Composition containing narciclasine or a pharmaceutically or sitologically acceptable salt thereof for prevention, alleviation, or treatment of cancer

Abstract

The present invention relates to a composition containing narciclasine or a pharmaceutically or sitologically acceptable salt thereof for the prevention, alleviation, or treatment of cancer having mutant EGFR, or a composition for the prevention, alleviation, or treatment of cancer resistant to tyrosine kinase inhibitors (TKIs), wherein the composition can significantly inhibit or suppress the growth of cancer cells in cancers that are resistant to tyrosine kinase inhibitors or have mutant EGFR, by inhibiting or reducing the activity or expression of EGFR by promoting EGFR degradation through a proteasomal degradation pathway or lysosomal degradation pathway, and thus can be usefully employed to prevent or treat cancer.

Description

Composition for preventing, improving or treating cancer comprising narcyclasin or a pharmaceutically or food-wise acceptable salt thereof

[0001] The present invention relates to a composition for preventing, improving or treating cancer having an EGFR mutant type, comprising narcyclacin or a pharmaceutically or food-wise acceptable salt thereof; or a composition for preventing, improving or treating cancer resistant to a tyrosine kinase inhibitor (TKI).

[0002] EGFR (epidermal growth factor receptor) is a member of the tyrosine kinase receptor family, also known as HER or ErbB family, and is a key gene that regulates cell growth, cell survival, and migration. Overexpression or mutation of EGFR increases tyrosine kinase activity, which is known to be related to the progression of various solid cancers such as breast cancer and lung cancer. In particular, EGFR mutations are found in non-squamous cell carcinoma among non-small cell lung cancer (NCSCLC), especially adenocarcinoma. Non-squamous cell carcinoma caused by EGFR mutations is found in about 50% of Asians and about 20% of Westerners. Therefore, EGFR mutations are most commonly found in young people, women, Asians, and non-smokers with adenocarcinoma.

[0003] EGFR mutations occur in the tyrosine kinase domain of EGFR, increasing the activity of EGFR kinase, thereby continuously activating cell signaling pathways and allowing cell differentiation and growth to continue. The most common EGFR mutations are the deletion of exon 19 and the L858R mutation, which is a point mutation. These two mutations account for approximately 90% of all EGFR mutations. They are also called sensitizing EGFR mutations because they respond well to first- and second-generation EGFR tyrosine kinase inhibitor (TKI) treatment (Erminia Massarelli et al., Lung Cancer (review) 2013 Jun;80(3):235-41). However, lung cancer cells with the T790M mutation, a mutation at codon 790, are resistant to existing TKIs (gefitinib), making treatment with existing anticancer drugs difficult.

[0004] Accordingly, the inventors of the present invention completed the present invention by discovering that narcissiclasine, a single compound derived from the root extract of the common ginseng, has a strong anticancer effect on cancer with an EGFR mutant type.

[0005] The present invention aims to solve the above-mentioned problems and other problems related thereto.

[0006] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating cancer having an epidermal growth factor receptor (EGFR) mutant, comprising narcissine or a pharmaceutically acceptable salt thereof.

[0007] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer resistant to a tyrosine kinase inhibitor (TKI), comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0008] Another object of the present invention is to provide a food composition for preventing or improving cancer having an EGFR mutant type, comprising narcyclacin or a food-wise acceptable salt thereof.

[0009] Another object of the present invention is to provide a food composition for preventing or improving cancer resistant to a tyrosine kinase inhibitor (TKI) comprising narcyclacin or a food-based acceptable salt thereof.

[0010] Another object of the present invention is to provide an anticancer adjuvant comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0011] Another object of the present invention is to provide a use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer having an EGFR mutant type.

[0012] Another object of the present invention is to provide a use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer resistant to tyrosine kinase inhibitors.

[0013] Another object of the present invention is to provide a use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer having an EGFR mutant type.

[0014] Another object of the present invention is to provide a use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer resistant to tyrosine kinase inhibitors.

