KR20200059161A - Pharmaceutical composition for enhancing sensitivity to cancer immunotherapy comprising IRE1alpha or XBP1 activation inhibitor - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for enhancing the sensitivity to anti-cancer immunotherapy, containing an activation inhibitor of IRE1α or XBP1 as an active component. The pharmaceutical composition for enhancing the sensitivity to anti-cancer immunotherapy of the present invention can effectively induce cancer cell death.

Description

Pharmaceutical composition for enhancing sensitivity to cancer immunotherapy comprising IRE1alpha or XBP1 activation inhibitor}

The present invention relates to a pharmaceutical composition for enhancing the sensitivity to anti-cancer immunotherapy, comprising an active inhibitor of IRE1α or XBP1 as an active ingredient.

Currently, methods of treating cancer can be largely divided into surgery, radiation therapy, chemotherapy, and biological therapy. Among these, patients who do not have easy surgery or radiation therapy (approximately 50% of all cancer cases) and those who have already metastasized cancer are mainly treated with chemotherapy.

Anti-cancer agents can be classified into first-generation chemotherapeutic agents, second-generation targeted anti-cancer agents, and third-generation immuno-cancer agents. First-generation chemotherapy drugs have serious side effects because they damage not only cancer cells but also normal cells. That is, to kill cancer cells, they attack the normal cells, destroy the patient's immune system, and due to their strong toxicity, various side effects such as hair loss, vomiting, loss of appetite, fatigue, and severe stamina occur.

Second-generation targeted anticancer drugs have the advantage of identifying only cancer cells and attacking them, but they can be used only for patients with genetic mutations, making it impossible to treat various cancers and tending to develop resistance. The 3rd generation immuno-cancer agent has a new mechanism of activating the immune cells of the human body that have been suppressed and killing cancer cells. It can be used widely in most cancers without specific genetic variation. Immune anti-cancer drugs have the effect of reducing the side effects, improving the quality of life of cancer patients, and significantly extending the survival period in that they are treated by strengthening their own immune system.

Immune anti-cancer agents exhibit anti-cancer effects by enhancing the specificity, memory, and adaptability of the immune system. In other words, since only the cancer cells are attacked accurately using the body's immune system, there are few side effects, and the immune system's memory and adaptive power are used, so patients who are effective in immunocancer drugs can see the persistent anti-cancer effect. Therefore, immuno-cancer drugs have improved side effects of first-generation chemotherapeutic drugs and resistance of second-generation targeted anti-cancer drugs, and have long-term effect, long-term survival, and broad anti-tumor activity. And low toxicity profile.

On the other hand, tumors are not only cancer cells, but also stromal cells, fibroblasts, immune and inflammatory mediator cells. It is composed of various peripheral cells such as vascular endothelial cells and connective tissue-forming cells, and they are exposed to hypoxic and acidic conditions. This special tumor microenvironment affects drug resistance to cancer treatment as well as cancer treatment by regulating the proliferation, invasion and metastasis of cancer cells. Tumor cells generally rely on aerobic glycolysis to maintain cell growth and proliferation, even under hypoxic conditions. Cancer cells are relatively resistant to growth inhibition in oxygen-deficient conditions, which is a high inducer of rapid proliferation of cancer cells. It is believed to be one of the ways to rapidly produce the cell energy and metabolites needed for anabolic activity. In order for this metabolic mechanism to be efficient, glucose must be continuously supplied to cancer cells, and due to such high dependence on glucose, cancer cells may be susceptible and sensitive to stress due to glucose deficiency. However, in cancer patients with nutrient deficiency, a lower survival rate is observed, and thus, when an anticancer agent is administered, deficiency of nutrients may make cancer cells more aggressive through cellular reactions activated by deficiency conditions rather than promoting cancer cell death. The possibility was presented. In addition, anticancer immunotherapy exhibits anticancer effects by inducing apoptosis, and nutritional stress such as glucose deficiency in tumor microenvironment of all solid cancers induces apoptosis resistance of cancer cells, so it may show resistance to anticancer immunotherapy.

