In CAR-T cell in situ recruitment, reprogramming, and release

Abstract

To provide a pharmaceutical composition for treating cancer. [Solution] The pharmaceutical composition comprises a hydrogel matrix containing a chimeric antigen receptor (CAR), an NK cell receptor, an NK T cell receptor, or a viral vector encoding a T cell receptor, wherein the hydrogel matrix is ​​formulated for administration to a target. Preferably, the pharmaceutical composition further comprises the hydrogel matrix containing one or more antibodies, cytokines, or costimulatory molecules that activate T cells, macrophages, natural killer (NK) cells, NK T cells, tumor-infiltrating NK cells (TINK), tumor-infiltrating lymphocytes (TIL), or medullary lymphocytes (MIL).

Description

[Background technology]

[0001] I. Background technology 1. T lymphocytes redirected to chimeric antigen receptors (CARs) (CAR-T cells) Adoptive cell transfer has produced impressive clinical effects against B-cell malignancies, and also in other Thai It is also effective against cancer. Current clinical-scale manufacturing methods for CAR-T cells This involves extensive ex vivo cell manipulation, namely isolation of autologous T cells, and CARs. Transduction using a viral vector, and pre-injection CAR-T for return to the patient. This requires the ex vivo proliferation of cells. The elaborate and time-intensive ex vivo procedure is essentially The costs are substantial, specifically the procedure takes 3-4 weeks and costs approximately $500,000. These costs limit the possibility of extending this technology to other cancers and patients. This involves mobilizing and modifying T cells to circumvent the challenges associated with existing CAR T cells. A new method is needed to achieve this. [Overview of the project]

[0002] II. Outline of the Invention 2. Disclosed are methods and compositions related to hydrogels containing chemiactive attractants.

[0003] 3. In one embodiment, as described herein, a hydrogermatic compound comprising one or more chemiactive substances is used. A disclosure is made in which one or more chemoattractants are CC motif chemokine ligands (CCL). )1(CCL1), CCL5, CCL19, CCL21, CCL22, CCL28, C- XC motif chemokine ligand (CXCL)1 (CXCL1), CXCL9, CXC L10, CXCL11, CXCL12, M-CSF, GM-CSF, MCP-1, MCP -3, CCL2, CCL3, CCL7, CCL20, CX3CL1, BRAK, IL-1 Includes 2, S1P, and / or MCP2.

[0004] 4. Also, chimeric antigen receptors (CARs), NK cell receptors (anti-CD3 antibodies, CD1d, Immunoglobulin Fc fragment or anti-i-Fcγ receptor (FcγRIII) antibody Including, but not limited to, NK T cell receptors and / or T cell receptors (e.g., For example, a viral vector encoding an antigen-specific T cell receptor (e.g., lenticular virus). Preceding viruses (including retroviruses, adenoviruses, or adeno-associated viruses) A hydrogel matrix of any form is disclosed herein.

[0005] 5. In one aspect, T cells, natural killer (NK) cells, NK T cells, tumors Infiltrating lymphocytes (TILs), medullary lymphocytes (MILs), tumor-infiltrating NK cells (TINKs), One or more antibodies, cytokines, and / or other antibodies that activate dendritic cells or macrophages. Alternatively, a hydrogel matrix of any of the prior embodiments further comprising a co-stimulatory molecule is described herein. This will be disclosed in writing. For example, antibodies include anti-CD3, anti-CD28, and anti-inducible costimulators (ICOS). ), it can contain anti-CD40L, anti-DAP10, and cytokines such as IL-2, I It may contain L-7, IL-15, IL-21, TNF-α, or IFN-γ.

[0006] 6. Furthermore, a hydrogel matrix of any of the prior embodiments further comprising a chemotherapeutic agent Disclosed herein.

[0007] 7. In one embodiment, the Specified Use of Bioresponsive Hydrogels of Any of the Preceding Embodiments is used in this Specified Use of Bioresponsive Hydrogels A method for treating, preventing, inhibiting, and / or reducing cancer or metastasis in a subject, including is disclosed. For example, a method for treating cancer of the present subject matter including administering to a subject a hydrogel matrix containing one or more chemoattractants is disclosed herein, and the one or more chemoattractants are C-C motif chemokine ligand (CCL) 1 (CCL1), C CL5, CCL19, CCL21, CCL22, CCL28, C-X-C motif chemokine ligand (CXCL) 1 (CXCL1), CXCL9, CXCL10, CXCL11 , CXCL12, M-CSF, GM-CSF, MCP-1, MCP-3, CCL2, C CL3, CCL7, CCL20, CX3CL1, BRAK, IL-12, S1P, and / or or MCP2, and the chemoattractants attract and retain immune cells (e.g., T cells, NK cells, NK T cells, TIL, MIL, TINK, dendritic cells and / or macrophages) in the hydrogel.

[0008] 8. Also disclosed herein is any method of the preceding aspects for treating, preventing, inhibiting, and / or reducing cancer or metastasis, wherein the matrix further comprises an immune checkpoint inhibitor and / or a chemotherapeutic agent.

[0009] 9. In one aspect, disclosed herein is any method of the preceding aspects for treating, preventing, inhibiting and / or reducing cancer or metastasis, wherein the hydrogel further comprises a chimeric antigen receptor (CAR), an NK cell receptor (including, but not limited to, an anti-CD3 antibody, CD1d, an immunoglobulin Fc fragment or an anti-Fcγ receptor (FcγRIII) antibody), an NK T cell receptor, and / or a T cell receptor (TCR) (antigen specificity ​ comprising a viral vector encoding (including T cell receptors). In one embodiment, the viral vector is transduced into immune cells, and the transduced immune cells are released from the hydrogel against cancer The viral vector can be introduced into the hydrogel matrix about 1 day to about 14 days before administration to the subject or after administration of the hydrogel matrix to the subject, which is understood and also intended herein. That is, the viral vector can be introduced into the hydrogel in vivo.

[0010] 10. Also disclosed herein is any method of the preceding embodiments for treating, preventing, inhibiting and / or reducing cancer or metastasis, and the immune cells are released about 1 week to about 1 2 weeks after administration of the hydrogel.

[0011] 11. In one embodiment, any method of the preceding embodiments for treating, preventing, inhibiting and / or reducing cancer or metastasis is disclosed herein, and one or more chemoattractants are released about 1 hour after administration of the hydrogel to about 12 weeks after administration of the hydrogel.

[0012] [[0]] [[ID=)[[1]]12. In one embodiment, a method of transducing immune cells of a subject (e.g., T cells, NK cells, NK T cells, TIL or MIL) is disclosed herein, the method comprising, for the subject, one or more chemoattractants (e.g., CCL1, CCL5, CCL19, CCL21, CCL 22, CCL28, CXCL1, CXCL9, CXCL10, CXCL11, CXCL1 2, M-CSF, GM-CSF, MCP-1, MCP-3, CCL2, CCL3, CCL 7, CCL20, CX3CL1, BRAK, IL-12, S1P, and / or MCP ​​​2) and introduced genes (e.g., CAR, NK cell receptor (anti-CD3 antibody, CD1d, immuno) It contains noglobulin Fc fragments or anti-Fcγ receptor (FcγRIII) antibodies, (not limited to these), NK T cell receptors, and / or TCRs (antigen-specific TCs) Viral vectors that encode (including, but not limited to, R) (e.g., lenticular) Hydrogen (including retroviruses, adenoviruses, or adeno-associated viruses) This includes administering the virus vector to the target before administration or to the target. Approximately 1 to 14 days after administration of the hydrogel matrix, the patient is guided to the hydrogel matrix. It is understood and intended that it may be possible to enter. That is, the virus vector The ter can be introduced into a hydrogel in vivo.

[0013] 13. Furthermore, any of the prior embodiments of a method for transduction into immune cells is disclosed herein. One or more chemoattractants were present approximately 1 hour after administration of the hydrogel to approximately 12 hours after administration of the hydrogel. It will be released weekly.

[0014] 14. In one embodiment, any of the methods of the prior embodiments for transduction into immune cells is described herein. Disclosed, hydrogels further include T cells, NK cells, NK T cells, TILs or MILs It includes one or more antibodies, cytokines, and / or costimulatory molecules that activate the following: , antibodies include anti-CD28, CD3, B7-1, B7-2, anti-inducible costimulatory factor (ICOS) ICOS ligand, anti-CD27, CD70, 4-1BBL, anti-41-BB, anti-CD40 L, CD40, anti-DAP10, anti-CD30, CD30L, anti-TIM-1, anti-TIM-2, Anti-TIM-3, anti-CD44, anti-NK1.1, lectin-like transcript-1 (LLT-1), anti-C D137, CD48, MICA, anti-2B4, and anti-glucocorticoid-induced tumor necrosis factor It can contain receptor-associated proteins (GITRs), and cytokines include IL-2 and IL It may contain IL-7, IL-15, IL-21, TNF-α, or IFN-γ. [Brief explanation of the drawing]

[0015] III. Brief Description of the Drawings 15. The attached drawings are incorporated into and constitute part of this specification, and some Embodiments are illustrated, and the compositions and methods of this disclosure are described together with the specification.

[0016] [Figure 1] 16. This illustrates the generation of CAR-T cells in situ. The transplanted scaffold releases chemokines, recruiting host T cells. The viral vector reprograms the T cells with tumor-specific CAR constructs, and the CAR-modified T cells migrate out and attack the cancer. [Figure 2] 17. This diagram illustrates the manufacturing process of a macroporous gel via cryogelization. [Figure 3] 18. SEM images of gels prepared by the freeze-drying regimen (-20°C) are shown. [Figure 4] 19. We demonstrate that CXCL10 regulates T cell recruitment to the transplanted scaffold. Figure 4A shows the experimental schedule, and Figure 4B shows the FACS analysis of CD3+ T cells recruited to a blank (control) or CXCL10-releasing scaffold. [Figure 5] 20. This demonstrates that pre-seeded and recruited T cells can be transduced within the transplanted scaffold. Experimental schedules (5A, 5C) and FACS analyses (5B, 5D) for GFP+ T cells pre-seeded in the scaffold or recruited to the scaffold by CXCL10. D shows further impact of CD3 / CD28 and IL-2 in the scaffold. [Figure 6] 21. Demonstrates in situ formation in CAR T cells. [Figure 7] 22. This shows in vivo transduction of T cells mobilized by the CD19 CAR virus. [Figure 8] 23. A schematic diagram of the creation of a mouse Burkitt lymphoma tumor model is shown. [Figure 9A] 24. Luminescence images of tumor size at 19, 24, 29, 33, and 43 days after tumor inoculation are shown for animals that were untreated, IV-administered with CD19 CAR T cells, or treated with a CCI-alginate scaffold (CCI-Alg). [Figure 9B] 25. Total flux and percentage change in body weight at various time points after tumor inoculation in animals that were untreated, IV-administered with CD19 CAR T cells, or treated with a CCI-alginate scaffold (CCI-Alg). [Figure 10] 26. This shows the number of CAR+ cells per 100 μl of blood from mice transplanted with a CCI scaffold or intravenously injected with CAR-T cells. Blood was collected from the cheek of the mice, red blood cells were hemolyzed, and cells were stained with Hu-CD45, Hu-CD3, and CAR.19 antibodies and analyzed by flow cytometry. [Modes for carrying out the invention]

[0017] IV. Modes for Carrying Out the Invention 27. Before the compound, composition, article, apparatus and / or method is disclosed and described Unless otherwise specified, these are based on specific synthesis methods or specific recombinant biotechnology. The Gee method is not limited to, or unless otherwise specified, it is not limited to, a specific reagent. Therefore, it should be understood that it is naturally subject to change. Also, the terms used in this specification The terms used are intended to describe only specific embodiments and are not intended to be limiting. I want you to understand that it's not just a thing.

[0018] A.Definition 28. In this specification and the appended claims, the following definitions are used: Let's refer to some of the terms.

[0019] 29. The singular "a", "an" as used in the specification and attached claims. " and "the" refer to multiple reference pairs unless the context clearly indicates otherwise. This includes elephants. Therefore, for example, a reference to "medicinal carrier" means two or more such carriers. Includes mixtures, etc.

[0020] 30. In this specification, the range is defined as a range from one specific value using "approximately" and / or The term "approximately" can be used to represent a range up to a specific other value. When such a range is expressed, Another embodiment includes a range from one specific value to / or the other specific value. By using the antecedent "approximately," when a value is expressed as an approximation, it indicates a specific It will be understood that different embodiments will result depending on the value of . Furthermore, each endpoint of the range Whether it is related to other endpoints or independent of other endpoints, it is significant. It will be understood that there are many values ​​disclosed herein, and each value is also In addition to the value itself, the specific value is also disclosed herein using "approximately". This is understood. For example, if the value "10" is disclosed, then "approximately 10" is also disclosed. In addition, if a certain value is disclosed in a manner that can be properly understood by those skilled in the art, The value "less than or equal to (less than or equal to)", the value "more than or equal to ( It is understood that the possible range between values ​​(greater than or equal to that value) is also disclosed. If the value "10" is disclosed, then "less than or equal to 10 (10 or less)" Not only that, but "more than or equal to 10 (10 or more)" is also disclosed. Through this, the data is provided in many different formats, and this data includes the endpoint and the starting point. It is also understood that it represents a range for any combination of data points. For example If a specific data point "10" and a specific data point 15 are disclosed, then from 10 and 15... Greater than, more than or equal to, less than, less than or In addition to being equal to (less than or equal to) and being equal to 10 and 15, the range between 10 and 15 is also taken into consideration. It is understood that they exist. It is also understood that each unit between two specific units is disclosed. For example, if “10 and 15” are disclosed, then 11, 12, 13, and 1 Number 4 has also been disclosed.

[0021] 31. "Administration" to the subject includes any route through which the drug is introduced or delivered to the subject. Administration methods include oral, topical, intravenous, subcutaneous, and transcutaneous. Absorption (transdermal), intramuscular, intra-articular, parenteral, intraarteriole, intradermal, ventricular, Intracranial, intraperitoneal, intralesional, intranasal, rectal, vaginal, via inhalation, transplanted reservoir (im via a planted reservoir; parenteral (e.g., subcutaneous, intravenous, intramuscular). Internal, intra-articular, intra-synovial bursa, intrasternal, intrathecal, intraperitoneal, intrahepatic, intralesional, and intracranial injections. This can be done by any preferred route, including injection techniques. The terms used are "combined administration," "combined administration," "simultaneous administration," or "administered at the same time." "Administered" means that the compounds are administered at the same time, or essentially immediately after each other. In the latter case, the two compounds produce the same result if the compounds were administered at the same time. The drug is administered at a time when the observed results are so close that they cannot be distinguished. "Systemic administration" means... For example, through entry points into the circulatory or lymphatic system, a wide area of ​​the target body (for example, The drug is introduced or delivered to the target via a route that introduces or delivers the drug to more than 50% of the body. It refers to reaching the target area. In contrast, "local administration" refers to directly targeting the area of ​​the administration site or the administration site. The drug is introduced or delivered to an adjacent area, and a therapeutically significant amount of the drug is not introduced systemically. This refers to introducing or delivering a drug to a target via a specific route. For example, local administration. The administered drug is easily detectable locally near the administration site, but not in the distal parts of the target body. It is undetectable or detectable in negligible amounts. Administration includes self-administration and administration to others. This includes administration by means of [specific method].

[0022] 32. "Biocompatibility" generally means that a substance is non-toxic to the recipient and does not weigh heavily on the target. Generally refers to materials that do not cause significant adverse effects, and any metabolites or degradation products thereof. vinegar.

[0023] 33. "~includes" means that the composition, method, etc., includes the listed elements, but does not include other elements. It is not intended to exclude anything. Used to define compositions and methods. In this case, "essentially derived from ~" means that it includes the listed elements, but in terms of their combination... This means eliminating any element that is essentially important. The composition, essentially consisting of the elements defined in the details, is subject to trace contamination from isolation and purification methods. The substance and pharmaceutically acceptable carriers, such as phosphate-buffered saline and preservatives, are removed. Do not remove. "~consists of" means that elements and the composition of the present invention are not added in trace amounts to other components. This means eliminating the substantial methods and processes required to provide. Embodiments defined by (Transition term) are within the scope of the present invention. be.

