that concurrent treatment of CAR T cells during such conditions reverses activity and phenotype, including genetic signatures, associated with CAR T cell hypofunctionality, and preserves more effector function. Similarly, the results indicate that compound A can rescue or restore T cell function, including cytokine production and cytolytic activity, in depleted T cells.

[0103] The observations provided herein also demonstrate that compound A increases effector cytokine production by CAR T cells while simultaneously slowing their proliferation rate. This result is not due to the compound's effect on T cell viability. This effect on proliferation was observed at various concentrations and was attributed to the accumulation of G1 phase T cells. This decoupling of effector cytokine production from proliferation rate may be clinically beneficial, for example, by limiting T cell differentiation in vivo, which can limit efficacy.

[0104] The provided method demonstrates that compound A improves T cell function in engineered T cell therapies, including functions related to T cell expansion, proliferation, growth, and persistence. In some embodiments, the method is advantageous by administering T cell therapies, such as compositions containing cells for adoptive cell therapy, such as T cell therapy (e.g., CAR-expressing T cells), in combination with compound A. In some embodiments, the provided method and use provide or achieve an improved or more persistent response or effect compared to certain alternative methods. In some aspects, the provided method enhances or modulates the proliferation and / or activity of T cell activity associated with the administration of T cell therapy (e.g., CAR-expressing T cells). In certain embodiments, combination therapy with compound A may provide a useful therapeutic approach to enhance and prolong CAR T cell activity across B cell malignancies by modulating the tumor microenvironment and improving the sustained antitumor function of CAR T cells. In some cases, the compound may also have a direct antitumor effect against lymphoma cells.

[0105] Compound A is a multifaceted small molecule immunomodulator that can directly impair primary tumor growth, modulate the immunosuppressive tumor microenvironment, and promote a more robust anti-tumor inflammatory response. Compound A exerts antiproliferative activity against B cells and is being evaluated as a monotherapy targeting B-cell lymphoid malignancies. Compound A and other immunomodulators such as lenalidomide have been shown to directly affect malignant lymphocyte survival through the degradation of Ikaros family transcription factors. The molecular target of Compound A has been identified as the protein cereblon (CRBN), a substrate receptor for the Cullin 4 RING E3 ubiquitin ligase complex. Binding of Compound A to the hydrophobic tri-tryptophan pocket within CRBN promotes the recruitment, ubiquitination, and subsequent proteasomal degradation of several protein substrates, including Aiolos (IKZF3) and Ikaros (IKZF1).

[0106] In the studies described herein, 1 nM and 10 nM concentrations of compound A delayed the onset of CD19 CAR T cell depletion and rescued anti-CD19 CAR T cells from depletion. Doses of 0.3 mg and 1.0 mg of compound A in healthy subjects showed Cmaxes of 2.41 nM to 11.79 nM. In some cases, the dose is an effective dose for mediating the immunomodulatory effect of the compound. In some cases, the dose is a dose that is not expected to cause serious toxicity such as grade 3 neutropenia and dermatitis. In some cases, the methods provided minimize or avoid toxicity after administration to T cell therapy and / or compound A subjects. In some cases, the methods provided herein involve administering a dose substantially lower than the dose that may be used for the direct tumor effect of compound A in existing monotherapy approaches.

[0107] In some embodiments, compound A is administered in amounts ranging from 0.1 mg or about 0.1 mg to 1 mg or about 1 mg. The dose can be administered daily over a cycling regimen. In some embodiments, the method provided is carried out by administering an amount of the compound that is 1 mg / day or less than 1 mg / day, for example, 0.9 mg, 0.8 mg, 0.7 mg, 0.6 mg, 0.5 mg, 0.4 mg, 0.3 mg, 0.2 mg, or 0.1 mg, or about 0.9 mg, 0.8 mg, 0.7 mg, 0.6 mg, 0.5 mg, 0.4 mg, 0.3 mg, 0.2 mg, or 0.1 mg, or any value between any of the above. In some embodiments, compound A is administered at 0.3 mg / day or about 0.3 mg / day. In some embodiments, compound A is administered at 0.45 mg / day or about 0.45 mg / day. In some embodiments, compound A is administered at a dose of 0.6 mg / day or approximately 0.6 mg / day.

[0108] In some embodiments, compound A is administered to the subject for a sufficient period of time after lymphocyte apheresis, thereby minimizing the myelosuppressive effects of compound A and lymphocyte apheresis.

[0109] In some embodiments, the method provided is used at a point in time when T cell therapy (e.g., CAR T cells) may or are likely to exhibit depletion characteristics. In some embodiments, the depletion phenotype is evident after T cells that have reached peak expansion begin to decrease in number in the subject's blood. In some embodiments, the method of exposing or contacting T cells (CAR T cells) for T cell therapy with compound A is performed at a point in time when the T cells show increased hypofunction or depletion compared to a point in time immediately before the T cells are exposed to the antigen (baseline) or a point in time when the cells have been exposed to the antigen but continue to proliferate and have not yet reached peak expansion. In some embodiments, the increase in hypofunction or depletion can be determined by an increase in the expression of depletion markers compared to an earlier point in time. In some embodiments, the increase in hypofunction or depletion, such as an increase in the expression of depletion markers, occurs at a point in time after administration of T cell therapy (e.g., CAR T cells) to a subject having a disease or condition related to the antigen targeted by the T cell therapy. T cells, such as T cells in the peripheral blood after administration to the target population, can be monitored for markers of T cell activation or depletion, including PD-1, TIM-3, and LAG-3.

[0110] In some embodiments, the method provided requires the administration of T cell therapy, e.g., CAR T cells, and the initiation of compound A administration at a point prior to the point in time when the CAR T cells exhibit or are likely to exhibit a depletion phenotype. In some embodiments, the administration of compound A is initiated at a point in time when the CAR T cells are still expanding or are still capable of expanding. In some embodiments, the administration of compound A is initiated at a point prior to the presence of a peak CAR T cell number in the subject's blood, or at a point suspected to be earlier. In some embodiments, the initiation of compound A administration at this point enhances CAR T cell function. In some embodiments, the initiation of compound A administration at this point also delays or prevents CAR T cell depletion.

[0111] In some embodiments, the administration of compound A is initiated when peak CAR-T cells are present in the subject's blood, or when they are suspected or likely to be present, or when they are highly suspected or likely to be present, for example, within 21 days after the start of T cell administration. In some cases, peak CAR-T cells are present within 11 to 15 days after the administration of CAR T cells. In some embodiments, the administration of compound A is initiated 1 to 15 days after the start of cell therapy, for example, 1 day or about 1 day, 8 days or about 8 days, or 15 days or about 15 days. In some embodiments, compound A is administered when the subject does not exhibit severe toxicity after the administration of cell therapy.

[0112] In some aspects, in any of the methods provided, administration of the compound is initiated (or initiated) within 21 days after administration of T-cell therapy and is carried out in a cycling regimen comprising: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to 3 consecutive weeks; a rest period of at least 1 week in which the compound is not administered, beginning at the end of the first administration period; and a second administration period comprising a 4-week cycle in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for 3 consecutive weeks over a 4-week period. In some aspects, the compound is administered at approximately 0.30 mg, 0.45 mg, or 0.60 mg / day during the first and second administration periods. In some aspects, during one or more of the 4-week cycles, the compound is not administered for 1 week after 3 consecutive weeks.

[0113] In some embodiments, the methods provided do not result in a high rate or likelihood of toxicity or toxic outcomes, such as neurotoxicity (NT), cytokine release syndrome (CRS), or hematological toxicity such as neutropenia, or reduce the rate or likelihood of toxicity or toxic outcomes compared to other specific cell therapies or immunomodulatory drug regimens.

[0114] 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, less than 50% of subjects exhibit neutropenia of grade 3 or higher, e.g., long-term grade 3 neutropenia or grade 4 neutropenia, and / or thrombocytopenia of grade 3 or higher, e.g., grade 3 or grade 4 thrombocytopenia. In some embodiments, at least 50% of subjects treated according to the method (e.g., at least 60%, at least 70%, at least 80%, at least 90%, or more of treated subjects) exhibit severe neutropenia or severe thrombocytopenia of grade 3 or higher.

[0115] In some embodiments, the method does not result in, or increases the risk of, severe NT (sNT), severe CRS (sCRS), macrophage activation syndrome, oncolytic syndrome, fever of at least 38°C or at least about 38°C for three days or more, and plasma levels of CRP of at least 20 mg / dL or at least about 20 mg / dL. In some embodiments, 30%, 35%, 40%, 50%, 55%, 60%, or more of subjects treated according to the provided method, or about 30%, 35%, 40%, 50%, 55%, 60%, or more, exhibiting no grade of CRS or any grade of neurotoxicity. In some embodiments, less than 50% 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) of grade 2 or higher and / or neurotoxicity of grade 2 or higher. In some embodiments, at least 50% of 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) will not exhibit serious toxic outcomes (e.g., severe CRS or severe neurotoxicity), for example, not exhibiting grade 3 or higher neurotoxicity, and / or not exhibiting severe CRS, or exhibiting these within a specific period after treatment, for example, within one week, two weeks, or one month after cell administration.

[0116] In some cases, compound A is administered at a time when it can efficiently / effectively boost or prime the cells. In some embodiments, administration of compound A is initiated when the peak or maximum level of cells in the cell therapy is detectable in the subject's blood or earlier. In some embodiments, the method provided can enhance T-cell therapy, e.g., CAR-T-cell therapy, which in some aspects can improve treatment outcomes. In some embodiments, the method is particularly advantageous in subjects where the cells in T-cell therapy show weak expansion, depletion, and decreased or diminished persistence, as well as in subjects with cancer that is resistant or refractory to other therapies and / or invasive or high-risk cancer.

[0117] In some embodiments, subjects receiving T-cell therapy, such as CAR-T cells, are monitored for the presence, absence, or level of therapeutic T cells in the subject in biological samples of the subject, such as in the subject's blood. In some embodiments, the method provided results in genetically modified cells having increased persistence and / or better potency in the subject to which it is administered. In some embodiments, the persistence of genetically modified cells, such as CAR-expressing T cells, in the subject is greater than the persistence that would be achieved by alternative methods, including the administration of T-cell therapy but in the absence of the administration of compound A. In some embodiments, the persistence increases by at least 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times, or more, or by at least about 1.5 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times, or more.

[0118] In some embodiments, the degree or extent of persistence of administered cells can be detected or quantified after administration to a subject. For example, in some cases, quantitative PCR (qPCR) is used to assess the amount of cells expressing recombinant receptors (e.g., CAR-expressing cells) in the blood or serum or organ or tissue (e.g., disease site) of a subject. In some embodiments, persistence is quantified as the number of copies of receptor-expressing cells, e.g., CAR-encoding DNA or plasmid per microgram of DNA, or per microliter of sample, e.g., blood or serum, or per microliter of peripheral blood mononuclear cells (PBMCs) or leukocytes or T cells. In some embodiments, flow cytometry assays can also be performed to detect receptor-expressing cells using antibodies that are generally specific to the receptor. Cell-based assays can also be used to detect the number or proportion of cells that can bind to and / or neutralize and / or induce a response to functional cells, e.g., disease or condition cells, or cells expressing antigens recognized by the receptor, and / or induce a response to them, e.g., a cytotoxic response. In any such configuration, the degree or level of expression of another marker associated with recombinant receptors (e.g., CAR-expressing cells) can be used to distinguish the administered cells from the target endogenous cells.

[0119] In some embodiments, compound A is administered for a period of time to enhance, increase, or optimize the persistence of the response. In some aspects, the methods provided are based on the observation that subjects who achieve or are in a state of complete remission (CR) in 3 months, for example, generally 6 months, are more likely to sustain the response for a longer period, for example, beyond 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after the completion of treatment or after achieving complete response (CR) for the first time after administration of combination therapy, or to survive or survive without progression for about 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some aspects, the methods are implemented to administer compound A for a period of at least 3 months, for example, at least 4 months, at least 5 months, or at least 6 months, after the initiation of T-cell therapy, such as in a specific cycling regimen as described. In some embodiments, compound A is administered for at least 6 months or at least 180 days after the initiation of T-cell therapy, such as in certain cycling regimens described. In some embodiments, at the end of the period, if the subject shows a complete response (CR), or if the disease or condition progresses or relapses in the subject after remission following treatment (combination therapy), administration of compound A is terminated or discontinued. In some aspects, continued administration of compound A may be performed in subjects showing a partial response (PR) or stable disease (SD) at the end of the period (e.g., 3 months or approximately 3 months, or 6 months or approximately 6 months). In other aspects, the period is fixed, and no further administration of compound A is performed.

[0120] In some aspects, the methods and uses provided offer or achieve an improved or more sustained response or effect compared to methods involving the administration of T-cell therapy or compound A, without administration as monotherapy or as combination therapy as described herein, in specific alternative methods, such as in specific groups of subjects to be treated. In some aspects, the method is advantageous by administering T-cell therapy, such as compositions containing cells for adoptive cell therapy, such as T-cell therapy (e.g., CAR-expressing T cells), and compound A. In some aspects, such a response is observed in high-risk patients with a poor prognosis, such as patients with high-risk diseases, such as patients with high-risk NHL. In some aspects, the method treats forms of B-cell non-Hodgkin lymphoma (NHL) that are highly malignant and / or have a poor prognosis, such as subjects with relapsed or refractory (R / R) NHL or with a poor prognosis. In some aspects, subjects treated according to the methods provided have diffuse large B-cell lymphoma (DLBCL) or follicular lymphoma.

[0121] 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 method and / or with the provided product, kit or composition achieve a complete response (CR). In some embodiments, subjects are in a CR state and exhibit minimal residual disease (MRD). In some embodiments, subjects are in a CR state and 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 method and / or with the provided product, kit or composition 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 method and / or using the provided product, kit, or composition achieve complete response (CR) or partial response (PR) at 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 1 year after the start of administration of cell therapy.

[0122] 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 method and / or with the provided product, kit or composition remain in a state of response, e.g., CR or objective response (OR), up to 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months or more after the initiation of administration of cell therapy. In some embodiments, such responses, such as CR or OR, persist for at least 60%, at least about 60%, at least 70%, at least 80%, at least 90%, at least 95%, or more of subjects treated according to the provided method, or for at least 3, 4, 5, or 6 months, or for at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more, in such subjects who achieve 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 method provided and / or using the product, kit or composition provided, or such subjects who achieve CR by 3 months, 4 months, 5 months or 6 months, survive or survive without progression for 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or longer, or approximately 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or longer.

[0123] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated by reference in whole for the same degree as each individual publication is incorporated by reference individually. If any definition provided herein is inconsistent with or contradicts any definition provided herein in a patent, patent application, published patent application, or other publication incorporated herein by reference, the definition provided herein shall prevail over the definition incorporated herein by reference.

[0124] Section headings used in this specification are for organizational purposes only and should not be construed as limiting the subjects described herein.

[0125] I. Combination Therapy For the treatment of subjects with cancer, engineered cells such as T cells (e.g., CAR-T cells) and (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione or formula I Methods and uses of pharmaceutically acceptable salts, solvates, hydrates, stereoisomers, tautomers, or racemic mixtures (compound A) of the compound TIFF0007837864000010.tif28128, or compositions thereof, are provided. In some embodiments, the methods are for treating subjects having B-cell malignancies. In some embodiments, the methods are for treating lymphomas such as leukemia or non-Hodgkin lymphoma (NHL). In some embodiments, the methods and uses provide or achieve an improved and / or more persistent response or effect, for example, in a particular group of subjects being treated, compared to a particular alternative method.

[0126] In some embodiments, the method and use include (1) administering a T-cell therapy to a subject that expresses a genetically modified cell surface receptor (e.g., recombinant antigen receptor), which is generally a chimeric receptor such as a chimeric antigen receptor (CAR), and recognizes an antigen associated with and / or specific to a B-cell malignancy such as leukemia or lymphoma (e.g., NHL), and / or a cell type from which it is derived; and (2) administering compound A to a subject. In some embodiments, the administration of compound A is initiated after (following the administration of) the T-cell therapy or after (following the initiation of) the administration of the T-cell therapy. In some cases, compound A is administered to a subject that has received T-cell therapy. The method generally includes administering one or more doses of cells and one or more doses of compound A to a subject.

[0127] For example, combination therapies comprising engineered cells expressing recombinant receptors such as chimeric antigen receptors (CARs) and compound A, or compositions comprising engineered cells and / or compound A as described herein, are useful in a variety of therapeutic, diagnostic, and prophylactic indications. For example, the combination is useful for treating a variety of diseases and disorders in a subject. Such methods and uses include, for example, therapeutic methods and therapeutic uses that involve administering a composition comprising engineered cells, compound A and / or one or both to a subject having a disease, condition, or disorder such as a tumor or cancer. In some embodiments, the composition comprising engineered cells, compound A and / or one or both is administered in an amount effective to result in the treatment of the disease or disorder. Uses include the use of a composition comprising engineered cells, compound A and / or one or both in such methods and therapies, and in the preparation of agents for carrying out such therapeutic methods. In some embodiments, the method is carried out by administering a composition comprising engineered cells, compound A and / or one or both to a subject having or suspected of having a disease or condition. In some embodiments, the method thereby treats a disease, condition, or disorder in a subject. In some embodiments, the manipulated cells are any of those described in Section II.

[0128] In some embodiments, combination therapy is administered to subjects having a specific B-cell malignancy. The B-cell malignancy to be treated may be any in which the expression of an antigen is related to and / or involved in the pathogenesis of the B-cell malignancy, e.g., causing, exacerbating, or otherwise involved in the B-cell malignancy. Exemplary B-cell malignancies may include diseases or conditions (e.g., cancer) associated with malignancy or cell transformation. Exemplary antigens, including antigens associated with a variety of B-cell malignancies that can be treated, are described herein. In certain embodiments, the chimeric antigen receptor specifically binds to the antigen associated with the disease or condition. In some embodiments, the antigen targeted by the receptor includes an antigen associated with a B-cell malignancy, such as one of several known B-cell markers. In some embodiments, the antigen is expressed by or on B cells, including human B cells. In some embodiments, the antigen targeted by the receptor is CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Ig kappa, Ig lambda, CD79a, CD79b, or CD30. In some embodiments, the antigen is CD19, and the chimeric antigen receptor binds specifically to CD19. In some embodiments, the CD19 antigen is human CD19. Any description of the method provided herein in which CAR-expressing T cells are specific to CD19 can also be carried out by targeting another B cell antigen, or an antigen associated with or expressed on cells of a T cell malignancy, for example, any of the above.

