Diagnostic methods for T-cell therapy

Abstract

To provide methods of increasing the efficacy of a T cell therapy in a patient in need thereof.SOLUTION: The invention includes methods of identifying a patient who would respond well to a T cell therapy or conditioning a patient prior to a T cell therapy so that the patient responds well to a T cell therapy. The conditioning involves the steps for administering one or more preconditioning agents prior to a T cell therapy and identifying biomarker cytokines prior to administering a T cell therapy.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Statement regarding government interests This invention was developed by an agency of the U.S. Department of Health and Human Services. This was carried out in accordance with the Cooperative Research and Development Agreement with the National Cancer Institute (NCI) of the United States. The United States Government has certain rights to this invention.

[0002] Field of Invention This invention relates to a method of preconditioning a patient using one or more preconditioning agents that have the ability to increase the serum levels of one or more cytokines that exhibit efficacy in T-cell therapy. This relates to a method for identifying patients suitable for T-cell therapy through conditioning, or a method for preparing patients to be suitable for T-cell therapy.

[0003] Background of the Invention Human cancers, by their very nature, are composed of normal cells that have undergone genetic or epigenetic transformation to become abnormal cancer cells. In this process, cancer cells begin to express proteins and other antigens different from those expressed by normal cells. The body's innate immune system can utilize these abnormal tumor antigens to specifically target and kill cancer cells. However, cancer cells employ various mechanisms to prevent immune cells, such as T lymphocytes and B lymphocytes, from effectively targeting and killing them.

[0004] Human T-cell therapy involves concentrating cells to target and kill cancer cells in patients. Alternatively, it relies on modified human T cells. This is to enrich the concentration of native T cells that have the ability to target tumor antigens, or to genetically modify T cells to specifically target known cancer antigens. Various techniques have been developed to target tumors. These therapies have been found to have a mild but promising effect on tumor size and patient survival. However, it has been found difficult to predict whether a given T-cell therapy will be effective in each individual patient. Therefore, it is necessary to identify patients who will respond well to T-cell therapy, or to prepare patients to respond well to T-cell therapy.

[0005] Summary of the Invention This disclosure is, (i) Interleukin-15 ("IL-15"), interleukin-7 ("IL-7"), and monocytes Chemotactic protein 1 ("MCP-1"), C-reactive protein ("CRP"), placental growth factor ("PLGF"), interferon-gamma-inducible protein 10 ("IP-10"), and any combination thereof. Increase the serum level of at least one additional cytokine selected from the group consisting of the following: The stage of administering one or more preliminary conditioning agents to the patient that have the ability to do so, Bini (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. The present invention provides a method for treating cancer in patients suitable for T-cell therapy, including [specific example].

[0006] This disclosure increases serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. The step involves administering to the patient one or more pre-conditioning agents that have the ability to enhance the condition. A method for treating cancer in patients suitable for T-cell therapy, including IL-15, IL-7 and when the serum levels of at least one additional cytokine are elevated, This further provides ways in which patients are treated with T-cell therapy.

[0007] This disclosure is, (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Increase the serum levels of at least one additional cytokine selected from the following groups. The step of administering one or more preliminary conditioning agents to the patient that have the ability to do so, (ii) The stage of administering an additional amount of one or more preconditioning agents, This involves administering an effective dose of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. Furthermore (iii) The patient has elevated blood levels of IL-15, IL-7, and at least one additional cytokine. If the levels are clear, this is the stage to administer T-cell therapy. The invention also provides methods for treating cancer in patients suitable for T-cell therapy, including those mentioned above.

[0008] This disclosure increases serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. The step involves administering to the patient one or more pre-conditioning agents that have the ability to enhance the condition. This also includes providing methods for identifying patients suitable for T-cell therapy.

[0009] This disclosure is, (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Increase the serum levels of at least one additional cytokine selected from the following groups. The step of administering one or more preliminary conditioning agents to the patient that have the ability to do so, and to (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. The report also provides a method for identifying patients suitable for T-cell therapy, including [specific example].

[0010] This disclosure is, (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Increase the serum levels of at least one additional cytokine selected from the following groups. The step of administering one or more preliminary conditioning agents to the patient that have the ability to do so, (ii) The stage of administering an additional amount of one or more preconditioning agents, This step involves administering an effective dose of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. Rabini (iii) The patient has elevated blood levels of IL-15, IL-7, and at least one additional cytokine. If the levels are clear, this is the stage to administer T-cell therapy. The report also provides a method for identifying patients suitable for T-cell therapy, including [specific example].

[0011] This disclosure is, The stage of administering one or more preliminary conditioning agents to the patient. To precondition patients requiring T-cell therapy, a selection is made from a group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. A method for increasing serum levels of at least one additional cytokine. This also provides a method of treatment with T-cell therapy when a patient exhibits elevated serum levels of IL-15, IL-7, and at least one additional cytokine.

[0012] This disclosure relates to IL-15, used for preconditioning patients requiring T-cell therapy. From the group consisting of IL-7, MCP-1, CRP, PLGF, IP-10, and any combination thereof A method for increasing serum levels of at least one additional selected cytokine. And, (i) The step of administering one or more preconditioning agents to the patient, (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. We also provide methods that include this.

[0013] This disclosure relates to a group of T-cells consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof, for preconditioning patients requiring T-cell therapy. A method for increasing serum levels of at least one additional selected cytokine. And, (i) The step of administering one or more preliminary conditioning agents to the patient, (ii) The stage of administering an additional amount of one or more preconditioning agents, This step involves administering an effective dose of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. Rabini (iii) The patient has elevated blood levels of IL-15, IL-7, and at least one additional cytokine. If the levels are clear, this is the stage to administer T-cell therapy. We also provide methods that include this.

[0014] In certain embodiments, the method disclosed herein involves one or more preconditioning The procedure further includes measuring serum levels of IL-15, IL-7, and at least one cytokine after administration of the drug.

[0015] In a particular embodiment, one or more of the preconditioning agents disclosed herein are used. The material comprises clophosphamide and a purine analog. In one embodiment, the purine analog is selected from pentostatin and fludarabine.

[0016] In certain aspects, T-cell therapy comprises a group consisting of tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, modified autologous cell therapy (eACT), allogeneic T-cell transplantation, and any combination thereof. Selected. [Brief explanation of the drawing]

[0017] [Figure 1] An example of a CAR-modified T cell and a schematic diagram of its structure are shown. In this exemplary CAR-modified T cell, the target-binding domain contains an antibody-derived scFv domain, the costimulatory domain is derived from CD28, and the essential activation domain is derived from CD3ζ (zeta). The viral vector can hold the CAR vector construct, which can then be incorporated into the T cell genome. The T cell can then express the CAR construct as a transmembrane protein. [Figure 2A] This figure shows the disease response of patients after treatment with anti-CD-19 CAR+ T cells. Figure 2A shows the best response in patients with B-cell malignancy as a percentage change in disease status. Shaded bars indicate complete response (CR). Shaded bars indicate partial response. White bars indicate stable disease (SD). Black bars indicate progressive disease (PD). [Figure 2B]This figure shows the disease response of patients after treatment with anti-CD-19 CAR+ T cells. Figure 2B shows the patient's disease response as a number of months after CAR+ T cell infusion. Black bars indicate partial response (PR), and gray bars indicate complete response (CR). A break in a bar marked "PD" indicates that the patient experienced progressive disease. Inverted triangles mark the time of T cell infusion. Black circles indicate the time of B cell recovery. White circles indicate the time of CAR+ T cell elimination from the patient's blood. Horizontal arrows indicate that the patient's response is ongoing. [Figure 3] This provides a sample diagram of a Phase 1 clinical trial aimed at determining the safety, efficacy, and dose-limiting toxicity of treating patients with 500 mg / m2 / day of cyclophosphamide, 30 mg / m2 / day of fludarabine, and 2 × 10⁶ cells / kg of anti-CD19 CAR+ T cells. [Figure 4] The serum levels of selected cytokine analytes before and after conditioning with 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine are shown. Serum levels of interleukin-15 (IL-15; Figure 4A), monocyte chemotactic protein 1 (MCP-1; Figure 4B), gamma-inducible protein 10 (IP-10; Figure 4C), placental growth factor (PLGF; Figure 4D), soluble intercellular adhesion molecule 1 (sICAM-1; Figure 4E), C-reactive protein (CRP; Figure 4F), vascular endothelial growth factor D (VEGF-D; Figure 4G), and macrophage inflammatory protein 1β (MIP-1b; Figure 4H) are shown before and after administration of 300 mg / m2 of cyclophosphamide and 30 mg / m2 of fludarabine. Pre-administration serum was collected on days -12 to -5, and post-administration serum was collected on day 0, before the administration of T-cell therapy (Figures 4A-4H). [Figure 5]Figure 5A shows the ratio of changes in serum levels of selected cytokine analytes after conditioning with 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine in patients who responded to or did not respond to subsequent T-cell therapy. Figure 5H shows the ratio of changes in serum levels of IL-15 (Figure 5A), MCP-1 (Figure 5B), IP-10 (Figure 5C), PLGF (Figure 5D), sICAM-1 (Figure 5E), CRP (Figure 5F), VEGF (Figure 5G), and MIP-1b (Figure 5H) for both responders and non-responders. Horizontal lines represent the mean (Figures 5A-H). Figure 5A shows the IL-15 changes for individual patients, with each patient's disease responsiveness indicated next to each data point as partial response (PR), complete response (CR), stable disease (SD), or progressive disease (PD) (Figure 5A). [Figure 6-1] The serum concentrations of selected cytokine analytes, measured at various time points from day -10 to day 18, are shown for patients who received 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine before receiving T-cell therapy on day 0. Serum concentrations of granulocyte-macrophage colony-stimulating factor (GM-CSF; Figure 6A), IL-2 (Figure 6B), MCP-1 (Figure 6C), IL-6 (Figure 6D), IL-10 (Figure 6E), MCP-4 (Figure 6F), CRP (Figure 6G), interferon-gamma (IFNγ; Figure 6H), granzyme A (Figure 6I), IL-15 (Figure 6J), IL-5 (Figure 6K), and granzyme B (Figure 6L) are shown. [Figure 6-2] The serum concentrations of selected cytokine analytes, measured at various time points from day -10 to day 18, are shown for patients who received 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine before receiving T-cell therapy on day 0. Serum concentrations of IL-8 (Figure 6M), IP-10 (Figure 6N), MIP-1b (Figure 6O), and PLGF (Figure 6P) are shown. [Figure 6-3]The serum concentrations of selected cytokine analytes, measured at various time points from day -10 to day 18, are shown for patients who received 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine prior to T-cell therapy on day 0. Serum concentrations of IL-16 (Figure 6Q), thymic and activating regulatory chemokines (TARC; Figure 6R), eotaxin-3 (Figure 6S), and sICAM-1 (Figure 6T) are shown. [Figure 6-4] The serum concentrations of selected cytokine analytes, measured at various time points from day -10 to day 18, are shown for patients who received 300 mg / m2 / day of cyclophosphamide and 30 mg / m2 / day of fludarabine prior to T-cell therapy on day 0. Serum concentrations of soluble vascular adhesion molecule 1 (sVCAM; Figure 6U) and (SAA; Figure 6V) are also shown. [Figure 7] Serum concentrations of selected cytokine analytes, measured before and after administration of cyclophosphamide at 300 mg / m2 / day and fludarabine at 30 mg / m2 / day, are shown. Post-administration serum was collected immediately before T-cell infusion. Serum concentrations of IL-15 (Figure 7A), IL-7 (Figure 7B), PLGF (Figure 7C), CRP (Figure 7D), IL-5 (Figure 7E), IL-10 (Figure 7F), MCP-1 (Figure 7G), IP-10 (Figure 7H), and sICAM-1 (Figure 7I) are shown. Each data point represents a single patient. Horizontal bars indicate the mean (Figures 7A-7I). Wilcoxon's matched-pair signed-rank test p-values ​​were applied to the analytes measured before and after conditioning, and the corresponding p-values ​​are shown (Figures 7A-7I). Some IL-7 values ​​exceeded the upper limit of quantification (ULOQ; Figure 7B). [Figure 8]This report shows the in vitro production of various cytokine analytes by anti-CD19 CAR+ T cells (K562-CD19) after stimulation with K562 cells, compared to a negative control (K562-NGFR). The concentrations of GM-CSF (Figure 8A), IL-2 (Figure 8B), IFNγ (Figure 8C), IL-5 (Figure 8D), IL-4 (Figure 8E), IL-13 (Figure 8F), tumor necrosis factor alpha (TNFα; Figure 8G), IL-6 (Figure 8H), granzyme B (Figure 8I), MIP-1β (Figure 8J), MIP-1α (Figure 8K), and soluble CD137 (Figure 8L) are shown for both control T cells and anti-CD19 CAR+ T cells. T1, T2, and immunoconstitutive cytokines (Figures 8A-8F), as well as pro-inflammatory cytokines and chemokines (Figures 8G-8L), are appropriately labeled. Product T cells were co-incubated with K562-CD19 cells or control K562-NGFR cells, and data were collected before injection by measuring the concentrations of the described analytes in the culture medium (Figures 8A-8L). [Figure 9] The percentage of anti-CD19 CAR+ T cells (K562-CD19) expressing various cytokines after association with the target antigen is shown compared to the negative control (K562-NGFR). The percentage of cells expressing CD107α (Figure 9A), 4-1BB (Figure 9B), and programmed death 1 (PD-1; Figure 9C) is also shown. Pre-injection data were collected by co-incubating product T cells with K562-CD19 cells or control K562-NGFR cells and measuring the concentrations of selected activation markers in the culture medium (Figures 9A-9C). The p-values ​​shown represent the results of paired t-tests comparing K562-CD19 test cells with K562-NGFR negative control cells (Figures 9A-9C). [Figure 10]This report describes various characteristics of product T cells and peripheral blood lymphocytes (PBLs) in terms of production time (days). Data include: the percentage of anti-CD-19 CAR+ T cells detected in the product compared to PBLs; the ratio of CD8 to CD4 in the product compared to PBLs; the relative occurrences of naive T cells, central memory (Tcm) T cells, effector memory (Tem) T cells, and effector (Teff) T cells within the anti-CD19 CAR+ CD8+ T cell population; and the relative occurrences of naive T cells, central memory (Tcm) T cells, effector memory (Tem) T cells, and effector (Teff) T cells within the anti-CD19 CAR+ CD4+ T cell population (Figure 10). Phenotypic analysis of product T cells before injection and PBLs at peak proliferation in the blood was performed for anti-CD19 CAR+ T cells (Figure 10). The p-value represents the result of a rank test of association between production time and the composition of the T cell subset. [Figure 11] The following shows the expression profiles of cytokines, chemokines, and other markers observed after conditioning in NHL patients according to the present invention. CRP: C-reactive protein. PLGF: Placental growth factor. MCP-1: Monocyte chemotactic protein-1. [Figure 12] This document quantifies the changes observed in cytokines, chemokines, and other markers after conditioning using cyclophosphamide and fludarabine according to the present invention. [Figure 13] This shows the magnitude of changes in circulating IL-15 and perforin after conditioning chemotherapy associated with objective response. P values ​​were not adjusted for multiplicity. Analysis was performed for markers measured before CAR T cell infusion. [Figure 14]This report presents biomarker analyses of cytokines, chemokines, and effector molecules. Within each biomarker category, markers were ranked from lowest to highest p-value using the Wilcoxon signed-rank test. Biomarkers that were altered in most patients and had a p-value <0.05 are shown. Of the 41 markers measured, only 7 showed alteration in most patients with a p-value <0.05. Analysis was performed for markers measured before CAR T-cell infusion. [Figure 15] This study demonstrates the sequential induction and elimination of immunohomeostatic, inflammatory, and modulating cytokines, chemokines, and immune effector molecules. Representative markers are shown. Of the 41 markers measured, 22 in total—IL-15, IL-7, IL-2, granzyme B, granzyme A, CRP, IL-6, GM-CSF, IL-5, IFNg, IL-10, MCP-1, MCP-4, IP-10, IL-8, TARC, MIP1a, MIP1b, PLGF, VEGF-D, sICAM-1, and FGF-2—showed elevations of at least twofold above baseline levels in at least 50% of patients after CAR T-cell treatment. Peak formation was observed for immunohomeostatic cytokines and chemokines on days 3–4. [Figure 16] This study demonstrates the sequential induction and elimination of immunomodulatory, inflammatory, and regulatory cytokines, chemokines, and immune effector molecules. Representative markers are shown. Of the 41 markers measured, 22 in total—IL-15, IL-7, IL-2, granzyme B, granzyme A, CRP, IL-6, GM-CSF, IL-5, IFNg, IL-10, MCP-1, MCP-4, IP-10, IL-8, TARC, MIP1a, MIP1b, PLGF, VEGF-D, sICAM-1, and FGF-2—showed elevations at least twice the baseline level after CAR T-cell treatment in at least 50% of patients. Peak formation was observed for immunomodulatory cytokines and chemokines on days 5–7. "ULOQ": Upper limit of quantification. [Figure 17]This report shows the changes in treatment-related biomarkers and clinical responses induced by anti-CD19 CAR T cells according to the present invention. The maximum change in marker levels after CAR T cell treatment compared to baseline (pre-conditioning) is shown. Each row represents an individual subject. The maximum change in marker levels was compared between the responder and non-responder groups for all 41 biomarkers evaluated using the Wilcoxon rank-sum test. Only biomarkers with p<0.10 are shown, without adjusting for multiplicity. The p-values ​​for IL-7 and sICAM-1 were <0.05. This association was also applicable to changes at the absolute level of IL-7 (p=0.0165), IL-15 (p=0.0314), and IL-15 (p=0.041). [Figure 18-1] The changes in analyte levels before and after conditioning with cyclophosphamide and fludarabine are shown. Figures 18A–18F show the pre- and post-levels of IL-15 (Figure 18A), IP-10 (Figure 18B), CRP (Figure 18C), IL-7 (Figure 18D), MCP-1 (Figure 18E), and perforin (Figure 18F). [Figure 18-2] The changes in analytes before and after conditioning with cyclophosphamide and fludarabine are shown. Figure 18G summarizes the changes in serum levels for various analytes and their corresponding p-values. [Figure 19] For IL-15 (Figure 19A), IP-10 (Figure 19B), and perforin (Figure 19C), the correlation between changes at the analyte level after conditioning and objective response to CAR T-cell therapy is shown. Figure 19D provides a summary of the statistical significance of the data provided for each of Figures 19A-19C. [Modes for carrying out the invention]

[0018] Detailed description of the invention The present invention relates to a method for identifying patients suitable for T-cell therapy, such as modified CART cell therapy, such as autologous cell therapy (eACT®), and then treating the patients with T-cell therapy. Thus, this method has the ability to increase the serum levels of certain cytokines, such as IL-15, IL-7, and at least one additional biomarker cytokine. The procedure may include a step of preconditioning the patient by administering multiple preconditioning agents. Preconditioning the patient before T-cell therapy with one or more preconditioning agents improves the efficacy of T-cell therapy by reducing the number of endogenous lymphocytes and increasing serum levels of homeostatic cytokines and / or immunostimulants present in the patient, including IL-15 and IL-7. The implanted T cells proliferate after being administered to the patient, creating a more optimal microenvironment.

[0019] The present invention (i) The patient receives certain cytokines, e.g., IL-15, IL-7, and at least one One or more that have the ability to increase serum levels of additional biomarker cytokines The stage of administering multiple pre-conditioning drugs; (ii) The step of administering to the patient one or more cytokines, e.g., IL-15, IL-7, and at least one additional biomarker cytokine, which have been shown to be associated with increased efficacy of T-cell therapy; or (iii) The patient receives additional treatment, such as additional cytokines, additional doses of one or more preconditioning agents, or one or more cytokines. A step in which T cells are administered, wherein the additional treatment increases serum levels of one or more cytokines, e.g., IL-15, IL-7, and at least one additional biomarker cytokine, which have been identified as being associated with the increased efficacy of T cell therapy. stage Methods for creating an environment more suitable for T-cell therapy in patients requiring such therapy, including , further concerning.

[0020] definition To make this disclosure easier to understand, certain terms are defined first. Each of the following terms used herein shall have the meanings set forth below, unless otherwise expressly provided herein. Additional definitions are set forth throughout this application.

[0021] As used herein, the term "and / or" should be understood to specifically disclose each of two particular features or components together with or without the other. Therefore, phrases such as "A and / or B" in this specification should be understood as... The term "and / or" as used is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, "A, B, and / or C" The term "and / or" as used in such phrases encompasses each of the following situations: It is intended to be: A, B, and C; A, B, or C; A or C; A or B; B or C; A and and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0022] Whenever a situation is described using the word “contains” in this specification, other similar situations described using the terms “consist of” and / or “essentially become from” are also provided. It is understood that this is the case.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as they would ordinarily be understood by a person skilled in the art relating to this disclosure. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised, 2000, Oxford University Press are intended to be used by a person skilled in the art relating to this disclosure. It provides a general dictionary of many words.

[0024] Units, prefixes, and symbols are presented in the form recognized by the International System of Units (SI). Numerical ranges include the number defining that range. The headings provided herein are not limitations on the various aspects of this disclosure that can be seen by reference to this specification as a whole. Thus, the terms defined immediately below are more fully defined by reference to the entire specification.

[0025] The term "activation" refers to the state of immune cells, such as T cells, that have been sufficiently stimulated to induce detectable cell proliferation. Activation is characterized by induced cytokine production and detectable It can also be related to the effects function. The term "activated T cell" is, among other things, a cell segment. This represents a T cell that has undergone cleavage.

[0026] "Administer" refers to the introduction of a drug into the body of a subject using any of the various methods and delivery systems known to those skilled in the art. Exemplary routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other extra-intestinal routes, e.g., by injection or infusion. As used herein, "extra-intestinal administration" means a method of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroporation. In some embodiments, the formulations are administered via non-extra-intestinal routes, e.g., orally. Other non-extra-intestinal routes include topical, epidermal, or mucosal routes, e.g., intranasal, intravaginal, rectal, sublingual, or topical. The administration can be, for example, a single dose, multiple doses, and / or one or more long-term doses. .

[0027] As used herein, “adverse event” (AE) refers to any undesirable, generally unintended or undesirable sign (including abnormal laboratory findings), symptom, medical occurrence, or use of a medical procedure. It is a disease related to the use of the medication. The definition of an adverse event includes exacerbation of a pre-existing medical condition. Exacerbation is an increase in the severity, frequency, and / or duration of a pre-existing medical condition, or a worsening of the condition. This indicates that it is related to the outcome.

[0028] The term “antibody” (Ab) non-limitingly includes immunoglobulins, which are glycoproteins that specifically bind to an antigen. Generally, an antibody may contain at least two heavy (H) chains and two light (L) chains or their antigen-binding moieties, linked together by disulfide bonds. Each H chain includes a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three constant domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated as VL herein) and a light chain constant region. The light chain constant region comprises one The constant domain, including the CL. The VH and VL regions are superimposed on the complementarity-determining region (CDR). The variable regions can be further subdivided into larger, conserved regions called framework regions (FRs) scattered between them. Each VH and VL contains three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of Ab may mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1q).

[0029] Immunoglobulins may be derived from any of the generally known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. Subclasses of IgG are also well known to those skilled in the art, and they include This includes, non-limitingly, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to a class or subclass of Ab (e.g., IgM or IgG1) encoded by a heavy chain constant region gene. The term "antibody" includes, by example, both natural and non-natural Abs; monoclonal and polyclonal Abs; chimeric and humanized Abs; human or non-human Abs; fully synthetic Abs; and single-chain Abs. Non-human Abs may be humanized by recombinant methods, which may reduce their immunogenicity in humans. Unless explicitly stated, and unless the context indicates otherwise, the term "antibody" also includes any antigen-binding fragment or portion of any of the aforementioned immunoglobulins, including monovalent and bivalent fragments or portions, as well as single-chain Abs.

[0030] An "antigen-binding molecule" or "antibody fragment" represents any portion of an antibody that is not the whole. Antigen-binding molecules may include antigen complementation-determining regions (CDRs). Examples include, non-limitingly, Fab, Fab', F(ab')2, and Fv fragments, dAb, and linear antibodies. This includes scFv antibodies and multispecific antibodies formed from antigen-binding molecules.

[0031] An "antigen" refers to any molecule that has the ability to trigger an immune response or to be bound by an antibody. An immune response may involve either antibody production or activation of specific immune-qualified cells, or both. Those skilled in the art will readily understand that virtually any macromolecule, including proteins or peptides, can act as an antigen. Antigens can be endogenously expressed, i.e., expressed by genomic DNA, or they can be recombinantly expressed. Antigens can be found in cancer cells, for example. They may be specific to certain tissues or may be widely expressed. In addition, relatively large molecular fragments can act as antigens. In one embodiment, the antigen is a tumor antigen.

[0032] The term "self" refers to any material originating from the same individual that is later reintroduced into that individual. For example, the manipulated autologous cell therapy (eACT®) described herein involves collecting lymphocytes from a patient, manipulating those lymphocytes to express, for example, a CAR construct, and then reintroducing them to the same patient. This includes being administered and then returned.

[0033] The term "same species" refers to any material originating from one individual that is then introduced into another individual of the same species. This represents the material used, and an example is allogeneic T cell transplantation.

[0034] "Cancer" refers to a broad group of diseases characterized by the uncontrolled proliferation of abnormal cells in the body. Unregulated cell division and proliferation can lead to the formation of malignant tumors that invade adjacent tissues and may metastasize to distant parts of the body through the lymphatic system or bloodstream. "Cancer" or "cancer tissue" may include tumors. Examples of cancers that can be treated by the methods of the present invention include, but are not limited to, lymphoma, leukemia, and cancers of the immune system, including other leukemias and malignancies. In some embodiments, the methods of the present invention can be used to treat, for example, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma (NHL), mediastinal large B-cell lymphoma (PMBC), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), transformed follicular lymphoma, and splenic marginal lamina. Lymphoma (SMZL), esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non-T cell ALL), chronic lymphocytic leukemia (CLL), pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma Cancers, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, and asbestos-induced cancers Tumors originating from environment-induced cancers, other B-cell malignancies, and combinations of said cancers. It can be used to reduce tumor size. Certain cancers may be responsive to chemotherapy or radiotherapy, or the cancer may be anti-therapeutic. Anti-therapeutic cancers represent cancers that are not correctable by surgical intervention, and the cancer is either unresponsive to chemotherapy or radiotherapy from the beginning, or becomes unresponsive over time.

