KR20220009270A - Endothelin receptor antagonist- encapsulated nanoparticle and pharmaceutical composition for preventing or treating cancer comprising the same as an active ingredient - Google Patents

Abstract

The present invention relates to nanoparticles encapsulated with an endothelin receptor antagonist, a therapeutic use thereof for cancer, and a use for enhancing an anticancer effect of an anticancer agent. The nanoparticles of the present invention accumulate in cancer tissues and release endothelin receptor antagonists, and simultaneously act on tumor-associated fibroblasts, endothelial cells, and cancer cells that play a major role in the tumor microenvironment, thereby controlling the microenvironment within the tumor through inhibition of a fibrosis process and inhibition of an extracellular matrix to significantly improve a response rate and a therapeutic effect of anticancer drugs such as immune checkpoint inhibitors.

Description

Endothelin receptor antagonist- encapsulated nanoparticle and pharmaceutical composition for preventing or treating cancer comprising the same as an active ingredient

The present invention relates to nanoparticles encapsulated with an endothelin receptor antagonist, and a therapeutic use thereof for cancer and a use for enhancing the anticancer effect of an anticancer agent.

In recent years, the paradigm for anticancer treatment is changing from chemotherapy to cancer immunotherapy, in which immune cells attack cancer cells by activating the immune system in the body. It has the advantage of showing long-term therapeutic effects. Immune anticancer drugs can be broadly classified into immune checkpoint inhibitors, immune cell therapeutics, therapeutic antibodies, and anticancer vaccines. It can also be administered to patients, so it is attracting attention in clinical practice.

Immunochemotherapy or anticancer immunotherapy using an immune checkpoint inhibitor is an immune checkpoint molecule such as PD-1, PD-L1, CTLA-4 expressed as an immune evasion mechanism for the survival of cancer cells with the corresponding antibody (anti-PD-1 antibody). , anti-PD-L1 antibody, etc.) is used to block immune response evasion signals, thereby activating T cells to attack cancer cells.

However, although survival rate and therapeutic effect were greatly improved with the advent of checkpoint inhibitors, there was a limitation in that only a small number of patients (15-30%) showed anti-cancer effects against anti-PD-L1/PD-1 antibodies. T cells were sensitized and activated in patients not responsive to checkpoint inhibitors, but defects were observed at the stage of T cell invasion into the tumor. In particular, even if immune cells were present and activated, when T cells did not pass through the parenchyma of the tumor and stayed outside the tumor, the clinical response rate to the anti-PD-L1/PD-1 antibody was low. The migration of T cells is affected by the structure of the tumor microenvironment (TME), and the higher the density of the tumor stroma, the less the number of T cells distributed in the cancer tissue is observed, and the reactivity to anticancer immunotherapy is also low. has been reported Therefore, in order to increase the response rate to the immune checkpoint inhibitor, there is a need for a strategy that can control the tumor microenvironment so that activated T cells can effectively move into the tumor.

Republic of Korea Patent Registration No. 10-1943230

Accordingly, the present inventors control the components of the tumor microenvironment that inhibit the penetration of T cells or anticancer agents into the tumor, and also maximize the cancer treatment effect of anticancer agents including immune checkpoint inhibitors by suppressing the exosome secretion of cancer cells. The present invention was completed by preparing nanoparticles capable of controlling the microenvironment within the tumor.

Accordingly, it is an object of the present invention to provide nanoparticles encapsulating an endothelin receptor antagonist therein.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising the nanoparticles as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for enhancing the anticancer effect of an anticancer agent comprising the nanoparticles as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for inhibiting cancer metastasis comprising the nanoparticles as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned tasks, and other tasks not mentioned can be clearly understood by those of ordinary skill in the art to which the present invention belongs from the following description. will be.

In order to achieve the object of the present invention, the present invention contains a biocompatible amphiphilic polymer comprising a hydrophilic polymer and a hydrophobic polymer conjugated to the hydrophilic polymer, and an endothelin receptor antagonist is encapsulated therein, characterized in that nanoparticles are provided.

In one embodiment of the invention, the endothelin receptor antagonist may _{exhibit antagonistic action against both the endothelin A receptor (ET A} ) and the endothelin B receptor (ET _B ), but is not limited thereto.

In another embodiment of the present invention, the endothelin receptor antagonist is macitentan, bosentan, ambrisentan, sulfisoxazole, BQ-123, BQ-788, zibo It may be selected from the group consisting of zibotentan, sitaxentan, atrasentan, tezosentan, and A192621, but is not limited thereto.

In another embodiment of the present invention, the hydrophilic polymer is carboxymethyl dextran, chitosan, gelatin, collagen, mannan, dextran sulfate , α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hyaluronic acid, alginate, glycogen, amylose, beta-glucan , hydroxyethyl cellulose, carboxymethyl cellulose, fucoidan, chondroitin, and a polysaccharide polymer selected from the group consisting of combinations thereof, but is not limited thereto.

In another embodiment of the present invention, the hydrophilic polymer is polyethylene glycol, polyethylene oxide, polyoxazoline, poly(N-vinylpyrrolidone), polyvinyl alcohol, polyhydroxyethyl methacrylate, polyserine, It may be selected from the group consisting of polythreonine, polytyrosine, polylysine, polyarginine, polyhistidine, polyaspartic acid, polyglutamic acid, and combinations thereof, but is not limited thereto.