[0015] Another object of the present invention is to provide a method for preventing or treating cancer having an EGFR mutant type, comprising administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0016] Another object of the present invention is to provide a method for preventing or treating cancer resistant to a tyrosine kinase inhibitor, comprising administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0017]

[0018] The technical problem to be achieved according to the technical idea of ​​the invention disclosed in this specification is not limited to the problem to solve the above-mentioned problem, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0019] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in this application can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not limited by the specific descriptions described below.

[0020] The terms used in the present invention are general terms that are widely used as much as possible while taking into account the functions of the present invention, but these may change depending on the intention of a technician working in the relevant technical field or the emergence of new technologies.

[0021] Additionally, in certain cases, terms may be arbitrarily selected, and in such cases, their meanings will be described in detail in the description of the relevant embodiments. Therefore, the terms used in the present invention should be defined based on their meaning and the overall content of the present invention, rather than simply their names.

[0022] When the present invention says that a component or a step "includes", this does not mean that other components or other steps are excluded, but rather that other components or other steps may be further included, unless specifically stated otherwise.

[0023]

[0024] As one aspect for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating cancer having an epidermal growth factor receptor (EGFR) mutant, comprising narcissine or a pharmaceutically acceptable salt thereof.

[0025] In the present invention, the "narcissac" may be a compound represented by the following chemical formula 1, and may be isolated from the root extract of Sasa japonica. In addition, the narcissac of the present invention may be chemically synthesized using a method known in the art or may use a commercially available material.

[0026] [Chemical Formula 1]

[0027]

[0028] In the present invention, the pharmaceutically acceptable salt of narcyclacin is useful as an acid addition salt formed by a pharmaceutically acceptable free acid or a metal salt formed by a base. As an example, the free acid may be an inorganic acid or an organic acid, and the inorganic acid may be hydrochloric acid, sulfuric acid, hydrobromic acid, sulfurous acid, or phosphoric acid, and the organic acid may be citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, or methanesulfonic acid. In addition, the metal salt may be an alkali metal salt or an alkaline earth metal salt, sodium, potassium, or calcium salt. However, the present invention is not necessarily limited thereto.

[0029] In the present invention, the "cancer" may be a solid cancer having an EGFR (epidermal growth factor receptor) mutation, and the solid cancer may be selected from the group consisting of lung cancer, breast cancer, colon cancer, colon cancer, pancreatic cancer, kidney cancer, head and neck cancer, ovarian cancer, cervical cancer, prostate cancer, skin cancer, liver cancer, thyroid cancer, gallbladder cancer, bile duct cancer, esophageal cancer, bladder cancer, and oral cancer, and preferably, the solid cancer may be lung cancer, but is not limited thereto.

[0030] Additionally, the lung cancer may be selected from the group consisting of non-small cell lung cancer, adenocarcinoma, squamous cell carcinoma, large cell carcinoma, and small cell lung cancer, but is not limited thereto.

[0031] In the present invention, the "EGFR mutation" may include one or more mutations selected from the group consisting of deletion of EGFR exon 19, L858R mutation, T790M mutation, and C797S mutation, and preferably, may include one or more mutations selected from the group consisting of L858R mutation and T790M mutation.

[0032] The above L858R mutation refers to a mutation in which the 858th amino acid of the EGFR protein is substituted from L (leucine) to R (arginine), and the above T790M mutation refers to a mutation in which the 790th amino acid of the EGFR protein is substituted from T (threonine) to M (methionine). The sequence of the EGFR protein can be referenced from information such as UniProt (P00533).

[0033] In the present invention, the composition may inhibit or reduce the activity or expression of EGFR, and may promote the degradation of EGFR through the proteasomal degradation pathway or the lysosomal degradation pathway.

[0034] The present invention has confirmed that a composition containing narcyclasin has a remarkable effect in preventing or treating cancer, particularly lung cancer, by promoting the degradation of EGFR through the proteasomal degradation pathway or the lysosomal degradation pathway in cancer having L858R mutation, T790M mutation, or L858R / T790M double mutation of EGFR, thereby reducing the activity or expression of EGFR.

[0035] In the present invention, the term “prevention” refers to any act of suppressing or delaying symptoms of cancer by administering the composition of the present invention.