[0035] As used herein, “antitumor effect” refers to a biological effect that may manifest as a reduction in tumor volume, a reduction in the number of tumor cells, a reduction in tumor cell proliferation, a reduction in the number of metastases, an increase in overall survival or progression-free survival, an increase in life expectancy, or an improvement in various physiological symptoms associated with tumors. Antitumor effect may also refer to the prevention of tumor development, such as a vaccine.

[0036] In this specification, the term "progression-free survival," which may be abbreviated as PFS, is used in relation to treatment. From the date of establishment to the date of disease progression or death from any cause according to the revised IWG Response Criteria for Malignant Lymphoma. This indicates the time until the date.

[0037] "Disease progression" should be determined by measurement of malignant lesions using radiographic imaging or other methods and should not be reported as an adverse event. Death due to disease progression in the absence of signs and symptoms should be reported as the type of primary tumor (e.g., DLBCL).

[0038] As used herein, "DOR," which can be abbreviated as DOR, refers to the first Based on objective response, confirmed by the revised IWG criteria for evaluating treatment efficacy for malignant lymphoma. This represents the period of time until the date of disease progression or death.

[0039] The term "overall survival," which can be abbreviated as OS, is defined as the time from the date of treatment to the date of death.

[0040] As used herein, "cytokine" refers to a single cytokine that reacts to contact with a specific antigen. This refers to non-antibody proteins released by cells, in which case cytokines are second cells It interacts with and mediates the response in the second cell. Cytokines are intrinsically transmitted by the cell. Cytokines can be expressed sexually or administered to the target. Cytokines are macrophages, B cells, T cells. Cytokines can be released by cells and immune cells, including mast cells, to propagate immune responses. Cytokines can induce a variety of responses in recipient cells. Cytokines can include homeostatic cytokines, chemokines, pro-inflammatory cytokines, effectors, and acute-phase proteins. For example, homeostatic cytokines, including interleukin (IL) 7 and IL-15, can promote the survival and proliferation of immune cells, while pro-inflammatory cytokines can promote inflammatory responses. Examples of homeostatic cytokines include, but are not limited to, IL-2, IL-4, IL-5, IL-7, IL-10, IL-12p40, IL-12p70, IL-15, and interferon (IFN) gamma. Examples of pro-inflammatory cytokines include, but are not limited to, IL-1a, IL-1b, IL-6, IL-13, IL-17a, tumor necrosis factor (TNF)-alpha, TNF-beta, fibroblast growth factor (FGF)2, granulocyte-macrophage colony-stimulating factor (GM-CSF), soluble intercellular adhesion molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D, and placental growth factor (PLGF). Examples of effectors include, but are not limited to, granzyme A. It contains granzyme B, soluble Fas ligand (sFasL), and perforin. Examples of proteins include, non-limitingly, C-reactive protein (CRP) and serum amyloid A (SAA).

[0041] "Chemokines" are a type of cytokine that mediates chemotaxis or directional movement of cells. Examples of chemokines include, non-limitingly, IL-8, IL-16, eotaxin, eotaxin-3, This includes macrophage-derived chemokines (MDC or CCL22), monocyte chemotactic protein 1 (MCP-1 or CCL2), MCP-4, macrophage inflammatory protein 1α (MIP-1α, MIP-1a), MIP-1β (MIP-1b), gamma-inducible protein 10 (IP-10), and thymic and activation-regulating chemokines (TARC or CCL17).

[0042] Other examples of analytes and cytokines of the present invention include, but are not limited to, chemokine (CC motif) ligands (CCL) 1, CCL5, monocyte-specific chemokine 3 (MCP3 or CCL7), monocyte chemotactic protein 2 (MCP-2 or CCL8), CCL13, IL-1, IL-3, IL-9, IL-11, IL-12, IL-14, IL-17, IL-20, IL-21, granulocyte colony-stimulating factor (G-CSF), leukemia suppressor (LIF), oncostatin M (OSM), CD154, lymphotoxin (LT) beta, 4-1BB ligand (4-1BBL), proliferation-inducing ligand (APRIL), CD70, CD153, CD178, glucocorticoid-inducing TNFR-related ligand (GITRL), tumor Necrosis factor superfamily member 14 (TNFSF14), OX40L, TNF and ApoL-associated leukocyte This includes either the current ligand 1 (TALL-1) or the TNF-associated apoptosis-inducing ligand (TRAIL).

[0043] As used herein, the terms “serum level” and “serum concentration” are interchangeable and refer to the amount of analyte in the serum of interest. The serum level of a given analyte can be measured using any method known in the art. For example, the serum level of cytokines can be measured using enzyme-linked immunosorbent assay (ELISA). In a particular embodiment... The serum levels of cytokines can be measured using the EMDmillipore LUMINEX® xMAP® multiplex assay.

[0044] As used herein, “dosing interval” means the amount of time that elapses between the administration of multiple doses of the formulation disclosed herein to a subject. Therefore, the dosing interval may be expressed as a range.

[0045] The dosages described herein are either "weight-based dosage" or "body surface area (BSA)-based dosage." It can be expressed as follows: Weight-based dose is the dose administered to the patient, calculated based on the patient's weight, e.g., mg / kg. BSA-based dose is the dose administered to the patient, calculated based on the patient's surface area, e.g., mg / m². 2 Regarding administration to humans, by weight The two forms of dose measurement can be converted by multiplying the based dose by 37 or dividing the BSA-based dose by 37. For example, a dose of cyclophosphamide of 60 mg / kg administered to a human subject is equivalent to a dose of 2220 mg / m² of the same drug administered to the same subject. 2 It corresponds to.

[0046] As used herein, the term "number of doses" refers to the number of times a dose of the formulation disclosed herein is administered within a given time period. The number of doses may be expressed as the number of doses per unit of time. For example, a pre-conditioning agent, such as cyclophosphamide, may be administered once daily for each of five consecutive days, once daily for each of four consecutive days, and so on for three consecutive days. It can be administered once a day, once a day for two consecutive days, or once a day. Furthermore, a second pre-conditioning agent, such as fludarabine, is administered once a day for each of the following 8 consecutive days, once a day for each of the following 7 consecutive days, once a day for each of the following 6 consecutive days, once a day for each of the following 5 consecutive days, once a day for each of the following 4 consecutive days, and for each of the following 3 consecutive days. Each can be administered once a day, once a day for two consecutive days, or once a day. Yes, it is possible. In other embodiments, fludarabine is administered once daily for five consecutive days or once daily for three consecutive days.

[0047] The “therapeutic effective dose,” “effective dose,” “effective amount,” or “therapeutic effective dosage” of a drug, preconditioning agent, or therapeutic agent, such as engineered CAR T cells, is any amount of the drug, used alone or in combination with another therapeutic agent, that protects a subject from disease development or promotes disease regression, as demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or the prevention of functional impairment or disability caused by the suffering of the disease. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to experienced practitioners, for example, in human subjects during clinical trials, in animal model systems to predict efficacy in humans, or by assaying the activity of the drug in in vitro assays.

[0048] As used herein, the term "lymphocyte" includes natural killer (NK) cells, T cells, and other related terms. This includes cells, or B cells. NK cells are a key component of the intrinsic immune system. NK cells are a type of harmful (cytotoxic) lymphocyte. They reject tumor and virus-infected cells. They work through apoptosis or programmed cell death processes. NK cells are named "natural killers" because they do not require activation to kill cells. T cells play a major role in cellular immunity (without the involvement of antibodies). Their T cell receptors (TCRs) distinguish self from other lymphocyte types. The thymus, a specialized organ of the immune system, is primarily responsible for... They are responsible for the maturation of T cells. There are six types of T cells, namely helper T cells (e.g., CD4+ cells), cytotoxic T cells (TC, cytotoxic T lymphocyte, CTL, T-killer cell, cell-lysing T cell, also known as CD8+ T cell or killer T cell), and memory T cells ((i) memory T cells like naive cells). SCM Stem cells include CD45RO-, CCR7+, CD45RA+, CD62L+ (L-selectin), CD27+, CD28+, and These cells are IL-7Rα+, but they also produce large amounts of CD95, IL-2Rβ, CXCR3, and LFA-1. (ii) Central memory T CM The cells express L-selectin and CCR7, and these cells secrete IL-2 but not IFNγ or IL-4, (iii) however, Effector Memory T EM Cells include regulatory T cells (Treg, suppressor T cells, or CD4+CD25+ regulatory T cells), natural killer T cells (NKT), and gamma delta T cells (which do not express L-selectin or CCR7, but rather produce effector cytokines such as IFNγ and IL-4). On the other hand, B cells are involved in humoral immunity (antibodies are involved). In mammals, it plays a major role. It produces antibodies and antigens, acts as an antigen-presenting cell (APC), and transforms into a memory B cell after being activated by antigen interaction. Immature B cells are formed in the bone marrow, from which the name originates.

[0049] The term “genetic manipulation” or “manipulation” refers, non-limitingly, to methods of altering the genome of a cell, including deleting coding regions or non-coding regions or parts thereof, or inserting coding regions or parts thereof. In some embodiments, the cells being modified are lymphocytes, such as T cells, which can be obtained from either a patient or a donor. For example Chimeric antigen receptors (CARs) or T cell receptors (TCRs) are incorporated into the cell's genome. Cells can be modified to express exogenous constructs that can be introduced.

[0050] "Immune response" refers to the immune system's cells (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, and neutrophils) and soluble macromolecules (abs, cytokines, etc.) produced by any of these cells or by the liver. The action of the complement system, which removes invasive pathogens and cells infected with pathogens from the body of vertebrates. Alternatively, in the case of tissue, cancer cells or other abnormal cells, or in the case of autoimmune or pathological inflammation, it exhibits selective targeting, binding to, damage to, destruction of, and / or removal of normal human cells or tissues.

[0051] The term “immunotherapy” refers to the treatment of subjects suffering from a disease or at risk of developing or relapsing the disease, by a method that includes steps of inducing, enhancing, suppressing, or otherwise modifying the immune response. Examples of immunotherapy include, but are not limited to, T-cell therapy. T-cell therapy may include adoptive T-cell therapy, tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, modified autologous cell therapy (eACT), and allogeneic T-cell transplantation. However, as those skilled in the art will know, The detailed disclosure acknowledges that the conditioning method enhances the effectiveness of any transplanted T-cell therapy. Examples of T-cell therapy are described in U.S. Patent Publication Nos. 2014 / 0154228 and 2002 / 0006409, U.S. Patent No. 5,728,388, and International Publication No. 2008 / 081035. Yes, they are.

[0052] T cells for immunotherapy can be derived from any known origin in this technology. Yes, T cells can be differentiated in vitro from hematopoietic stem cell populations, or T cells can be obtained from subjects such as peripheral blood mononuclear cells, bone marrow, and lymphocytes. T cells can be obtained from nodule tissue, umbilical cord blood, thymic tissue, tissue from infection sites, ascites, pleural fluid, splenic tissue, and tumors. In addition, T cells can be derived from one or more T cell lines available in the art. T cells are collected from blood units of a subject using any number of techniques known to a skilled technician, e.g., FICOLL® isolation and / or apheresis. They can also be obtained from. An additional method for isolating T cells for T cell therapy is disclosed in U.S. Patent Publication No. 2013 / 0287748, which is incorporated herein by reference in its entirety.

[0053] The term "manipulated autologous cell therapy," also known as adoptive cell transplantation and abbreviated as "eACT(trademark)," involves the collection of the patient's own T cells, which are then used to recognize and target one or more antigens expressed on the surface of one or more specific tumor cells or malignant disease cells. This is a process of genetic modification. T cells can be manipulated to express, for example, a chimeric antigen receptor (CAR) or a T cell receptor (TCR). CAR-positive (+) T cells are The CAR is engineered to express an extracellular single-chain variable fragment (scFv) specific to a particular tumor antigen, which is linked to an intracellular signaling region containing a co-stimulatory domain and an activation domain. The co-stimulatory domain may be derived from, for example, CD28, and the activation domain may be derived from, for example, CD3-zeta (Figure 1). In certain embodiments, the CAR may have two, three, four, or It is designed to have a co-stimulatory domain beyond that. CAR scFv is designed for use in all normal B cells, and not limited to B cells, including B cells with malignancies such as NHL, CLL, and non-T cell ALL. It is possible to design the CAR+ T-cell therapy to target CD19, a transmembrane protein expressed by cells of a specific cell lineage. Examples of CAR+ T-cell therapies and constructs are described in U.S. Patent Publications 2013 / 0287748, 2014 / 0227237, 2014 / 0099309, and 2014 / 0050708, which are incorporated by reference in their entirety.

[0054] As used herein, the term “patient” includes any human being suffering from cancer (e.g., lymphoma or leukemia). The terms “subject” and “patient” are used interchangeably herein.

[0055] The terms "peptide," "polypeptide," and "protein" are used interchangeably to refer to compounds composed of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and There is no limit to the maximum number of amino acids that can make up a polypeptide or peptide sequence. A polypeptide contains two or more amino acids linked to each other by peptide bonds. This includes any peptide or protein. As used herein, the term refers to both short chains, also commonly called peptides, oligopeptides, and oligomers in the art, and longer chains, of which numerous types exist, commonly referred to as proteins in the art. "Polypeptides" include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, heterologous polypeptides, modified polypeptides, derivatives, analogs, and fusion proteins. Polypeptides include native peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0056] As used herein, "stimulus" refers to the primary response induced by the binding of a stimulating molecule to its cognitive ligand, in which case the binding mediates a signaling event. "Stimulating molecule" refers to a molecule on a T cell, for example, a cognitive stimulating ligand present on an antigen-presenting cell. It is a T cell receptor (TCR) / CD3 complex that specifically binds to the ligand. A "stimulating ligand" is a ligand that, when present on antigen-presenting cells (e.g., aAPCs, dendritic cells, B cells, etc.), specifically binds to stimulating molecules on T cells, thereby mediating a primary response by T cells, including, but not limited to, activation, initiation of an immune response, and proliferation. This includes, non-limitingly, peptide-loaded MHC class I molecules, anti-CD3 antibodies, and superagonists. This includes anti-CD28 antibodies and super-agonist anti-CD2 antibodies.

[0057] As used herein, "co-stimulatory signals" refer to signals such as TCR / CD3 ligation. Combined with the primary signal, it non-limitingly promotes proliferation and / or upregulates key molecules. It represents signals that lead to T cell responses such as ration or downregulation. .

[0058] The term "co-stimulatory ligand" as used herein refers to cognitive costimulatory components on T cells. It contains molecules on antigen-presenting cells that specifically bind to the cell. The binding of a co-stimulatory ligand provides signals that mediate T cell responses, including proliferation, activation, and differentiation, non-limitingly. A stimulating ligand induces a signal, which is then transmitted by the stimulating molecule, for example, to T cells. In addition to the primary signal provided by the binding of the receptor (TCR) / CD3 complex to peptide-loaded major histocompatibility complex (MHC) molecules, co-stimulatory ligands include, non-limitingly, CD7, B7-1 (CD80), B7-2 (CD86), programmed death (PD)L1, PD-L2, 4-1BB ligand, and OX40 ligand. Gand, inducible costimulatory ligand (ICOS-L), intercellular adhesion molecule (ICAM), CD30 ligand, CD40, CD70, CD83, human leukocyte antigen G (HLA-G), MHC class I chain-related protein A (MICA), MHC class I chain-related protein B (MICB), herpesvirus entry vector (HVEM), lymphotoxin base Receptor, 3 / TR6, immunoglobulin-like transcript (ILT) 3, ILT4, Toll ligand receptor It includes an agonist or antibody that binds to B7-H3, and a ligand that specifically binds to B7-H3. This is possible. Co-stimulatory ligands can be non-limited, including co-stimulatory molecules present on T cells, for example, non Specifically, antibodies that specifically bind to CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, tumor necrosis factor superfamily member 14 (TNFSF14 or LIGHT), natural killer cell receptor C (NKG2C), B7-H3, and CD83. It contains ligands that specifically bind to it.

[0059] "Co-stimulatory molecules" specifically bind to costimulatory ligands, thereby enabling T cells to co-stimulate each other. Stimulus response, for example, non-limitingly, mediating proliferation, through cognitive binding partners on T cells Co-stimulatory molecules include, but are not limited to, CD27, CD28, 4-1BB, OX40, CD30, CD40, CD83, PD-1, ICOS, LFA-1, CD2, CD7, TNFSF14(LIGHT), NKG2C, B7-H3, MHC class 1 molecules, B and T lymphocyte attenuators (BTLA), and Toll ligand receptors.

[0060] The terms “conditioning” and “pre-conditioning” are used interchangeably herein and refer to preparing a patient to an appropriate state for T-cell therapy. Conditioning as used herein includes, but is not limited to, prior to T-cell therapy. To reduce the number of endogenous lymphocytes, to eliminate cytokine sink, or one of the following This involves increasing serum levels of multiple biomarker cytokines or pro-inflammatory factors, enhancing the effector function of T cells administered after conditioning, and antigen Enhancing the activation and / or utilization of presented cells, or any combination thereof. This includes. In one embodiment, "conditioning" means one or more cytokines, For example, this includes increasing serum levels of interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-10 (IL-10), interleukin-5 (IL-5), gamma-inducing protein 10 (IP-10), interleukin-8 (IL-8), monocyte chemotactic protein 1 (MCP-1), placental growth factor (PLGF), C-reactive protein (CRP), soluble intercellular adhesion molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), or any combination thereof. In another embodiment, “conditioning” includes increasing serum levels of IL-7, IL-15, IP-10, MCP-1, PLGF, CRP, or any combination thereof.

[0061] The terms “reduce” and “decrease” are used interchangeably herein and refer to any change that is less than the initial value. “Reduce” and “decrease” are relative terms requiring a comparison between before and after measurement. “Reduce” and “decrease” include total depletion.

[0062] "Treatment" or "to treat" a subject refers to any type of intervention or process performed on the subject or administration of an active agent to the subject for the purpose of reducing, alleviating, restoring, inhibiting, slowing or preventing the onset, progression, development, severity or recurrence of symptoms, complications or conditions, or disease-related biochemical features. In one embodiment, "treatment" or "to treat" includes a partial response. In another embodiment, "treatment" or "to treat" includes a complete response.

[0063] The use of options (e.g., "or") indicates one of the options, both, or any combination thereof. It should be understood to mean either "1" or "1". The indefinite article "1" used in this specification "a" or "an" means "one or one of any of the listed or enumerated components." It should be understood as representing "multiple" or "plural".

[0064] The terms “approximately” or “essentially composed of” refer to a value or composition that is within an acceptable margin of error for a particular value or composition as determined by those skilled in the art, and which in part depends on how that value or composition is measured or determined, i.e., the limits of the measuring system. For example, “approximately” or “essentially composed of” may mean within or exceeding one standard deviation for implementations in the art. Alternatively, “approximately” or “essentially composed of” may mean a range of up to 10% (i.e., ±10%). For example, approximately 3 mg may include any number between 2.7 mg and 3.3 mg (about 10%). It is possible. Furthermore, especially with respect to biological systems or biological processes, these terms can mean up to one order of magnitude or up to five times the value. Where a particular value or composition is provided in the application and claims, unless otherwise stated, the meaning of “about” or “essentially composed of” should be assumed to be within an acceptable margin of error for that particular value or composition.

[0065] Any concentration range, percentage range, ratio range, or integer range described herein is, Any integer within the specified range, and, where appropriate, its fraction unless otherwise specified (for example) Please understand that this includes values ​​that are one-tenth and one-hundredth of an integer.

[0066] Various aspects of the present invention are described in further detail in the following subsections.

[0067] Method of invention This invention identifies patients suitable for T-cell therapy after an initial preliminary conditioning phase. The present invention is directed toward a method of stratifying patients into subgroups after the initial preliminary conditioning stage and treating the subgroups in the appropriate next stage. One group of patients identified by this method is the first group of patients after the preliminary conditioning stage. After administering the conditioning method, some patients may be suitable for T-cell therapy without any additional preconditioning. Another group of patients undergoes initial preconditioning. This group may be unsuitable for staged T-cell therapy and may require a second preliminary conditioning stage. Patients in the third group may be unsuitable for T-cell therapy even after the subsequent preliminary conditioning stage. The present invention uses one or more preliminary conditioning stages. This also includes preparing patients for T-cell therapy by increasing certain biomarker cytokines in them.

[0068] This invention confirms that increased expression of certain cytokines in patients who have received a pre-conditioning regimen indicates increased efficacy of T-cell therapy. The cytokines that demonstrate increased efficacy in T-cell therapy include IL-15, IL-7, and monocyte chemotactic protein 1 ("MCP-1"), C-reactive protein ("CRP"), placental growth factor ("PLGF"), interferon-gamma-inducible protein 10 ("IP-10"), and any combination thereof. Includes at least one additional cytokine that is further selected. In one embodiment, biomer In another embodiment, the biomarker cytokines are IL-15, IL-7, and MCP-1. In yet another embodiment, the biomarker cytokines are IL-15, IL-7, and CRP. In yet another embodiment, the biomarker cytokines are IL-15, IL-7, and PLGF. In yet another embodiment, the biomarker Sexual cytokines are IL-15, IL-7, MCP-1, and IP-10. In other embodiments, Iomarker cytokines are IL-15, IL-7, MCP-1, and CRP. In some cases, the biomarker cytokines are IL-15, IL-7, MCP-1, and PLGF. In certain embodiments, the biomarker cytokines are IL-15, IL-7, IP-10, and CRP. In other embodiments, the biomarker cytokines are IL-15, IL-7, PLGF, and IP-10. In yet another embodiment, the biomarker cytokines are IL-15, IL-7, MCP-1, IP-10, and CRP. In yet another embodiment, the biomarker cytokines are IL-15, IL-7, MCP-1, IP-10, and PLGF. In a particular embodiment, the biomarker cytokines The cytokines are IL-15, IL-7, MCP-1, CRP, and PLGF. In some embodiments, Biomarker cytokines include IL-15, IL-7, IP-10, CRP, and PLGF. In this embodiment, the biomarker cytokines are IL-15, IL-7, IP-10, MCP-1, CRP, and PLGF.

[0069] In addition to increased serum expression of biomarker cytokines, patients suitable for T-cell therapy The additional features are (i) a reduced number of endogenous lymphocytes; (ii) an increased number of T cells. (iii) Effect function; (iii) Increased activation and / or availability of antigen-presenting cells; or (iv) Any combination thereof may also be shown.

[0070] Endogenous lymphocytes, reduced by the pre-conditioning method, can non-limitingly inhibit the antitumor effect of adoptive transplanted T cells, endogenous regulatory T cells, B cells, and natural lymphocytes. This may include RAL killer cells, CD4+ T cells, CD8+ T cells, or any combination thereof. Endogenous lymphocytes may compete with adoptive-transplanted T cells for sensitization to antigens and supportive cytokines. One or more preconditioning agents are used. Pretreatment clears this competition and raises the levels of endogenous cytokines, including IL-15 and IL-7. This leads to an increase. After adoptive T cells are administered to a patient, they increase in level. Exposed to endogenous IL-15 and IL-7, as well as other homeostatic cytokines or pro-inflammatory factors Furthermore, preconditioning treatment can lead to tumor cell death and an increase in tumor antigens in the patient's serum. Without being bound by any theory, non-limitingly, one or more preconditioning treatments including cyclophosphamide and purine analogs may be used. Conditioning with conditioning agents promotes homeostatic proliferation, activation, and The immune environment is modified through the induction of IL-15, IL-7, and one or more other biomarker cytokines that may be favorably transported.

[0071] In one embodiment, the present invention provides at least one additional biomarker selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. One or more preliminary conditions that have the ability to increase cytokine serum levels The method includes a step of administering a suppository to the patient, and a method for identifying patients suitable for T-cell therapy. nothing.

[0072] In another embodiment, the present invention is (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Serum of at least one additional biomarker cytokine selected from the following groups Administer the patient one or more pre-conditioning agents that have the ability to increase the level. The stage of doing so, (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. This includes a method for identifying patients suitable for T-cell therapy.

[0073] The present invention (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Serum of at least one additional biomarker cytokine selected from the following groups Administer the patient one or more pre-conditioning agents that have the ability to increase the level. At the stage of doing so, (ii) A step of measuring serum levels of IL-15, IL-7, and at least one additional biomarker cytokine, and (iii) The patient has IL-15, IL-7, and at least one additional biomarker cytoka The stage to administer T-cell therapy when elevated serum levels of iodine are observed. This also includes methods for identifying patients suitable for T-cell therapy.

[0074] In certain embodiments, the present invention is (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Increase at least one additional biomarker cytokine selected from the following groups. The step of administering one or more preliminary conditioning agents to the patient that have the ability to induce this condition, (ii) The stage of administering an additional amount of one or more preconditioning agents, This includes effective amounts of IL-15, IL-7, and / or MCP-1, CRP, PLGF, IP-10, and their At least one additional biomarker selected from a group consisting of any combination The stage of directly administering tokines, and (iii) The patient has IL-15, IL-7, and at least one additional biomarker cytoka The stage to administer T-cell therapy when elevated serum levels of iodine are observed. This also includes methods for identifying patients suitable for T-cell therapy.

[0075] In some embodiments, the present invention is (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Serum of at least one additional biomarker cytokine selected from the following groups Administer the patient one or more pre-conditioning agents that have the ability to increase the level. At the stage of doing so, (ii) A step of measuring serum levels of IL-15, IL-7, and at least one additional biomarker cytokine, (iii) The step of administering an additional amount of one or more preconditioning agents, or an effective amount of IL-15, IL-7, and / or MCP-1, CRP, PLGF, IP-10, and their At least one additional biomarker selected from a group consisting of any combination The stage of directly administering tokines, (iv) Optionally measuring serum levels of IL-15, IL-7, and at least one additional biomarker cytokine, and (v) The patient has IL-15, IL-7, and at least one additional biomarker-mediated cytometry The stage to administer T-cell therapy when elevated serum levels are observed. This also includes methods for identifying patients suitable for T-cell therapy.

[0076] The present invention The stage of administering one or more preliminary conditioning agents to the patient. To precondition patients requiring T-cell therapy, a selection is made from a group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. Increase the serum levels of at least one additional biomarker cytokine. A method for treating a patient with T-cell therapy when the patient exhibits elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. It also includes.

[0077] In one embodiment, the present invention preconditions patients who require T-cell therapy. Therefore, serum samples containing at least one additional biomarker cytokine selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. A method for increasing the level, (i) The step of administering one or more preconditioning agents to the patient, (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. Includes methods.

