KR101751371B1

KR101751371B1 - Anticancer composition comprising 3,4,5-trihydroxybenzoic acid, its derivatives or its salts for preventing or treating receptor tyrosine kinase inhibitor resistant tumor

Info

Publication number: KR101751371B1
Application number: KR1020150090436A
Authority: KR
Inventors: 정양식; 김기우; 모상현
Original assignee: 연세대학교 원주산학협력단
Priority date: 2014-06-25
Filing date: 2015-06-25
Publication date: 2017-06-27
Also published as: KR20160008105A; WO2015199454A1

Abstract

The present invention relates to a trihydroxybenzoic acid (gallic acid) compound or a derivative thereof, which specifically inhibits a cancer cell that is resistant to an anticancer agent that targets a receptor tyrosine kinase The present invention relates to a composition for preventing or treating a cancerous cancer against a target anticancer agent comprising an active ingredient, wherein the trihydroxybenzoic acid (gallic acid) compound or a derivative thereof is a receptor tyrosine kinase kinase of the present invention can be prevented or treated by a receptor tyrosine kinase inhibitor such as gefitinib by specifically killing cancer cells that are resistant to an anticancer drug that targets cancer cells.

Description

TECHNICAL FIELD [0001] The present invention relates to a receptor tyrosine kinase inhibitor-resistant anticancer composition comprising 3,4,5-trihydroxybenzoic acid, a derivative thereof or a salt thereof as an active ingredient. treating receptor tyrosine kinase inhibitor resistant tumor}

The present invention relates to a trihydroxybenzoic acid (gallic acid) compound or a derivative thereof, which specifically inhibits a cancer cell that is resistant to an anticancer agent that targets a receptor tyrosine kinase And a composition for preventing or treating resistant cancer diseases against a target anticancer agent which is contained as an active ingredient.

According to a recent report by the Central Cancer Registry, cancer incidence in Korea has increased by 3.5% every year for the past 10 years. In 2010, more than 200,000 people were newly diagnosed with cancer. In Korea, the odds of getting cancer are almost two out of five, and women up to 84 years old have only one in three. Cancer is becoming increasingly common disease. The number of new cancer patients in the world was 12.7 million in 2008, 5.6 million in developed countries and 7.1 million in developing countries. By 2030, there will be 21.4 million new cancer cases and death toll of 13.2 million cancer cases. As the incidence of cancer continues to increase not only in Korea but also in the world, the importance of developing effective cancer drugs is also increasing. The global market for anticancer drugs will exceed 70 trillion won by 2010, reaching 84 trillion won by 2013. This sustained growth in market size supports the fact that research for the development of anticancer drugs is actively being carried out globally.

Conventional anticancer drugs have been used to block the cell cycle of each stage, which can induce cytotoxicity. However, this toxicity has caused side effects such as nausea, vomiting and hair loss while attacking normal cells. However, recently, in order to overcome the side effects of these conventional cytotoxic anticancer drugs, research on target anticancer drugs targeting only cancer cells has become popular. The target anticancer agent is noticed that the cancer cells are generated by the gene mutation process of the normal cells, and the agent that reacts with the mutation of the cell is targeted, and the substance that reacts with the target is administered to block the change of cancer cells . Targeted anticancer drugs have little effect on normal cells, so they are attracting attention as a next-generation anticancer drug with relatively low side effects. Therefore, investments and research are being conducted competitively to preempt the target cancer drug market worldwide.

Recently, the tyrosine kinase inhibitor (EGFR-TKIs) of the epithelial cell growth factor receptor has been attracting attention as a novel therapeutic agent for non-small cell lung cancer as a molecular target therapeutic agent (Shepherd FA et al (1994) Semin Oncol 21: 7-18).

Epidermal growth factor receptor (EGFR) is the first known receptor tyrosine kinase receptor tyrosine kinase, a membrane binding protein of 170 kilodaltons (kDa) expressed on the surface of epithelial cells. Epithelial cell growth factor receptors are members of the growth factor receptor family of the protein tyrosine kinases, the cell cycle regulatory molecule class (W. J. Gullick et al., 1986, Cancer Res., 46: 285-292). The epithelial growth factor receptor is activated when its ligand epithelial cell growth factor (EGF) binds to the extracellular domain, thereby causing autophosphorylation of the intracellular tyrosine kinase domain of the receptor (S. Cohen et al., 1980 , J. Biol. Chem., 255: 4834-4842; AB Schreiber et al., 1983, J. Biol. Chem., 258: 846-853) signal transduction pathway is activated and cell proliferation occurs as a result.