[0036] In the present invention, the term “treatment” means any act in which the symptoms of cancer are improved or beneficially changed by administering the composition of the present invention.

[0037] As another aspect for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating cancer resistant to a tyrosine kinase inhibitor (TKI) comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0038] The above “narcyclasin”, “pharmaceutically acceptable salt”, “cancer”, “prevention”, “treatment” and “composition” are as described above.

[0039] In the present invention, the "tyrosine kinase inhibitor" refers to a substance that inhibits tyrosine kinase, which is an enzyme responsible for the activation of various proteins through a signal transduction cascade. Proteins regulated by the tyrosine kinase are activated by adding a phosphate group to the protein, and the tyrosine kinase inhibitor acts by inhibiting this phosphorylation step.

[0040] In the present invention, the tyrosine kinase inhibitor is an anticancer agent, and may be, but is not limited to, Gefitinib, Erlotinib, or Afatinib.

[0041] In the present invention, the term “resistance” or “tolerance” means not exhibiting a significant cellular or biological response to a specific drug, and specifically means that the rate of death or cell death of cancer cells does not appear or decreases in response to treatment using the drug.

[0042] The composition containing narcyclacin of the present invention significantly inhibited or suppressed the growth of cancer cells in cancer cells having EGFR mutation type, cancer cells resistant to tyrosine kinase inhibitors (TKIs), particularly gefitinib, and particularly lung cancer, and it was confirmed that it had a stronger anticancer effect even at a lower concentration than the therapeutic concentration for cancer cells having EFGF wild type.

[0043] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" means that the composition exhibits non-toxic properties to cells or humans exposed to the composition. Pharmaceutically acceptable carriers may further comprise, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.

[0044] Additionally, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose, and glycols. They may also contain stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.

[0045] The pharmaceutical composition of the present invention can be administered to mammals, including humans, by any method. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, central, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal administration.

[0046] The pharmaceutical composition of the present invention may be formulated as a preparation for oral or parenteral administration, depending on the route of administration as described above. When formulated, it may be prepared using one or more buffers (e.g., saline or PBS (phosphate buffered saline)), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrating agents, or surfactants, diluents, or excipients.

[0047] Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions, capsules, etc., and these solid preparations can be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient, for example, starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl-cellulose, or gelatin. For example, tablets or sugar-coated tablets can be obtained by mixing an active ingredient with a solid excipient, grinding the mixture, adding a suitable auxiliary agent, and then processing the mixture into a granule mixture.

[0048] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water or liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, or preservatives may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as disintegrants, and anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, and preservatives may be additionally included.

[0049] When administered parenterally, the pharmaceutical composition of the present invention may be formulated in the form of injections, transdermal administration agents, and nasal inhalants together with a suitable parenteral carrier according to methods known in the art. The injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Suitable carriers for injections include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, suitable carriers include Hanks' solution, Ringer's solution, PBS containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. To protect the injections from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal may be additionally included. Additionally, the above injections may in most cases additionally contain isotonic agents such as sugar or sodium chloride.

[0050] Transdermal administration includes ointments, creams, lotions, gels, topical solutions, pastes, liniments, and aerosols. Here, "transdermal administration" means topically administering a pharmaceutical composition to the skin, thereby delivering an effective amount of the active ingredient contained in the pharmaceutical composition into the skin.

[0051] For inhalation administration, the compositions used according to the present invention may conveniently be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or another suitable gas. For pressurized aerosols, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in the well-known prescription book of pharmaceutical chemistry (Remington's Pharmaceutical Science, 15th Edition, 1975 Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour).

[0052] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects. The effective dosage level may be determined based on factors including the patient's health condition, type and severity of the disease, drug activity and sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, combination or concurrent drug use, and other factors well known in the medical field. The dosage and frequency of administration do not limit the scope of the present invention in any way.

[0053] The pharmaceutical composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, or biological response modifiers.

[0054]

[0055] As another aspect for achieving the above object, the present invention provides a food composition for preventing or improving cancer having an EGFR mutant type, comprising narcyclacin or a food-wise acceptable salt thereof.

