CN111789829A

CN111789829A - Application of U-5021 compounds in preventing and treating cytokine release syndrome caused by CAR-T therapy

Info

Publication number: CN111789829A
Application number: CN201910277535.6A
Authority: CN
Inventors: 刘茂玄; 许晨光; 张宏玲
Original assignee: Shenzhen Bindebio Technology Co ltd
Current assignee: Shenzhen Bindebio Technology Co ltd
Priority date: 2019-04-08
Filing date: 2019-04-08
Publication date: 2020-10-20

Abstract

The invention provides application of a U-5021 compound in preparation of a preparation for preventing and/or treating cytokine release syndrome caused by CAR-T therapy. Also provides a pharmaceutical composition for treating malignant tumor, autoimmune disease or AIDS, which comprises chimeric antigen receptor T cells and U-5021 compounds. According to the invention, the chimeric antigen receptor T cell and the U-5021 compound are used together, so that CRS side effects caused by CAR-T cell therapy can be prevented and/or treated at a low economic cost, and a good treatment effect of the CAR-T cell therapy is further exerted.

Description

Application of U-5021 compounds in preventing and treating cytokine release syndrome caused by CAR-T therapy

Technical Field

The invention relates to the technical field of medicines, in particular to application of U-5021 compounds in preventing and treating cytokine release syndrome caused by CAR-T therapy.

Background

Malignant diseases such as cancer are diseases which seriously harm human health in the global scope and seriously affect life. Currently, the treatment methods for cancer and the like include nonspecific immunotherapy, active immunotherapy, adoptive cellular immunotherapy, and the like. Among them, adoptive cellular immunotherapy is considered as a method in which it is possible to completely eliminate cancer cells in the existing technologies. The chimeric antigen receptor T cell (CAR-T) technology, one of the latest immune cell technologies in current adoptive cell-based therapeutic technologies, is to transfuse genetically modified T lymphocytes (which can express specific chimeric antigen receptors) back into the human body, and activate the T lymphocytes in a manner independent of major histocompatibility complexes, thereby specifically killing malignant target cells such as cancer cells. The CAR-T technology has strong targeting, killing activity and persistence on malignant cells, thereby showing great application prospect in the aspect of treating malignant diseases such as cancer and the like.

Although CAR-T cell therapy has achieved good results in some clinical trials, it has also been associated with serious side effects, such as the most common Cytokine Release Syndrome (CRS) at present, resulting in significant increases in the levels of certain cytokines in the patient's blood, fever, tachycardia, hypotension, etc., and, in severe cases, multiple organ failure and death. Therefore, there is a need to inhibit cytokine release during CAR-T cell immunotherapy and alleviate/eliminate CRS side effects to improve the therapeutic efficacy of CAR-T cells.

Disclosure of Invention

In view of the above, the present invention provides a technical solution for preventing or treating CRS caused by CAR-T cell immunotherapy.

In a first aspect, the invention provides an application of the U-5021 compound in preparation of an agent for preventing and/or treating Cytokine Release Syndrome (CRS) caused by chimeric antigen receptor T (CAR-T) cell therapy.

Wherein, the U-5021 compound comprises U-5021, tautomer thereof, pharmaceutically acceptable salt or prodrug thereof and U-5021 derivative.

Wherein the structural formula of the U-5021 is shown as the following formula (I):

wherein the CAR-T cell therapy comprises use in the treatment of a malignancy, an autoimmune disease or AIDS.

Further, the malignant tumor includes one or more of leukemia, lymphoma, multiple myeloma, brain glioma, liver cancer, lung cancer, stomach cancer, esophageal cancer, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, cervical cancer and prostate cancer, but is not limited thereto.

Further, the autoimmune disease includes one or more of systemic lupus erythematosus, rheumatoid arthritis, chronic active hepatitis, scleroderma, pemphigus, dermatomyositis, ulcerative colitis, and hashimoto's thyroiditis, but is not limited thereto.

The U-5021 compound is a micromolecular drug, the price of the micromolecular drug is much lower than that of an antibody drug for inhibiting cell factors after CRS side effect occurs, and the purpose of relieving CRS side effect caused by CAR-T cell therapy can be achieved with lower economic cost; meanwhile, U-5021 can inhibit the activity of various cytokines by inhibiting the release of catecholamine substances, compared with the antibody drugs which can only inhibit the activity of one cytokine, so that the CRS can be prevented and/or treated better.

In a second aspect, the invention also provides a pharmaceutical composition comprising a chimeric antigen receptor T (CAR-T) cell and a compound of class U-5021.

Optionally, the CAR-T cells comprise CAR-T cells that target one or more of CD19, CD20, CD22, CD33, CD38, CD317, BCMA, EGFR, Mesothelin, DR5, c-met, OX40, HER 2.

Wherein the T cells in the CAR-T cells are from autologous T cells, allogeneic T cells, or iPSC-induced T cells.

In the pharmaceutical composition, the CAR-T cells and the U-5021 compound can be independently present or present in a mixture. When used as a pharmaceutical combination, the two may be administered simultaneously, sequentially or in different modes of administration.