In addition, according to existing studies, the concentration of glucose in solid cancer is 3 to 10 times lower than that of normal tissue. Cancer cells exposed to these tumor microenvironments for a long time may have resistance to anticancer immunotherapy even before chemoimmune therapy is performed. Therefore, there is a need to develop a drug for improving resistance to anticancer immunotherapy caused by a biochemical environmental change in which glucose is supplied in a small amount among changes in nutritional status occurring in solid cancer.

Against this background, the present inventors, when combined with an anti-cancer immunotherapy treatment with an inhibitor of IRE (inositol-requiring enzyme 1) alpha or XBP1 (X-box binding protein 1), can cause glucose deficiency in tumor microenvironment of all solid cancers. The present invention has been completed by discovering that the sensitivity of cancer cells having nutritional stress to anti-cancer immunotherapy is significantly improved.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for enhancing sensitivity to anti-cancer immunotherapy, comprising an inhibitor of IRE1α or XBP1 as an active ingredient.

However, the problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those having ordinary knowledge in the art from the following description.

According to an embodiment of the present invention, there is provided a pharmaceutical composition for enhancing sensitivity to anti-cancer immunotherapy, comprising an active inhibitor of IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) as an active ingredient. .

According to one side, the composition may be to enhance the anti-cancer immunotherapy sensitivity of cancer cells exposed to hypoglycemia.

According to one side, the activity inhibitor of IRE1α or XBP1 may be STF-083010.

According to one side, the anti-cancer immunotherapy may be T-cell based therapy.

According to one side, the T-cell based therapy may be an immune checkpoint inhibitor or CAR-T cell therapy.

According to one side, the cancer may be one or more cancers selected from the group consisting of lung cancer, breast cancer, pancreatic cancer, and liver cancer.

The pharmaceutical composition for enhancing sensitivity to anti-cancer immunotherapy, comprising the active inhibitor of IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) of the present invention as an active ingredient, when used in combination with anti-cancer immunotherapy, all It is possible to significantly enhance the sensitivity of cancer cells with nutrient stress, such as glucose deficiency in the tumor microenvironment of solid cancer, to anticancer immunotherapy.

The pharmaceutical composition for enhancing the sensitivity of anticancer immunotherapy of the present invention can effectively induce cancer cell death by enhancing the anticancer immunotherapy sensitivity of cancer cells resistant to anticancer immunotherapy to a degree similar to that of non-resistant cancer cells. In addition, since the sensitivity of the cancer cells themselves to the anti-cancer immunotherapy is enhanced, the dose of the anti-cancer immunotherapy agent can be significantly reduced.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be deduced from the configuration of the invention described in the detailed description or claims of the invention.

1 is a lung cancer cell line A549 cell line is a long-term exposure to tumor microenvironments with glucose concentrations of 11mM, 2.7mM, 1.3mM and 0.6mM, labeled with MALT peptide, and the activity of cytotoxic T cells is lowered in low concentration glucose culture conditions. In order to rule out this, the expression level of the active caspase 3 of the A549 cell line was measured after 5 hours co-culture with MALT-specific cytotoxic T cells under normal culture conditions (11 mM).
FIG. 2 shows the expression level of active caspase 3 in the case where STF-083010 is treated for 2 days after treatment with STF-083010 in an A549 cell line exposed to tumor microenvironment for a long period of time with glucose concentrations of 11 mM and 0.6 mM.
Figure 3 is a human lung cancer cell line A549 after culturing in medium with glucose concentrations of 11mM and 0.6mM, Granzyme B kill analysis was performed to compare the expression and cell death rate of active caspase 3 with and without STF-083010 treatment. .
Figure 4 is a human liver cancer cell line HepG2 after culturing in medium with glucose concentrations of 11mM and 0.6mM, Granzyme B kill analysis was performed to compare the expression and cell death rate of active caspase 3 according to the presence or absence of STF-083010 treatment. .
5 is a human pancreatic cancer cell line PANC-1 incubated in a medium having a glucose concentration of 11mM and 0.6mM, and then proceeding to Granzyme B kill analysis to compare the expression and cell death rate of active caspase 3 according to the presence or absence of STF-083010 treatment. Did.
6 is a human breast cancer cell line MDA-MB-231 incubated in a medium having a glucose concentration of 11 mM and 0.6 mM, and then proceeding to Granzyme B killing analysis, the expression of active caspase 3 according to the presence or absence of STF-083010 treatment and the cell death rate It is compared.

Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. However, various changes may be made to the embodiments, and the scope of the patent application right is not limited or limited by these embodiments. It should be understood that all modifications, equivalents, or substitutes for the embodiments are included in the scope of rights.

The terms used in the examples are used for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this specification, the terms "include" or "have" are intended to indicate the presence of features, numbers, steps, actions, components, parts or combinations thereof described herein, one or more other features. It should be understood that the existence or addition possibilities of fields or numbers, steps, operations, components, parts or combinations thereof are not excluded in advance.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person skilled in the art to which the embodiment belongs. Terms such as those defined in a commonly used dictionary should be interpreted as having meanings consistent with meanings in the context of related technologies, and should not be interpreted as ideal or excessively formal meanings unless explicitly defined in the present application. Does not.

In addition, in the description with reference to the accompanying drawings, the same reference numerals are assigned to the same components regardless of reference numerals, and redundant descriptions thereof will be omitted. In describing the embodiments, when it is determined that detailed descriptions of related well-known technologies may unnecessarily obscure the subject matter of the embodiments, detailed descriptions thereof will be omitted.

The present inventors show that the expression level of active caspase 3 is significantly lower than that of cancer cells cultured in 11 mM glucose culture conditions even when cancer cells are exposed to a tumor microenvironment where glucose is maintained at a low concentration for a long period of time, even when anti-cancer immunotherapy is performed. Found something. On the other hand, when treating STF-083010, an inhibitor of IRE1α or XBP1 activity, to cancer cells that are resistant to anti-cancer immunotherapy, the present invention was completed by confirming that the sensitivity of cancer cells to anti-cancer immunotherapy increases to the level of cancer cells under normal culture conditions Did.

Therefore, according to an embodiment of the present invention, a pharmaceutical composition for enhancing sensitivity to anticancer immunotherapy, comprising an active inhibitor of IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) as an active ingredient. Is provided. The IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) activity inhibitor is a substance having a function of inhibiting the expression of the IRE1α protein or the endonuclease activity of IRE1α and the XBP1 protein Any substance having a function of inhibiting the expression of or inhibiting the transcriptional regulatory activity of XBP1 can be used without limitation, and specifically, STF-083010 can be used.

In particular, the pharmaceutical composition of the present invention may be exposed to hypoglycemia to enhance the anti-cancer immunotherapy sensitivity of cancer cells exhibiting resistance to anti-cancer agents. In the present invention, the term "exposed to hypoglycemia" refers to the condition of glucose concentration of 0.6 mM to 1.3 mM, which is clinically close to the tumor microenvironment.

The pharmaceutical composition of the present invention can enhance the sensitivity of cancer cells to anti-cancer immunotherapy by using in combination with anti-cancer immunotherapy. The anti-cancer immunotherapy may be any anti-cancer immunotherapy corresponding to T-cell based therapy, but is not limited thereto. For example, an immune checkpoint inhibitor or CAR-T cell therapy is possible.

In addition, the pharmaceutical composition of the present invention can enhance the anti-cancer immunotherapy sensitivity of all solid cancers, for example, enhance the anti-cancer immunotherapy sensitivity to one or more cancers selected from the group consisting of lung cancer, breast cancer, pancreatic cancer and liver cancer. I can do it.