[0129] In some aspects, B-cell malignancies to be treated include leukemias and lymphomas, such as acute myeloid leukemia (AML), chronic myeloid (or myelogenous) leukemia (CML), acute lymphoblastic (or lymphoblastic) leukemia (ALL), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), anaplastic large cell lymphoma (ALCL), follicular lymphoma, refractory follicular lymphoma, and diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disease or condition is a B-cell malignancy selected from acute lymphoblastic leukemia (ALL), adult ALL, chronic lymphoblastic leukemia (CLL), non-Hodgkin lymphoma (NHL), and diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disease or condition is NHL, which is selected from the group consisting of invasive NHL, diffuse large B-cell lymphoma (DLBCL), NOS (de novo and painless transformed), primary mediastinal large B-cell lymphoma (PMBCL), T-cell / histiocyte-rich large B-cell lymphoma (TCHRBCL), Burkitt lymphoma, mantle cell lymphoma (MCL), and / or follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B).

[0130] In some embodiments, the method includes the step of treating a subject having a lymphoma or leukemia, such as non-Hodgkin lymphoma (NHL), by administering antigen receptor-expressing cells (e.g., CAR-expressing cells) and compound A. In some embodiments, compound A is administered after or following the administration of recombinant receptor-expressing cells (e.g., CAR-expressing cells), for example, after or following the commencement of administration of recombinant receptor-expressing cells (e.g., CAR-expressing cells).

[0131] In some aspects, NHL can be staging based on the Lugano classification (see, e.g., Cheson et al., (2014) JCO 32(27):3059-3067; Cheson, BD(2015) Chin Clin Oncol 4(1):5). In some cases, the stage is represented by Roman numerals I through IV (1-4), with E indicating a localized stage (I or II) lymphoma affecting extra-lymphatic organs (extra-lymphatic organs). Stage I represents involvement of one lymph node or a group of adjacent lymph nodes, or a single extra-lymphatic lesion (IE) without lymphatic involvement. Stage II represents involvement of two or more groups of lymph nodes on the same side of the septum, or involvement of a stage I or II lymphatic extent (IIE) with localized adjacent extra-lymphatic involvement. Stage III represents the involvement of lymph nodes on both sides of the diaphragm or above the diaphragm, with splenic involvement. Stage IV represents further extra-adjacent lymph node involvement. Additionally, "giant lesion" may be used to describe a large tumor in the chest, particularly in Stage II. The extent of the disease is determined by positron emission tomography (PET)-computed tomography (CT) in the case of Avid lymphoma, and by CT in the case of non-Avid histology.

[0132] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status index can be used to assess or select candidates for treatment, such as those with poor outcomes from previous treatments (see, e.g., Oken et al. (1982) Am J Clin Oncol. 5:649-655). In some embodiments, candidates have an ECOG status of less than 1 or 1. The ECOG performance status scale represents a patient's level of function in terms of their ability to care for themselves, daily living activities, and physical abilities (e.g., walking, working). In some embodiments, an ECOG performance status of 0 indicates that the candidate can perform normal activities. In some embodiments, a candidate with an ECOG performance status of 1 shows some limitations in physical activity, but the candidate is fully able to walk. In some embodiments, a candidate with an ECOG performance status of 2 is able to walk more than 50% of the time. In some cases, individuals with an ECOG performance status of 2 may also be able to care for themselves; see, for example, Sorensen et al., (1993) Br J Cancer 67(4)773-775. The criteria reflecting ECOG performance status are shown in Table 1 below.

[0133] (Table 1) ECOG Performance Status Criteria TIFF0007837864000011.tif56162

[0134] In some aspects, the subjects have had or been identified as having double / triple-hit lymphoma or lymphoma of a double / triple-hit molecular subtype. In some aspects, the lymphoma is a double-hit lymphoma characterized by the presence of MYC (myelocytoma oncogene), BCL2 (B-cell lymphoma 2), and / or BCL6 (B-cell lymphoma 6) gene rearrangements (e.g., translocations). In some aspects, the lymphoma is a triple-hit lymphoma characterized by the presence of MYC, BCL2, and BCL6 gene rearrangements; see, e.g., Aukema et al., (2011) Blood 117:2319-2331. In some aspects of such aspects, the subjects are ECOG 0-1. In multiple aspects, the treatment is indicated for such subjects, and / or the instructions indicate administration to subjects within such populations. In some aspects, based on the 2016 WHO criteria (Swerdlow et al., (2016) Blood 127(20):2375-2390), double / triple-hit lymphoma can be considered a high-grade B-cell lymphoma with MYC and BCL2 and / or BCL6 rearrangements accompanied by DLBCL histology (double / triple-hit).

[0135] In some embodiments, combination therapy is administered to subjects who are poorly responding to, likely to be poorly responding to, or predicted to be poorly responding to, and / or do not respond, are likely to be unresponsive, and / or are predicted to be unresponsive, or do not respond within a certain time and / or to a certain degree. In some embodiments, combination therapy is administered to subjects who do not show, are likely to not show, or are predicted to not show, a complete or overall response within one month, two months, or three months after the start of cell therapy administration, etc. In some embodiments, combination therapy is administered to subjects who show, are likely to show, or are predicted to show progression (PD) within one month, two months, or three months after the administration of cell therapy, etc. In some embodiments, subjects are likely to show or are predicted to show no response or a particular response based on multiple similar subjects who have been treated or previously treated with cell therapy.

[0136] In some embodiments, the method provided includes a step of treating a specific group or subset of subjects, e.g., subjects identified as having a high-risk disease, e.g., high-risk NHL. In some embodiments, the method treats subjects with highly aggressive and / or poorly prognosed forms of B-cell non-Hodgkin lymphoma (NHL), e.g., subjects with relapsed or refractory (R / R) or poorly prognosed NHL. In some cases, the overall response rate (ORR) to available therapies, standard therapies, or reference therapies for the disease and / or patient population to which the treatment is indicated is less than 40%, and / or the complete response (CR) is less than 20%. In some embodiments, in chemotherapy-resistant DLBCL, the ORR to reference or available therapies or standard therapies is approximately 26%, and the CR is approximately 8% (Crump et al. Outomes in refractory aggressive diffuse large B-cell lymphoma (DLBCL): Results from the international SCHOLAR study. ASCO 2016 [Abstract 7516]). In some aspects, the methods, compositions, uses, and products provided achieve superior responses that are better than available treatments.

[0137] In some embodiments, methods and uses for the treatment of subjects described herein include the step of selecting or identifying a specific group or subset of subjects based, for example, on a particular type of disease, diagnostic criteria, prior treatment and / or response to prior treatment. In some embodiments, the method includes the step of treating subjects who have relapsed or become refractory after remission following treatment with one or more prior therapies, or subjects who have relapsed or become refractory (R / R) to one or more lines of standard treatment, including one or more prior therapies, for example, those described herein.

[0138] In some embodiments, the subject has received more than 1, 2, 3, 4, 5, or 6 previous treatments. In some embodiments, the subject has received 1 previous treatment. In some embodiments, the subject has received approximately 2 to 4 previous treatments. In some embodiments, the subject has received approximately 5 to 6 previous treatments. In some embodiments, the subject has received more than 6 previous treatments.

[0139] In some embodiments, the subject has previously been treated with a therapy or treatment targeting B-cell malignancies, e.g., NHL, prior to the administration of cells expressing recombinant receptors. In some embodiments, the subject has previously been treated with cell therapy (e.g., CAR+T cells). In some embodiments, the subject has previously been treated with hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some embodiments, the subject has had a poor prognosis after treatment with standard therapy and / or has failed one or more previous lines of treatment. In some embodiments, the subject has been treated, or has previously received, at least one or about one or about one, at least two or about two or about two, at least three or about three or about three, at least four or at least four or about four, at least five or at least five or about five, at least six or at least six or about six, or at least seven or at least seven or about seven other therapies to treat NHL, other than lymphocyte apheresis. In some embodiments, the subject has previously been treated with chemotherapy or radiotherapy. In some aspects, the subject is refractory or unresponsive to other therapies or medications. In some aspects, the subject has persistent or relapsing disease after treatment with another therapy or therapeutic intervention, including, for example, chemotherapy or radiation.

[0140] In some embodiments, the combination therapy is administered to subjects whose condition has progressed with prior treatment. In some embodiments, the combination therapy is administered to subjects who have ceased responding to prior therapy. In some embodiments, the combination therapy is administered to subjects who have relapsed after remission following prior treatment. In some embodiments, the combination therapy is administered to subjects who are refractory to prior treatment. In some embodiments, the combination therapy is administered to subjects who have not achieved an optimal response (e.g., complete response, partial response, or stable disease) to prior therapy.

[0141] In some embodiments, the subject is refractory to the previous last treatment. In some embodiments, the subject has relapsed to the previous last treatment. The condition is refractory if the subject achieved a response of less than a partial response to the previous last treatment. In some embodiments, the subject has previously received chemotherapy. In some embodiments, the subject is chemotherapy-resistant to the previous chemotherapy. In some embodiments, the subject is chemosensitive to the previous therapy. The condition is chemotherapy-resistant if the subject achieved stable disease (SD) or progressive disease (PD) to the regimen including the last chemotherapy, or relapsed less than 12 months after autologous stem cell transplantation. Otherwise, the condition is chemosensitive.

[0142] In some embodiments, the prior treatment or therapy includes a CD20-targeting agent. In some embodiments, the prior treatment or therapy includes an anthracycline. In some embodiments, the prior treatment or therapy includes cell therapy (e.g., T-cell therapy, e.g., CAR T-cell therapy).

[0143] In some embodiments, the Method, Use and Product include or are used for the treatment of a subject, including the step of selecting or identifying a specific group or subset of subjects, such as any group of subjects described, based on, for example, a particular type of disease, diagnostic criteria, prior treatment and / or response to prior treatment. In some embodiments, the Method includes the step of treating a subject who has relapsed after remission or has become refractory to therapy after treatment with one or more prior therapies, or a subject who has relapsed or is refractory (R / R) to one or more prior therapies, such as one or more lines of standard treatment, such as cell therapy (e.g., CAR+T cell). In some embodiments, the Method includes the step of treating a subject having diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS; de novo and transformed from painless), primary mediastinal (thymic) large B-cell lymphoma (PMBCL) or follicular lymphoma grade 3B (FL3B), EBV-positive DLBCL, or EBV-positive NOS. In some embodiments, the method includes a step of treating subjects having an Eastern Collaborative Oncology Group Performance Status (ECOG) of less than 1, e.g., 0 to 1. In some embodiments, the method treats a poor-prognosed population of DLBCL patients or subjects that generally have poor response to therapy or a particular reference therapy, e.g., those with one or more, e.g., two or three chromosomal translocations (e.g., so-called “double-hit” or “triple-hit” lymphomas, such as high-grade B-cell lymphomas with DLBCL histology and MYC and BCL2 and / or BCL6 rearrangements; usually those with the translocated MYC / 8q24 locus in combination with the t(14;18)(q32;q21)bcl-2 gene or / and the BCL6 / 3q27 chromosomal translocation; e.g., see Xu et al. (2013) Int J Clin Exp Pathol. 6(4):788-794), and / or relapsed, optionally relapsed within 12 months, and / or considered chemotherapy-resistant.

[0144] In some embodiments, subjects have germinal center-like (GCB) DLBCL. In some embodiments, subjects have non-germinal center-like (non-GCB) DLBCL. In some embodiments, subjects have double-hit lymphoma (DHL). In some embodiments, subjects have triple-hit lymphoma (THL). In some embodiments, subjects are positive for gene expression indicating responsiveness to treatment with compound A. In some embodiments, subjects are negative for the expression of the said gene. See Blood 2017 130:4118.

[0145] In some embodiments, the antigen receptor (e.g., CAR) specifically binds to a target antigen associated with a disease or condition, such as NHL. In some embodiments, the antigen associated with the disease or disorder is selected from CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igκ, Igλ, CD79a, CD79b, or CD30. In some embodiments, the antigen is CD19. In some embodiments, the CD19 antigen is human CD19.

[0146] In some embodiments, the method comprises cell therapy and administration of compound A to subjects who have, are at risk of having, or are suspected of having a B-cell malignancy.

[0147] In some embodiments, the method involves administering cells to subjects selected or identified as having a specific prognosis or risk of NHL. Non-Hodgkin lymphoma (NHL) can be a diverse disease. Some subjects with NHL may survive without treatment, while others may require immediate intervention. In some cases, subjects with NHL may be classified into groups that may inform the prognosis of the disease and / or recommended treatment strategies. In some cases, these groups may be “low-risk,” “intermediate-risk,” “high-risk,” and / or “very high-risk,” and patients may be classified so depending on many factors, including but not limited to genetic abnormalities and / or morphological or physical characteristics. In some embodiments, subjects treated according to the method and / or according to the product or composition are classified or identified based on their risk of NHL. In some embodiments, subjects are those with high-risk NHL.

[0148] In some embodiments, the subjects to be treated include those with invasive NHL, particularly diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS; de novo and transformed from painless), T-cell / histiocyte-rich large B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma (PMBCL), follicular lymphoma grade 3B (FL3B), EBV-positive DLBCL, EBV-positive NOS, or high-grade B-cell lymphoma ("double-hit" or "triple-hit" lymphoma) with MYC and BCL2 and / or BCL6 rearrangements with DLBCL histology. In some embodiments, the subject's disease was relapsed or refractory to at least two prior lines of treatment. In some embodiments, prior treatment included CD20-targeted agents and / or anthracyclines. In some embodiments, the subject had an ECOG score of 0–1 at screening. In some embodiments, subjects have a disease that is positive for positron emission tomography (PET) according to the Lugano classification (Cheson, 2014). In some embodiments, subjects may optionally have been previously treated with allogeneic stem cell transplantation (SCT).

[0149] In some embodiments, the subjects are adults. In some embodiments, the subjects are male. In some embodiments, the subjects are female. In some embodiments, the subjects are at least 40 years old at the time of administration of the combination therapy (e.g., at the time of administration of cell therapy). In some embodiments, the subjects are under 40 years old at the time of administration of the combination therapy (e.g., at the time of administration of cell therapy). In some embodiments, the subjects are approximately 40 to 65 years old at the time of administration of the combination therapy (e.g., at the time of administration of cell therapy). In some embodiments, the subjects are at least 65 years old at the time of administration of the combination therapy (e.g., at the time of administration of cell therapy).

[0150] A. Administration of cell therapy Methods for administering cells for adoptive cell therapy are publicly known and may be used in connection with the methods, compositions, and products provided. For example, methods for adoptive T cell therapy are described in, for example, U.S. Patent Application Publication No. 2003 / 0170238 by Gruenberg et al.; U.S. Patent No. 4,690,915 by Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85). See, for example, Themeli et al. (2013) Nat Biotechnol. 31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; and Davila et al. (2013) PLoS ONE 8(4):e61338.

[0151] In some embodiments, cells used in or administered in connection with the provided method contain or are engineered to contain engineered receptors, such as chimeric antigen receptors (CARs) or T cell receptors (TCRs). Among the compositions are pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy. Therapeutic methods are also provided for administering cells and compositions to a subject, such as a patient, according to the provided method and / or according to the provided product or composition.

[0152] Cells generally express recombinant receptors, antigen receptors including functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs), and other antigen-binding receptors such as transgenic T cell receptors (TCRs). Some receptors are other chimeric receptors. Exemplary engineered cells for administration as cell therapy in the manner provided are described in Section II.

[0153] In some embodiments, cell therapy, such as adoptive T-cell therapy, is carried out by autologous transplantation, where cells are isolated and / or prepared from a subject to receive cell therapy, or from a sample derived from such a subject. Thus, in some aspects, the cells originate from a subject in need of treatment, such as a patient, and are administered to the same subject after isolation and processing.

[0154] In some embodiments, cell therapy, such as adoptive T-cell therapy, is carried out by allogeneic transplantation, in which cells are isolated and / or 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.

[0155] T-cell therapy cells can be administered in a composition formulated for administration, or in two or more compositions (e.g., two compositions) formulated for separate administrations. The cell dose(s) may include a specific or relative number of cells or engineered cells, and / or a defined ratio or composition of two or more subtypes in the composition, such as CD4 vs. CD8 T cells.

[0156] Cells can be administered by any suitable means, such as bolus injection, by injection, such as intravenous or subcutaneous injection, intraocular injection, periorbital injection, subretinal injection, intravitreous injection, transseptal injection, subscleral injection, choroidal injection, anterior chamber injection, subconjunctival injection, subconjunctival injection, sub-Tenon's capsule injection, retrobulbar injection, peribulbar injection, or posterior parascleral delivery. In some embodiments, they are administered by parenteral administration, intrapulmonary administration, and intranasal administration, and, if desired for local treatment, by intrafocal administration. Parenteral administration includes intramuscular, intravenous, intra-arterial, 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 bolus administrations of cells over a period of, for example, three days or less, or by continuous infusion of cells. In some embodiments, the administration of cellular doses or any further therapies, such as lymphocyte apheresis, interventional therapy, and / or combination therapy, is carried out by outpatient delivery.

[0157] For the treatment of a disease, the appropriate dosage may depend on the type of disease being treated, the type of cells or recombinant receptor, the severity and course of the disease, previous therapies, the subject's medical history and response to the cells, and the discretion of the attending physician. The composition and cells are administered appropriately to the subject, in some embodiments, either in a single dose or over a series of treatments.

[0158] Pre-conditioning subjects with immunodepletion therapy (e.g., lymphocyte apheresis) can, in some aspects, improve the effectiveness of adoptive cell therapy (ACT).

[0159] Therefore, in some embodiments, the method includes administering a pre-treatment agent to a subject prior to the initiation of cell therapy, such as a lymphocyte depletion agent or chemotherapeutic agent, e.g., cyclophosphamide, fludarabine, or a combination thereof. For example, the subject may be administered the pre-treatment agent at least two days before the initiation of cell therapy, e.g., at least three days, at least four days, at least five days, at least six days, or at least seven days prior. In some embodiments, the subject is administered the pre-treatment agent within seven days before the initiation of cell therapy, e.g., within six days, at least five days, at least four days, at least three days, or within two days.