[0024] 34. A "control" is an alternative subject or sample used in an experiment for comparative purposes. The control group may be "positive" or "negative."

[0025] 35. "Controlled release" or "sustained release" refers to the desired pharmacokinetic profile in vivo. To achieve this, it refers to releasing the drug in a controlled manner from a predetermined dosage form. "Release" is one form of drug delivery that establishes the desired kinetics of drug release, and / or This refers to the ability to manipulate dosage forms.

[0026] 36. The "effective dose" of a drug refers to the amount of drug sufficient to provide the desired effect. The amount of drug that is "effective" depends on the age and general condition of the subject, and whether it is a specific drug or multiple drugs. Depending on the many factors, it will vary from subject to subject. Therefore, the quantified "effective amount" It is not always possible to identify ". However, in any case However, the appropriate "effective amount" may be determined by those skilled in the art using routine experiments. Furthermore, as used herein, unless otherwise specified, the “effective dose” of a drug is: It can refer to an amount that covers both the therapeutically effective dose and the preventively effective dose. The "effective dose" of medication required varies depending on factors such as the age, sex, and weight of the subject. In some cases, the administration regimen can be adjusted to provide the best possible therapeutic response. For example, the dose may be administered in several divided doses daily, or depending on the urgency of the treatment situation. The dosage may be reduced proportionally to achieve the desired result.

[0027] 1. "Increase" means that the amount of a sign, disease, composition, symptom, or activity becomes consequently greater. This can refer to a change in symptoms, signs, activity, or composition of a statistically significant magnitude. It can be an increase in an individual value, median, or mean. Therefore, an increase As long as it is statistically significant, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 , 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, It can be an increase of 90, 95, or 100%. An increase is a multiple of an increase. It can also refer to an increase of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15. , 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 , 500, 600, 700, 800, 900, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 This includes a doubling of the increase.

[0028] 2. "Decrease" refers to a smaller gene expression, protein expression, symptoms, disease, composition, or state. It can represent any change that results in a certain amount of state or activity. A substance can also be a compound containing a substance. When the genetic output of a gene product is less than the output of a gene product that does not contain the substance, It is understood that this reduces the gene output of a gene. Furthermore, this reduction can lead to, for example, the symptoms of a disorder. The symptom may be a change in that the condition is lower than previously observed. In a statistically significant amount, the median or average of any of the following: condition, symptoms, activity, or composition. It can be a decrease in the mean. Thus, as long as the decrease is statistically significant, the decrease is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 4 5, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% of a decrease. The decrease can also refer to a decrease as a multiple. For example , the decrease is 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 6 0, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 8 00, 900, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 -fold decrease.

[0029] 3. "Inhibit", "inhibiting", and "inhibition" mean reducing activity, response, condition, disease, or other biological parameters. This can include, but is not limited to, complete disappearance of activity, response, condition, or disease. This can also include, for example, a 10% reduction in activity, response, condition, or disease compared to the natural or control level. Thus, the reduction can be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount between these compared to the natural or control level. 4. As used herein, the terms "prevent", "preventing", "prevention", and their grammatical variations refer to the onset or recurrence of a disease and / or the associated

[0030] symptoms of the disease. symptoms of the disease. To partially or completely delay or prevent one or more symptoms of a disease, or for the subject to be affected by a disease. To acquire or prevent the reacquisition of a disease or symptoms associated with such a disease This refers to methods that reduce the risk associated with acquiring or reacquiring one or more types of status.

[0031] 5. A “medically acceptable” ingredient is one that is not biologically or otherwise undesirable. It can refer to a component that does not have a significant or undesirable biological effect. Without causing any adverse effects, or in any harmful manner with any other component of the preparation containing it. Without interaction, incorporated into the pharmaceutical formulation of the present invention as described herein. , may be administered to the target. When used in relation to administration to humans, this term is generally The ingredients meet the required standards for toxicity testing and manufacturing testing, or the ingredients are made from rice. This implies that the ingredient is listed in the guidelines for non-active ingredients created by the National Food and Drug Administration.

[0032] 6. A "medically acceptable carrier" (sometimes called a "carrier") is generally a safe carrier. Carriers or excipients useful for preparing whole and non-toxic pharmaceutical or therapeutic compositions. This refers to a drug that is permitted for veterinary and / or human pharmaceutical or therapeutic use. Including the body. The terms “carrier” or “medically acceptable carrier” refer to phosphate-buffered diet. Saltwater, water, emulsion (oil / water emulsion or water / oil emulsion, etc.) and / or may include, but is not limited to, various types of wetting agents used herein. In this context, the term "carrier" can refer to any excipient, diluent, filler, salt, buffer, or stabilizer. , well known in the art for use as a solubilizer, lipid, stabilizer, or in pharmaceutical formulations This includes, but is not limited to, other materials as further described herein.

[0033] 7. Like a "pharmacologically active" derivative or analog, "pharmacologically active" (and (Simply put, "active") means having the same type of pharmacological activity as the parent compound, and to approximately the same degree. derivatives or analogs (e.g., salts, esters, amides, conjugates, metabolites, etc.) It can refer to a substance, fragment, etc.

[0034] 8. "Therapeutic drug" refers to any composition that has beneficial biological effects. The effects include therapeutic effects, such as the treatment of disorders or other undesirable physiological conditions, and And preventive effects, for example, disorders or other undesirable physiological conditions (e.g., non-immunogenic conditions) This includes both prevention of (n). These terms also include salts, esters, amides, and precursors. This includes proagents, active metabolites, isomers, fragments, analogs, etc. The pharmaceutically acceptable This also includes pharmacologically active derivatives. When the term "therapeutic drug" is used, or specifically When a specific drug is identified, the term can refer to the drug itself, as well as to other medically permitted drugs. Accepted, pharmacologically active salts, esters, amides, precursors, conjugates, active substitutes Please understand that this also includes metaproducts, isomers, fragments, and analogues.

[0035] 9. The “therapeutic effective amount” or “therapeutic effective dose” of a composition (for example, a composition containing a drug) This refers to the amount that is effective in achieving the desired treatment outcome. In some embodiments, The desired treatment outcome is the suppression of type 1 diabetes. In some embodiments, the desired treatment outcome is The treatment involves suppressing obesity. The therapeutically effective dose of a given drug is typically used for the treatment of a disorder or disease. The type and severity of the disease, as well as factors such as the age, sex, and weight of the subject, may vary. It will become so. This term is also effective in promoting desired therapeutic effects such as pain relief. It can also refer to the amount of medication or the rate of medication delivery (e.g., the amount over time). The desired therapeutic effect depends on the condition to be treated, the resistance of the target, the drug to be administered, and / or This refers to pharmaceutical formulations (for example, the efficacy of a therapeutic agent, the concentration of the drug in the formulation, etc.), as well as by persons skilled in the art. It will vary depending on various other factors that are understood. In some cases, the desired biological A therapeutic or medical reaction may occur over a period of several days, weeks, or years depending on the dosage of the composition. This is achieved after administration to the target.

[0036] 10. In this specification and the appended claims, the following definitions are used: Let's refer to some of the terms.

[0037] 11. “Optional” or “optionally” means the event or situation described thereafter. The situation may or may not occur, and this description is based on the possibility that the event or situation may occur. This means that it includes cases where it occurs and cases where it does not.

[0038] 12. Throughout this application, various publications are referenced. To more fully describe the level, the disclosures in these publications are, by reference, their entirety. This is incorporated herein. The disclosed references also include the text on which those references rely. The materials discussed within are explained individually and specifically by reference. It will be included in the detailed specifications.

[0039] B. Composition 13. In addition to the components used to prepare the disclosed compositions, other components disclosed herein The compositions used in the method are also disclosed. These and other materials are disclosed herein. This is disclosed, and combinations, subsets, interactions, and groups of these materials are also disclosed. If these compounds are present, then each of the various individual and collective combinations and rearrangements Specific references to may not be explicitly disclosed, but each is specifically mentioned herein. It is understood that this is intended and described. For example, certain hydrates containing chemical attractants A hydrogel matrix containing a chemoattractant is disclosed and discussed. When considering the numerous modifications that can be made to numerous molecules including this one, it is not possible to make a specific decision. If not, a hydrogel matrix containing a chemiactive attractant and its possible modifications Every conceivable combination and rearrangement of these can be specifically envisioned. Therefore, the molecule In addition to classes A, B, and C, classes D, E, and F of molecules are also disclosed and combined. When AD, an example of a molecule, is disclosed, and each is not described individually Even so, each is a combination of meanings intended individually and collectively, AE, AF, BD, BE, BF, CD, CE, and CF are deemed to be disclosed. Similarly, any subset or combination of these is also disclosed. For example, the subgroups AE, BF, and CE are considered to be disclosed. This concept includes steps in a method for producing and using the disclosed composition. This applies to all aspects of this application, but is not limited to these. Therefore, feasible If various additional steps exist, each of these additional steps is optional of the disclosed method. It is understood that this can be accomplished by a specific embodiment or combination of embodiments.

[0040] 14. Chimeric antigen receptor (CAR) immunotherapy uses the patient's own T cells and NK cells. CAR-T cells, CAR-NK cells, and CAR-T cells, respectively, of macrophages. - Including gene modification of NK-T cells or CAR macrophages (CARMA) These are types of cancer immunotherapy. These CAR cells can target specific tumor antigens. It expresses antibody-based chimeric antigen receptors. This process involves autologous T cells, Isolation of NK cells, NK T cells, or macrophages, and CAR-encoding viruses Transduction by a chemist, and CAR-T cells, CAR-NK cells, CAR-NK-T cells This includes ex vivo proliferation of CARMA and subsequent reinfusion into the patient. Despite clinical success, clinical-grade CAR immune cells ( For example, the process of generating CAR T cells is cumbersome, time-consuming, and costly. The procedure still involves implementing CAR immunotherapy as the standard treatment for cancer. It remains a major barrier to generating CAR immune cells. Approaches that can reduce time and cost are clinically very important, and CAR T The effectiveness of cytoplasm, CAR-NK cells, CAR-NK-T cells and / or CARMA therapy It can be dramatically improved.

[0041] 15. Generating tumor-specific CAR immune cells in situ is equivalent to CAR-T cells. This can solve many of the challenges associated with ex vivo generation of chimeric antigen receptors (C). AR) refers to immune cells against tumor-associated antigens (e.g., CAR T cells, CAR-NK cells). It is a synthetic receptor that redirects the specificity of CAR-NK-T cells or CARMA cells. CD19 injection. CAR T cells showed significant improvement in clinical trials with CD19. It has shown antitumor effects. CAR-T therapy was approved by the FDA in 2017. T cell harvesting, ex vivo activation, gene modification and cell proliferation, and subsequent disease The injection into the patient takes time (about 4 weeks) and is also expensive (about $500,000). Assuming that this already places a significant burden on the healthcare system, millions of people Providing CAR immunotherapy to cancer patients is a very significant challenge. The detailed description states that T cells are loaded with factors that recruit T cells and viral vectors that encode CARs. In a macroporous scaffold, CAR immune cells can be generated in situ. This shows that CAR immune cells (for example) may have clinical significance. Then, CAR T cells, CAR-NK cells, CAR-NK-T cells (or CARMA) are produced. By reducing the time and costs associated with achieving this, the effectiveness of CAR immunotherapy can be dramatically improved. It can be improved to that extent.

[0042] 16. This specification describes the use of bioactive macroporous scaffolds to induce CAR immunotherapy in patients. A strategy for generating cells in situ is disclosed. The scaffold mobilizes immune cells. They then activate and reprogram these cells into CAR immune cells, and introduce CAR immune cells into the body. The cells are released and directed towards the tumor (Figure 1). The release of chemokines from the scaffold is controlled by T cells. (including, but not limited to, TIL and MIL), NK cells (including TINK, (Not limited to these), using NK T cells, dendritic cells and / or macrophages as a scaffold. It forms an attractive gradient. The macroporous scaffold is mediated by chemokines, which stimulate T cells and NK cells. Promotes the infiltration and embedding of cells, NK T cells, dendritic cells, and / or macrophages. Along with cytokines and antibodies, T cells, NK cells, NK T cells, dendritic cells and / Alternatively, it promotes the survival and proliferation of macrophages, and also facilitates transduction by viral particles. It provides an interface. After the chemokines are completely released, their depletion is a scaffolding. Migration of CAR-T cells, CAR-NK cells, CAR-NK-T cells, or CARMA cells This approach promotes NK cells, NK T cells, dendritic cells and / or ma Clophages are recruited to a biomaterial scaffold and then injected into the CAR gene by a viral vector. Therefore, it is reprogrammed and released into the bloodstream from the scaffold, resulting in T cells in situ. Cells, CAR-T cells, CAR-NK cells, CAR-NK-T cells, or CARMA fragments This leads to the generation without [unclear]. Therefore, in one embodiment, as specified herein, one or more chemical attractants A hydrogel matrix containing a substance is disclosed.

[0043] 17. The purpose of chemoattractants is to attract immune cells (e.g., T cells, natural killer (NK) cells). Cells, NK T cells, tumor-infiltrating lymphocytes (TILs), medullary lymphocytes (MILs), tumor infiltration Lubricated NK cells (TINK), dendritic cells, and / or macrophages are attracted to the hydrogel. It is understood that this is what is intended to be done, and is intended in the present invention. Examples of one or more chemoattractants that may be present include, but are not limited to, CC motif chemocacti. In ligand (CCL)1 (CCL1), CCL5, CCL19, CCL21, CCL2 2. CCL28, CXC motif chemokine ligand (CXCL)1 (CXCL1) , CXCL9, CXCL10, CXCL11, CXCL12, M-CSF, GM-CSF , MCP-1, MCP-3, CCL2, CCL3, CCL7, CCL20, CX3CL1 Examples include BRAK, IL-12, S1P, and / or MCP2.

[0044] 18. The hydrogels disclosed herein may use any preferred biodegradable polymer. It can be manufactured using [a certain method]. "Polymer" refers to a relatively high molecular weight polymer, either natural or synthetic. Poly Non-specific examples of polymers include polyethylene, rubber, and cellulose. Synthetic polymers - is usually formed by the addition polymerization or condensation polymerization of monomers. The term refers to a polymer formed from two or more different repeating units (monomer residues). Refers to. Not as a limitation but as an example, copolymers include alternating copolymers and random copolymers. - may be a block copolymer or a graft copolymer. In certain embodiments, it may also be a block copolymer. The various block segments of a block copolymer may themselves contain copolymers. The term "polymer" is used to refer to natural polymers, synthetic polymers, and homopolymers. This includes, but is not limited to, heteropolymers or copolymers, addition polymers, etc. It encompasses all forms of polymers.