The epithelial growth factor receptor is a protein product of erbB or ErbB1, a growth-promoting oncogene, which is a member of the ERBB family of primary oncogenes and is believed to play a pivotal role in the pathogenesis and progression of many human cancers. Indeed, the expression of the receptor is continuously observed in overexpressed or mutant forms of epithelial growth factor receptor in breast cancer, bladder cancer, lung cancer, head cancer, cervical cancer, gastric cancer as well as glioblastoma.

Cancer occurs because cell proliferation is not regulated as a result of genetic mutations affecting the function of growth factor receptor function or inducing overexpression of the receptor and / or ligand.

In particular, the epithelial growth factor receptor has been shown to play an important role in the resistance of cancer cells to cell differentiation, cell motility enhancement, protein secretion, angiogenesis, invasion, metastasis, chemotherapeutic agents and radiation (MJ Oh et al , 2000, Clin. Cancer Res., 6: 4760-4763).

In addition to the hypothesis that suppression of epithelial growth factor receptor may be a target of new chemotherapy, monoclonal antibodies that inhibit receptor binding of the ligand by its therapeutic method and small molecules inhibiting autophosphorylation of tyrosine kinase of intracellular domain (Kawamoto et al (1983) Proc. Natl. Acad Sci. USA 80: 1337-1341).

Thus, epithelial growth factor receptors have been the target of choice for the development of several different cancer therapies. EGFR tyrosine kinase inhibitors (EGFR-TKIs) are one of these therapies because activation of the epithelial growth factor receptor pathway requires reversible phosphorylation of tyrosine residues. In other words, epithelial growth factor receptor tyrosine kinase inhibitors (EGFR-TKI) block the activity of cell surface receptors that are responsible for triggering and / or maintaining signaling pathways in cells that cause growth and differentiation of tumor cells. Specifically, these inhibitors block the activity of the receptor by blocking the adenosine triphosphate (ATP) binding domain of the epithelial cell growth factor receptor kinase, termed HER-1, so that the ATP does not bind any more . The epithelial growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) is known as the quinazolines pyridopyrimidines, and pyrrolopyrimidines compounds.

Recently, a variety of inhibitors against epithelial growth factor receptor have been identified and two new quinazolines compounds approved after clinical development have been identified, including Gefitinib (compound ZD1839 developed by AstraZeneca UK Ltd.; (Available under the trade name IRESSA) and Erlotinib (the compound OSI774 developed by Genentech, Inc. and OSIPharmaceuticals, Inc., available under the trade name TARCEVA), both of which exhibited encouraging clinical results Presentations at ASCO 2003 and WCLC, Aug.2003).

In particular, Gefitinib is the first commercially available orally administered epithelial growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) and has been shown to inhibit the growth of various solid tumors such as lung, colon, and breast cancer in preclinical studies William Pao et al (2005) J Clin Oncol 23: 2556-2568; Herbst R et al (2004) Nat Rev Cancer 4: 956-65; Miller VA et al (2004) J Clin Oncol 22: 1103-1109). Gefinitib is a small synthetic anilinoquinazoline that has minimal inhibitory activity on other tyrosine kinases (TKs) and selectively activates the epithelial growth factor receptor tyrosine kinase (EGFR-TK) . It also inhibits mitogen-activated protein kinase (MAPK) and inhibits the subsequent nuclear transcription factor c-fos. Basically, this drug inhibits growth but induces apoptosis at high doses. Gefitinib has not only improved the toxicity and side effects of conventional cytotoxic anticancer drugs such as bone marrow suppression, nephrotoxicity, nausea, vomiting and alopecia but also oral treatment.

However, the epithelial growth factor receptor (EGFR) target inhibitor, gefitinib, has been implicated in the development of drug resistance due to the expression of the epithelial growth factor receptor (EGFR) T790M or the protooncogene c-Met Respectively. In fact, it has been shown that zetytipine has a remarkable therapeutic effect with a response rate of about 15% in patients. However, when taken for about 1 year, it is important to make a plan to overcome the cancer, It is emerging as a task.