[0160] In some embodiments, subjects are pre-treated with cyclophosphamide at doses of 20 mg / kg to 100 mg / kg or approximately 20 mg / kg to 100 mg / kg, for example, 40 mg / kg to 80 mg / kg or approximately 40 mg / kg to 80 mg / kg. In some embodiments, subjects are pre-treated with 60 mg / kg or approximately 60 mg / kg of cyclophosphamide. In some embodiments, cyclophosphamide may be administered as a single dose or in multiple doses, such as daily, every other day, or every three days. In some embodiments, cyclophosphamide is administered once daily for one or two days. In some embodiments, if the lymphocyte depletion agent contains cyclophosphamide, subjects receive 100 mg / m² 2 ~500mg / m 2 Alternatively, approximately 100 mg / m² 2 ~500mg / m 2 For example, 200 mg / m² 2 ~400mg / m 2 Alternatively, approximately 200 mg / m² 2 ~400mg / m 2 , or 250 mg / m² 2 ~350mg / m 2 Alternatively, approximately 250 mg / m² 2 ~350mg / m 2 Cyclophosphamide is administered at doses (including both extreme values). In some cases, the subject is approximately 300 mg / m². 2 The patient is administered cyclophosphamide. In some cases, the patient receives approximately 500 mg / m². 2Cyclophosphamide is administered. In some embodiments, cyclophosphamide may be administered as a single dose or as multiple doses, such as daily, every other day, or every three days. In some embodiments, cyclophosphamide is administered daily, for example, for 1 to 5 days, or for example, for 3 to 5 days. In some cases, the subject receives approximately 300 mg / m² before initiating cell therapy. 2 Cyclophosphamide is administered daily for three days. In some cases, the subject receives approximately 500 mg / m² before initiating cell therapy. 2 The patient is administered cyclophosphamide daily for three days.

[0161] In some embodiments, when the lymphocyte depletion agent contains fludarabine, the target is 1 mg / m². 2 ~100mg / m 2 Alternatively, approximately 1 mg / m² 2 ~100mg / m 2 For example, 10 mg / m² 2 ~75mg / m 2 Alternatively, approximately 10 mg / m² 2 ~75mg / m 2 , 15 mg / m² 2 ~50mg / m 2 Alternatively, approximately 15 mg / m² 2 ~50mg / m 2 , 20 mg / m² 2 ~40mg / m 2 Alternatively, approximately 20 mg / m² 2 ~40mg / m 2 , or 24 mg / m² 2 ~35mg / m 2 Alternatively, approximately 24 mg / m² 2 ~35mg / m 2 Fludarabine is administered at doses (including both extreme values). In some cases, the subject is approximately 30 mg / m². 2Fludarabine is administered. In some embodiments, fludarabine may be administered as a single dose or as multiple doses, such as daily, every other day, or every three days. In some embodiments, fludarabine is administered daily, for example, for 1 to 5 days, or for example, for 3 to 5 days. In some cases, the subject receives approximately 30 mg / m² before initiating cell therapy. 2 Fludarabine is administered daily for three days.

[0162] In some embodiments, the lymphocyte depletion agent includes a combination of agents such as a combination of cyclophosphamide and fludarabine. Therefore, the combination of agents may include cyclophosphamide in any dose or administration schedule as described above, and fludarabine in any dose or administration schedule as described above. For example, in some aspects, the subject is given 60 mg / kg (~2 g / m²) before the first dose or subsequent doses. 2 ) Cyclophosphamide and 25 mg / m² in 3-5 doses 2 Fludarabine is administered. In some embodiments, the subject is given 300 mg / m² before the start of cell therapy. 2 Cyclophosphamide and 30 mg / m² 2 Both fludarabine are administered daily for three days. In some embodiments, subjects receive 500 mg / m² before the initiation of cell therapy. 2 Cyclophosphamide and 30 mg / m² 2 Both fludarabine drugs are administered daily for three days.

[0163] Following cell administration, in some embodiments, the biological activity of the engineered cell population is measured by, for example, one of many known methods. Parameters to be evaluated include the specific binding of engineered or native 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 engineered cells to destroy target cells can be measured using any suitable known method, for example, by 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 cells is measured by testing 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 evaluating clinical outcomes such as a reduction in tumor burden or tumor volume.

[0164] 1. Compositions and Formulations In some embodiments, the dose of cells for cell therapies, such as T-cell therapy, which involves recombinant antigen receptors, for example, cells manipulated with CAR or TCR, is provided as a composition or formulation, such as a pharmaceutical composition or formulation. Such compositions can be used in the treatment of B-cell malignancies, etc., according to the methods and / or products or compositions provided.

[0165] The term "pharmaceutical preparation" refers to a preparation in which the biological activity of the active ingredient contained therein is effective, and which does not contain any further ingredients that would be unacceptably toxic to the subject to which the preparation is administered.

[0166] A "pharmaceutically acceptable carrier" refers to a component in a pharmaceutical preparation other than the active ingredient that is non-toxic to the target substance. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0167] In some embodiments, cell therapies, such as those involving 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 specific cells or agent and / or the method of administration. Thus, a variety of suitable formulations exist. For example, a pharmaceutical composition may contain a preservative. Suitable preservatives 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 a mixture thereof is 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). Pharmacochemically acceptable carriers are generally non-toxic to the recipient at the doses and concentrations used and include, but are not limited to, the following: buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, alkylparabens such as butyl or benzyl alcohol, methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol and m-cresol); low-molecular-weight Polypeptides (less than approximately 10 residues); proteins such as serum albumin, gelatin, or immunoglobulins; 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 nonionic surfactants such as polyethylene glycol (PEG).

[0168] In some cases, buffering agents are 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 cases, mixtures of two or more buffering agents are 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 administerable pharmaceutical compositions are known. Exemplary methods are described in more detail, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0169] The formulation may include an aqueous solution. The formulation or composition may also contain multiple active ingredients useful for a specific indication, disease, or condition treated with the cells or agent, provided that their respective activities do not adversely affect each other. Such active ingredients are appropriately present in combination in amounts effective for the intended purpose. Thus, in some embodiments, the pharmaceutical composition may further include other pharmaceutically active agents or drugs such as chemotherapeutic agents, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0170] In some embodiments, the pharmaceutical composition contains a quantity of cells effective for treating a disease or condition, such as a therapeutically effective or prophylactically effective amount. In some embodiments, therapeutic efficacy is monitored by periodic evaluation of the subject being treated. For repeated administrations over several days or longer, treatment is repeated, depending on the condition, until the desired suppression of disease symptoms occurs. However, other administration regimens may also be useful and may be determined. The desired dose can be delivered by a single bolus administration of the composition, by multiple bolus administrations of the composition, or by continuous infusion administration of the composition.

[0171] Cells may be administered using standard administration techniques, formulations, and / or apparatus. Formulations and apparatus, such as syringes and vials, are provided for the storage and administration of compositions. With respect to cells, administration may be autologous or heterologous. For example, immune-responsive cells or precursors may be obtained from one subject and administered to the same or different, compatible subjects. Peripheral blood-derived immune-responsive cells or their offspring (e.g., in vivo, ex vivo, or in vitro-derived) may be administered by catheter administration, systemic infusion, local infusion, intravenous injection, or local infusion including parenteral administration. When administering therapeutic compositions (e.g., pharmaceutical compositions containing genetically modified immune-responsive cells), therapeutic compositions are generally formulated in unit-dose injection forms (solutions, suspensions, emulsions).

[0172] Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, oral, sublingual, or suppository administration. In some embodiments, the agent or cell population is administered parenterally. As used herein, the term “parenteral” includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. In some embodiments, the agent or cell population is administered to a subject by peripheral systemic delivery via intravenous, intraperitoneal, or subcutaneous injection.

[0173] In some embodiments, compositions are provided as sterile liquid formulations, such as isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which can be buffered to a selected pH in some aspects. Liquid formulations are generally easier to prepare than gels, other viscous compositions, and solid compositions. Furthermore, liquid compositions are somewhat more convenient to administer, particularly by injection. On the other hand, viscous compositions can be formulated within a suitable viscosity range that provides a longer contact period with specific tissues. Liquid or viscous compositions may contain a carrier, which may be a solvent or dispersion medium containing, for example, water, saline solution, phosphate-buffered saline solution, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol) and suitable mixtures thereof.

[0174] Sterile injection solutions can be prepared by incorporating cells into a solvent such as a mixture with a suitable carrier, diluent, or excipient, such as sterile water, physiological saline, glucose, or dextrose.

[0175] Preparations used for in vivo administration are generally sterile. Sterility can be easily achieved, for example, by filtering through a sterile filtration membrane.

[0176] 2. Medication In some embodiments, the dose of cells is administered to the subject according to the method provided and / or according to the product or composition provided. In some embodiments, the size or timing of the dose is determined according to the specific disease or condition in the subject (e.g., cancer, e.g., B-cell malignancy). In some cases, the size or timing of the dose for a particular disease may be determined empirically, taking into consideration the explanation provided.

[0177] In some embodiments, the cell dose is 2 × 10 5 cells / kg or approximately 2 x 10⁻⁶ 5 From cells / kg to 2 × 10 6 cells / kg or approximately 2 x 10⁻⁶ 6 cells / kg, e.g., 4 × 10⁻⁶ 5 cells / kg or approximately 4 x 10⁻⁶ 5 From cells / kg to 1 × 10 6 cells / kg or approximately 1 x 10⁻⁶ 6 cells / kg, or 6 × 10⁶ 5 cells / kg or approximately 6 x 10⁻⁶ 5 8 x 10 cells / kg 5 cells / kg or approximately 8 x 10⁻⁶ 5 Contains cells / kg. In some embodiments, the dose of cells is 2 × 10⁶ (cells / kg) per kilogram of body weight of the subject. 5 The following cells (e.g., antigen-expressing cells such as CAR-expressing cells), e.g., 3 × 10 5 Cells / kg or less, or approximately 3 x 10⁻⁶ 5 cells / kg, 4×10 5 Cells / kg or less or approximately 4 x 10 5Cells / kg or less, 5×10 5 Cells / kg or less or approximately 5 x 10 5 Cells / kg or less, 6×10 5 Cells / kg or less, or approximately 6 x 10⁻⁶ 5 Cells / kg or less, 7×10 5 Cells / kg or less, or approximately 7 x 10 5 Cells / kg or less, 8×10 5 Cells / kg or less, or approximately 8 x 10 5 Cells / kg or less, 9×10 5 Cells / kg or less, or approximately 9 x 10 5 Cells / kg or less, 1×10 6 Cells / kg or less, or approximately 1 x 10⁻⁶ 6 Cells / kg or less, or 2 × 10⁻⁶ 6 Cells / kg or less, or approximately 2 x 10⁻⁶ 6 Contains cells / kg or less. In some embodiments, the cell dose is at least 2 × 10⁶ (cells / kg) per kilogram of body weight of the subject. 5 cells / kg (e.g., antigen-expressing cells such as CAR-expressing cells) or at least about 2 × 10⁻⁶ 5 cells / kg or 2 × 10⁻⁶ 5 cells / kg or approximately 2 x 10⁻⁶ 5 cells / kg, e.g., at least 3 × 10⁶ 5 cells / kg or at least approximately 3 × 10⁻⁶ 5 cells / kg or 3 × 10⁻⁶ 5 cells / kg or approximately 3 x 10⁻⁶ 5 cells / kg, at least 4 × 10⁻⁶ 5 cells / kg or at least approximately 4 × 10⁻⁶ 5 cells / kg or 4×10 5 cells / kg or approximately 4 x 10⁻⁶ 5 Cells / kg, at least 5 × 10⁻⁶ 5 cells / kg or at least approximately 5 × 10⁻⁶ 5 cells / kg or 5 x 10 5 cells / kg or approximately 5 x 10 5 Cells / kg, at least 6 × 10⁶ 5 cells / kg or at least approximately 6 × 10⁻⁶ 5 cells / kg or 6 x 10 5 cells / kg or approximately 6 x 10⁻⁶5 Cells / kg, at least 7 × 10⁶ 5 cells / kg or at least approximately 7 × 10⁻⁶ 5 cells / kg or 7×10 5 cells / kg or approximately 7 x 10⁻⁶ 5 Cells / kg, at least 8 × 10⁶ 5 cells / kg or at least approximately 8 × 10⁻⁶ 5 cells / kg or 8 x 10 5 cells / kg or approximately 8 x 10⁻⁶ 5 Cells / kg, at least 9 × 10⁶ 5 cells / kg or at least approximately 9 × 10⁶ 5 cells / kg or 9 x 10 5 cells / kg or approximately 9 x 10 5 cells / kg, at least 1 × 10⁶ 6 cells / kg or at least about 1 × 10⁻⁶ 6 cells / kg or 1 × 10⁻⁶ 6 cells / kg or approximately 1 x 10⁻⁶ 6 cells / kg, or at least 2 × 10⁶ 6 cells / kg or at least about 2 × 10⁻⁶ 6 cells / kg or 2 × 10⁻⁶ 6 cells / kg or approximately 2 x 10⁻⁶ 6 Contains cells / kg.

[0178] In certain embodiments, individual populations of cells, or subtypes of cells, range from approximately 1 million to approximately 100 billion cells and / or that amount of cells per kilogram of body weight of the subject, for example, 1 million to approximately 50 billion cells (e.g., approximately 5 million cells, approximately 25 million cells, approximately 500 million cells, approximately 1 billion cells, approximately 5 billion cells, approximately 20 billion cells, approximately 30 billion cells, approximately 40 billion cells, or a range defined by any two of the above values), for example, approximately 10 million to approximately 100 billion cells (e.g., approximately 20 million cells, approximately 30 million cells, approximately 40 million cells, approximately 60 million cells, approximately 70 million cells, approximately 80 million cells) The cells administered to the subject are approximately 90 million cells, approximately 10 billion cells, approximately 25 billion cells, approximately 50 billion cells, approximately 75 billion cells, approximately 90 billion cells (or a range defined by any two of the above values), and in some cases, approximately 100 million to approximately 50 billion cells (e.g., approximately 120 million cells, approximately 250 million cells, approximately 350 million cells, approximately 450 million cells, approximately 650 million cells, approximately 800 million cells, approximately 900 million cells, approximately 3 billion cells, approximately 30 billion cells, approximately 45 billion cells), or any value within these ranges and / or that amount of cells per kilogram of the subject's body weight. The dosage may vary depending on the disease or disorder and / or attributes specific to the patient and / or other treatment.

[0179] In some embodiments, the cell dose is a uniform or fixed dose of cells, such that the cell dose is not tied to or based on the subject's body surface area or weight.

[0180] In some embodiments, for example, when the subject is human, the dose is approximately 5 × 10 8 Cells expressing less than 1 × 10⁻¹ total recombinant receptors (e.g., CARs), T cells, or peripheral blood mononuclear cells (PBMCs), e.g., about 1 × 10⁻¹⁶ cells. 6 ~5×10 8 Such cells, for example, 2 × 10 6 , 5×10 6 , 1 x 10 7 , 5×10 7, 1 x 10 8 , 2×10 8 , 3 x 10 8 , or 4 x 10 8 The range of such total cells, or such cells in a range between any two of the aforementioned values. In some embodiments, if the subject is human, the dose is approximately 1 × 10⁻⁶ 6 ~3×10 8 Total recombinant receptor (e.g., CAR) expressing cells, e.g., about 1 × 10⁶ 7 ~2×10 8 Such cells, for example, 1 × 10 7 , 5×10 7 , 1 x 10 8 Or 1.5 × 10 8 The range of such total cells, or such cells in a range between any two of the aforementioned values. In some embodiments, the patient is administered multiple doses, and each dose or total dose may be within any of the aforementioned values. In some embodiments, the dose of cells is 1 × 10⁻⁶ 5 ~5×10 8 Or approximately 1 x 10 5 ~5×10 8 Total recombinant receptor (e.g., CAR) expressing T cells or total T cells, 1 × 10 5 ~1 × 10 8 Or approximately 1 x 10 5 ~1 × 10 8 Total recombinant receptor (e.g., CAR) expressing T cells or total T cells, 5 × 10 5 ~1 × 10 7 Or approximately 5 x 10 5 ~1 × 10 7 Total recombinant receptor (e.g., CAR) expressing T cells or total T cells, or 1 × 10⁶ 6 ~1 × 10 7 Or approximately 1 x 10 6 ~1 × 10 7 This includes administration of total recombinant receptor (e.g., CAR) expressing T cells or total T cells (including values ​​at both ends of the spectrum).

[0181] In some embodiments, the dose of T cells comprises CD4+ T cells, CD8+ T cells, or CD4+ and CD8+ T cells.

[0182] In some embodiments, for example, when the subject is human, the dose of CD8+ T cells, including CD4+ and CD8+ T cells, is approximately 1 × 10⁻⁶ 6 ~1 × 10 8 Total recombinant receptor (e.g., CAR) expressing CD8+ cells, e.g., approximately 5 × 10⁻⁶ 6 ~1 × 10 8 Such cells in the range of 1 × 10 7 , 2.5×10 7 , 5×10 7 , 7.5×10 7 Or 1 x 10 8 The range of such total cells, or such cells in a range between any two of the aforementioned values. In some embodiments, the patient is administered multiple doses, and each dose or total dose may be within any of the aforementioned values. In some embodiments, the dose of cells is 1 × 10⁻⁶ 7 ~0.75 × 10 8 Or approximately 1 x 10 7 ~0.75 × 10 8 Total recombinant receptor-expressing CD8+ T cells, 1 × 10 7 ~2.5×10 7 Or approximately 1 x 10 7 ~2.5×10 7 Total recombinant receptor-expressing CD8+ T cells, 1 × 10 7 ~0.75 × 10 8 Or approximately 1 x 10 7 ~0.75 × 10 8 This involves the administration of total recombinant receptor-expressing CD8+ T cells (including values ​​at both ends). In some embodiments, the cell dose is 1 × 10⁶ 7 Or approximately 1 x 10 7 , 2.5×10 7 Or approximately 2.5 x 10 7 , 5×10 7 Or approximately 5 x 10 7 , 7.5×10 7 Or approximately 7.5 x 10 7 , or 1 × 10 8 Or approximately 1 x 10 8 This includes administration of total recombinant receptor-expressing CD8+ T cells.