[0045] 19. In one embodiment, the hydrogel is a biocompatible polymer (e.g., alginate). Such polymers may also gradually introduce CAR T cells into the tissue, CAR NK cells, CAR NK T cells, CARMA, TIL, MIL, TINK and / or It can also play a role in releasing MIL. When used herein, biocompatibility The polymers are not limited to, but include polysaccharides such as alginates, chitosan, and hyaluronic acid. , hydrophilic polypeptides, proteins such as collagen, fibrin and gelatin, poly -L-glutamic acid (PGS), gamma-polyglutamic acid, poly-L-aspartic acid Poly(amino acids) such as poly-L-serine or poly-L-lysine, polyethylene glycerides Polypropylene glycol (PEG), polypropylene glycol (PPG), and poly(ethylene oxide) Polyalkylene glycols and polyalkylene oxides such as (PEO), poly(O (Xyethylated polyol), poly(olefin alcohol), polyvinylpyrrolidone), Poly(hydroxyalkyl methacrylamide), poly(hydroxyalkyl methacrylamide) T), poly(saccharide), poly(hydroxy acid), poly(vinyl alcohol), poly( Polyhydro(lactic acid), poly(glycolic acid), and poly(lactic acid-coglycolic acid) are examples of polyhydro(lactic acid-coglycolic acid). Poly(xic acid, poly-3 hydroxybutyrate or poly-4 hydroxybutyrate, etc.) Hydroxyalkanolates, polycaprolactones, poly(orthoesters), polyanhydrides Poly(phosphazene), poly(lactide-cocaprolactone), tyrosine polycarbonate Polycarbonates such as nates, polyamides (including synthetic and natural polyamides), poly Peptides and poly(amino acids), polyesteramides, polyesters, poly(dioxa) Non-polyester, poly(alkylene alkylate), hydrophobic polyether, polyurethane, polyester Polyester, polyacetal, polycyanoacrylate, polyacrylate, polymer Chil methacrylate, polysiloxane, poly(oxyethylene) / poly(oxypropyl) (n) Copolymer, polyketal, polyphosphate. Polyhydroxyvalerate, polya Lukilen oxalate, polyalkylene succinate, poly(maleic acid), and this These copolymers are examples. Biocompatible polymers include polyamides and polycarbonates. Polyphosphate, polyalkylene, polyalkylene glycol, polyalkylene oxide, poly Alkylene terephthalate, polyvinyl alcohol (PVA), methacrylate PVA ( m-PVA), polyvinyl ether, polyvinyl ester, polyvinyl halide, Livinylpyrrolidone, polyglycolide, polysiloxane, polyurethane and these Polymers, alkylcellulose, hydroxyalkylcellulose, cellulose ether, Cellulose esters, nitrocellulose, acrylic esters and methacrylic esters Polymers, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylcellulose, hydroxypropylcellulose Roxypropyl methylcellulose, hydroxybutyl methylcellulose, cellulose acetate cellulose propionate, cellulose acetate butyrate, cellulose acetate f Talat, carboxyethylcellulose, cellulose triacetate, sodium cellulose sulfate Poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate) Poly(isobutyl methacrylate), poly(hexyl methacrylate) Poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl Poly(methyl acrylate), poly(isopropyl acrylate) Poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene Polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethylene Poly(vinyl alcohol), poly(vinyl acetate, polyvinyl acetate) Loridopolystyrene and polyvinylprirolidone, their derivatives, and their linear and This also includes branched copolymers and their block copolymers, as well as blends thereof. It is possible to do so. Examples of biodegradable polymers include polyester and poly(ortho-s). Poly(ethyleneamine), Poly(caprolactone), Poly(hydroxybutyrate) Poly(hydroxyvalerate), polyanhydride, poly(acrylic acid), polyglycol Poly(urethane), polycarbonate, polyphosphate ester, polyphosphoric acid, These derivatives, their linear and branched copolymers, and their block copolymers These include ma, as well as blends thereof.

[0046] 20. In some embodiments, the particles are poly(lactic acid), poly(glycolic acid), and biocompatible and / or biodegradable poly(lactic acid-coglycolic acid) Contains polyester or polyanhydrous. The particles are the following polyesters, i.e., Honmei The glycolic acid unit referred to as "PGA" in the detailed document, and poly-L-lactic acid, poly-D-lactic acid, and Li-D,L-lactic acid, poly-L-lactide, poly-D-lactide, and "PLA" are also mentioned. In detailed texts, lactic acid units such as poly-D,L-lactide 5 and poly(e-cap) Caprolactone units, such as loractone, which are collectively referred to herein as "PCL," and A homopolymer containing [the specified compound], and characterized by a lactic acid:glycolic acid ratio, and referred to as "PLGA" and Honmyo Poly(lactic acid-coglycolic acid) and poly(lactide-coglycolic acid) are collectively referred to in the detailed document. Copolymers containing lactic acid and glycolic acid units, such as various forms of licorice; It may also contain polyacrylate and one of its derivatives. Examples of polymers include PLGA-PEG or PLA-PEG copolymers. Polyethylene glycerides, which are collectively referred to herein as "pegylated polymers," in various forms. Copolymers of pol (PEG) and the aforementioned polyesters are also examples. In this state, the PEG region is covalently bonded to the polymer and cleavable by a linker, resulting in a "PEG" region. A polymer can be produced. In one embodiment, the polymer is at least 60 , 65, 70, 75, 80, 85, 89, 90, 91, 92, 93, 94, 95, 96, Contains 97, 98, or 99 percent acetal pendant groups.

[0047] 21. The triblock copolymers disclosed herein include, for example, polyethylene glyco Polyvinyl alcohol (PEG), polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone (PV) P), polyethylene oxide (PEO), poly(vinylpyrrolidone-co-vinyl acetate) (t) Polymethacrylate, polyoxyethylene alkyl ether, polyoxyethylene Castor oil, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic acid-glycolic acid) ) Acid, poly(lactic acid co-glycolic acid) (PLGA), hydroxymethylcellulose, hydr It contains core polymers such as cellulose derivatives like roxypropylcellulose.

[0048] 22. In one embodiment, the hydrogel matrix further activates and / or maintains the immune system. Antibodies, chemokines, and cytokines for maintenance (e.g., CD28, CD3, IL-2) It can include IL-7, IL-15, IL-21, IFN-γ, and TNF-α. ru.

[0049] 23. The disclosed hydrogel matrix contains one or more immune checkpoint inhibitors It is understood that and / or chemotherapeutic agents may further be included, and also specified herein It is intended to be used in the disclosed hydrogel matrix. This may include any chemotherapy agent, but is not limited to the following: Abemaciclib, Abiletorone acetate, Abitrexat e(Methotrexate), Abraxane(Paclitaxel albumin) (Stabilized nanoparticle formulations), ABVD, ABVE, ABVE-PC, AC, AC-T, Adc etris (Brentuximab Vedotin), ADE, Ado-Trast uzumab Emtansine, Adriamycin (Doxorubicin salt) (Salt), Afatinib dimaleate, Afinitor (Everolim (us), Akynzeo (Netupitant and Palonosetron hydrochloride) ), Aldara (Imiquimod), Aldesleukin, Alecensa (Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed 2 sodium), Aliqopa (Copanlisib hydrochloride) ), Alkeran for injection (Melphalan hydrochloride), Alkeran tablets (Mel phalan), Aloxi (Palonosetron hydrochloride), Alunbrig( Brigatinib), Ambochlorin (Chlorambucil), Am boclorin Chlorambucil), Amifostine, aminolebri nic acid, Anastrozole, Aprepitant, Aredia (Pamidro nate 2 sodium), Arimidex (Anastrozole), Aromas in (Exemestane), Arranon (Nelarabine), Arsen ic Trioxide, Arzerra (Ofatumumab), Erwinia chrysanthemi asparaginase, Atezolizumab, Avasti n (Bevacizumab), Avelumab, Axitinib, Azaciti dine, Bavencio (Avelumab), BEACOPP, Becenum( Carmustine), Beleodaq (Belinostat), Belinos tat, Bendamustine hydrochloride, BEP, Besponsa (Inotuzu mab Ozogamicin), Bevacizumab, Bexarotene, B exxar (Tositumomab and Iodine I 131 Tositum omab), Bicalutamide, BiCNU (Carmustine), Ble omycin, Blinatumomab, Blincyto (Blinatumoma b), Bortezomib, Bosulif (Bosutinib), Bosutin ib, Brentuximab Vedotin, Brigatinib, BuMel, Busulfan, Busulfex (Busulfan), Cabazitaxel, Cabometyx (Cabozantinib-S-テツト) Cabozant inib-S-CAF, Campath(Alemtuzumab), Ca mptosar (Irinotecan drug) Capecitabine CAP OX, Carac(Fluorouracil-alcoholic), Carboplatin, C.S ARBOPLATIN-TAXOL、Carfilzomib、Carmubris(C armustine)、Carmustine、Carmustine Implant Casodex(Bicalutamide), CEM, Ceritinib, Cer ubidine(Daunorubicin solvent) Cervarix(drug HP V Discount Cetuximab, CEV, Chlorambucil, CHLO RAMBUCIL-PREDNISONE、CHOP、Cisplatin、Cladr ibine、Clafen(Cyclophosphamide)、Clofarabi and Clofarex(Clofarabine)、Clolar(Clofarab ine)、CMF、Cobimetinib、Cometriq(Cabozantin ib-S-マート) Copanlisib National Park COPDAC COPP CO PP-ABV、Cosmegen(Dactinomycin)、Cotellic(C obimetinib) Crizotinib CVP Cyclophospham ide, Cyphos (Ifosfamide), Cyramza (Ramuciruma (b) Cytarabine, Cytarabine, Cytosar-U( Cytarabine), Cytoxan(Cyclophosphamide), Da brafenib, Dacarbazine, Dacogen (Decitabine) , Dactinomycin, Daratumumab, Darzalex (Darat umumab), Dasatinib, Daunorubicin hydrochloride, Daunor Ubicin hydrochloride and Cytarabine liposomes, Decitabine, D Defibrotide sodium, Defitelio(Defibrotide sodium) um), Degarelix, Denileukin Diftitox, Denosu mab, DepoCyt (Cytarabine liposome), Dexamethaso ne, Dexrazoxane hydrochloride, Dinutuximab, Docetaxel, Doxil (Doxorubicin hydrochloride liposome), Doxorubicin hydrochloride Salt, Doxorubicin hydrochloride liposome, Dox-SL (Doxorubicin Liposome hydrochloride, DTIC-Dome (Dacarbazine), Durvalu mab, Efudex (Fluorouracil - topical), Elitek (Rasb uricase, Ellence (Epirubicin hydrochloride), Elotuzum ab, Eloxatin (Oxaliplatin), Eltrombopag Ola mine, Emend(Aprepitant), Empliciti(Elotuzu mab), Enasidenib mesylate, Enzalutamide, Epirub Icin hydrochloride, EPOCH, Erbitux (Cetuximab), Eribuli n-mesylate, Erivedge (Vismodegib), Erlotinib hydrochloride Erwinaze (Erwinia chrysanthemi asparaginase), Ethyol (Amifostine), Etopophos (Etoposide P phosphate), Etoposide, Etoposide Phosphate, Evacet (Doxorubicin hydrochloride liposome), Everolimus, E vista, (Raloxifene hydrochloride), Evomela (Melphalan salt acid salt), Exemestane, 5-FU (for Fluorouracil injection), 5-F U (Fluorouracil - topical use), Fareston (Toremifene) , Farydak (Panobinostat), Faslodex (Fulvestr ant), FEC, Femara (Letrozole), Filgrastim, Fl udara (Fludarabine phosphate), Fludarabine Phosphate sphate, Fluoroplex (Fluorouracil - topical use), Fluo Fluorouracil Injection, Fluorouracil - Topical Use, Flu amide, Folex(Methotrexate), Folex PFS(Meth otrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FO LFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Foloty n(Pralatrexate), FU-LV, Fulvestrant, Gardas il (recombinant HPV quadrivalent vaccine), Gardasil 9 (recombinant HPV nonavalent vaccine) ), Gazyva (Obinutuzumab), Gefitinib, Gemcita bine hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABIN E-OXALIPLATIN, Gemtuzumab Ozogamicin, Gemz ar (Gemcitabine hydrochloride), Gilotrif (Afatinib dimaleate) Gleevec (Imatinib mesylate), Gliadel (Carmine) stine implant), Gliadel wafer (Carmustine implant) Lant, Glucarpidase, Goserelin acetate, Halave n(Eribulin mesylate), Hemangeol(Propranolol hydrochloride) Salt), Herceptin (Trastuzumab), HPV bivalent vaccine, recombinant, HPV nonavalent vaccine, recombinant; HPV tetravalent vaccine, recombinant; Hycamtin (Top otecan hydrochloride), Hydrea (Hydroxyurea), Hydroxyur ea, Hyper-CVAD, Ibrance (Palbociclib), Ibrit umomab Tiuxetan, Ibrutinib, ICE, Iclusig (Po Natinib hydrochloride, Idamycin (Idarubicin hydrochloride), Idar Ubicin hydrochloride, Idelalisib, Idhifa (Enasidenib) Rate), Ifex (Ifosfamide), Ifosfamide, Ifosfam idum (Ifosfamide), IL-2 (Aldesleukin), Imati nib mesylate, Imbruvica (Ibrutinib), Imfinzi (Du rvalumab), Imiquimod, Imlygic (Talimogene L aherparepvec), Inlyta(Axitinib), Inotuzuma b Ozogamicin, Interferon Alfa-2b, recombinant, inter Leukin-2 (Aldesleukin), Intron A (recombinant interferon α- 2b) Iodine I 131 Tositumomab and Tositumom ab, Ipilimumab, Iressa (Gefitinib), Irinotec AN hydrochloride, Irinotecan hydrochloride liposome, Istodax (Romidep sin), Ixabepilone, Ixazomib Citrate, Ixempr a(Ixabepilone), Jakafi(Ruxolitinib phosphate) , JEB, Jevtana (Cabazitaxel), Kadcyla (Ado-Tr astuzumab emtansine), Keoxifene (Raloxifen) e hydrochloride), Kepivance (Palifermin), Keytruda (Pem brolizumab), Kisqali (Ribociclib), Kymriah ( Tisagenlecleucel), Kyprolis (Carfilzomib), Lanreotide acetate, Lapatinib Ditosylate, La rtruvo (Olaratumab), Lenalidomide, Lenvatin ib mesylate, Lenvima (Lenvatinib mesylate), Letrozo le, Leucovorin Calcium, Leukeran(Chlorambu cil), Leuprolide acetate, Leustatin (Cladribi ne), Levulan (Aminolevulinic Acid), Linfoli zin (Chlorambucil), LipoDox (Doxorubicin hydrochloride) Liposomes, Lomustine, Lonsurf (Trifluridine an d Tipiracil hydrochloride, Lupron (Leuprolide acetate) Lupron Depot (Leuprolide acetate), Lupron D epot-Ped (Leuprolide acetate), Lynparza (Olap arib), Marqibo (Vincristine Sulfate liposome), M atulane (Procarbazine hydrochloride), Mechlorethamine Hydrochloride, Megastrol acetate, Mekinist (Trametinib) Melphalan, Melphalan hydrochloride, Meraptopurine, M esna, Mesnex (Mesna), Methazolastone (Temozo) lomide), Methotrexate, Methotrexate LPF(Me thotrexate), Methylnaltrexone Bromide, Mex ate(Methotrexate), Mexate-AQ(Methotrexate) ), Midostaurin, Mitomycin C, Mitoxantrone hydrochloride, M itozytrex (mitomycin C), MOPP, Mozobil (Plerixa for), Mustargen (Mechlorethamine hydrochloride), Mutaam ycin (mitomycin C), Myleran (Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogam) icin), Nanoparticle Paclitaxel (Paclitaxel Albumin-stabilized nanoparticle formulation), Navelbine (Vinorelbine saturates) (Cy ester), Necitumumab, Nelarabine, Neosar (Cy clophosphamide), Neratinib Maleate, Nerlyn x (Neratinib maleate), Netupitant and Palonoset ron hydrochloride, Neulasta (Pegfilgrastim), Neupogen ( Filgrastim), Nexavar (Sorafenib Tosylate), Nilandron (Nilutamide), Nilotinib, Nilutami de, Ninlaro (Ixazomib Citrate), Niraparib Toshi Rate monohydrate, Nivolumab, Nolvadex (Tamoxifen Citrate) ), Nplate (Romiplostim), Obinutuzumab, Odom zo(Sonidegib), OEPA, Ofatumumab, OFF, Olapar ib, Olaratumab, Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase), Ondansetron hydrochloride, On Ivyde (Irinotecan hydrochloride liposome), Ontak (Denileuk in Diftitox), Opdivo (Nivolumab), OPPA, Osim ertinib, Oxaliplatin, Paclitaxel, Paclitaxe l-albumin-stabilized nanoparticle formulations, PAD, Palbociclib, Palifrm in, Palonosetron hydrochloride, Palonosetron hydrochloride and Net upitant, Pamidronate 2 sodium, Panitumumab, Pa nobinostat, Paraplat(Carboplatin), Parapla tin (Carboplatin), Pazopanib hydrochloride, PCV, PEB, Pe gaspargase, Pegfilgrastim, Peginterferon A lfa-2b, PEG-intron (Peginterferon Alfa-2b), Pembrolizumab, Pemetrexed Disodium, Perjet a(Pertuzumab), Pertuzumab, Platinol(Cispla tin), Platinol-AQ (Cisplatin), Plerixafor, P omalidomide, Pomalyst (Pomalidomide), Ponat inib hydrochloride, Portrazza (Necitumumab), Pralatrex ate, Prednisone, Procarbazine hydrochloride, Proleukin (Aldesleukin), Prolia (Denosumab), Promacta (Eltrombopag Olamine), Propranolol hydrochloride, Pro venge(Sipuleucel-T), Purinethol(Mercaptop urine), Purixan (Mercaptopurine), Radium 22 3 Dichloride, Raloxifene hydrochloride, Ramucirumab, R asburicase, R-CHOP, R-CVP, recombinant human papillomavirus (H) PV) bivalent vaccine, recombinant human papillomavirus (HPV) nonavalent vaccine, recombinant human Topapillomavirus (HPV) quadrivalent vaccine, recombinant interferon α-2b, Re gorafenib, Relistor (Methylnaltrexone bromide), R -EPOCH, Revlimid (Lenalidomide), Rheumatrex (Methotrexate), Ribociclib, R-ICE, Rituxan( Rituximab), Rituxan Hycela (Rituximab and human) Hyaluronidase), Rituximab, Rituximab and human hyaluronidase -ze, Rolapitant hydrochloride, Romidepsin, Romiplostim, Rubidomycin (Daunorubicin hydrochloride), Rubraca (Ruc aparib camsylate), Rucaparib camsylate, Ruxolitini β-phosphate, Rydapt (Midostaurin), Sclerosol intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T, Soma Tuline Depot (Lanreotide acetate), Sonidegib Sorafenib tosylate, Sprycel (Dasatinib), STANF ORD V, Sterile Talc Powder (Talc), Steritalc(Talc), Stiv arga (Regorafenib), Sunitinib maleate, Sutent ( Sunitinib (maleate), Sylatron (Peginterferon A) lfa-2b), Sylvant (Siltuximab), Synribo (Omac etaxine (mepesuccinate), Tabloid (Thioguanine), T AC, Tafinlar (Dabrafenib), Tagrisso (Osimert inib), Talc, Talimogene Laherparepvec, Tamo xifen Citrate, Tarabine PFS (Cytarabine), Tarc eva (Erlotinib Hydrochloride), Targretin (Bexarotene), Tasigna (Nilotinib), Taxol (Paclitaxel), Tax otere (Docetaxel), Tecentriq, (Atezolizumab ), Temodar (Temozolomide), Temozolomide, Tem sirolimus, Thalidomide, Thalomid (Thalidomi de), Thioguanine, Thiotepa, Tisagenlecleuce l, Tolak (Fluorouracil - for topical use), Topotecan Hydrochloride, T oremifene, Torisel (Temsirolimus), Tositumo mab and Iodine 131 Tositumomab, Totect (Dexrazox ane Hydrochloride), TPF, Trabectedin, Trametinib, Trast uzumab, Treanda (Bendamustine Hydrochloride), Trifluri dine and Tipiracil Hydrochloride, Trisenox (Arsenic Trioxide), Tyke rb (Lapatinib Ditosylate), Unituxin (Dinutuximab ), Uridine Triacetate, VAC, Vandetanib, VAMP , Varubi (Rolapitant Hydrochloride), Vectibix (Panitumu mab), VeIP, Velban (Vinblastine Sulfate), Ve lcade (Bortezomib), Velsar (Vinblastine sulfate) Vemurafenib, Venclexta (Venetoclax), Vemurafenib netoclax, Verzenio (Abemaciclib), Viadur (Le uprolide acetate, Vidaza (Azacitidine), Vinb lastine sulfate, Vincasar PFS (Vincristine sulfate Vincristine sulfate, Vincristine sulfate Tryposomes, Vinorelbine tartrate, VIP, Vismodegib , Vistogard (Uridine triacetic acid), Voraxaze (glucarpidase) ), Vorinostat, Votrient (Pazopanib hydrochloride), Vyxe os (Daunorubicin hydrochloride and Cytarabine liposomes), We llcovorin (Leucovorin calcium), Xalkori (Crizo tinib), Xeloda (Capecitabine), XELIRI, XELOX , Xgeva (Denosumab), Xofigo (Radium 223 Dich loride), Xtandi (Enzalutamide), Yervoy (Ipil) imumab), Yondelis (Trabectedin), Zaltrap (Zi v-Aflibercept), Zarxio (Filgrastim), Zejula (Niraparib tosylate monohydrate), Zelboraf (Vemurafeni b), Zevalin (Ibritumomab Tiuxetan), Zinecar d(Dexrazoxane hydrochloride), Ziv-Aflibercept, Zofran (Ondansetron hydrochloride), Zoladex (Goserelin acetate) ), Zoledronic Acid, Zolinza (Vorinostat), Zo meta(Zoledronic Acid), Zydelig(Idelalisib ), Zykadia (Ceritinib), and / or Zytiga (Avilatero) Examples include (Abiraterone acetate). Checkpoint inhibitors and However, it is not limited to PD-1 (Nivolumab (BMS-93) 6558 or MDX1106), CT-011, MK-3475), PD-L1 (MD X-1105(BMS-936559), MPDL3280A, MSB0010718C ), PD-L2(rHIgM12B7), CTLA-4(ipilimumab(Ipilimu mab)(MDX-010), Tremelimumab (CP-6) 75,206)), IDO, B7-H3(MGA271), B7-H4, TIM3, LA An example is an antibody that blocks G-3 (BMS-986016).