Trihydroxybenzoic acid, also called gallic acid or gallate, is found in various plant extracts. (1999) Anticancer Drugs 10 (9): 845-51; Ho et al. (1999) have shown that gallic acid inhibits the growth of cancer cells such as lung cancer, stomach cancer and uterine cancer and induces apoptosis. (2010) Food and Chemical Toxicology 48 (8-9): 4834-4842; Zhao et al. (2013) Onco Letters 6: 1749-1755).

However, the anticancer effect that specifically kills cancer cells that are resistant to epithelial growth factor receptor inhibitors such as zetitifib and / or erlotinib has not been disclosed.

U.S. Patent No. 8,334,000 discloses that gallic acid and various derivatives of gallic acid contained in extracts of other plants including pomegranate inhibit the growth of malignant tumors by inhibiting angiogenesis, There is no mention of the therapeutic effect on cancers resistant to tyrosine kinase inhibitors at all.

Korean Patent Laid-Open Publication No. 2014-0015798 discloses a daphnane diterpenoid-based substance as an anticancer drug that kills cancer cells that are resistant to the Gefitinib anticancer drug. However, It is believed that a long validation period will be required until it is actually used as a resistance agent for the currently marketed zetotinib, since it kills not only cancer cells but also cancer cells which do not show resistance.

Korean Patent Publication No. 2007-0107693 (International Publication No. WO 2006/084058) discloses the use of EKB-569, HKI-272 or HKI-272 as an irreversible epithelial growth factor receptor antagonist against sextetin in zetitib or erlotinib, 357. However, this is based on the second mutation T790M occurring in the kinase domain among endometrial cancer cases, and the second mutation does not occur in all endometrial cancer, and since the compound is not a naturally occurring component, it is relatively toxic Therefore, a long verification time will be required until it is actually used as a resistance agent of zeta-nip.

Therefore, it selectively acts on cancer cells that are resistant to epithelial growth factor receptor inhibitors such as zetitib and / or erlotinib, which are currently on the market, to selectively kill resistant cancerous cells, A naturally occurring compound that can maximize the effect and minimize side effects is required.

United States Patent No. 8,334,000 Korean Patent Laid-Open Publication No. 2014-0015798 Korean Patent Publication No. 2007-0107693 (International Publication No. WO 2006/084058)

Shepherd FA et al., 1994, Semin Oncol., 21: 7-18 W. J. Gullick et al., 1986, Cancer Res., 46: 285-292 S. Cohen et al., 1980, J. Biol. Chem., 255: 4834-4842; A. B. Schreiber et al., 1983, J. Biol. Chem., 258: 846-853 M. J. Oh et al., 2000, Clin. Cancer Res., 6: 4760-4763 Kawamoto et al., 1983, Proc. Natl. Acad. Sci. USA, 80: 1337-1341 William Pao et al., 2005, J Clin Oncol., 23: 2556-2568 Herbst R et al., 2004, Nat. Rev. Cancer, 4: 956-65 Miller VA et al., 2004, J. Clin. Oncol., 22: 1103-1109 Ohno Y et. al., 1999, Anticancer Drugs, 10 (9): 845-51 Zhao et al., 2013, Oncology Letters 6: 1749-1755 C. Locatelli et al., 2012, Euro. J. of Medi. Chem., 60: 233-239

It is an object of the present invention to provide a composition capable of inducing intracellular toxicity or apoptosis of cancer cells showing tolerance to an anticancer drug targeting receptor tyrosine kinase.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or ameliorating tolerable cancer diseases against a target anticancer drug comprising 3,4H-trihydroxybenzoic acid compound, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient. A composition for treating or preventing cancer, a functional anticancer agent, a functional health food capable of preventing or ameliorating cancer disease, a food which can prevent and improve cancer, including the effective ingredient, and an anticancer agent, To provide a combination that is capable of preventing or treating a target cancer-resistant cancer disease.

The composition according to the present invention specifically inhibits the receptor tyrosine kinase (receptor tyrosine kinase) -receptor-resistant cancer diseases such as gefitinib by specifically killing cancer cells that are resistant to an anticancer drug targeting receptor tyrosine kinase Can be prevented or treated.