[0078] In another aspect, the present invention preconditions patients who require T-cell therapy. Therefore, blood samples containing at least one additional biomarker cytokine selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. A method for increasing the purity level, (i) The step of administering one or more preliminary conditioning agents to the patient, (ii) The stage of administering an additional amount of one or more preconditioning agents, This involves an effective amount of IL-15, IL-7, and at least one additional biomarker cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. The stage of administration, and (iii) The patient has IL-15, IL-7, and at least one additional biomarker cytoka The stage to administer T-cell therapy when elevated serum levels of iodine are observed. This includes methods that include [specific methods].

[0079] In a particular embodiment, this method involves administering one or more preconditioning agents. The procedure then further includes measuring serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. In one embodiment, serum levels of IL-15 are measured. In another embodiment, serum levels of IL-7 are measured. In another embodiment, serum levels of IL-15 and IL-7 are measured. The level is measured.

[0080] This invention relates to patients who exhibit elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine after administration of one or more preconditioning agents. The present invention also provides a method for treating a person, which includes the step of administering T-cell therapy to the patient. The method includes a step of further preconditioning patients who do not show sufficient serum levels of IL-15, IL-7, and at least one additional biomarker cytokine after administration of a first dose of one or more preconditioning agents, the step of administering a second dose of one or more preconditioning agents. After administration of the conditioning agent, any IL-15, IL-7, and at least additional bacteria In certain patients who do not exhibit elevated serum levels of biomarker cytokines, the present invention further includes the step of directly administering an effective amount of biomarker cytokines to the patient.

[0081] In other embodiments, the present invention relates to a patient after administration of one or more preconditioning agents. The process includes identifying one or more cytokines that are elevated in the serum, where the elevated serum levels of one or more cytokines indicate an increase in subsequent T-cell therapy. It correlates with the responsiveness. In one embodiment, serum levels of one or more cytokines are predictable. It can be measured after administration of a conditioning agent. One or more cytokines Patients who do not show elevated serum levels of IL-15 may be subjected to additional treatment. In one embodiment, the additional treatment may involve the administration of certain cytokines, such as IL-15, IL-7, and less IL-15. At the very least, it has the ability to increase the serum levels of one additional biomarker cytokine. The procedure includes administering to the patient one or more preliminary conditioning agents in a second dose. In another embodiment, the additional treatment involves administering one or more additional cytokines to the patient previously One or more treatments to increase serum levels of one or more cytokines, including the step of administering to the patient one or more preconditioning agents that have not been administered, or one or more cytokines, such as IL-15, IL-7, and T cells engineered to express at least one additional biomarker cytokine, to the patient. This includes the administration stage.

[0082] In another aspect, the present invention upregulates in patients before the administration of T-cell therapy. If present, methods for identifying cytokines that increase the effectiveness of T-cell therapy. And, with or without administering other pre-conditioning agents to the patient, T cells Prior to therapy, the patient is administered the identified cytokine, or one or more pre-conditioning agents are used to further increase the serum levels of the identified cytokine. The present invention provides a method for increasing the quantity. In another embodiment, the present invention provides a method for continuously monitoring the serum level of a particular biomarker cytokine during conditioning and adjusting the timing and dosage accordingly, for example, preliminary conditioning until a desired serum level of the biomarker cytokine is achieved and the patient is ready for T-cell therapy. It provides a way to keep going.

[0083] In other embodiments, the present invention includes a method for increasing serum levels of one or more biomarker cytokines in patients requiring T-cell therapy. In some embodiments, 1 Serum levels of one or more biomarker cytokines indicate that the patient has one or more It increases by administering a pre-conditioning agent. In some embodiments, one Alternatively, serum levels of multiple biomarker cytokines are increased by administering one or more cytokines to the patient, selected from IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. In some embodiments, one or more biomarker cytokines Serum levels of itkaine include IL-15, IL-7, as well as MCP-1, CRP, PLGF, IP-10, and so on. At least one additional cytokine selected from the group consisting of any combination of these This is increased by administering T cells engineered to express one or more selected cytokines to the patient.

[0084] In one embodiment, the present invention provides one or more preconditioning cells capable of increasing serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. A method for treating cancer in a patient suitable for T-cell therapy, comprising the step of preconditioning the patient by administering a conditioning agent to the patient, wherein the patient exhibits elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. The present invention includes a method of treatment with T-cell therapy in some cases. The present invention has the ability to increase serum levels of one or more additional biomarker cytokines selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. The step of preconditioning the patient using multiple preconditioning agents enhances the effectiveness of T-cell therapy subsequently administered to the patient. In another embodiment, the present invention This increases serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. A method for treating cancer in a patient suitable for T-cell therapy, comprising the step of administering to the patient one or more capable preconditioning agents, wherein the patient has IL-15, IL-7, and increased serum levels of at least one additional biomarker cytokine When indicated, this includes methods treated with T-cell therapy.

[0085] In other embodiments, the present invention is (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Serum of at least one additional biomarker cytokine selected from the following groups Administer the patient one or more pre-conditioning agents that have the ability to increase the level. The stage of doing so, (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional biomarker cytokine. The invention includes a method for treating cancer in patients suitable for T-cell therapy, including [specific example].

[0086] In another embodiment, the present invention is (i) IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof Serum of at least one additional biomarker cytokine selected from the following groups Administer the patient one or more pre-conditioning agents that have the ability to increase the level. At the stage of doing so, (ii) The stage of administering an additional amount of one or more preconditioning agents, This involves an effective amount of IL-15, IL-7, and at least one additional biomarker cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. The stage of administration, and (iii) The patient has IL-15, IL-7, and at least one additional biomarker cytoka The stage to administer T-cell therapy when elevated serum levels of iodine are observed. The invention includes a method for treating cancer in patients suitable for T-cell therapy, including [specific example].

[0087] In another embodiment, the present invention is (i) Give the patient one or more pre-conditioning agents (e.g., cyclophosphamide) The stage of administering (and / or fludarabine), (ii)IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1 The steps include measuring serum levels of perforin, MIP-1b, or any combination thereof, and (iii) One or more preconditioning agents (e.g., cyclophosphamide and / After administration of (or fludarabine), the subject has elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, e.g., IL-15, IP-10, and / or IL-7, as well as / or perforin and / Alternatively, if a reduced serum level of MIP-1b is observed, one or more preliminary conditions The stage of characterizing the ning agent as effective in preparing targets for T-cell therapy. The method includes identifying a dose of one or more preconditioning agents effective for preparing a subject for T-cell therapy, including IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or those. Any combination, for example, elevated serum levels of IL-15, IP-10, and / or IL-7, Patients showing reduced serum levels of rabine and / or perforin and / or MIP-1b The method further comprises administering one or more preconditioning agents. In other embodiments, the method further comprises administering T-cell therapy to a patient exhibiting elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, e.g., IL-15, IP-10, and / or IL-7, and / or decreased serum levels of perforin and / or MIP-1b.

[0088] In another embodiment, the present invention is (i) Give the patient one or more pre-conditioning agents (e.g., cyclophosphamide) The stage of administering (and / or fludarabine), (ii)IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1 The steps include measuring serum levels of perforin, MIP-1b, or any combination thereof, and (iii) After administration of one or more preconditioning agents, the subject is IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any of the above. The step of characterizing one or more preconditioning agents (e.g., cyclophosphamide and / or fludarabine) as effective for preparing a subject for T-cell therapy when a combination, for example, shows elevated serum levels of IL-15, IP-10, and / or IL-7, and / or decreased serum levels of perforin and / or MIP-1b. The method includes verifying the efficacy of one or more preconditioning agents to prepare a subject for T-cell therapy, including IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any of the same. If the combination of the following is true, for example, elevated serum levels of IL-15, IP-10, and / or IL-7, then In patients who have decreased serum levels of perforin and / or MIP-1b, 1 The method further comprises administering one or more preconditioning agents. In other embodiments, the method further comprises administering T-cell therapy to a patient who exhibits elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, e.g., IL-15, IP-10, and / or IL-7, and / or decreased serum levels of perforin and / or MIP-1b.

[0089] In some embodiments, the administration of one or more preconditioning agents is performed on the patient. The number of endogenous lymphocytes is reduced. In certain embodiments, endogenous lymphocytes include regulatory T cells, B cells, natural killer cells, CD4+ T cells, CD8+ T cells, or any combination thereof. In some embodiments, one or more preconditioning agents are administered. This increases the utilization of homeostatic cytokines, such as IL-15 and / or IL-7. In some embodiments, administration of one or more preconditioning agents improves the condition The effector function of T cells administered after conditioning is enhanced. In some embodiments, the administration of one or more preconditioning agents activates and / or utilizes antigen-presenting cells. Strengthen your abilities.

[0090] In certain embodiments, the administration of one or more preconditioning agents is one or Compared to the antitumor efficacy of T-cell therapy without the administration of multiple pre-conditioning agents, improvement was observed. It induces the antitumor efficacy of T-cell therapy.

[0091] Cytokine levels This invention relates to biomarker-based cytokine for effective cancer treatment in T-cell therapy. The use of the in is described. In particular, this application provides a suitable environment for transplanted T cells. Identify the cytokine group, which is a key factor in this process, and improve the efficacy of T-cell therapy. In aspects, the present invention relates to inducing the upregulation of biomarker cytokines or increasing their serum levels. Administration of one or more preconditioning agents prior to T-cell therapy increases the levels of biomarker cytokines and modifies the immune environment in a manner favorable to the homeostatic proliferation, activation, and transport of T cells. After the transplanted T cells were administered to the patient, they showed increased levels of endogenous cytotoxicity. They are exposed to the cytokine. Biomarker cytokines that indicate the efficacy of T cells are not limited to This includes at least one additional biomarker cytokine selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof. .

[0092] This invention relates to the use of biomarker cytokines in patients requiring T-cell therapy. This also includes methods for increasing the function. In a particular embodiment, the biomarker cytokine is IL-15, IL-7, and at least one additional biomarker cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof.

[0093] In certain embodiments, the present invention is (i) Biomarkers after administration of one or more preliminary conditioning agents in a first dose The steps include measuring serum levels of sex cytokines (e.g., IL-15 or IL-7), and (ii) The stage of administering T-cell therapy to patients who show elevated levels of one or more biomarker cytokines (e.g., patients who are more likely to respond to T-cell therapy). The present invention provides a method for determining treatment options for a patient, including the following: (i) Biomarkers after administration of one or more preliminary conditioning agents in a first dose The steps include measuring serum levels of sex cytokines (e.g., IL-15 or IL-7), and (ii) Patients who do not show increased levels of one or more biomarker cytokines Patients who exhibit one or more biomarker cytokines below threshold levels. One or more spare doses of the second dose for patients (e.g., patients less likely to respond to T-cell therapy) The stage of administering conditioning agents. The present invention provides a method for determining treatment options for a patient, including the following: (i) A step of measuring serum levels of biomarker cytokines (e.g., IL-15 or IL-7) after administration of one or more preconditioning agents, (ii) Patients who do not show increased levels of one or more biomarker cytokines Patients who exhibit one or more biomarker cytokines below threshold levels. The step of administering one or more biomarker cytokines, (iii) The stage of administering T-cell therapy to patients who show increased levels of one or more biomarker cytokines or who show one or more cytokines above threshold levels. This provides a method for determining treatment options for a patient, including [specific examples of treatment options].

[0094] In one embodiment, the serum IL-15 level after administration of one or more preconditioning agents is increased by more than 5 times, more than 10 times, more than 15 times, more than 20 times, or more than 25 times. An increase exceeding approximately 30 times, 35 times, 40 times, 45 times, 50 times, 60 times, 70 times, 80 times, or 90 times. This indicates that patients are more likely to respond to T-cell therapy. In certain embodiments, serum IL-15 levels increase by more than approximately 10 times after administration of one or more preconditioning agents. The increase indicates that the patient is more likely to respond to T-cell therapy. In another aspect, the serum IL-15 levels increased approximately 20 times after administration of one or more preconditioning agents. An increase exceeding this indicates that the patient is more likely to respond to T-cell therapy. In another aspect, A more than 30-fold increase in serum IL-15 levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. To increase serum IL-15 levels, one or more doses of exogenous IL-15 and / or additional amounts may be used. A preliminary conditioning agent can be administered to the patient.

[0095] In another embodiment, the level of serum IL-7 after administration of one or more preconditioning agents An increase of more than approximately 2 times, more than 3 times, more than 4 times, more than 5 times, more than 10 times, more than 15 times, more than 20 times, more than 25 times, more than 30 times, more than 35 times, more than 40 times, more than 45 times, more than 50 times, more than 60 times, more than 70 times, more than 80 times, or more than 90 times indicates that the patient is likely to respond to T-cell therapy. It indicates a higher quality. In certain embodiments, one or more preconditioning agents An increase of more than twofold in serum IL-7 levels after administration indicates that the patient is more likely to respond to T-cell therapy. To increase serum levels of IL-7, exogenous IL-7 and / Alternatively, administer one or more additional doses of a pre-conditioning agent to the patient. It is possible.

[0096] In other embodiments, serum IP-10 levels after administration of one or more preconditioning agents exceed approximately 2 times, approximately 3 times, approximately 4 times, approximately 5 times, or approximately 6 times. more than 7 times, more than 8 times, more than 9 times, more than 10 times, more than 15 times An increase of more than approximately 20 times, or more than approximately 30 times, indicates that the patient is likely to respond to T-cell therapy. It indicates a higher level. In certain embodiments, the administration of one or more preconditioning agents A more than twofold increase in serum IP-10 levels after administration suggests that the patient may be responding to T-cell therapy. This indicates a higher quality. In another embodiment, one or more preconditioning agents A more than threefold increase in serum IP-10 levels after administration indicates that the patient is responding to T-cell therapy. It indicates a higher probability. In another embodiment, one or more preconditioning agents An increase of more than four times in serum IP-10 levels after administration indicates that the patient is responding to T-cell therapy. This indicates a higher probability. In another embodiment, one or more preconditioning An increase of more than 7 times in serum IP-10 levels after administration of the drug indicates that the patient is responding to T-cell therapy. This indicates a higher probability. To increase serum levels of IP-10, the patient may be administered exogenous IP-10 and / or additional amounts of one or more preconditioning agents.

[0097] In some embodiments, serum MCP-1 levels after administration of one or more preconditioning agents are increased by more than 1.5 times, more than 2 times, more than 3 times, more than 4 times, or more than 5 times. An increase of more than double, more than approximately 6 times, more than approximately 7 times, more than approximately 8 times, more than approximately 9 times, more than approximately 10 times, more than approximately 15 times, or more than approximately 20 times indicates that the patient may be responding to T-cell therapy. It indicates a higher probability. In other embodiments, one or more preconditioning agents A more than twofold increase in serum MCP-1 levels after administration indicates that the patient is responding to T-cell therapy. This indicates a higher probability. In another embodiment, one or more preconditioning A more than threefold increase in serum MCP-1 levels after administration of the drug indicates that the patient is responding to T-cell therapy. This indicates a higher probability of [something happening]. In another embodiment, one or more preconditioning A more than fivefold increase in serum MCP-1 levels after administration of the drug indicates that the patient is responding to T-cell therapy. This indicates a higher probability of [something happening]. In another embodiment, one or more preconditioners An increase of more than 7 times in serum MCP-1 levels after administration of the suppository indicates that the patient is responding to T-cell therapy. This indicates a higher probability of answering. Exogenous MCP to increase serum levels of MCP-1 -1 and / or additional amounts of one or more pre-conditioning agents may be administered to the patient.

[0098] In a particular embodiment, serum PLGF after administration of one or more preconditioning agents At the level, more than 1.5 times, more than 2 times, more than 3 times, more than 4 times, more than 5 More than double, more than approximately 10 times, more than approximately 15 times, more than approximately 20 times, more than approximately 25 times, more than approximately 30 times, more than approximately 35 times, more than approximately 40 times, more than approximately 45 times, more than approximately 50 times, more than approximately 60 times, more than approximately 70 times, more than approximately 80 times, more than approximately 90 times, or more than approximately 100 times The increase indicates that the patient is more likely to respond to T-cell therapy. In certain embodiments, serum PLGF levels increased by approximately 1.5 times after administration of one or more preconditioning agents. An increase exceeding this indicates that the patient is more likely to respond to T-cell therapy. In another aspect, A more than twofold increase in serum PLGF levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. In another aspect, A more than threefold increase in serum PLGF levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. To increase the bell, the patient may be administered exogenous PLGF and / or additional amounts of one or more preconditioning agents.

[0099] In other embodiments, serum CRP levels after administration of one or more preconditioning agents exceed approximately 1.5 times, approximately 2 times, approximately 3 times, approximately 4 times, and approximately 5 times. It is more than 9 times, more than 10 times, more than 15 times, more than 20 times, more than 25 times If it is more than 30 times, more than 35 times, more than 40 times, more than 45 times, more than 50 times, more than 60 times, more than 70 times, more than 80 times, more than 90 times, or more than 100 times A more than doubling increase indicates that patients are more likely to respond to T-cell therapy. In one embodiment, an increase of more than approximately 1.5 times in serum CRP levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. In another embodiment, an increase of more than approximately 2 times in serum CRP levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. In yet another embodiment, an increase of more than approximately 5 times in serum CRP levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. A more than doubling increase indicates that patients are more likely to respond to T-cell therapy. Another aspect Then, approximately 9 in serum CRP levels after administration of one or more preconditioning agents. A more than doubling increase indicates that patients are more likely to respond to T-cell therapy. Another aspect Therefore, an increase of more than approximately 10 times in serum CRP levels after administration of one or more preconditioning agents indicates that the patient is more likely to respond to T-cell therapy. Another aspect So, in patients suitable for T-cell therapy, the CRP level is one or more of a pre-condition. Serum CRP levels increased by at least approximately 25 times after administration of the chemotherapeutic agent. This indicates that the patient is more likely to respond to T-cell therapy. To increase serum CRP levels, exogenous CRP and / or an additional dose of one or more pre-conditioning A stimulant can be administered to the patient.

[0100] In some embodiments, one or more preconditioning agents are interleuk Interleukin-10 ("IL-10"), Interleukin-5 ("IL-5"), Interleukin-8 ("IL-8"), Soluble Further increases serum levels of sex cell adhesion molecule 1 ("sICAM-1"), soluble vascular adhesion molecule 1 ("sVCAM-1"), or any combination thereof.

[0101] In some embodiments, serum levels of any one or more cytokines are measured on one or more days selected from the day of administration of one or more preconditioning agents and from the day of administration of T-cell therapy to 21 days after the administration of T-cell therapy.

[0102] Those skilled in the art will, without limitation, use one or more of the preconditioning agents described herein. Use, administration of one or more exogenous cytokines to a patient as described herein, one or more to induce the expression of one or more endogenous cytokines or to prevent their degradation It is recognized that levels of biomarker cytokines can be increased by several different methods, including the administration of several compositions, the administration of one or more transgenic cells capable of expressing one or more recombinant cytokines, and any other methods that have the effect of increasing levels of biomarker cytokines in a patient.

[0103] In some embodiments, the present invention provides a patient with one or more pre-conditioning agents. The next step involves administering an effective dose of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. This includes methods for preconditioning patients who require T-cell therapy. The cytokines to be administered can be obtained by any method known in the art. For example, the cytokines may be isolated cytokines or recombinant cytokines. One or more doses of isolated cytokines or recombinant cytokines. It can be administered before T-cell therapy, after T-cell therapy, or in any combination thereof.

[0104] In one embodiment, a method for conditioning a patient requiring T-cell therapy involves administering to the patient one or more preliminary conditioning agents and one or more doses of IL-2. Including the floor. In some embodiments, the dose of IL-2 is at least about 10,000 IU / kg, at least The dose is approximately 50,000 IU / kg, at least approximately 100,000 IU / kg, at least approximately 200,000 IU / kg, at least approximately 400,000 IU / kg, at least approximately 600,000 IU / kg, at least approximately 700,000 IU / kg, at least approximately 800,000 IU / kg, or at least approximately 1,000,000 IU / kg. In one embodiment, the dose of IL-2 is small At least approximately 700,000 IU / kg. In a particular embodiment, the dose of IL-2 is approximately 720,000 IU / kg. In some embodiments, IL-2 is administered up to 15 doses or 8 times until toxicity prevents further doses. It is administered to the patient every so often.

[0105] Patients who have received one or more preliminary conditioning agents and / or T-cell therapy. In the serum, various cytokines can be concentrated. In some embodiments, patients after the administration of one or more preconditioning agents and / or T-cell therapy , interleukin (IL)15, IL-7, IL-10, IL-5, IL-8, IL-1, IL-1b, IL-2, IL-3, IL-4, IL-6, IL-9, IL-11, IL-12, IL-12p40, IL-12p70, IL-13, IL-14, IL-16, IL-17, IL-17a, IL-20, IL-21, granulocyte-macrophage colony-stimulating factor (GM-CS) F) Granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein 1 (MCP-1), MCP-4, gamma-inducible protein 10 (IP-10), placental growth factor (PLGF), soluble intercellular adhesion molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), C-reactive protein (CRP), vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D, macrophages Inflammatory protein 1β (MIP-1β, MIP-1b), leukemia suppressor factor (LIF), oncostatin M (OSM), interferon (IFN) alpha, IFN-beta, IFN-gamma, tumor necrosis factor (TNF) alpha Phosphorus, TNF-beta, CD154, lymphotoxin (LT) beta, 4-1BB ligand (4-1BBL), proliferation-inducing ligand (APRIL), CD70, CD153, CD178, glucocorticoid-inducing TNFR-related ligand (GITRL), tumor necrosis factor superfamily member 14 (TNFSF14), OX40L, TNF and ApoL-related Leukocyte expression ligand 1 (TALL-1), TNF-related apoptosis induction ligand (TRAIL), Chemocy CC motif ligand (CCL) 1, macrophage inflammatory protein 1 alpha (MIP-1α) This indicates increased serum concentrations of cytokines or pro-inflammatory factors selected from the group consisting of CCL3), CCL5, monocyte-specific chemokine 3 (MCP3 or CCL7), monocyte chemotactic protein 2 (MCP-2 or CCL8), CCL13, thymic and activation regulatory chemokines (TARC or CCL17), CCL22, FGF2, eotaxin, MDC, grandin A, grandin B, perforin, SAA, MCP-4, and any combination thereof. In some embodiments, cyclophosphamide and fludarabine are present. After administration, patients show elevated serum levels of IL-15 and / or IP-10. In this embodiment, after administration of cyclophosphamide and fludarabine, the patient exhibits decreased serum levels of perforin.

[0106] Pre-conditioning agent The present invention provides one or more preconditioning agents, including IL-15, IL-7, and MCP-1 A small number of CRP, PLGF, IP-10, and / or any combination thereof are selected from the group. At the very least, any ability to increase serum levels of one additional cytokine. It may be a preliminary conditioning agent. For example, one or more preliminary conditioning agents The alkylating agent may contain an alkylating agent. In certain embodiments, the alkylating agent may be melphalan, chlorambucil, cyclophosphamide, mechloretamine, mustine (HN2), The group can be selected from uramustine, uracil mustard, melphalan, chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, streptozosine, alkyl sulfonates, busulfan, thiotepa or its analogues, any analogue thereof or its functional derivatives, and any combination thereof. In a particular embodiment, one or more preconditioning agents include cyclophosphamide.

[0107] Cyclophosphamide (ENDOXAN®, CYTOXAN®, PROCYTOX®) Cyclophosphamide (Trademark), NEOSAR (Registered Trademark), REVIMMUNE (Registered Trademark), and CYCLOBLASTIN (Registered Trademark) are nitrogen mustard-derived alkylating agents with potent immunosuppressive activity. Cyclophosphamide acts as an anti-cancer agent and is used to treat various types of cancer, including lymphoma, multiple myeloma, leukemia, mycosis fungoides, neuroblastoma, ovarian cancer, eye cancer, and breast cancer, as well as autoimmune disorders.

[0108] Cyclophosphamide is converted to acrolein and phosphoramide in the liver after administration to the patient. Together, these metabolites crosslink DNA in both resting and dividing cells by adding alkyl groups to the nitrogen atom at position 7 of the imidazole ring of the guanine base of DNA. As a result, DNA replication is inhibited, leading to cell death.

[0109] In another embodiment, one or more preconditioning agents include platinum-based chemotherapeutic agents. In a particular embodiment, the platinum-based chemotherapeutic agent is selected from the group consisting of platinum, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, procarbazine, altretamine, triazene, dacarbazine, mitozolomide, temozolomide, dacarbazine, temozolomide, any analogue or functional derivative thereof, and any combination thereof.

[0110] In another embodiment, one or more preconditioning agents include purine analogs. This is possible. In a particular embodiment, the purine analog is selected from the group consisting of azathioprine, 6-mercaptopurine, mercaptopurine, thiopurine, thioguanine, fludarabine, pentostatin, cladribine, any analog or functional derivative thereof, and any combination thereof. In one embodiment, one or more preconditioning agents are fluda Contains rabine.

[0111] Fludarabine phosphate (FLUDARA®) is a sugar molecule with ribose or deoxygenated sugar. Fludarabine is a synthetic purine nucleoside that differs from physiological nucleosides in that it contains arabinose instead of siribose. It acts as a purine antagonist and is used to treat various types of hematological malignancies, including various lymphomas and leukemias.

[0112] After being administered to a patient, fludarabine is rapidly dephosphorylated to 2-fluoro-ara-A and then phosphorylated intracellularly by deoxycytidine kinase to the active triphosphate ester 2-fluoro-ara -ATP. This metabolite then inhibits DNA replication by inhibiting DNA synthesis, presumably by inhibiting DNA polymerase alpha, ribo nucleotide reductase, and DNA primase. As a result, administration of fludarabine leads to increased cell death in dividing cells .

[0113] In some embodiments, one or more preconditioning agents can include cyclophospha mide and purine analogs. The purine analogs can be selected from the group consisting of azathioprine, 6-mercaptopurine, mercaptopurine, thiopurine, thioguanine, fludarabine, pentostatin, cladribine, any analog or functional derivative thereof, and any combination thereof. In a particular embodiment, one or more preco nditioning agents include cyclophosphamide and pentostatin. In a particular embodiment, one or more preconditioning agents include cyclophosphamide and fluda rabine.