In another embodiment of the present invention, the hydrophobic polymer is a polylactic acid-glycolic acid copolymer (Poly (lactic-co-glycolic acid), polyphosphazene, polylactide, polycaprolactone, polyanhydride, It may be selected from the group consisting of polymalic acid, polyalkylcyanoacrylate, polyhydrooxybutylate, polycarbonate, polyorthoester, hydrophobic polyamino acid, hydrophobic vinyl-based polymer, and combinations thereof, but is limited thereto no.

In another embodiment of the present invention, the nanoparticles can control the cancer microenvironment to enhance the penetration of T cells or anticancer agents into the tumor, but is not limited thereto.

In another embodiment of the present invention, the control of the cancer microenvironment includes degradation of the extracellular matrix of the cancer microenvironment; inhibition of the production of extracellular matrix in the cancer microenvironment; inhibition of expression of tumor-associated fibroblasts; Or it may be a combination thereof, but is not limited thereto.

In another embodiment of the present invention, the nanoparticles may suppress the exosome secretion of cancer cells by releasing the endothelin receptor antagonist encapsulated therein from the cancer tissue, but is not limited thereto.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the nanoparticles as an active ingredient.

Furthermore, the present invention provides a method for preventing or treating cancer comprising administering the nanoparticles to an individual in need thereof.

In addition, the present invention provides the use of the nanoparticles for preventing, improving or treating cancer.

Furthermore, the present invention provides the use of said nanoparticles for the manufacture of an agent for the prevention, amelioration or treatment of cancer.

In one embodiment of the present invention, the cancer is lung cancer, gastric cancer, glioma, liver cancer, melanoma, kidney cancer, urothelial cancer, head and neck cancer, Merkel-cell carcinoma, prostate cancer, blood cancer, breast cancer, It may be selected from the group consisting of colorectal cancer, colon cancer, rectal cancer, pancreatic cancer, brain cancer, ovarian cancer, bladder cancer, bronchial cancer, skin cancer, cervical cancer, endometrial cancer, esophageal cancer, thyroid cancer, bone cancer, and combinations thereof, but is limited thereto no.

In addition, the present invention provides a pharmaceutical composition for enhancing the anticancer effect of an anticancer agent comprising the nanoparticles as an active ingredient.

Furthermore, the present invention provides a method for enhancing the anticancer effect of an anticancer agent comprising administering the nanoparticles to an individual in need thereof.

In addition, the present invention provides a use for enhancing the anticancer effect of the anticancer agent of the nanoparticles.

Furthermore, the present invention provides the use of the nanoparticles for the preparation of a preparation for enhancing the anticancer effect of an anticancer agent.

In one embodiment of the present invention, the anticancer agent may be selected from the group consisting of antibodies, proteins, peptides, polypeptides and compounds, but is not limited thereto.

In another embodiment of the present invention, the anticancer agent may be an immune checkpoint inhibitor, but is not limited thereto.

In another embodiment of the present invention, the immune checkpoint inhibitor may be an antibody that specifically binds to one or more immune checkpoint proteins selected from the group consisting of PD-1, PD-L1 and CTLA-4, but is not limited thereto. it is not

In addition, the present invention provides a pharmaceutical composition for inhibiting cancer metastasis comprising the nanoparticles as an active ingredient.

Furthermore, the present invention provides a method for inhibiting cancer metastasis comprising administering the nanoparticles to an individual in need thereof.

In addition, the present invention provides a use for inhibiting cancer metastasis of the nanoparticles.

Furthermore, the present invention provides the use of said nanoparticles for the preparation of an agent for inhibiting cancer metastasis.

The nanoparticles of the present invention accumulate in cancer tissues and release endothelin receptor antagonists, and simultaneously act on tumor-related fibroblasts, endothelial cells, and cancer cells that play a major role in the tumor microenvironment, thereby inhibiting the fibrosis process and reducing the extracellular matrix. By controlling the microenvironment within the tumor through inhibition of production, the response rate and therapeutic effect of anticancer drugs such as immune checkpoint inhibitors can be significantly improved.