[0056] The above “narcyclasin”, “EGFR mutation”, “cancer”, “prevention” and “composition” are as described above.

[0057] As another aspect for achieving the above object, the present invention provides a food composition for preventing or improving cancer resistant to a tyrosine kinase inhibitor (TKI) comprising narcyclacin or a food-wise acceptable salt thereof.

[0058] The above “narcyclasin”, “tyrosine kinase inhibitor”, “resistance”, “cancer”, “prevention” and “composition” are as described above.

[0059] In the present invention, the term “improvement” means that the composition of the present invention exhibits an effect of alleviating the symptoms of cancer.

[0060] In the present invention, the food-acceptable salt of narcislacin is useful as an acid addition salt formed by a food-acceptable free acid or a metal salt formed by a base. As an example, the free acid may be an inorganic acid or an organic acid, and the inorganic acid may be hydrochloric acid, sulfuric acid, hydrobromic acid, sulfurous acid, or phosphoric acid, and the organic acid may be citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, or methanesulfonic acid. In addition, the metal salt may be an alkali metal salt or an alkaline earth metal salt, or a sodium, potassium, or calcium salt. However, the present invention is not necessarily limited thereto.

[0061] The food composition of the present invention may include all foods in the conventional sense, and may be used interchangeably with terms known in the art, such as functional food and health functional food.

[0062] The term "functional food" of the present invention means a food manufactured and processed using raw materials or ingredients having functionality useful to the human body according to Act No. 6727 on Health Functional Foods, and "functionality" means consumption for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological action.

[0063] The term "health functional food" of the present invention refers to a food manufactured or processed by using a specific ingredient as a raw material or extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplementation, and refers to a food designed and processed so that the above-mentioned ingredient can sufficiently exert a bioregulatory function on the body, such as biodefense, regulation of biological rhythm, prevention and recovery from disease, etc., and the above-mentioned health food composition can perform functions related to disease prevention and disease recovery.

[0064] There is no limitation on the types of foods in which the composition of the present invention can be used. In addition, the composition of the present invention can be prepared by mixing other appropriate auxiliary ingredients that can be included in foods and known additives according to the selection of a person skilled in the art. Examples of foods to which the composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and the extract according to the present invention and its fractions can be prepared by adding them to juice, tea, jelly, and juice, etc. made with the extract and its fractions as main ingredients.

[0065] In addition, foods applicable to the present invention may include all foods, such as special nutritional foods (e.g., formula milk, infant food, etc.), processed meat products, fish products, tofu, starch jelly, noodles (e.g., ramen, noodles, etc.), health supplements, seasoned foods (e.g., soy sauce, soybean paste, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g., snacks), processed dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (e.g., various kimchi, pickled vegetables, etc.), beverages (e.g., fruit and vegetable beverages, soy milk, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.).

[0066] When the health functional food composition of the present invention is used in the form of a beverage, it may contain various sweeteners, flavoring agents, or natural carbohydrates as additional ingredients, just like conventional beverages. In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks.

[0067] As another aspect for achieving the above object, the present invention provides an anticancer adjuvant comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0068] The above “narcyclacin”, “pharmaceutically acceptable salt” and “cancer” are as described above.

[0069] The above anticancer adjuvant may be administered in combination with an anticancer drug.

[0070] The above-mentioned anticancer adjuvant refers to any form of anticancer agent that enhances the anticancer effect of an anticancer agent or suppresses or ameliorates the side effects of the agent. The anticancer adjuvant of the present invention can be administered in combination with various types of anticancer agents or anticancer adjuvants. When administered in combination, the anticancer agent can exhibit an equivalent level of anticancer therapeutic effect even at a lower dosage than conventional anticancer agents, thereby enabling safer anticancer treatment.

[0071] The above-mentioned anticancer adjuvant may be administered via any conventional route as long as it can reach the target tissue. The anticancer adjuvant of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intrapulmonary, or rectally, depending on the intended purpose, but is not limited thereto. Furthermore, the anticancer adjuvant may be administered via any device capable of transporting the active substance to target cells.