[0050] 24. In one aspect, also disclosed in this specification are T cells, natural killer (N) cells. K) cells, NK T cells, dendritic cells, macrophages, tumor-infiltrating NK cells (TINK), One or more that activate tumor-infiltrating lymphocytes (TILs) or medullary lymphocytes (MILs) Further comprising antibodies, cytokines and / or costimulatory molecules as disclosed herein. A hydrogel matrix is ​​disclosed. For example, the antibody is anti-CD28, CD3, B7- 1, B7-2, anti-inducible costimulatory factor (ICOS), ICOS ligand, anti-CD27, CD 70, 4-1BBL, anti-41-BB, anti-CD40L, CD40, anti-DAP10, anti-CD3 0, CD30L, anti-TIM-1, anti-TIM-2, anti-TIM-3, anti-CD44, anti-NK1. 1. Lectin-like transcript-1 (LLT-1), anti-CD137, CD48, MICA, anti-2B 4 and anti-glucocorticoid-induced tumor necrosis factor receptor-associated protein (GITR) It is possible to do this, and cytokines include IL-2, IL-7, IL-15, IL-21, and TNF. It may contain -α or IFN-γ.

[0051] 25. One advantage of the disclosed hydrogel is that after immune cells come into contact with the hydrogel, Immune cells, for example, chimeric antigen receptors (e.g., antibodies or s that target cancer-specific antigens). cFv), T cell receptor (e.g., antigen-specific T cell receptor), NK cell receptor (anti-CD3 Antibodies, CD1d, immunoglobulin Fc fragments, or anti-Fcγ receptors (FcγRI) II) Includes, but is not limited to, antibodies, and includes transgenes such as NK T cell receptors. This means that transduction can be easily performed. The T antigen is a substance produced by tumor cells. It is a protein that induces an immune response (especially an immune response mediated by T cells). The antigen-binding domain may be a natural ligand for an antibody or T antigen, or an MHC molecule. It may be a molecule that recognizes peptides derived from the T antigen presented by. The choice of binding domain is thought to depend on the specific type of cancer being treated. T antigens are well known in the art, for example, glioma-associated antigens and carcinoembryonic antigens. (CEA), EGFRvIII, IL-llRa, IL-13Ra, EGFR, FAP, B7H3, Kit, CA LX, CS-1, MUC1, BCMA, bcr-abl, HE R2, β-human chorionic gonadotropin, alpha-fetoprotein (AFP), ALK, CD19, CD123, cyclin BL, lectin-reactive AFP, Fos-related antigen 1, A DRB3, thyroglobulin, EphA2, RAGE-1, RUL, RU2, SSX2, A KAP-4, LCK, OY-TESl, PAX5, SART3, CLL-1, Fucosil G M1, GloboH, MN-CA IX, EPCAM, EVT6-AML, TGS5, Totelomerase reverse transcriptase, polysialic acid, PLAC1, RUL RU2(AS), intestinal carboxylesterase, lewisY, sLe, LY6K, m ut hsp70-2, M-CSF, MYCN, RhoC, TRP-2, CYPIBI, BORIS, prostase, prostate-specific antigen (PSA), PAX3 , PAP, NY-ESO-1, LAGE-la, LMP2, NCAM, p53, p53 alteration Allomorph, Ra mutant, gp100, prostain, OR51E2, P ANX3, PSMA, PSCA, Her2 / neu, hTERT, HMWMAA, HAV CR1, VEGFR2, PDGFR-beta, survivin and te Lomerase, Legmain, HPV E6, E7, Sperm Protein 1 7. SSEA-4, tyrosinase, TARP, WT1, prostate cancer tumor antigen-1 (PCT) A-1), ML-IAP, MAGE, MAGE-A1, MAD-CT-1, MAD-CT -2, MelanA / MART 1, XAGE1, ELF2M, ERG(TMPRSS2 ETS fusion gene), NA17, neutrophil elastase, sarcoma translocation break ic point, NY-BR-1, ephnnB2, CD20, CD22, CD24, CD 30, CD33, CD38, CD44v6, CD97, CD171, CD179a, anti drogen receptor, FAP, insulin growth factor (IGF)-I, IGFII, IGF-I receptor, GD2, o-acetyl-GD2, GD3, GM3, GPRC5D, GPR20, CXORF61, folate receptor (FRa), folate receptor β, ROR1, Flt3, TAG7 2, TN Ag, Tie 2, TEM1, TEM7R, CLDN6, TSHR, UPK2 and mesothelin.

[0052] 1. Delivery of the composition to cells 26. Transduction of immune cells can be performed by any means known in the art. In one aspect, transduction of immune cells can be performed via a viral vector encoding a transgene (e.g., CAR). Thus, in one aspect, what is disclosed herein is a viral vector encoding a transgene (e.g., chimeric antigen receptor (CAR), T cell receptor , NK cell receptor and / or NK T cell receptor), for example, a lentivirus, retrovirus, adenovirus or adeno-associated virus) further included in any of the disclosed hydrogels.

[0053] 27. There are many compositions and methods that can be used for delivery of nucleic acids to cells in vitro or in vivo. These methods and compositions can be mainly classified into two classes, namely virus-based delivery systems and non-virus-based delivery systems. For example, nucleic acids are transported by many direct transport systems, such as electroporation and lipofacial injection. Action, calcium phosphate precipitate, plasmid, viral vector, viral nucleic acid, formula Transmitted via nucleic acids, phages, cosmids, etc., or via cells or carriers, such as cations. It can be delivered via the transport of genetic material by sex liposomes, etc. Suitable means for transfection such as a chemical transfectant or Physical and mechanical methods such as electroporation and direct diffusion of DNA are, for example, Wolff,JAet al.Science 247,1465-1468(19 90), and Wolff, JANature, 352, 815-818 (1991 ) is described. Such methods are well known in the art and are not described herein. It can be easily applied to the composition and method described. In particular cases, the method is It is thought that these modifications are made to function particularly well in large DNA molecules. Furthermore, these The method targets specific diseases and cell populations by using the targeted features of the carrier. It can be used for chemical purposes.

[0054] (1) Retroviral vector 28. Retroviruses are all types, subfamilies, and genera of the Retroviridae family. Retroviruses are animal viruses belonging to the virus family, which includes Tropism. Vectors are generally referred to as Verma, IM, or Retroviral vectors. It is described in the section for gene transfer.

[0055] 29. Retroviruses are essentially packages containing nucleic acid cargo. Nucleic acid cargo carries a packaging signal, which in turn allows replicated daughter molecules to form. This can be efficiently packaged within the package coat. In addition to the package signal In replication within the system, and in packaging the replicated virus, There are many key molecules. Typically, retroviral genomes are protein coats. It includes the gag, pol, and env genes involved in the production of [unclear]. Typically, gag, p The ol and env genes are replaced with foreign DNA, which is then transported to target cells. Retroviral vectors are typically packaged for incorporation into the package coat. The sequence that sends the aging signal and the start signal for the gag transcription unit, and the sequence required for reverse transcription The ment (containing a primer binding site for binding reverse transcription tRNA primers) and DNA During synthesis, terminal repeat sequences induce switching of the RNA strand, and the synthesis of the second strand of DNA synthesis... A purine-rich 5'→3'LTR that functions as a priming site, and an insertion site in the host genome. A specific arrangement near the end of the LTR allows for the insertion of retroviruses in their DNA state. The column includes, and by removing the gag, pol and env genes, approximately 8kb of the outer column is removed. The sequence is inserted into the viral genome, reverse transcribed, replicated, and becomes a novel retroviral particle. This can be packaged. The amount of nucleic acid depends on the size of each transcript, 1 It is sufficient for the delivery of multiple genes. The insert contains positive and It is preferable that the product has a negative selection marker.

[0056] 30. In most retroviral vectors, the replication mechanism and packaging tamper Because the cleavage molecules (gag, pol, and env) have been removed, the vector is typically... They can be generated by introducing them into a cell line for packaging. Pack aging cell lines are cell lines that are trans fected or transformed by retroviruses containing replication and packaging mechanisms, but lack any packaging signals. When a vector carrying the selected DNA is transfected into these cell lines, the vector containing the target gene is replicated and packaged into new retroviral particles by the mechanism provided in cis by helper cells. The genomes related to the mechanism are not packaged because they lack the signals they need.

[0057] (2) Adenovirus vector 31. The construction of replication-deficient adenoviruses is described in Berkner et al., J.V irology 61:1213-1220(1987), Massie et al. , Mol.Cell.Biol.6:2872-2883(1986), Haj-Ahm ad et al., J.Virology 57:267-274(1986), Da vidson et al., J.Virology 61:1226-1239(19 87), Zhang”Generation and identification of recombinant adenovirus by liposome-me diated transfection and PCR analysis”Bio Techniques 15:868-872(1993). The advantage of using these viruses as vectors is that the range in which they can spread to other types of cell types is limited, which means that the cells they initially infected [[ID=�6]] ​​​​​​ Although it can only replicate within itself, it is unable to form novel infectious virus particles. Recombinant adenoviruses affect the airway epithelium, hepatocytes, vascular endothelium, CNS parenchymal tissue, and many others. It is possible to perform highly efficient gene transfer after direct in vivo delivery to other tissue sites. As shown (Morsy, J. Clin. Invest. 92:1580-1586( 1993), Kirshenbaum (J.Clin). Invest.92:381- 387(1993), Roessler, J.Clin.Invest.92:1085 -1092(1993), Moullier, Nature Genetics 4:1 54-159(1993), La Salle, Science 259:988-99 0 (1993), Gomez-Foix, J. Biol. Chem. 267:25129 -25134(1992), Rich, Human Gene Therapy 4:4 61-476(1993), Zabner, Nature Genetics 6:75 -83(1994), Guzman,Circulation Research 73 :1201-1207(1993), Bout, Human Gene Therapy 5:3-10(1994), Zabner,Cell 75:207-216(199 3), Caillaud, Eur. J. Neuroscience 5:1287-12 91 (1993), and Ragot, J. Gen. Virology 74:501- 507 (1993). Recombinant adenoviruses bind to specific cell surface receptors, and Afterward, the virus, through receptor-mediated endocytosis, either wild-type or non-renewable. Similar to adenoviruses, gene transfer is performed by internalization. (Chardonnet and Dales, Virology 40:462-477) (1970), Brown and Burlingham, J. Virology 12: 386-396 (1973), Svensson and Persson, J. Virol ogy 55:442-449(1985), Seth et al., J. Virol .51:650-655(1984), Seth et al.,Mol.Cell.B iol.4:1528-1533(1984), Varga et al., J. Vir 65:6061-6070 (1991), Wickham et al. Cell 73:309-319(1993).

[0058] 32. The viral vector is based on adenovirus from which the E1 gene has been removed. Furthermore, these virions are generated in cell lines such as the human 293 cell line. In a preferred embodiment, both E1 and E3 genes are removed from the adenovirus genome. It can be done.