[0183] In some embodiments, for example, when the subject is human, the dose of CD4+ T cells, including a dose containing CD4+ and CD8+ T cells, is approximately 1 × 10⁻⁶ 6 ~1 × 10 8 Total recombinant receptor (e.g., CAR) expressing CD4+ cells, e.g., approximately 5 × 10⁻⁶ 6 ~1 × 10 8 Such cells, for example, 1 × 10 7 , 2.5×10 7 , 5×10 7 , 7.5×10 7 , or 1 x 10 8 The range of such total cells, or such cells in a range between any two of the aforementioned values. In some embodiments, the patient is administered multiple doses, and each dose or total dose may be within any of the aforementioned values. In some embodiments, the dose of cells is 1 × 10⁻⁶ 7 ~0.75 × 10 8 Or approximately 1 x 10 7 ~0.75 × 10 8 Total recombinant receptor-expressing CD4+ T cells, 1 × 10 7 ~2.5×10 7 Or approximately 1 x 10 7 ~2.5×10 7 Total recombinant receptor-expressing CD4+ T cells, 1 × 10 7 ~0.75 × 10 8 Or approximately 1 x 10 7 ~0.75 × 10 8 This involves the administration of total recombinant receptor-expressing CD4+ T cells (including values ​​at both ends). In some embodiments, the cell dose is 1 × 10⁶ 7 Or approximately 1 x 10 7 , 2.5×10 7 Or approximately 2.5 x 10 7 , 5×10 7 Or approximately 5 x 10 7 , 7.5×10 7 Or approximately 7.5 x 10 7 , or 1 × 10 8 Or approximately 1 x 10 8 This includes administration of total recombinant receptor-expressing CD4+ T cells.

[0184] In some embodiments, the dose of cells (e.g., recombinant receptor-expressing T cells) is administered to the subject as a single dose, or only once within a period of 2 weeks, 1 month, 3 months, 6 months, 1 year, or longer.

[0185] In the context of adoptive cell therapy, a given “dose” includes the administration of a given amount or number of cells as a single composition, and / or a single, uninterrupted administration, such as a single injection or continuous infusion, and also includes the administration of a given amount or number of cells as a divided dose or as multiple compositions, provided in multiple individual compositions or infusions over a specified period, such as three days or less. Thus, in some situations, the dose is a single or continuous administration of a specified number of cells given or initiated at a single point in time. However, in some situations, the dose is administered by 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 period of one day.

[0186] Therefore, in some aspects, a dose of cells is administered in a single pharmaceutical composition. In some aspects, a dose of cells is administered in multiple compositions that collectively contain a dose of cells.

[0187] The term "divided dose" refers to a dose that is divided to be administered over more than one day. This type of administration is included in the method of the present invention and is considered a single dose.

[0188] Therefore, the dose of cells may be administered as a divided dose, for example, as a divided dose administered over time. For example, in some embodiments, the dose may be administered to a subject over two or three days. An exemplary method for divided administration includes administering 25% of the dose on day 1 and the remaining 75% on day 2. In other embodiments, 33% of the dose may be administered on day 1 and the remaining 67% on day 2. In some aspects, 10% of the dose may be administered on day 1, 30% on day 2, and 60% on day 3. In some embodiments, the divided dose does not exceed three days.

[0189] In some embodiments, a dose of cells may be administered by the administration of multiple compositions or solutions, such as a first and a second, or optionally more, each containing a portion of the dose of cells. In some embodiments, multiple compositions, each containing a different population and / or subtype of cells, may be administered separately or independently, optionally within a specific time period. For example, the population or subtype of cells may each be CD8 + and CD4 + The T cells may include and / or populations rich in CD8+ and / or CD4+, respectively, such as CD4+ and / or CD8+ T cells, which include cells genetically engineered to express recombinant receptors individually. In some embodiments, a dose administration includes the administration of a first composition comprising a dose of CD8+ T cells or a dose of CD4+ T cells, and the administration of a second composition comprising a dose of the other of CD4+ T cells and CD8+ T cells.

[0190] In some embodiments, the administration of a composition or dose, for example, the administration of multiple cell compositions, includes administering the cell compositions separately. In some embodiments, the separate administrations are performed simultaneously or sequentially in any order. In some embodiments, the dose comprises a first composition and a second composition, and the first and second compositions are administered at intervals of 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours. In some embodiments, the initiation of administration of the first composition and the initiation of administration of the second composition occur at intervals of no more than 2 hours, no more than 1 hour, or no more than 30 minutes, no more than 15 minutes, no more than 10 minutes, or no more than 5 minutes. In some embodiments, the initiation and / or completion of administration of the first composition and the completion and / or initiation of administration of the second composition occur at intervals of no more than 2 hours, no more than 1 hour, or no more than 30 minutes, no more than 15 minutes, no more than 10 minutes, or no more than 5 minutes.

[0191] In some embodiments, the first composition and the second composition are mixed before administration to the subject. In some embodiments, the first composition and the second composition are mixed shortly before administration (e.g., within 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1.5 hours, 1 hour, or 0.5 hours). In some embodiments, the first composition and the second composition are mixed immediately before administration.

[0192] In some compositions, the first composition, for example, the dose of the first composition, contains CD4+ T cells. In some compositions, the first composition, for example, the dose of the first composition, contains CD8+ T cells. In some embodiments, the first composition is administered before the second composition.

[0193] In some embodiments, the dose or composition of cells comprises a defined ratio or target ratio of CD4+ cells expressing recombinant receptors versus CD8+ cells expressing recombinant receptors, and / or CD4+ cells versus CD8+ cells, where this ratio is arbitrarily about 1:1 or about 1:3 to about 3:1, e.g., about 1:1. In some aspects, the administration of a composition or dose having a target or desired ratio of different cell populations (e.g., CD4+:CD8+ ratio or CAR+CD4+:CAR+CD8+ ratio, e.g., 1:1) comprises the administration of a cell composition containing one population, followed by the administration of another cell composition containing the other population, where the administration is at the target or desired ratio or a ratio close thereto. In some aspects, the administration of a dose or composition of cells at a defined ratio results in improved expansion, persistence, and / or antitumor activity of T-cell therapy.

[0194] In some embodiments, the subject receives multiple doses of cells, e.g., two or more doses or multiple consecutive doses. In some embodiments, two doses are administered to the subject. In some embodiments, the subject receives consecutive doses, e.g., a second dose 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, multiple consecutive doses are administered after the first dose, resulting in one or more additional doses being administered after the administration of the consecutive doses. In some aspects, the number of cells administered to the subject in the additional doses is the same as or similar to the first dose and / or consecutive doses. In some embodiments, the one or more additional doses are greater than the previous doses.

[0195] In some cases, the magnitude of the initial dose and / or subsequent doses is determined based on one or more criteria, such as the subject's response to prior treatment (e.g., chemotherapy), the disease burden in the subject (e.g., tumor volume, volume, size, or degree, extent, or type of metastasis), the stage of the disease, and / or the subject's likelihood or frequency of developing toxic outcomes (e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or host immune response to administration of cells and / or recombinant receptors).

[0196] In some aspects, the period between the administration of the first dose and the administration of a subsequent dose is approximately 9 to 35 days, approximately 14 to 28 days, or 15 to 27 days. In some aspects, the administration of the subsequent dose occurs more than approximately 14 days but less than approximately 28 days after the administration of the first dose. In some aspects, the period between the first dose and the subsequent dose is approximately 21 days. In some aspects, one or more additional doses (e.g., consecutive doses) are administered after the administration of a consecutive dose. In some aspects, one or more additional consecutive doses are administered at least approximately 14 days but less than approximately 28 days after the administration of the previous dose. In some aspects, the additional dose is administered less than approximately 14 days after the previous dose, for example, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, or 13 days after the previous dose. In some embodiments, the dose is not administered less than approximately 14 days after the previous dose, and / or the dose is not administered more than approximately 28 days after the previous dose.

[0197] In some embodiments, the dose of cells (e.g., recombinant receptor-expressing cells) comprises two doses (e.g., two doses) including an initial dose of T cells and one consecutive dose of T cells, in which case one or both of the initial and second doses include the administration of a divided dose of T cells.

[0198] In some embodiments, the dose of cells is generally sufficient to be effective in reducing the disease burden.

[0199] In some embodiments, cells are administered in a desired dose, which in some aspects includes a desired dose or number of cells or cell types and / or cell types in a desired ratio. Thus, in some embodiments, the dose of cells is based on the total number of cells (or number per kg of body weight) and a desired ratio of individual populations or subtypes, e.g., CD4+ to CD8+ ratio. In some embodiments, the dose of cells is based on the desired total number of cells (or number per kg of body weight) in individual populations or individual cell types. In some embodiments, the dose is based on a combination of features such as the desired total number of cells in an individual population, a desired ratio, and a desired total number of cells.

[0200] In some embodiments, CD8 + and CD4 + A population or subtype of cells, such as T cells, is administered in or within a tolerance of a desired dose of total cells, such as a desired dose of T cells. In some embodiments, the desired dose is a desired number of cells or a desired number of cells per unit body weight of the subject to which the cells are administered, e.g., cells / kg. In some embodiments, the desired dose is a minimum number of cells or above, or a minimum number of cells per unit body weight or above. In some embodiments, of the total cells administered in the desired dose, individual populations or subtypes have a desired production ratio (CD4 + vs CD8 + A ratio (or a similar proportion) or a ratio close to it, for example, existing within a certain tolerance or error for such a ratio.

[0201] In some embodiments, cells are administered in a tolerance or within a tolerance of a desired dose of cells of one or more individual populations or subtypes, such as a desired dose of CD4+ cells and / or a desired dose of CD8+ cells. In some aspects, the desired dose is a desired number of cells of a subtype or population, or a desired number of such cells per unit body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is above the minimum number or minimum number of cells of a population or subtype, or above the minimum number or minimum number of cells of a population or subtype per unit body weight.

[0202] Therefore, in some embodiments, the dosage is based on a desired fixed dose and a desired ratio of all cells, and / or on a desired fixed dose of one or more individual subtypes or subpopulations, e.g., each of each. Therefore, in some embodiments, the dosage is based on a desired fixed dose or minimum dose of T cells and CD4 + vs CD8 + Based on the desired ratio of cells, and / or CD4 + and / or CD8 + Based on the desired fixed dose or minimum dose of cells.

[0203] In some embodiments, cells are administered within an acceptable range of a desired production ratio of multiple cell populations or subtypes, such as CD4+ and CD8+ cells or subtypes. In some embodiments, the desired ratio may be a specific ratio or a range of ratios, for example, in some embodiments, the desired ratio (e.g., CD4+) + vs CD8 +The cell ratio is 5:1 to 5:1 or approximately 5:1 to 5:1 (or greater than approximately 1:5 and less than approximately 5:1), or 1:3 to 3:1 or approximately 1:3 to 3:1 (or greater than approximately 1:3 and less than approximately 3:1), for example 2:1 to 1:5 or approximately 2:1 to 1:5 (or greater than approximately 1:5 and less than approximately 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 The ratios are 0.2:1, 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 approximately the aforementioned ratios. In some cases, the tolerance is within approximately 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, and 50% of the desired ratio, and includes any value between these ranges.

[0204] 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 all cells, T cells, or peripheral blood mononuclear cells (PBMCs) administered.

[0205] In some cases, 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., the amount, volume, or size of the tumor, or the extent, range, or type of metastasis, stage, and / or the likelihood or incidence of the subject developing toxic outcomes, e.g., CRS, macrophage activation syndrome, oncolytic syndrome, neurotoxicity, and / or the host's immune response to the cells and / or recombinant receptors being administered.

[0206] In some embodiments, the method also includes a step of administering one or more doses of cells expressing a chimeric antigen receptor (CAR) and / or lymphocyte apheresis, and / or repeating one or more steps of the method. In some embodiments, one or more further doses are the same as the initial dose. In some embodiments, one or more further doses are different from the initial dose, for example, higher than the initial dose, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 times higher or more, or lower than the initial dose, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 times lower or more. In some embodiments, the administration of one or more additional doses is determined based on the subject's response to initial treatment or any prior treatment, for example, disease burden in the subject, such as the amount, volume, size, or extent, extent, or type of tumor metastasis, stage, and / or toxic outcomes, such as CRS, macrophage activation syndrome, oncolytic syndrome, neurotoxicity, and / or the likelihood or incidence of the subject developing a host immune response to the administered cells and / or recombinant receptors.

[0207] B. Administration of compound A In some aspects of the methods, compositions, combinations, kits, or products provided herein, the combination therapy has the chemical name (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione and / or formula I: The process includes administering compound A having the structure TIFF0007837864000012.tif28128, or its enantiomer or mixture of enantiomers, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof.

[0208] In some embodiments, compound A is an enantiomer or mixture of enantiomers of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione. In some embodiments, compound A is a solvate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione. In some embodiments, compound A is the hydrate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione. In some embodiments, compound A is a pharmaceutically acceptable salt of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione. In some embodiments, compound A is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione. In some embodiments, compound A has the structure of formula I.

[0209] In some embodiments, compound A can be prepared according to the methods described in U.S. Patent No. 9,221,788 and International Publication No. 2011 / 100380, which are incorporated herein by reference in their entirety. Compound A can also be synthesized according to other available methods based on the teachings herein. In some embodiments, the pharmaceutical compositions and unit dosage forms of compound A are used according to those described in U.S. Patent No. 10,080,801. In other embodiments, compound A can be prepared or used according to U.S. Patent Application Publication No. 2014 / 0045843.

[0210] In certain embodiments, compound A is a solid. In certain embodiments, compound A is hydrated. In certain embodiments, compound A is solvated. In certain embodiments, compound A is anhydrous. In certain embodiments, compound A is nonhygroscopic.

[0211] In certain embodiments, compound A is amorphous. In certain embodiments, compound A is crystalline. In certain embodiments, solid compound A is in the crystalline form described in U.S. Patent No. 9,221,788, which is incorporated herein by reference in whole.

[0212] The solid form of compound A can be prepared according to the methods described in U.S. Patent No. 9,221,788 or any one or more of the available methods in combination.

[0213] In certain embodiments, compound A is the hydrochloride salt of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or its enantiomer or mixture of enantiomers, or a pharmaceutically acceptable solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. In certain embodiments, the hydrochloride salt is solid. In certain embodiments, the hydrochloride salt is anhydrous. In certain embodiments, the hydrochloride salt is nonhygroscopic. In certain embodiments, the hydrochloride salt is amorphous. In certain embodiments, the hydrochloride salt is crystalline.

[0214] The hydrochloride salt of compound A and its solid form can be prepared according to the methods described in U.S. Patent No. 9,221,788, or any one or more of the available methods in combination.

[0215] In some embodiments, compound A provided herein contains a single chiral center and may exist as a mixture of enantiomers, such as a racemic mixture. This disclosure encompasses the use of stereoisomerically pure forms of such compounds, as well as the use of mixtures of those forms. For example, a mixture containing equimolar or unequal amounts of enantiomers of compound A provided herein may be used in the methods and compositions disclosed herein. These isomers may be asymmetrically synthesized or resolved using standard techniques such as chiral columns or chiral resolving agents. See, for example, Jacques, J., et al, Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, SH, et al, Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions p.268 (E L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0216] It should be noted that in cases of discrepancies between the depicted structure and the name given to it, the depicted structure takes precedence. Furthermore, if the stereochemistry of a structure or part of a structure is not indicated, for example, by a thick or dashed line, the structure or part of a structure should be interpreted as encompassing all stereoisomers of the structure.

[0217] 1. Compositions and Formulations In some aspects of the combination therapy methods, compositions, combinations, kits, and uses provided herein, the combination therapy may be administered with one or more compositions, for example, a pharmaceutical composition comprising compound A.

[0218] In some embodiments, a composition, for example, a pharmaceutical composition containing compound A, may include a diluent, adjuvant, excipient, or carrier such as a vehicle, to which compound A and / or cells are administered together. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin. Such a composition generally contains a therapeutically effective amount of compound A in a purified form, along with an appropriate amount of carrier, to provide a form for appropriate administration to a patient. Such pharmaceutical carriers may be sterile liquids, such as water, as well as oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Saline solutions and aqueous solutions of dextrose and glycerol can also be used as liquid carriers, particularly for injectable solutions. A pharmaceutical composition may include one or more of the following: diluents, adjuvants, antifouling agents, binders, coatings, fillers, fragrances, colorants, lubricants, flow enhancers, preservatives, surfactants, adsorbents, emulsifiers, pharmaceutical excipients, pH buffers, or sweeteners, and combinations thereof. In some embodiments, the pharmaceutical composition may be in liquid, solid, lyophilized powder, gel, and / or combination thereof. In some aspects, the choice of carrier is determined, in part, by the specific inhibitor and / or method of administration.

[0219] Pharmacochemically acceptable carriers are generally non-toxic to the recipient at the doses and concentrations used and include, but are not limited to, the following: buffers such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, alkylparabens such as butyl or benzyl alcohol, methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); and low molecular weight (approximately 10 residues). Polypeptides (less than 100%); proteins such as serum 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 nonionic surfactants, stabilizers, and / or preservatives such as polyethylene glycol (PEG). Compositions containing compound A may also be freeze-dried.

[0220] In some embodiments, pharmaceutical compositions can be formulated for administration by any route known to those skilled in the art, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical, local, ear, inhalation, oral (e.g., sublingual), and transdermal administration or any other route. In some embodiments, other modes of administration are also intended. In some embodiments, administration is carried out by bolus injection, injection, e.g., intravenous or subcutaneous injection, intraocular injection, periorbital injection, subretinal injection, intravitreous injection, transseptal injection, subscleral injection, choroidal injection, anterior chamber injection, subconjunctival injection, subconjunctival injection, sub-Tenon's capsule injection, retrobulbar injection, peribulbar injection, or posterior parascleral delivery. In some embodiments, administration is carried out by parenteral administration, intrapulmonary administration, and intranasal administration, and, if desired for local treatment, by intralesional administration. Parenteral infusion includes intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. In some embodiments, a given dose is administered by a single bolus. In some embodiments, a given dose is administered by multiple bolus doses over a period of, for example, three days or less, or by continuous infusion.

[0221] In some embodiments, administration may be local, topical, or systemic, depending on the site of treatment. In some embodiments, local administration to the area requiring treatment may be achieved, for example, by local injection during surgery, by local application in combination with postoperative wound dressing, by injection, by catheter, by suppositories, or by implants, without limitation. In some embodiments, the composition may also be administered continuously, intermittently, or in the same composition together with other bioactive agents. In some embodiments, administration may also include a controlled release system, including a controlled release formulation and device-controlled release such as a pump. In some embodiments, administration is by oral administration.