In one embodiment of the invention, the CAR-T cells are administered intravenously and the compound of class U-5021 is administered intraperitoneally.

In a third aspect, the invention also provides an application of the pharmaceutical composition in preparing a preparation for treating malignant tumor, autoimmune disease or AIDS.

According to the invention, the CAR-T cell and the U-5021 compound are combined, so that CRS side effects (not only inhibiting the level of a cytokine) caused by CAR-T cell therapy can be prevented and/or treated, and a better treatment effect of CAR-T cell therapy is further exerted.

Detailed Description

While the following is a description of the preferred embodiments of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.

The embodiment of the invention provides application of a U-5021 compound in preparing a preparation for preventing and/or treating Cytokine Release Syndrome (CRS) caused by chimeric antigen receptor T (CAR-T) cell therapy.

the "U-5021 compound" of the present invention may include U-502, its tautomer, or its pharmaceutically acceptable salt or prodrug, and various derivatives that can be modified by simple chemical methods (such as reduction, substitution, etc.) on the side chain of the backbone by those skilled in the art, and still remain for preventing and/or treating CRS caused by CAR-T cell therapy, and the synthesis method of these derivatives can be referred to the textbook in the prior art.

Wherein, "pharmaceutically acceptable salt" refers to a salt of a compound prepared from a pharmaceutically acceptable non-toxic acid (including inorganic acids, organic acids), non-toxic base (including inorganic bases, organic bases), solvate, hydrate or clathrate.

Prodrugs can be prepared in situ during isolation and purification of the compound, or by separately reacting the purified compound with a suitable derivatizing agent. For example, the hydroxy group of the tautomer can be converted to an ester group by treatment with a carboxylic acid in the presence of a catalyst. Examples of cleavable alcohol prodrug moieties include: substituted or unsubstituted, branched or unbranched lower alkyl esters, such as ethyl esters, lower alkenyl esters, di-lower alkylamino lower alkyl esters, such as dimethylaminoethyl ester, acylamino lower alkyl esters, acyloxy lower alkyl esters (such as pivaloyloxymethyl ester), aryl esters, such as phenyl esters, aryl-lower alkyl esters, such as benzyl esters which may be substituted, such as aryl and aryl-lower alkyl esters substituted by methyl, halogen or methoxy, amides, lower alkylamides, di-lower alkylamides, and hydroxyamides.

The research of the applicant of the invention finds that the compound of the U-5021 can inhibit the release of the fine factors, so as to prevent and/or treat CRS caused by CAR-T cell therapy.

CAR-T cells of the invention include CAR-T cells that target a variety of different malignant disease targets, such as: targeting one or more of CD19, CD20, CD22, CD33, CD38, CD317, BCMA, EGFR, Mesothelin, DR5, c-met, OX40, HER2, and the like.

Embodiments of the present invention also provide a pharmaceutical composition comprising a chimeric antigen receptor T (CAR-T) cell and a compound of U-5021 class. The pharmaceutical composition is used for treating malignant tumor, autoimmune disease or AIDS, and can also be used for preventing and/or treating Cytokine Release Syndrome (CRS) caused by chimeric antigen receptor T (CAR-T) cell therapy.

Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is preferably chemically inert to the compounds of the present invention of class U-5021 and does not have deleterious side effects or toxicity under the conditions of use.

Such pharmaceutically acceptable carriers, e.g., excipients, stabilizers, suspending agents or diluents, and the like, are well known to those skilled in the art and are publicly available. Exemplary carriers can include water, physiological saline, aqueous dextrose and related sugar solutions, monohydric alcohols (such as ethanol, isopropanol or cetyl alcohol), glycols (such as propylene glycol or polyethylene glycol), glycerol, ketals (such as 2, 2-dimethyl-1, 3-dioxolane-4-methanol), ethers (such as poly (ethylene glycol) 400), oils, fatty acids, fatty acid esters or glycerides, acetylated fatty acid glycerides with or without pharmaceutically acceptable surfactants, such as soaps or detergents, suspending agents (such as pectin, carbomer, methyl cellulose, hydroxypropyl methyl cellulose or carboxymethyl cellulose), or emulsifying agents and other pharmaceutical adjuvants. The choice of the carrier will depend, in part, on the particular U-5021 class of compound selected, as well as the particular method used to administer the composition.

For example, the CAR-T cells are administered parenterally, e.g., intravenously, intraarterially, subcutaneously, intradermally, intrathecally, or intramuscularly. The CAR-T cells can be dissolved or suspended in a solution suitable for parenteral administration in an acceptable carrier, including isotonic sterile injectable aqueous and non-aqueous solutions. The administration modes of the U-5021 compounds comprise oral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal administration, sublingual administration, spray nasal administration and the like. When the U-5021 compound is administered by intravenous injection, subcutaneous injection, intramuscular injection, etc., it can be prepared into an injection preparation.

Where the class of U-5021 compounds is used in conjunction with CAR-T cells, various concentrations of the class of U-5021 compounds can be used in conjunction with various concentrations (or numbers) of CAR-T cells. For example, the CAR-T cells used have a density of 0.5X 10⁶～5×10⁶Individual cells/mL. For example, 1X 10⁶、2×10⁶Or 0.5X 10⁶Individual cells/mL.