Figure 1 shows the changes in cell growth of HCC827 cell line and resistant HCC827GR (gefitinib resistnace) cell line without resistance to gefitinib following treatment with different concentration of gefitinib by MTT assay As shown in FIG.
FIG. 2A shows the cell growth of HCC827 cell line and resistant HCC827GR (Gefitinib Resistnace) cell line which are not resistant to gefitinib following treatment with 3,4hydroxybenzoic acid (gallic acid), 3,4,5-trihydroxybenzoic acid (MTT) analysis of the change.
FIG. 2b is a graph showing the results of the reaction of 3,4-trihydroxybenzoic acid, gallic acid, gefitinib, 4-hydroxy-3-methoxybenzoic acid, vanillic acid ) And the colony formation assay of HCC827 cells and resistant HCC827GR (Gefitinib Resistnace) cells without resistance to gefitinib following treatment of each vehicle (Vehicle, Veh) .
Figure 3a shows the sensitivity of the cell lines of H358, H1650, H1666, H1734, H1975, HCC827, and H3255 to the expression of gefitinib and 3,4hydroxylbenzoic acid (gallic acid) (MTT) analysis method.
Figure 3b is a graph showing the effect of each treatment of 3,4,8-trihydroxybenzoic acid (gallic acid), gefitinib, and vehicle (Veh) on gefitinib The results of the colony formation assay of the resistant H358 and H1975 cell lines are shown.
Figs. 4A and 4B are graphs and photographs showing the results of a mouse experiment, showing a decrease in tumor volume. Fig.

The present invention relates to a method for the prevention of cancer-resistant diseases against target anticancer drugs comprising 3,4-trihydroxybenzoic acid compound represented by the following formula (1), a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient Or therapeutic compositions.

[Chemical Formula 1]

.

The present invention relates to a pharmaceutical composition comprising a trihydroxybenzoic acid compound represented by the following Formula 1, which is found in various plant extracts, as an epithelial growth factor receptor (EGFR) it has been confirmed that 3,4,5-trihydroxybenzoic acid compound is useful for the treatment or prevention of cancerous cancer resistant to the anticancer drug by confirming that it specifically reacts with cancer cells showing resistance to gefitinib anticancer drug and killing it .

[Chemical Formula 1]

.

In one embodiment according to the present invention, the compound, its derivative or a pharmaceutically acceptable salt thereof may exhibit cytotoxicity in a cancer cell having resistance to a receptor tyrosine kinase target anticancer drug.

In one embodiment according to the present invention, the compound, its derivative or pharmaceutically acceptable salt thereof may exhibit cytotoxicity in a cancer cell which is resistant to an epithelial growth factor receptor (EGFR) target anticancer drug.

In one embodiment according to the present invention, the compound, a derivative thereof or a pharmaceutically acceptable salt thereof may exhibit a higher cytotoxic inducing effect in a drug-resistant cancer cell than in a normal cancer cell.

In one embodiment according to the present invention the targeted anticancer agent is selected from the group consisting of Gefitinib, temozolomide, bevacizumab, erlotinib, capecitabine and imatinib imatinib), but the present invention is not limited thereto.

In one embodiment according to the present invention the cancer disease is selected from the group consisting of breast cancer, lung cancer, non-small cell lung cancer, gastric cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, head cancer, skin cancer, malignant melanoma, uterine cancer, Endometrioid carcinoma, endometrioid carcinoma, vulvar carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small bowel cancer, lymphadenocarcinoma, bladder cancer, endometrioid carcinoma, endometrioid carcinoma, Neoplasms, renal cell carcinomas, renal pelvic carcinomas, central nervous system (CNS) neoplasms, papillary carcinomas, pancreatic carcinomas, pancreatic carcinomas, pancreatic carcinomas, central nervous system (GI) tumors, primary central nervous system lymphoma, spinal cord tumors, brainstem glioma, and pituitary adenoma.

In one embodiment according to the present invention, the composition may be used alone or in combination with other anticancer agents.

In one embodiment according to the present invention, the compounds of the present invention or derivatives thereof may be prepared as pharmaceutically acceptable salts and solvates according to methods conventional in the art.

In one embodiment according to the present invention, as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts may be prepared in a conventional manner, for example, by dissolving the compound in an excess of an acid aqueous solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile, The acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be filtered by suction.