[0114] In certain embodiments, a first dose of one or more preconditioning agents is administered to the patient. For example, in some embodiments, the first dose of cyclophosphamide is from about 300 mg / m 2 / day to about 2000 mg / m 2 / day. In another embodiment, the first dose of cyclophosphamide is higher than 300 mg / m 2 / day and less than 2000 mg / m 2 / day. In other embodiments, the dose of cyclophosphamide is from about 350 mg / m 2 / day to about 2000 mg / m 2 / day, at least about 400 mg / m 2 / day~about 2000mg / m 2 / day, about 450mg / m 2 / day~about 2000mg / m 2 / day, about 500mg / m 2 / day~about 2000mg / m 2 / day, about 550mg / m 2 / day~about 2000mg / m 2 / day, or approximately 600 mg / m² 2 / day~about 2000mg / m 2 In another embodiment, the dose of cyclophosphamide is approximately 350 mg / m². 2 / day~about 1500mg / m 2 / day, about 350mg / m 2 / day~about 1000mg / m 2 / day, about 400mg / m 2 / day~about 900mg / m 2 / day, about 450mg / m 2 / day~about 800mg / m 2 / day, about 450mg / m 2 / day~about 700mg / m 2 / day, about 500mg / m 2 / day~about 600mg / m 2 / day, or approximately 300 mg / m² 2 / day~about 500mg / m 2 In another aspect, the dose of cyclophosphamide is approximately 350 mg / m². 2 / day, about 400mg / m 2 / day, about 450mg / m 2 / day, about 500mg / m 2 / day, about 550mg / m 2 / day, about 600mg / m 2 / day, approximately 650mg / m 2 / day, about 700mg / m 2 / day, Approximately 800mg / m 2 / day, approximately 900mg / m 2 / day, or approximately 1000 mg / m² 2 / day

[0115] In another embodiment, the first dose of cyclophosphamide is approximately 200 mg / m². 2 / day ~ approx. 3000mg / m 2 / day. In another embodiment, the first dose of cyclophosphamide is 200 mg / m². 2 Higher than / day, 3000mg / m² 2 The dose is less than 200 mg / m². In other embodiments, the dose of cyclophosphamide is approximately 200 mg / m². 2 / day ~ approx. 3000mg / m 2 / day, about 300mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 400mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 500mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 600mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 700mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 800mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 900mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 1000mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1100mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 1200mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1300mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1400mg / m 2 / day ~ approx. 3000mg / m 2 / day, about 1500mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1600mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1700mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1800mg / m 2 / day ~ approx. 3000mg / m 2 / day, approximately 1900mg / m 2 / day to approximately 3000 mg / m 2 / day, approximately 2000 mg / m 2 / day to approximately 3000 mg / m 2 / day, approximately 200 mg / m 2 / day to approximately 2900 mg / m 2 / day, approximately 400 mg / m 2 / day to approximately 2800 mg / m 2 / day, approximately 500 mg / m 2 / day to approximately 2700 mg / m 2 / day, approximately 600 mg / m 2 / day to approximately 2600 mg / m 2 / day, approximately 700 mg / m 2 / day to approximately 2500 mg / m 2 / day, approximately 800 mg / m 2 / day to approximately 2400 mg / m 2 / day, approximately 900 mg / m 2 / day to approximately 2350 mg / m 2 / day, approximately 1000 mg / m 2 / day to approximately 2300 mg / m 2 / day, approximately 1100 mg / m 2 / day to approximately 2250 mg / m 2 / day, or approximately 1110 mg / m 2 / day to approximately 2220 mg / m 2 / day. In some embodiments, the first dose of cyclophosphamide is 200 mg / m 2 / day. In another embodiment, the first dose of cyclophosphamide is 300 mg / m 2 / day. In another embodiment, the first dose of cyclophosphamide is 500 mg / m 2 / day.

[0116] In some embodiments, the first dose of fludarabine is approximately 20 mg / m 2 / day to approximately 900 mg / m 2 / day. In some embodiments, the dose of fludarabine is higher than 30 mg / m 2 / day and less than 900 mg / m 2 / day In some embodiments, the dose of fludarabine is approximately 35 mg / m 2 / day to approximately 900 mg / m2 / day, about 40mg / m 2 / day~about 900mg / m 2 / day, about 45mg / m 2 / day~about 900mg / m 2 / day, about 50mg / m 2 / day ~ approximately 90 0 mg / m² 2 / day, about 55mg / m 2 / day~about 900mg / m 2 / day, or approximately 60mg / m² 2 / day~about 900mg / m 2 / day In some embodiments, the dose of fludarabine is approximately 35 mg / m². 2 / day~about 900mg / m 2 / day, about 35mg / m 2 / day~about 800mg / m 2 / day, about 35mg / m 2 / day~about 700mg / m 2 / day, about 35mg / m 2 / day~about 600mg / m 2 / day, about 35mg / m 2 / day~about 500mg / m 2 / day, about 35mg / m 2 / day ~ approx. 400mg / m 2 / day, about 35mg / m 2 / day ~ approx. 300mg / m 2 / day, about 35mg / m 2 / day~about 200mg / m 2 / day, about 35mg / m 2 / day~about 100mg / m 2 / day, about 40mg / m 2 / day~ Approximately 90mg / m 2 / day, about 45mg / m 2 / day~about 80mg / m 2 / day, about 45mg / m 2 / day~about 70mg / m 2 / day, or approximately 50 mg / m² 2 / day~about 60mg / m 2 The dose is approximately 20 mg / m². In some embodiments, the dose of fludarabine is approximately 20 mg / m². 2 / day, about 25mg / m2 / day, about 30mg / m 2 / day, about 35mg / m 2 / day, about 40mg / m 2 / day, about 45mg / m 2 / day, about 50mg / m 2 / day, about 55mg / m 2 / day, about 60mg / m 2 / day, about 65mg / m 2 / day, about 70mg / m 2 / day, about 75mg / m 2 / day, about 80mg / m 2 / day, about 85mg / m 2 / day, approximately 90mg / m 2 / day, approximately 95mg / m 2 / day, about 100mg / m 2 / day, about 200mg / m 2 / day, or approximately 300 mg / m² 2 The dose is approximately 20 mg / m². In some embodiments, the dose of fludarabine is approximately 20 mg / m². 2 / day, about 25mg / m 2 / day, about 30mg / m 2 / day, about 35mg / m 2 / day, about 40mg / m 2 / day, about 45mg / m 2 / day, Approximately 50mg / m 2 / day, about 55mg / m 2 / day, about 60mg / m 2 / day, about 65mg / m 2 / day, about 70mg / m 2 / day, about 75mg / m 2 / day, about 80mg / m 2 / day, about 85mg / m 2 / day, approximately 90mg / m 2 / day, approximately 95mg / m 2 / day, or approximately 100 mg / m² 2 / day In other embodiments, the dose of fludarabine is approximately 110 mg / m². 2 / day, 120mg / m 2 / day, 130mg / m 2 / day, 140mg / m 2 / day, 150mg / m 2 / day, 160mg / m2 / day, 170mg / m 2 / day, 180mg / m 2 / day, or 190 mg / m² 2 The dose is approximately 210 mg / m². In some embodiments, the dose of fludarabine is approximately 210 mg / m². 2 / day, 220mg / m 2 / day, 230mg / m 2 / day, 240mg / m 2 / day, 250mg / m 2 / day, 260mg / m 2 / day, 270mg / m 2 / day, 280mg / m 2 / 290 mg / m² per day, or 290 mg / m² 2 The dose is approximately 20 mg / m². In one particular embodiment, the dose of fludarabine is approximately 20 mg / m². 2 / day In one particular embodiment, the dose of fludarabine is approximately 25 mg / m². 2 In another embodiment, the dose of fludarabine is approximately 30 mg / m². 2 / day In another embodiment, the dose of fludarabine is approximately 60 mg / m². 2 / day

[0117] In some embodiments, the dose of cyclophosphamide is 100 mg / m². 2 / day (or 110mg / m²) 2 / day , 120 mg / m² 2 / day, 130mg / m 2 / day, or 140mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0118] In some embodiments, the dose of cyclophosphamide is 150 mg / m². 2 / day (or 160mg / m²) 2 / day , 170 mg / m² 2 / day, 180mg / m 2 / day, or 190mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0119] In some embodiments, the dose of cyclophosphamide is approximately 200 mg / m². 2 / day (or 210 mg / m²) 2 / day, 220mg / m 2 / day, 230mg / m 2 / day, or 240mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day, 40mg / m 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0120] In some embodiments, the dose of cyclophosphamide is 250 mg / m². 2 / day (or 260mg / m²) 2 / day , 270 mg / m² 2 / day, 280mg / m 2 / day, or 290mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0121] In some embodiments, the dose of cyclophosphamide is 300 mg / m². 2 / day (or 310mg / m²) 2 / day , 320 mg / m² 2 / day, 330mg / m 2 / day, or 340mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m².2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0122] In some embodiments, the dose of cyclophosphamide is 350 mg / m². 2 / day (or 360mg / m²) 2 / day , 370 mg / m² 2 / day, 380mg / m 2 / day, or 390mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0123] In some embodiments, the dose of cyclophosphamide is 400 mg / m². 2 / day (or 410mg / m²)2 / day , 420 mg / m² 2 / day, 430mg / m 2 / day, or 440mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0124] In some embodiments, the dose of cyclophosphamide is 450 mg / m². 2 / day (or 460mg / m²) 2 / day , 470 mg / m² 2 / day, 480mg / m 2 / day, or 490mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0125] In some embodiments, the dose of cyclophosphamide is 500 mg / m². 2 / day (or 510mg / m²) 2 / day , 520 mg / m² 2 / day, 530mg / m 2 / day, or 540mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0126] In some embodiments, the dose of cyclophosphamide is 550 mg / m². 2 / day (or 560mg / m²) 2 / day 570 mg / m² 2 / day, 580mg / m 2 / day, or 590mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0127] In some embodiments, the dose of cyclophosphamide is 600 mg / m². 2 / day (or 610mg / m²) 2 / day , 620 mg / m² 2 / day, 630mg / m 2 / day, or 640mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0128] In some embodiments, the dose of cyclophosphamide is 650 mg / m². 2 / day (or 660mg / m²) 2 / day , 670 mg / m² 2 / day, 680mg / m 2 / day, or 690mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0129] In some embodiments, the dose of cyclophosphamide is 700 mg / m². 2 / day (or 710mg / m²) 2 / day 720 mg / m² 2 / day, 730mg / m 2 / day, or 740mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0130] In some embodiments, the dose of cyclophosphamide is 750 mg / m². 2 / day (or 760mg / m²) 2 / day 770 mg / m² 2 / day, 780mg / m2 / day, or 790mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0131] In some embodiments, the dose of cyclophosphamide is 800 mg / m². 2 / day (or 810 mg / m²) 2 / day , 820 mg / m² 2 / day, 830mg / m 2 / day, or 840mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0132] In some embodiments, the dose of cyclophosphamide is 850 mg / m². 2 / day (or 860mg / m²) 2 / day , 870 mg / m² 2 / day, 880mg / m 2 / day, or 890mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0133] In some embodiments, the dose of cyclophosphamide is 900 mg / m². 2 / day (or 910mg / m²) 2 / day 920 mg / m² 2 / day, 930mg / m 2 / day, or 940mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0134] In some embodiments, the dose of cyclophosphamide is 950 mg / m². 2 / day (or 960mg / m²) 2 / day 970 mg / m² 2 / day, 980mg / m 2 / day, or 990mg / m² 2 The dosage is 5 mg / m² (per day), and the dose of fludarabine is 5 mg / m². 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day , 40 mg / m² 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0135] In some embodiments, the dose of cyclophosphamide is 1000 mg / m². 2 / day (or 1010 mg / m²) 2 / day, 1020mg / m 2 / day, 1030mg / m 2 / day, or 1040mg / m² 2 The dosage of fludarabine is ( / day). It is 5 mg / m² 2 / day, 10mg / m 2 / day, 15mg / m 2 / day, 20mg / m 2 / day, 25mg / m 2 / day, 30mg / m 2 / day, 35mg / m 2 / day, 40mg / m 2 / day, 45mg / m 2 / day, 50mg / m 2 / day, 55mg / m 2 / day, 60mg / m 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, or 75mg / m² 2 / day

[0136] In other embodiments, the dose of cyclophosphamide is 100 mg / m². 2 / day~650mg / m 2 The daily dose is 10 mg / m² of fludarabine. 2 / day~50mg / m 2 In another embodiment, the dose of cyclophosphamide is 150 mg / m². 2 / day~600mg / m 2 The daily dose is 20 mg / m², and the fludarabine dose is 20 mg / m². 2 / day~50mg / m 2 In another embodiment, the dose of cyclophosphamide is 200 mg / m². 2 / day~550mg / m 2 The daily dose is 20 mg / m², and the fludarabine dose is 20 mg / m². 2 / day~40mg / m 2 In another embodiment, the dose of cyclophosphamide is 250 mg / m². 2 / day~550mg / m 2 The daily dose is 15 mg / m² of fludarabine. 2 / day~45mg / m 2 / day

[0137] In a particular embodiment, the dose of cyclophosphamide is 1000 mg / m². 2 The daily dose is 60 mg / m² of fludarabine. 2 / day, 65mg / m 2 / day, 70mg / m 2 / day, 75mg / m 2 / day, 80mg / m 2 / day, 85mg / m2 / day, 90mg / m 2 / day, 95mg / m 2 / day, 100mg / m 2 / day, 105mg / m 2 / day, 110mg / m 2 / day, 115mg / m 2 / day, 120mg / m 2 / day, 125mg / m 2 / day, 130mg / m 2 / day, 135mg / m 2 / day, 140mg / m 2 / day, 145mg / m 2 / day, 150mg / m 2 / day, 155mg / m 2 / day, 160mg / m 2 / day, 165mg / m 2 / day, 170mg / m 2 / day, 175mg / m 2 / day, 180mg / m 2 / day, 185mg / m 2 / day, 190mg / m 2 / day, 195mg / m 2 / day, 200mg / m 2 / day, 205mg / m 2 / day, 210mg / m 2 / day, 215mg / m 2 / day, 220mg / m 2 / day, 225mg / m 2 / day, 230mg / m 2 / day, 235mg / m 2 / day, 240mg / m 2 / day, 245 mg / m 2 / day, or 250mg / m² 2 / day

[0138] In one embodiment, the dose of cyclophosphamide is approximately 500 mg / m². 2 The dosage of fludarabine is per day. Approximately 60 mg / m² 2 In another embodiment, the dose of cyclophosphamide is approximately 300 mg / m². 2 / day The dosage of fludarabine is approximately 30 mg / m². 2 In another embodiment, the dose of cyclophosphamide is approximately 200 mg / m². 2 The daily dose is approximately 20 mg / m² of fludarabine. 2 / day. In another aspect, The dosage of cyclophosphamide is approximately 200 mg / m². 2 The daily dose of fludarabine is approximately 30 mg / m². 2 / day Yes. In another embodiment, the dose of cyclophosphamide is approximately 500 mg / m². 2 / day and fludarabine The dosage is approximately 30 mg / m². 2 In another embodiment, the dose of cyclophosphamide is approximately 300 mg / m². 2 / The daily dose of fludarabine is approximately 60 mg / m². 2 In another aspect, the dose of cyclophosphamide is approximately 500 mg / m². 2 The daily dose is approximately 60 mg / m² of fludarabine. 2 / day So, the dosage of cyclophosphamide is approximately 1110 mg / m². 2 The daily dose is approximately 25 mg / m² of fludarabine. 2 In another embodiment, the dose of cyclophosphamide is approximately 2220 mg / m². 2 The daily dose is approximately 25 mg / m² of fludarabine. 2 The daily dose of cyclophosphamide and fludarabine indicates elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, such as elevated serum levels of IL-15, IP-10, and / or IL-7, or decreased serum levels of perforin and / or MIP-1b. This indicates the level of purity. In another embodiment, the dose of cyclophosphamide is approximately 60 mg / kg / day, and the dose of fludarabine is approximately 25 mg / m². 2The daily dose is used to measure the levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, after administration of cyclophosphamide and fludarabine. The serum levels of IL-15, IP-10, and / or IL-7 are elevated, or serum levels of perforin and / or MIP-1b are reduced. The levels are indicated. In another embodiment, the dose of cyclophosphamide is approximately 30 mg / kg / day, and the dose of fludarabine is approximately 25 mg / m². 2 The dosage is per day. In a particular embodiment, cyclophosphamide is administered before, after, or concurrently with fludarabine. In a particular embodiment, cyclophosphamide is administered before fludarabine.

[0139] The timing of administering one or more pre-conditioning agents is crucial for maximizing their effectiveness. It can be adjusted to do so. In a particular embodiment, one or more preconditioners The conditioning agent comprises two or more preconditioning agents. The two or more preconditioning agents can be administered simultaneously or sequentially. In a particular embodiment, the first preconditioning agent Conditioning agents, such as cyclophosphamide, are a second preliminary conditioning agent. For example, it may be administered to the patient before or after fludarabine.

[0140] The doses of cyclophosphamide and fludarabine can be increased or decreased together or independently. For example, the dose of cyclophosphamide can be increased while the dose of fludarabine is decreased, and the dose of cyclophosphamide can be decreased while the dose of fludarabine is increased. Alternatively, the doses of both cyclophosphamide and fludarabine can be increased or decreased together. In some embodiments, the dose of cyclophosphamide is 300 mg / m². 2 The daily dose is 20 mg / m² of fludarabine. 2 / day Yes. In another embodiment, the dose of cyclophosphamide is 300 mg / m². 2 / day, fludarabine The dosage is 30 mg / m². 2 In another embodiment, the dose of cyclophosphamide is 300 mg / m². 2 / day The dosage of fludarabine is 60 mg / m². 2 In another embodiment, the dose of cyclophosphamide is 500 mg / m². 2 The daily dose is 20 mg / m² of fludarabine. 2 / day. In other forms, cyclo The dosage of phosphamide is 500 mg / m². 2 The daily dose is 30 mg / m² of fludarabine. 2 / day. In this configuration, the dose of cyclophosphamide is 500 mg / m². 2 The daily dose is 60 mg / m² of fludarabine. 2 In another embodiment, the dose of cyclophosphamide is 200 mg / m². 2 The daily dose is 20 mg / m² of fludarabine. 2 In another embodiment, the dose of cyclophosphamide is 200 mg / m². 2 The daily dose is 30 mg / m² of fludarabine. 2 / day. In other embodiments, cyclophosphamide The dosage is 200 mg / m². 2 The daily dose is 60 mg / m² of fludarabine. 2 / day

[0141] As described herein, the day on which T-cell therapy is administered is referred to as Day 0. A number of pre-conditioning agents can be administered at any time before T-cell therapy is performed. In some embodiments, administration of one or more preconditioning agents begins at least 7 days, at least 6 days, at least 5 days, at least 4 days, at least 3 days, at least 2 days, or at least 1 day before the administration of T-cell therapy. In other embodiments, administration of one or more preconditioning agents begins at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, or at least 12 days before the administration of T-cell therapy. Both begin 13 days prior, or at least 14 days prior. In one embodiment, one or more backup units The administration of conditioning agents begins approximately 7 days before the administration of T-cell therapy. In another embodiment, one The administration of multiple pre-conditioning agents begins approximately 5 days before the start of T-cell therapy.

[0142] In one embodiment, the administration of the first pre-conditioning agent begins approximately 7 days before the implementation of T-cell therapy. Furthermore, the administration of the second pre-conditioning agent begins approximately 5 days before the start of T-cell therapy. In one particular embodiment, the first pre-conditioning agent is administered to the patient for two days, approximately seven or six days before the administration of T-cell therapy. In another embodiment, the second pre-conditioning agent is administered to the patient for five days, approximately five, four, three, two, and one day before the administration of T-cell therapy. In yet another embodiment... The first preliminary conditioning agent is administered to the patient approximately 5, 4, and 3 days before the implementation of T-cell therapy. It is administered over a three-day period prior to the treatment.

[0143] In a particular embodiment, cyclophosphamide administration begins approximately 7 days before the administration of T-cell therapy, and purine analogues (e.g., fludarabine or pentostatin) are administered before the administration of T-cell therapy. Administration begins approximately 5 days before the administration. In another embodiment, cyclophosphamide administration begins approximately 5 days before the administration of T-cell therapy, and purine analogs (e.g., fludarabine or pentostatin) are administered. This begins approximately 5 days before the administration of T-cell therapy.

[0144] The timing of administration for each component can be adjusted to maximize its effect. Generally, one or more pre-conditioning agents can be administered daily. In some embodiments, one or more pre-conditioning agents are administered daily for about 2, 3, 4, 5, 6, or 7 days. In some embodiments, one or more pre-conditioning agents are administered daily for at least 1, 2, 3, 4, 5, 6, or 7 days. It can be administered daily. In a particular embodiment, one or more preconditioning agents The medication is administered daily for approximately three days.

[0145] As described herein, the day on which T-cell therapy is administered to a patient is referred to as day 0. In some embodiments, one or more preconditioning agents, such as cyclophosphamide, are administered. It is administered to the patient 7 and 6 days prior to day 0 (i.e., day -7 and day -6). In this embodiment, one or more preconditioning agents, for example cyclophosphamide, It is administered to the patient on day -5, day -4, and day -3. In some embodiments, one or Multiple pre-conditioning agents, such as fludarabine, are administered to the patient on days -5, -4, -3, -2, and -1. In other embodiments, one or more pre-conditioning agents A conditioning agent, such as fludarabine, is administered to the patient on days -5, -4, and -3.

[0146] One or more pre-conditioning agents, e.g., cyclophosphamide and flu Darabine can be administered on the same or different days. If cyclophosphamide and fludarabine are administered on the same day, the dose of cyclophosphamide should be equal to the dose of fludarabine. It can be administered either before or after. In one embodiment, doses of cyclophosphamide are administered to the patient on days -7 and -6, and doses of fludarabine are administered to the patient on days -5, -4, -3, -2, and -1. In another embodiment, doses of cyclophosphamide are administered to the patient on days -5, -4, and -3, and doses of fludarabine are administered to the patient on days -5, -4, and -3.

[0147] In certain embodiments, one or more preconditioning agents, for example, cyclophosphates Sphamide and fludarabine can be administered simultaneously or sequentially. In one embodiment, cyclophosphamide is administered to the patient before fludarabine. In another embodiment, cyclophosphamide is administered to the patient after fludarabine.

[0148] One or more pre-conditioning agents, administered intravenously (IV) or orally, as per any medical procedure. It can be administered by route. In some embodiments, one or more preconceived A conditioning agent, such as cyclophosphamide, is administered intravenously over approximately 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 90 minutes, and 120 minutes. In some embodiments, one or more preconditioning agents, such as fludarabine, are administered intravenously over approximately 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 90 minutes, and 120 minutes.

[0149] In certain embodiments, T-cell therapy is administered to the patient after administration of one or more preconditioning agents, such as cyclophosphamide and fludarabine. In some embodiments, T-cell therapy includes adoptive cell therapy. In certain embodiments, adoptive cell therapy is used to treat tumors. Therapies are selected from tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, modified autologous cell therapy (eACT), and allogeneic T cell transplantation. In certain embodiments, eACT includes the administration of modified antigen-specific chimeric antigen receptor (CAR)-positive (+) T cells. In other embodiments, eACT includes the administration of modified antigen-specific T cell receptor (TCR)-positive (+) T cells. In some embodiments, modified T cells treat the tumor in the patient.

[0150] Various other interventions may be included in the methods described herein. For example, preconditioning agents, such as cyclophosphamide and fludarabine, are well known to cause adverse events in patients after administration. It is also within the scope of the invention that compositions may be administered to patients to reduce some of these adverse events. In some embodiments, the method further includes the step of administering saline to the patient. Saline is one of several or before or after the administration of multiple pre-conditioning agents, or even one of them It can be administered to the patient both before and after the administration of one or more preconditioning agents. In certain embodiments, saline solution is administered to one or more preconditioning agents. It is administered simultaneously with the suppository. In a particular embodiment, saline solution is administered one or more times on each infusion day. Before administration of multiple pre-conditioning agents and one or more pre-conditioning agents It is administered to the patient after the administration of the drug.

[0151] Physiological saline can be administered to the patient by any route, including intravenous or oral administration. In some embodiments, this method includes administering approximately 0.1 L, 0.2 L, 0.3 L, 0.4 L, 0.5 L, 0.6 L, 0.7 L, 0.8 L, 0.9 L, 1 L, 1.1 L, 1.2 L, 1.3 L, 1.4 L, 1.5 L, 1.6 L, 1.7 L, 1.8 L, 1.9 L, or 2.0 L of physiological saline. The NaCl content of the physiological saline is approximately 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, and 0.7%. It can be dissolved to a concentration of approximately 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or 2.0%. In one embodiment, the method includes the step of administering 1.0 L of 0.9% NaCl saline to the patient. In a particular embodiment, the method includes the administration of one or more preconditioning agents on each infusion day. Before administration and after administration of one or more pre-conditioning agents, the patient received 0.9% NaCl physiological This includes the step of administering 1.0 L of saline solution.

[0152] Furthermore, adjuvants and excipients can also be administered to the patient. For example, mesna(2-sulfur Sodium 2-sulfanylthanesulfonate is an adjuvant that acts as an antidote to prevent hemorrhagic cystitis and hematuria that may occur after treatment with cyclophosphamide. Cyclophosphamide can be converted in vivo to uremic metabolites such as acrolein. Mesna helps detoxify these metabolites through the reaction of its sulfhydryl group with its vinyl group. Mesna also increases the urinary excretion of cysteine. In certain embodiments, this method further includes the step of administering mesna to the patient. Mesna is administered before the administration of cyclophosphamide and / or fludarabine. After administration of cyclophosphamide and / or fludarabine, or after administration of cyclophosphamide It can be administered both before and after the administration of fludarabine. In one embodiment Mesna is administered intravenously or orally (via the mouth). For example, oral mesna can be given together with oral cyclophosphamide.

[0153] In addition, in the methods described herein, exogenous cytokines may also be administered to the patient. As described above, reducing the number of endogenous lymphocytes promotes the proliferation and activity of adoptively transplanted T cells. It is hypothesized that this method increases the bioavailability of endogenous molecules such as cytokines, which may be favorable for sexualization and transport. Therefore, various cytokines can be administered to the patient. In one embodiment, the method further comprises the step of administering one or more doses of IL-2, IL-15, IL-7, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof. In a particular embodiment, the method further comprises one or more doses This includes a step of administering a certain amount of IL-2. The dose of IL-2 is at least about 10,000 IU / kg, at least It may be approximately 50,000 IU / kg, at least approximately 100,000 IU / kg, at least approximately 200,000 IU / kg, at least approximately 400,000 IU / kg, at least approximately 600,000 IU / kg, at least approximately 700,000 IU / kg, at least approximately 800,000 IU / kg, or at least approximately 1,000,000 IU / kg.