1a to 1c are diagrams showing the physicochemical properties ( FIGS. 1a and 1b ) and cytotoxicity ( FIG. 1c ) analysis results of the nanoparticles of the present invention.
Figures 2a to 2f are roads showing the anticancer treatment efficacy of the nanoparticles of the present invention, Figure 2a shows the experimental schedule, Figure 2b shows the tumor volume change in each substance administration group over time after tumor inoculation, Figure 2c shows the change in body weight of each substance administration group over time after tumor inoculation, FIG. 2d shows the change in tumor volume for each animal in a cancer animal model in which each substance is administered intravenously, and FIG. 2e shows each Shows the tumor weight distribution in the substance-administered group, and FIG. 2f shows the histologic evaluation results according to the administration of each substance obtained by performing H&E staining on the main organs and cancer tissues of the cancer animal model.
3A to 3E are roads showing the control ability of each substance administration group for the extracellular matrix component of the tumor microenvironment and tumor-related fibroblasts, and FIGS. 3A and 3B are collagen production confirmed by masson's trichrome staining of cancer tissues of each substance administration group 3c shows the results of confirming the degree of fibronectin formation in cancer tissues of each substance-administered group using immunofluorescence, and FIGS. 3d and 3e show the expression level of tumor-associated fibroblasts in cancer tissues. The results confirmed using the fluorescence method are shown.
4a to 4e are roads showing the ability of the nanoparticles of the present invention to control the tumor microenvironment, and FIGS. 4a and 4b show the results of confirming the hypoxic environment level of the cancer tissues of each substance administration group using immunofluorescence, FIG. 4c shows the results for the solid stress proportional to the degree of spread when the cancer tissue is cut, FIG. 4d shows the amount of TGF-β1 in the cancer tissue measured in each substance administration group, and FIG. Exosomes are isolated from blood, and the level of exosome PD-L1 is measured by ELISA.
Figures 5a to 5c are diagrams showing the analysis results of the T cell population in the cancer tissue in the body after the co-administration of the nanoparticles and the immune checkpoint inhibitor of the present invention, Figures 5a and 5b are the T cell population of the cancer tissue in each substance administration group 5c shows the results of confirming the distribution of T cells in cancer tissues of each substance-administered group using immunofluorescence.
2 to 5, each substance administration group is as follows:
DPBS: saline administration group,
MCT: endothelin receptor antagonist drug administration group,
M-NP: endothelin receptor antagonist of the present invention-encapsulated nanoparticles administered group,
Ab: anti-PD-1 antibody administration group,
MCT+Ab: endothelin receptor antagonist drug and anti-PD-1 antibody co-administered group;
M-NP+Ab: The group administered in combination with the endothelin receptor antagonist-encapsulated nanoparticles of the present invention and an anti-PD-1 antibody.
6 is a schematic diagram of the endothelin receptor antagonist-encapsulated nanoparticles that control the tumor microenvironment of the present invention and a diagram showing an immune checkpoint inhibitor-based anticancer treatment method using the same.
7A and 7B are diagrams showing the cancer metastasis inhibitory effect of the nanoparticles of the present invention, FIG. 7A shows the results of a cell migration assay, and FIG. 7B shows the results of a cell invasion assay.

The present invention provides nanoparticles comprising a hydrophilic polymer and a biocompatible amphiphilic polymer comprising a hydrophobic polymer conjugated to the hydrophilic polymer, and encapsulating an endothelin receptor antagonist therein.

As used herein, the term "endothelin receptor antagonist" refers to a drug that acts on an endothelin receptor molecule in vivo to inhibit or inhibit its function.

In the present invention, the endothelin receptor antagonist is macitentan, bosentan, ambrisentan, sulfisoxazole, BQ-123, BQ-788, zibotentan. , an endothelin receptor antagonist selected from the group consisting of sitaxentan, atrasentan, tezosentan and A192621.

In the present invention, as an endothelin receptor antagonist, an endothelin receptor antagonist that exhibits antagonistic action on both the _{endothelin A receptor (ET A} ) and the endothelin B receptor (ET _{B ) can be used.} For example, _{endothelin receptor antagonists that exhibit antagonistic action against both ET A} and ET _B include macitentan, bosentan, tezosentan, and the like.

The nanoparticles of the present invention are made of a biocompatible amphipathic polymer, and an endothelin receptor antagonist is encapsulated therein. As used herein, the term "biocompatibility" refers to a thing that combines a desired function and safety for a living body in a broad sense, and in a narrow sense it means a biological safety for a living body, that is, non-toxic and sterilizable. Therefore, in the present invention, the term "biocompatible amphipathic polymer" can be said to be a substance that exhibits a desired function in a living body, has no toxicity of the material itself, and can be sterilized.

In the present invention, the amphiphilic polymer includes a hydrophilic polymer and a hydrophobic polymer conjugated to the hydrophilic polymer. The hydrophilic polymer may be a polysaccharide polymer, for example, the polysaccharide polymer is carboxymethyl dextran, chitosan, gelatin, collagen, mannan, dextran sulfate (dextran sulfate), α-cyclodextrin (α-cyclodextrin), β-cyclodextrin, γ-cyclodextrin, hyaluronic acid, alginic acid, glycogen, amylose, beta-glucan ( β-glucan), hydroxyethyl cellulose, carboxymethyl cellulose, fucoidan, chondroitin, and combinations thereof.

In addition, the hydrophilic polymer is, for example, polyethylene glycol, polyethylene oxide, polyoxazoline, poly(N-vinylpyrrolidone), polyvinyl alcohol, polyhydroxyethyl methacrylate, polyserine, polythreonine, polytyrosine, polylysine, polyarginine, polyhistidine, polyaspartic acid, polyglutamic acid, and combinations thereof.

In addition to the hydrophilic polymers listed above, any hydrophilic polymer that is available or used in the preparation of nanoparticles may be used in the preparation of the nanoparticles of the present invention without limitation.

In the present invention, the hydrophobic polymer is, for example, polylactic-co-glycolic acid (Poly(lactic-co-glycolic acid), polyphosphazene, polylactide, polycaprolactone, polyanhydride, polymalic acid, polyalkylcyanoacrylate, polyhydrooxybutylate, polycarbonate, polyorthoester, hydrophobic polyamino acid, hydrophobic vinyl-based polymer, and combinations thereof. In addition to the hydrophobic polymer, any hydrophobic polymer that is available or used in the preparation of nanoparticles may be used for the preparation of the nanoparticles of the present invention without limitation.