[0059] (3) Adeno-associated virus vector 33. Another type of viral vector is based on adeno-associated virus (AAV). This imperfect parvovirus can infect many types of cells. Furthermore, because it is non-pathogenic to humans, it is a desirable vector. AAV-type vector It can transport approximately 4-5kb, and wild-type AAV has chromosome 19 (e.g., AA It is known that it can be stably inserted into the V implantation site 1 (AAVS1). A vector having suitable embedding characteristics is preferred. A particularly preferred implementation of this type of vector. The form is P4.1, manufactured by Avigen, San Francisco, CA. It is a C vector, which is the thymidine kinase gene of herpes simplex virus HSV-t k, and / or a marker gene (e.g., encoding green fluorescent protein (GFP)). It can include genes.

[0060] 34. In other types of AAV viruses, AAV is ligated to heterologous genes in an operational manner. Located on both sides of at least one cassette containing a promoter that induces cell-specific expression. It includes a pair of reverse terminal repeats (ITRs). In this context, heterogeneous means AAV or This refers to a nucleotide sequence or gene that does not naturally exist in B19 parvovirus. vinegar.

[0061] 35. Typically, the AAV and B19 coding regions are removed, resulting in a safer environment. This has resulted in a cytotoxic vector. AAV ITR or its modifications are It possesses infectivity and site-specific integration capabilities, but is not cytotoxic, and its promoter is fine. It induces cell-specific expression. U.S. Patent No. 6,261,834 relates to the AAV vector. The materials used are incorporated herein by reference.

[0062] 36. The vectors disclosed herein can be incorporated into mammalian chromosomes without substantial toxicity. Provides DNA molecules.

[0063] 37. Genes inserted into viruses and retroviruses are usually the target gene product. Includes promoters and / or enhancers useful for expression control. Promoters generally A DNA sequence that functions when it is located in a relatively fixed position relative to the transcription start site. The promoter is necessary for the fundamental interaction between RNA polymerase and transcription factors. It includes a core element that is used, and also has elements and response elements upstream of it. It may include [something].

[0064] (4) Viral vectors for large payloads 38. Molecular genetic experiments using large human herpesviruses revealed that large heterologous DN Cloning and disseminating the A fragment, which is susceptible to infection by the herpesvirus. A method that can be established in cells has been provided (Sun et al., Natu re Genetics 8:33-41, 1994, Cotter and Robert Son, Curr Opin Mol Ther 5:633-644, 1999). child These large DNA viruses (herpes simplex virus (HSV) and Epstein-B) The virus (EBV) implants >150kb of human heterologous DNA fragments into specific cells. It has the ability to deliver [something]. Recombinant EBV, as episomal DNA, enters infected B cells. It can be maintained as a large DNA fragment. Individual clones can be up to 330kb. The togenome-derived inserts were transported and were considered genetically stable. Maintenance of these episomes In this case, a specific EBV nucleoprotein (EBNA1) is constitutively present during EBV infection. It is necessary for the protein to be expressed. Furthermore, these vectors allow for the large amount of protein to be expressed. It can be used in transfection to produce transients in Toro. Yes, it is possible. Herpesvirus amplicon systems also use DNA fragments >220kb. To package and infect cells in which DNA can be stably maintained as episomes It is being used.

[0065] 39. Other useful systems include, for example, replicated and host-limited non-replicating vaccines. Near-viral vectors are one example.

[0066] 2. Expression System 40. Nucleic acids delivered to cells typically contain expression regulatory systems. For example, viruses Genes inserted into retrovirus systems are usually useful for regulating the expression of the target gene product. Includes promoters and / or enhancers. Promoters generally relate to the transcription start site. It is a DNA sequence that functions when it is located in a relatively fixed position. The promoter is The core required for the fundamental interaction between RNA polymerase and transcription factors It includes an element, and may also include an element and a response element upstream of it.

[0067] a) Virus promoters and enhancers 41. Preferred promoters that control transcription from vectors in mammalian host cells These may originate from various sources (for example, the genome of a virus), for example For example, Polyoma, Simian virus 40 (SV40), adenovirus, retrovirus, Hepatitis B virus, and most preferably cytomegalovirus, or heterozoan mammalian virus. The derived promoter (for example, one derived from the beta-actin promoter) may also be S The early and late promoters of the V40 virus also include the SV40 virus replication origin. It can be easily obtained as a 40 restriction enzyme-treated fragment (Fiers et al., Na ture, 273:113 (1978). Immediate early promo of human cytomegalovirus. The data can be easily obtained as a HindIII E-restricted processing fragment (Greenw ay, PJet al., Gene 18:355-360 (1982). Of course. Hmm, promoters derived from host cells or closely related species are also useful in the present invention.

[0068] 42. An enhancer generally refers to a DNA sequence that functions at an indeterminate distance from the transcription start site. And that is the 5' side of the transcription unit (Laimins, L. et al., Proc. Natl Acad.Sci.78:993(1981)) or 3' side (Lusky, ML) In either .et al.,Mol.Cell Bio.3:1108(1983)) It is possible. Furthermore, enhancers can also reside within introns (Banerji ,J Let al., Cell 33:729(1983)), and coding distribution It can also exist inside the column itself (Osborne, TF et al., Mol Cell Bio. 4:1293 (1984). 4: They are usually 10-300 bp. They are the leaders, and they function in cis. Enhancers are nearby promo It plays a role in enhancing transcription that occurs from the source. Enhancers also mediate transcriptional regulation. It often contains response elements. Promoters are also response elements that mediate transcriptional regulation. It may include enhancers, which often determine the regulation of gene expression. In mammals Genes (globin, elastase, albumin, fetoprotein, and insulin) Many enhancer sequences derived from are now known, but typically, for general expression, Enhancers derived from eukaryotic virus are used. A preferred example is the replication origin of SV40. The enhancer of the later part (100-270 bp) of cytomegalovirus, early promoter Enhancers, late-side enhancers for the replication origin of polyomas, and adenovirus enhancers It is Nsa.

[0069] 43. Promoters or enhancers may have optical or specific properties that initiate their function. It can be specifically activated by a chemical event. The system is, for example, tetracycline It can be adjusted with reagents such as dexamethasone. Irradiation (e.g., gamma ray irradiation) Alternatively, exposure to alkylating chemotherapeutic agents can enhance gene expression in viral vectors. There is also a method that involves doing this.

[0070] 44. Configurations for maximizing the expression of the region of the transcription unit to be transcribed in a particular embodiment. As a promoter and / or enhancer, promoter and / or enhancer territory The area can be affected. In certain structures, promoters and / or enhancers The region is active in all types of eukaryotic cells, however, it is active at specific timings. It is expressed only in specific types of cells. A preferred promoter of this type is, The CMV promoter (650 bases) is preferred. Another preferred promoter is the SV40 promoter. In addition, cytomegalovirus (full-length promoter) and retroviral vector LTR be.

[0071] 45. Clone all specific regulatory elements and use them in specific types of cells (e.g.) It can be used to construct expression vectors that are selectively expressed in melanoma cells, for example. What can be done has been demonstrated so far. Glial fiber acidic protein (GFAP) promotion T is used to selectively express genes in glial-derived cells.

[0072] 46. ​​In eukaryotic host cells (yeast, fungi, insects, equipment, animals, humans, or nucleated cells) The expression vector used is also necessary for transcription termination, which can affect mRNA expression. These regions may contain sequences. These regions are the untranslated parts of mRNA encoding tissue factor proteins. It is transcribed as a polyadenylated segment at 3'. The 3' untranslated region is also the end of transcription. It includes the binding site. The transcription unit preferably also includes the polyadenylated region. One of these regions The benefit is that it increases the likelihood that the transcribed units will be processed and transported like mRNA. That is to say. The identification and use of polyadenylation signals in expression constructs is sufficient. It is established. In the gene construct to be introduced, the same type of polyadenylation signal is used. It is preferable to use. In a particular transcription unit, the polyadenylated region is the initial port of SV40. It originates from a riadenylation signal and consists of approximately 400 bases. Furthermore, the transcribed unit is different from other units. It contains the standard sequence, which, either alone or in combination with the above sequence, expresses from the construct. It is also preferable to improve or stabilize it.

[0073] b) Marker 47. Viral vectors may contain nucleic acid sequences that encode marker products. This marker product determines whether a gene has been delivered to a cell and subsequently expressed. It is used for this purpose. The preferred marker gene is E, which encodes β-galactosidase. This refers to the lacZ gene of .Coli and the green fluorescent protein.

[0074] 48. In some embodiments, the marker may be a selectable marker. Examples of suitable selectable markers for physical cells include dihydrofolate reductase (DHFR), thymid. Neomycin kinase, neomycin, neomycin analog G418, hydromycin and Pewter It is romycin. Mammalian host cells are successfully transformed using such a selectable marker. When subjected to selective pressure, transformed mammalian host cells survive. This is possible. There are two different categories that are widely used as selective regimens. Category 1 cells are those that lack viability when considered independently of cell metabolism and supplemental media. This is based on the use of mutant cell lines. Two examples include CHO DHFR- cells and These are mouse LTK cells. These cells contain nutrients such as thymidine or hypoxanthine. Without the addition of nutrients, they lack the ability to proliferate. These cells have a complete nucleotide compound. Because it lacks certain genes necessary for the synthesis pathway, the deficient nucleotides are present in the supplement medium. They cannot survive unless provided with a complete medium. As an alternative to supplementing the medium, By introducing the DHFR or TK gene into cells lacking the respective gene, their proliferation can be promoted. The requirements are altered. Transformation is performed by the DHFR or TK gene. The individual cells that were obtained are not viable in the non-supplemented medium.

[0075] 49. The second category of selection methods is dominant selection, which is of any type This is called a selection scheme, which is also used in cells, and does not require the use of mutant cell lines. The scheme typically involves using drugs that suppress the proliferation of host cells. Cells that have offspring carry proteins that induce drug resistance and survive selection. An example of dominant selection is the drug neomycin (Southern P. and Berg). ,PJ,Molec.Appl.Genet.1:327(1982)), Mikofe Nolic acid (Mulligan, RC and Berg, P., Science 209) :1422(1980)) or Hygromycin (Sugden, B. et al. This is the use of Mol.Cell.Biol.5:410-413(1985). 3 An example is the use of bacterial genes under eukaryotic regulation, with each being the appropriate drug, G41. 8 or neomycin (geneticin), xgpt (mycophenolic acid), or hi It induces resistance to glomycin. Another example is the neomycin analog G418. One example is puramycin.

[0076] 3. Delivery of pharmaceutical carriers / pharmaceutical products 50. As described above, the composition may also be administered in vivo in a pharmaceutically acceptable carrier. Yes, it is possible. "Medically acceptable" means that it is not undesirable from a biological or other standpoint. This means that the material does not cause any undesirable biological effects. Without rubbing, or in any harmful manner, it interacts with any other component of the pharmaceutical composition containing it. It can be administered to a subject along with nucleic acids or vectors without acting. The carrier is easily understood by those skilled in the art. As is well known, it is natural to minimize any breakdown of the active ingredients and target the target. It will be selected to minimize any harmful side effects.

[0077] 51. The composition may be administered orally, parenterally (e.g., intravenously), intramuscularly, or intraperitoneally. It can be administered by injection, transdermal administration, or extracorporeal administration, and by local intranasal administration or inhalation. This includes administration. As used herein, "local intranasal administration" means administration through one or both nostrils. This refers to the delivery of the composition to the nose and nasal cavity through a spray mechanism or a droplet mechanism. This may include delivery via aerosolization of nucleic acids or vectors. Inhalant The administration of the composition is via delivery through the nose or mouth via a spray or droplet mechanism. It can be delivered directly to any area of ​​the respiratory system (e.g., the lungs) via intubation. It is also possible. The exact amount of the required composition depends on the species, age, weight and overall condition of the subject, and the treatment. The severity of the allergic disorder being treated, the specific nucleic acid or vector used, and the form of administration. Depending on the subject, etc., it will vary. Therefore, the exact amount for all compositions is It is impossible to identify the appropriate amount. However, the appropriate amount can be determined with the teachings provided herein. This can be determined by those skilled in the art using only conventional experiments.

[0078] 52. Parenteral administration of the composition, when used, is generally done by injection. Its characteristics include: The injectable drug is available as a liquid solution or suspension, and the suspension is dissolved in the liquid before injection. It can be prepared in a conventional form, either as a solid or emulsion, suitable for the solution. It is possible. The relatively recently revised approach for parenteral administration maintains a constant dose. The use of a sustained-release or continuous-release system is used as such. For example, refer to the Specified See U.S. Patent No. 3,610,795 incorporated therein.

[0079] 53. The material is incorporated into a solution, suspension (e.g., microparticles, liposomes, or cells). They may be in the state of (being). These are transmitted via antibodies, receptors, or receptor ligands. It may target specific cell types. The following references describe how specific proteins target tumor tissue. This is an example of the use of this technology (Senter, et al., Bioconjugate). Chem.,2:447-451,(1991);Bagshawe,KD,Br .J.Cancer,60:275-281,(1989);Bagshawe,et al.,Br.J.Cancer,58:700-703,(1988);Senter ,et al.,Bioconjugate Chem.,4:3-9,(1993); Battelli,et al.,Cancer Immunol.Immunothe r.,35:421-425,(1992);Pietersz and McKenz i.e., Immunolog.Reviews, 129:57-80, (1992); oyo and Roffler, et al.Biochem.Pharmacol,42:2062 -2065, (1991). Vehicles, such as "Stealth" and antibodies are conjugates. Other liposomes (including lipid-mediated drug targeting of colon cancer), cell-specific liposomes Gandr-mediated receptor-mediated targeting of DNA, lymphocyte-induced tumor targeting, and in vivo Highly specific therapeutic retroviral targeting of mouse glioma cells. The following references are specific. This is an example of the use of this technology, in which the protein targets tumor tissue (Hughes et al. ., Cancer Research, 49:6214-6220, (1989); approximately and Litzinger and Huang, Biochimica et Bioph. ysica Acta, 1104:179-187, (1992). Generally, receptors are It is involved in the endocytosis pathway, either constitutively or ligand-induced. These receptors accumulate in clathrin-coated pits, and clathrin-coated They enter the cell via vesicles, pass through acidified endosomes, and are then sorted by receptors. It is then recycled to the cell surface, stored inside the cell, or lithosaturated. It is either broken down in the body or internalized. The internalization pathway is the uptake of nutrients. Furthermore, it removes activated proteins, clears macromolecules, and opportunistically invades viruses and toxins. It performs various functions such as entry, ligand dissociation and degradation, and regulation at the receptor level. Many receptors depend on the cell type, receptor concentration, ligand type, ligand valency, And depending on the ligand concentration, it follows one or more intracellular pathways. Receptor-mediated endoscopy The molecular and cellular mechanisms of itosis are outlined (Brown and Greene, D NA and Cell Biology 10:6,399-409(1991)).

[0080] a) Medically acceptable carriers 54. Compositions containing antibodies are used therapeutically in combination with pharmaceutically acceptable carriers. It is possible.

[0081] 55. Suitable carriers and formulations of said carriers are described in Remington: The Science e and Practice of Pharmacy (19th ed.)ed. ARGennaro,Mack Publishing Company,East This is explained in on, PA 1995. Typically, to make the formulation isotonic An appropriate amount of pharmaceutically acceptable salt is used in the formulation. Examples of pharmaceutically acceptable carriers Examples include physiological saline, Ringer's solution, and dextrose solution, but these Not limited to, the pH of the solution is preferably about 5 to about 8, more preferably about 7 to about 7.5. As a further carrier, a sustained-release formulation, for example, a solid hydrophobic polymer containing an antibody, is used. - A semipermeable matrix is ​​one example, and this matrix is ​​used in molded products, for example, films. It is in the form of liposomes or microparticles. A carrier, for example, administers the composition to be administered. It will be obvious to those skilled in the art that, depending on the path and concentration, there may be more preferable options.

[0082] 56. Pharmaceutical carriers are known to those skilled in the art. These are most typically sterile water, physiological food. Solutions for drug administration to humans, including saline solutions and solutions buffered at physiological pH. This would be a standard carrier. The composition can be administered intramuscularly or subcutaneously. Other The compound will be administered according to standard procedures used by those skilled in the art.

