Cancer Immunotherapy Adjuvant

Abstract

The present invention relates to a cancer immunotherapy adjuvant, wherein the adjuvant, when administered in combination with a cancer immunotherapy agent, activates the function of immune factors without causing in vivo side effects, to exhibit the effect of enhancing the kit for anticancer effect of the cancer immunotherapy agent, and thus can be effectively used as a cancer immunotherapy adjuvant.

Description

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a cancer immunotherapy adjuvant.

2. Description of the Related Art

Cancer is a disease characterized by abnormal, localized cell growth that has the potential to spread throughout the body. There are many types of cancer including lung cancer, bladder cancer, prostate cancer, pancreatic cancer, cervical cancer, brain cancer, stomach cancer, colorectal cancer and melanoma. In the past, the most common methods to treat oncological cancer (oncology) were surgery, radiation therapy or chemotherapy. However, it has recently been demonstrated that cancer immunotherapy has many promises as a treatment for oncology.

Cancer immunotherapy is a branch of oncology in which the immune system is used to treat cancer, as opposed to conventional treatment methods in which tumors are directly excised or treated. This therapeutic concept is based on the identification of many proteins on the surface of T cells that act to inhibit the immune function of these cells.

The most fundamental problem in tumor immunity is how to activate the immune system to recognize and eliminate antigens. In this respect, a novel method of genetically engineering tumor cells to secrete specific cytokines has led to major advances in tumor immunity.

The theoretical background of genetically modified tumor vaccines based on immunotherapy is that the host possesses antigens that can recognize tumors as external factors. Human T and B lymphocytes have the ability to discriminate almost infinite antigen differences in the form of antigen receptors through the development process. However, in order to actually succeed in tumor immunity, the following two criteria must be met. First, tumor cells must express novel antigens (peptides) that are not expressed in normal cells. Second, immune cells must be properly activated to recognize these antigens.

The conventional immunotherapy using tumor cells introduced with cytokine genes showed its effectiveness in animal experiments using white mice. Currently, studies are being conducted around the world that when tumor cells introduced with specific cytokine genes are injected into white papers, new tumors can be eradicated and tumor immunity can be acquired in these white mice.

Globally, more than 10 million people are diagnosed with cancer each year, and this number will increase to 15 million new cases annually by 2020. Cancer causes 6 million deaths each year, or 12% of deaths worldwide. There remains a need for methods that can treat cancer. These methods can provide a basis for pharmaceutical compositions useful for the prevention or treatment of cancer in humans and other mammals.

In particular, co-administration for the treatment of cancer is becoming more and more common as the benefit of attacking the disease through multiple means is recognized. Co-administration is useful even when resistance to anticancer drugs is shown. In addition, co-administration has the advantage of reducing the amount of the anticancer agent administered by enhancing the efficacy of the anticancer agent. Through this, it is possible to increase the anticancer efficacy while minimizing the toxicity and side effects on each organ of the body. Although a number of effective combination therapies have been identified over the past few decades; in view of the continuing high annual cancer deaths, there is a continuing need to identify effective therapies for use in anticancer therapies.

Patent reference 1 discloses a pharmaceutical composition for use in co-administration for the prevention or treatment of cancer, comprising a p53 activator and a c-Met inhibitor as active ingredients.

SUMMARY OF THE INVENTION

It is an object of the present invention to provide a cancer immunotherapy adjuvant.

It is another object of the present invention to provide a combination drug for cancer immunotherapy comprising a cancer immunotherapy agent; and a cancer immunotherapy adjuvant.

It is another object of the present invention to provide a pharmaceutical composition for use in enhancing the efficacy of a cancer immunotherapy agent.

It is another object of the present invention to provide a pharmaceutical composition for use in enhancing immunity.

It is another object of the present invention to provide a method for preventing or treating cancer.

It is another object of the present invention to provide a kit for anticancer treatment.

To achieve the above objects, in an aspect of the present invention, the present invention provides a cancer immunotherapy adjuvant comprising a compound represented by formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

(In formula 1,

X is oxygen or sulfur;

represents single bond or double bond;

R₁ is hydrogen, halo, C_1-30 alkyl, C_3-10 cycloalkyl, C_2-30 alkenyl, C_3-10 cycloalkenyl, C_2-15 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-15 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-30 alkoxyalkyl, C_3-30 alkoxyalkyl, C_3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-20 alcohol, C_1-20 alkenol, C_2-30 acyl, C_1-10 amide, C_1-10 amine, C_2-15 ester, sulfate, carboxyl group, C_3-20 carboxyalkyl, C_3-20 carboxyalkenyl, C_3-20 alkylcarboxyl, C_3-20 alkenylcarboxyl, C_3-20 alkylcarboxyalkyl, C_3-20 alkylcarboxyalkenyl, C_3-20 alkenylcarboxyalkyl, C_4-20 alkenylcarboxyalkenyl, C_6-30 aryl, C_6-30 aralkyl, C_6-30 alkaryl, C_3-30 heteroaryl or C_6-30 arylcarbonyl containing nitrogen as a heteroatom;

R₂₁ is C_2-30 alkyl, C_3-10 cycloalkyl, C_2-30 alkenyl, C_3-10 cycloalkenyl, C_2-30 carboxyalkyl, C_2-30 alkylcarboxyl, C_3-30 carboxyalkenyl, C_3-30 alkenylcarboxyl, C_3-30 alkylcarboxyalkyl, C_3-30 alkylcarboxyalkenyl, C_3-30 alkenylcarboxyalkyl, C_4-30 alkenylcarboxyalkenyl, C_2-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-30 alkoxyalkyl, C_3-30 alkoxyalkyl, C_3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-20 alcohol, C_1-20 alkenol, C_2-30 acyl, 0 amide, C_1-10 amine or C_2-15 ester;

R₂₂ is hydrogen, hydroxy, halo or C_1-10 alkyl;

R₂₃ is hydrogen, hydroxyl or C_1-10 alkyl;

R₂₁ may form double bond to the carbon bonded together with R₂₂ and R₂₃;

R₂₃ may form double bond to the carbon bonded together with R₂₁ and R₂₂;

when R₂₁ or R₂₃ forms double bond to the carbon, R₂₂ contains no atoms; and R₃ and R₄ are independently hydrogen or C_1-10 alkyl).

In another aspect of the present invention, the present invention provides a combination drug for cancer immunotherapy comprising a cancer immunotherapy agent; and a cancer immunotherapy adjuvant.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for use in enhancing the efficacy of a cancer immunotherapy agent comprising a compound represented by formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for use in enhancing immunity comprising a compound represented by formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect of the present invention, the present invention provides a method for preventing or treating cancer comprising a step of administering a cancer immunotherapy agent; and a cancer immunotherapy adjuvant to a subject in need thereof.

In another aspect of the present invention, the present invention provides a kit for anticancer treatment comprising a cancer immunotherapy agent; and a cancer immunotherapy adjuvant as active ingredients.

Advantageous Effect

The cancer immunotherapy adjuvant according to the present invention, when administered in combination with a cancer immunotherapy agent, activates the function of immune factors without causing in vivo side effects, to exhibit the effect of enhancing the kit for anticancer effect of the cancer immunotherapy agent, and thus can be effectively used as a cancer immunotherapy adjuvant.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a is a schematic diagram showing the procedure from the preparation of an experimental animal model to the sacrifice of the animal model.

FIG. 1b is a graph showing the changes in tumor size in mice according to the MC38 colorectal cancer cell line injection and drug administration.