The nanoparticles of the present invention can control the cancer microenvironment to enhance the penetration of T cells or anticancer agents into the tumor. By enhancing the infiltration of the T cells and the anticancer agent into the tumor, it is possible to enhance the apoptosis effect of the T cells and the cancer cell by the anticancer agent.

In the present invention, the control of the cancer microenvironment includes degradation of the extracellular matrix of the cancer microenvironment; inhibition of the production of extracellular matrix in the cancer microenvironment; inhibition of expression of tumor-associated fibroblasts; or a combination thereof.

In addition, the nanoparticles of the present invention can suppress the exosome secretion of cancer cells by releasing the endothelin receptor antagonist encapsulated therein from the cancer tissue. The term, “inhibition of exosome secretion” refers to inhibition of exosome production and/or secretion in cancer. Cancer generates and secretes exosomes, and the secreted exosomes contain substances such as proteins (eg, PD-L1) necessary to suppress the anticancer immune response. By secreting such cancer exosomes, the anticancer immune response is suppressed, and the therapeutic effect of the immune checkpoint inhibitor is reduced. In addition, exosome PD-L1 is known to be the main cause of suppressing the function of T cells through the bloodstream. In one embodiment of the present invention, the level of exosomal PD-L1 present in the exosomes extracted from the blood of the tumor animal model was measured, and as a result of the measurement, the administration of nanoparticles encapsulating the endothelin receptor antagonist of the present invention It was confirmed that the secretion of cancer exosomes was inhibited by (see Example 4).

As another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the nanoparticles of the present invention as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for enhancing the anticancer effect of an anticancer agent comprising the nanoparticles of the present invention as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for inhibiting cancer metastasis comprising the nanoparticles of the present invention as an active ingredient.

In addition, the present invention provides a method for preventing or treating cancer, a method for enhancing the anticancer effect of an anticancer agent, and a method for inhibiting cancer metastasis, comprising administering the pharmaceutical composition to an individual in need thereof.

As used herein, the term "cancer" refers to a problem in the control function of normal division, differentiation, and death of cells, resulting in abnormally excessive proliferation, infiltrating surrounding tissues and organs to form cancerous tissue, and destroying existing structures It means a state of being or transforming. The cancer may be divided into a primary cancer existing at the site of occurrence and a metastatic cancer that has spread from the site of occurrence to other organs in the body.

As used herein, the term "subject" refers to a subject in need of treatment for a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat, horse. , and mammals such as cattle.

In the present invention, "administration" means providing a given composition of the present invention to a subject by any suitable method.

In the present invention, "prevention" means any action that inhibits or delays the onset of a desired disease, and "treatment" means that the desired disease and metabolic abnormalities are improved or It means all actions that are advantageously changed, and "improvement" means all actions that reduce the parameters related to the desired disease, for example, the degree of symptoms by administration of the composition according to the present invention.

As used herein, the term “metastasis” refers to a condition in which a malignant tumor has spread to other tissues away from the diseased organ. As a malignant tumor that starts in one organ progresses, it spreads from the primary site, the organ, to other tissues. Metastasis can be said to be a phenomenon accompanying the progression of malignant tumors, and metastasis can occur while malignant tumor cells proliferate and acquire new genetic traits as the cancer progresses. When tumor cells that have acquired new genetic traits invade blood vessels and lymph glands, circulate through blood and lymph, and settle and proliferate in other tissues, metastasis can occur. Depending on the tissue in which metastasis occurs, various cancer diseases such as liver cancer, kidney cancer, lung cancer, stomach cancer, colorectal cancer, rectal cancer, and pancreatic cancer may be induced. The composition according to the present invention can prevent and treat the spread of cancer by inhibiting metastasis.

As used herein, the term "inhibition" refers to any action of inhibiting the cancer metastasis by administration of the composition according to the present invention.

In the present invention, the cancer is lung cancer, stomach cancer, glioma, liver cancer, melanoma, kidney cancer, urothelial cancer, head and neck cancer, Merkel-cell carcinoma, prostate cancer, blood cancer, breast cancer, colorectal cancer, colon cancer , rectal cancer, pancreatic cancer, brain cancer, ovarian cancer, bladder cancer, bronchial cancer, skin cancer, cervical cancer, endometrial cancer, esophageal cancer, thyroid cancer, bone cancer, and combinations thereof. Since the nanoparticles of the present invention control the cancer microenvironment to enhance the infiltration of T cells into the tumor, and thus obtain an anticancer effect by T cells, the pharmaceutical composition of the present invention is suitable for all types of cancer regardless of the type of cancer. may be effective against cancer.

In addition, when the pharmaceutical composition of the present invention is administered in combination with other anticancer agents, the nanoparticles contained in the pharmaceutical composition of the present invention control the cancer microenvironment to enhance penetration of the co-administered anticancer agent into cancer tissues, so that the anticancer agent It is possible to enhance the cancer treatment effect of an anticancer agent administered in combination without limitation in the type of the drug. Therefore, the range of the anticancer agent that can be administered in combination with the pharmaceutical composition of the present invention may be included without limitation as long as it acts on cancer cells or cancer tissues, such as antibodies, proteins, peptides, polypeptides and compounds, and exhibits anticancer effects.

For example, the anticancer agent includes mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine. (carmustine: BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin: actinomycin D), doxorubicin (adriamycin), daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin, C breomycin C Bleomycin, vincristine, vinblastine, paclitaxel, docetaxel, etoposide, teniposide, topotecan and iridotecan ( iridotecan), etc.