[0072] The anticancer adjuvant of the present invention can be preferably formulated as an anticancer adjuvant by additionally including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration. Carriers, excipients or diluents that can be included in the anticancer treatment adjuvant of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0073] The anticancer adjuvant of the present invention may be a formulation for parenteral administration, and the description of the formulation is replaced with the description of the formulation of the pharmaceutical composition.

[0074] In addition, any anticancer adjuvant disclosed in the present technical field can be applied to the present invention without significant limitations.

[0075]

[0076] As one aspect for achieving the above object, the present invention provides the use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer having an EGFR mutant type.

[0077] The above “EGFR mutation”, “cancer”, “prevention”, “treatment”, “narcyclasin” and “composition” are as described above.

[0078] As one aspect for achieving the above object, the present invention provides the use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer resistant to a tyrosine kinase inhibitor.

[0079] The above "tyrosine kinase inhibitor", "resistance", "cancer", "prevention", "treatment", "narcyclasin" and "composition" are as described above.

[0080] As one aspect for achieving the above object, the present invention provides the use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating cancer having an EGFR mutant type.

[0081] The above “EGFR mutation”, “cancer”, “prevention”, “treatment”, “narcyclasin” and “composition” are as described above.

[0082] As one aspect for achieving the above object, the present invention provides the use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating cancer resistant to a tyrosine kinase inhibitor.

[0083] The above "tyrosine kinase inhibitor", "resistance", "cancer", "prevention", "treatment", "narcyclasin" and "composition" are as described above.

[0084] As one aspect for achieving the above object, the present invention provides a method for preventing or treating cancer having an EGFR mutant type, comprising a step of administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0085] The above “narcyclasin”, “EGFR mutation”, “cancer”, “prevention”, “treatment” and “composition” are as described above.

[0086] As one aspect for achieving the above object, the present invention provides a method for preventing or treating cancer resistant to a tyrosine kinase inhibitor, comprising the step of administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

[0087] The above “narcyclasin”, “tyrosine kinase inhibitor”, “resistance”, “cancer”, “prevention”, “treatment” and “composition” are as described above.

[0088] The composition of the present invention can significantly inhibit or suppress the growth of cancer cells by inhibiting or reducing the activity or expression of EGFR by promoting the degradation of EGFR through the proteasomal degradation pathway or the lysosomal degradation pathway in cancer resistant to tyrosine kinase inhibitors (TKIs) or cancer with an EGFR mutation, and thus can be usefully used for the prevention or treatment of cancer.

[0089] Figure 1 shows the results of measuring the cell growth curves using a Live Cell Image device after treating A549 cell line (EGFR Wild-Type, Figure 1a), H1299 cell line (EGFR Wild-Type, Figure 1b), and H1975 cell line (EGFR mutant T790M / L858R, Figure 1c) with various concentrations (25 nM, 50 nM, 100 nM, 500 nM, 1 μM, 5 μM, and 10 μM) of narcyclasin (NCS).

[0090] Figure 2 shows the results of a simulation model predicting the binding of narcyclacin (NCS) and EGFR wild-type or mutant type (T790M / L858R).

[0091] Figure 3 shows the results of measuring kinase activity by measuring the amount of ATP consumed after treating EGFR wild-type kinase (Figure 3a) or EGFR mutant (T790M / L858R) kinase (Figure 3b) with 50 nM gefitinib (Gef) or 50 nM narcyclasin (NCS).

[0092] Figure 4 shows the results of examining the degree of cell survival after treating EGFR mutant PC-9 cell lines and gefitinib-resistant PC-9 (PC-9-GR) cell lines with narcyclasin (NCS).

[0093] Figure 5a shows the results of Western blot analysis of the expression levels of phosphorylated EGFR (p-EGFR) and total EGFR after 4, 8, and 24 hours of treatment with 50 nM narcyclacin (NCS) in H1975 cells.

[0094] Figure 5b shows the results of immunofluorescence staining analysis to confirm the expression sites of LAMP1 (green) and EGFR (red), which are markers of lysosomes that decompose intracellular substances, in H1975 cells treated with narcyclacin (NCS).