[0154] T cell therapy This invention increases serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. To administer one or more pre-conditioning agents to the patient that have the ability to enhance the condition Therefore, in patients suitable for T-cell therapy, including the stage of preconditioning the patient A method for treating cancer, provided that the patient is treated with T-cell therapy when the patient exhibits elevated serum levels of IL-15, IL-7, and at least one additional cytokine. The preliminary conditioning method generally involves homeostatic proliferation, activation, and Various different T-cell therapies, as described herein, help to modify the immune environment by inducing IL-15, IL-7, and at least one additional cytokine, which may be favorable for transport. Benefits can be obtained from this conditioning method. Those skilled in the art will understand that this method can be applied to any method of treating a patient, which includes the step of administering one or more T cells to the patient.

[0155] For example, the methods described herein include, but are not limited to, tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell immunotherapy, etc. The effectiveness of T cell therapy can be enhanced by adoptive T cell therapy, which is selected from a group consisting of cytotherapy, manipulated autologous cell therapy (eACT), allogeneic T cell transplantation, non-T cell transplantation, and any combination thereof. Adoptive T cell therapy has the ability to recognize and bind to tumor cells. This broadly includes any method of selecting, in vitro enriching, and administering a patient's own T cells or allogeneic T cells to the patient. TIL immunotherapy has the ability to invade tumor tissue. In adoptive T-cell therapy, lymphocytes are isolated, enriched in vitro, and administered to the patient. It is a type. TIL cells can be either autologous or allogeneic. Autologous cell therapy is used by patients. Adoptive T-cell therapy involves the steps of isolating T cells capable of targeting tumor cells from a patient, enriching the T cells in vitro, and administering the T cells back to the same patient. Allogeneic T-cell transplantation involves the transplantation of ex vivo-grown native T cells or genetically modified T cells. This may include a fragment. The modified autologous cell therapy described in more detail above is adoptive T-cell therapy in which the patient's own lymphocytes are isolated, genetically modified to express tumor-targeting molecules, proliferated in vitro, and administered back to the patient. Non-T cell transplantation may, without limitation, include autologous or allogeneic therapies using non-T cells such as natural killer (NK) cells.

[0156] In a particular embodiment, the T-cell therapy of the present invention is a modified autologous cell therapy (eACT®). In some embodiments, this method may include a step of collecting blood cells from a patient before administering one or more preconditioning agents. The isolated blood cells (e.g., T cells) may then be engineered to express chimeric antigen receptors ("engineered CAR T cells") or T cell receptors ("engineered TCR T cells"). In some embodiments, T cell therapy may include This includes engineered CAR T cell therapy or engineered TCR T cell therapy. In certain embodiments, engineered CAR T cells Alternatively, manipulated TCR T cells are administered to patients after receiving one or more preconditioning agents. It is administered to the patient. In some embodiments, engineered CAR T cells or engineered TCR T cells treat tumors in the patient. In some embodiments, engineered CAR T cells reduce tumor size. In other embodiments, Manipulated TCR T cells reduce tumor size.

[0157] In one embodiment, T cells are engineered to express chimeric antigen receptors. The protore receptor may contain a binding molecule to the tumor antigen. The binding molecule may be an antibody or its antigen-binding molecule. For example, the antigen-binding molecule can be selected from scFv, Fab, Fab', Fv, F(ab')2, and dAb, as well as any fragment or combination thereof. ru.

[0158] Chimeric antigen receptors may further contain hinge regions. These hinge regions may originate from the hinge regions of IgG1, IgG2, IgG3, IgG4, IgA, IgD, IgE, IgM, CD28, or CD8 alpha. This is possible. In a particular embodiment, the hinge region originates from the hinge region of IgG4.

[0159] A chimeric antigen receptor may also include a transmembrane domain. The transmembrane domain may be a transmembrane domain of any transmembrane molecule that is a co-receptor on an immune cell, or a transmembrane domain of a member of the immunoglobulin superfamily. In certain embodiments, the transmembrane domain is derived from a CD28, CD8 alpha, CD4, or CD19 transmembrane domain. In a particular embodiment, the transmembrane domain includes a domain derived from a CD28 transmembrane domain.

[0160] The chimeric antigen receptor further comprises one or more co-stimulatory signaling regions. This is possible. For example, the co-stimulatory signaling region includes CD28, OX-40, 4-1BB, CD27, and inductive T cells. This may be a vesicular co-stimulatory molecule (ICOS), CD3 gamma, CD3 delta, CD3 epsilon, CD247, Ig alpha (CD79a), or a signaling region of the Fc gamma receptor. In a particular embodiment, co-stimulatory The signaling region is the CD28 signaling region.

[0161] In one embodiment, the chimeric antigen receptor further comprises a CD3 zeta signaling domain. .

[0162] Chimeric antigen receptors can be engineered to target specific tumor antigens. In some embodiments, the tumor antigens are CD19, CD20, ROR1, CD22, carcinoembryonic antigen, ALF Afetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, variant p53, variant ras, HER2 / Neu, folate-binding protein, HIV-1 envelope HIV-1 envelope glycoproteins gpl20, gp41, GD2, CD123, CD33, CD138 CD23, CD30, CD56, c-Met, Mesothelin, GD3, HERV-K, IL-IIR Alpha, Kappa Chain, La The tumor antigen is selected from the following: useless chains, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 combinations, HER1-HER2 combinations, and any combination thereof. In a particular embodiment, the tumor antigen is CD19.

[0163] In another embodiment, T cell therapy includes the step of administering modified T cells expressing a T cell receptor ("modified TCR T cells") to a patient. The T cell receptor (TCR) may contain a binding molecule to a tumor antigen. In some embodiments, the tumor antigen may be CD19, CD20, ROR1, CD22, etc. The following are selected from the group consisting of fetal antigens, alpha-fetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, variant p53, variant ras, HER2 / Neu, folate-binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c-Met, mesothelin, GD3, HERV-K, IL-IIR alpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 combinations, HER1-HER2 combinations, and any combination thereof.

[0164] In one embodiment, the TCR includes a binding molecule to the viral oncogene. In a particular embodiment, The viral oncogenes are selected from human papillomavirus (HPV), Epstein-Barr virus (EBV), and human T-lymphotropic virus (HTLV).

[0165] In another embodiment, the TCR includes a binding molecule to the testis, placenta, or fetal tumor antigen. In a specific embodiment, the testicular, placental, or fetal tumor antigen is NY-ESO-1, synovial sarcoma X-ray. A group consisting of cyton 2 (SSX2), melanoma antigen (MAGE), and any combination thereof. It will be selected.

[0166] In another embodiment, the TCR includes a binding molecule for the series-specific antigen. In a particular embodiment, the system Cell-specific antigens are selected from a group consisting of melanoma antigen 1 (MART-1), gp100, prostate-specific antigen (PSA), prostate-specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), and any combination thereof, all of which are recognized by T cells.

[0167] In one embodiment, T cell therapy expresses a chimeric antigen receptor that binds to CD19, Furthermore, the modified CAR T cells include the CD28 costimulatory domain and the CD3-zeta signaling region. This includes the step of administering to the patient. In certain embodiments, T-cell therapy includes the step of administering KTE-C19 to the patient.

[0168] The T-cell therapy included in this invention involves transplanting T cells into the patient. The T cells are administered in a therapeutically effective dose. It can be administered. For example, a therapeutically effective amount of T cells, for example, manipulated CAR+ T cells or manipulated T cells. TCR+ T cells are at least about 10 4 pieces, at least about 10 5 pieces, at least about 10 6 few Approximately 10 7 pieces, at least about 10 8 pieces, at least about 10 9 one, or at least about 10 10 Individual It is possible. In another embodiment, a therapeutically effective amount of T cells, for example, engineered CAR+ T cells or engineered TCR+ T cells. The cells are about 10 4 pieces, about 10 5 pieces, about 10 6 pieces, about 10 7 10, or about 10 8 It is an individual. In a specific manner A therapeutically effective amount of T cells, such as engineered CAR+ T cells or engineered TCR+ T cells, is approximately 1 × 10⁶ 5 pieces / kg, approximately 2×10 5pieces / kg, approximately 3×10 5 pieces / kg, approximately 4×10 5 pieces / kg, approximately 5×10 5 pieces / kg, approximately 6×10 5 pieces / kg, Approximately 7×10 5 pieces / kg, approximately 8×10 5 pieces / kg, approximately 9×10 5 pieces / kg, approximately 1×10 6 pieces / kg, approximately 2×10 6 pieces / kg, approximately 3×10 6 pieces / kg, approximately 4×10 6 pieces / kg, approximately 5×10 6 pieces / kg, approximately 6×10 6 pieces / kg, approximately 7×10 6 pieces / kg, approximately 8×10 6 pieces / kg, approximately 9×10 6 pieces / kg, approximately 1×10 7 pieces / kg, approximately 2×10 7 pieces / kg, approximately 3×10 7 pieces / kg, approximately 4×10 7 pieces / kg, approximately 5×10 7 pieces / kg, approximately 6×10 7 pieces / kg, approximately 7×10 7 pieces / kg, approximately 8×10 7 pieces / kg, or approximately 9 x 10 7 pieces / kg In a particular aspect, the treatment of T cells, for example, manipulated CAR+ T cells or manipulated TCR+ T cells, is performed. The effective dose is approximately 2 × 10⁻⁶ 6 It is per kilogram.

[0169] In other embodiments, a therapeutically effective amount of T cells, for example, engineered CAR+ T cells or engineered TCR+ T cells, , about 1.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 2.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 3.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 4.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 5.0×105 Units / kg ~ Approximately 2 × 10 8 Units / kg Approximately 6.0 × 10 5 Units / kg ~ Approximately 2 × 10 8 Units / kg, approximately 7.0 × 10 5 Units / kg ~ Approximately 2 × 10 8 Units / kg, approximately 8.0 × 10 5 Units / kg ~ Approximately 2 × 10 8 Units / kg, approximately 9.0 × 10 5 Units / kg ~ Approximately 2 × 10 8 per kg, approximately 0.5 × 10 6 Units / kg ~ Approximately 2 × 10 8 Units / kg, approximately 2 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 3 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 4 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 5 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 6 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 7 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 8 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 9 × 10 6 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 1×10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 2 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 3 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 4 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 5 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 6 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 7 × 10 7 pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 8 × 10 7pcs / kg ~ approximately 9 x 10 7 Units / kg, approximately 2 × 10 6 8 pieces / kg ~ approximately 10 7 Units / kg, approximately 2 × 10 6 pcs / kg ~ approximately 7×10 7 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 6 × 10 7 Units / kg, approximately 2 × 10 6 5 pieces / kg ~ approximately 10 7 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 4 × 10 7 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 3 × 10 7 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 2 × 10 7 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 1×10 7 Units / kg, approximately 2 × 10 6 pcs / kg ~ approximately 9 x 10 6 Units / kg, approximately 2 × 10 6 8 pieces / kg ~ approximately 10 6 Units / kg, approximately 2 × 10 6 pcs / kg ~ approximately 7×10 6 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 6 × 10 6 Units / kg, approximately 2 × 10 6 5 pieces / kg ~ approximately 10 6 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 4 × 10 6 Units / kg, approximately 2 × 10 6 Units / kg ~ Approximately 3 × 10 6 Units / kg, approximately 3 × 10 6 8 pieces / kg ~ approximately 10 7 Units / kg, approximately 4 × 10 6 pcs / kg ~ approximately 7×10 7 Units / kg, approximately 5 × 10 6 Units / kg ~ Approximately 6 × 10 7 Units / kg, approximately 6 × 10 6 5 pieces / kg ~ approximately 10 7 Units / kg, approximately 7 × 10 6 Units / kg ~ Approximately 4 × 10 7 Units / kg, approximately 8 × 10 6 Units / kg ~ Approximately 3 × 10 7pieces / kg, or approximately 9 x 10 6 pieces / kg ~ approx. 2×10 7 The number is cells / kg. In one embodiment, the therapeutically effective amount of manipulated CAR T cells is approximately 0.8 × 10⁶ T cells. 6 pieces / kg ~ approx. 1.2×10 6 It is pcs / kg. In a particular embodiment, the therapeutically effective amount of CAR T The cells are 2.0 × 10 5 It is cells / kg. In a particular embodiment, the therapeutically effective amount of manipulated CAR T cells is 1.0 ×10 6 It is per kilogram.

[0170] Cancer treatment The methods of the present invention can be used to treat cancer in a subject, reduce tumor size, kill tumor cells, prevent tumor cell proliferation, prevent tumor growth, remove tumor from a patient, prevent tumor recurrence, prevent tumor metastasis, induce remission in a patient, or any combination thereof. In certain embodiments, the method induces a complete response. In other embodiments, the method induces a partial response.

[0171] Cancers that can be treated include non-angiogenic, substantially non-angiogenic, or angiogenic tumors. Cancers may also include solid or non-solid tumors. In certain embodiments, cancers include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, and T-cell adenomatous cancer. Large B-cell lymphoma (TCRBCL), mediastinal large B-cell lymphoma (PMBCL), non-Hodgkin lymphoma This includes cancers such as peritoneum, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, T-cell lymphoma, and asbestos-induced cancers. Tumors can be selected from cancer and tumors derived from combinations of the aforementioned cancers.

[0172] In one embodiment, this method can be used to treat tumors, in which case the tumor is lymphoma or leukemia. Lymphoma and leukemia are blood cancers that specifically affect lymphocytes. All white blood cells in the blood originate from a single type of pluripotent hematopoietic stem cell found in the bone marrow. These stem cells produce both myeloid progenitor cells and lymphoid progenitor cells, which then give rise to the various types of white blood cells found in the body. White blood cells that arise from myeloid progenitor cells include T lymphocytes (T cells), B lymphocytes (B cells), natural killer cells, and plasma cells. White blood cells that arise from lymphoid progenitor cells include megakaryocytes, mast cells, basophils, neutrophils, eosinophils, monocytes, and macrophages. Leukemia can affect one or more of these cell types in a patient.

[0173] Generally, lymphomas are classified into at least two subgroups: Hodgkin lymphoma and non-Hodgkin lymphoma. It can be classified as a digkin lymphoma. Non-Hodgkin lymphoma (NHL) involves B lymphocytes, T lymphocytes. It is a heterogeneous group of cancers that originate from lymphocytes or natural killer cells. In the United States, B Cellular lymphoma accounts for 80-85% of reported cases. In 2013, it was estimated that there were approximately 69,740 new cases of NHL and over 19,000 deaths associated with the disease. (Non-Hodgkin's lymphoma) Carbuncle is the most prevalent hematological malignancy, ranking seventh among new cancer sites in men and women, and accounting for 4% of all new cancer cases and 3% of cancer-related deaths.

[0174] Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of NHL, accounting for approximately 30% of all NHL cases. In the United States, approximately 22,000 new cases of DLBCL are diagnosed each year. This is classified as an aggressive lymphoma in which the majority of patients are cured with conventional chemotherapy (NCCN Guidelines NHL2014).

[0175] First-line therapy for DLBCL typically has an objective response rate of approximately 80% and a complete response rate of approximately 50%. Anthracism using rituximabs such as R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) has a % (Coiffier 2002). Including the Clin-containing method, approximately one-third of patients have an anti-treatment disease or relapse after R-CHOP (Sehn 2005). For patients who relapse after responding to first-line therapy, Approximately 40-60% of patients can achieve a second response with additional chemotherapy. Standard treatment for second-line therapy for patients eligible for autologous stem cell transplantation (ASCT) includes rituximab and combination chemotherapy such as R-ICE (rituximab, ifosfamide, carboplatin, and etoposide) and R-DHAP (rituximab, dexamethasone, cytarabine, and cisplatin), which have objective response rates of approximately 63% and complete response rates of approximately 26% (Gisselbrecht). Patients who respond to second-line therapy and those deemed well-suited for transplantation receive consolidation with high-dose chemotherapy and ASCT, which cures about half of transplanted patients (Gisselbrecht 2010). Patients who fail ASCT have a very poor prognosis and are not considered a curative choice. It has no limbs.

[0176] Primary mediastinal large B-cell lymphoma (PMBCL) has distinct clinical and pathological characteristics compared to DLBCL. It has molecular characteristics. PMBCL is thought to originate from thymic (medulla) B cells and accounts for approximately 3% of patients diagnosed with DLBCL. PMBCL is typically seen in relatively young adults in their 40s. It has been observed in the population, with a slightly higher prevalence in women. Gene expression profiling suggests that PMBCL is associated with The deregulated pathways in PMBCL are suggested to overlap with those in Hodgkin lymphoma. Initial therapy generally includes vincristine, prednisone, and rituximab-containing anthracycline formulations such as vincristine, prednisone, and rituximab-containing anthracycline formulations such as retuximab-containing anthracycline formulations such as etoposide, doxorubicin, and cyclophosphamide (DA-EPOCH-R), with or without regional irradiation therapy.

[0177] Follicular lymphoma (FL), a type of B-cell lymphoma, is the most common indolent (slowly progressing) form of NHL, accounting for approximately 20-30% of all NHL cases. Some patients with FL have more Aggressive, histologically transforms into DLBCL (TFL), which is associated with poor outcomes. Histological transformation occurs at an annual rate of approximately 3% over 15 years, and the risk of transformation is high in the following years. It continues to decline. The biological mechanism of histological transformation is unknown. Initial treatment for TFL is While influenced by prior treatment for follicular lymphoma, treatment generally involves anthracycline-containing formulations in combination with rituximab to eliminate the aggressive component of this disease.

[0178] Treatment options for relapsing / antitherapy-treated PMBCL and TFL are similar to those for DLBCL. Given the low prevalence of these diseases, large-scale prospects in these patient populations are expected. No randomized trials have been conducted. Patients with chemotherapy-treated diseases are treated with anti-treatment DLBCL. Patients with this condition have a similar or worse prognosis.

[0179] In summary, individuals with anti-treatment aggressive NHL (e.g., DLBCL, PMBCL, and TFL) have a significant unaddressed medical need, and further research using novel treatments in these populations is warranted.

[0180] Therefore, in some embodiments, this method can be used to treat lymphoma or leukemia, in which case lymphoma or leukemia is a B-cell malignancy. In this context, lymphoma or leukemia includes B-cell chronic lymphocytic leukemia / small cell lymphoma, B-cell prelymphocytic leukemia, lymphoplasmacytic lymphoma (e.g., Waldenström macroglobulinemia), splenic marginal lamina lymphoma, hairy cell leukemia, plasmacytic neoplasms (e.g., plasmacytic myeloma (i.e., multiple myeloma), or plasmacytoma), extranodal marginal zone B-cell lymphoma (e.g., MALT lymphoma), nodal marginal zone B-cell lymphoma, follicular lymphoma (FL), morph Transformed follicular lymphoma (TFL), primary cutaneous follicular lymphoma, mantle cell lymphoma Diffuse large B-cell lymphoma (DLBCL), Epstein-Barr virus-positive DLBCL, lymphomatous granulomatosis, mediastinal (thymic) large B-cell lymphoma (PMBCL), intravascular large B-cell lymphoma Cellular lymphoma, ALK+ large B-cell lymphoma, plasmablastic lymphoma, primary exudative lymphoma Lymphoma, large B-cell lymphoma occurring in HHV8-associated multicentric Castleman disease, Kitt lymphoma / leukemia, T-cell pre-lymphocytic leukemia, T-cell macrogranular lymphocytic leukemia, A Aggressive NK cell leukemia, adult T cell leukemia / lymphoma, extranodal NK / T cell lymphoma, enteropathy-associated T cell lymphoma, hepatosplenic T cell lymphoma, blastic NK cell lymphoma, mycosis fungoides / Sézary's disease Symptoms, primary cutaneous anaplastic large cell lymphoma, lymphomatoid papulosis, peripheral T-cell lymphoma, hematological Idioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, B-cell lymphoblastic leukemia / lymph B-cell lymphoblastic leukemia / lymphoma with recurrent genetic abnormalities, T-cell lymphoblastic leukemia The choice is made from hematological / lymphoma and Hodgkin lymphoma. In some aspects, cancer is , resistant to one or more previous treatments, and / or cancer, one or more This is a recurrence following previous treatment.

[0181] In certain embodiments, the cancer is selected from follicular lymphoma, transformed follicular lymphoma, diffuse large B-cell lymphoma, and mediastinal (thymic) large B-cell lymphoma. In a particular embodiment, the cancer is diffuse large B-cell lymphoma.

[0182] In some aspects, cancer is treated with chemotherapy, radiotherapy, immunotherapy (T-cell therapy and / or other treatments). Treatments including those using antibodies or antibody-drug conjugates, autologous stem cell transplantation, or any combination thereof, one or more of which are anti-therapeutic or the cancer subsequently It is a recurrence. In a specific form, the cancer is an anti-treatment diffuse large B-cell phosphate cancer. It is paprano cancer.

[0183] The present invention is further illustrated by the following embodiments, which should not be construed as limiting. The contents of all references cited herein are expressly incorporated herein by reference. [Examples]

[0184] Example 1 A phase 1 / 2 single-arm, open-label trial was planned to determine the safety and feasibility of administering anti-CD19CAR+ T cells to subjects with B-cell malignancies.

[0185] Subjects who sign informed consent and meet the trial entry criteria will be enrolled in the trial and will undergo leukocyte apheresis to obtain PBMCs for anti-CD19 CAR+ T cell production. Prior to hospitalization for a single infusion of anti-CD19 CAR+ T cells on day 0, subjects were treated with conditioning chemotherapy. Next, three hours after the infusion of anti-CD19 CAR+ T cells, some subjects were treated with interleukin-2 (group 1 only). If a partial response (PR) or complete response (CR) was observed after the first infusion, and disease progression continued thereafter, retreatment with a second dose of anti-CD19 CAR+ T cells was permitted.

[0186] Three groups of subjects were enrolled. Group 1 consisted of 3 × 10⁶ anti-CD19 CAR+ T cells. 6 ~30×10 6 This study included eight subjects who received anti-CD19 CAR+ T cells in the range of cells / kg, including one subject who underwent retreatment. The dose of anti-CD19 CAR+ T cells ranged from 60 to 120 mg / kg (2220 to 4440 mg / m²). 2 ) High-dose cyclophosphamide for 2 days In between, followed by 25 mg / m² 2 Fludarabine was administered to subjects following a 5-day conditioning regimen. These subjects received 720,000 IU / kg of anti-CD19 CAR+ T cells to stimulate their proliferation after administration. They also received high-dose interleukin-2 (IL-2) (up to 15 doses, or every 8 hours until toxicity prevented further doses).

[0187] Group 2 received high doses of cyclophosphamide and fludarabine, and various doses Administration of anti-CD19 CAR+ T cells (1 x 10⁶ anti-CD19 CAR+ T cells) 6 ~5×10 6 (pieces / kg) after interloying This group includes 15 subjects who did not receive N-2, including 2 subjects from group 1 who were retreated.

[0188] Group 3 is a reduced conditioning regimen of 300 mg / m². 2 Cyclophosphamide 30 mg / m² 2 This includes 11 subjects who received fludarabine (all administered concurrently for 3 days without IL-2 administration). Of these subjects, the first 7 were 1 × 10⁶ 6 Individual anti-CD19 CAR+ T cells, later 4 People are 2 x 10 6 The patient received an injection of anti-CD19 CAR+ T cells.

[0189] demographics Table 1 provides demographic and disease characteristics of the subjects. Of the 32 subjects enrolled, 19 (59%) had DLBCL or PMBCL, 7 (22%) had CLL, and 6 (19%) had other indolent NHL, including indolent follicular lymphoma and splenic marginal lymphoma. The subjects in this group had a medically treatable disease (84%) and had previously received a median of three series of treatments. All subjects with sibiric NHL had previously received anti-CD20 therapy and platinum-based chemotherapy, and 95% She had previously received anthracycline-based chemotherapy.

[0190] Pharmacokinetics Using qPCR analysis, the number of anti-CD19 CAR+ T cells in peripheral blood at various time points after the initial administration on day 0 was evaluated and validated by standard curves created by flow cytometry using an antibody reagent specific to scFv present in the anti-CD19 CAR construct (Kochenderfer et al., "B-cell depletion and remissions of malignancy along with cytokine-associated toxicity in a clinical trial of anti-CD19 chimeric-antigen-receptor-transduced T cells," Blood 119:2709-20 (2012)).

[0191] (Table 1) Demographics of the subjects of the clinical trial TIFF0007867521000001.tif146166

[0192] In group 1, anti-CD19 CAR+ T cells 3 × 10 6 ~30×10 6 Cells / kg were injected. In the first six subjects, anti-CD19 CAR+ T cells in the circulating blood reached a maximum of 0.02-1% of the total PBMCs within two weeks after injection. They were detected at higher levels, then rapidly disappeared, and were undetectable after 50 days. The largest number of anti-CD19 CAR+ T cells (28 × 10⁶ anti-CD19 CAR+ T cells each) 6 cells / kg and anti-CD19 CAR+ T cells 30×10 6 Subjects 7 and 8, given (pieces / kg), reached >10% of the total PBMC, They possessed a high peak percentage of anti-CD19 CAR+ T cells and had longer survival times of anti-CD19 CAR+ T cells in the blood (>130 days and 180 days, respectively).

[0193] In group 2, in the absence of interleukin-2 treatment, anti-CD19 CAR+ T cells were reduced for 2 weeks. The cells showed similar proliferation in the peripheral blood, followed by disappearance, and were completely eliminated from circulation within a few weeks (Table 2).

[0194] Overall, the relationship between the dose of anti-CD19 CAR+ T cells and their proliferation and survival in peripheral blood There was no clear relationship between them. Similarly, so far, anti-CD19 CAR+ T cells The dose, the proliferation or survival duration of anti-CD19 CAR+ T cells in the blood, and the effects of this treatment. There was no clear relationship between clinical response or toxicity.

[0195] (Table 2) Proliferation and survival duration of anti-CD19 CAR+ T cells in peripheral blood of subjects in Group 2 TIFF0007867521000002.tif31156

[0196] In groups 1 and 2, there was no secondary proliferation of anti-CD19 CAR+ T cells after primary proliferation 7–14 days after injection. In previously tested subjects, genome insertion of CAR-expressing retroviruses occurred. There is no evidence of carcinogenic transformation. The results for group 3 are still not relevant at the time of the data cutoff. I hadn't been able to get it.

[0197] efficacy Clinicians evaluated safety in 32 subjects and efficacy in 29 subjects. The overall response rate for the 29 subjects for whom efficacy could be evaluated was 76%. Of the 29 subjects, 11 (38%) achieved complete response (CR), and 11 out of 29 subjects (38%) achieved partial response (PR) (Figures 2A and 2B; Table 3).