In the present invention, the anticancer agent may be an immune checkpoint inhibitor. As used herein, the term "immune checkpoint inhibitor" refers to a substance that inhibits, interferes with, or modulates, in whole or in part, one or more immune checkpoint proteins. Immune checkpoint proteins regulate the activation or function of T cells. A number of immune checkpoint proteins are known, such as PD-1, PD-L1 and CTLA-4 (Nature Reviews Cancer 12: 252-264, 2012). These proteins are involved in co-stimulatory or inhibitory interactions of T cell responses. Immune checkpoint inhibitors include, and may be derived from, antibodies.

In the present invention, the immune checkpoint inhibitor antibody may be an antibody that specifically binds to PD-1 (Programmed cell death 1) or PD-L1 (PD-1 ligand 1). For example, the antibody that specifically binds to PD-1 may be an antibody selected from the group consisting of pembrolizumab, nivolumab, and semiplimab. Also, for example, the antibody that specifically binds to PD-L1 may be an antibody selected from the group consisting of atezolizumab, avelumab, and durvalumab.

In the present invention, the immune checkpoint inhibitor antibody may be an antibody that specifically binds to cytotoxic T-lymphocyte-associated protein 4 (CTLA4). For example, the antibody that specifically binds to CTLA4 may be Ipilimumab.

As used herein, the term “antibody” refers to a substance that specifically binds to immune checkpoint proteins such as PD-1, PD-L1, CTLA4, and exhibits immune checkpoint inhibitory activity. In the antibody-drug conjugate of the present invention, the scope of the antibody conjugated to the drug includes not only the complete antibody but also the antigen-binding site of the antibody molecule.

The pharmaceutical composition of the present invention may be administered simultaneously (simultaneous), separately (separate) or sequentially (sequential) with an anticancer agent (eg, an immune checkpoint inhibitor), and may be administered single or multiple.

The content of the active ingredient in the pharmaceutical composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50, based on the total weight of the composition. It may be a weight %, but is not limited thereto. The content ratio is a value based on the dry amount from which the solvent is removed.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, at least one selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a humectant, a film-coating material, and a controlled-release additive.

The pharmaceutical composition according to the present invention can be prepared according to a conventional method, respectively, in powders, granules, sustained-release granules, enteric granules, liquids, eye drops, elsilic, emulsions, suspensions, alcohols, troches, fragrances, and limonaade. , tablets, sustained release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusates, Warnings, lotions, pasta, sprays, inhalants, patches, sterile injection solutions, or external preparations such as aerosols can be formulated and used, and the external preparations are creams, gels, patches, sprays, ointments, warning agents , lotion, liniment, pasta, or cataplasma.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid as additives for tablets, powders, granules, capsules, pills, and troches according to the present invention Calcium monohydrogen, calcium sulfate, sodium chloride, sodium hydrogen carbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methyl excipients such as cellulose, 1928, 2208, 2906, 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch powder, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodite, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, 6000, liquid paraffin, hydrogenated soybean oil (Lubri) wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, Lubricating agents such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid may be used.

As additives for the liquid formulation according to the present invention, water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (Twinester), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

In the syrup according to the present invention, a sucrose solution, other sugars or sweeteners may be used, and if necessary, a fragrance, colorant, preservative, stabilizer, suspending agent, emulsifying agent, thickening agent, etc. may be used.

Purified water may be used in the emulsion according to the present invention, and if necessary, an emulsifier, preservative, stabilizer, fragrance, etc. may be used.

Suspending agents such as acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose, 1828, 2906, and 2910 may be used in the suspending agent according to the present invention, If necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

Injectables according to the present invention include distilled water for injection, 0.9% sodium chloride injection, ring gel injection, dextrose injection, dextrose + sodium chloride injection, PEG (PEG), lactated ring gel injection, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; Solubilizing aids such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tweens, nijeongtinamide, hexamine, and dimethylacetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, buffers such as albumin, peptone, gum; isotonic agents such as sodium chloride; sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), stabilizers such as ethylenediaminetetraacetic acid; sulphating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, acetone sodium bisulfite; analgesic agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; suspending agents such as SiMC sodium, sodium alginate, Tween 80, or aluminum monostearate.

The suppository according to the present invention includes cacao fat, lanolin, witepsol, polyethylene glycol, glycerogelatin, methyl cellulose, carboxymethyl cellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lanet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolene (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Butyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium, A, AS, B, C, D, E, I, T, Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), supostal (N, Es), Wecobi (W, R, S, M, Fs), tester triglyceride base (TG-95, MA, 57) and The same mechanism may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and type of the patient's disease; Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined. In consideration of all of the above factors, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration can be contemplated, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal It can be administered according to internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, and the like.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with several related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Experimental materials and methods

1. Particle Preparation

A nanoprecipitation method was used to prepare the particles of the present invention. Specifically, PEG-PLGA (10 mg) was dissolved in 1 ml of acetonitrile, which was added dropwise to ultrapure water (2 ml), and then acetonitrile was evaporated. After 0.2 μm filtering, centrifugation was performed to obtain nanoparticles (named as NP). In the case of endothelin receptor antagonist-encapsulated nanoparticles (named M-NP), 1 mg of endothelin receptor antagonist macitentan (MCT) is dissolved in acetonitrile (0.5 ml) in ultrapure water containing PEG-PLGA, acetonitrile cosolvent was poured into

2. Physicochemical characterization

In order to analyze the size distribution of the prepared particles, NPs and M-NPs were dispersed in PBS at a concentration of 1 mg/ml, and the size distribution of the particles was confirmed through dynamic light scattering measurement. In order to analyze the shape of the particle, it was stained with uranyl acetate and the shape of the particle was observed through a transmission electron microscope. In order to evaluate the optical properties of the prepared particles, PLGA, MCT, and M-NP were dissolved in acetonitrile and NaOH cosolvent, and absorbance was measured using a UV-Vis spectrophotometer.