[0095] Figure 5c shows the results of comparing the expression levels of EGFR according to the presence or absence of narcyclasin (NCS) treatment after treatment with cycloheximide (CHX), a protein synthesis inhibitor.

[0096] Figure 5d shows the results of comparing the expression levels of EGFR according to the presence or absence of narcyclacin (NCS) treatment after treatment with MG132, a proteasome inhibitor, and Bafilomycin A1 (Baf A1), a lysosomal inhibitor.

[0097] Figure 6 shows the results of comparing the formation of multivulva in a C. elegans cancer model with the EGFR mutant (L858R / T790M) and wild-type C. elegans (A), confirming whether multivulva was formed after treatment with narcyclasin (NCS) (B), and comparing the relative degree of multivulva formation after treatment with gefitinib or narcyclasin (NCS) (C).

[0098] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to more specifically illustrate the present invention, and the scope of the present invention is not limited to these examples.

[0099]

[0100] Example 1. Growth inhibition effect of cancer cells by narcissiclasine

[0101] An experiment was conducted to determine whether narcissine (NCS) has an effect in inhibiting the growth of cancer cells.

[0102] A549 cell line (EGFR Wild-Type), H1299 cell line (EGFR Wild-Type), and H1975 cell line (EGFR mutant T790M / L858R) were seeded in 96-well plates at 4 x 103 / well and treated with narcyclacin (NCS) at various concentrations (25 nM, 50 nM, 100 nM, 500 nM, 1 μM, 5 μM, and 10 μM). Afterwards, the narcyclacin (NCS)-treated cells were placed in a Live Cell Image device and cultured, and the degree of cell growth was measured every 3 hours for 48 hours to draw a cell growth curve, and the survival rate for each concentration was calculated to derive the IC50 value.

[0103] As a result, as shown in Fig. 1, it was confirmed that narcyclacin (NCS) had a significant effect of inhibiting or suppressing cell growth in a concentration-dependent manner in all three types of lung cancer cell lines with EGFR wild-type or EGFR mutant (T790M / L858R). In particular, the IC50 values ​​of narcyclacin (NCS), which inhibited cell growth by 50% in the A549 cell line and H1299 cell line with EGFR wild-type, were confirmed to be 0.11 μM (Fig. 1a) and 0.08 μM (Fig. 1b), respectively, whereas in the H1975 cell line with EGFR mutant (T790M / L858R), the IC50 value was confirmed to be 0.022 μM (Fig. 1c). Thus, it was confirmed that narcyclacin (NCS) has a significant cell growth inhibition effect even at lower concentrations on cancer cell lines with EGFR mutations (T790M / L858R).

[0104] From the above results, it was confirmed that the narcyclasin (NCS) of the present invention has an effect of significantly inhibiting or suppressing the growth of cancer cells with EGFR mutant type (T790M / L858R).

[0105]

[0106] Example 2. Analysis of the three-dimensional binding structure of narcyclasin (NCS) and EGFR.

[0107] The binding structures of narcyclasin (NCS) and EGFR wild-type or mutant (T790M / L858R) were predicted using a binding simulation model using structural biology methods.

[0108] As a result, as shown in Fig. 2, it was confirmed that the binding structure of narcyclasin (NCS) and the EGFR mutant (T790M / L858R) contained more residues involved in hydrophobic interactions than the binding structure with the EGFR wild type.

[0109] From the above results, it was confirmed that the narcyclasin (NCS) of the present invention can form a stronger binding structure with the EGFR mutant type (T790M / L858R) than with the EGFR wild type (Wild-Type).

[0110]

[0111] Example 3. In vitro kinase activity reduction effect by narcyclasin (NCS)

[0112] An in vitro Kinase Assay was performed to determine whether narcyclasin (NCS) has an inhibitory or suppressive effect on EGFR wild-type and EGFR mutant (T790M / L858R).

[0113] Purified EGFR wild-type kinase and EGFR mutant (T790M / L858R) kinase were each added to an E-tube with reaction buffer, substrate, and ATP, and kinase activity was confirmed by measuring the amount of ATP consumed after incubation at 30°C after treatment with 50 nM gefitinib or 50 nM narcyclasin (NCS).