FIG. 1c is a graph showing the survival rate of mice according to the MC38 colorectal cancer cell line injection and drug administration.

FIG. 2a is a photograph of a mouse before the excision of the tumor and spleen after the MC38 colorectal cancer cell line injection and drug administration.

FIG. 2b is a photograph of the spleen extracted from the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 2c is a photograph of the tumor extracted from the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 2d is a graph showing the changes in the weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 2e is a graph showing the tumor weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 2f is a graph showing the changes in the spleen weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIGS. 3a to 3c are the results of FACS of the tumor of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 3d is a graph showing the level of immune factors in each drug administration group through the CD45.2+ marker.

FIG. 3e is a graph showing the level of CD4⁺ T cells in each drug administration group.

FIG. 3f is a graph showing the level of CD8⁺ T cells in each drug administration group.

FIG. 3g is a graph showing the level of natural killer cells in each drug administration group.

FIG. 3h is a graph showing the level of regulatory T cells in each drug administration group.

FIG. 4a is a graph showing the proliferative capacity of CD4⁺ T cells in each drug administration group as total %.

FIG. 4b is a graph showing the proliferative capacity of CD8⁺ T cells in each drug administration group as total %.

FIG. 4c is a graph showing the proliferative capacity of natural miller cells in each drug administration group as total %.

FIG. 4d is a graph showing the proliferative capacity of CD4⁺ T cells in each drug administration group as MFI (mean fluorescence intensity).

FIG. 4e is a graph showing the proliferative capacity of CD8⁺ T cells in each drug administration group as MFI (mean fluorescence intensity).

FIG. 4f is a graph showing the proliferative capacity of natural killer cells in each drug administration group as MFI (mean fluorescence intensity).

FIG. 5a is a graph showing the level of CD107a in CD4⁺ T cells in each drug administration group.

FIG. 5b is a graph showing the level of CD107a in CD8⁺ T cells in each drug administration group.

FIG. 6a is a graph showing the level of TNFα in CD4⁺ T cells in each drug administration group.

FIG. 6b is a graph showing the level of TNFα in CD8⁺ T cells in each drug administration group.

FIG. 7a is a graph showing the level of IFNγ in CD4⁺ T cells in each drug administration group.

FIG. 7b is a graph showing the level of IFNγ in CD8⁺ T cells in each drug administration group.

FIGS. 8a and 8b are the results of FACS of the tumor of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 8c is a graph showing the CD107a level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8d is a graph showing the TNFα level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8e is a graph showing the IFNγ level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8f is a graph showing the CD107a level of CD8⁺ T cells in the spleen of each drug administration group.

FIG. 8g is a graph showing the TNFα level of CD8⁺ T cells in the spleen of each drug administration group.

FIG. 8h is a graph showing the IFNγ level of CD8⁺ T cells in the spleen of each drug administration group.

FIG. 9a is a fluorescence photograph showing the expression level of intratumoral adherent junction in each drug administration group.

FIG. 9b is a graph showing the fluorescence density by quantifying the expression level of intratumoral adherent junction in each drug administration group.

FIG. 10a is a fluorescence photograph showing the expression levels of PDL1 and CD3 in each drug administration group.

FIG. 11a is a photograph showing the results of RT-PCR for the expressions of pro-inflammatory cytokines and anti-inflammatory cytokines in each drug administration group.

FIG. 11b is a photograph showing the results of RT-PCR for the expressions of CXCL9, iNOS and Gapdh in each drug administration group.

FIG. 11c is a photograph showing the mRNA expression level graphically through RT-PCR for each group.

FIG. 12a is a schematic diagram showing the procedure according to CD4/8+ T, NK removal, MC38 colorectal cancer cell line injection and drug administration.

FIG. 12b is a diagram confirming that the results according to CD4/8+ T, NK removal, MC38 colorectal cancer cell line injection and drug administration were confirmed through flow cytometry, and that the removal of immune cells proceeded smoothly.

FIG. 12c is a diagram showing the survival rate of the mouse according to CD4/8+ T and NK removal after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the experimental group mouse in which CD8+ T cells were removed had the lowest survival rate.

FIG. 12d is a diagram illustrating the same process as FIG. 12c, showing the growth rate of the tumor according to the CD4/8+ T removal over time after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the tumor growth rate was the highest in the experimental group mouse in which CD8+ T cells were removed.

FIG. 12e is a diagram showing the comparison of the tumor size according to the CD4/8+ T and NK removal in each experimental group after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the tumor size was the largest in the experimental group mouse in which CD8+ T cells were removed.

FIG. 13a is a schematic diagram showing the procedure of the MC38 colorectal cancer cell line injection and long-term drug administration.

FIG. 13b is a graph showing the survival rate of the mouse according to the MC38 colorectal cancer cell line injection and long-term drug administration.

FIG. 13c is a graph showing the tumor size in the mouse according to the MC38 colorectal cancer cell line injection and long-term drug administration.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Hereinafter, the present invention is described in detail.

The embodiments of this invention can be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. It is well understood by those in the art who has the average knowledge on this field that the embodiments of the present invention are given to explain the present invention more precisely. In addition, the “inclusion” of an element throughout the specification does not exclude other elements, but may include other elements, unless specifically stated otherwise.

In an aspect of the present invention, the present invention provides a cancer immunotherapy adjuvant.

Particularly, the present invention provides a cancer immunotherapy adjuvant comprising a compound represented by formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

(In formula 1,

X is oxygen or sulfur;

represents single bond or double bond;

R₁ is hydrogen, halo, C_1-30 alkyl, C_3-10 cycloalkyl, C_2-30 alkenyl, C_3-10 cycloalkenyl, C_2-15 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-15 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-30 alkoxyalkyl, C_3-30 alkoxyalkyl, C_3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-20 alcohol, C_1-20 alkenol, C_2-30 acyl, C_1-10 amide, C_1-10 amine, C_2-15 ester, sulfate, carboxyl group, C_3-20 carboxyalkyl, C_3-20 carboxyalkenyl, C_3-20 alkylcarboxyl, C_3-20 alkenylcarboxyl, C_3-20 alkylcarboxyalkyl, C_3-20 alkylcarboxyalkenyl, C_3-20 alkenylcarboxyalkyl, C_4-20 alkenylcarboxyalkenyl, C_6-30 aryl, C_6-30 aralkyl, C_6-30 alkaryl, C_3-30 heteroaryl or C_6-30 arylcarbonyl containing nitrogen as a heteroatom;

R₂₁ is C_2-30 alkyl, C_3-10 cycloalkyl, C_2-30 alkenyl, C_3-10 cycloalkenyl, C_2-30 carboxyalkyl, C_2-30 alkylcarboxyl, C_3-30 carboxyalkenyl, C_3-30 alkenylcarboxyl, C_3-30 alkylcarboxyalkyl, C_3-30 alkylcarboxyalkenyl, C_3-30 alkenylcarboxyalkyl, C_4-30 alkenylcarboxyalkenyl, C_2-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-30 alkoxyalkyl, C_3-30 alkoxyalkyl, C_3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-20 alcohol, C_1-20 alkenol, C_2-30 acyl, C_1-10 amide, C_1-10 amine or C_2-15 ester;

R₂₂ is hydrogen, hydroxy, halo or C_1-10 alkyl;

R₂₃ is hydrogen, hydroxyl or C_1-10 alkyl;

R₂₁ may form double bond to the carbon bonded together with R₂₂ and R₂₃;

R₂₃ may form double bond to the carbon bonded together with R₂₁ and R₂₂;

when R₂₁ or R₂₃ forms double bond to the carbon, R₂₂ contains no atoms; and

R₃ and R₄ are independently hydrogen or C_1-10 alkyl).