[0222] In some embodiments, compound A is typically formulated and administered in unit-dose or multi-dose dosage forms. Each unit dose contains a predetermined amount of therapeutically active compound A sufficient to produce the desired therapeutic effect, along with the necessary pharmaceutically acceptable carrier, vehicle, or diluent. In some embodiments, unit-dose dosage forms include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions, all containing an appropriate amount of compound A. Unit-dose dosage forms may be sealed ampoules and syringes, or individually packaged tablets or capsules. Unit-dose dosage forms may be administered in fractional or multiple units. In some embodiments, multi-dose dosage forms are multiple identical unit-dose dosage forms packaged in a single container for administration in separate unit-dose dosage forms. Examples of multi-dose dosage forms include vials, bottles of tablets or capsules, or pint or gallon bottles.

[0223] 2. Medication In some embodiments, the combination therapy method provided includes the step of initiating the administration of a therapeutically effective amount of compound A before, after, during, or concurrently with the initiation of a cell therapy such as T cell therapy (e.g., CAR-expressing T cells), concurrently, nearly concurrently, sequentially, simultaneously, and / or intermittently. In some embodiments, the initiation of the administration of compound A in the combination therapy method provided is administered after or following the initiation of the administration of T cell therapy.

[0224] In some embodiments, the administration of compound A is initiated after (following) the initiation of cell therapy, such as T cell therapy (e.g., CAR-expressing T cells). In some embodiments, the administration of compound A is initiated when the peak or maximum level of T cell therapy cells is detectable in the target blood, or before.

[0225] In some cases, the initiation of administration of compound A is (i) The peak or maximum level of T-cell therapy cells is detectable in the target blood; (ii) After becoming detectable in the blood, the number of T-cells detected in the blood is undetectable or decreased, or optionally decreased compared to a preceding point in time after administration of T-cell therapy; (iii) The number of T-cells detectable in the blood has decreased by 1.5 times, 2.0 times, 3.0 times, 4.0 times, 5.0 times, 10 times, or more than the peak or maximum number of T-cells detectable in the subject's blood after the start of T-cell therapy administration; (iv) At the time after the peak or maximum level of T-cell therapy cells becomes detectable in the subject's blood, the number of detectable cells or cells derived from detectable cells 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 has shown disease progression after treatment with T-cell therapy and / or has relapsed after remission; and / or (iv) The subject shows an increased tumor burden compared to the tumor burden before or after cell administration and before the start of administration of compound A. It is performed one week prior to or within one week prior to, for example, within one, two, or three days. In certain situations, the methods provided are performed to enhance, increase, or intensify T cell therapy in a subject in order to improve the response to T cell therapy, for example, the presence of T cells and / or reduction of tumor burden.

[0226] In some embodiments, the administration of compound A is initiated after (following) the initiation of cell therapy, such as T cell therapy (e.g., CAR-expressing T cells). In some embodiments, the administration of compound A is initiated early, after (following) the initiation of T cell therapy, and on the same day as the initiation of T cell therapy (i.e., as early as day 0 after the initiation of T cell therapy). In some embodiments, the administration of compound A is initiated from day 0 or approximately day 0 to day 21 or approximately 21 days after the initiation of T cell therapy. In some embodiments, the administration of compound A is initiated approximately 7 to approximately 14 days after the initiation of T cell therapy. In some embodiments, the administration of compound A is initiated from day 0 or approximately day 0 to day 14 or approximately 14 days after the initiation of T cell therapy. In some embodiments, the administration of compound A is initiated on day 0 of the initiation of T cell therapy (i.e., the administration of compound A is initiated on the same day as the initiation of T cell therapy).

[0227] In some embodiments, administration of compound A is initiated 1 day or about 1 day to 21 days or about 21 days after the start of T-cell therapy. In some embodiments, administration of compound A is initiated 1 day or about 1 day to 15 days or about 15 days after the start of T-cell therapy. In some embodiments, administration of compound A is initiated about 8 to about 15 days after the start of T-cell therapy. In some embodiments, the administration of compound A is initiated 1 day or approximately 1 day, 2 days or approximately 2 days, 3 days or approximately 3 days, 4 days or approximately 4 days, 5 days or approximately 5 days, 6 days or approximately 6 days, 7 days or approximately 7 days, 8 days or approximately 8 days, 9 days or approximately 9 days, 10 days or approximately 10 days, 11 days or approximately 11 days, 12 days or approximately 12 days, 13 days or approximately 13 days, 14 days or approximately 14 days, 15 days or approximately 15 days, 16 days or approximately 16 days, 17 days or approximately 17 days, 18 days or approximately 18 days, 19 days or approximately 19 days, or 20 days or approximately 20 days after the start of T-cell therapy. In some embodiments, the administration of compound A is initiated 15 days or approximately 15 days after the start of T-cell therapy. In some embodiments, administration of compound A is initiated 14 days or approximately 14 days after the start of T-cell therapy. In some embodiments, administration of compound A is initiated 8 days or approximately 8 days after the start of T-cell therapy. In some embodiments, administration of compound A is initiated 7 days or approximately 7 days after the start of T-cell therapy. In other embodiments, administration of compound A is initiated 1 day or approximately 1 day after the start of T-cell therapy.

[0228] Therefore, with respect to combination therapies in which T-cell therapy is administered on day 1 of the combination therapy, in some embodiments, administration of compound A is initiated on day 2 or approximately day 22 or approximately day 22 of the combination therapy. In some embodiments, administration of compound A is initiated on day 2 or approximately day 2 or approximately day 16 or approximately day 16 of the combination therapy. In some embodiments, administration of compound A is initiated on day 9 or approximately day 9 or approximately day 16 or approximately day 16 of the combination therapy. In some embodiments, administration of compound A is initiated on day 8 or approximately day 8 or approximately day 15 or approximately day 15 of the combination therapy. In some embodiments, administration of compound A is initiated on day 2 or approximately day 2, day 3 or approximately day 3, day 4 or approximately day 4, day 5 or approximately day 5, day 6 or approximately day 6, day 7 or approximately day 7, day 8 or approximately day 8, day 9 or approximately day 9, day 10 or approximately day 10, day 11 or approximately day 11, day 12 or approximately day 12, day 13 or approximately day 13, day 14 or approximately day 14, day 15 or approximately day 15, day 16 or approximately day 16, day 17 or approximately day 17, day 18 or approximately day 18, day 19 or approximately day 19, day 20 or approximately day 20, or day 21 or approximately day 21. In some embodiments, administration of compound A is initiated on day 16 or approximately day 16 of the combination therapy. In some embodiments, administration of compound A is initiated on day 15 or approximately day 15 of the combination therapy. In some embodiments, administration of compound A is initiated on day 9 or approximately day 9 of the combination therapy. In some embodiments, administration of compound A is initiated on day 8 or approximately day 8 of the combination therapy. In some embodiments, administration of compound A is initiated on day 2 or approximately day 2 of the combination therapy.

[0229] In other embodiments, administration of compound A is initiated on day 1 or approximately day 1 of the combination therapy.

[0230] In some embodiments, at the time of initial administration of compound A, and / or at any subsequent point after the start of administration, the subject does not exhibit signs or symptoms of severe toxicity, such as severe cytokine release syndrome (CRS), or severe toxicity. In some embodiments, administration of compound A is performed 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., long-term grade 3 CRS or grade 4 or 5 CRS. In some embodiments, administration of compound A is performed 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., long-term grade 3 neurotoxicity or grade 4 or 5 neurotoxicity. In some aspects, between the start of T-cell therapy administration and the administration of compound A, the subject did not exhibit severe CRS, and / or grade 3 or higher CRS, e.g., long-term grade 3 CRS or grade 4 or 5 CRS. In some cases, subjects did not exhibit severe neurotoxicity and / or grade 3 or higher neurotoxicity, such as long-term grade 3 neurotoxicity or grade 4 or 5 neurotoxicity, between the start of T-cell therapy and the administration of compound A.

[0231] In some embodiments, the daily dose of compound A administered is 0.1 mg to 1.0 mg or approximately 0.1 mg to 1.0 mg. In some embodiments, the daily dose of compound A administered is approximately 0.1 mg to 1.0 mg, approximately 0.2 mg to 1.0 mg, approximately 0.3 mg to 1.0 mg, approximately 0.4 mg to 1.0 mg, approximately 0.5 mg to 1.0 mg, approximately 0.6 mg to 1.0 mg, approximately 0.7 mg to 1.0 mg, approximately 0.8 mg to 1.0 mg, approximately 0.9 mg to 1.0 mg, approximately 0.1 mg to 0.80 mg, approximately 0.2 mg to 0.80 mg, approximately 0.3 mg to 0.80 mg, approximately 0.4 mg to 0. The amounts are 0.80 mg, approximately 0.5 mg to approximately 0.80 mg, approximately 0.6 mg to approximately 0.80 mg, approximately 0.7 mg to approximately 0.80 mg, approximately 0.1 mg to approximately 0.60 mg, approximately 0.2 mg to approximately 0.60 mg, approximately 0.3 mg to approximately 0.60 mg, approximately 0.4 mg to approximately 0.60 mg, approximately 0.5 mg to approximately 0.60 mg, approximately 0.1 mg to approximately 0.40 mg, approximately 0.2 mg to approximately 0.40 mg, approximately 0.3 mg to approximately 0.40 mg, approximately 0.1 mg to approximately 0.20 mg, or approximately 0.1 mg to approximately 0.3 mg. In some embodiments, compound A is administered in amounts of approximately 0.3 mg to approximately 0.6 mg / day.

[0232] In some embodiments, the daily dose of compound A administered is approximately 0.1 mg or at least approximately 0.1 mg, or 0.1 mg or at least 0.1 mg. In some embodiments, the daily dose of compound A administered is approximately 0.2 mg or at least approximately 0.2 mg, or 0.2 mg or at least 0.2 mg. In some embodiments, the daily dose of compound A administered is approximately 0.3 mg or at least approximately 0.3 mg, or 0.3 mg or at least 0.3 mg. In some embodiments, the daily dose of compound A administered is approximately 0.4 mg or at least approximately 0.4 mg, or 0.4 mg or at least 0.4 mg. In some embodiments, the daily dose of compound A administered is approximately 0.5 mg or at least approximately 0.5 mg, or 0.5 mg or at least 0.5 mg. In some of any such embodiments, the daily dose of compound A administered is approximately 5.0 mg or less. In some of these embodiments, the daily dose of compound A administered is approximately 1.0 mg or less. In some embodiments, the daily dose of compound A administered is approximately 0.8 mg or less. In some embodiments, the daily dose of compound A administered is approximately 0.6 mg or less. In some embodiments, the daily dose of compound A administered is approximately 0.5 mg or less.

[0233] In some embodiments, the daily dose of compound A is 0.5 mg or approximately 0.5 mg. In some embodiments, the daily dose of compound A is 0.45 mg or approximately 0.45 mg. In some embodiments, the daily dose of compound A is 0.30 mg or approximately 0.30 mg. In some embodiments, the daily dose of compound A is 0.60 mg or approximately 0.60 mg.

[0234] In some embodiments, compound A reaches the maximum concentration of compound A in the blood (C maxThe amount that achieves ) for each week of the cycling regimen, or for at least one week of the cycling regimen, for example, 1nM to 20nM or about 1nM to about 20nM, 1nM to 15nM or about 1nM to about 15nM, 1nM to 12nM or about 1nM to about 12nM, 1nM to 10nM or about 1nM to about 10nM, 1nM to 5nM or about 1nM to about 5nM, 1nM to 2.5nM or about 1nM to about 2.5nM, 2.5nM to 20nM or about 2.5nM to about 20nM, 2.5nM to 15nM or about 2.5nM to about 15nM, 2.5nM to 12nM Alternatively, it is administered in the range of approximately 2.5 nM to approximately 12 nM, 2.5 nM to 10 nM, or approximately 2.5 nM to approximately 10 nM, 2.5 nM to 5 nM, or approximately 2.5 nM to approximately 5 nM, 5 nM to 20 nM, or approximately 5 nM to approximately 20 nM, 5 nM to 15 nM, or approximately 5 nM to approximately 15 nM, 5 nM to 12 nM, or approximately 5 nM to approximately 12 nM, 5 nM to 10 nM, or approximately 5 nM to approximately 10 nM, 10 nM to 20 nM, or approximately 10 nM to approximately 20 nM, 10 nM to 15 nM, or approximately 10 nM to approximately 15 nM, and 15 nM to 20 nM or approximately 15 nM to approximately 20 nM (each including the values ​​at both ends). In some embodiments, compound A is C max It is administered in a dose that maintains the state for at least approximately 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 16 hours, or 24 hours.

[0235] In some embodiments, compound A is released into the bloodstream. max The amount that achieves this is administered, for example, at about 1 nM or at least about 1 nM for each week of the cycling regimen, or for at least one week of the cycling regimen. In some embodiments, compound A is administered in the blood at a concentration of C max The amount administered is such that the desired result is achieved, for example, for each week of the cycling regimen, or for at least one week of the cycling regimen, at a dose of about 10 nM or at least about 10 nM. In some embodiments, compound A is administered in the blood at a concentration of C maxThe amount that achieves this is administered, for example, at about 5 nM or at least about 5 nM for each week of the cycling regimen, or for at least one week of the cycling regimen. In some embodiments, compound A is administered in the blood at a concentration of C max The amount administered is, for example, about 2.5 nM or at least about 2.5 nM for each week of the cycling regimen, or for at least one week of the cycling regimen, in an amount that achieves the desired result. In some embodiments, compound A is C max It is administered in a dose that maintains the effect for at least approximately 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 16 hours, or 24 hours.

[0236] In some embodiments, compound A is administered in a cycling regimen (also referred to herein as cycling therapy) that involves repeated administration of the compound over a specific period or duration. In some embodiments, the amount of compound A per dose or per day administered is 1.0 mg or less (e.g., 1.0 mg, 0.9 mg, 0.8 mg, 0.7 mg, 0.6 mg, 0.5 mg or less). In some embodiments, the amount of compound A per dose or per day administered is 0.6 mg or about 0.6 mg, 0.5 mg or about 0.5 mg, 0.45 mg or about 0.45 mg, 0.40 mg or about 0.40 mg, 0.30 mg or about 0.30 mg, 0.2 mg or about 0.2 mg. In some embodiments, the amount of compound A per dose or per day administered is approximately 0.30 mg to approximately 0.60 mg (e.g., 0.30 mg or approximately 0.30 mg, 0.45 mg or approximately 0.45 mg, or 0.60 mg or approximately 0.60 mg).

[0237] In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) within 21 days after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 21 days, 20 days, 19 days, 18 days, 17 days, 16 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 15 ± 3 days after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 14 days after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 8 ± 3 days after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 7 days after the administration of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is initiated (or started) 1 or 2 days after the administration of T-cell therapy.

[0238] Therefore, with respect to combination therapies in which T-cell therapy is administered on day 1 of the combination therapy, in some embodiments, the administration of compound A in the cycling regimen is initiated (or is initiated) on day 22 of the combination therapy or earlier. In some embodiments, the administration of compound A in the cycling regimen is initiated (or is initiated) on day 22, or day 21, or day 20, or day 19, or day 18, or day 17, or day 16, or day 15, or day 14, or day 13, or day 12, or day 11, or day 10, or day 9, or day 8, or day 7, or day 6, or day 5, or day 4, or day 3, or day 2 of the combination therapy. In some embodiments, the administration of compound A in the cycling regimen is initiated (or is initiated) on day 16±3 of the combination therapy. In some embodiments, the administration of compound A in the cycling regimen is initiated (or is initiated) on day 15 of the combination therapy. In some embodiments, administration of compound A in the cycling regimen is initiated (or initiated) on day 9 ± 3 of the combination therapy. In some embodiments, administration of compound A in the cycling regimen is initiated (or initiated) on day 8 of the combination therapy. In some embodiments, administration of compound A in the cycling regimen is initiated (or initiated) on day 2 or 3 of the combination therapy.

[0239] In other embodiments, administration of compound A in the cycling regimen is initiated (or is initiated) on day 1 of the combination therapy.

[0240] In some embodiments, a cycling regimen for the administration of compound A includes a first administration period in which the compound is administered daily for up to three consecutive weeks, a rest period in which the compound is not administered, beginning at the end of the first administration period, and a second administration period comprising a four-week cycle in which the compound is administered daily for three consecutive weeks over a four-week period.

[0241] In some embodiments, the first administration period begins (or is initiated) within 21 days after the administration of T-cell therapy. In some embodiments, the first administration period begins 21 days, or 20 days, or 19 days, or 18 days, or 17 days, or 16 days, or 15 days, or 14 days, or 13 days, or 12 days, or 11 days, or 10 days, or 9 days, or 8 days, or 7 days, or 6 days, or 5 days, or 4 days, or 3 days, or 2 days, or 1 day after the administration of T-cell therapy. In some embodiments, the first administration period begins 15 ± 3 days after the administration of T-cell therapy. In some embodiments, the first administration period begins 14 days after the administration of T-cell therapy. In some embodiments, the first administration period begins 8 ± 3 days after the administration of T-cell therapy. In some embodiments, the first administration period begins 7 days after the administration of T-cell therapy. In some embodiments, the first administration period begins 1 or 2 days after the administration of T-cell therapy. In some embodiments, the compound is administered at approximately 0.1 mg / day to approximately 1.0 mg / day during the first administration period. In some embodiments, the compound is administered at approximately 0.45 mg / day during the first administration period. In some embodiments, the compound is administered at approximately 0.3 mg / day during the first administration period. In some embodiments, the compound is administered at approximately 0.60 mg / day during the first administration period.

[0242] Therefore, with respect to combination therapies in which T-cell therapy is administered on day 1 of the combination therapy, in some embodiments, the first administration period begins (or is initiated) on day 22 of the combination therapy or earlier. In some embodiments, the first administration period begins (or is initiated) on day 22, or day 21, or day 20, or day 19, or day 18, or day 17, or day 16, or day 15, or day 14, or day 13, or day 12, or day 11, or day 10, or day 9, or day 8, or day 7, or day 6, or day 5, or day 4, or day 3, or day 2 of the combination therapy. In some embodiments, the first administration period begins (or is initiated) on day 16±3 of the combination therapy. In some embodiments, the first administration period begins (or is initiated) on day 15 of the combination therapy. In some embodiments, the first administration period begins (or is initiated) on day 9±3 of the combination therapy. In some embodiments, the first administration period begins (or is initiated) on day 8 of the combination therapy. In some embodiments, the first administration period begins (or is initiated) on day 2 or 3 of the combination therapy.