[0114] As a result, as shown in Fig. 3a, in the EGFR wild-type (WT) kinase reaction, the relative kinase activity was significantly reduced from about 8 to about 3 by treatment with 50 nM gefitinib, which was the positive control, but no significant decrease was observed by treatment with 50 nM narcyclasin (NCS). On the other hand, as shown in Fig. 3b, in the EGFR mutant (T790M / L858R) kinase reaction, there was no significant change in kinase activity by treatment with 50 nM gefitinib, and it was confirmed that the relative kinase activity was significantly reduced by treatment with 50 nM narcyclasin (NCS).

[0115] From the above results, it was confirmed that narcyclasin (NCS) of the present invention can inhibit or suppress the growth of cancer cells by reducing EGFR kinase activity in cancer cells with EGFR mutant type (T790M / L858R).

[0116]

[0117] Example 4. Effect of narcyclacin (NCS) on gefitinib-resistant cell lines

[0118] The PC-9 cell line, an EGFR mutant (exon 19 deletion) cell line, is a cell line that undergoes cell death due to gefitinib. Using the PC-9 cell line, gefitinib-resistant (GR) cell lines resistant to gefitinib were generated, and cell viability was confirmed by treating these cell lines with narcyclacin.

[0119] As a result, as shown in Fig. 4, both PC-9 and PC-9-GR cell lines were subjected to cell death by narcyclacin (NCS), but the number of dead cells was significantly increased in the PC-9-GR cell line, which is resistant to the tyrosine kinase inhibitor gefitinib.

[0120]

[0121] Example 5. Molecular mechanism of EGFR activation or expression reduction by narcyclasin (NCS)

[0122] 5.1. Confirmation of changes in EGFR expression due to narcyclacin (NCS) treatment

[0123] Experiments were performed to determine whether narcyclasin (NCS) affects EGFR activity in H1975 cells with EGFR mutant type (T790M / L858R).

[0124] H1975 cells were treated with 50 nM narcyclasin (NCS), and the expression levels of phosphorylated EGFR (phospho-EGFR, p-EGFR) and total EGFR were confirmed by Western blotting after 4, 8, and 24 hours. As a result, as shown in Fig. 5a, EGFR activity was significantly reduced depending on the narcyclasin (NCS) treatment time, and the expression of total EGFR was also significantly reduced at 24 hours.

[0125]

[0126] 5.2. Confirmation of the mechanism of EGFR expression changes induced by narcyclacin (NCS) treatment

[0127] To determine the cause of the decrease in EGFR expression, the expression sites of lysosomal associated membrane protein 1 (LAMP1, green), a lysosome marker that degrades intracellular substances, and EGFR (red) were confirmed through immunofluorescence (IF) analysis in H1975 cells treated with 100 nM narcyclacin (NCS).

[0128] As a result, as shown in Fig. 5b, EGFR and LAMP1 were co-stained in cell lines treated with narcyclasin (NCS). This confirmed that the decrease in EGFR expression by narcyclasin (NCS) was achieved through signal transduction related to lysosomal degradation.

[0129] In addition, to determine whether the decrease in EGFR expression by narcyclasin (NCS) is related to protein synthesis, an experiment was conducted to determine the expression level of EGFR over time after co-treatment with 50 nM narcyclasin (NCS) and cycloheximide (CHX), a protein synthesis inhibitor.

[0130] As a result, as shown in Fig. 5c, in the cell line treated with narcyclacin (NCS), the expression of EGFR decreased more significantly over time with cycloheximide (CHX) treatment compared to the control group. This confirmed that the decrease in EGFR expression by narcyclacin (NCS) was due to the promotion of EGFR protein degradation.

[0131] Next, we conducted an experiment to determine whether the EGFR expression pattern induced by narcyclacin (NCS) changed when treated with MG132 and Bafilomycin A1 (Baf A1), inhibitors of the proteasome and lysosome, respectively, which degrade intracellular proteins. As a result, as shown in Fig. 5d, it was confirmed that the decrease in EGFR expression induced by narcyclacin (NCS) was restored by treatment with a proteasome inhibitor or a lysosomal inhibitor.