In addition, the embodiment of the present invention also provides a method for treating malignant tumor, autoimmune disease or AIDS, which comprises administering the pharmaceutical composition of the present invention to a test subject with malignant target cells.

The concentration of the compound of the U-5021 class, the concentration (or number) of CAR-T cells can be adjusted according to the particular subject. Wherein the malignant target cell is a cell targeted by the CAR-T cell. Such as cells corresponding to the above-mentioned malignant tumor, autoimmune disease or AIDS.

Optionally, the CAR-T cell to malignant target cell number ratio is (2-10): 1. for example 2.5:1, 5:1 or 10: 1.

The treatment of CRS resulting from CAR-T cell immunotherapy in tumor therapy with compounds of the U-5021 class is illustrated below as a specific example. The reagents used in the present invention are all common commercial products, and are all commercially available.

The method comprises the following specific steps:

1. mixing newborn (0-2 days) SGM3(NSG Tg)^{CMV-IL3，CSF2，KITLG}1Eav/MloySzJ) mice were irradiated with sublethal doses (150cGy) and immediately injected intrahepatically with 1X 10⁵Individual human cord blood CD34+ cells, a nhsgm 3 humanized mouse was prepared. Humanized mouse T cells of nhsgm 3 were used to generate CAR-T cells targeted to CD 19.

2. Adult (6-8 weeks) SGM3(NSG Tg)^{CMV-IL3，CSF2，KITLG}1Eav/MloySzJ) mice were irradiated with a sublethal dose (200cGy) and injected immediately via tail vein with 1X 10⁵Individual human cord blood CD34+ cells, HuSGM3 humanized adult mice were prepared.

After 3.4 weeks, the adult mice in step 2 were injected 2X 10 by tail vein injection⁶CD 19-targeted CAR-T cells prepared from nHuSGM3 humanized mice in step 1.

4. Immediately after CAR-T cell injection, U-502110 mg/kg or an equal volume of PBS was i.p. daily for 3 consecutive days.

5. Weighing the body weight of the mice each day after the injection of the CAR-T cells, and measuring the body temperature of the mice; blood was taken every two days and the serum levels of human IL-6, IL-10, TNF- α and mouse serum amyloid A were measured. The CRS-related indexes are continuously detected for 21 days.

As a result, compared with the CAR-T cell therapy only targeting CD19 on the adult mouse with tumor, the CAR-T cell therapy targeting CD19 and the hydrochloride salt of U-5021 on the adult mouse with tumor have lower body temperature and lower contents of cytokines such as human IL-6, IL-10 and TNF-alpha in serum. This indicates that the use of the compounds of the U-5021 class has a significant therapeutic effect on CRS caused by CAR-T cell immunotherapy in tumor treatment.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

The application of the U-5021 compound in preparing a preparation for preventing and/or treating cytokine release syndrome caused by chimeric antigen receptor T cell therapy.
2. The use of claim 1, wherein the U-5021-like compound comprises U-5021, a tautomer thereof, a pharmaceutically acceptable salt or prodrug thereof, and a U-5021 derivative; wherein the structural formula of U-5021 is shown as the following formula (I):
3. the use of claim 1, wherein said chimeric antigen receptor T cell therapy comprises treatment of a malignancy, an autoimmune disease, or aids.
4. The use of claim 3, wherein the malignancy comprises one or more of leukemia, lymphoma, multiple myeloma, brain glioma, liver cancer, lung cancer, stomach cancer, esophageal cancer, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, cervical cancer and prostate cancer;

the autoimmune disease comprises one or more of systemic lupus erythematosus, rheumatoid arthritis, chronic active hepatitis, scleroderma, pemphigus, dermatomyositis, ulcerative colitis, and hashimoto's thyroiditis.
5. A pharmaceutical composition comprising a chimeric antigen receptor T cell and a U-5021 compound.
6. The pharmaceutical composition of claim 5, wherein the chimeric antigen receptor T cell is a chimeric antigen receptor T cell that targets one or more of CD19, CD20, CD22, CD33, CD38, CD317, BCMA, EGFR, Mesothelin, DR5, c-met, OX40, HER 2.
7. The pharmaceutical composition of claim 5, wherein the T cells used in the chimeric antigen receptor T cells are derived from autologous T cells, allogeneic T cells, or iPSC-induced T cells.
8. The pharmaceutical composition of any one of claims 5-7, wherein the chimeric antigen receptor T cells and the compound of class U-5021 are present alone or in a mixed state; the administration mode of the chimeric antigen receptor T cells and the U-5021 compound comprises simultaneous administration, sequential administration or administration in different administration modes.
9. The pharmaceutical composition of claim 8, wherein the chimeric antigen receptor T cells are administered intravenously and the class U-5021 compound is administered intraperitoneally.
10. Use of a pharmaceutical composition according to any one of claims 5-9 for the preparation of a formulation for the treatment of malignant tumors, autoimmune diseases or aids.