X in formula 1 may be oxygen.

R₁ in formula 1 may be hydrogen, halo, C_1-10 alkyl, C_3-8 cycloalkyl, C_2-10 alkenyl, C_3-8 cycloalkenyl, C_2-8 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-20 alkoxyalkyl, C_3-20 alkoxyalkyl, C_3-8 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-10 alcohol, C_1-10 alkenol, C_2-20 acyl, C_1-10 amide, C_1-5 amine, C_2-15 ester, sulfate, carboxyl group, C_3-20 carboxyalkyl, C_3-20 carboxyalkenyl, C_3-20 alkylcarboxyl, C_3-20 alkenylcarboxyl, C_3-20 alkylcarboxyalkyl, C_3-20 alkylcarboxyalkenyl, C_3-20 alkenylcarboxyalkyl, C_4-20 alkenylcarboxyalkenyl, C_6-20 aryl, C_6-20 aralkyl, C_6-20 alkaryl, C_3-20 heteroaryl or C_6-20 arylcarbonyl containing nitrogen as a heteroatom.

R₁ in formula 1 may be hydrogen, 0 alkyl, C_3-8 cycloalkyl, C_2-10 alkenyl, C_3-8 cycloalkenyl, C_2-8 heterocycloalkyl containing oxygen as a heteroatom, C_3-10 heterocycloalkyl containing oxygen as a heteroatom, C_2-20 alkoxyalkyl, C_3-10 alkoxyalkyl, C_3-8 heterocycloalkenyl containing oxygen as a heteroatom, 0 alcohol, 0 alkenol, C_1-10 amide, C_1-5 amine, C_2-15 ester, sulfate, carboxyl group, C_3-20 carboxyalkyl, C_3-20 carboxyalkenyl, C_3-20 alkylcarboxyl, C_3-20 alkenylcarboxyl, C_3-20 alkylcarboxyalkyl, C_3-20 alkylcarboxyalkenyl, C_3-20 alkenylcarboxyalkyl, C_4-20 alkenylcarboxyalkenyl, C_6-20 aryl, C_6-20 aralkyl, C_6-20 alkaryl, C_3-20 heteroaryl or C_6-20 arylcarbonyl containing nitrogen as a heteroatom.

In R₁ of formula 1, cycloalkyl or heterocycloalkyl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, C_6-20 aryl, C_7-20 arylcarboxyl or a combination thereof; C_3-10 cycloalkenyl or heterocycloalkenyl can be substituted with hydroxy, halo, C_1-5 alkyl, C_2-8 alkylcarboxyl, C_3-8 alkylcarboxylalkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, C_6-20 aryl, C_7-20 arylcarboxyl or a combination thereof; aryl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, nitro, C_2-8 alkylcarboxylamino or a combination thereof; aralkyl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, nitro, C_2-8 alkylcarboxylamino or a combination thereof; alkaryl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, nitro, C_2-8 alkylcarboxylamino or a combination thereof; arylcarbonyl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, nitro, C_2-8 alkylcarboxylamino or a combination thereof; and heteroaryl can be substituted with hydroxy, halo, C_1-5 alkyl, C_1-5 alcohol, C_1-5 alkoxy, C_2-8 alkoxyalkyl, nitro, C_2-8 alkylcarboxylamino or a combination thereof.

R₂₁ in formula 1 may be straight or branched C_2-15 alkyl, C_3-10 cycloalkyl, C_2-15 alkenyl, C_3-10 cycloalkenyl, C_2-15 carboxyalkyl, C_2-15 alkylcarboxyl, C_3-15 carboxyalkenyl, C_2-15 alkenylcarboxyl, C_3-15 alkylcarboxyalkyl, C_3-15 alkylcarboxyalkenyl, C_3-15 alkenylcarboxyalkyl, C_2-30 alkenylcarboxyalkenyl, C_2-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C_2-20 alkoxyalkyl, C_3-30 alkoxyalkyl, C_3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C_1-20 alcohol, C_1-20 alkenol, C_2-30 acyl, amide, C_1-10 amine or C_2-15 ester.

R₂₃ in formula 1 is C_1-5 alkyl or may form double bond to the carbon bonded together with R₂₁ and R₂₂.

in formula 1 may be double bond.

Examples of the compound represented by formula 1 according to the present invention include a compound represented by formula 2 below:

The compound represented by formula 1 of the present invention can be used as a form of a pharmaceutically acceptable salt, in which the salt is preferably acid addition salt formed by pharmaceutically acceptable free acids. The acid addition salt herein can be obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, and phosphorous acid; non-toxic organic acids such as aliphatic mono and dicarboxylate, phenyl-substituted alkanoate, hydroxy alkanoate and alkandioate, aromatic acids, and aliphatic and aromatic sulfonic acids; and organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, and fumaric acid, etc. The pharmaceutically non-toxic salts are exemplified by sulfate, pyrosulfate, bisulfate, sulphite, bisulphite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutylate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, cabacate, fumarate, maliate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutylate, citrate, lactate, β-hydroxybutylate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate.

The acid addition salt according to the present invention can be prepared by the conventional method known to those in the art. For example, the derivative represented by formula 1 is dissolved in an organic solvent such as methanol, ethanol, acetone, methylene chloride, and acetonitrile, to which organic acid or inorganic acid is added to induce precipitation. Then, the precipitate is filtered and dried to give the salt. Or the solvent and the excessive acid are distillated under reduced pressure, and dried to give the salt. Or the precipitate is crystallized in an organic solvent to give the same.

A pharmaceutically acceptable metal salt can be prepared by using a base. Alkali metal or alkali earth metal salt is obtained by the following processes: dissolving the compound in excessive alkali metal hydroxide or alkali earth metal hydroxide solution; filtering non-soluble compound salt; evaporating the remaining solution and drying thereof. At this time, the metal salt is preferably prepared in the pharmaceutically suitable form of sodium, potassium, or calcium salt. And the corresponding silver salt is prepared by the reaction of alkali metal or alkali earth metal salt with proper silver salt (ex; silver nitrate).

In addition, the present invention includes not only the compound represented by formula 1 but also a pharmaceutically acceptable salt thereof, and a solvate, an optical isomer, or a hydrate possibly produced from the same.

The term “hydrate” refers to the compound of the present invention or the salt thereof comprising a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular force. The hydrate of the compound represented by formula 1 of the present invention can include a stoichiometric or non-stoichiometric amount of water that is bound by non-covalent intermolecular force. The hydrate can contain more than 1 equivalent of water, preferably, 1 to 5 equivalents of water. Such a hydrate can be prepared by crystallizing the compound represented by formula 1 of the present invention, the isomer thereof, or the pharmaceutically acceptable salt thereof from water or a solvent containing water.

The term “solvate” refers to the compound of the present invention or the salt thereof comprising a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular force. Preferred solvents therefor include solvents that are volatile, non-toxic, and/or suitable for administration to humans.

The term “isomer” refers to the compound of the present invention or the salt thereof having the same chemical or molecular formula but different structurally or sterically. Such isomers include structural isomers such as tautomers, stereoisomers such as geometric isomers (trans, cis), and optical isomers (enantiomers). All these isomers and mixtures thereof are also included in the scope of the present invention.