[0243] In other embodiments, the first administration period begins on day 1 of the combination therapy.

[0244] In some embodiments, compound A is administered daily for 1 to 21 days or approximately 1 to 21 days, 1 to 19 days or approximately 1 to 19 days, 1 to 17 days or approximately 1 to 17 days, 1 to 15 days or approximately 1 to 15 days, 1 to 13 days or approximately 1 to 13 days, 1 to 11 days or approximately 1 to 11 days, 1 to 9 days or approximately 1 to 9 days, 1 to 7 days or approximately 1 to 7 days, 1 to 5 days or approximately 1 to 5 days, or 1 to 3 days or approximately 1 to 3 days (including the values ​​at both ends). In some embodiments, compound A is administered daily for 1 to 21 days or approximately 1 to 21 days (including the values ​​at both ends) during the first administration period. In some embodiments, compound A is administered daily for 1 to 14 days or approximately 1 to 14 days (including the values ​​at both ends) during the first administration period. In some embodiments, compound A is administered daily for 7 to 21 days or approximately 7 to 21 days (including the values ​​at both ends) during the first administration period. In some embodiments, compound A is administered daily for 21 days or approximately 21 days during the first administration period. In some embodiments, compound A is administered daily for 14 days or approximately 14 days during the first administration period. In some embodiments, compound A is administered daily for 7 days or approximately 7 days during the first administration period.

[0245] It is understood that the first administration ends at the start of the rest period. In some embodiments, the rest period begins approximately 22 days after administration of T-cell therapy. In some embodiments, the rest period begins approximately 19, 20, 21, 22, 23, or 24 days after administration of T-cell therapy. In some embodiments, the rest period begins 22 days after administration of T-cell therapy. In some embodiments, the rest period begins 21 days after administration of T-cell therapy. In some embodiments, the rest period is approximately one week. In some embodiments, the rest period is 5, 6, 7, 8, or 9 days. In some embodiments, the rest period is 7 days. In some embodiments, the rest period continues until the absolute neutrophil count (ANC) is the same as or approximately the same as the level measured before administration of T-cell therapy. In some embodiments, the rest period is 7 days. In some embodiments, the rest period continues until the absolute neutrophil count (ANC) is the same as or nearly the same as the level measured before the first administration period. In some embodiments, the rest period continues until the subject's B cell count level recovers to the same as or nearly the same as the level measured before T cell therapy was administered. In some embodiments, the rest period continues until the subject's B cell count level recovers to the same as or nearly the same as the level measured before the first administration period.

[0246] The rest period is understood to end at the start of the second administration period. In some embodiments, the second administration period begins approximately 29 days after the administration of T-cell therapy. In some embodiments, the second administration period begins 27, 28, 29, 30, or 31 days after the administration of T-cell therapy. In some embodiments, the second administration period begins 29 days after the administration of T-cell therapy. In some embodiments, the second administration period begins 28 days after the administration of T-cell therapy. In some embodiments, the compound is administered at approximately 0.1 mg / day to approximately 1.0 mg / day during the second administration period. In some embodiments, the compound is administered at approximately 0.45 mg / day during the second administration period. In some embodiments, the compound is administered at approximately 0.3 mg / day during the second administration period. In some embodiments, the compound is administered at approximately 0.60 mg / day during the second administration period. In some embodiments, the second administration period comprises a 4-week cycle in which the compound is administered daily for three consecutive weeks over a 4-week period. In some embodiments, within each 4-week cycle, the compound is not administered for one week after three consecutive weeks in which the compound is administered daily. In some embodiments, the second administration period comprises more than one 4-week cycle. In some embodiments, the second administration period comprises two 4-week cycles, or three 4-week cycles, or four 4-week cycles, five 4-week cycles, or six 4-week cycles, or seven 4-week cycles, eight 4-week cycles, or nine 4-week cycles, or ten 4-week cycles, eleven 4-week cycles, or twelve 4-week cycles, or thirteen 4-week cycles, fourteen 4-week cycles, or fifteen 4-week cycles, or sixteen 4-week cycles. In some embodiments, the second administration comprises 2 to 11 or approximately 2 to 11 four-week cycles, 2 to 9 or approximately 2 to 9 four-week cycles, 2 to 7 or approximately 2 to 7 four-week cycles, 2 to 5 or approximately 2 to 5 four-week cycles, or 2 to 4 or approximately 2 to 4 four-week cycles (including the values ​​at both ends). In some embodiments, the second administration comprises 2 to 11 or approximately 2 to 11 four-week cycles (including the values ​​at both ends). In some embodiments, the second administration period comprises two four-week cycles.In some embodiments, the second administration period comprises five 4-week cycles. In some embodiments, the second administration period comprises eleven 4-week cycles. In some embodiments, the second administration period consists of two 4-week cycles. In some embodiments, administration of compound A is initiated (or initiated) within 21 days after administration of T-cell therapy and is carried out in a cycling regimen comprising: a first administration period in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for up to three consecutive weeks; a rest period of at least one week in which the compound is not administered, beginning at the end of the first administration period; and a second administration period comprising four-week cycles in which the compound is administered daily at approximately 0.1 mg to approximately 1.0 mg / day for three consecutive weeks over a period of four weeks. In some embodiments, the compound is administered at approximately 0.30 mg, 0.45 mg, or 0.60 mg / day during the first and second administration periods. In some embodiments, the administration of compound A includes a first administration period that begins on day 15 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ends after day 21. In some embodiments, the administration of compound A includes a first administration period that begins on day 8 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ends after day 21. In some embodiments, the administration of compound A includes a first administration period that begins on day 1 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ends after day 21. In some embodiments, the rest period begins on day 22 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ends after day 28. In some embodiments, the second administration period begins on day 29 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1). In some embodiments, the second administration period begins 29 days after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ends after 180 days. In certain embodiments, the administration of compound A includes a first administration period beginning 15 days after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after 21 days, a rest period beginning 22 days after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after 28 days, and a second administration period beginning 29 days after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1).In some embodiments, the administration of compound A includes a first administration period starting on day 8 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 21, a rest period starting on day 22 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 28, and a second administration period starting on day 29 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1). In other embodiments, the administration of compound A includes a first administration period starting on day 8 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 21, a rest period starting on day 22 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 28, and a second administration period starting on day 29 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1). In other embodiments, the administration of compound A includes a first administration period beginning on day 1 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 21, a rest period beginning on day 22 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1) and ending after day 28, and a second administration period beginning on day 29 after the administration of T-cell therapy (with respect to T-cell therapy administered on day 1).

[0247] In some embodiments, a cycling regimen for administering compound A is performed for a period following the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, extends for a period exceeding one week after the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, extends for a period of about one month or at least about one month after the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, extends for a period of about two months or at least about two months after the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, extends for a period of about three months or at least about three months after the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, extends for a period of about four months or at least about four months after the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of about 5 months or at least about 5 months after the start of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of about 6 months or at least about 6 months after the start of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of about 8 months or at least about 8 months after the start of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of about 10 months or at least about 10 months after the start of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of about 12 months or at least about 12 months after the start of T-cell therapy.

[0248] In some embodiments, the administration of compound A, for example in a cycling regimen, is for a period of at least 3 months. In some embodiments, the administration of compound A, for example in a cycling regimen, is for 90 days or about 90 days, 100 days or about 100 days, 105 days or about 105 days, 110 days or about 110 days, 115 days or about 115 days, 120 days or about 120 days, 125 days or about 125 days, 130 days or about 130 days, 135 days or about 135 days, 140 days or Over a period of approximately 140 days, 145 days or approximately 145 days, 150 days or approximately 150 days, 155 days or approximately 155 days, 160 days or approximately 160 days, 165 days or approximately 165 days, 170 days or approximately 170 days, 175 days or approximately 175 days, 180 days or approximately 180 days, 185 days or approximately 185 days, 190 days or approximately 190 days, 195 days or approximately 195 days, 200 days or approximately 200 days or longer.

[0249] In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of 90 days or approximately 90 days or 3 months or approximately 3 months after the start of administration of T-cell therapy (e.g., CAR T-cell therapy). In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of 120 days or approximately 120 days or 4 months or approximately 4 months after the start of administration of T-cell therapy (e.g., CAR T-cell therapy). In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of 150 days or approximately 150 days or 5 months or approximately 5 months after the start of administration of T-cell therapy (e.g., CAR T-cell therapy). In some embodiments, the administration of compound A, for example in a cycling regimen, is over a period of 180 days or approximately 180 days or 6 months or approximately 6 months after the start of administration of T-cell therapy (e.g., CAR T-cell therapy).

[0250] In some embodiments, the cycling regimen is terminated 3 months or approximately 3 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 4 months or approximately 4 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 5 months or approximately 5 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 6 months or approximately 6 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 8 months or approximately 8 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 10 months or approximately 10 months after the start of T-cell therapy administration. In some embodiments, the cycling regimen is terminated 12 months or approximately 12 months after the start of T-cell therapy administration.

[0251] In some embodiments, the cycling regimen ends 84 days or approximately 84 days after the start of T-cell therapy administration. In some embodiments, the second administration period ends 84 days or approximately 84 days after the start of T-cell therapy administration. Therefore, with respect to a combination therapy in which T-cell therapy is administered on day 1 of the combination therapy, the second administration period ends on day 85 of the combination therapy.

[0252] In some embodiments, the cycling regimen includes a first administration period starting 14 days after administration of T-cell therapy and ending 20 days after administration of T-cell therapy, a rest period starting 21 days after administration of T-cell therapy and ending 27 days after administration of T-cell therapy, and a second administration period starting 28 days after administration of T-cell therapy and ending 84 days after administration of T-cell therapy. In some embodiments, the cycling regimen includes a first administration period starting 7 days after administration of T-cell therapy and ending 20 days after administration of T-cell therapy, a rest period starting 21 days after administration of T-cell therapy and ending 27 days after administration of T-cell therapy, and a second administration period starting 28 days after administration of T-cell therapy and ending 84 days after administration of T-cell therapy. In some embodiments, the cycling regimen includes a first administration period beginning on the same day as the administration of T-cell therapy and ending 20 days after the administration of T-cell therapy, a rest period beginning 21 days after the administration of T-cell therapy and ending 27 days after the administration of T-cell therapy, and a second administration period beginning 28 days after the administration of T-cell therapy and ending 84 days after the administration of T-cell therapy.

[0253] Therefore, with respect to combination therapy in which T-cell therapy is administered on day 1 of the combination therapy, in some embodiments, the cycling regimen includes a first administration period beginning on day 15 of the combination therapy and ending on day 21, a rest period beginning on day 22 of the combination therapy, and a second administration period beginning on day 29 of the combination therapy and ending on day 85. In some embodiments, the cycling regimen includes a first administration period beginning on day 8 of the combination therapy and ending on day 21, a rest period beginning on day 22 of the combination therapy, and a second administration period beginning on day 29 of the combination therapy and ending on day 85. In some embodiments, the cycling regimen includes a first administration period beginning on day 1 of the combination therapy and ending on day 21, a rest period beginning on day 22 of the combination therapy, and a second administration period beginning on day 29 of the combination therapy and ending on day 85.

[0254] In some embodiments, the administration of compound A, for example in a cycling regimen, is terminated or stopped at the end of a period following the initiation of T-cell therapy (e.g., 3, 4, 5, or 6 months, or approximately 3, 4, 5, or 6 months) if the subject achieves a complete response (CR) after treatment before, at, or approximately 6 months, or if the cancer (e.g., B-cell malignancy) progresses after treatment or relapses after remission. In some embodiments, the administration of compound A, for example in a cycling regimen, is terminated or stopped 3 months after the initiation of T-cell therapy (e.g., CAR T-cell therapy) if the subject achieves a complete response (CR) after treatment before, at, or approximately 3 months after the initiation of T-cell therapy, or if the cancer (e.g., B-cell malignancy) progresses after treatment or relapses after remission. In some embodiments, the period is a fixed duration during which the administration of compound A, for example, in a cycling regimen, continues even if the subject achieves a complete response (CR) 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-free.

[0255] In some embodiments, administration of compound A, for example in a cycling regimen, is continued after the end of the period if the subject shows a partial response (PR) or stable disease (SD) after treatment, for longer than 3, 4, 5, or 6 months after the initiation of T-cell therapy (e.g., CAR T-cell therapy), or for approximately 3, 4, 5, or 6 months. In some embodiments, administration of compound A, for example in a cycling regimen, is continued beyond 3 months after the initiation of T-cell therapy (e.g., CAR T-cell therapy). In some embodiments, administration of compound A, for example in a cycling regimen, is continued beyond 6 months after the initiation of T-cell therapy (e.g., CAR T-cell therapy). In some embodiments, for subjects who show a PR or SD at the end of the initial period, administration of compound A, for example in a cycling regimen, is continued until the subject achieves a complete response (CR) after treatment, or until the cancer (e.g., NHL, DLBCL, or other B-cell malignancies) progresses after treatment or relapses after remission.

[0256] In some embodiments, the administration of compound A is carried out in a cycling regimen that includes administering compound A in amounts of approximately 1.0 mg or less per day (e.g., 0.1–1.0 mg, 0.30 mg, 0.45 mg, or 0.60 mg). In some embodiments, at the time of administration of compound A, the subject does not exhibit severe toxicity after administration of T cell therapy (e.g., CAR T cells). In some embodiments, the B cell malignancy is a highly malignant NHL such as relapsed / refractory NHL or DLBCL. In some embodiments, the cell therapy, such as CAR-expressing T cells, includes a chimeric antigen receptor that specifically binds to the B cell antigen. In some embodiments, the B cell antigen is CD19.

[0257] In some embodiments, the administration of compound A is carried out in a cycling regimen that includes administering an effective amount of the compound over a period of 3 months or approximately 3 months or more than 3 months, 4 months or approximately 4 months or more than 4 months, 5 months or approximately 5 months or more than 5 months, or 6 months or approximately 6 months or more than 6 months after the start of administration of cell therapy (e.g., T-cell therapy). In some embodiments, the period is 3 months or approximately 3 months, 4 months or approximately 4 months, 5 months or approximately 5 months, or 6 months or approximately 6 months. In some embodiments, at the time of administration of compound A, the subject does not exhibit severe toxicity after administration of cell therapy. In some embodiments, the administration of compound A, e.g., the administration of compound A in a cycling regimen, is terminated or stopped if the subject achieves a complete response (CR) after treatment, or if the cancer, e.g., B-cell malignancy, progresses after treatment or relapses after remission, before or near the end of the period. In some embodiments, the administration of compound A, for example in a cycling regimen, is continued throughout the period even if the subject achieves a complete response (CR) before the end of that period. In some embodiments, the administration of compound A, for example in a cycling regimen, is continued after the end of the initial period if the subject shows a partial response (PR) or stable disease (SD) after treatment following the initiation of T-cell therapy. In some embodiments, the administration of compound A, for example in a cycling regimen, is repeated until the subject achieves a complete response (CR) after treatment, or until the cancer, for example, progresses after treatment, or relapses after remission. In some embodiments, the B-cell malignancy is a highly malignant NHL such as relapsed / refractory NHL or DLBCL. In some embodiments, the T-cell therapy, such as CAR-expressing T cells, includes a chimeric antigen receptor that specifically binds to the B-cell antigen. In some embodiments, the B-cell antigen is CD19.

[0258] In some embodiments, the administration of compound A is carried out in a cycling regimen that includes administering compound A at a rate of approximately 3 mg or less per day (e.g., 1-3 mg, 1 mg, 2 mg, or 3 mg) on ​​5 days or less per week (e.g., 3, 4, or 5 days) for a period of approximately 3 months or more than 3 months after the initiation of T-cell therapy (e.g., CAR T-cell therapy) (e.g., a period of 3 months or approximately 3 months, 4 months or approximately 4 months, 5 months or approximately 5 months, or 6 months or approximately 6 months). In some embodiments, at the time of administration of compound A, the subject does not exhibit severe toxicity after the administration of cell therapy. In some embodiments, the B-cell malignancy is a relapsed / refractory, highly malignant NHL or NHL such as DLBCL. In some embodiments, the administration of compound A, for example in a cycling regimen, is terminated or stopped 6 months or approximately 6 months after the start of T-cell therapy if the subject achieves a complete response (CR) after treatment more than 6 months or approximately 6 months prior to treatment, or if the cancer, such as B-cell malignancy, progresses or relapses after remission after treatment. In some embodiments, the cycling regimen is continued for the entire period even if the subject achieves a complete response (CR) before the end of the period. In some embodiments, the administration of compound A, for example in a cycling regimen, is continued beyond the end of the period, for example beyond 6 months after the start of cell therapy, if the subject shows a partial response (PR) or stable disease (SD) after treatment at 6 months or approximately 6 months. In some embodiments, the administration of compound A, for example in a cycling regimen, is continued until the subject achieves a complete response (CR) after treatment, or until the cancer, such as B-cell malignancy, progresses or relapses after remission after treatment. In some embodiments, cell therapies, such as CAR-expressing T cells, include chimeric antigen receptors that specifically bind to B cell antigens. In some embodiments, the B cell antigen is CD19.

[0259] In some embodiments, the administration of compound A is carried out in a cycling regimen, which involves administering compound A at a dose of approximately 1 mg to approximately 3 mg (e.g., 1 mg, 2 mg, or 3 mg) per day for a period of 6 months or approximately 6 months or more after the initiation of cell therapy (e.g., T-cell therapy). In some embodiments, the administration of compound A, e.g., in a cycling regimen, is initiated approximately 14 to approximately 35 days (e.g., approximately 21 to approximately 35 days, e.g., 28 days or approximately 28 days) after the initiation of cell therapy. In some embodiments, at the time of administration of compound A, the subject does not exhibit severe toxicity after the administration of cell therapy. In some embodiments, the administration of compound A, e.g., in a cycling regimen, is discontinued 6 months or approximately 6 months after the initiation of cell therapy if the subject achieves a complete response (CR) after treatment at 6 months or approximately 6 months, or if the cancer, e.g., B-cell malignancy, progresses after treatment or relapses after remission. In some embodiments, administration of compound A, for example in a cycling regimen, is continued during the period even if the subject achieves a complete response (CR) earlier than or approximately earlier than 6 months. In some embodiments, administration of compound A, for example in a cycling regimen, is continued beyond 6 months after the start of T-cell therapy if the subject shows a partial response (PR) or stable disease (SD) after 6 months or approximately 6 months. In some embodiments, administration of compound A, for example in a cycling regimen, is continued until the subject achieves a complete response (CR) after therapy, or until the B-cell malignancy progresses after therapy or relapses after remission. In some embodiments, the B-cell malignancy is a highly malignant NHL such as relapsed / refractory NHL or DLBCL. In some embodiments, the cell therapy, such as CAR-expressing T cells, includes a chimeric antigen receptor that specifically binds to the B-cell antigen. In some embodiments, the B-cell antigen is CD19.