[0083] 57. The pharmaceutical composition, in addition to the selected molecule, contains a carrier, thickener, diluent, buffer, and preservative. It may contain agents and surfactants, etc. Pharmaceutical compositions include antibacterial agents, anti-inflammatory agents, anesthetics, etc. It may contain more than one species of active ingredient.

[0084] 58. Whether or not local or systemic treatment is desired for the pharmaceutical composition, and whether or not treatment is desired Depending on the area to be treated, it can be administered in several ways. Administration can be local (eye drops, vaginal, or direct injection). It can be administered intravenously (including nasally), orally, by inhalation, or parenterally, for example, intravenously, subcutaneously, or intraperitoneally. It can be administered by intravenous or intramuscular injection. The disclosed antibody can be administered intravenously or intraperitoneally. It can be administered intramuscularly, subcutaneously, intracavitarially, or transdermally.

[0085] 59. Preparations for parenteral administration include sterile aqueous solutions or non-aqueous solutions, suspensions and Examples of non-aqueous solvents include emulsions. Examples of non-aqueous solvents include propylene glycol and polyethylene. Glycols, vegetable oils such as olive oil, and injectable organic compounds such as ethyl oleate. It is a sterling. The aqueous carrier contains physiological saline and a buffering medium, and is water, alcoholic. Examples include aqueous solutions, emulsions, or suspensions. As a parenteral vehicle, sodium chloride Rium solution, dextrose Ringer's solution, dextrose and sodium chloride, lactate Examples include Ngel solution or non-volatile oils. As an intravenous vehicle, it provides fluids and nutrients. Examples include pleurinary infusions and electrolyte replacement solutions (such as those based on dextrose-ringer's solution). It contains preservatives and other additives, such as antimicrobial agents, antioxidants, chelating agents and inert gases. Other variations such as "S" may also exist.

[0086] 60. Formulations for topical administration include ointments, lotions, creams, gels, drops, and suppositories. Examples include agents, sprays, liquids, and powders. Conventional medicinal carriers are aqueous, powder, or oily. A base, thickener, etc. may be necessary or desirable.

[0087] 61. Compositions for oral administration: powder or granules, suspension in water or a non-aqueous medium. Examples include liquids or solutions, capsules, sachets or tablets. Thickeners, flavorings, diluents, Emulsifiers, dispersants, or binders may be desirable.

[0088] 62. Some of the compositions are hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid. and inorganic acids such as phosphoric acid, as well as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, Organic acids such as pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid By the reaction of, or with sodium hydroxide, aluminum hydroxide, potassium hydroxide, etc. Inorganic bases, as well as mono, di, trialkylamines and arylamines and substituted A pharmaceutically acceptable acid formed by the reaction with organic bases such as ethanolamine. Alternatively, it may be administered as a base addition salt.

[0089] b) Therapeutic use 63. The effective dose and schedule for administering the composition may be determined experimentally. Making such a decision falls within the scope of the technology in that field. The dosage range is a sufficiently large range of doses to produce the desired effect in which the symptoms of the disorder are affected. The dosage may cause undesirable side effects such as cross-reactions and anaphylactic reactions. It should not be excessively large. Generally, the dosage should be determined based on the patient's age, condition, sex, and disease. It varies depending on the degree, route of administration, and whether or not other drugs are included in the regimen. The dosage can be determined by the individual physician. In case of any contraindications, the dosage will be determined by the individual physician. It can be adjusted. The dosage can be changed, once a day or for several days. It can be administered in the above dosage. For appropriate dosages of a given class of pharmaceutical products: Guides can be found in the literature. For example, guides for selecting the appropriate antibody dosage. For example, literature on the therapeutic uses of antibodies, such as the Handbook of Monoclo nal Antibodies, Ferrone et al., eds., Noges Publications, Park Ridge, NJ, (1985) ch.2 2 and pp.303-357;Smith et al., Antibodies in Human Diagnosis and Therapy, Haber et al. al., eds., Raven Press, New York (1977)pp.3 It can be found in 65-389. The typical daily dose of the antibody used alone is above Depending on the factors listed, the daily dose may range from approximately 1 μg / kg to a maximum of 100 mg / kg (body weight) or... It could be in a range exceeding this.

[0090] C. Methods for transducing immune cells 64. The disclosed hydrogel contains immune cells (e.g., T cells, NK cells, NK T cells, dendritic cells, macrophages, TINK, TIL and / or MIL) It can be used to induce in vivo transduction of immune cells. This is understood and intended in this specification. In one aspect, what is disclosed herein is , target immune cells (e.g., T cells, NK cells, NK T cells, dendritic cells, macrophages) This is a method of transducing (J, TINK, TIL, or MIL) to a subject. , one or more chemiactive substances (e.g., CCL1, CCL5, CCL19, CCL21, C CL22, CCL28, CXCL1, CXCL9, CXCL10, CXCL11, CXC L12, M-CSF, GM-CSF, MCP-1, MCP-3, CCL2, CCL3, C CL7, CCL20, CX3CL1, BRAK, IL-12, S1P, and / or M CP2) and transgenes (e.g., CAR, NK cell receptor (anti-CD3 antibody, CD1d, Contains immunoglobulin Fc fragments or anti-Fcγ receptor (FcγRIII) antibodies However, it is not limited to this, but also includes T cell receptors (e.g., antigen-specific T cell receptors) or Viral vectors encoding NK T cell receptors (e.g., lentiviruses, retroviruses) Administer a hydrogel containing (virus, adenovirus or adeno-associated virus) This includes.

[0091] 65. Viral vectors should be administered to the target or hydrogermated before administration to the target. After administration of the drug, it can be introduced into the hydrogel matrix approximately 1 to 14 days later. It is understood, and intended, that the viral vector is It can be introduced into a hydrogel in vivo. For example, a viral vector can be introduced into a hydrogel. During gel formation, after gel formation, but before administering the hydrogel to the subject, or to the subject After administering the hydrogel, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, It can be implemented on the 14th, 15th, 16th, 17th, 18th, 19th, 20th, or 21st. ru.

[0092] 66. The disclosed chemiactive attractant leaches or is released into the hydrogel over time. It is understood, and intended, that hydrogels attract immune cells. The immune cells in question (e.g., T cells, NK cells, NK T cells, dendritic cells, macrophages, TINK (TIL or MIL) is a chimeric antigen receptor, NK T cell receptor, and NK cell Receptors (anti-CD3 antibody, CD1d, immunoglobulin Fc fragment or anti-Fcγ) This includes, but is not limited to, antibodies against the receptor (FcγRIII) or T cell receptor (anti- Transduction by transgenes such as (including, but not limited to, protospecific T cell receptors). This is possible. In one embodiment, the release of the chemiactive substance occurs from 1 hour after administration of the hydrogel. This can be approximately 12 weeks after administration of the hydrogel. For example, the release of chemotactic substances is After administering the hydrogel, approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 28, 30, 36, 42, 48, 60, 72 hours, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 28, 30, 31, 5, 6, 7 , 9, 10, 11 or 12 weeks. Therefore, in one embodiment, Disclosed in the specification is a method for transducing immune cells, comprising one or more chemoattractants. These substances are released approximately 1 hour after administration of the hydrogel to approximately 12 weeks after administration of the hydrogel. The release time depends on whether the chemical attractant is added to the hydrogel polymer before or after gelation. This can be adjusted by adding the chemical attractant after gelation. It provides a rapid release time, and its addition to the polymer during or before gelation is effective. It results in a delayed release.

[0093] 67. In one embodiment, immune cells are maintained by further stimulation, or It is recognized that the benefit of having those immune cells activated for treatment can be obtained. Therefore, in one embodiment, what is disclosed herein is a method of introducing traits, Hydrogels are used in cells, NK cells, NK T cells, dendritic cells, macrophages, and TINs. K, one or more antibodies that activate TIL or MIL, cytokines and / or T It further contains stimulating molecules. For example, antibodies include anti-CD28, CD3, B7-1, B7-2, and anti-inducing antibodies. Conductive costimulatory factor (ICOS), ICOS ligand, anti-CD27, CD70, 4-1BBL , anti-41-BB, anti-CD40L, CD40, anti-DAP10, anti-CD30, CD30L, anti- TIM-1, anti-TIM-2, anti-TIM-3, anti-CD44, anti-NK1.1, lectin-like transcription Substance-1 (LLT-1), anti-CD137, CD48, MICA, anti-2B4 and anti-glucoco It can contain luticoid-induced tumor necrosis factor receptor-associated protein (GITR), Tocaines are IL-2, IL-7, IL-15, IL-21, TNF-α or IFN- It can contain gamma.

[0094] D. Methods of treating cancer 68. In one embodiment, the bioresponsive hydrates disclosed herein are Including administration to any of the Rogel Matrix targets cancer or metastasis in the target. Methods for treating, preventing, inhibiting, and / or reducing migration. For example, disclosed herein. This involves administering a hydrogel matrix containing one or more chemoattractants to the subject. A method of treating cancers that include the subject, and one or more chemoattractants, CC motif chemo Caine ligand (CCL) 1 (CCL1), CCL5, CCL19, CCL21, CCL 22, CCL28, CXC motif chemokine ligand (CXCL)1 (CXCL1 ), CXCL9, CXCL10, CXCL11, or CXCL CXCLCXCL12 , M-CSF, GM-CSF, MCP-1, MCP-3, CCL2, CCL3, CCL7 , CCL20, CX3CL1, BRAK, IL-12, S1P, and / or MCP2 The chemoattractant contains 12, which attracts immune cells (e.g., T cells, NK cells, NK cells) to the hydrogel. It attracts and retains T cells, dendritic cells, macrophages, TINKs, TILs, or MILs. do.

[0095] 69. In one embodiment, the treatment, prevention, or inhibition of cancer or metastasis disclosed herein The hydrogel matrix used in the method of reducing and / or reducing the chimeric antigen receptor ( CAR), NK cell receptor (anti-CD3 antibody, CD1d, immunoglobulin Fc fragment This includes, but is not limited to, an anti-Fcγ receptor (FcγRIII) antibody, N KT cell receptor or T cell receptor (TCR) (including antigen-specific T cell receptors, but this The invention may further include a viral vector encoding (not limited to) the above. The viral vector transduces immune cells, and these transduced immune cells then develop a hydrogen It is released from the virus against cancer. The viral vector is administered to the target before or against Approximately 1 to 14 days after administering the hydrogel matrix to elephants, It is understood and intended that it can be introduced into S. Viral vectors can be introduced into hydrogels in vivo. For example, The rus vector is used during and after the gelation of the hydrogel, but only when the hydrogel is administered to the target. Before, or after administering the hydrogel to the subject, 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 On the 0th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, or 21st And it can be implemented.

[0096] 70. The disclosed chemiactive substances may leach into the hydrogel or release over time. It is released and attracts immune cells to the hydrogel, and those immune cells (e.g., T cells, NK cells, N) KT cells, dendritic cells, macrophages, TINK, TIL or MIL) are transgenes. (For example, chimeric antigen receptors, T cell receptors, NK cell receptors and / or NK T cell receptors) It is understood that transduction can occur by cellular receptors, and is intended to be used herein. In one embodiment, the release of the chemical attractant occurs from 1 hour after administration of the hydrogel. This can be approximately 12 weeks after administration. For example, the release of a chemoattractant occurs after the administration of the hydrogel. After, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 1 6, 17, 18, 19, 20, 21, 22, 23, 24, 28, 30, 36, 42, 48 , 60, 72 hours, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 1 6, 17, 18, 19, 20, 21, 28, 30, 31, 5, 6, 7, 8, 9, 10, It may be 11 or 12 weeks. In one embodiment, the cancer disclosed herein is or a method for treating, preventing, inhibiting and / or reducing metastasis, comprising one or more chemotaxis The substance is released approximately 1 hour after hydrogel administration to approximately 12 weeks after hydrogel administration. The release time of the attractant depends on whether the chemical attractant is released before or after gelation into the hydrogel polymer. It can be adjusted by adding to the following, and in this case, the addition after gelation is a chemical attractant. This provides a rapid release time for quality, and also facilitates the attachment of the polymer during or before gelation. Adding it results in a delayed release.

[0097] 71. Similarly, one advantage of the disclosed hydrogel over conventional CAR therapy is C AR T cells, CAR NK cells, CAR NK T cells, CARMA, TINK, MI It is understood that L or TIL is a slow divergence into the tumor microenvironment, and also in this specification Even if they exist, it is intended. Immune cells (e.g., T cells, NK cells, NK T cells, dendritic cells, ma Release of clophages (TINK, TIL, or MIL) occurs 1 hour after administration of the hydrogel. It is understood that this can be approximately 12 weeks after administration of the hydrogel, and also in this specification This is intended. For example, the release of chemoattractants occurs approximately 1, 2, 3, 4 times after administration of the hydrogel. , 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24, 28, 30, 36, 42, 48, 60, 72 hours, 4 , 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 28, 30, 31 days, 5, 6, 7, 8, 9, 10, 11 or 12 weeks and It is possible. Therefore, in one aspect, what is disclosed herein is cancer or metastasis. A method of treating, preventing, inhibiting and / or reducing migration, in which immune cells are administered a hydrogel. It is released approximately 1 to 12 weeks after administration.