The pharmaceutical composition of the present invention contains the inhibitor of activity of IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) alone, or is prepared by a method known in the pharmaceutical field for use as a medicament It can be mixed with pharmaceutically acceptable carriers, excipients, diluents, stabilizers, preservatives, etc., and can be prepared and used in formulations such as powders, granules, tablets, capsules, or injections. In addition, the composition may be prepared as a sustained release formulation so that release of the active ingredient occurs slowly, including a base used for sustained release purposes in addition to the active ingredient.

As a pharmaceutically acceptable carrier, for example, a carrier for oral administration or a carrier for parenteral administration may be further included. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, it may contain various drug delivery materials used for oral administration to the peptide preparation. In addition, the carrier for parenteral administration may include water, suitable oil, saline, aqueous glucose and glycol, and may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, etc. in addition to the above components.

Meanwhile, among the pharmaceutically acceptable carriers, components selected from enteric polymers, hydrophobic materials, hydrophilic polymers, etc. may be used as the matrix base used for sustained release purposes. Examples of the enteric polymer include polyvinyl acetate phthalate, hydroxypropyl methyl cellulose phthalate, shellac, cellulose acetate phthalate, cellulose propionate phthalate, (methacrylic acid, methyl methacrylate) copolymer, and poly (methacrylic acid, ethyl acrylic) Rate) Copolymers or the like can be used, and one or more types can be used, preferably hydroxypropyl methylcellulose phthalate.

As the diluent, non-aqueous solvents such as propylene glycol, polyethylene glycol, olive oil, and vegetable oils such as peanut oil, brine (preferably 0.8% brine), and water containing a buffering medium (preferably 0.05M phosphate buffer) And an aqueous solvent such as, but is not limited thereto.

The excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk anhydrous, glycerol, propylene, glycol, water , Ethanol, and the like, but is not limited thereto.

Examples of the stabilizing agent include sorbitol, mannitol, starch, sucrose, dextran, glutamate, glucose, and other carbohydrates or powdered milk, proteins such as animal, vegetable or microbial proteins such as serum albumin and casein. no.

The preservative may include thimerosal, merthiolate, gentamicin, neomycin, nistatin, amphotericin B, tetracycline, penicillin, streptomycin, polymyxin B, and the like, but is not limited thereto.

The pharmaceutical composition of the present invention can be administered in any way to mammals, including humans, and can be administered, for example, orally or parenterally. Parenteral administration methods include intravenous, intramuscular, intraarterial, intramedullary, intrathecal heart, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration. no.

The pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or parenteral administration according to the administration route as described above.

The total effective amount of the pharmaceutical composition of the present invention can be administered to a patient in a single dose, and can be administered in a divided therapy regimen that is administered for a long time in multiple doses. The pharmaceutical composition of the present invention may vary the content of the active ingredient according to the degree of disease. This can be determined in consideration of various factors such as the formulation method, the number of administration routes, and the patient's age, weight, health status, disease symptoms, administration time, and method. In view of this, one of ordinary skill in the art will be able to determine an appropriate effective dose of the composition of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in the formulation, route of administration and method of administration as long as it shows the effect of the present invention.

Hereinafter, the present invention will be described in more detail through examples. The following examples are described for the purpose of illustrating the present invention, and the scope of the present invention is not limited thereto.

Example  1: Sensitivity of anticancer immunotherapy to lung cancer cell lines exposed to low-dose glucose tumor microenvironment for a long time

The lung cancer cell line, A549 cell line, was cultured for 77 days in a medium containing 0.6 mM, 1.3 mM, 2.7 mM, and 11 mM glucose, respectively. The cultured cell line is labeled with MALT peptide, and co-cultured with MALT-specific cytotoxic T cells for 5 hours under normal culture conditions (11 mM) in order to exclude the activity of cytotoxic T cells from being lowered in low-glucose culture conditions, followed by active type The expression level of caspase 3 was measured by FACS.

As a result, as shown in FIG. 1, it was confirmed that the expression level of active caspase 3 in the cell line cultured in culture conditions containing 0.6 mM and 1.3 mM concentration of glucose was lower than that in the 11 mM cultured cell line. Through these results, it can be seen that when the cancer cell line is exposed to a low concentration of glucose environment, the sensitivity of anticancer immunotherapy decreases.