In the cancer immunotherapy adjuvant according to the present invention, the compound represented by formula 1 or the pharmaceutically acceptable salt thereof can be administered orally or parenterally in various formulations at the time of clinical administration. More preferably, they can be parenteral formulations. The compound represented by formula 1 or the pharmaceutically acceptable salt thereof can be prepared for oral or parenteral administration by mixing with generally used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactant. Solid formulations for oral administration are tablets, pills, powders, granules and capsules. These solid formulations are prepared by mixing the compound represented by formula 1 or the pharmaceutically acceptable salt thereof of the present invention with one or more suitable excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. Except for the simple excipients, lubricants, for example magnesium stearate, talc, etc, can be used. Liquid formulations for oral administrations are suspensions, solutions, emulsions and syrups, and the above-mentioned formulations can contain various excipients such as wetting agents, sweeteners, aromatics and preservatives in addition to generally used simple diluents such as water and liquid paraffin. Formulations for parenteral administration are sterilized aqueous solutions, water-insoluble excipients, suspensions, and emulsions. Water insoluble excipients and suspensions can contain, in addition to the active compound or compounds, propylene glycol, polyethylene glycol, vegetable oil like olive oil, injectable ester like ethylolate, etc.

The cancer immunotherapy adjuvant comprising the compound represented by formula 1 or the pharmaceutically acceptable salt thereof as an active ingredient can be administered by parenterally and the parenteral administration includes subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

To prepare the compound represented by formula 1 or the pharmaceutically acceptable salt thereof as a formulation for parenteral administration, the compound represented by formula 1 or the pharmaceutically acceptable salt thereof is mixed with a stabilizer or a buffering agent in water to produce solution or suspension, which is then formulated as ampoules or vials. The composition herein can be sterilized and additionally contains preservatives, stabilizers, wettable powders or emulsifiers, salts and/or buffers for the regulation of osmotic pressure, and other therapeutically useful materials, and the composition can be formulated by the conventional mixing, granulating or coating method.

The formulations for oral administration are exemplified by tablets, pills, hard/soft capsules, solutions, suspensions, emulsions, syrups, granules, elixirs, and troches, etc. These formulations can include diluents (for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and/or glycine) and lubricants (for example, silica, talc, stearate and its magnesium or calcium salt, and/or polyethylene glycol) in addition to the active ingredient. Tablets can include binding agents such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrolidone, and if necessary disintegrating agents such as starch, agarose, alginic acid or its sodium salt or azeotropic mixtures and/or absorbents, coloring agents, flavours, and sweeteners can be additionally included thereto.

The cancer immunotherapy adjuvant can enhance the efficacy of the cancer immunotherapy agent, and more specifically, it can enhance the efficacy of the cancer immunotherapy agent by activating immune factors to assist the anticancer activity of the cancer immunotherapy agent. The immune factor can be at least one selected from the group consisting of helper T cells, cytotoxic T cells, natural killer cells (NK cells), and cytokines.

The cancer immunotherapy adjuvant can be administered simultaneously or sequentially with the cancer immunotherapy agent, and when administered sequentially, the cancer immunotherapy adjuvant can be administered after the cancer immunotherapy agent is administered, or the cancer immunotherapy adjuvant can be administered after the cancer immunotherapy agent is administered. However, the administration method is only an example, and the administration method may be changed to enhance the anticancer immune effect. In an embodiment of the present invention, the cancer immunotherapy adjuvant was administered by intravenous injection every day, and the cancer immunotherapy agent was administered by intraperitoneal injection 3 times a week, but not always limited thereto.

The cancer immunotherapy adjuvant can activate one or more immune factors selected from the group consisting of helper T cells, cytotoxic T cells, natural killer cells (NK cells) and cytokines. The cancer immunotherapy adjuvant exhibits the effect of enhancing the anticancer effect of the cancer immunotherapy agent by activating the immune factors.

At this time, the cancer immunotherapy adjuvant can prevent or treat cancer by being administered in combination with the cancer immunotherapy agent.

The cancer can be at least one selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampullar of vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, juvenile brain cancer, juvenile lymphoma, juvenile leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, renal pelvic cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureter cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoma, gastrointestinal stromal cancer, Wilms cancer, breast cancer, triple-negative breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational villous disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, vestibular schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer and thymus cancer.

The cancer immunotherapy adjuvant can be co-administered together with the conventionally known and well-known cancer immunotherapy agent to those skilled in the art without limitation. For example, the cancer immunotherapy adjuvant can be administered in combination with one or more cancer immunotherapy agents selected from the group consisting of anti-PD1, anti-PDL1, anti-CTLA4, anti-LAG3, anti-VISTA, anti-BTLA, anti-TIM3, anti-HVEM, anti-CD27, anti-CD137, anti-OX40, anti-CD28, anti-PDL2, anti-GITR, anti-ICOS, anti-SIRPα, anti-ILT2, anti-ILT3, anti-ILT4, anti-ILT5, anti-EGFR, anti-CD19 and anti-TIGIT, but not always limited thereto.

In another aspect of the present invention, the present invention provides a combination drug for cancer immunotherapy.

Particularly, the present invention provides a combination drug for cancer immunotherapy comprising a cancer immunotherapy agent and a cancer immunotherapy adjuvant.

The specific description of the cancer immunotherapy agent, the cancer immunotherapy adjuvant and the combination drug is the same as the specific description of the cancer immunotherapy adjuvant.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for use in enhancing the efficacy of a cancer immunotherapy agent.

Particularly, the present invention provides a pharmaceutical composition for use in enhancing the efficacy of a cancer immunotherapy agent comprising a compound represented by formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In formula 1, the specific description of R₁ to R₄ and X is the same as the specific description of formula 1 in the cancer immunotherapy adjuvant.

In addition, the specific description of the pharmaceutical composition for use in enhancing the efficacy of a cancer immunotherapy agent is the same as the specific description of the cancer immunotherapy adjuvant.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for use in enhancing immunity.

Particularly, the present invention provides a pharmaceutical composition for use in enhancing immunity comprising a compound represented by formula 1, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In formula 1, the specific description of R₁ to R₄ and X is the same as the specific description of formula 1 in the cancer immunotherapy adjuvant.

In addition, the specific description of the pharmaceutical composition for use in enhancing immunity is the same as the specific description of the cancer immunotherapy adjuvant.

In another aspect of the present invention, the present invention provides a method for preventing or treating cancer comprising a step of administering a cancer immunotherapy agent and a cancer immunotherapy adjuvant to a subject in need thereof.

The cancer immunotherapy adjuvant and the cancer immunotherapy agent can be administered in combination or at different times.

In another aspect of the present invention, the present invention provides a use of a cancer immunotherapy adjuvant and an immunotherapy agent in the prevention or treatment of cancer.

In another aspect of the present invention, the present invention provides a combination therapy for the treatment of cancer comprising a step of administering a cancer immunotherapy adjuvant and a cancer immunotherapy agent to a subject in need thereof.

In another aspect of the present invention, the present invention provides a kit for preventing or treating cancer comprising a cancer immunotherapy agent and a cancer immunotherapy adjuvant as an active ingredient.

Hereinafter, the present invention will be described in detail by the following examples and experimental examples.

However, the following examples and experimental examples are only for illustrating the present invention, and the contents of the present invention are not limited thereto.

Example 1: Preparation of Cancer Immunotherapy Adjuvant (SAC-1004)

The compound SAC-1004 represented by formula 2, the pharmaceutical composition of the present invention for co-administration with a cancer immunotherapy agent, was prepared according to reaction formula 1 below based on Korean Patent Publication No. 10-2011-0047170.