In order to confirm the cytotoxicity of the prepared particles, 4T1 cells were seeded in a 96-well plate at ^{the number of 1 x 10 4} /well. Each well was treated with MCT, NP, and M-NP at a concentration of 12.5, 20, 50, and 100 μg/ml, and then cultured for 24 hours. Thereafter, cytotoxicity was evaluated by MTT assay.

3. Animal test evaluation

To measure the size of the cancer animal model, the major axis ( a ) and the minor axis (b ) were measured using a caliper, and the arm volume was calculated as follows.

Arm volume (v) = a x b ² x 0.5

Animals were weighed at the same time every day using a scale. The weight of the cancer was measured by using a scale after sacrificing the animal after the experiment was finished, as shown in FIG.

4. Tumor microenvironment analysis

In order to confirm the distribution of collagen in the cancer tissue, the cancer tissue was cut into sections, and masson's trichrome staining was performed, and the collagen region was observed using an optical microscope. Then, using the ImageJ program, collagen ⁺ area was quantified and plotted. To identify fibronectin, a major component of the extracellular matrix in cancer tissue, fibroblasts, and a hypoxic environment, each marker was used to observe the change in expression level through immunofluorescence staining. To this end, the cancer tissue was prepared in sections by frozen sections, and the samples were observed using a confocal fluorescence microscope after labeling with fibronectin, αSMA, and HIF-1α antibodies conjugated with fluorescence, respectively. Quantitative analysis was performed using the ImageJ program. To quantify TGF-β1 in cancer tissues, the cancer tissues were ^{made into a homogeneous suspension using the gentleMACS TM} Dissociator, and the total protein amount was quantified through BCA assay. Thereafter, the level of TGF-β1 present in each cancer tissue was verified using the TGF-β1 ELISA kit.

To verify exosomal PD-L1 (exosomal PD-L1), plasma was isolated from mouse blood. After centrifugation at 2,000 g for 20 minutes, centrifugation was performed again at 16,500 g for 45 minutes to remove cell residues and microvesicles. Thereafter, the exosomes were isolated using an exosome separation kit (Invitrogen). Thereafter, exosomal PD-L1 was quantified through PD-L1 ELISA assay (R&D systems).

5. Immune cell analysis in the tumor microenvironment

To analyze immune cells in the tumor microenvironment, tumor-infiltrating lymphocytes were isolated using CD45 beads after dissection of cancer tissue. Thereafter, the isolated lymphocytes were stained with CD3 and CD8, and a colony of tumor-infiltrating cytotoxic T cells was confirmed using a flow cytometer. Quantitative results were analyzed using the ^{Guava easyCyte TM program.} In addition, the cancer tissues were stained with CD3 and CD8 through immunochemical staining, and the distribution of tumor-infiltrating cytotoxic T cells was confirmed using a confocal fluorescence microscope.

6. Verification of cancer metastasis inhibitory effect

In order to verify the cancer metastasis inhibitory effect, a cell migration assay was performed. ^{To this end, 1.5 x 10 5} of 4T1 cells were aliquoted into the transwell chamber (corning), and the medium containing the samples such as 500 nM MCT and ET-1 was treated in the lower well. Then incubated for 15 hours. After incubation, the cells were fixed using a fixing solution, stained using a Giemsa staining solution, and observed using an optical microscope. In the case of cell invasion assay, the transwell chamber was coated with Matrigel (300 μg/ml), and then the same procedure as in cell migration assay was performed.

Experiment result

Example 1. Preparation of endothelin receptor antagonist-encapsulated nanoparticles (M-NP) and analysis of physicochemical properties and cytotoxicity

The size distribution and shape of the prepared nanoparticles were measured using a Zetasizer (Zetasizer Nano ZS90, Malvern Instruments, UK) and a transmission electron microscopy (TEM; Talos F200X; FEI Company, USA).

As a result, as shown in FIG. 1a , the nanoparticles exhibited monodispersity in aqueous solution before and after encapsulation of macitentan, and the diameter was confirmed to be around 50 nm ( FIG. 1a ). In addition, as a result of confirming the shape of the particles through a TEM photograph, spherical particles were formed (FIG. 1a).

In the optical evaluation using a UV-Vis spectrophotometer, it was confirmed that MCT and M-NP characteristically exhibited absorption at 265 nm (FIG. 1b). This is due to the absorption characteristics of MCTs, which means that MCTs were successfully encapsulated in the PEG-PLGA polymer. On the other hand, the PEG-PLGA polymer did not show absorption at 265 nm.

In addition, as a result of confirming cytotoxicity, as shown in FIG. 1c , the PEG nanoparticles (NP) that did not encapsulate the drug did not induce toxicity to the breast cancer cell line (4T1), and macitentan and macitentan-encapsulated nanoparticles showed a behavior inducing cytotoxicity when treated at high concentrations (FIG. 1c).