Example  2: XBP1  Increased susceptibility to anticancer immunotherapy following treatment with active inhibitors

The A549 cell line was cultured for 77 days in a medium containing 0.6 mM and 11 mM glucose, respectively. The cultured cell lines were treated with STF-083010 for 2 days, and then cytotoxic T cell-based immunotherapy was performed to measure the expression level of active caspase 3.

As a result, as shown in FIG. 2, when STF-083010 was treated, the expression level of active caspase 3 increased, and cell lines cultured under low concentration (0.6 mM) glucose culture conditions and cell lines cultured under high concentration (11 mM) culture conditions It was confirmed that the expression level of the active caspase 3 was increased to a similar degree.

These results suggest that STF-083010 has the effect of increasing the sensitivity of anti-cancer immunotherapy to cell lines exposed to culture conditions containing low concentrations of glucose.

Example  3: Granzyme  B Death Analysis ( Granzyme  B killing assay)

To reproduce the apoptosis assay of cytotoxic T lymphocytes (CTL), the DOTAP + DOPE complex, a lipid-based delivery system for delivering granzyme B into cells, and granzyme B are mixed to treat cancer cells, and activated caspase The expression of 3 was confirmed by FACs to investigate whether cell death.

Granzyme B killing assays include lung cancer cell line A549 cultured for 77 days in medium containing 0.6 mM and 11 mM glucose, liver cancer cell line HepG2 cultured for 112 days, pancreatic cancer cell line PANC-1 cultured for 91 days, for 70 days The cultured breast cancer cell line MDA-MB-231 was performed. The STF-083010 treatment group was treated with STF-083010 for 2 days in the cell line cultured in a medium containing 0.6 mM and 11 mM glucose, and then granzyme B kill analysis was performed.

As a result, as can be seen in Figures 3 to 6, unlike cell lines cultured in normal culture conditions (11mM), cell death in Granzyme B is inhibited in cell lines cultured in glucose-deficient culture conditions (0.6mM). , In the case of processing STF-083010, it was confirmed that the suppressed apoptosis increased.

Through the above results, it was found that the inhibitor of activity of IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) of the present invention enhances the sensitivity of cancer cells exposed to hypoglycemia to anticancer immunotherapy. Can be.

Therefore, the composition of the present invention can enhance the therapeutic effect when administered in combination with a T-cell based anti-cancer immunotherapeutic agent.

As described above, although the embodiments have been described with limited drawings, those skilled in the art may apply various technical modifications and variations based on the above. For example, even if the described techniques are performed in a different order than the described method, and / or the described components are combined or combined in a different form from the described method, or replaced or replaced by another component or equivalent Appropriate results can be achieved.

Therefore, other implementations, other embodiments, and equivalents to the claims are also within the scope of the following claims.

Claims (6)

IRE1 (inositol-requiring enzyme 1) α or XBP1 (X-box binding protein 1) containing an active inhibitor as an active ingredient, anti-cancer immunotherapy pharmaceutical composition for enhancing sensitivity.
According to claim 1,
The pharmaceutical composition is to enhance the anti-cancer immunotherapy sensitivity of cancer cells exposed to hypoglycemia, the pharmaceutical composition for enhancing the anti-cancer immunotherapy sensitivity.
According to claim 1,
The inhibitor of activity of IRE1α or XBP1 is STF-083010, a pharmaceutical composition for enhancing sensitivity to anticancer immunotherapy.
According to claim 1,
The anti-cancer immunotherapy is a T-cell based therapy, a pharmaceutical composition for enhancing the sensitivity of anticancer immunotherapy.
The method of claim 4,
The T-cell based therapy is an immune checkpoint inhibitor or CAR-T cell therapy, a pharmaceutical composition for enhancing the sensitivity to anticancer immunotherapy.
The method according to any one of claims 1 to 5,
The cancer is one or more cancers selected from the group consisting of lung cancer, breast cancer, pancreatic cancer, and liver cancer.

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