13.4 mg of SAC-1003 was dissolved in 1 mL of tetrahydrofuran, to which 26 mg of tri-O-acetyl-D-glucal (Aldrich) and 0.012 mL of borontrifluoride.diethyl etherate (Aldrich) were added under argon atmosphere, followed by stirring at 0° C. for 10 hours. After raising the temperature of the reaction solution to room temperature, it was diluted by adding 5 mL of diethyl ether, washed with sodium hydrogen carbonate aqueous solution, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography using a mixed eluate of ethyl acetate/hexane (1:10) to give the target compound SAC-1004 (11 mg, 56%): ¹H-NMR (300 MHz, CDCl₃) δ 5.89-5.80 (m, 2H), 5.37-5.27 (m, 2H), 5.17-5.14 (m, 2H), 4.23-4.16 (m, 3H), 3.66 (s, 3H), 3.56 (m, 1H), 2.38-2.28 (m, 4H), 2.17-0.53 (m, 37H).

<Experimental Method>

1. Preparation of Experimental Animal Model

MC-38 colorectal cancer cells (5×10⁵ cells) were injected subcutaneously into 7-week-old C57BL/6 male mice. Seven days after the colorectal cancer cells were injected, the average tumor volume of the subcutaneous MC-38 tumor was about 40 mm³. The injection method of the mouse tumor as described above was equally applied to all four groups below. Afterwards, the experimental group was divided into the vehicle administration control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, and the experiment was performed.

FIG. 1a is a schematic diagram showing the procedure from the preparation of an experimental animal model to the sacrifice of the animal model.

Specifically, the administration was performed as follows. SAC-1004, a vascular leak inhibitor, was dissolved in DMSO in PBS, and 1 mg per kg of mouse body weight was administered daily by intravenous injection. Anti-PD1, an immune checkpoint inhibitor, was administered at the concentration of 200 ng/mouse by intraperitoneal injection 3 times a week along with 200 ng of rat igG2a. As described above, SAC-1004 and anti-PD1 were administered to each group for 7 days, and the tumor and the spleen were extracted on the 8th day.

2. RNA Isolation, cDNA Synthesis and PCR Analysis

Total RNA was isolated from the extracted tumor tissues using Trizol reagent. The concentration of the isolated RNA was determined by measuring the absorbance at 260/280 nm using nano-drop (ND-1000, Thermo scientific), and cDNA was synthesized by including 2 μg of RNA and making the total volume to 20 .

3. Fluorescence-Activated Cell Sorting (FACS) of Mouse Tumor

After separating cells from the tumor tissue using collagenase, a medium was added to the cells, centrifuged, and the precipitated cells were collected and counted. Living cells were stained at 37° C. for about 30 minutes. After staining only living cells, followed by washing with PBS three times. Calibration was conducted through live/dead staining. In addition, FACS was performed using Canto. Through this, the activity ratio of T cells could be compared.

T cells are involved in adaptive immune response and are divided into helper T cells (CD4⁺ T cells), cytotoxic T cells (CD8⁺ T cells), and regulatory T cells (Treg cells) derived from CD4⁺ T cells but suppressing cytotoxic T cells. In addition, there are natural killer cells (NK cells) with cytotoxic properties involved in innate immune response, so the levels of these cells in each group were measured.

CD4 antibody was used as a marker for measuring CD4⁺ T cells, which are helper T cells, and CD8 antibody was used as a marker for measuring CD8⁺ T cells, which are cytotoxic T cells. In addition, CD25 antibody and Foxp³ antibody were used to measure regulatory T cells, and NK1.1 antibody was used to measure natural killer cells.

4. IHC Staining

The tumor tissue was immediately taken out and then 0/N incubated in 4% PFA (stored at 4° C.). After the tumor tissue was submerged sequentially in 15% to 30% sucrose to sink, OCT sampling was performed on dry ice. 20 μm sections were stained with each primary antibody and secondary antibody.

5. Analysis of T Cell Function

The function of T cells is mainly measured in two ways. One method is cytokine production capacity analysis, which measures the secretion of IFNγ or TNFα, which are pro-inflammatory cytokines, from T cells. Another method is a method of measuring cytotoxicity, in which tumor cells are directly ligated and killed by inserting perforin or granzyme B into the tumor cells.

Experimental Example 1: Analysis of Tumor Size Change in Mice According to MC38 Colorectal Cancer Cell Line Injection and Drug Administration

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the tumor size changes according to the drug administration for 7 days as described above in the experimental animal model were analyzed.

FIG. 1b is a graph showing the changes in tumor size in mice according to the MC38 colorectal cancer cell line injection and drug administration.

FIG. 2a is a photograph of a mouse before the excision of the tumor and spleen after the MC38 colorectal cancer cell line injection and drug administration.

FIG. 2c is a photograph of the tumor extracted from the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

Particularly, the tumor size in the anti-PD1 single administration group and the SAC-1004 single administration group was reduced compared to that in the control group, and the tumor size in the SAC-1004 and anti-PD1 co-administration group was decreased compared to those in the single administration groups. The results were statistically proved by two-way ANOVA.

The above results demonstrate that the preventive or therapeutic effect on cancer was significantly increased when SAC-1004 and the immunotherapy agent were administered in combination rather than when administered alone.

Experimental Example 2: Analysis of Survival Rate of Mice According to MC38 Colorectal Cancer Cell Line Injection and Drug Administration

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the survival rate of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed.

FIG. 1c is a graph showing the survival rate of mice according to the MC38 colorectal cancer cell line injection and drug administration.

Particularly, the survival rate of mice according to the drug administration in the anti-PD1 single administration group was higher than that in the control group or the SAC-1004 single administration group, and the survival rate of mice in the SAC-1004 and anti-PD1 co-administration group was higher than that in the anti-PD1 single administration group. The results were statistically proved by two-way ANOVA.

The above results demonstrate that the preventive or therapeutic effect on cancer was significantly increased when SAC-1004 and the immunotherapy agent were administered in combination rather than when administered alone.

Experimental Example 3: Analysis of Weight Changes in Mice According to MC38 Colorectal Cancer Cell Line Injection and Drug Administration

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the weight changes in mice according to the drug administration for 7 days as described above in the experimental animal model were analyzed.

FIG. 2d is a graph showing the changes in the weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

Particularly, similar body weights were measured in all groups, indicating that no abnormalities were observed in mice even after long-term administration of the drug.

The above results demonstrate that the pharmaceutical composition for use in co-administration according to the present invention did not cause side effects in vivo even when administered alone or in combination, and is a material with safety secured.

Experimental Example 4: Comparison of Spleen Size and Weight of Mice Injected with MC38 Colorectal Cancer Cell Line and Administered with Drugs

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the spleen size of mice treated with the drug for 7 days as described above in the experimental animal model was analyzed.

FIG. 2b is a photograph of the spleen extracted from the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 2f is a graph showing the changes in the spleen weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

Particularly, the spleen weight of the SAC-1004 single administration group mice was higher than that of the control group mice, the spleen weight of the anti-PD1 single administration group mice was higher than that of the SAC-1004 single administration group mice, and the spleen weight of the SAC-1004 and anti-PD1 co-administration group mice was higher than that of the anti-PD1 single administration group mice. However, this was not statistically significant when verified by one-way ANOVA.

The above results demonstrate that the pharmaceutical composition for use in co-administration according to the present invention did not cause side effects in vivo even when administered alone or in combination, and is a material with safety secured.