In one embodiment, the free acid may be an organic acid or an inorganic acid. Examples of the inorganic acid include hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like. Examples of the organic acid include methanesulfonic acid, p-toluenesulfonic acid, But are not limited to, acetic, trifluoroacetic, citric, maleic, succinic, oxalic, benzoic, tartaric, fumaric, mandelic, propionic, citric, glycollic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, But is not limited thereto.

In one embodiment according to the invention, the base can be used to make a pharmaceutically acceptable metal salt. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable to produce sodium, potassium or calcium salt particularly as a metal salt, and the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

In one embodiment according to the present invention, the pharmaceutically acceptable salts of the compounds of the invention or derivatives thereof may, unless otherwise indicated, include salts of acidic or basic groups which may be present in the compounds, Pharmaceutically acceptable salts include the sodium, calcium and potassium salts of the hydroxy group and the other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, (Mesylate) and p-toluenesulfonate (tosylate) salts, which may be prepared by methods or processes known to those skilled in the art, such as, for example, .

The present invention relates to a pharmaceutical composition for prevention or treatment of cancer-resistant cancer, which comprises as an active ingredient 3,4,5-trihydroxybenzoic acid compound represented by the following formula (1), a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient Cancer adjuvant.

[Chemical Formula 1]

.

In one embodiment of the present invention, the compound, its derivative or a pharmaceutically acceptable salt thereof may exhibit cytotoxicity in a cancer cell having resistance to a receptor tyrosine kinase target anticancer drug.

In one embodiment according to the present invention, the compound, its derivative or a pharmaceutically acceptable salt thereof may exhibit cytotoxicity in a cancer cell which is resistant to the epithelial growth factor receptor (EGFR) target anticancer drug.

In one embodiment of the present invention, the anticancer adjuvant of the present invention may further contain one or more active ingredients exhibiting the same or similar functions, and the pharmaceutically acceptable carrier may be saline, sterilized water, Ringer's solution, buffer Saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and at least one of these components may be used in combination.

The present invention relates to a pharmaceutical composition for prevention of cancer-resistant cancer diseases, which comprises as an active ingredient 3,4,5-trihydroxybenzoic acid compound represented by the following formula (1), a derivative thereof or a pharmaceutically acceptable salt thereof and an anti- Or < / RTI > therapeutic combinations.

[Chemical Formula 1]

.

In one embodiment of the present invention, the anticancer agent includes antimetabolites such as methotrexate, 6-mercaptopurine, 6-thioguanine, 5-fluorouracil and cytarabine; An alkylating agent such as a chlorambucil, a cyclophosphamide, an ethylene imine compound, an alkyl sulfonate compound, a carnustine compound or a dacarbazine compound, ); Antitumor agents such as actinomycin D, doxorubicin, bleomycin and mitomycin, plant alkaloids such as vincristine and vinblastine, and mitotic inhibitors such as taxoid, a mitotic inhibitor containing a taxane ring antimitotic drugs); Hormones such as corticosteroids or progesterone; Platinum-containing compounds such as cisplatin, but are not limited thereto.

The present invention includes the step of administering an effective amount of said composition, anticancer adjuvant, or combination thereof to a patient in need of gametin treatment of zetitib or erlotinib.

In one embodiment according to the present invention, the treatment may include treatment with other receptor tyrosine kinases such as zetitib or erlotinib, chemotherapy, radiation therapy, and the like, It is not.

In one embodiment according to the present invention, an effective amount of said composition, anticancer adjuvant, or combination thereof may be administered to a mammal, including a human, by a variety of routes.

In one embodiment according to the present invention, the effective amount may vary depending on the age, weight, sex, health condition, drug form, dosage form, disease severity, tumor mass, A statistically significant fraction can be determined taking into account the amount that yields a beneficial effect.

In one embodiment according to the present invention, the effective dose will generally be from 0.001 mg / kg to 10 g / kg per day, more preferably from 0.01 mg / kg to 1 g / kg per day, And may be administered in divided doses as needed, but the dose is not intended to limit the scope of the invention in any way.

In one embodiment according to the present invention, in addition to the active ingredient of the present invention, one or more further pharmaceutically acceptable carriers may be prepared.