[0198] Of the 29 evaluable subjects, 16 (55%) remained in response from their initial treatment, and the duration of response for 12 subjects (including those who received retreatment) exceeded one year (Table 3). Of the 3 subjects who responded, These patients were re-treated after disease progression, and all are still showing a positive response (over 17.4 to 52.2 months).

[0199] As shown in Table 3, of the 19 subjects with antitherapy-responsive aggressive DLBCL / PMBCL, 17 were evaluable for disease response (1 subject was not evaluable; 1 subject had not yet been evaluated). Of these 17 subjects, 11 (65%) showed a response, and 6 out of 17 (35%) achieved a complete response (CR). This was achieved. The median duration of response was 7.3 months.

[0200] (Table 3) Objective response rate and duration of response for each tumor type The "+" in TIFF0007867521000003.tif88161 indicates that the effect is still ongoing.

[0201] Of the seven evaluable subjects with CLL, six (86%) showed a response, and four out of seven (57%) achieved a complete response (CR) (Table 3). The median duration of response was 22.2 months, with responses lasting longer than 27 months. Four out of seven subjects (57%), including the three subjects mentioned above, are still experiencing a positive response (Table 3).

[0202] Of the five evaluable subjects with indolent NHL, all five (100%) showed a response, and one subject 20% of patients achieved a complete response (CR). The median duration of response was 18.8 months (Table 3). Five patients (5 out of 5; 100%) continued to respond, and two patients maintained a response for longer than 45 months (Table 4).

[0203] safety Adverse events The 32 subjects were treated with anti-CD19 CAR+ T cells, and the last subject treated experienced no adverse events. No reports have been made yet. The overall safety summary includes all 32 subjects treated. The group-specific summaries are for subjects 1010003 and 1010004, twice (the first time when these subjects were treated in group 1 and the second time when these subjects were treated in group 2) (using anti-CD19 CAR+ T cells). Includes safety data for retreatment.

[0204] Summary of Adverse Events Table 4 provides an overview of adverse events. Overall, 31 subjects (97%) experienced one or more adverse events. Of those who experienced the elephant, 0 people (0%) experienced Grade 3, the worst grade, and 29 people (91%) experienced Grade 3, the worst grade. A grade 4 adverse event was experienced in the worst-case scenario, with 2 subjects (6%) experiencing a fatal adverse event. 20 subjects (63%) experienced adverse events related to anti-CD19 CAR+ T cells; 6 subjects (19%) experienced the worst-case scenario as grade 3, 8 subjects (25%) experienced the worst-case scenario as grade 4, and no subjects experienced a grade 5 event. 16 subjects (50%) experienced serious adverse events; 3 subjects (9%) experienced the worst-case scenario as grade 3, and 9 subjects (28%) experienced the worst-case scenario. The average was Grade 4, with 2 people (6%) experiencing the worst-case scenario of Grade 5.

[0205] Dose-limiting toxicity The incidence of DLTs in groups 1, 2, and 3 was 38%, 40%, and 0%, respectively. Except for subject 1010002, DLTs were primarily neurotoxic, with two cases of elevated creatinine and one each of hypoxia and hypotension. Table 6 provides a list of DLTs. In group 3, DLTs were reported. There wasn't one. In group 3, 2 × 10 6 Conditioning method using anti-CD19 CAR+ T cells / kg We tested it.

[0206] (Table 4) Summary of adverse events TIFF0007867521000004.tif90155

[0207] Cytokine release syndrome Cytokine release is induced by activated T cells associated with CD19 targets. Using the search strategy, adverse events that may be attributable to CRS that occurred after treatment include fever and febrile neutrophils. Adverse events included cytopenia, hypotension, acute vascular leak syndrome, elevated creatinine, renal failure, hypoxia, and pleural effusion. Adverse events, possibly attributable to cytokine release, were reported in 28 subjects (88%), with ≥grade 3 events reported in 24 subjects (75%), and 6 subjects (19%) experiencing serious events. IL-2 (used in group 1) and conditioning chemotherapy. Adverse events from combination therapies, such as those causing febrile neutropenia, may potentially confound this analysis.

[0208] Clinical signs of CRS typically occur in the first week after anti-CD19 CAR+ T cell infusion, and in group 3 pairs It was not commonly observed in elephants. Only one out of 11 subjects in group 3 experienced grade 3 hypotension. Four individuals experienced grade 3 fever. Acute vasoleak syndrome, oliguria, elevated creatinine, and renal failure were reported only in subjects in groups 1 and 2.

[0209] (Table 5) Dose-limiting toxicity TIFF0007867521000005.tif111155

[0210] Neurological adverse events Neurological adverse events, primarily aphasia / language disorders, confusion, motor neuropathy, and somnolence, were observed in all three groups. Thirteen subjects (41%) experienced severe neurotoxicity of ≥grade 3. Eleven of the subjects (34%) experienced serious events.

[0211] The subjects who died exhibiting neurotoxicity had experienced CNS cerebral vascular ischemia in the context of influenza A virus infection. The principal investigator considered this unrelated to anti-CD19 CAR+ T cells. did.

[0212] Five subjects (16%) who experienced neurotoxic events required airway management for neurological adverse events. They required mechanical ventilation, and all of these subjects were in groups 1 and 2. Group 3 was intubated. There were no suitable subjects.

[0213] Neurological adverse events occurred between day 2 and day 17 after anti-CD19 CAR+ T cell infusion, with a median duration of 6 days. It occurred in the eye. However, it occurred in one subject, 110 days after anti-CD19 CAR+ T cell injection. Excluding Grade 4 myelitis. Based on the time of onset, symptoms, and brain MRI findings, the principal investigator concluded that this event was related to fludarabine and not caused by anti-CD19 CAR+ T cells. The median time for neurological adverse events to resolve to grade 1 or lower was 14 days after infusion. That was the case.

[0214] death Two subjects died within 30 days of receiving chemotherapy and anti-CD19 CAR+ T-cell infusion. However, 18 days after the treatment during the clinical trial, the patient died from viral pneumonia, influenza A infection, E. coli infection, respiratory distress, and a stroke that occurred simultaneously with hypoxia. Subject 11 had extensive symptoms. The patient had PMBCL with fibrous mediastinal lymphoma spread and died 16 days after the treatment during the clinical trial. Autopsy The cause of death was not determined at the time, and the autopsy report stated that, considering the mediastinal spread of PMBCL, the possible cause of death was The cause was concluded to be cardiac arrhythmia. The principal investigator concluded that all events were related to anti-CD19 CAR+ T cells. Then they didn't see it.

[0215] Example 2 Selected patients will receive cyclophosphamide 300 mg / m². 2 / day and fludarabine 30 mg / m² 2 / day Conditioning chemotherapy was administered. Conditioning chemotherapy was given for 3 days from day -5 to day -3. On day 0, the first subset of patients (patients 22-28) (Table 6) received new medication manufactured for 10 days. After receiving administration of anti-CD19 CAR+ T cells, a second subset of patients (patients 29-32) produced 6 days of production. The patient received administration of cryopreserved anti-CD19 CAR+ T cells.

[0216] Patient serum was examined using Luminex with the Millipore HCD8MAG15K17PMX kit (T1, T2, immunomodulatory cytokines, chemokines, immunoeffectors). Interleukin-15 (IL-15) and monocyte chemotactic protein 1 (MCP-1) were administered before and after the procedure. The levels of gamma-inducible protein 10 (IP-10), placental growth factor (PLGF), soluble intercellular adhesion molecule 1 (sICAM-1), C-reactive protein (CRP), vascular endothelial growth factor D (VEGF-D), and macrophage inflammatory protein 1β (MIP-1β) were measured.

[0217] (Table 6) Data on the condition and outcomes of patients 22-28 TIFF0007867521000006.tif61157DLBCL = diffuse large B-cell lymphoma; FL = follicular lymphoma; PR = partial response; CR = complete response; PD = progressive disease

[0218] Of patients 22-28, patients 22-25 and 27 showed at least a partial response, while patients 26 and 28 developed progressive disease after treatment. For patients 22-26, serum levels of IL-15 and MCP-1 Levels of IL-15 and PLGF showed at least some increase (Figures 4A, 4B, and 4D), while levels of IP-10, sICAM-1, CRP, VEGF-D, and MIP-1β increased in some patients and remained stable or decreased in others (Figures 4C and 4E-4H). Only IL-15 was measured for patients 27 and 28 (Figure 4A).

[0219] Several differences in marker levels were observed between responding patients showing either partial or complete response and non-responding patients with progressive disease. IL-15 levels were baseline. In contrast, in responding patients, the IL-15 level increased by an average of approximately 35 times, ranging from approximately 10 times to approximately 55 times, while in non-responding patients, the increase in IL-15 levels was less than approximately 10 times (Figure 5A). In patients with a response, MCP-1 levels increased by an average of approximately 5 times, ranging from approximately 2 times to approximately 7 times, while in patients without a response (Patient 26), the increase in MCP-1 levels was less than 4 times (Figure 5B). IP-10 in patients with a response The level increased by an average of about 3.5 times, ranging from about 2 times to about 7 times, while those who did not respond... There was essentially no change in serum IP-10 levels (Figure 5C). PLGF levels in responders increased by an average of about 30 times, ranging from a slight increase of less than 2 times to an increase of more than 100 times, while non-responders showed only a slight increase in serum PLGF levels (Figure 5D). sICAM-1 levels in responders increased by an average of about 3 times, ranging from essentially unchanged to an increase of about 4.5 times. In contrast, serum sICAM-1 levels remained essentially unchanged in non-responders (Figure 5E). CRP levels in responders increased by an average of approximately 10 times, ranging from essentially unchanged to an increase of approximately 25 times, while serum CRP levels remained essentially unchanged in non-responders (Figure 5F). VEGF-D levels in responders increased by an average of approximately 3 times, ranging from essentially unchanged to an increase of approximately 6 times, while In non-responders, serum VEGF-D levels were essentially unchanged (Figure 5G). In responders, MIP-1β levels increased by an average of approximately 1.5 times, ranging from essentially unchanged to an increase of approximately 3 times. On the other hand, serum MIP-1β levels decreased by only about 50% in non-responders (Figure 5H).

[0220] Patients 30-33 were given cryopreserved cells prepared over 6 days, and selected days from day -6 to day 18. In addition, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFNγ or IFNG), interleukin-10 (IL-10), IL-15, interleukin-2 (IL-2), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-8 (IL-8), IP-10, MCP-1, MIP-1β, serum granzyme A (GRNZA), serum granzyme B (GRNZB), PLGF, CRP, monocyte chemotactic Levels of various cytokines, chemokines, effectors, inflammatory markers, and adhesion molecules were measured, including protein 4 (MCP-4), interleukin 16 (IL-16), thymic and activation-regulated chemokines (TARC), eotaxin-3, sICAM-1, soluble vascular adhesion molecule 1 (sVCAM-1), and serum amyloid A (SAA) (Figures 6A-6V).

[0221] Example 3 To improve the degree and duration of lymphocyte depletion observed in Group 3 of Example 1, the dose of conditioning chemotherapy in Cohort A1 was changed to cyclophosphamide 500 mg / m². 2 and fludarabine 30 mg / m² 2 The target dose for anti-CD19 CAR+ T cells is increased to 2 × 10⁶. 6 The drugs were administered simultaneously at a dose of 1 / kg for 3 days. The dose of cyclophosphamide used in this method (Cohort A1) was approximately 38% lower than the dose used in the conditioning method of cyclophosphamide 30 mg / kg in Group 2 from Example 1 (incidence of dose-limiting toxicity (DLT) 29%), and was lower than the same dose of fludarabine used in Group 3 of Example 1.

[0222] The evaluation of higher doses of conditioning chemotherapy and / or various doses of anti-CD19 CAR+ T cells will proceed based on the assessment of DLT incidence and benefit. The CAR vector construct is identical to the construct described in Example 1. This example uses a rapid, closed-loop, bead-free process. Therefore, a clinical trial was designed to test the safety and efficacy of the generated anti-CD19 CAR+ T cells. The experiment is described below. Upon completion of the process, the characteristics of the T cell product are preserved.

[0223] Experimental Design A multicenter Phase 1 / 2 study to evaluate the safety and efficacy of KTE-C19 in patients with anti-treatment NHL. An open-label trial will be conducted. This trial will be divided into two distinct phases, referred to as Phase 1 and Phase 2. ru.

[0224] During Phase 1, approximately 6 to 24 patients with DLBCL, PMBCL, or TFL will be enrolled to evaluate the safety of the KTE-C19 method. The sponsor's internal Safety Review Team (SRT) will review the safety data. The study conducted a review and, as shown in Figure 3, recommended further implementation of the Phase 1 trial and progression to Phase 2. cormorant.

[0225] During Phase 2, subjects will be enrolled in two separate cohorts, referred to as Cohort 1 and Cohort 2. Adult subjects with anti-treatment DLBCL will be enrolled in Cohort 1, and adult subjects with anti-treatment PMBCL and TFL will be enrolled in Cohort 2. TFL is defined as subjects who have previously received chemotherapy for follicular lymphoma.

[0226] Regardless of the phase of the study, each subject will follow the same study treatment schedule and procedural requirements. Each subject will undergo the following study periods: screening / leukocyte apheresis period; conditioning chemotherapy period; investigational drug (IP) treatment period; post-treatment evaluation period; and long-term follow-up period. To move forward by going through various experiences.

[0227] Test duration For individual subjects, the length of participation is as follows: a maximum screening period of 28 days, a conditioning chemotherapy treatment period of 5-7 days, and a KTE-C19 treatment period (including a 7-day hospitalization and recovery period). This includes the post-treatment assessment period and the long-term follow-up period (a maximum of 15 years of survival surveillance). nothing.

[0228] The subjects will be followed up for 3 months after treatment for all adverse events. After 3 months, the subjects will be followed up at intervals outlined in the evaluation schedule (SOA) for targeted adverse events / serious adverse events (e.g., hematological, neurological, secondary malignancies, infections, or autoimmune disorders) and the presence of replicable retroviruses (RCRs) in the subjects' blood. The need for long-term follow-up will be determined based on the treatment. This is based on the potential persistence of the gene transfer vector in the target organism.

[0229] Completion of the trial is defined as the time when the last subject completes their long-term follow-up visits, is deemed to have failed follow-up, withdraws consent, or dies. Regardless of which occurs first, all subjects in Phase 2 Cohort 1 and the overall trial population will have completed the 6-month disease response assessment, failed follow-up, discontinued the trial, or died. A primary analysis is performed upon death.

[0230] Eligibility The inclusion criteria for the subjects include the following: a) Histologically confirmed aggressive B, including the following types as defined by WHO 2008: Cellular NHL: DLBCL nonspecific type, T cell / histiocyte-rich B large cell lymphoma, DLBCL associated with chronic inflammation, Senile Epstein-Barr virus (EBV) + DLBCL; mediastinal (thymic) large cell B lymphoma; or transformation of follicular lymphoma into DLBCL; b) Stable disease state as the best response to the most recent chemotherapy regimen (duration of stable disease state) (must be ≤12 months) or progressive disease; and ≤12 months from previous self-SCT A chemotherapy-treated disease defined as one or more of the following: disease progression or relapse; c) Participants must have received sufficient prior treatment, including anti-CD20 monoclonal antibody and anthracycline-containing chemotherapy, unless the principal investigator determines that the tumor is CD20-negative; d) Subjects with transformed FL have previously received chemotherapy for follicular lymphoma, Furthermore, the patient must have an anti-chemotherapy-responsive disease after transformation to DLBCL; e) At least one measurable lesion according to the revised IWG criteria for evaluating the effectiveness of treatment for malignant lymphoma; previously irradiated lesions are measured only if progression is demonstrated after completion of radiotherapy. To consider possible; f) Brain MRI that does not show evidence of central nervous system lymphoma; g) At the time leukocyte apheresis is planned for the subject, at least two weeks must have passed since any previous radiotherapy or systemic therapy; h) Toxicity from previous treatments must be stable or have recovered to ≤ Grade 1 (excluding clinically insignificant toxicity such as alopecia); i) The target group must be 18 years of age or older; j) Performance status of the Eastern Cooperative Oncology Group (ECOG) is 0 or 1; k) Subjects must have the following clinical laboratory values: (i) ANC ≥ 1000 / uL; (ii) Platelet count ≥ 50,000 / uL; (iii) Serum creatinine ≤ 1.5 mg / dL, serum ALT / AST ≤ 2.5 ULN, and Gilbert's Sufficient renal, hepatic, and cardiac function, defined as total bilirubin ≤ 1.5 mg / dl, excluding subjects with the syndrome; and iv) cardiac ejection fraction ≥ 50% and no evidence of pericardial effusion, as determined by echocardiography; and l) Women of childbearing potential must have a negative result on a serum or urine pregnancy test.

[0231] The exclusion criteria for those subject to exclusion include the following: a) A history of malignant disease other than non-melanoma skin cancer or carcinoma in situ (e.g., cervical, bladder, or breast cancer) or follicular lymphoma, unless the patient has been disease-free for at least three years; b) A history of Richter transformation in CLL; c) Autologous stem cell transplantation within 6 weeks of informed consent; d) History of allogeneic stem cell transplantation; e) Except for subjects who received KTE-C19 in this study and are eligible for retreatment, previously targeted CD19 Therapy; f) Previous chimeric antigen receptor therapy or other genetically modified T-cell therapy; g) A history of severe immediate-type hypersensitivity reactions caused by aminoglycoside drugs; h) Clinically significant active infections (e.g., simple UTIs, bacterial pharyngitis are acceptable) You are currently receiving intravenous antibiotics or have received intravenous antibiotics within 7 days prior to registration. (Prophylactic antibiotics, antivirals, and antifungals are permitted); i) A history of infection with HIV, hepatitis B virus (HBsAg positive), or hepatitis C virus (anti-HCV positive) I understand; j) Subjects who have detectable malignant cells in the cerebrospinal fluid or brain metastases, or who have a history of malignant cells in the cerebrospinal fluid or brain metastases; k) A history of paroxysmal disorder, cerebrovascular ischemia / hemorrhage, dementia, cerebellar disorder, or any autoimmune disease affecting the CNS; l) Patients whose lymphoma has spread to the atria or ventricles; m) Emergency treatment is required due to a mass effect such as intestinal obstruction or vascular compression; n) Primary immunodeficiency; o) Any medical condition that may interfere with the determination of the safety or efficacy of the test procedure; p) Systemic corticosteroid therapy currently required or expected to be required; for patients with adrenal insufficiency Standard doses of topical and inhaled corticosteroids for elephants, as well as physiological substitutes, are acceptable; prednisone doses of 5 mg / day or more of other corticosteroids An equivalent dose is not permitted; q) A history of severe immediate-type hypersensitivity reactions to any of the drugs used in this study; r) Live vaccine administered ≤ 6 weeks before the start of the conditioning regimen; s) Pregnant or breastfeeding women of childbearing potential due to the potential adverse effects of preliminary chemotherapy on the fetus or infant; women who have been sterilized or who are at least two years postmenopausal and therefore of childbearing potential. Not considered; t) Subjects of both sexes who do not wish to undergo contraception from the time of agreement until 6 months after the completion of KTE-C19; and u) The principal investigator determines that the subject is unlikely to complete any of the study examinations or procedures required for the protocol, including follow-up examinations, or is unlikely to comply with the participation requirements for the study.

[0232] In addition, biomarker analysis was performed on blood and tumor samples to determine the status of KTE-C19. Evaluate predictive and pharmacodynamic markers. Prognostic markers in aggressive NHL. —The following may also be evaluated: baseline leukocyte apheresis sample and final KTE-C19 The sample is deposited and analyzed by immunophenotyping and / or gene expression profiling. The remaining sample may be saved for future exploratory analysis of DNA markers, RNA markers, or protein markers. The saved tumor tissue is collected for central path review. Additional analyses may include CD19 expression and gene expression profiling. This may include analysis of DNA changes, DNA markers, RNA markers, or proteins. The remaining tumor sample may be saved for future exploration and analysis of markers.

[0233] Treatment protocol schedule Leukocyte apheresis (for the production of KTE-C19) using approximately 5 x 10 mononuclear cells 9 ~10×10 9 Targeting individuals The goal is to obtain white blood cells from the subjects by apheresis of 12-15 liters. The leukocyte apheresis product is processed to concentrate the T cell-containing PBMC fraction. Next, the T cells are stimulated to proliferate, and a retroviral vector is transduced into them to introduce the CAR gene. Next, T cells are proliferated, cryopreserved, and used to generate the test product. Each target's condition After the completion of the conditioning chemotherapy regimen, each patient receives their respective KTE-C19 injection.

[0234] Test treatment The subjects received a non-myeloablative conditioning regimen consisting of cyclophosphamide and fludarabine to induce lymphocyte depletion, creating an optimal environment for KTE-C19 proliferation. This will be done in vivo. The subjects will start on day -5 (or day -7 for cohort B) and then on day -1 Conditioning chemotherapy using cyclophosphamide and fludarabine will be initiated over the course of a day. The 5-day conditioning chemotherapy regimen will be administered on an outpatient basis. The 7-day conditioning chemotherapy regimen may be administered to outpatients or inpatients at the discretion of the principal investigator. It can be administered as an inpatient treatment.

[0235] Phase 1: In cohorts A1 and A2, the subjects underwent the following 5-day conditioning chemotherapy regimen. Receive the prescribed administration: On the day of infusion, before administering cyclophosphamide, take 1 liter of 0.9% NaCl saline solution. Hydration using IV; followed by 500 mg / m² on days -5, -4, and -3. 2 Cyclophosph Administer amide intravenously over 60 minutes; followed by 30 mg / m² on days -5, -4, and -3. 2 Fludarabine is administered intravenously over 30 minutes; followed by an additional 1 liter of 0.9% NaCl saline upon completion of the fludarabine infusion (Figure 3). In certain cases, according to facility guidelines, mesna (2-mercaptoethanesulfone) may be used. Sodium phosphate can be added.

[0236] In cohort A3, subjects received the following 5-day chemotherapy regimen: intravenous fluid replacement with 1 liter of 0.9% NaCl saline administered prior to cyclophosphamide on the infusion day; followed by 300 mg / m² on days -5, -4, and -3. 2 Administer cyclophosphamide IV over 60 minutes; followed by 30 mg / m² on days -5, -4, and -3. 2 Fludarabine is administered intravenously over 30 minutes; followed by an additional 1 liter of 0.9% NaCl saline upon completion of the fludarabine infusion. In certain cases, mesna may be added according to facility guidelines.

[0237] For subjects registered in cohorts A1, A2, or A3, day -2 and day -1 are treated as day 0. This is a rest day before the KTE-C19 injection.

[0238] In cohorts B1 and B2, subjects received the following 7-day chemotherapy regimen: intravenous fluid replacement with 0.9% NaCl saline at a recommended rate of 2.6 ml / kg / hr (maximum 200 ml / hr) as a continuous infusion starting 11 hours before cyclophosphamide infusion, and continuing fluid replacement until 24 hours after the final cyclophosphamide infusion; 30 mg / kg (1110 mg / m³) on days -7 and -6. 2 Administer cyclophosphamide intravenously over 120 minutes; followed by 25 mg / m² on days -5, -4, -3, -2 and -1. 2 Fludarabine is administered intravenously, infused over 30 minutes. In certain cases, mesna may be added according to facility guidelines.

[0239] For subjects enrolled in cohort B1 or B2, there is no rest day between the final day of chemotherapy (day -1) and the KTE-C19 infusion on day 0.

[0240] For KTE-C19, the subjects in cohorts A1, A3, or B1 received a target dose of 2 × 10⁶ anti-CD19 CAR+ T cells. 6 cells / kg (±20%; anti-CD19 CAR+ T cells 1.6×10 6 cells / kg~anti-CD19 CAR+ T cells 2.4×10 6 The KTE-C19 treatment consists of a single infusion of CAR-transduced autologous T cells administered intravenously at a dose of (cells / kg). The minimum dose of anti-CD19 CAR+ T cells is 1 × 10⁶. 6 It may be administered at doses per kg. If body weight exceeds 100 kg For the target group, the maximum uniform dose of anti-CD19 CAR+ T cells is 2 × 10⁶. 8 Individual doses are administered.

[0241] In cohort A2 or B2, the subjects received a targeted dose of 1 × 10⁶ anti-CD19 CAR+ T cells. 6 pieces / kg(±20 %;anti-CD19 CAR+ T cells 0.8×10 6 cells / kg~anti-CD19 CAR+ T cells 1.2×10 6 The patient receives KTE-C19 treatment, which consists of a single infusion of CAR-transduced autologous T cells administered intravenously at a dose of 0.5 × 10⁶ cells / kg. The patient also receives the minimum dose of anti-CD19 CAR+ T cells, which is 0.5 × 10⁶. 6 The dose may be cells / kg. For subjects weighing over 100 kg, the maximum uniform dose of anti-CD19 CAR+ T cells is 1 × 10⁶. 8 Individual doses are administered.

[0242] Phase 2: The KTE-C19 method, which was determined to be safe in Phase 1 by SRT, will be advanced to Phase 2 of the test.

[0243] Retreatment Patients who achieve a partial response (PR) or complete response (CR) will receive a second course of conditioning chemotherapy and KTE-C19 if the disease subsequently progresses (and the relapse is not known to be due to CD19-malignant cells). They can receive it. In order to be eligible for the second course of treatment, the subject will be re-evaluated and Except for the previous exclusion criteria for CAR therapy, it should continue to meet the original eligibility criteria for the study. Furthermore, the patient should not have received subsequent chemotherapy for the treatment of lymphoma. In addition, any toxicity related to fludarabine or cyclophosphamide, except for alopecia, should be stable or have recovered to less than grade 1 before retreatment. Maximum of one per patient. A retreatment cycle may occur. Subjects enrolled in Phase 2 will receive the same KTE-C19 regimen. Subjects enrolled in Phase 1 will receive the KTE-C19 regimen selected for Phase 2. If a Phase 2 regimen has not yet been selected, participants will receive the final KTE-C19 regimen deemed safe by SRT.

[0244] Subjects who experienced DLT in Phase 1, or comparable toxicity in Phase 2, The subject is ineligible for retreatment. Furthermore, if the subject is known to have neutralizing antibodies, the subject is ineligible for retreatment. However, if non-neutralizing HAMA or HABA antibodies are present, the subject may be retreated if it meets the eligibility criteria.