[0132] From the above results, it was confirmed that narcyclacin (NCS) of the present invention can reduce the expression of EGFR by promoting the degradation of EGFR through the proteasome and lysosomal pathways.

[0133]

[0134] Example 6. In vivo anticancer effect of narcyclacin (NCS)

[0135] To confirm the in vivo anticancer effect of narcyclasin (NCS), experiments were conducted in a Caenorhabditis elegans (C. elegans) cancer model harboring the EGFR mutant (L858R / T790M). Unlike the wild-type C. elegans cancer model, the EGFR mutant (L858R / T790M) develops abnormal multivulva formation, as indicated by the red arrow in Figure 6A.

[0136] In this regard, as shown in Fig. 6B, it was confirmed that when narcyclasin (NCS) was treated in a C. elegans cancer model with an EGFR mutant (L858R / T790M), the formation of multivulva was significantly inhibited. In particular, as shown in Fig. 6C, gefitinib, a conventional first-generation EGFR kinase inhibitor, had no effect in inhibiting multivulva formation, but narcyclasin (NCS) of the present invention was confirmed to have an effect in significantly inhibiting multivulva formation.

[0137] Accordingly, it was confirmed that the narcyclasin (NCS) of the present invention has an anticancer effect in an individual with an EGFR mutant type (L858R / T790M).

[0138]

[0139] From the above results, it was found that the narcyclasin (NCS) of the present invention can significantly inhibit or suppress the growth of cancer cells by promoting the degradation of EGFR in cancers with EGFR mutations or cancers resistant to tyrosine kinase inhibitors (TKIs) and significantly reducing the activity or expression of EGFR, and thus can be usefully used for the prevention or treatment of cancer.

[0140]

[0141] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A pharmaceutical composition for preventing or treating cancer having an epidermal growth factor receptor (EGFR) mutation, comprising narcisclase or a pharmaceutically acceptable salt thereof.

2. In paragraph 1, A composition wherein the cancer is a solid cancer having an EGFR mutation.

3. In paragraph 2, A composition wherein the solid cancer is selected from the group consisting of lung cancer, breast cancer, colon cancer, colorectal cancer, pancreatic cancer, kidney cancer, head and neck cancer, ovarian cancer, cervical cancer, prostate cancer, skin cancer, liver cancer, thyroid cancer, gallbladder cancer, bile duct cancer, esophagus cancer, bladder cancer, and oral cancer.

4. In paragraph 2, A composition wherein the above solid cancer is lung cancer.

5. In paragraph 1, A composition wherein the EGFR mutation comprises at least one mutation selected from the group consisting of L858R mutation and T790M mutation.

6. In paragraph 1, The composition above is a composition that inhibits or reduces the activity or expression of EGFR.

7. In paragraph 1, The composition above promotes degradation of EGFR through the proteasomal degradation pathway or the lysosomal degradation pathway.

8. A pharmaceutical composition for preventing or treating cancer resistant to a tyrosine kinase inhibitor (TKI) comprising narcyclacin or a pharmaceutically acceptable salt thereof.

9. A food composition for preventing or improving cancer having an EGFR mutant type, comprising narcyclacin or a food-related acceptable salt thereof.

10. A food composition for preventing or improving cancer resistant to a tyrosine kinase inhibitor (TKI) comprising narcyclacin or a food-chemically acceptable salt thereof.

11. An anticancer adjuvant comprising narcyclacin or a pharmaceutically acceptable salt thereof.

12. Use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer having an EGFR mutation type.

13. Use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer resistant to tyrosine kinase inhibitors.

14. Use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer having an EGFR mutation type.

15. Use of a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer resistant to tyrosine kinase inhibitors.

16. A method for preventing or treating cancer having an EGFR mutant type, comprising administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

17. A method for preventing or treating cancer resistant to a tyrosine kinase inhibitor, comprising administering to a subject a composition comprising narcyclacin or a pharmaceutically acceptable salt thereof.

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