Example 2. Evaluation of anticancer treatment efficacy of M-NP in disease animal models

In order to evaluate the efficacy of immuno-cancer treatment using the nanoparticles (M-NP) of the present invention, a tumor animal model was prepared. Briefly, a breast cancer animal model was prepared by inoculating 4T1 cells, a breast cancer cell line, into the fat pad of mouse breast (2 X 10 ^{5 cells).} M-NP was intravenously injected at a dose of 1 mg/kg based on macitentan, and the control group was intravenously injected with only saline or macitentan (MCT) at a dose of 1 mg/kg. In addition, the anti-PD-1 antibody was intraperitoneally injected at a dose of 5 mg/kg once every 3 days to the group administered with the antibody combination to evaluate the therapeutic efficacy with the group to which the antibody was not administered. The specific experimental schedule is shown in FIG. 2A.

As a result of the experiment, in the case of the combined administration group administered with both M-NP and PD-1 antibody, the tumor volume was suppressed about 10 times compared to the control group administered with saline, and even when compared with the group administered with the anti-PD-1 antibody alone The tumor volume was suppressed about 5 times, indicating high cancer treatment efficacy ( FIGS. 2B , 2D and 2E ). There was no significant difference in body weight in all administration groups (Fig. 2c).

Major organs and cancer tissues were extracted and stained with H&E (hematoxylin and eosin) staining method for histopathological analysis. A wide area of cell death was observed in comparison with other administration groups (FIG. 2f).

The above experimental results show that cancer growth can be effectively inhibited when M-NP and antibody are co-administered.

Example 3. Evaluation of the effect of M-NP on tumor microenvironment components in disease animal models

In order to evaluate the tumor microenvironment control ability of the M-NP of the present invention, after sacrificing an animal model that has been evaluated for treatment efficacy in a disease animal model, masson's trichrome staining is performed on the cancer tissues of each administration group, and the collagen region is Comparative evaluation was performed. As a result of the experiment, as a result of comparing the region of collagen, which is a major component of the extracellular matrix of the tumor microenvironment, the collagen region formed in the cancer tissue of the M-NP-administered group was about 3 times greater than that of the other administration group to which M-NP was not administered. low ( FIGS. 3A and 3B ).

In addition, as a result of confirming the expression of fibronectin, a major component of the extracellular matrix, in cancer tissues using immunofluorescence, the formation of fibronectin was significantly reduced in the M-NP-administered group (FIG. 3c).

Furthermore, the expression of tumor-associated fibroblasts, a major component of the tumor microenvironment, was observed using immunofluorescence method. expressed ( FIGS. 3D and 3E ).

Example 4. Evaluation of tumor microenvironment control ability of M-NP in disease animal model

In order to evaluate the effect of the nanoparticles of the present invention on the hypoxic environment, solid stress, cytokine and exosome secretion, the tumor was excised after sacrificing the animal model in which the treatment efficacy evaluation was completed in the disease animal model.

First, in order to evaluate the change in the hypoxic environment, which is a characteristic of the tumor microenvironment, each tumor was stained by immunofluorescence and evaluated. As a result, it was confirmed that the tumor of the M-NP-administered group had a hypoxic environment at a level about 4 times lower than that of the other substance-administered group ( FIGS. 4a and 4b ).

Accordingly, in order to confirm the change in solid stress due to the reduction in fibrosis of the tumor tissue, an opening test of each tumor was performed (FIG. 4c). As a result, it was confirmed that the solid stress of the tumor in the M-NP-administered group was about 3 times or less.

In addition, the amount of TGF-β1, one of the cytokines that promotes the fibrosis process of the tumor, makes the composition of the extracellular matrix to a high density, and promotes the activity of tumor-associated fibroblasts, was measured. As a result, it was confirmed that the secretion of TGF-β1 in the tumor was suppressed in the M-NP-administered group in which the production of tumor-associated fibroblasts was suppressed ( FIG. 4d ).

Next, in order to confirm the secretion of exosomal PD-L1 (exosomal PD-L1) from the exosomes that inhibit the cancer treatment effect of the immune checkpoint inhibitor, the exosomes were extracted from the blood of the tumor animal model and confirmed by ELISA. As a result, in the M-NP-administered group, the amount of PD-L1 present in exosomes in the blood was reduced by about 2 times, which is because macitentan encapsulated in nanoparticles acts on cancer cells to suppress the secretion of exosomes. (Fig. 4e).

Example 5. Evaluation of changes in T cell population and tumor distribution according to M-NP administration

In order to evaluate the efficacy of the combined treatment of nanoparticles, the nanoparticles of the present invention and an immune checkpoint inhibitor were administered in combination in an animal model of breast cancer. As a result, it was confirmed that when the immune checkpoint inhibitor and the nanoparticles of the present invention were co-administered, ^{the population of CD8 +} T cells was increased by about 3 times compared to the saline-administered group ( FIGS. 5a and 5b ).

In addition, as a result of confirming the distribution of T cells in cancer tissues using the immunofluorescence method, it was confirmed that the T cell distribution was wider in the group administered with M-NP and the immune checkpoint inhibitor compared to the group administered with only the immune checkpoint inhibitor. (Fig. 5c).