[0245] Post-treatment assessment After completing KTE-C19 infusion and being discharged (typically on day 8), all subjects are monitored during the post-procedure evaluation period. Counting from day 0 (KTE-C19 infusion), subjects are monitored at week 2 and week 4 (±3 days). You will return to the clinic in the second month (±1 week) and the third month (±1 week). The results will be determined by the MMSE (Mini-Mental State Examination). Tal state examination; PET-CT for disease diagnosis; physical examination and vital signs; clinical laboratory tests including chemical panel, CBC blood smear with differential diagnosis, β-HCG pregnancy test (serum or urine) for all women of childbearing potential, anti-KTE-C19 antibody, lymphocyte subset, cytokine levels, anti-CD19 CAR+ T cells, and replicable retrovirus (RCR) analysis; reporting of adverse / serious adverse events; concomitant medications Description; and including the collection of fresh tumor samples from subjects who have signed any part of the agreement. It is possible.

[0246] Abundance, proliferation, survival, and immunosuppression of transduced anti-CD19 CAR+ T cells in the blood The actual organism is primarily detected and monitored by PCR analysis, and then captured by flow cytometry. The levels of serum cytokines will also be assessed from the blood. The following cytokines may be included in the panel: pro-inflammatory and immunomodulatory cytokines IL-6, TNFα, IL-8, IL-1, IL-2, GM-CSF, IL-15, IL-17a, IFNγ, IL-12p40 / p70; immunoeffector molecules granzyme A, B, perforin, sFasL; correlations of the acute phase response CRP, SAA, and chemokines MIP-1α, MIP-3α, IP-10, eotaxin, MCP-4. KTE-C19 is a retroviral vector Since it contains transduced T cells, the presence of replicable retroviruses (RCRs) in the blood of treated patients is also monitored.

[0247] If the subject is eligible for retreatment with KTE-C19, the final scan before retreatment is relevant to the retreatment. This is considered a baseline for evaluating responses.

[0248] If, at any point during the post-treatment evaluation period, the patient does not respond to the treatment (i.e., complete response or partial response) or progresses after responding, the patient will proceed directly to the 3-month follow-up appointment and long-term monitoring. The disease outcome will be monitored during the observation period.

[0249] Where appropriate, all subjects will be monitored for survival and disease status during the long-term follow-up period. Subjects will begin the long-term follow-up period after completing their 3-month post-treatment assessment (whether they responded to the treatment or proceeded directly to the 3-month examination due to disease progression). Starting from day 0 (KTE-C19 injection), subjects will return to the clinic every 3 months (±2 weeks) until month 18; every 6 months (±1 month) from month 24 to month 60; subjects will return to the clinic once a year starting from year 6, i.e., month 72 (±3 months), up to a maximum of 15 years. At this examination, the following will be observed: Complete the following steps: physical examination; PET-CT scan; disease assessment; clinical tests including CBC, anti-KTE-C19 antibody, lymphocyte subset, anti-CD19 CAR+ T cell, and RCR analysis with differential diagnosis; neurological and hematological tests. Reporting of target adverse events / serious adverse events up to disease progression (24 months or up to disease progression, whichever occurs first), including target, infection, autoimmune disorders, and secondary malignancies; description of target concomitant medications (up to 2 years after disease progression), including gamma globulin, immunosuppressants, anti-infectives, vaccines, and any treatments for the management of progressive disease.

[0250] The evaluation includes baseline PET-CT scans of the neck, chest, abdomen, and pelvis, along with appropriate imaging of all other sites of the disease. Subjects will undergo their first KTE-C19 post-infusion planned PET-CT tumor assessment 4 weeks after KTE-C19 injection, and will undergo the assessment at the regular intervals mentioned above.

[0251] Bone marrow aspirate and biopsy are performed on subjects who have been determined to have a complete response (CR). According to the revised IWG criteria for evaluating the effectiveness of treatment for malignant lymphoma, subjects who have myeloma spread of lymphoma prior to treatment Bone marrow aspirate and biopsy should only be performed if there is a clinical suspicion that lymphoma has spread to the bone marrow after the procedure, based on new abnormalities in peripheral blood count or blood smear. To assign a complete response (CR) to the procedure, the bone marrow aspirate and biopsy must not show morphological evidence of the disease, or if morphological uncertainty, immunohistochemical testing should be negative. It must be a sexual thing.

[0252] Test evaluation items major The primary endpoint for Phase 1 is the incidence of adverse events defined as dose-limiting toxicity (DLT). The primary endpoint for Phase 2 is the incidence of adverse events as determined by the principal investigator. According to the revised IWG criteria for evaluating the effectiveness of treatment for genital lymphoma, complete response or partial response This is the objective response rate (ORR), defined as the incidence rate of either of the two. By the analysis cutoff date, the customer All subjects who do not meet the criteria for observational response will be considered non-responders.

[0253] Secondary The objective response rate of the subjects in Phase 1 will be summarized. The objective response rate among the subjects in Phase 2 will be determined by the IRRC. The objective response rate will be the occurrence of either a complete response or a partial response according to the revised IWG treatment response criteria for malignant lymphoma, as determined by the IRRC. Defined as a rate. All subjects that do not meet the objective response criteria by the cutoff date of the analysis data are considered non-responders. Duration of response (DOR) for subjects who experienced an objective response. This refers to the revised IWG criteria for evaluating the effectiveness of treatment for malignant lymphoma, starting from the date of the first objective response. This is defined as disease progression due to the disease or death from an unrelated cause. Subjects who do not meet the criteria for progression or death by the cutoff date of the analysis data are censored on the final evaluable disease determination date, and their response is recorded as ongoing.

[0254] Dose-limiting toxicity (DLT) Dose-limiting toxicity is defined as the following KTE-C19-related events that occur within the first 30 days after KTE-C19 injection: a) Grade 4 neutropenia lasting longer than 21 days from the date of cell transplantation; b) Grade 4 thrombocytopenia lasting longer than 35 days from the date of cell transplantation; c) Any intubation requiring intubation, including Grade 4 confusion requiring intubation to secure the airway. KTE-C19-related adverse events are considered DLTs; d) All other Grade 3 toxic and DLT-not-considered conditions lasting longer than 3 days All Grade 4 toxicity excluding the following: (i) aphasia / language impairment or confusion / cognitive impairment that is Grade 1 or less within 2 weeks and recovers to baseline within 4 weeks; (ii) fever of Grade 3; (iii) lymphopenia, decreased hemoglobin, neutropenia, and thrombocytopenia that meet the above definition of DLT. Unless otherwise specified, myelosuppression as defined by neutropenia and thrombocytopenia (including bleeding in a platelet count of less than 50 × 10⁹ / L and bacterial infections recorded in a neutropenic state); (iv) immediate hypersensitivity reactions occurring within 2 hours of cell infusion (related to cell infusion) and reversible to grade 2 or less within 24 hours of cell administration with standard treatment; and (v) grade 3 or higher Or, it is hypogammaglobulinemia of type 4.

[0255] The CRS (Critical Risk Factor) is graded using the revised grading system (Lee 2014). Adverse events attributable to the CRS are mapped to the overall CRS grading determination for DLT (Disadvantageous Risk Factor) assessment.

[0256] During Phase 1, to evaluate the safety of the KTE-C19 scheme, patients with DLBCL, PMBCL, or TFL were tested. Approximately 6 to 24 subjects will be enrolled. Subjects in each cohort will be evaluated for DLT within the first 30 days after completion of each subject's KTE-C19 infusion. If the DLT incidence rate among the subjects is ≤1 out of 6, cohort B1 may be investigated, or the trial may proceed to phase 2 of a clinical trial. The decision will be based on overall benefit / risk and available biomarker data.

[0257] However, if two of the six registered subjects exhibit the protocol-defined DLT during Phase 1, the SRT will be administered to the additional two groups of three subjects at the same dose as the initial six subjects. It may be recommended to enroll a total of up to 12 participants. In this scenario, if ≤ 2 of the first 9 participants or ≤ 3 of the 12 participants show DLT, they can be moved to an additional cohort, or We will move on to Phase 2 of the trial.

[0258] If the DLT incidence rate for the target group is >2 / 6 people, >3 / 9 people, or >4 / 12 people, then additional treatment will be administered. Other KTE-C19 methods can be investigated in 6 to 12 subjects (Figure 3). The same DLT rules as above apply.

[0259] Example 4 The desired cell dose was achieved by transducing autologous lymphocytes with g-mouse retrovirus containing the anti-CD19 CAR construct gene, generating T cell products, and then proliferating them. After harvesting or co-culture with CD19+ cells, flow cytometry and multiplex sampling of the co-culture supernatant are performed. The characteristics of anti-CD19 CAR+ T cell products were evaluated by itokine analysis. CAR+ T cells were co-cultured with K562-CD19 cells or K562-NGFR control cells in an effector-to-target ratio of 1:1 for product characterization. The standard incubation time was 18 hours. Patients with relapsed / anti-treatment B-cell malignancies were conditioned with cyclophosphamide and fludarabine, and then given anti-CD19 CAR+ T cells.

[0260] Cytokine and chemokine levels were measured using the EMDmillipore Luminex® xMAP® multiplex assay. Data acquisition and analysis were performed using the Luminex 200® instrument and xPONENT® 3.1 data analysis software. Regarding IL-7, Using the Human IL-7 Quantikine HS ELISA Kit (HS750), follow the manufacturer's guidelines. The samples were used in a neat state. The number of circulating CAR T cells was measured by quantitative PCR analysis. 300 mg / m² on days -5 and -4. 2 Cyclophosphamide, as well as 30 mg / m² on days -5, -4, and -3. 2 A pre-conditioning regimen consisting of fludarabine was administered to the patient. Before administration of cyclophosphamide and fludarabine on days -12 to -5 ("pre"), and on day 0, CAR+ T Patient serum was collected immediately before ("after") cell administration and on selected days up to 18 days after CAR+ T cell administration. As shown in Figure 6, serum concentrations of GF-CSF, IL-2, MCP-1, IL-6, IL-10, MCP-4, CRP, IFN-gamma, granzyme A, IL-15, IL-5, granzyme B, IL-8, IP-10, MIP-1b, PLGF, IL-16, TARC, eotaxin-3, sICAM-1, sVCAM-1, and SAA were measured before and after conditioning and on selected days after CAR+ T cell administration. The concentration of certain cytokines was 300 mg / m². 2 Cyclophosphamide and 30 mg / m² 2 Fuldarabi It was found that the levels increased in patient serum after conditioning using (Figures 7A-7I). (Figures 7A-7D, 7G, 18A, and 18C-18E). In particular, the concentrations of IL-15, IL-7, PLGF, CRP, and MCP-1 significantly increased after conditioning with cyclophosphamide and fludarabine (Figures 7A-7D, 7G, 18A, and 18C-18E). Increases were also observed in the concentrations of IL-5, IL-10, IP-10, and s-ICAM1. (Figures 7E-7F, 7H-7I, and 18B). Conversely, perforin is cyclophosphamide and A decrease was found after conditioning with fludarabine (Figure 18F). As shown in Figure 18G, serum concentrations of various other analytes were observed to increase or decrease after preliminary conditioning. Additional patients were treated, and the results are shown in Figures 11–17. In addition, increased serum levels of IL-15 (Figure 19A) and IP-10 (Figure 19B), as well as decreased serum levels of perforin (Figure 19C), were found to be significantly correlated with positive objective responses in patients treated with CAR T cells after preliminary conditioning.

[0261] Peripheral blood lymphocytes (PBLs) and serum after CAR+ T cell infusion were subjected to flow cytometry. - and evaluated by multiple cytokine analysis. Site of anti-CD19 CAR+ T cells before injection Kine production was compared to K562-NGFR-negative controls (Figure 8). T1, T2, and immunoconstitutive cytokines GM-CSF, IL-2, IFN-gamma, IL-5, IL-4, and IL-13, as well as pro-inflammatory cytokines and chemokines TNF-alpha, IL-6, granzyme B, MIP-1b (beta), MIP-1a (a) The concentrations of rufa and sCD137 were higher in the anti-CD19 CAR+ T cell sample than in the negative control. (Figures 8A-8L). In addition, the association of target antigens by product T cells before injection regulates their activity. Upregulators for receptors such as CD107a (Alpha), 401BB, and PD-1 that can be used. It provides rations (Figures 9A-9C).

[0262] Using BD FACSCanto II with FlowJo software for data acquisition and analysis. Multicolor flow cytometry was performed. A shorter manufacturing process yielded CAR+ T cell products that more highly expressed CD4+, naive, and central memory T cells (Figure 10). After injection, CAR+ T cells are mainly differentiated T cells and some central memory or nitric acid. It shows a diversified subset composition including β T cells (Figure 10).

[0263] Anti-CD19 CD28 zetaCAR+ T cells are clinically effective in both lymphoma and leukemia. It induces a long-lasting response. Sustained clinical responses can occur in circulation without long-lasting CAR+ T cells and restore normal B cells. Cyclophosphamide and fludara Conditioning using bottles is advantageous for the homeostatic proliferation, activation, and transport of T cells. The immune environment is modified by inducing molecules that may be involved. CAR+ T cell treatment should be performed within 3 weeks after treatment. This leads to a rapid increase in circulating cytokines and chemokines, followed by their recovery.

[0264] Example 5 300 mg / m² 2 The above cyclophosphamide and 30 mg / m² 2 The above consists of fludarabine doses. A study will be conducted to test the safety and efficacy of treating subjects using a non-myeloablative conditioning method. Further induction of lymphocyte depletion will be achieved through in vivo proliferation of KTE-C19. These conditioning chemotherapy agents are used in dosages to create a suitable environment.

[0265] The registered subjects were leukocyte ferrets used to obtain PBMCs for the production of anti-CD19 CAR+ T cells. The patient will receive the next dose of 500 mg / m³ administered on days -5 to -3. 2 Cyclophosphamide / day and 60 mg / m² 2 The patient receives conditioning chemotherapy containing fludarabine daily. Next, on day 0, the patient receives IV administration of anti-CD19 CAR+ T cells / kg. The starting dose is as follows: This is anti-CD19 CAR+ T cell 2×10 6 The patient may receive a dose of 1 / kg (±20%), after which the starting dose may be increased or decreased depending on the patient's response.

[0266] Conditioning chemotherapy for anti-CD19 CAR+ T cells and adverse effects, serum saturation, etc. after administration. Monitor subjects for itokine levels, T cell count, and disease response. (No limitations) IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-15, IL-16, IL-21, MCP-1, IP-10, PLGF, sICAM-1, CRP, VEGF, VEGF-C, VEGF-D, sVCAM-1, MIP-1β, FGF2, IL-1b, Otaxin, GM-CSF, IFN-gamma, IL-12p40, MDC, IL-12p70, IL-13, IL-17A, MIP-1a, TNFa, TNFb, Granzyme A, Granzyme B, Perforin, SAA, MCP-4, and TARC Serum levels of various cytokines, chemokines, effectors, inflammatory markers, and adhesion molecules, including cyclophosphamide, fludarabine, and anti-CD19 CAR+ T, are measured before and after conditioning to determine the effectiveness of conditioning chemotherapy. Cells are collected before or after each administration and all levels are compared to levels before conditioning chemotherapy. Disease response is compared to the cytokine profile of each patient after conditioning to assess disease response and one or more sites after conditioning. Check for any correlation with Cain's level.

[0267] The occurrence of adverse effects will be closely monitored to determine the maximum tolerable doses of cyclophosphamide and fludarabine. Adverse effects may be medically controlled as needed. The doses of one or both cyclophosphamide and fludarabine may be increased or decreased to improve clinical efficacy and limit adverse effects. Any subject showing disease progression following an initial partial response may receive a second treatment with the same or different levels of cyclophosphamide and / or fludarabine. Throughout this application, various publications have been made, with the author's name and date in parentheses. Alternatively, they may be referenced by patent number or patent publication number. Complete citations of these publications may be found at the end of the specification immediately preceding the claims. The disclosures of these publications are incorporated into this application by reference in their entirety to better describe the current state of the art as known to those skilled in the art as of the date of invention described and claimed herein. However, the citation of references herein should not be construed as an acknowledgment that such references are prior art of the present invention. All of the various aspects, embodiments, and options described herein can be combined in any and all variations.

[0268] Example 6 For preconditioning using one or more preconditioning agents To predict the potential patient response to T-cell therapy based on cytokine responsiveness A trial will be conducted. For this trial, cancer patients who are suitable for T-cell therapy will be selected.

[0269] Collect the patient's blood before any intervention. Then, one or more preliminary cells according to the present invention Pre-conditioning is performed by administering conditioning agents to the patient. For example, the patient receives 300 or 500 mg / m² prior to T-cell therapy. 2 2 or 3 doses of cyclophosphamide per day Daily doses of 30 or 60 mg / m² 2 Treatment may involve 3, 4, or 5 days of fludarabine per day. After preliminary conditioning, immediately before administering T-cell therapy, for example on the same day as administration, the patient Blood is collected again. Next, the patient is given T-cell therapy to improve disease response, for example, in progressive diseases. Monitor for partial or complete response.

[0270] Non-limitingly, a variety of cytokines including IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. Serum levels of itokines are analyzed by analyzing blood collected before and after preliminary conditioning. The multiplier of change in each cytokine is recorded for each patient and compared to the overall disease response. An increase or decrease in any one or more cytokines indicates an overall disease response. A correlation study will be conducted to determine whether it predicts sex.

[0271] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being incorporated by reference. However, the reference of any reference herein should not be construed as an acknowledgment that such reference is prior art of the present invention.