In one embodiment according to the present invention, the pharmaceutically acceptable carrier is selected from the group consisting of saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, And other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added as needed. In addition, it can be formulated into injection formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like by additionally adding diluents, dispersants, surfactants, binders and lubricants, Specific antibody or other ligand can be used in combination with the carrier. Can further be suitably formulated according to the respective disease or ingredient, using appropriate methods in the art or as disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA have.

In one embodiment according to the present invention, the formulations may be prepared using, but not limited to, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like.

In one embodiment according to the present invention, the dosage form is not particularly limited, but may be administered parenterally or orally, as desired.

In one embodiment according to the present invention said parenteral administration is intravenous (iv), intramuscular (im), subcutaneous (sc), intradermal (id), intraperitoneal (ip), intrathecal (it) Intramuscular, intrathecal, intrauterine, rectal, vaginal, topical, intratumoral, and the like, and may be administered parenterally by injection or by gradual infusion over time and delivered by peristaltic means.

In one embodiment of the present invention, the preparation for parenteral administration includes a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried preparation, a suppository, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

In one embodiment according to the present invention, the solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, troches and the like, which may contain one or more excipients For example, it is prepared by mixing starch, calcium carbonate, sucrose, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions or syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have.

In one embodiment according to the present invention, the agent for oral administration may be provided together with an enhancer. Orally acceptable absorption enhancers include surfactants such as sodium lauryl sulfate, palmitoyl carnitine, laureth-9, phosphatidylcholine, cyclodextrin and their derivatives; Bile salts such as sodium deoxycholate, sodium taurocholate, sodium glycocholate, and sodium pushidate; Chelating agents such as EDTA, citric acid and salicylate; And fatty acids (e.g., oleic acid, lauric acid, acylcarnitines, mono and diglycerides). Other oral absorption enhancers include benzalkonium chloride, benzethonium chloride, CHAPS (3- (3-cholamidopropyl) -dimethylammonio-1-propanesulfonate), Big-CHAPS (N, 3-D-glucoamidopropyl) -colamide), chlorobutanol, octoxynol-9, benzyl alcohol, phenols, cresols, and alkyl alcohols.

In one embodiment according to the present invention, the active ingredient of the present invention may be administered in liposomes or microspheres (or microparticles).

In one embodiment according to the present invention, the active ingredient of the present invention can be implanted into a complex of microspheres or microspheres and transplanted to be slowly released over a period of several days to several months.

In one embodiment according to the present invention, the microspheres are formed from polymers or proteins and are known to those skilled in the art and can be tailored to pass through the gastrointestinal tract directly into the bloodstream.

In one embodiment according to the present invention, the active ingredient of the present invention may be formulated into the liposomes or microparticles (microspheres) and administered intravenously in a size suitable to lodge capillary blood vessels.

In one embodiment according to the present invention, when the liposomes or microparticles reach capillary blood vessels around the ischemic tissue, the active ingredients may be administered topically to the site where they may be most effective.

In one embodiment according to the present invention, the liposome is generally less than about 200 nm and may be a multilayer vesicle to target ischemic tissue.

In one embodiment according to the present invention, the microparticles may be those made from biodegradable polymers such as polyglycolide, polylactide and copolymers thereof, and those skilled in the art will appreciate that the desired drug release rate and the desired dosage A suitable carrier system can be easily determined according to various factors such as the amount of the carrier.

In one embodiment according to the present invention, the formulation containing the active ingredient of the present invention may be administered directly into a blood vessel through a catheter, wherein the active ingredient may be contained in the biodegradable polymer hydrogel.

In one embodiment according to the present invention, the polymer hydrogel may be delivered into a tissue lumen, and a formulation containing the active ingredient of the present invention may be released over time as the polymer degrades.

In one embodiment according to the present invention, the polymeric hydrogel may have microparticles or liposomes in which the active ingredient of the present invention is dispersed, which provides another mechanism for controlled release of the active ingredient of the present invention .

In one embodiment according to the present invention, the agent comprising the active ingredient of the present invention for such administration may be prepared by a method known in the pharmaceutical field and suitably provided in unit dosage form, Contacting the component with a carrier that constitutes one or more accessory ingredients.

In one embodiment according to the present invention, the preparation is prepared by uniformly and densely contacting the active ingredient of the present invention with a liquid carrier or a finely divided solid carrier, and then, if necessary, shaping the product into the desired unit dosage form ).