[0098] 72. Disclosed in a manner that treats, prevents, inhibits and / or reduces cancer or metastasis. The disclosed hydrogel matrix used contains one or more immunoblocking inhibitors and / It is understood that this may further include chemotherapeutic agents, and is intended to be used herein. The chemotherapeutic agents that may be used in the disclosed hydrogel matrix include: It may be any chemotherapy agent, but is not limited to the following: Abemaciclib, Abiletorone acetate, Abitrexat e(Methotrexate), Abraxane(Paclitaxel albumin) (Stabilized nanoparticle formulations), ABVD, ABVE, ABVE-PC, AC, AC-T, Adc etris (Brentuximab Vedotin), ADE, Ado-Trast uzumab Emtansine, Adriamycin (Doxorubicin salt) (Salt), Afatinib dimaleate, Afinitor (Everolim (us), Akynzeo (Netupitant and Palonosetron hydrochloride) ), Aldara (Imiquimod), Aldesleukin, Alecensa (Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed 2 sodium), Aliqopa (Copanlisib hydrochloride) ), Alkeran for injection (Melphalan hydrochloride), Alkeran tablets (Mel phalan), Aloxi (Palonosetron hydrochloride), Alunbrig ( Brigatinib), Ambochlorin (Chlorambucil), Am boclorin (Chlorambucil), Amifostine, AminoRebri Anastrozole, Aprepitant, Aredia (Pamidro nate2 sodium), Arimidex (Anastrozole), Aromas in(Exemestane), Arranon(Nelarabine), Arsen ic Trioxide, Arzerra (Ofatumumab), Erwinia Chrysanthemi asparaginase, Atezolizumab, Avasti n(Bevacizumab), Avelumab, Axitinib, Azaciti dine, Bavencio (Avelumab), BEACOPP, Becenum ( Carmustine)、Beleodaq(Belinostat)、Belinos father、BEP、BEP、No response(Inotuzu mab Ozogamycin)、Bevacizumab、Bexarotene、B edge (TositumabおよびIodine I 131 Tositum omab)、Bicalutamide、BiCNU(Carmustine)、Wheat omycin、Blinatumomab、Blincyto(Blinatumoma b)、Bortezomib、Bosulif(Bosutinib)、Bosutin ib、Brentuximab Vedotin、Brigatinib、BuMel、 Busulfan、Busulfex(Busulfan)、Cabbage、 Cabometyx(Cabozantinib-S-マレエート)、Cabozant inib-S-マレエート、CAF、Campath(Alemtuzumab)、Ca mptosar、(Irinotecan)、Capecitabine、CAP OX、Character(Fluorouracil-Fluorouracil)、Carboplatin、C ARBOPLATIN-TAXOL、Carfilzomib、Carmubris(C armustine)、Carmustine、Carmustine Implant 、Casodex(Bicalutamide)、CEM、Ceritinib、Cer ubidine (Daunorubicin or HPV)、Cervarix (HPV) More information)、Cetuximab、CEV、Chlorambucil、CHLOR AMBUCIL-PREDNISONE、CHOP、Cisplatin、Classes bine, Clafen (Cyclophosphamide), Clofarabin e, Clofarex (Clofarabine), Clolar (Clofarabi) ne), CMF, Cobimetinib, Cometriq (Cabozantini bS-maleate), Copanlisib hydrochloride, COPDAC, COPP, COP P-ABV, Cosmegen (Dactinomycin), Cotellic (Co bimetinib), Crizotinib, CVP, Cyclophosphami de, Cyfos (Ifosfamide), Cyramza (Ramucirumab) ), Cytarabine, Cytarabine liposome, Cytosar-U(C ytarabine), Cytoxan (Cyclophosphamide), Dab rafenib, Dacarbazine, Dacogen (Decitabine), Dactinomycin, Daratumumab, Darzalex (Daratu mumab), Dasatinib, Daunorubicin hydrochloride, Daunoru bicin hydrochloride and cytarabine liposomes, decitabine, De fibrotide sodium, Defitelio (Defibrotide sodium) Degarelix, Denileukin Diftitox, Denosum ab, DepoCyt (Cytarabine liposome), Dexamethasone e, Dexrazoxane hydrochloride, Dinutuximab, Docetaxel, D oxil (Doxorubicin hydrochloride liposome), Doxorubicin hydrochloride Doxorubicin hydrochloride liposomes, Dox-SL (Doxorubicin salt Salt acid liposomes), DTIC-Dome (Dacarbazine), Durvalum ab, Efudex (Fluorouracil - topical), Elitek (Rasbu ricase), Ellence (Epirubicin hydrochloride), Elotuzuma b, Eloxatin (Oxaliplatin), Eltrombopag Olam ine, Emend(Aprepitant), Empliciti(Elotuzum) ab), Enasidenib mesylate, Enzalutamide, Epirubi cin hydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge (Vismodegib), Erlotinib hydrochloride, Erwinaze (Erwinia chrysanthemi asparaginase), E thyol (Amifostine), Etopophos (Etoposide Ph osphate), Etoposide, Etoposide Phosphate, E vacet (Doxorubicin hydrochloride liposome), Everolimus, Ev ista, (Raloxifene hydrochloride), Evomela (Melphalan hydrochloride) Salt), Exemestane, 5-FU (for Fluorouracil injection), 5-FU (Fluorouracil - topical use), Fareston (Toremifene), Farydak (Panobinostat), Faslodex (Fulvestra nt), FEC, Femara (Letrozole), Filgrastim, Flu dara (Fludarabine phosphate), Fludarabine Phos phate, Fluoroplex (Fluorouracil - topical use), Fluor ouracil Injection, Fluorouracil - Topical Use, Fluta mide, Folex (Methotrexate), Folex PFS (Metho trexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOL FIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU-LV, Fulvestrant, Gardasi l (recombinant HPV tetravalent vaccine), Gardasil 9 (recombinant HPV nonavalent vaccine) , Gazyva (Obinutuzumab), Gefitinib, Gemcitab ine hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINE -OXALIPLATIN, Gemtuzumab Ozogamicin, Gemza r(Gemcitabine hydrochloride), Gilotrif(Afatinib dimaleate) Gleevec (Imatinib mesylate), Gliadel (Carmus) Carmustine implant), Gliadel wafer (Carmustine implant) (Lactic acid), Glucarpidase, Goserelin acetate, Halaven (Eribulin mesylate), Hemangeol (Propranolol hydrochloride) ), Herceptin (Trastuzumab), HPV bivalent vaccine, recombinant, H PV nonavalent vaccine, recombinant, HPV tetravalent vaccine, recombinant, Hycamtin (Topo tecan hydrochloride), Hydrea (Hydroxyurea), Hydroxyure a, Hyper-CVAD, Ibrance (Palbociclib), Ibritu momab Tiuxetan, Ibrutinib, ICE, Iclusig(Pon atinib hydrochloride, Idamycin (Idarubicin hydrochloride), Idaru bicin hydrochloride, Idelalisib, Idhifa (Enasidenib mesile Ifex (Ifosfamide), Ifosfamide, Ifosfami dum(Ifosfamide), IL-2(Aldesleukin), Imatin ib mesylate, Imbruvica (Ibrutinib), Imfinzi (Dur Valumab), Imiquimod, Imlygic (Talimogene La herparepvec), Inlyta (Axitinib), Inotuzumab Ozogamicin, Interferon Alfa-2b, recombinant, interro Ikin-2 (Aldesleukin), Intron A (recombinant interferon α-2) b) Iodine I 131 Tositumomab and Tositumoma b, Ipilimumab, Iressa (Gefitinib), Irinoteca n hydrochloride, Irinotecan hydrochloride liposome, Istodax (Romideps) in), Ixabepilone, Ixazomib Citrate, Ixempra (Ixabepilone), Jakafi (Ruxolitinib phosphate), JEB, Jevtana (Cabazitaxel), Kadcyla (Ado-Tra stuzumab emtansine), Keoxifene (Raloxifene) Hydrochloride), Kepivance (Palifermin), Keytruda (Pemb) rolizumab), Kisqali (Ribociclib), Kymriah (T isagenlecleucel), Kyprolis (Carfilzomib), L anreotide acetate, lapatinib ditosylate, Lar truvo(Olaratumab), Lenalidomide, Lenvatini β-mesylate, Lenvima (Lenvatinib mesylate), Letrozol e, Leucovorin Calcium, Leukeran(Chlorambuc il), Leuprolide acetate, Leustatin (Cladribin e), Levulan (Aminolevulinic Acid), Linfoliz in(Chlorambucil), LipoDox(Doxorubicin hydrochloride) Posome), Lomustine, Lonsurf(Trifluridine and Tipiracil hydrochloride, Lupron (Leuprolide acetate), Lupron Depot (Leuprolide acetate), Lupron De pot-Ped (Leuprolide acetate), Lynparza (Olapa rib), Marqibo (Vincristine Sulfate liposome), M a tulane (Procarbazine hydrochloride), Mechlorethamine salt Salts, Megastrol acetate, Mekinist (Trametinib), Melphalan, Melphalan hydrochloride, Meraptopurine, Me sna, Mesnex (Mesna), Methazolastone (Temozol) omide), Methotrexate, Methotrexate LPF(Met hotrexate), Methylnaltrexone Bromide, Mexa te(Methotrexate), Mexate-AQ(Methotrexate) Midostaurin, Mitomycin C, Mitoxantrone hydrochloride, Mi tozytrex (mitomycin C), MOPP, Mozobil (Plerixaf) or), Mustargen (Mechlorethamine hydrochloride), Mutamy cin (Mitomycin C), Myleran (Busulfan), Mylosar ( Azacitidine), Mylotarg (Gemtuzumab Ozogami) cin), Nanoparticle Paclitaxel (Paclitaxel a Lubumin-stabilized nanoparticle formulation), Navelbine (Vinorelbine supplement) Acid ester), Necitumumab, Nelarabine, Neosar (Cyc lophosphamide), Neratinib Maleate, Nerlynx (Neratinib maleate), Netupitant and Palonosetr on hydrochloride, Neulasta (Pegfilgrastim), Neupogen (F ilgrastim), Nexavar (Sorafenib Tosylate), N ilandron (Nilutamide), Nilotinib, Nilutamide e, Ninlaro (Ixazomib Citrate), Niraparib Citrate Nivolumab monohydrate, Nolvadex (Tamoxifen citrate) ), Nplate (Romiplostim), Obinutuzumab, Odomz o(Sonidegib), OEPA, Ofatumumab, OFF, Olapari b, Olaratumab, Omacetaxine Mepesuccinate, O ncaspar (Pegaspargase), Ondansetron hydrochloride, Oni vyde (Irinotecan hydrochloride liposome), Ontak (Denileuki Diftitox), Opdivo (Nivolumab), OPPA, Osime rtinib, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized nanoparticle formulations, PAD, Palbociclib, Palifermi n, Palonosetron hydrochloride, Palonosetron hydrochloride and Netu pitant, Pamidronate 2 sodium, Panitumumab, Pan obinostat, Paraplat(Carboplatin), Paraplat in(Carboplatin), Pazopanib hydrochloride, PCV, PEB, PEG aspargase, Pegfilgrastim, Peginterferon Al fa-2b, PEG-intron (Peginterferon Alfa-2b), P embrolizumab, Pemetrexed Disodium, Perjeta (Pertuzumab), Pertuzumab, Platinol (Cisplat in), Platinol-AQ (Cisplatin), Plerixafor, Po malidomide, Pomalyst (Pomalidomide), Ponati nib hydrochloride, Portrazza (Necitumumab), Pralatrexa te, Prednisone, Procarbazine hydrochloride, Proleukin ( Aldesleukin), Prolia(Denosumab), Promacta( Eltrombopag Olamine, Propranolol hydrochloride, Prov enge(Sipuleucel-T), Purinethol(Mercaptopu rine), Purixan(Mercaptopurine), Radium 223 Dichloride, Raloxifene hydrochloride, Ramucirumab, Ra sburicase, R-CHOP, R-CVP, recombinant human papillomavirus (HP) V) Bivalent vaccine, recombinant human papillomavirus (HPV) nonavalent vaccine, recombinant human Papillomavirus (HPV) quadrivalent vaccine, recombinant interferon α-2b, Reg orafenib, Relistor (Methylnaltrexone bromide), R- EPOCH, Revlimid(Lenalidomide), Rheumatrex( Methotrexate), Ribociclib, R-ICE, Rituxan(R ituximab), Rituxan Hycela (Rituximab and human hyphae Alronidase), Rituximab, Rituximab and human hyaluronidase Ze, Rolapitant hydrochloride, Romidepsin, Romiplostim, Rubidomycin (Daunorubicin hydrochloride), Rubraca (Ruc aparib camsylate), Rucaparib camsylate, Ruxolitini β-phosphate, Rydapt (Midostaurin), Sclerosol intrapleural Talc Siltuximab Sipuleucel-T Soma tuline Depot(Lanreotide drugs) Sonidegib Sorafenib Sprycel(Dasatinib) STANF ORD V Preservatives (Talc), Steritalc (Talc), Stv arga(Regorafenib) , Sunitinib マエート , Sutent( Sunitinib Sylatron(Peginterferon A). lfa-2b), Sylvant (Siltuximab), Synribo (Omac). etaxine Anchorage Tabloid(Thioguanine)T AC、Tafinlar(Dabrafenib)、Tagrisso(Osimert inib)、Talc、Talimogene Laherparepvec、Tamo xifen.Tarabine PFS(Cytarabine) Tarc eva(Erlotinib Dosage)、Targretin(Bexarotene)、 Tasigna(Nilotinib)、Taxol(Paclitaxel)、Tax otere(Docetaxel)、Tecentriq、(Atezolizumab). )、Temodar(Temozolomide)、Temozolomide、Tem sirolimus, thalidomide, thalomide of) Thioguanine, Thiotepa, Tisagenlegleuce l, Tolak(Fluorouracil-drug), Topotecan, T.S oremifene, Torisel (Temsirolimus), Tositumo mab and iodine-131 Tositumomab, Totect(Dexrazox (ane hydrochloride), TPF, Trabectedin, Trametinib, Trast uzumab, Treanda (Bendamustine hydrochloride), Trifluri dine and Tipiracil hydrochloride, Trizenox (arsenic trioxide), Tyke rb (Lapatinib ditosylate), Unituxin (Dinutuximab ), Uridine Triacetate, VAC, Vandetanib, VAMP Varubi (Rolapitant hydrochloride), Vectibix (Panitum mab), VeIP, Velban(Vinblastine Sulfate), Ve lcade (Bortezomib), Velsar (Vinblastine sulfate) Vemurafenib, Venclexta (Venetoclax), Vemurafenib netoclax, Verzenio (Abemaciclib), Viadur (Le uprolide acetate, Vidaza (Azacitidine), Vinb lastine sulfate, Vincasar PFS (Vincristine sulfate Vincristine sulfate, Vincristine sulfate Tryposomes, Vinorelbine tartrate, VIP, Vismodegib , Vistogard (Uridine triacetic acid), Voraxaze (glucarpidase) ), Vorinostat, Votrient (Pazopanib hydrochloride), Vyxe os (Daunorubicin hydrochloride and Cytarabine liposomes), We llcovorin (Leucovorin calcium), Xalkori (Crizo tinib), Xeloda (Capecitabine), XELIRI, XELOX , Xgeva (Denosumab), Xofigo (Radium 223 Dich loride), Xtandi (Enzalutamide), Yervoy (Ipil) imumab), Yondelis (Trabectedin), Zaltrap (Zi v-Aflibercept), Zarxio (Filgrastim), Zejula (Niraparib tosylate monohydrate), Zelboraf (Vemurafeni b), Zevalin (Ibritumomab Tiuxetan), Zinecar d(Dexrazoxane hydrochloride), Ziv-Aflibercept, Zofran (Ondansetron hydrochloride), Zoladex (Goserelin acetate) ), Zoledronic Acid, Zolinza (Vorinostat), Zo meta(Zoledronic Acid), Zydelig(Idelalisib ), Zykadia (Ceritinib), and / or Zytiga (Avilatero) Examples include (Abiraterone acetate). Checkpoint inhibitors and However, it is not limited to PD-1 (Nivolumab (BMS-93) 6558 or MDX1106), CT-011, MK-3475), PD-L1 (MD X-1105(BMS-936559), MPDL3280A, MSB0010718C ), PD-L2(rHIgM12B7), CTLA-4(ipilimumab(Ipilimu mab)(MDX-010), Tremelimumab (CP-6) 75,206)), IDO, B7-H3(MGA271), B7-H4, TIM3, LA An example is an antibody that blocks G-3 (BMS-986016).

[0099] 73. The disclosed composition is effective in treating any disease in which unregulated cell proliferation occurs, such as cancer. It can be used to treat cancers that can be treated using the disclosed composition, for example. A descriptive and non-definitive list is as follows: lymphoma, B-cell lymphoma, T-cell lymphoma Pancreas, mycosis fungoides, Hodgkin's disease, myeloid leukemia, bladder cancer, brain tumor, nervous system cancer, head and neck Cancer, lung cancer including squamous cell carcinoma of the head and neck, small cell lung cancer and non-small cell lung cancer, and neurological cancer. Transblastoma / gliablastoma, ovarian cancer, skin cancer, liver cancer, melanoma, mouth, throat, larynx, and Squamous cell carcinoma of the lung, cervical cancer, breast cancer, and epithelial cancer, kidney cancer, urinary tract cancer Genitourinary cancer, lung cancer, esophageal cancer, head and neck cancer, colorectal cancer, hematopoietic cancer; testicular cancer; colon Cancer, colorectal cancer, prostate cancer, or pancreatic cancer. [Examples]

[0100] E. Examples 74. The following examples illustrate the compounds, compositions, articles, and devices claimed herein. A full disclosure and description of how the and / or methods are made and evaluated is to be made to those skilled in the art. This information is presented for illustrative purposes only and is intended to be purely illustrative. It is not intended to be precise. Accuracy regarding numbers (e.g., quantity, temperature, etc.) is not guaranteed. Efforts have been made to achieve this, but some errors and deviations should be taken into consideration. Unless otherwise specified, parts refer to parts by weight, temperature is in degrees Celsius or ambient temperature, and pressure is large. Atmospheric pressure or close to atmospheric pressure.

[0101] 1. Example 1: Production of a microporous gel via cryogel formation 75. Crosslink the alginate with calcium gluconate and freeze it at -20°C. Gelation occurs near the ice crystals, and these ice crystals act as pologens (Figure 2). Chloroporosis promotes chemokine-mediated T cell infiltration, and also by viral particles. It provides an interface for transduction. Figure 3 shows the gel prepared in this way. The image shows a scanning electron microscope photograph.