Experimental Example 5: Comparison of Tumor Weight of Mice Injected with MC38 Colorectal Cancer Cell Line and Administered with Drugs

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the tumor weight of mice treated with the drug for 7 days as described above in the experimental animal model was analyzed.

FIG. 2e is a graph showing the tumor weight of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

Particularly, the tumor weight of the anti-PD1 single administration group mice was lower than that of the control group mice, the tumor weight of the SAC-1004 single administration group mice was lower than that of the anti-PD1 single administration group mice, and the tumor weight of the SAC-1004 and anti-PD1 co-administration group mice was lower than that of the SAC-1004 single administration group mice. The results were statistically proved by one-way ANOVA.

The above results demonstrate that the preventive or therapeutic effect on cancer was significantly increased when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone.

Experimental Example 6: FACS Analysis of Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, FACS (fluorescence activated cell sorting) analysis was performed on the tumors of mice treated with the drug for 7 days as described above in the experimental animal model, and the activity ratio of T cells in each drug-administered group was compared.

CD4 antibody was used to measure CD4⁺ T cells, which are helper T cells, and CD8 antibody was used to measure CD8⁺ T cells, which are cytotoxic T cells. In addition, CD25 antibody and Foxp³ antibody were used to measure regulatory T cells, and NK1.1 antibody was used to measure natural killer cells.

FIGS. 3a to 3c are the results of FACS of the tumor of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 3d is a graph showing the level of immune factors in each drug administration group through the CD45.2+ marker.

FIG. 3e is a graph showing the level of CD4⁺ T cells in each drug administration group.

FIG. 3f is a graph showing the level of CD8⁺ T cells in each drug administration group.

FIG. 3g is a graph showing the level of natural killer cells in each drug administration group.

FIG. 3h is a graph showing the level of regulatory T cells in each drug administration group.

Particularly, in the SAC-1004 and anti-PD1 co-administration group, the levels of cytotoxic T cells as well as helper T cells were significantly high, the level of natural killer cells was also high, but the level of regulatory T cells that suppressed cytotoxic T cells was decreased. The results were statistically proved by one-way ANOVA.

Therefore, it was confirmed that the effect of activating T cells is greater when SAC-1004 and the cancer immunotherapy agent are administered in combination than when administered alone.

The above results indicate that the effect of activating T cells, an immune-related factor, was superior when SAC-1004 and the cancer immunotherapy agent were administered in combination than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

Experimental Example 7: Analysis of Proliferative Capacity of Immune Factors in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the proliferative capacity of immune factors in the tumors of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed. To measure the proliferative capacity, Ki67, a proliferation marker, was used.

FIG. 4a is a graph showing the proliferative capacity of CD4⁺ T cells in each drug administration group as total %.

FIG. 4b is a graph showing the proliferative capacity of CD8⁺ T cells in each drug administration group as total %.

FIG. 4c is a graph showing the proliferative capacity of natural miller cells in each drug administration group as total %.

FIG. 4d is a graph showing the proliferative capacity of CD4⁺ T cells in each drug administration group as MFI (mean fluorescence intensity).

FIG. 4e is a graph showing the proliferative capacity of CD8⁺ T cells in each drug administration group as MFI (mean fluorescence intensity).

FIG. 4f is a graph showing the proliferative capacity of natural killer cells in each drug administration group as MFI (mean fluorescence intensity).

Particularly, the proliferative capacity of helper T cells, cytotoxic T cells, and natural killer cells was higher in the SAC-1004 and anti-PD1 co-administration group than in the other groups.

The above results indicate that when SAC-1004 and the cancer immunotherapy agent were administered in combination, the effect of enhancing the proliferative capacity of the immune-related factors was superior to when SAC-1004 and the cancer immunotherapy agent were administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

Experimental Example 8: Analysis of T Cell Cytotoxicity in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the T cell cytotoxicity in the tumors of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed.

To measure the cytotoxicity, CD107a, which is involved in the degranulation of natural killer cells and the activation of CD8⁺ T cells, was used as a marker, and the peptide and PMA/ionomycin were treated to activate T cells. In the group treated with the peptide, the peptide expressed in the tumor was added in vitro. In the group treated with PMA/ionomycin, all T cells were activated by stimulating the T cell calcium signal.

FIG. 5a is a graph showing the level of CD107a in CD4⁺ T cells in each drug administration group.

FIG. 5b is a graph showing the level of CD107a in CD8⁺ T cells in each drug administration group.

Particularly, in the group not treated with the peptide and PMA/ionomycin, the level of CD107a in T cells was lower, and the level of CD107a in T cells in the group treated with PMA/ionomycin was higher than that in the group treated with the peptide. In particular, when PMA/ionomycin was treated, the level of CD107a in CD8⁺ T cells was high in the SAC-1004 and anti-PD1 co-administration group.

The above results, although relatively increased activation in CD8+ T cells, were not significant in all groups. The cytotoxic effect of activating T cells to kill cancer cells was superior when SAC-1004 and the cancer immunotherapy agent were administered in combination to when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect by co-administration with the cancer immunotherapy agent.

Experimental Example 9: Analysis of Cytokine Production Capacity of T Cells in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the cytokine production capacity of T cells in the tumors of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed.

(9-1)

In order to measure the cytokine production capacity, the level of pro-inflammatory cytokines can be compared, and TNFα, a pro-inflammatory cytokine marker, can be used. As in Experimental Example 8, the level of TNFα was measured after treatment with the peptide and PMA/ionomycin to activate T cells.

FIG. 6a is a graph showing the level of TNFα in CD4⁺ T cells in each drug administration group.

FIG. 6b is a graph showing the level of TNFα in CD8⁺ T cells in each drug administration group.

Particularly, when the peptide and PMA/ionomycin were not treated, the difference in the level of TNFα was not significant in each drug administration group, but when the peptide and PMA/ionomycin were treated, the level of TNFα was significantly increased in SAC-1004 and anti-PD1 co-administration group.

The above results indicate that the effect of activating T cells, an immune-related factor, was superior when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

(9-2)

IFNγ is also a proinflammatory cytokine marker. Similarly, the level of IFNγ was measured after treatment with the peptide and PMA/ionomycin to activate T cells.

FIG. 7a is a graph showing the level of IFNγ in CD4⁺ T cells in each drug administration group.

FIG. 7b is a graph showing the level of IFNγ in CD8⁺ T cells in each drug administration group.

Particularly, when the peptide and PMA/ionomycin were not treated, the difference in the level of IFNγ was not significant in each drug administration group, but when the peptide and PMA/ionomycin were treated, the level of IFNγ was significantly increased in SAC-1004 and anti-PD1 co-administration group.

The above results indicate that the effect of activating T cells, an immune-related factor, was superior when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

Experimental Example 10: Analysis of Cytokine Production Capacity of T Cells in Mouse Spleen

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, FACS analysis, cytotoxicity analysis, and cytokine production capacity analysis were performed using CD107a, TNFα and IFNγ markers to compare the functions of T cells in the spleen of mice treated with the drug for 7 days as described above in the experimental animal model. In addition, as in Experimental Examples 8 and 9, the peptide and PMA/ionomycin were treated to activate T cells.

FIGS. 8a and 8b are the results of FACS of the tumor of the mouse injected with MC38 colorectal cancer cell line and administered with drugs.

FIG. 8c is a graph showing the CD107a level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8d is a graph showing the TNFα level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8e is a graph showing the IFNγ level of CD4⁺ T cells in the spleen of each drug administration group.