[0260] In some cases, the cycling regimen may be interrupted at any point and / or once or more times. In some cases, the cycling regimen is interrupted or modified if the 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. In some embodiments, after the subject develops one or more adverse events, dose-limiting toxicity (DLT), neutropenia or febrile neutropenia, thrombocytopenia, cytokine release syndrome (CRS), and / or neurotoxicity (NT), the amount of compound A per dose or per day is changed on specific days of the week.

[0261] II. Cell therapy and cell manipulation In some embodiments, a cell therapy (e.g., T-cell therapy) for use according to a provided combination therapy method includes the step of administering engineered cells expressing recombinant receptors designed to recognize and / or specifically bind to antigens associated with a disease or condition, such as cancer, e.g., B-cell malignancies. In some embodiments, binding to the antigen results in a response, such as an immune response to such antigen. In some embodiments, the cells contain or are engineered to contain engineered antigen receptors, such as engineered receptors or recombinant receptors, e.g., chimeric antigen receptors (CARs). Recombinant receptors such as CARs generally contain, in some aspects, an extracellular antigen (or ligand) binding domain linked to one or more intracellular signaling components via a linker and / or transmembrane domain(s). In some aspects, the engineered cells are provided as 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. In some embodiments, the methods are any of those described in Section I.

[0262] In some embodiments, cells contain one or more nucleic acids introduced by genetic engineering, thereby expressing recombinant or genetically engineered products of such nucleic acids. In some embodiments, gene transfer is achieved by first stimulating cells, such as by combining them with stimuli that induce responses such as proliferation, survival, and / or activation, as measured by the expression of cytokines or activation markers, and then transducing the activated cells to expand to a number sufficient for clinical application under culture conditions.

[0263] A. Chimeric antigen receptor For example, as described in any of the methods and uses provided, in some embodiments of the methods and uses, engineered cells such as T cells express a chimeric receptor, such as a chimeric antigen receptor (CAR), which comprises one or more domains that combine an intracellular signaling domain with a ligand-binding domain (e.g., an antibody or antibody fragment) that provides specificity to a desired antigen (e.g., a tumor antigen). In some embodiments, the intracellular signaling domain is an activated intracellular domain portion, such as a T cell activation domain, which provides a primary activation signal. In some embodiments, the intracellular signaling domain includes, or additionally includes, a co-stimulatory signaling domain to promote effector function. When specifically bound to a molecule, e.g., an antigen, the receptor generally delivers an immunostimulatory signal into the cell, such as an ITAM transduction signal, thereby promoting an immune response targeting a disease or condition. In some embodiments, when the chimeric receptor is genetically engineered into immune cells, it can modulate T cell activity, and in some cases, it can modulate T cell differentiation or homeostasis, thereby resulting in genetically engineered cells with improved in vivo lifespan, viability, and / or persistence, such as for use in adoptive cell therapy methods.

[0264] Exemplary antigen receptors including CARs, and methods for manipulating and introducing such receptors into cells, are, for example, International Publication Nos. 200014257, 2013126726, 2012 / 129514, 2014031687, 2013 / 166321, 2013 / 071154, 2013 / 123061, U.S. Patent Application Publication Nos. 2002131960, 2013287748, 20130149337, U.S. Patent Publication Nos. 6, The patents described in Patent Nos. 451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353, and 8,479,118, and European Patent Application No. 2537416, and / or Sadelain This includes those described by et al., Cancer Discov. 2013 April;3(4):388-398;Davila et al.(2013)PLoS ONE 8(4):e61338;Turtle et al., Curr. Opin. Immunol., 2012 October;24(5):633-39;Wu et al., Cancer, 2012 March 18(2):160-75. In some aspects, antigen receptors include CARs described in U.S. Patent No. 7,446,190 and those described in International Publication No. 2014055668 A1.Examples of CARs include those disclosed in any of the aforementioned publications, such as International Publication No. 2014031687, U.S. Patent Nos. 8,339,645 and 7,446,179, U.S. Patent Application Publication No. 2013 / 0149337, U.S. Patent Nos. 7,446,190 and 8,389,282, Kochenderfer et al., 2013, Nature Reviews Clinical Oncology, 10, 267-276(2013); Wang et al.(2012) J. Immunother. 35(9):689-701; and Brentjens et al., Sci Transl Med. 2013 5(177). See also International Publication No. 2014031687, U.S. Patent Nos. 8,339,645 and 7,446,179, U.S. Patent Application Publication No. 2013 / 0149337, 7,446,190, and 8,389,282.

[0265] In some embodiments, engineered cells, such as T cells, express recombinant receptors, such as chimeric antigen receptors (CARs), which have specificity for specific antigens (or markers or ligands) expressed on the surface of specific cell types. In some embodiments, the antigen targeted by the receptor is a polypeptide. In some embodiments, the antigen is a carbohydrate or other molecule. In some embodiments, the antigen is selectively expressed or overexpressed on diseased or diseased cells, such as tumor or pathogenic cells, compared to normal or untargeted cells or tissues. In other embodiments, the antigen is expressed on normal cells and / or on engineered cells.

[0266] In some embodiments, the antigens targeted by the receptor include antigens associated with B-cell malignancies, such as one of several known B-cell markers. In some embodiments, the antigens targeted by the receptor are CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igκ, Igλ, CD79a, CD79b, or CD30. In certain aspects, the antigen is CD19. In some embodiments, any of such antigens is an antigen expressed on human B cells.

[0267] Chimeric receptors such as CARs generally contain an extracellular antigen-binding domain, which is one or more antigen-binding portions of an antibody molecule. In some embodiments, the antigen-binding domain is part of the antibody molecule and generally has a variable weight (V) of the antibody. H ) Chain region and / or variable light (V L ) Chain region, for example, scFv antibody fragment. In some embodiments, the antigen-binding domain is sdFv, nanobody, V H H and V NAR These are single-domain antibodies (sdAbs), such as those mentioned above. In some embodiments, the antigen-binding fragment includes an antibody variable region linked by a flexible linker.

[0268] In some embodiments, the antibody or antigen-binding fragment (e.g., scFv or V) H The domain specifically recognizes antigens such as CD19. In some embodiments, the antibody or antigen-binding fragment is derived from or a variant thereof of an antibody or antigen-binding fragment that specifically binds to CD19.

[0269] In some embodiments, the antigen is CD19. In some embodiments, the scFv is derived from an antibody or antibody fragment specific to CD19. H and V L This includes, in some embodiments, the antibody or antibody fragment that binds to CD19 is a mouse-derived antibody such as FMC63 and SJ25C1. In some embodiments, the antibody or antibody fragment is a human antibody, for example, as described in U.S. Patent Application Publication No. 2016 / 0152723.

[0270] In some embodiments, the antigen-binding domain is derived from V, FMC63. H and / or V L This includes, which in some aspects may be scFv. FMC63 generally refers to a mouse monoclonal IgG1 antibody induced against Nalm-1 and Nalm-16 cells expressing human-derived CD19 (Ling, NR, et al. (1987) Leucocyte typing III.302). In some aspects, the FMC63 antibody includes CDR-H1 and CDR-H2 shown at SEQ ID NO:38 and 39, CDR-H3 shown at SEQ ID NO:40 or 54, and CDR-L1 shown at SEQ ID NO:35, CDR-L2 shown at SEQ ID NO:36 or 55, and CDR-L3 shown at SEQ ID NO:37 or 56. The FMC63 antibody includes a heavy chain variable region (V) containing the amino acid sequence of SEQ ID NO:41. H ) and the light chain variable region (V) containing the amino acid sequence of SEQ ID NO:42 L ) includes.

[0271] In some embodiments, scFv comprises a variable light chain comprising the CDR-L1 sequence of SEQ ID NO:35, the CDR-L2 sequence of SEQ ID NO:36, and the CDR-L3 sequence of SEQ ID NO:37, and / or a variable heavy chain comprising the CDR-H1 sequence of SEQ ID NO:38, the CDR-H2 sequence of SEQ ID NO:39, and the CDR-H3 sequence of SEQ ID NO:40, or a variant of any of the sequences having 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 sequences. In some embodiments, scFv includes a variant of either the variable heavy chain region of FMC63 shown in SEQ ID NO:41 and the variable light chain region of FMC63 shown in SEQ ID NO:42, or any of the sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity. In some embodiments, the variable heavy chain and variable light chain are linked by a linker. In some embodiments, the linker is shown in SEQ ID NO:59. In some embodiments, scFv is, in order, V H , linker, and V L This includes. In some embodiments, scFv is, in order, V L , linker, and V H This includes. In some embodiments, the scFv is encoded by a nucleotide sequence shown in SEQ ID NO:57, or a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:57. In some embodiments, the scFv includes an amino acid sequence shown in SEQ ID NO:43, or a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:43.

[0272] In some embodiments, the antigen-binding domain is derived from V from SJ25C1. H and / or V L It includes, which in some aspects may be scFv. SJ25C1 is a mouse monoclonal IgG1 antibody induced against Nalm-1 and Nalm-16 cells expressing human-derived CD19 (Ling, NR, et al. (1987) Leucocyte typing III.302). In some embodiments, the SJ25C1 antibody includes the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 47-49, and the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 44-46, respectively. In some embodiments, the SJ25C1 antibody includes a heavy chain variable region (V) containing the amino acid sequence of SEQ ID NO: 50. H ) and the light chain variable region (V) containing the amino acid sequence of SEQ ID NO:51 L) comprises a variable light chain comprising the CDR-L1 sequence of SEQ ID NO:44, the CDR-L2 sequence of SEQ ID NO:45, and the CDR-L3 sequence of SEQ ID NO:46, and / or a variable heavy chain comprising the CDR-H1 sequence of SEQ ID NO:47, the CDR-H2 sequence of SEQ ID NO:48, and the CDR-H3 sequence of SEQ ID NO:49, or a variant of any of the sequences having 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 sequences. In some embodiments, scFv contains a variant of either the variable heavy chain region of SJ25C1 shown in SEQ ID NO: 50, the variable light chain region of SJ25C1 shown in SEQ ID NO: 51, or any of the sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity. In some embodiments, the variable heavy chain and variable light chain are linked by a linker. In some embodiments, the linker is shown in SEQ ID NO: 52. In some embodiments, scFv is, in order, V H , linker, and V L This includes. In some embodiments, scFv is, in order, V L , linker, and V H This includes. In some embodiments, scFv includes the amino acid sequence shown in SEQ ID NO:53, or a sequence exhibiting at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:53.

[0273] The term “antibody” as used herein is used in its broadest sense and includes intact antibodies, as well as fragment antigen-binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, and variable heavy chains (V) that can specifically bind to an antigen. H) region, single-chain antibody fragments containing single-chain variable fragments (scFv), and single-domain antibodies (e.g., sdAb, sdFv, nanobody, V) H H or V NAR This term includes polyclonal and monoclonal antibodies, including functional (antigen-binding) antibody fragments containing ) or fragments. This term also includes genetically engineered and / or otherwise modified forms of immunoglobulins, e.g., intrabodies, peptidebodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecificity, e.g., bispecific antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise specified, the term “antibody” should be understood to include its functional antibody fragment. This term also includes intact or full-length antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD. In some contexts, a CAR is a bispecific CAR, for example, containing two antigen-binding domains with different specificities.

[0274] In some embodiments, antigen-binding proteins, antibodies, and their antigen-binding fragments specifically recognize the antigen of a full-length antibody. In some embodiments, the heavy and light chains of the antibody may be full-length or may be antigen-binding portions (Fab, F(ab')2, Fv, or single-stranded Fv fragments (scFv)). In other embodiments, the constant region of the antibody heavy chain is selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, and in particular from, for example, IgG1, IgG2, IgG3, and IgG4, more specifically from IgG1 (e.g., human IgG1). In yet another embodiment, the constant region of the antibody light chain is selected from, for example, κ or λ, in particular κ.

[0275] The terms "complementarity-determining region" and "CDR," which are synonymous with "hypervariable region" or "HVR," are known to in some cases refer to non-adjacent sequences of amino acids within the antibody variable region that confer antigen specificity and / or binding affinity. Generally, each heavy chain variable region has three CDRs (CDR-H1, CDR-H2, CDR-H3), and each light chain variable region has three CDRs (CDR-L1, CDR-L2, CDR-L3). The terms "framework region" and "FR" are known to in some cases refer to the non-CDR portions of the heavy and light chain variable regions. Generally, each full-length heavy chain variable region has four FRs (FR-H1, FR-H2, FR-H3, and FR-H4), and each full-length light chain variable region has four FRs (FR-L1, FR-L2, FR-L3, and FR-L4).

[0276] The precise amino acid sequence boundaries of a given CDR or FR are as follows: Kabat et al. (1991), "Sequences of Proteins of Immunological Interest", 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); 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. This can be easily determined using one of several well-known schemes, including those described in 2003 Jan;27(1):55-77 ("IMGT" numbering scheme); Honegger A and Pluckthun 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 ("Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm", PNAS, 1989, 86(23):9268-9272 ("AbM" numbering scheme).

[0277] The boundaries of a given CDR or FR can differ 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 in both the Kabat and Chothia schemes is based on the length of the sequence of the most common antibody region, with insertions corresponding by insertion letters, e.g., "30a", and deletions appearing in some antibodies. The two schemes place specific insertions and deletions ("indels") in different positions, resulting in different numbering. The Contact scheme is based on the analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on the one used in Oxford Molecular's AbM antibody modeling software.

[0278] Table 2 below lists exemplary positional boundaries for CDR-L1, CDR-L2, CDR-L3 and CDR-H1, CDR-H2, CDR-H3 as identified by the Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering using both Kabat and Chothia numbering schemes is listed. FRs are located between CDRs; for example, FR-L1 is located before CDR-L1, FR-L2 is located between CDR-L1 and CDR-L2, and FR-L3 is located between CDR-L2 and CDR-L3, and so on. Note that because the Kabat numbering scheme shown places insertions at H35A and H35B, the ends of the Chothia CDR-H1 loop, when numbered using the Kabat numbering rules shown, will differ between H32 and H34 depending on the loop length.

[0279] (Table 2) Boundaries of CDRs under various numbering schemes TIFF0007837864000013.tif63163

[0280] Therefore, unless otherwise specified, a given antibody or its region, for example, its variable region, “CDR” or “complementarity-determining region” or individual designated CDRs (e.g., CDR-H1, CDR-H2, CDR-H3) should be understood to encompass the (or specific) complementarity-determining region as defined by one of the aforementioned schemes or other known schemes. For example, a specific CDR (e.g., CDR-H3) is a given V H or V L Where it is stated that a region contains the amino acid sequence of a corresponding CDR, such a CDR is understood to have the sequence of a corresponding CDR (e.g., CDR-H3) within a variable region, as defined by one of the aforementioned schemes or other known schemes. In some embodiments, a specific CDR sequence is specified. While the exemplary CDR sequences of the antibodies provided are represented using various numbering schemes, it is understood that the antibodies provided may contain CDRs represented according to one of the other aforementioned numbering schemes or other numbering schemes known to those skilled in the art.

[0281] Similarly, unless otherwise specified, a given antibody or its region, for example, its variable region FR or individual designated FRs (one or more) (e.g., FR-H1, FR-H2, FR-H3, FR-H4), should be understood to encompass a (or specific) framework region defined by one of the known schemes. In some cases, a scheme is specified for identifying a specific CDR(one or more) or FR(one or more), such as the Kabat, Chothia, AbM, or Contact method, or a CDR defined by another known scheme. In other cases, a specific amino acid sequence of the CDR or FR is given.

[0282] The term "variable region" or "variable domain" refers to a domain in the antibody heavy or light chain that is involved in the binding of an antibody to an antigen. (V) H and V L) generally have a similar structure, and each domain contains four conserved framework regions (FRs) and three CDRs. (See, for example, Kindt et al. Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007).) A single V H or V L The domain may be sufficient to confer antigen-binding specificity. Furthermore, the V of the antibody that binds to a specific antigen may be derived from the antibody that binds to the antigen. H or V L Isolate using the domain, and each is complementary to V L or V H You can screen domain libraries. See, for example, Portolano et al., J. Immunol. 150:880-887(1993); Clarkson et al., Nature 352:624-628(1991).

[0283] Some of the antibodies provided may contain antibody fragments. An "antibody fragment" refers to a molecule other than the intact antibody, including 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; variable heavy chains (V H ) domain, scFv and single domain V H This includes, but is not limited to, single-chain antibody molecules such as monoantibodies, as well as multispecific antibodies formed from antibody fragments. In certain embodiments, the antibody is a single-chain antibody fragment containing a variable heavy chain region and / or a variable light chain region, such as an scFv.

[0284] The term "variable region" or "variable domain" refers to a domain in the antibody heavy or light chain that is involved in the binding of an antibody to an antigen. (V) H and V L) generally have a similar structure, and each domain contains four conserved framework regions (FRs) and three CDRs. (See, for example, Kindt et al. Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007).) A single V H or V L The domain may be sufficient to confer antigen-binding specificity. Furthermore, the V of the antibody that binds to a specific antigen may be derived from the antibody that binds to the antigen. H or V L Isolate using the domain, and each is complementary to V L or V H Domain libraries can be screened. See, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0285] A single-domain antibody (sdAb) is an antibody fragment containing all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody. In some embodiments, the CAR includes an antibody heavy chain domain that specifically binds to an antigen, such as a cancer marker or targeted cell or disease, e.g., a cell surface antigen of tumor cells or cancer cells, e.g., any of the target antigens described herein or known. Exemplary single-domain antibodies include sdFv, nanobody, V H H or V NAR It includes.

[0286] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells. In some embodiments, the antibody is a recombinant fragment, such as a fragment containing a configuration that does not occur naturally, such as having two or more antibody regions or chains linked by a synthetic linker, e.g., a peptide linker, and / or a fragment that cannot be produced by enzymatic digestion of a naturally occurring intact antibody. In some embodiments, the antibody fragment is an scFv.