[0102] 2. Example 2: Host T cell-mediated recruitment and reprogramming 76. In this embodiment, the transplanted CXCL10-releasing macroporous scaffold was transplanted It is demonstrated that T cells can be efficiently recruited (Figures 4A and 4B). In this embodiment, Approximately 20% of activated human T cells preloaded onto the alginate scaffold become viral vectors - Transduction occurs by (Figures 5A and 5B), and CD3 / CD3 occurs within the gel. When 28 antibodies and IL-2 are combined, 2-3 million human PB cells are transplanted. 1-2% of MCs are recruited from the bloodstream (Figures 5C and 5D), and CD3 / CD28 antibodies and It has been demonstrated that IL-2 provides environmental factors that promote T cell activation.

[0103] 3. Example 3: Optimal parameters in T cell reprogramming mediated by scaffolds Optimize 77. The results showed that controlled release of the chemoattractant CXCL10 leads to the release of circulating T cells. It is demonstrated that they are mobilized to the plant scaffold. Furthermore, activated T cells pre-lot This specification shows that the modified scaffold can be transduced with a virus. GFP and In vivo phenotype of viral vectors against recruited T cells using CAR-T vectors We will consider the efficiency of implementation.

[0104] 4. Example 4: In vivo transduction of T cells using a viral vector encoding GFP Optimization and Feature Analysis 78. Test the ability of encapsulated retroviral vectors to transduce mobilized T cells. It is possible to do this. Human PBMC (1 × 10) in NSG mice. 7 Cells / mouse) transferred for 20 days It can be implanted. It encodes CXCL10 (4 μg / mg arginate) and GFP. An alginate scaffold loaded with retroviral vectors (two different concentrations) is used. It can be transplanted into the subcutaneous space of the mortar (n=4). At several points in time (after the scaffolding is transplanted). On days 4, 6, and 8, the scaffold can be explanted, and the frequency of CD45+CD3+GFP+ cells increases. The degree can be evaluated by flow cytometry. The following are included as controls: Possible methods include: 1) an empty gel without chemokines, and 2) administration of a pseudovirus.

[0105] 5. Example 5: In-situ generation of CAR-T cells: 79. Use chemokine-loaded scaffolding and viral vectors encoding CAR. This allows CAR-transduction to be performed in situ, enabling optimal recruitment of T cells. And the reprogramming conditions can be tested with a virus construct that codes for CAR. n=10 is used (see the "Statistical Analysis" section). The scaffolding is to be explanted. This can be done, and the percentage of recruited cells and CD19.CAR+ cells is flusa It can be evaluated by iteometry. The following can be included as controls: 1) Empty gel without chemokines, and 2) administration of a pseudovirus. The viral vector is In Nvivo, it can be broken down over time. In such a scenario, the virus can be degraded once It can be administered via the slow infusion within the scaffold described above. Similar results have been observed with GFP and C It can also be observed in viral vectors that encode AR.

[0106] 80. For Hu-PBMC-NSG mice, a scaffold (CCI-A) containing CXCL10 was used. lg) was inserted. The gamma retrovirus encoding human CD19.CAR was placed on a slow scaffold. The virus was administered by internal injection. Three days after virus administration, the scaffold was explanted, digested, and isolated. The cells were stained with CD3 and CD19 antibodies and analyzed by flow cytometry. (Figure 6). Figure 7 shows the in vivo phenotypic induction of T cells mobilized by CD19 CAR virus. It indicates entry.

[0107] 6. Example 6: Characteristic analysis of the migration of transduced T cells: 81. After T cell recruitment and reprogramming, the reprogrammed cells enter the bloodstream. The effective release of transduced T cells into the circulating system is crucial for therapeutic function. The mobility efficiency and dynamics can be analyzed using feature analysis.

[0108] 82. To evaluate the migration of transduced T cells into the bloodstream, T cell recruitment and morphology GFP+ or CAR+T cells in mice (n=12) carrying scaffolds for quality introduction The cells can be measured at several points in time (5, 10, and 15 days after scaffold implantation). By killing mice, it is possible to quantify the expression of GFP or CAR in systemic T cells. It is possible. The best conditions can be used for T cell recruitment and reprogramming. At several points, the blood, spleen, bone marrow, draining area, and peripheral lymph nodes are examined. T cells were isolated and analyzed by flow cytometry to determine CD45+CD3+GFP+ and This can assess the frequency and number of CD45+CD3+CAR+ cells. As a control... This can include: 1) a virus-free scaffold, 2) GFP+ T cells pre-treated. The seeded scaffold. The macroporous property of the scaffold allows for the transfer of CAR-T cells from the scaffold to the bloodstream. Promotes movement. If movement is slow, or if T cells are trapped within the scaffold, phosphorus As it is known that pocytes move along collagen fibers, the scaffold is collagen-like It can be modified with butyl. Furthermore, alginates can be carefully chemically modified to produce The decomposition rate can be adjusted, and the release of CAR-T can be increased by utilizing the decomposition of the scaffolding. It can also be done.

[0109] 7. Example 7: Statistical Analysis: 83. The number of animals for the GFP experiment was determined based on 80% power, with a two-sided α = 0.05. The above difference was detected. The number of animals required for in situ generation of CAR T cells is CD19 The determination is based on a two-sided 95% confidence interval (CI) for the mean percentage, and in doing so, The full width at half maximum of the CI was approximately 3.6%, and the estimated standard deviation was approximately 5%.

[0110] 8. Example 8: Tumor Model 84. Daudi.ffluc cells (CD19+ tumor cells) are used, N Using SG mice, a mouse model of human Burkitt lymphoma (a type of B-cell lymphoma) was developed. We developed a model (Figure 8). Mice were given tumors on 19, 24, 29, 33, and 43 days after tumor inoculation. As shown in Figure 9A, the tumor size of mice was observed by luminescence, and the untreated mice were... Comparison between control mice and mice treated with an alginate scaffold or those receiving IV. We also measured the weight gain and flux of the mice (Figure 9B), but the treatment group was different. No discernible difference was shown. Furthermore, IV administration of CD19 CAR T cells In animals that have been given or treated with CCI-alginate scaffolding (CCI-Alg), tumor contact At several post-seeding time points, CCI scaffolds were transplanted, or CAR-T cells were implanted. v. Number of CAR-T cells in 100 μl of injected mouse blood. Blood was collected from the cheek of the mouse. The red blood cells are hemolyzed, and the cells are stained with Hu-CD45, Hu-CD3, and CAR.19 antibodies. The color was determined and analyzed by flow cytometry (Figure 10).

[0111] F. References Brentjens,RJet al.CD19-targeted T cel ls rapidly induce molecular remissions i n adults with chemotherapy-refractory ac ute lymphoblastic leukemia.Sci.Transl.Me d.5,177ra38(2013). Dotti, G., Gottschalk, S., Savoldo, B. & Brenn er,MKDesign and development of therapy es using chimeric antigen receptor-expre ssing T cells.Immunol.Rev.257,107-126(20 14). Enblad,G.et al.A PHASE I / IIa TRIAL USIN G CD19-TARGETED THIRD GENERATION CAR T C ELLS FOR LYMPHOMA AND LEUKEMIA.Clin.Canc er Res.(2018).doi:10.1158 / 1078-0432.CCR- 18-0426 Grupp,S.A.et al.Chimeric Antigen Recept or-Modified T Cells for Acute Lymphoid L eukemia.N.Engl.J.Med.368,1509-1518(2013) . Kalos,M.et al.T cells with chimeric ant igen receptors have potent antitumor eff ects and can establish memory in patient s with advanced leukemia.Sci.Transl.Med. 3,95ra73(2011). Kochenderfer,J.N.&Rosenberg,S.A.Treatin g B-cell cancer with T cells expressing anti-CD19 chimeric antigen receptors.Nat .Rev.Clin.Oncol.10,267-276(2013). Kochenderfer,J.N.et al.Eradication of B -lineage cells and regression of lymphom a in a patient treated with autologous T cells genetically engineered to recogni ze CD19.Blood 116,4099-4102(2010). Lee,D.W.et al.T cells expressing CD19 c himeric antigen receptors for acute lymp hoblastic leukaemia in children and youn g adults:a phase 1 dose-escalation trial .Lancet 385,517-528(2015). Maude,S.L.et al.Chimeric antigen recept or T cells for sustained remissions in l eukemia.N.Engl.J.Med.371,1507-1517(2014) . Prasad,V.Immunotherapy:Tisagenlecleucel -the first approved CAR-T-cell therapy: implications for payers and policy maker s.Nat.Rev.Clin.Oncol.15,11-12(2018). Ramos,C.A.,Savoldo,B.&Dotti,G.CD19-CAR trials.Cancer J.20,112-118(2014). Sadelain,M.,Brentjens,R.&Riviere,I.The promise and potential pitfalls of chimer ic antigen receptors.Curr.Opin.Immunol.2 1,215-223(2009). Sharma,P.,King,G.T.,Shinde,S.S.,Purev,E .&Jimeno,A.Axicabtagene ciloleucel for t he treatment of relapsed / refractory B-ce ll non-Hodgkin’s lymphomas.Drugs Today 5 4,187-198(2018).

Claims

1. It contains one or more chemoattractants, and the one or more chemoattractants are C-C motif chemoattractants. Cain ligand (CCL) 1 (CCL1), CCL5, CCL19, CCL21, CCL 22. CCL28, C-X-C motif chemokine ligand (CXCL) 1 (CXCL1 ), CXCL9, CXCL10, CXCL11, CXCL12, M-CSF, GM-CS F, MCP-1, MCP-3, CCL2, CCL3, CCL7, CCL20, CX3CL 1. A hydrogerm containing BRAK, IL-12, S1P, and / or MCP2. .

2. Chimeric antigen receptor (CAR), NK cell receptor, NK T cell receptor, or T cell receptor The hydrogermatric according to claim 1, further comprising a viral vector encoding a body. vinegar.

3. The aforementioned viral vector is a lentivirus, retrovirus, adenovirus or The hydrogel matrix according to claim 2, comprising deno-associated virus.

4. T cells, macrophages, natural killer (NK) cells, NK T cells, tumor infiltration N Activating K cells (TINK), tumor-infiltrating lymphocytes (TIL), or medullary lymphocytes (MIL) A claim further comprising one or more antibodies, cytokines, and / or costimulatory molecules that are conditioned. A hydrogel matrix as described in any one of items 1 to 3.

5. The antibodies include anti-CD3, CD28, B7-1, B7-2, anti-inducible co-stimulatory factor (ICO S), ICOS ligand, anti-CD27, CD70, 4-1BBL, anti-41-BB, anti-CD 40L, CD40, anti-DAP10, anti-CD30, CD30L, anti-TIM-1, anti-TIM- 2. Anti-TIM-3, anti-CD44, anti-NK1.1, lectin-like transcription factor-1 (LLT-1) , anti-CD137, CD48, MICA, anti-2B4, and anti-glucocorticoid-induced tumor The hydrogermatrix according to claim 4, comprising necrosis factor receptor-associated protein (GITR). .

6. The cytokines mentioned above include IL-2, IL-7, IL-15, IL-21, TNF-α, and The hydrogel matrix according to claim 4, wherein the matrix contains IFN-γ.

7. A hydrogermatic according to any one of claims 1 to 6, further comprising a chemotherapeutic agent. vinegar.

8. To administer the hydrogel matrix according to any one of claims 1 to 7 A method for treating cancer in the subject mentioned above.

9. The procedure involves administering a hydrogel matrix containing one or more chemoattractants to the subject. A method for treating cancer in the subject, wherein one or more chemoattractants are C-C motif Dandruff-mokein ligand (CCL) 1 (CCL1), CCL5, CCL19, CCL21, CCL22, CCL28, C-X-C motif chemokine ligand (CXCL) 1 (CX CL1), CXCL9, CXCL10, CXCL11, CXCL12, M-CSF, GM - CSF, MCP-1, MCP-3, CCL2, CCL3, CCL7, CCL20, CX The chemical agent comprises 3CL1, BRAK, IL-12, S1P, and / or MCP2. A method for treating cancer, wherein an attractant attracts and retains immune cells in the hydrogel.

10. The immune cells include T cells, NK cells, NK T cells, macrophages, dendritic cells, TI A method for treating cancer according to claim 9, comprising NK, TIL, or MIL.

11. Claim 9 or 1, wherein the hydrogel matrix further comprises an immunoblocking inhibitor. Cancer treatment methods as described in section 0.

12. Any of claims 9 to 11, wherein the hydrogel matrix further comprises a chemotherapeutic agent. The cancer treatment methods described in item 1.

13. The aforementioned hydrogel is a chimeric antigen receptor (CAR), NK cell receptor, and NK T cell receptor. Claims 9 to 12 further include a viral vector encoding a body or T cell receptor. A cancer treatment method described in either item.

14. The viral vector transduces the immune cells, and the transduced immune cells The method for treating cancer according to claim 13, wherein the cancer is released from the hydrogel to the cancer. 。

15. The viral vector is administered to the subject approximately 1 to 14 days after administration of the hydrogel. The method for treating cancer according to claim 14, wherein the hydrogel is introduced in vivo.

16. The viral vector is introduced into the hydrogel before the hydrogel is administered to the target. The method for treating cancer according to claim 14.

17. Claim 1, wherein the immune cells are released for approximately 1 to 12 weeks after administration of the hydrogel. A cancer treatment method described in any one of items 4 to 16.

18. The one or more chemical attractants present approximately 1 hour after administration of the hydrogel. A method for treating cancer according to any one of claims 9 to 17, wherein the cancer is released approximately 12 weeks later.

19. A method for transducing target immune cells, wherein the target is subjected to one or more chemoattractants and , comprising administering a hydrogel containing a viral vector encoding a transgene, A method for introducing immune cells into the body.

20. The viral vector is administered to the subject approximately 1 to 14 days after administration of the hydrogel. The immunocellular cells described in claim 19 are transduced in vivo into the hydrogel. method.

21. The viral vector is introduced into the hydrogel before the hydrogel is administered to the target. A method for transducing immune cells according to claim 19, wherein the immune cells described in claim 19 are introduced.

22. The one or more chemical attractants are C-C motif chemokine ligands (CCL) 1(C CL1), CCL5, CCL19, CCL21, CCL22, CCL28, C-X-C model Chief chemokine ligand (CXCL) 1 (CXCL1), CXCL9, CXCL10, CXCL11, CXCL12, M-CSF, GM-CSF, MCP-1, MCP-3, C CL2, CCL3, CCL7, CCL20, CX3CL1, BRAK, IL-12, S1 The immune cells according to any one of claims 19 to 21, comprising P and / or MCP2. A method for introducing traits.

23. The one or more chemical attractants present approximately 1 hour after administration of the hydrogel. Transducing immune cells according to any one of claims 19 to 22, which are released approximately 12 weeks later. How to do it.

24. The transgene encoded by the viral vector is CAR, NK cell receptor A body comprising an NK cell receptor or a T cell receptor, according to any one of claims 19 to 23. A method for transducing the described immune cells.

25. The aforementioned viral vector is a lentivirus, retrovirus, adenovirus or Transduced immune cells according to any one of claims 19 to 24, including deno-associated virus. How to do it.

26. The immune cells include T cells, NK cells, NK T cells, macrophages, dendritic cells, TI Immune cells according to any one of claims 19 to 25, comprising NK, TIL, or MIL Methods for introducing traits.

27. The aforementioned hydrogel contains T cells, NK cells, NK T cells, macrophages, dendritic cells, and T One or more antibodies, cytokines, and / or that activate INK, TIL, or MIL. Transducing an immune cell according to any one of claims 19 to 26, further comprising a co-stimulatory molecule. How to do it.

28. The antibody may be anti-CD28, CD3, B7-1, B7-2, anti-inducible costimulatory factor (ICO S), ICOS ligand, anti-CD27, CD70, 4-1BBL, anti-41-BB, anti-CD 40L, CD40, anti-DAP10, anti-CD30, CD30L, anti-TIM-1, anti-TIM- 2. Anti-TIM-3, anti-CD44, anti-NK1.1, lectin-like transcription factor-1 (LLT-1) , anti-CD137, CD48, MICA, anti-2B4, and anti-glucocorticoid-induced tumor The immune cells according to claim 27, comprising necrosis factor receptor-related protein (GITR), How to install it.

29. The cytokines mentioned above include IL-2, IL-7, IL-15, IL-21, TNF-α, and A method for transducing immune cells according to claim 27, wherein the transduction is performed by IFN-γ.

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