[0272] While the present invention has been described in general terms, further understanding can be gained by referring to the examples provided herein. These examples are for illustrative purposes only and are not intended to limit the invention. Item 1 (i) The step of administering to the patient one or more preconditioning agents capable of increasing serum levels of at least one additional cytokine selected from the group consisting of interleukin-15 ("IL-15"), interleukin-7 ("IL-7"), and monocyte chemotactic protein 1 ("MCP-1"), C-reactive protein ("CRP"), placental growth factor ("PLGF"), interferon-gamma-inducible protein 10 ("IP-10"), and any combination thereof, and (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. A method for treating cancer in a patient suitable for the T-cell therapy, including the following: Section 2 A method for treating cancer in a patient suitable for T-cell therapy, comprising the step of administering to the patient one or more preconditioning agents capable of increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, wherein the patient is treated with T-cell therapy if he or she exhibits increased serum levels of IL-15, IL-7, and at least one additional cytokine. Section 3 (i) The step of administering to the patient one or more preconditioning agents capable of increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (ii) a step of administering an additional amount of one or more preconditioning agents, or a step of administering an effective amount of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (iii) If the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine, the patient is given T-cell therapy. A method for treating cancer in a patient suitable for the T-cell therapy, including the following: Section 4 A method for identifying a patient suitable for T-cell therapy, comprising the step of administering to the patient one or more preconditioning agents capable of increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof. Section 5 (i) The step of administering to the patient one or more preconditioning agents capable of increasing serum levels of at least one additional cytokine selected from the group consisting of IL-15, IL-7, and MCP-1, CRP, PLGF, IP-10, and any combination thereof, and (ii) The stage in which T-cell therapy is administered when the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. A method for identifying a patient suitable for the T-cell therapy, including the following: Section 6 (i) The step of administering to the patient one or more preconditioning agents capable of increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (ii) a step of administering an additional amount of one or more preconditioning agents, or a step of administering an effective amount of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (iii) If the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine, the patient is given T-cell therapy. A method for identifying patients suitable for the T-cell therapy, including the following. Section 7 The stage of administering one or more preliminary conditioning agents to the patient. A method for preconditioning a patient requiring T-cell therapy, comprising increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, wherein the patient is treated with the T-cell therapy when he or she shows increased serum levels of IL-15, IL-7, and at least one additional cytokine. Section 8 A method for preconditioning patients requiring T-cell therapy by increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (i) the step of administering one or more preconditioning agents to the patient, (ii) The step of administering the T-cell therapy if the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. Methods that include... Section 9 A method for preconditioning patients requiring T-cell therapy by increasing serum levels of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (i) The step of administering one or more preconditioning agents to the patient, (ii) a step of administering an additional amount of one or more preconditioning agents, or a step of administering an effective amount of IL-15, IL-7, and at least one additional cytokine selected from the group consisting of MCP-1, CRP, PLGF, IP-10, and any combination thereof, (iii) The step of administering the T-cell therapy if the patient shows elevated serum levels of IL-15, IL-7, and at least one additional cytokine. Methods that include... Item 10 The method according to any one of claims 1 to 9, further comprising the step of measuring serum levels of IL-15, IL-7, and at least one cytokine after administration of one or more preconditioning agents. Section 11 The method according to any one of claims 1 to 10, wherein one or more preconditioning agents reduce the number of endogenous lymphocytes in a patient. Section 12 The method according to item 11, wherein the endogenous lymphocytes include regulatory T cells, B cells, natural killer cells, CD4+ T cells, CD8+ T cells, or any combination thereof. Section 13 The method according to any one of claims 1 to 12, wherein the serum level of IL-7 in the patient increases by at least 2, at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 60, at least 70, at least 80, or at least 90 times after administration of one or more preconditioning agents compared to the serum level of IL-7 before administration of one or more preconditioning agents. Section 14 The method according to any one of claims 1 to 13, wherein the serum level of IL-15 in the patient increases by at least 5 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, or at least 90 times after administration of one or more preconditioning agents compared to the serum level of IL-15 before administration of one or more preconditioning agents. Item 15 The method according to any one of claims 1 to 14, wherein the serum level of MCP-1 in the patient increases by at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 20 times after administration of one or more preconditioning agents compared to the serum level of MCP-1 before administration. Item 16 The method according to any one of claims 1 to 15, wherein the serum level of PLGF in the patient increases by at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, or at least 100 times after administration of one or more preconditioning agents compared to the serum level of PLGF before administration of one or more preconditioning agents. Item 17 The method according to any one of claims 1 to 16, wherein the serum level of CRP in a patient increases by at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least about 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, or at least 100 times after administration of one or more preconditioning agents compared to the serum level of CRP before administration of one or more preconditioning agents. Section 18 The method according to any one of claims 1 to 17, wherein the serum level of IP-10 in the patient increases by at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, or at least 30 times after administration of one or more preconditioning agents compared to the serum level of IP-10 before administration of one or more preconditioning agents. Section 19 The method according to any one of paragraphs 1 to 18, wherein the preconditioning agent further increases serum levels of interleukin-10 ("IL-10"), interleukin-5 ("IL-5"), interleukin-8 ("IL-8"), soluble intercellular adhesion molecule 1 ("sICAM-1"), soluble vascular adhesion molecule 1 ("sVCAM-1"), or any combination thereof in the patient. Section 20 The method according to paragraph 19, wherein the serum level of IL-10 in the patient increases by at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, or at least 20 times after administration of one or more preconditioning agents compared to the serum level of IL-10 before administration. Section 21 The method according to claim 19 or 20, wherein the serum level of IL-5 in the patient increases by at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 times after administration of one or more preconditioning agents compared to the serum level of IL-5 before administration of the preconditioning agent. Section 22 The method according to any one of claims 19 to 21, wherein the serum level of IL-8 in the patient increases by at least 2 times, at least 5 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, or at least 100 times after administration of one or more preconditioning agents compared to the serum level of IL-8 before administration of one or more preconditioning agents. Section 23 The method according to any one of claims 19 to 22, wherein the serum level of sICAM-1 in the patient increases by at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, or at least 30 times after administration of one or more preconditioning agents compared to the serum level of sICAM-1 before administration of one or more preconditioning agents. Section 24 The method according to any one of claims 19 to 23, wherein the serum level of sVCAM-1 in the patient increases by at least 1.5 times, at least 2 times, at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, at least 4.5 times, or at least 5 times after administration of one or more preconditioning agents compared to the serum level of sVCAM-1 before administration of one or more preconditioning agents. Section 25 The method according to any one of claims 1 to 24, wherein one or more preconditioning agents comprises an alkylating agent. Section 26 The method according to claim 24, wherein the alkylating agent is selected from the group consisting of melphalan, chlorambucil, cyclophosphamide, mechloretamine, mustine (HN2), uramustine, uracil mustard, melphalan, chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, streptozosin, alkyl sulfonates, busulfan, thiotepa or their analogues, and any combination thereof. Section 27 The method according to item 26, wherein one or more preconditioning agents are platinum-based preconditioning agents. Section 28 The method according to item 27, wherein the platinum-based preconditioning agent is selected from the group consisting of platinum, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, procarbazine, altoretamine, triazene, dacarbazine, mitozolomid, temozolomid, dacarbazine, temozolomid, and any combination thereof. Section 29 The method according to any one of claims 1 to 26, wherein one or more preconditioning agents are cyclophosphamide. Item 30 The method according to any one of claims 1 to 29, wherein one or more preconditioning agents are purine analogs. Section 31 The method according to claim 30, wherein the purine analog is selected from the group consisting of azathioprine, 6-mercaptopurine, mercaptopurine, thiopurine, thioguanine, fludarabine, pentostatin, cladribine, and any combination thereof. Section 32 The method according to any one of claims 1 to 24, wherein one or more preconditioning agents are cyclophosphamide and purine analogs. Item 33 The method according to claim 32, wherein the purine analog is selected from the group consisting of azathioprine, 6-mercaptopurine, mercaptopurine, thiopurine, thioguanine, fludarabine, pentostatin, cladribine, and any combination thereof. Section 34 The method according to claim 32, wherein one or more preconditioning agents are cyclophosphamide and pentostatin. Section 35 The method according to item 32, wherein one or more preconditioning agents are cyclophosphamide and fludarabine. Section 36 The effective dose of cyclophosphamide is approximately 300 mg / m². 2 / day~about 2000mg / m 2 The method described in paragraphs 32-35 is / day. Section 37 The effective dose of cyclophosphamide is 300 mg / m². 2 Higher than / day, 2000 mg / m² 2 A method lower than / day, as described in items 32-35. Section 38 The effective dose of fludarabine is approximately 20 mg / m². 2 / day~about 900mg / m 2 The method described in any one of paragraphs 35 to 37, which is / day. Item 39 The effective dose of fludarabine is 30 mg / m². 2 Higher than / day, 900mg / m² 2 A method lower than / day, as described in any one of items 35-37. Section 40 The effective dose of cyclophosphamide is approximately 350 mg / m². 2 / day~about 2000mg / m 2 / day, at least approximately 400 mg / m² 2 / day~about 2000mg / m 2 / day, approximately 450mg / m 2 / day~about 2000mg / m 2 / day, about 500mg / m 2 / day~about 2000mg / m 2 / day, about 550mg / m 2 / day~about 2000mg / m 2 / day, or approximately 600 mg / m² 2 / day~about 2000mg / m 2 The method described in any one of paragraphs 32 to 39, which is / day. Section 41 The effective dose of cyclophosphamide is approximately 350 mg / m². 2 / day~about 1500mg / m 2 / day, about 350mg / m 2 / day~about 1000mg / m 2 / day, about 400mg / m 2 / day~about 900mg / m 2 / day, approximately 450mg / m 2 / day~about 800mg / m 2 / day, approximately 450mg / m 2 / day~about 700mg / m 2 / day, about 500mg / m 2 / day~about 600mg / m 2 / day, or approximately 300 mg / m² 2 / day~about 500mg / m 2 The method described in any one of paragraphs 32 to 39, which is / day. Section 42 The effective dose of cyclophosphamide is approximately 350 mg / m². 2 / day, about 400mg / m 2 / day, about 450mg / m 2 / day, about 500mg / m 2 / day, about 550mg / m 2 / day, about 600mg / m 2 / day, approximately 650mg / m 2 / day, about 700mg / m 2 / day, about 800mg / m 2 / day, approximately 900mg / m 2 / day, or approximately 1000 mg / m² 2 The method described in paragraph 41, which is / day. Section 43 The effective dose of fludarabine is approximately 35 mg / m². 2 / day~about 900mg / m 2 / day, about 40mg / m 2 / day~about 900mg / m 2 / day, about 45mg / m 2 / day~about 900mg / m 2 / day, about 50mg / m 2 / day~about 900mg / m 2 / day, about 55mg / m 2 / day~about 900mg / m 2 / day, or approximately 60mg / m² 2 / day~about 900mg / m 2 The method described in any one of paragraphs 35 to 42, which is / day. Section 44 The effective dose of fludarabine is approximately 35 mg / m².2 / day~about 900mg / m 2 / day, about 35mg / m 2 / day~about 800mg / m 2 / day, about 35mg / m 2 / day~about 700mg / m 2 / day, about 35mg / m 2 / day~about 600mg / m 2 / day, about 35mg / m 2 / day~about 500mg / m 2 / day, about 35mg / m 2 / day ~ approx. 400mg / m 2 / day, about 35mg / m 2 / day ~ approx. 300mg / m 2 / day, about 35mg / m 2 / day~about 200mg / m 2 / day, about 35mg / m 2 / day~about 100mg / m 2 / day, about 40mg / m 2 / day~about 90mg / m 2 / day, about 45mg / m 2 / day~about 80mg / m 2 / day, about 45mg / m 2 / day~about 70mg / m 2 / day, or approximately 50 mg / m² 2 / day~about 60mg / m 2 The method described in any one of paragraphs 35 to 42, which is / day. Section 45 The effective dose of fludarabine is approximately 35 mg / m². 2 / day, about 40mg / m 2 / day, about 45mg / m 2 / day, about 50mg / m 2 / day, about 55mg / m 2 / day, about 60mg / m 2 / day, about 65mg / m 2 / day, about 70mg / m 2 / day, about 75mg / m 2 / day, about 80mg / m 2 / day, about 85mg / m 2 / day, approximately 90mg / m 2 / day, approximately 95mg / m 2 / day, about 100mg / m 2 / day, about 200mg / m 2 / day, or approximately 300 mg / m² 2 The method described in paragraph 44, which is / day. Section 46 The effective dose of cyclophosphamide is approximately 500 mg / m². 2 The effective dose of fludarabine is 60 mg / m² / day. 2 The method described in paragraph 35, which is / day. Section 47 The method according to any one of claims 1 to 46, wherein one or more preconditioning agents are administered daily for at least one day, at least two days, at least three days, at least four days, at least five days, at least six days, or at least seven days. Section 48 The method according to any one of items 1 to 46, wherein one or more pre-conditioning agents are administered daily for about three days. Section 49 The method described in any one of paragraphs 35 to 48, wherein cyclophosphamide is administered before, after, or concurrently with fludarabine. Item 50 The method described in paragraph 49, wherein cyclophosphamide is administered before fludarabine. Section 51 The method according to any one of claims 1 to 50, further comprising the step of administering one or more doses of IL-2. Section 52 The method according to paragraph 51, wherein each dose of IL-2 is at least about 10,000 IU / kg, at least about 50,000 IU / kg, at least about 100,000 IU / kg, at least about 200,000 IU / kg, at least about 400,000 IU / kg, at least about 600,000 IU / kg, at least about 700,000 IU / kg, at least about 800,000 IU / kg, or at least about 1,000,000 IU / kg. Section 53 The method according to any one of items 1 to 52, wherein the T cell therapy is selected from the group consisting of tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, modified autologous cell therapy (eACT), allogeneic T cell transplantation, and any combination thereof. Section 54 The method according to any one of claims 1 to 53, further comprising the step of collecting blood cells from a patient before administering one or more preconditioning agents. Section 55 The method according to paragraph 54, further comprising the step of manipulating blood cells to express a chimeric antigen receptor ("manipulated CAR cells") or a T cell receptor ("manipulated TCR cells"). Section 56 A method according to any one of items 1 to 55, wherein the T cell therapy includes engineered CAR cell therapy or engineered TCR cell therapy. Section 57 The method described in paragraph 56, wherein manipulated CAR cells or manipulated TCR cells are used to treat a tumor in a patient. Section 58 The method according to any one of claims 1 to 57, wherein the administration of one or more preconditioning agents begins at least 7 days, at least 6 days, at least 5 days, at least 4 days, at least 3 days, at least 2 days, or at least 1 day before the implementation of T-cell therapy. Section 59 The method according to paragraph 58, wherein the administration of a first pre-conditioning agent begins approximately 7 days before the administration of T-cell therapy, and the administration of a second pre-conditioning agent begins approximately 5 days before the administration of T-cell therapy. Section 60 The method according to paragraph 58, wherein a first pre-conditioning agent is administered to the patient for two days, approximately seven days and approximately six days before the administration of T-cell therapy. Section 61 The method according to paragraph 60, wherein a second pre-conditioning agent is administered to the patient for 5 days about 5, 4, 3, 2, and 1 day before the administration of T-cell therapy. Section 62 The method described in any one of paragraphs 1 to 57, wherein the administration of one or more preconditioning agents begins approximately five days before the administration of T-cell therapy. Section 63 The method according to paragraph 62, wherein a first pre-conditioning agent is administered to the patient for three days about five, four, and three days before the administration of T-cell therapy. Section 64 The method according to any one of paragraphs 1 to 63, wherein the administration of one or more preconditioning agents induces improved antitumor efficacy of T-cell therapy compared to the antitumor efficacy of T-cell therapy without the administration of one or more preconditioning agents. Section 65 Patients who have received one or more preconditioning agents and T-cell therapy have received treatment for IL-15, IL-7, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, IL-1, IL-2, IL-3, IL-4, IL-6, IL-9, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-20, and granulocyte-macrophage cells. Knee-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor D (VEGF-D), macrophage inflammatory protein 1β (MIP-1β), leukemia suppressor factor (LIF), oncostatin M (OSM), interferon (IFN) alpha, IFN-beta, IFN-gamma, tumor necrosis factor (TNF) alpha, TNF-beta, CD154, lymphotoxin (LT) beta, 4-1BB ligand (4-1 A method according to any one of items 1 to 64, which shows increased serum levels of cytokines or pro-inflammatory factors selected from the group consisting of BBL), proliferation-inducing ligand (APRIL), CD70, CD153, CD178, glucocorticoid-inducing TNFR-related ligand (GITRL), tumor necrosis factor superfamily member 14 (TNFSF14), OX40L, TNF and ApoL-related leukocyte expression ligand 1 (TALL-1), TNF-related apoptosis-inducing ligand (TRAIL), chemokine (CC motif) ligand (CCL) 1, macrophage inflammatory protein 1 alpha (MIP-1a or CCL3), CCL5, monocyte-specific chemokine 3 (MCP3 or CCL7), monocyte chemotactic protein 2 (MCP-2 or CCL8), CCL13, thymic and activation-regulating chemokines (TARC or CCL17), CCL22, and any combination thereof. Section 66 The method according to any one of claims 1 to 65, wherein two or more preconditioning agents are administered simultaneously or sequentially. Section 67 The method according to paragraph 66, wherein a first pre-conditioning agent is administered to the patient before or after a second pre-conditioning agent. Section 68 The method described in any one of sections 55 to 67, wherein the manipulated CAR T cells express a chimeric antigen receptor. Section 69 The method according to item 68, wherein the chimeric antigen receptor comprises a binding molecule to a tumor antigen. Section 70 The method according to item 69, wherein the binding molecule is an antibody or its antigen-binding molecule. Section 71 The method according to item 70, wherein the binding molecule is an antigen-binding molecule selected from the group consisting of scFv, Fab, Fab', Fv, F(ab')2, dAb, and any combination thereof. Section 72 The method according to any one of items 68 to 71, wherein the chimeric antigen receptor includes a hinge region. Section 73 The method according to item 72, wherein the hinge region is IgG1, IgG2, IgG3, IgG4, IgA, IgD, IgE, IgM, CD28, or CD8 alpha. Section 74 The method according to item 73, wherein the hinge region is of IgG4. Section 75 The method according to any one of items 68 to 74, wherein the chimeric antigen receptor includes a transmembrane domain. Section 76 The method according to item 75, wherein the transmembrane domain is a transmembrane domain derived from CD28, CD8 alpha, CD4, or CD19. Section 77 The method according to item 76, wherein the transmembrane domain is the transmembrane domain of CD28. Section 78 The method according to any one of claims 68 to 77, wherein the chimeric antigen receptor further comprises a co-stimulatory signaling region. Section 79 The method according to item 78, wherein the co-stimulatory signaling region is a signaling region derived from CD28, OX-40, 4-1BB, CD27, inducible T cell costimulatory molecule (ICOS), CD3 gamma, CD3 delta, CD3 epsilon, CD247, Ig alpha (CD79a), or an Fc gamma receptor. Section 80 The method according to item 79, wherein the co-stimulatory signaling region is the signaling region of CD28. Section 81 The method according to any one of items 68 to 80, wherein the chimeric antigen receptor further comprises a CD3 zeta signaling domain. Section 82 Tumor antigens include CD19, CD20, type 1 receptor tyrosine kinase-like orphan receptor ("ROR1"), CD22, carcinoembryonic antigen, alpha-fetoprotein, CA-125, 5T4, mucin 1 ("MUC-1"), epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, variant p53, variant ras, HER2 / Neu, folate-binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, C A method according to any one of claims 69 to 81, selected from the group consisting of D123, CD33, CD138, CD23, CD30, CD56, c-Met, mesotheline, GD3, HERV-K, IL-IIR alpha, kappa chain, lambda chain, chondroitin sulfate proteoglycan ("CSPG4"), ERBB2, EGFRvIII, VEGFR2, a combination of HER2-HER3, a combination of HER1-HER2, and any combination thereof. Section 83 The method described in any one of paragraphs 68 to 82, wherein manipulated CAR cells reduce the size of the tumor. Section 84 The method according to any one of items 55 to 67, wherein the manipulated TCR cells express a T cell receptor. Section 85 The method according to item 84, wherein the T cell receptor contains a binding molecule for a tumor antigen. Section 86 The method according to item 85, wherein the tumor antigen is selected from the group consisting of CD19, CD20, ROR1, CD22, carcinoembryonic antigen, alpha-fetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, variant p53, variant ras, HER2 / Neu, folate-binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c-Met, mesothelin, GD3, HERV-K, IL-IIR alpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, combinations of HER2-HER3, combinations of HER1-HER2, and any combination thereof. Section 87 The method according to item 84, wherein the T cell receptor contains a molecule that binds to a viral oncogene. Section 88 The method according to item 87, wherein the viral oncogene is selected from human papillomavirus (HPV), Epstein-Barr virus (EBV), and human T-lymphotropic virus (HTLV). Section 89 The method according to item 84, wherein the T cell receptor comprises a binding molecule to a testicular, placental, or fetal tumor antigen. Section 90 The method described in paragraph 89, wherein the testicular, placental, or fetal cancer antigen is selected from NY-ESO-1, synovial sarcoma X breakpoint 2 (SSX2), and melanoma antigen (MAGE). Section 91 The method according to item 84, wherein the T cell receptor contains a binding molecule for a lineage-specific antigen. Section 92 The method according to item 91, wherein the lineage-specific antigen is selected from melanoma antigen 1 (MART-1), gp100, prostate-specific antigen (PSA), prostate-specific membrane antigen (PSMA), and prostate stem cell antigen (PSCA), which are recognized by T cells. Section 93 The method according to any one of paragraphs 55-67 and 84-92, wherein manipulated TCR cells reduce the size of tumors in a patient. Section 94 The therapeutically effective dose of manipulated CAR cells or manipulated TCR cells is at least about 10 4 pieces, at least about 10 5 pieces, at least about 10 6 pieces, at least about 10 7 pieces, at least about 10 8 pieces, at least about 10 9 one, or at least about 10 10 The method described in any one of items 55 to 93, which is one item. Section 95 The therapeutically effective dose of manipulated CAR cells is approximately 10 4 pieces, about 105 pieces, about 10 6 pieces, about 10 7 pieces, about 10 8 pieces, about 10 9 10, or about 10 10 The method described in any one of items 55 to 93, which is one item. Section 96 The therapeutically effective dose of manipulated CAR cells is approximately 2 × 10⁻⁶ 6 pieces / kg, approximately 3×10 6 pieces / kg, approximately 4×10 6 pieces / kg, approximately 5×10 6 pieces / kg, approximately 6×10 6 pieces / kg, approximately 7×10 6 pieces / kg, approximately 8×10 6 pieces / kg, approximately 9×10 6 pieces / kg, approximately 1×10 7 pieces / kg, approximately 2×10 7 pieces / kg, approximately 3×10 7 pieces / kg, approximately 4×10 7 pieces / kg, approximately 5×10 7 pieces / kg, approximately 6×10 7 pieces / kg, approximately 7×10 7 pieces / kg, approximately 8×10 7 pieces / kg, or approximately 9 x 10 7 The method described in any one of items 55 to 93, wherein the amount is pieces / kg. Section 97 Tumors include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute The method according to item 83 or 93, selected from tumors resulting from lymphoblastic leukemia, chronic lymphocytic leukemia, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environment-induced cancers including asbestos-induced cancers, and combinations of the said cancers. Section 98 The method described in any one of paragraphs 69 to 97, wherein the tumor antigen is CD19. Section 99 The method described in paragraph 98, wherein the tumor is lymphoma or leukemia. Item 100 Lymphoma or leukemia includes B-cell chronic lymphocytic leukemia / small cell lymphoma, B-cell prelymphocytic leukemia, lymphoplasmacytic lymphoma (e.g., Waldenström macroglobulinemia), splenic marginal lamina lymphoma, hairy cell leukemia, plasmacytic neoplasms (e.g., plasmacytic myeloma (i.e., multiple myeloma), or plasmacytoma), extranodal marginal zone B-cell lymphoma (e.g., MALT). Lymphoma, Nodal marginal zone B-cell lymphoma, Follicular lymphoma, Transformed follicular lymphoma, Primary cutaneous follicular central lymphoma, Mantle cell lymphoma, Diffuse large B-cell lymphoma (DLBCL), Epstein-Barr virus positive DLBCL, Lymphomatoid granulomatosis, Mediastinal (thymic) large B-cell lymphoma, Intravascular large B-cell lymphoma, ALK+ large B-cell lymphoma Lymphoma, plasmablastic lymphoma, primary exudative lymphoma, large B-cell lymphoma occurring in HHV8-associated multicentric Castleman disease, Burkitt lymphoma / leukemia, T-cell pre-lymphocytic leukemia, T-cell macrogranular lymphocytic leukemia, aggressive NK-cell leukemia, adult T-cell leukemia / lymphoma, extranodal NK / T-cell lymphoma, enteropathy-associated T-cell lymphoma, hepatosplenic T-cell lymphoma, blastic NK-cell lymphoma The method described in paragraph 99, selected from the group consisting of lymphoma, mycosis fungoides / Sézary syndrome, primary cutaneous anaplastic large cell lymphoma, lymphomatoid papular dysplasia, peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, B-lymphoblastic leukemia / lymphoma, B-lymphoblastic leukemia / lymphoma with recurrent genetic abnormalities, T-cell lymphoblastic leukemia / lymphoma, and Hodgkin lymphoma. Section 101 The method according to any one of items 1 to 100, further comprising the step of administering physiological saline to the patient. Section 102 The method according to paragraph 101, wherein physiological saline is administered before the administration of one or more preconditioning agents. Section 103 The method according to any one of items 1 to 102, further comprising the step of administering mesna (sodium 2-mercaptoethanesulfonate) to the patient. Section 104 The method according to paragraph 103, wherein mesna is administered before the administration of one or more preconditioning agents. Section 105 The effective dose of cyclophosphamide is approximately 200 mg / m². 2 The method described in paragraph 35, which is / day. Section 106 The effective dose of cyclophosphamide is 200 mg / m². 2 / day~3000mg / m 2 The method according to paragraph 35, wherein the patient exhibits increased serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, or decreased serum levels of perforin and / or MIP-1b after administration of cyclophosphamide and fludarabine. Section 107 The effective dose of cyclophosphamide is 1000 mg / m². 2 / day~2000mg / m 2 The method described in any one of paragraphs 35-39, 43-45, 47-104, and 106, which is / day. Section 108 The effective dose of cyclophosphamide is approximately 1110 mg / m². 2 The method described in paragraphs 35-41, 43-45, 47-104, 106, and 107, which is / day. Section 109 The effective dose of cyclophosphamide is approximately 300 mg / m². 2 The method described in any one of paragraphs 35-39, 47-104, 106, and 107, which is / day. Section 110 The method described in any one of items 35-41, 43-45, 47-104, 106, and 107, wherein the effective dose of cyclophosphamide is approximately 30 mg / kg / day. Section 111 The effective dose of fludarabine is approximately 20 mg / m². 2 The method described in any one of paragraphs 35-42 and 47-110, which is / day. Section 112 The effective dose of fludarabine is approximately 25 mg / m². 2 The method described in any one of paragraphs 35-42 and 47-110, which is / day. Section 113 The effective dose of fludarabine is approximately 30 mg / m². 2 The method described in any one of paragraphs 35-42 and 47-110, which is / day. Section 114 The effective dose of cyclophosphamide is approximately 200 mg / m². 2 The effective dose of fludarabine is approximately 20 mg / m² / day. 2 The method described in any one of paragraphs 35-39 and 47-104, which is / day. Section 115 The effective dose of cyclophosphamide is approximately 300 mg / m². 2 The effective dose of fludarabine is approximately 30 mg / m² / day. 2 The method described in any one of paragraphs 35-39 and 47-104, which is / day. Section 116 The effective dose of cyclophosphamide is approximately 300 mg / m². 2 The effective dose of fludarabine is approximately 60 mg / m² / day. 2 The method described in any one of paragraphs 35-39 and 47-104, which is / day. Section 117 The effective dose of cyclophosphamide is approximately 500 mg / m². 2 The effective dose of fludarabine is approximately 30 mg / m² / day. 2 The method described in any one of paragraphs 35-41 and 47-104, which is / day. Section 118 The effective dose of cyclophosphamide is approximately 1110 mg / m². 2 The effective dose of fludarabine is approximately 25 mg / m² / day. 2 The method described in any one of paragraphs 35-41, 47-104, and 106, which is / day. Section 119 The effective dose of cyclophosphamide is approximately 2220 mg / m². 2 The effective dose of fludarabine is approximately 25 mg / m² / day. 2 The method according to paragraph 106, wherein the patient exhibits increased serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, or decreased serum levels of perforin and / or MIP-1b after administration of cyclophosphamide and fludarabine. Section 120 The therapeutically effective dose of manipulated CAR T cells is approximately 1.0 × 10⁶ cells. 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 2.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 3.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 4.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 5.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 6.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 7.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 8.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 9.0×10 5 pieces / kg ~ approx. 2×10 8 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 3×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 4×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 5×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 6×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 7×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 8×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 9×10 6 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 1×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 2×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 3×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 4×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 5×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 6×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 7×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 8×10 7 pieces / kg ~ approx. 9×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 8×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 7×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 6×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 5×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 4×107 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 3×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 2×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 1×10 7 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 9×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 8×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 7×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 6×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 5×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 4×10 6 pieces / kg, approximately 2×10 6 pieces / kg ~ approx. 3×10 6 pieces / kg, approximately 3×10 6 pieces / kg ~ approx. 8×10 7 pieces / kg, approximately 4×10 6 pieces / kg ~ approx. 7×10 7 pieces / kg, approximately 5×10 6 pieces / kg ~ approx. 6×10 7 pieces / kg, approximately 6×10 6 pieces / kg ~ approx. 5×10 7 pieces / kg, approximately 7×10 6 pieces / kg ~ approx. 4×10 7 pieces / kg, approximately 8×10 6 pieces / kg ~ approx. 3×10 7 pieces / kg, or approximately 9 x 10 6 pieces / kg ~ approx. 2×10 7 The method according to any one of items 55-93 and 97-116, wherein the amount is pieces / kg. Section 121 (i) The step of administering one or more preliminary conditioning agents to the patient, (ii) A step of measuring serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, perforin, MIP-1b, or any combination thereof, and (iii) Characterizing the one or more preconditioning agents as effective for preparing a subject for T-cell therapy if, after administration of the one or more preconditioning agents, the subject exhibits elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, e.g., elevated serum levels of IL-15, IP-10, and / or IL-7, and / or decreased serum levels of perforin and / or MIP-1b. A method for identifying the dose of one or more pre-conditioning agents effective for preparing a subject for the T-cell therapy, including the following: Section 122 (i) The step of administering one or more preliminary conditioning agents to the patient, (ii) A step of measuring serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, perforin, MIP-1b, or any combination thereof, and (iii) Characterizing the one or more preconditioning agents as effective for preparing a subject for T-cell therapy if, after administration of the one or more preconditioning agents, the subject exhibits elevated serum levels of IL-7, IL-15, IL-10, IL-5, IP-10, IL-8, MCP-1, PLGF, CRP, sICAM-1, sVCAM-1, or any combination thereof, e.g., elevated serum levels of IL-15, IP-10, and / or IL-7, and / or decreased serum levels of perforin and / or MIP-1b. A method for verifying the efficacy of one or more preliminary conditioning agents for preparing a subject for the T-cell therapy, including the following:

Claims

1. A composition comprising anti-CD19 CAR-T cells for a method of treating cancer in a patient suitable for chimeric antigen receptor (CAR)-T cell therapy, wherein a preconditioning agent comprising cyclophosphamide is administered in combination with a preconditioning agent comprising fludarabine, and the CAR-T cells are administered to the patient on the day of administration (day 0) but prior to administration, if the serum levels of interleukin-15 ("IL-15"), interleukin-7 ("IL-7"), and at least one additional cytokine selected from the group consisting of monocyte chemotactic protein 1 ("MCP-1"), C-reactive protein ("CRP"), interferon-gamma-inducible protein 10 ("IP-10"), and any combination thereof are increased, and in the case of IL-15, the serum level is increased by at least 5 times.

2. The composition according to claim 1, wherein the cancer is lymphoma.

3. The composition according to claim 1, wherein the serum level of IL-15 increases by at least five times, the serum level of IL-7 increases by at least two times, the serum level of either MCP-1 or CRP increases by at least 1.5 times, and the serum level of IP-10 increases by at least two times, and the cancer is lymphoma or leukemia.

4. Approximately 500mg / m ２ / day ~ approx. 600mg / m ２ The patient was given a daily dose of cyclophosphamide for three days, and approximately 30 mg / m². ２ The composition according to claim 2, wherein a daily dose of fludarabine is administered daily for three days.

5. Approximately 900mg / m ２ / day ~ approx. 1000mg / m ２ The daily dose of cyclophosphamide is administered, and approximately 25 mg / m² is given. ２ The composition according to claim 3, wherein a daily dose of fludarabine is administered daily for three days.

6. The composition according to claim 4, wherein the cyclophosphamide and fludarabine are administered on the 5th, 4th, and 3rd day prior to the administration of the anti-CD19 CAR-T cells.

7. The composition according to claim 5, wherein the cyclophosphamide is administered on the -2 day prior to the administration of the CAR-T cells, and the fludarabine is administered on the -4 day, the -3 day, and the -2 day.

8. The composition according to any one of claims 1, 3, 5, and 7, wherein the cancer is leukemia.

9. The composition according to claim 8, wherein the leukemia is acute lymphoblastic leukemia (ALL).

10. A method for identifying a patient suitable for treatment of cancer with anti-CD19 CAR-T cell therapy, comprising identifying the patient as suitable for treatment of cancer with T cell therapy if, after administration of cyclophosphamide and fludarabine, on the day of administration of CAR-T cells (day 0) but prior to administration, serum levels of interleukin-15 ("IL-15"), interleukin-7 ("IL-7"), and at least one additional cytokine selected from the group consisting of monocyte chemotactic protein 1 ("MCP-1"), C-reactive protein ("CRP"), interferon-gamma-inducible protein 10 ("IP-10"), and any combination thereof are elevated.

11. The method according to claim 10, wherein the cancer is lymphoma.

12. The method according to claim 10, wherein the serum level of IL-15 increases by at least five times, the serum level of IL-7 increases by at least two times, the serum level of either MCP-1 or CRP increases by at least one.5 times, and the serum level of IP-10 increases by at least two times, and the cancer is lymphoma or leukemia.

13. About 500 mg / m ２ / day to about 600 mg / m ２ / day of cyclophosphamide is administered daily for 3 days, and about 30 mg / m ２ / day of fludarabine is administered daily for 3 days, the method according to claim 11.

14. Approximately 900mg / m ２ / day ~ approx. 1000mg / m ２ The daily dose of cyclophosphamide is administered, and approximately 25 mg / m² is given. ２ The method according to claim 12, wherein a daily dose of fludarabine is administered daily for three days.

15. The method according to claim 13, wherein the cyclophosphamide and fludarabine are administered on the 5th, 4th, and 3rd day prior to the administration of the CAR-T cells.

16. The method according to claim 14, wherein the cyclophosphamide is administered on the -2 day prior to the administration of the CAR-T cells, and the fludarabine is administered on the -4 day, the -3 day, and the -2 day.

17. The method according to any one of claims 10, 12, 14, and 16, wherein the cancer is leukemia.

18. The method according to claim 17, wherein the leukemia is acute lymphoblastic leukemia (ALL).

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