[0287] A “humanized” antibody is one 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 optionally contain at least a portion of the antibody constant region derived from a human antibody. A “humanized version” of a non-human antibody refers to a variant of a non-human antibody that has been humanized, typically to reduce its immunogenicity against humans while retaining the specificity and affinity of the parent non-human antibody. In some embodiments, some FR residues in a humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., an antibody from which CDR residues are derived) to restore or improve antibody specificity or affinity, for example.

[0288] In some aspects, recombinant receptors, such as chimeric antigen receptors, comprise an extracellular component containing one or more ligand (e.g., antigen) binding domains, such as antibodies or fragments thereof, and one or more intracellular signaling regions or domains (also interchangeably called cytoplasmic signaling domains or regions). In some aspects, recombinant receptors, such as CARs, further comprise a spacer and / or transmembrane domain or portion. In some aspects, the spacer and / or transmembrane domain can link the extracellular component containing the ligand (e.g., antigen) binding domain to the intracellular signaling region or domain.

[0289] In some embodiments, recombinant receptors such as CARs further include spacers, the spacers being hinge regions, e.g., the IgG4 hinge region, and / or C H 1 / C LThe immunoglobulin constant region, including and / or the Fc region, may be at least a portion of or a variant or modified version thereof. In some embodiments, the recombinant receptor further includes a spacer and / or hinge region. In some embodiments, the constant region or portion thereof is that of human IgG, such as IgG4 or IgG1. In some aspects, a portion of the constant region functions as a spacer region between the antigen-recognizing component, e.g., 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. In some examples, the spacer is 12 amino acids or about 12 amino acids long, or less than 12 amino acids long. Exemplary spacers include those having at least about 10 to 229 amino acids, at least about 10 to 200 amino acids, at least about 10 to 175 amino acids, at least about 10 to 150 amino acids, at least about 10 to 125 amino acids, at least about 10 to 100 amino acids, at least about 10 to 75 amino acids, at least about 10 to 50 amino acids, at least about 10 to 40 amino acids, at least about 10 to 30 amino acids, at least about 10 to 20 amino acids, or at least about 10 to 15 amino acids, and those containing any integer number of amino acids between either end of the enumerated range. In some embodiments, the spacer region has about 12 or fewer amino acids, about 119 or fewer amino acids, or about 229 or fewer amino acids. Exemplary spacers include an IgG4 hinge alone, an IgG4 hinge linked to the CH2 and CH3 domains, or an IgG4 hinge linked to the CH3 domain. Examples of spacers include, but are not limited to, those described in Hudecek et al. (2013) Clin. Cancer Res., 19:3153, Hudecek et al. (2015) Cancer Immunol Res. 3(2):125-135, or International Publication No. 2014031687.

[0290] In some embodiments, the spacer includes only the hinge region of IgG, such as only the hinge of IgG4 or IgG1, such as a hinge-only spacer, which is encoded by the sequence shown in SEQ ID NO:1 and SEQ ID NO:2. In some embodiments, the spacer is, for example, C H 2 and / or C H A three-domain linked Ig hinge, for example, an IgG4 hinge. In some embodiments, the spacer is C, as shown in SEQ ID NO:3. H 2 and C H A three-domain linked Ig hinge, for example, an IgG4 hinge. In some embodiments, the spacer is C, as shown in SEQ ID NO:4. H The Ig hinge is linked to only three domains, e.g., the IgG4 hinge. In some embodiments, the spacer is or includes a glycine-serine rich sequence or another flexible linker such as a known flexible linker. In some embodiments, the constant region or a portion thereof is of IgD. In some embodiments, the spacer has the sequence shown in SEQ ID NO: 5. In some embodiments, the spacer has an amino acid sequence that exhibits at least or about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with any of SEQ ID NO: 1, 3, 4, and 5.

[0291] In some aspects, the spacer may be (a) containing or consisting of all or part of an immunoglobulin hinge or a modified form thereof, or containing about 15 amino acids or fewer, and not containing the CD28 extracellular region or the CD8 extracellular region; (b) containing or consisting of an immunoglobulin hinge, optionally containing all or part of an IgG4 hinge or a modified form thereof, and / or containing about 15 amino acids or fewer, and not containing the CD28 extracellular region or the CD8 extracellular region; or (c) being 12 amino acid long or about 12 amino acid long, and / or containing or consisting of an immunoglobulin hinge, optionally containing all or part of IgG4 or a modified form thereof; or (d) SEQ ID A polypeptide spacer comprising or comprising any variant of the amino acid sequence shown in NO:1, 3-5, 27-34, or 58, or any variant of the sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the sequence, or comprising or comprising formula X1PPX2P, where X1 is glycine, cysteine, or arginine, and X2 is cysteine ​​or threonine.

[0292] In some embodiments, an antigen receptor includes an intracellular domain directly or indirectly linked to an extracellular domain. In some embodiments, a chimeric antigen receptor includes a transmembrane domain linking an extracellular domain to an intracellular signaling domain. In some embodiments, the intracellular signaling domain includes an ITAM. For example, in some aspects, the antigen recognition domain (e.g., the extracellular domain) is generally linked to one or more intracellular signaling components, e.g., in the case of CARs, a signaling component that mimics activation via an antigen receptor complex such as a TCR complex and / or signals via another cell surface receptor. In some embodiments, a chimeric receptor includes a transmembrane domain linked or fused between an extracellular domain (e.g., scFv) and an intracellular signaling domain. Thus, in some embodiments, the antigen-binding component (e.g., an antibody) is linked to one or more transmembrane domains and an intracellular signaling domain.

[0293] In one embodiment, a transmembrane domain is used that is intrinsically associated with one of the domains in a receptor, such as a CAR. In some cases, the transmembrane domain is selected or modified by amino acid substitution to minimize interaction with other members of the receptor complex by avoiding binding of such domain to transmembrane domains of the same or different surface membrane proteins.

[0294] In some embodiments, the transmembrane domain originates from either a natural or synthetic source. When the source is natural, in some aspects the domain originates from any membrane-bound or transmembrane protein. The transmembrane region includes those derived from the α, β, or ζ chains (i.e., at least their transmembrane region) of the T cell receptor, CD28, CD3ε, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137(4-1BB), or CD154. Alternatively, in some embodiments the transmembrane domain is synthetic. In some aspects the synthetic transmembrane domain primarily contains hydrophobic residues such as leucine and valine. In some aspects a triplet of phenylalanine, tryptophan, and valine is found at each end of the synthetic transmembrane domain. In some embodiments binding is via a linker, spacer, and / or transmembrane domain(s). In some aspects, the transmembrane domain includes the transmembrane portion of CD28 or a variant thereof. The extracellular domain and the transmembrane domain may be linked directly or indirectly. In some embodiments, the extracellular domain and the transmembrane domain are linked by a spacer, such as one of those described herein.

[0295] In some embodiments, the transmembrane domain of a receptor, e.g., CAR, is the transmembrane domain of human CD28 or a variant thereof, e.g., the 27-amino acid transmembrane domain of human CD28 (accession number: P10747.1), or the amino acid sequence shown in SEQ ID NO: 8, or a transmembrane domain comprising an amino acid sequence exhibiting at least or approximately 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with the sequence of SEQ ID NO: 8. In some embodiments, the transmembrane domain-containing portion of the recombinant receptor includes the amino acid sequence shown in SEQ ID NO:9, or the amino acid sequence having at least or about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with the sequence of SEQ ID NO:9.

[0296] In some embodiments, recombinant receptors, such as CARs, include at least one intracellular signaling component, such as an intracellular signaling region or domain. T cell activation is described in some aspects as being mediated by two classes of cytoplasmic signaling sequences: those that initiate antigen-dependent primary activation via the TCR (primary cytoplasmic signaling sequences), and those that act antigen-independently to provide secondary or co-stimulatory signals (secondary cytoplasmic signaling sequences). In some aspects, CARs include one or both of such signaling components. Among the intracellular signaling regions are those that mimic or approximate signals mediated by innate antigen receptors, signals mediated by such receptors in combination with co-stimulatory receptors, and / or signals mediated solely by co-stimulatory receptors. In some embodiments, there are short oligopeptide linkers or polypeptide linkers, such as those containing glycine and serine, or linkers 2 to 10 amino acid long, such as a glycine-serine doublet, that form a linkage between the transmembrane domain and the cytoplasmic signaling domain of the CAR.

[0297] In some embodiments, upon CAR ligation, the cytoplasmic domain or intracellular signaling region of the CAR activates at least one of the normal effector functions or responses of immune cells, such as T cells engineered to express the CAR. For example, in some situations, the CAR induces T cell function, such as cytolytic activity or T helper activity, such as the secretion of cytokines or other factors. In some embodiments, a truncated portion of the intracellu...

Claims

1. A pharmaceutical formulation comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19, for use in a method for treating B-cell malignancies, the method comprising: (a) the step of administering a dose of the genetically modified T cells to a subject having a B-cell malignancy; and (b) A step of administering an immunomodulatory compound to a subject, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof, step Includes, The administration of the compound to the subject is initiated within 21 days after the administration of the genetically modified T cells, and The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and A second administration period, comprising a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This is implemented in cycling regimens that include A pharmaceutical product for use in a method.

2. A pharmaceutical product comprising an immunomodulatory compound for use in a method for treating B-cell malignancies, the method being: A step comprising administering the immunomodulatory compound to a subject having a B-cell malignancy, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt thereof, solvate, hydrate, cocrystal, inclusion compound, or polymorph, Prior to the administration of the compound, the subjects were administered a dose of genetically modified T cells expressing a chimeric antigen receptor (CAR) that specifically binds to CD19. The administration of the compound to the subject is initiated within 21 days after the administration of the genetically modified T cells, and The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and A second administration period, comprising a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This is implemented in cycling regimens that include A pharmaceutical product for use in a method.

3. A pharmaceutical product comprising a dose of genetically engineered T cells expressing a CD19-specific chimeric antigen receptor (CAR) for the treatment of B-cell malignancies in a subject, used in combination with an immunomodulatory compound, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, inclusion compound, or polymorph thereof. The administration of the compound to the subject is initiated within 21 days after the administration of the genetically modified T cells, and The compound is administered daily at a dose of approximately 0.1 mg to approximately 1.0 mg / day for a maximum of 3 consecutive weeks during the first administration period. A rest period of at least one week in which the compound is not administered, beginning from the end of the first administration period, and A second administration period, comprising a 4-week cycle in which the compound is administered daily for 3 consecutive weeks at a dose of approximately 0.1 mg to approximately 1.0 mg / day in each 4-week cycle. This is implemented in cycling regimens that include A pharmaceutical product characterized by being used in such a manner.

4. The pharmaceutical product according to any one of claims 1 to 3, wherein the compound is administered in an amount of 0.3 mg to about 0.6 mg or about 0.3 mg to about 0.6 mg during the first and / or second administration period.

5. The pharmaceutical product according to any one of claims 1 to 4, wherein the first administration period begins on the same day as the administration of the genetically modified T cells.

6. The pharmaceutical product according to any one of claims 1 to 4, wherein the first administration period begins between 1 day or about 1 day and 15 days or about 15 days (including the values ​​at both ends) after administration of the genetically modified T cells.

7. The pharmaceutical product according to any one of claims 1 to 4 and 6, wherein the first administration period begins between 1 day or about 1 day and 11 days or about 11 days (including the values ​​at both ends) after administration of the genetically modified T cells.

8. The pharmaceutical product according to any one of claims 1 to 4 and 6, wherein the first administration period begins between 8 days or about 8 days and 15 days or about 15 days (including the values ​​at both ends) after administration of the genetically modified T cells.

9. The pharmaceutical product according to any one of claims 1 to 8, wherein the resting period begins on day 21 or approximately day 21 after administration of the genetically modified T cells.

10. The pharmaceutical product according to any one of claims 1 to 9, wherein a rest period is maintained until the target B cell count level recovers to the same or nearly the same level as measured before the first administration period.

11. A pharmaceutical product according to any one of claims 1 to 10, wherein the rest period is approximately one week.

12. The pharmaceutical product according to any one of claims 1 to 11, wherein the compound is administered orally during a first administration period.

13. The pharmaceutical product according to any one of claims 1 to 12, wherein the compound is a pharmaceutically acceptable salt or polymorph of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione.

14. The pharmaceutical product according to any one of claims 1 to 12, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione hydrate.

15. The pharmaceutical product according to any one of claims 1 to 12, wherein the compound is a solvate of (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione.

16. The pharmaceutical product according to any one of claims 1 to 12, wherein the compound is (S)-3-[4-(4-morpholine-4-ylmethyl-benzyloxy)-1-oxo-1,3-dihydro-isoindole-2-yl]-piperidine-2,6-dione.

17. The pharmaceutical product according to any one of claims 1 to 16, wherein the B-cell malignant tumor is lymphoma.

18. The pharmacopoeia according to claim 17, wherein the lymphoma is non-Hodgkin lymphoma (NHL), and optionally, the NHL comprises invasive NHL, diffuse large B-cell lymphoma (DLBCL), optionally transformed painless DLBCL-NOS; EBV-positive DLBCL-NOS; T-cell / histiocyte-rich large B-cell lymphoma; primary mediastinal large B-cell lymphoma (PMBCL); follicular lymphoma (FL), optionally follicular lymphoma grade 3B (FL3B); and / or high-grade B-cell lymphoma (double / triple hit) having MYC and BCL2 and / or BCL6 rearrangements with DLBCL histology.

19. A pharmaceutical product according to any one of claims 1 to 18, wherein the chimeric antigen receptor (CAR) comprises an extracellular antigen recognition domain that specifically binds to CD19 and an intracellular signaling domain that includes an immune receptor tyrosine-based activation motif (ITAM).

20. The pharmaceutical product according to claim 19, wherein the intracellular signaling domain comprises a CD3 zeta (CD3ζ) chain, optionally a signaling domain of a human CD3 zeta chain.

21. The pharmaceutical product according to claim 19 or claim 20, wherein the CAR further comprises a co-stimulatory signaling region.

22. The pharmaceutical product according to claim 21, wherein the co-stimulatory signaling region comprises a signaling domain of CD28 or 4-1BB, optionally human CD28 or human 4-1BB.

23. The pharmaceutical product according to claim 21 or 22, wherein the CAR comprises, in order, a CD19-specific single-stranded variable fragment (scFv); a transmembrane domain; a co-stimulatory signaling region; and an intracellular signaling region.

24. The pharmaceutical product according to claim 23, wherein the CAR further comprises a spacer between the transmembrane domain and the scFv.

25. The pharmaceutically acceptable agent according to claim 24, wherein the spacer comprises or consists of all or part of an immunoglobulin hinge, optionally an IgG4 hinge, or a modified form thereof, and / or comprises about 15 amino acids or less.

26. The pharmaceutical product according to claim 24 or 25, wherein the spacer comprises the amino acid sequence shown in SEQ ID NO: 1; the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 34; or a variant of any of the sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity thereto.

27. The pharmaceutical product according to any one of claims 23 to 26, wherein the transmembrane domain comprises the amino acid sequence shown in SEQ ID NO: 8 or SEQ ID NO:

9.

28. The pharmaceutical product according to any one of claims 21 to 27, wherein the co-stimulatory signaling region includes the amino acid sequence shown in SEQ ID NO: 12 or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity thereto.

29. The pharmaceutical product according to any one of claims 19 to 28, wherein the intracellular signaling region comprises an amino acid sequence shown in any one of SEQ ID NO: 13 to SEQ ID NO: 15, or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity therewith.

30. The pharmaceutical product according to any one of claims 23 to 29, wherein scFv comprises the CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), the CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and / or the CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), and / or the CDRH1 sequence of DYGVS (SEQ ID NO: 38), the CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and / or the CDRH3 sequence of YAMDYWG (SEQ ID NO: 40).

31. The pharmaceutical product according to any one of claims 23 to 30, wherein scFv comprises a heavy chain variable (VH) region containing the amino acid sequence shown as SEQ ID NO: 41, and a light chain variable (VL) region containing the amino acid sequence shown as SEQ ID NO:

42.

32. The pharmaceutical product according to any one of claims 23 to 31, wherein scFv comprises the amino acid sequence shown in SEQ ID NO:

43.

33. The dosage of the genetically engineered T cells is 1×10 8 , 7 , 8 , 7 , 8 , 7 , 8 , 7 , 6 , 8 ~5×10 8 or approximately 1×10 5 ~5×10 8 of total CAR-expressing T cells, 1×10 6 ~2.5×10 8 or approximately 1×10 6 ~2.5×10 8 of total CAR-expressing T cells, 5×10 6 ~1×10 8 or approximately 5×10 6 ~1×10 8 of total CAR-expressing T cells, 1×10 7 ~2.5×10 8 or approximately 1×10 7 ~2.5×10 8 of total CAR-expressing T cells, or 5×10 7 ~1×10 8 or approximately 5×10 7 ~1×10<00000二十>of total CAR-expressing T cells (including the values at both ends), the pharmaceutical according to any one of claims 1 to 32.

34. The pharmaceutical product according to any one of claims 1 to 33, wherein the dose of the genetically modified T cells is administered parenterally, optionally intravenously.

35. The pharmaceutical product according to any one of claims 1 to 34, wherein the genetically modified T cells are of autologous origin to the subject.

36. The pharmaceutical product according to any one of claims 1 to 34, wherein the genetically modified T cells are allogeneic to the target.

37. The pharmaceutical product according to any one of claims 1 to 36, wherein, prior to the administration of the genetically modified T cells, the subject is pre-treated with lymphocyte apheresis including the administration of fludarabine and / or cyclophosphamide.

38. Lymphocyte apheresis therapy involves approximately 200-400 mg / m². 2 Optionally, 300 mg / m² 2 Alternatively, approximately 300 mg / m² 2 Cyclophosphamide (including values ​​at both ends), and / or approximately 20–40 mg / m² 2 Optionally, 30 mg / m² 2 This may include daily administration of fludarabine for 2-4 days, or optionally 3 days, or lymphocyte apheresis at approximately 500 mg / m². 2 The pharmaceutical product according to claim 37, comprising the administration of cyclophosphamide.

39. The pharmaceutical product according to any one of claims 1 to 38, wherein the subject has received one prior treatment.

40. The pharmaceutical product according to any one of claims 1 to 38, wherein the subject has previously received more than 1, 2, 3, 4, 5, or 6 treatments.

41. The pharmacopoeia according to claim 39 or 40, wherein the prior treatment is the administration of a CD20-targeting agent and / or an anthracycline.

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