FIG. 8f is a graph showing the CD107a level of CD8⁺ T cells in the spleen of each drug administration group.

FIG. 8g is a graph showing the TNFα level of CD8⁺ T cells in the spleen of each drug administration group.

FIG. 8h is a graph showing the IFNγ level of CD8⁺ T cells in the spleen of each drug administration group.

Particularly, when T cells were activated by treating CD4⁺ T cells and CD8⁺ T cells in the spleen with PMA/ionomycin, the levels of CD107a, TNFα and IFNγ were increased in the SAC-1004 and anti-PD1 co-administration group.

The above results indicate that the cytotoxic effect of killing cancer cells was superior and the effect of activating T cells, an immune-related factor, was also superior when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

Experimental Example 11: Analysis of Adherent Junction Expression in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the expression of adherent junction in the tumor of the mouse treated with the drug as described above in the experimental animal model was analyzed except that the drug was administered for 14 days by daily administration of SAC-1004 for 7 days and administration of anti-PD1 3 times a week.

FIG. 9a is a fluorescence photograph showing the expression level of intratumoral adherent junction in each drug administration group.

FIG. 9b is a graph showing the fluorescence density by quantifying the expression level of intratumoral adherent junction in each drug administration group.

Particularly, FIG. 9a shows the results of IHC staining, DAPI was used to stain the nucleus, CD31 was used to stain blood vessels, and VE-cadherin (adhesion protein) was used to view adherent junction. At this time, if VE-cadherin is well expressed, drug delivery is good and the anticancer effect is increased, and the adhesion between endothelial cells is good, blood vessels are stabilized, and T cells are well infiltrated into the tumor.

As shown in FIGS. 9a and 9b, the ratio of intratumoral VE-cadherin was increased in the SAC-1004 single administration group than that in the control group and the anti-PD1 single administration group, and intratumoral VE-cadherin was more expressed in the SAC-1004 and anti-PD1 co-administration group than in the SAC-1004 single administration group.

The above results indicate that the cancer immunotherapy agent was well delivered, T cells penetrated well into the tumor, and the anticancer effect was increased when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent.

Experimental Example 12: Analysis of PDL1 Expression in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the expression of PDL1 (programmed death-ligand 1) in the tumors of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed. PDL1 is a ligand expressed in tumors, and CD3 is used as a T cell marker.

FIG. 10a is a fluorescence photograph showing the expression levels of PDL1 and CD3 in each drug administration group.

Particularly, FIG. 10a shows the results of IHC staining, and the increase in PDL1 means that anti-PD1 was well delivered and immune activation occurred. The expression levels of PDL1 and CD3 in the SAC-1004 and anti-PD1 co-administration group were higher than those in the anti-PD1 single administration group.

The above results indicate that the cancer immunotherapy agent was well delivered, immune activation occurred well, and the anticancer effect was increased when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent.

Experimental Example 13: Analysis of Cytokine Expression Level in Mouse Tumor

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the expression level of cytokines in the tumors of mice according to the drug administration for 7 days as described above in the experimental animal model was analyzed. As cytokine markers, pro-inflammatory cytokines and anti-inflammatory cytokines were used, and RT-PCR was performed.

FIG. 11a is a photograph showing the results of RT-PCR for the expressions of pro-inflammatory cytokines and anti-inflammatory cytokines in each drug administration group.

FIG. 11b is a photograph showing the results of RT-PCR for the expressions of CXCL9, iNOS and Gapdh in each drug administration group.

FIG. 11c is a photograph showing the mRNA expression level graphically through RT-PCR for each group.

Particularly, among pro-inflammatory cytokines, IFNγ was increased in the group administered with anti-PD1, and it was more increased in the SAC-1004 and anti-PD1 co-administration group than in the group administered with anti-PD1.

The above results indicate that the effect of activating T cells, an immune-related factor, was superior when SAC-1004 and the cancer immunotherapy agent were administered in combination rather than when administered alone. From the above results, it was confirmed that the pharmaceutical composition for use in co-administration of the present invention can enhance the anticancer effect of the cancer immunotherapy agent by activating immune factors.

Experimental Example 14: Analysis of Survival Rate of Mice According to Drug Administration after Depletion of CD4/8⁺ T and NK Cells at the Time of Growth of MC38 Colorectal Cancer Cell Line and Drug Injection

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the survival rate of mice according to the drug administration for 7 days as described above in the experimental animal model after depletion of CD4/8⁺ T and NK cells was analyzed.

FIG. 12a is a schematic diagram showing the procedure according to CD4/8+ T, NK removal, MC38 colorectal cancer cell line injection and drug administration.

FIG. 12b is a diagram confirming that the results according to CD4/8+ T, NK removal, MC38 colorectal cancer cell line injection and drug administration were confirmed through flow cytometry, and that the removal of immune cells proceeded smoothly.

FIG. 12c is a diagram showing the survival rate of the mouse according to CD4/8+ T and NK removal after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the experimental group mouse in which CD8+ T cells were removed had the lowest survival rate.

FIG. 12d is a diagram illustrating the same process as FIG. 12c, showing the growth rate of the tumor according to the CD4/8+ T removal over time after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the tumor growth rate was the highest in the experimental group mouse in which CD8+ T cells were removed.

FIG. 12e is a diagram showing the comparison of the tumor size according to the CD4/8+ T and NK removal in each experimental group after the MC38 colorectal cancer cell line injection and drug administration, and confirming that the tumor size was the largest in the experimental group mouse in which CD8+ T cells were removed.

The above results indicate that the removal of CD8⁺ T cells showed the greatest tumor growth rate, and that the experimental group in which NK cells and CD4⁺ T cells were removed showed a great tumor growth rate. This suggests that the co-administration of SAC-1004 and anti-PD1 is a CD8⁺ T cell-dependently induced process.

Experimental Example 15: Analysis of Results According to Long-Term Drug Administration

In a total of 4 groups consisting of control group, SAC-1004 and anti-PD1 co-administration group, SAC-1004 single administration group, and anti-PD1 single administration group, the survival rate was analyzed after long-term drug injection.

FIG. 13a is a schematic diagram showing the procedure of the MC38 colorectal cancer cell line injection and long-term drug administration.

FIG. 13b is a graph showing the survival rate of the mouse according to the MC38 colorectal cancer cell line injection and long-term drug administration.

FIG. 13c is a graph showing the tumor size in the mouse according to the MC38 colorectal cancer cell line injection and long-term drug administration.

The same results were consistently shown even after long-term drug treatment. Therefore, the reliability of the above experimental procedure was confirmed, and it was proved that there was no drug toxicity despite long-term drug administration.

Manufacturing Example 1: Preparation of Powders

Compound represented by formula 1 2 g
Lactose                          1 g

Powders were prepared by mixing all the above components, which were filled in airtight packs according to the conventional method for preparing powders.

Manufacturing Example 2: Preparation of Tablets

Compound represented by formula 1 100 mg
Corn starch                      100 mg
Lactose                          100 mg
Magnesium stearate               2 mg

Tablets were prepared by mixing all the above components by the conventional method for preparing tablets.

Manufacturing Example 3: Preparation of Capsules

Compound represented by formula 1 100 mg
Corn starch                      100 mg
Lactose                          100 mg
Magnesium stearate               2 mg

Capsules were prepared by mixing all the above components, which were filled in gelatin capsules according to the conventional method for preparing capsules.