Through the above results, nanoparticles containing macitentan, an endothelin receptor antagonist, simultaneously act on tumor-related fibroblasts, endothelial cells, and cancer cells, which play a major role in the tumor microenvironment, thereby inhibiting the fibrosis process and inhibiting the production of extracellular matrix composition. By controlling the microenvironment within the tumor through the control of the tumor microenvironment, the response rate and therapeutic effect of immune checkpoint inhibitors are enhanced, and accordingly, it is expected to be applicable to the development of various combination treatment technologies.

Example 6. Cancer metastasis inhibitory effect

In order to investigate the cancer metastasis inhibitory effect of M-NP, an in vitro migration and invasion assay was performed. As a result, as shown in FIG. 7a , M-NP inhibited ET-1-dependent migration of 4T1 cells (M-NP: about 2 times compared to non-treated). In a similar trend as in the cell migration assay, M-NP effectively inhibited the invasion of 4T1 cells in the invasion assay ( FIG. 7b ). These results show the cancer metastasis inhibitory effect of M-NP.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (16)

Nanoparticles comprising a hydrophilic polymer and a biocompatible amphiphilic polymer comprising a hydrophobic polymer conjugated to the hydrophilic polymer, and encapsulating an endothelin receptor antagonist therein. According to claim 1,
The endothelin receptor antagonist is an endothelin A receptor (ET _A ) and endothelin B receptor (ET _B ) Characterized in that it exhibits antagonistic action against both, nanoparticles. According to claim 1,
The endothelin receptor antagonist is macitentan, bosentan, ambrisentan, sulfisoxazole, BQ-123, BQ-788, zibotentan, sitaxentan ), atrasentan (atrasentan), tezosentan (tezosentan) and A192621, characterized in that selected from the group consisting of, nanoparticles. According to claim 1,
The hydrophilic polymer is carboxymethyl dextran, chitosan, gelatin, collagen, mannan, dextran sulfate, α-cyclodextrin (α-cyclodextrin) , β-cyclodextrin, γ-cyclodextrin, hyaluronic acid, alginic acid, glycogen, amylose, beta-glucan, hydroxyethyl cellulose, Nanoparticles, characterized in that the polysaccharide polymer is selected from the group consisting of carboxymethyl cellulose, fucoidan, chondroitin, and combinations thereof. According to claim 1,
The hydrophilic polymer is polyethylene glycol, polyethylene oxide, polyoxazoline, poly(N-vinylpyrrolidone), polyvinyl alcohol, polyhydroxyethyl methacrylate, polyserine, polythreonine, polytyrosine, polylysine, poly Nanoparticles, characterized in that selected from the group consisting of arginine, polyhistidine, polyaspartic acid, polyglutamic acid, and combinations thereof. According to claim 1,
The hydrophobic polymer is a polylactic acid-glycolic acid copolymer (Poly (lactic-co-glycolic acid), polyphosphazene, polylactide, polycaprolactone, polyanhydride, polymalic acid, polyalkylcyanoacrylate , Polyhydrooxybutyrate, polycarbonate, polyorthoester, hydrophobic polyamino acid, hydrophobic vinyl-based polymer, characterized in that selected from the group consisting of a combination thereof, nanoparticles. According to claim 1,
The nanoparticles control the cancer microenvironment, characterized in that it enhances the penetration of T cells or anticancer agents into the tumor, nanoparticles. 8. The method of claim 7,
The control of the cancer microenvironment may include degradation of the extracellular matrix of the cancer microenvironment; inhibition of the production of extracellular matrix in the cancer microenvironment; inhibition of expression of tumor-associated fibroblasts; Or, characterized in that a combination thereof, nanoparticles. According to claim 1,
The nanoparticles are characterized in that by releasing the endothelin receptor antagonist encapsulated therein from the cancer tissue to suppress the exosome secretion of cancer cells, nanoparticles. A pharmaceutical composition for the prevention or treatment of cancer comprising the nanoparticles of any one of claims 1 to 9 as an active ingredient. 11. The method of claim 10,
The cancer is lung cancer, stomach cancer, glioma, liver cancer, melanoma, kidney cancer, urothelial cancer, head and neck cancer, Merkel-cell carcinoma, prostate cancer, blood cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, pancreatic cancer, A pharmaceutical composition, characterized in that it is selected from the group consisting of brain cancer, ovarian cancer, bladder cancer, bronchial cancer, skin cancer, cervical cancer, endometrial cancer, esophageal cancer, thyroid cancer, bone cancer, and combinations thereof. A pharmaceutical composition for enhancing the anticancer effect of an anticancer agent comprising the nanoparticles of any one of claims 1 to 9 as an active ingredient. 13. The method of claim 12,
The anticancer agent is characterized in that selected from the group consisting of antibodies, proteins, peptides, polypeptides and compounds, a pharmaceutical composition. 13. The method of claim 12,
The anticancer agent is an immune checkpoint inhibitor (immune checkpoint inhibitor), characterized in that the pharmaceutical composition. 15. The method of claim 14,
The immune checkpoint inhibitor is an antibody that specifically binds to one or more immune checkpoint proteins selected from the group consisting of PD-1, PD-L1 and CTLA-4, a pharmaceutical composition. A pharmaceutical composition for inhibiting cancer metastasis, comprising the nanoparticles of any one of claims 1 to 9 as an active ingredient.

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