Manufacturing Example 4: Preparation of Injectable Solutions

Compound represented by formula 1 100 mg
Mannitol                         180 mg
Na2HPO4•2H2O                     26 mg
Distilled water                  2974 mg

Injectable solutions were prepared by mixing all the above components by the conventional method for preparing injectable solutions.

Manufacturing Example 5: Preparation of Ointments

Compound represented by formula 1 5 g
Cetyl palm itate                 20 g
Cetanol                          40 g
Stearyl alcohol                  40 g
Isopropyl myristate              80 g
Polysorbate                      60 g
Propyl p-hydroxybenzoate         1 g
Methyl p-hydroxybenzoate         1 g

Phosphoric Acid and Purified Water Proper Amount

Ointments were prepared by mixing all the above components by the conventional method for preparing ointments.

Claims

1-21. (canceled)

22. A combination drug comprising: a cancer immunotherapy agent; anda compound represented by formula 1 below, an isomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:

23. The combination drug according to claim 22, wherein the X in formula 1 is oxygen.

24. The combination drug according to claim 22, wherein the R1 in formula 1 is hydrogen, halo, C1-10 alkyl, C3-8 cycloalkyl, C2-10 alkenyl, C3-8 cycloalkenyl, C2-8 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C2-20 alkoxyalkyl, C3-20 alkoxyalkyl, C3-8 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C1-10 alcohol, C1-10 alkenol, C2-20 acyl, C1-10 amide, C1-5 amine, C2-15 ester, sulfate, carboxyl group, C3-20 carboxyalkyl, C3-20 carboxyalkenyl, C3-20 alkylcarboxyl, C3-20 alkenylcarboxyl, C3-20 alkylcarboxyalkyl, C3-20 alkylcarboxyalkenyl, C3-20 alkenylcarboxyalkyl, C4-20 alkenylcarboxyalkenyl, C6-20 aryl, C6-20 aralkyl, C6-20 alkaryl, C3-20 heteroaryl or C6-20 arylcarbonyl containing nitrogen as a heteroatom.

25. The combination drug according to claim 24, wherein the R1 in formula 1 is hydrogen, C1-10 alkyl, C3-8 cycloalkyl, C2-10 alkenyl, C3-8 cycloalkenyl, C2-8 heterocycloalkyl containing oxygen as a heteroatom, C3-10 heterocycloalkyl containing oxygen as a heteroatom, C2-20 alkoxyalkyl, C3-10 alkoxyalkyl, C3-8 heterocycloalkenyl containing oxygen as a heteroatom, C1-10 alcohol, C1-10 alkenol, C1-10 amide, C1-5 amine, C2-15 ester, sulfate, carboxyl group, C3-20 carboxyalkyl, C3-20 carboxyalkenyl, C3-20 alkylcarboxyl, C3-20 alkenylcarboxyl, C3-20 alkylcarboxyalkyl, C3-20 alkylcarboxyalkenyl, C3-20 alkenylcarboxyalkyl, C4-20 alkenylcarboxyalkenyl, C6-20 aryl, C6-20 aralkyl, C6-20 alkaryl, C3-20 heteroaryl or C6-20 arylcarbonyl containing nitrogen as a heteroatom.

26. The combination drug according to claim 22, wherein the cycloalkyl or heterocycloalkyl in R1 of formula 1 can be substituted with hydroxy, halo, C1-5 alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, C6-20 aryl, C7-20 arylcarboxyl or a combination thereof; C3-10 cycloalkenyl or heterocycloalkenyl can be substituted with hydroxy, halo, C1-5 alkyl, C2-8 alkylcarboxyl, C3-8 alkylcarboxylalkyl, C1-5 alcohol, C1-s alkoxy, C2-8 alkoxyalkyl, C6-20 aryl, C7-20 arylcarboxyl or a combination thereof; aryl can be substituted with hydroxy, halo, C1-s alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, nitro, C2-8 alkylcarboxylamino or a combination thereof; aralkyl can be substituted with hydroxy, halo, C1-5 alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, nitro, C2-8 alkylcarboxylamino or a combination thereof; alkaryl can be substituted with hydroxy, halo, C1-5 alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, nitro, C2-8 alkylcarboxylamino or a combination thereof; arylcarbonyl can be substituted with hydroxy, halo, C1-5 alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, nitro, C2-8 alkylcarboxylamino or a combination thereof; and heteroaryl can be substituted with hydroxy, halo, C1-5 alkyl, C1-5 alcohol, C1-5 alkoxy, C2-8 alkoxyalkyl, nitro, C2-8 alkylcarboxylamino or a combination thereof.

27. The combination drug according to claim 22, wherein the R21 in formula 1 is straight or branched C2-15 alkyl, C3-10 cycloalkyl, C2-15 alkenyl, C3-10 cycloalkenyl, C2-15 carboxyalkyl, C2-15 alkylcarboxyl, C3-15 carboxyalkenyl, C2-15 alkenylcarboxyl, C3-15 alkylcarboxyalkyl, C3-15 alkylcarboxyalkenyl, C3-15 alkenylcarboxyalkyl, C2-30 alkenylcarboxyalkenyl, C2-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C3-10 heterocycloalkyl containing oxygen, sulfur or nitrogen as a heteroatom, C2-20 alkoxyalkyl, C3-30 alkoxyalkyl, C3-10 heterocycloalkenyl containing oxygen, sulfur or nitrogen as a heteroatom, C1-20 alcohol, C1-20 alkenol, C2-30 acyl, C1-10 amide, C1-10 amine or C2-15 ester.

28. The combination drug according to claim 22, wherein the R23 in formula 1 is C1-5 alkyl or forms double bond to the carbon bonded together with R21 and R22.

29. The combination drug according to claim 22, wherein the in formula 1 is double bond.

30. The combination drug according to claim 22, wherein the compound of formula 1 is a compound represented by formula 2 below:

31. The combination drug according to claim 22, wherein the compound enhances the efficacy of a cancer immunotherapy agent.

32. The combination drug according to claim 22, wherein the compound exhibits an immune enhancing effect.

33. The combination drug according to claim 22, wherein the compound can be administered simultaneously or sequentially with a cancer immunotherapy agent.

34. The combination drug according to claim 22, wherein the compound activates at least one immune factor selected from the group consisting of helper T cells, cytotoxic T cells, natural killer cells (NK cells) and cytokines.

35. The combination drug according to claim 22, wherein the compound prevents or treats at least one cancer selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampullar of vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, nasal cavity cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, juvenile brain cancer, juvenile lymphoma, juvenile leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, renal pelvic cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureter cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoma, gastrointestinal stromal cancer, Wilms cancer, breast cancer, triple-negative breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational villous disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, vestibular schwannoma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer and thymus cancer.

36. The combination drug according to claim 22, wherein the compound is co-administered together with at least one cancer immunotherapy agent selected from the group consisting of anti-PD1, anti-PDL1, anti-CTLA4, anti-LAG3, anti-VISTA, anti-BTLA, anti-TIM3, anti-HVEM, anti-CD27, anti-CD137, anti-OX40, anti-CD28, anti-PDL2, anti-GITR, anti-ICOS, anti-SIRPα, anti-ILT2, anti-ILT3, anti-ILT4, anti-ILT5, anti-EGFR, anti-CD19 and anti-TIGIT.

37. The combination drug according to claim 22, wherein the combination drug is a combination drug for cancer immunotherapy.

38. A method for preventing or treating cancer comprising a step of administering the combination drug of claim 22 to a subject in need thereof.

39. A kit for anticancer treatment comprising a cancer immunotherapy agent; and the compound of claim 22 as active ingredients.