In one embodiment according to the present invention, the formulation comprising the active ingredient may contain one or more optional auxiliary ingredients such as diluents, buffers, spices, binders, surface active agents, thickeners, lubricants, suspensions A preservative (antioxidant, etc.), and the like.

In one embodiment according to the present invention, the active ingredient of the present invention may be administered alone or in combination with an inert carrier such as lactose, for administration to the respiratory tract, as a solution for snuff or an aerosol or inhaler or as an ultra- In which case the particles of the active ingredient are suitable to have a diameter of less than 50 microns, preferably less than 10 microns, more preferably 2 to 5 microns.

In one embodiment according to the present invention, the composition for nasal administration is desirably weakly acidic in pH, and more specifically the pH is preferably from about 3 to about 5, more specifically from about 3.5 to about 3.9, Hydrochloric acid, and the like.

The present invention relates to a pharmaceutical composition for preventing or ameliorating a cancer-resistant cancer drug, which comprises a 3,4-trihydroxybenzoic acid compound represented by the following formula (1), a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient Health functional foods.

[Chemical Formula 1]

.

The health functional food may be variously used for medicines, foods, beverages and the like, and can be commercialized in the form of tablets, capsules, pills, powders, granules, liquid beverages, concentrates and the like.

In the case of health functional foods in the form of granules, tablets or capsules, it is generally 0.0001 to less than 100% by weight, more particularly 0.01 to 80% by weight, more particularly 0.1 to 50% by weight, more particularly 1 to 20% The active ingredient of the present invention may be added, but not limited thereto.

The present invention relates to a food for preventing and / or improving cancer, which comprises as an active ingredient 3,4,5-trihydroxybenzoic acid compound represented by the following formula (1), a derivative thereof or a pharmaceutically acceptable salt thereof .

[Chemical Formula 1]

.

Such foods include beverages, meats, chocolates, confectionery, pizza. Examples of the foodstuffs include noodles, noodles, gums, ice creams, vitamin complexes, alcoholic beverages, dairy products, juices, rings, granules, teabags and sticks.

Generally, the food composition of the present invention may be added in an amount of 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

In one embodiment according to the present invention, the health beverage composition of the present invention is not limited to liquid ingredients other than containing the active ingredient mixture of the present invention as an essential ingredient in the indicated ratios, and includes various flavors Or natural carbohydrate as an additional ingredient. Examples of such natural carbohydrates include conventional saccharides such as monosaccharides such as disaccharides such as glucose and fructose such as maltose and sucrose and polysaccharides such as dextrin and cyclodextrin and xylitol such as sorbitol , Erythritol, and the like. As a flavor other than the above, a natural flavoring agent (tau Martin, a stevia compound (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavorings (e.g., saccharin, aspartame etc.) The ratio of the natural carbohydrate is generally about 1 to 20 g, more specifically about 5 to 12 g per 100 ml of the beverage composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following Examples, Experimental Examples and Preparation Examples. It should be understood, however, that the same is by way of illustration and example only and is not intended to limit the scope of the present invention.

[Experimental Example]

<3,4,5-Trihydroxybenzoic acid compound>

The 3,4, 5-trihydroxybenzoic acid compound used in the experimental examples was purchased from Sigma-Aldrich and Alfa Aesar.

Where to buy Product Number (Catalog #) Sigma-Aldrich G7384 Alfa Aesar B24887

Experimental Example 1: Gefitinib-resistant cell line

HCC827GR # 1 and HCC827GR # 2 cell lines resistant to HCC827 and zetti nip were inoculated into 96-well plates, respectively. To measure the cell growth response to zetitifib, zetitipib was treated with 0.1, 0.3 , 1, 3, and 5 μM, respectively. After 5 days, cell viability was measured by MTT assay to confirm the growth of the cells. As a result, as shown in Fig. 1, resistance to zetotinib was high in the order of HCC827 <HCC827GR # 1 <HCC827GR # 2.

Experimental Example 2 Cell Growth Response to 3,4 Hydroxybenzoic Acid of HCC827 Cell Lines

HCC827GR # 1 and HCC827GR # 2 cell lines resistant to HCC827 and zetitibe were inoculated in 96-well plates, respectively, in order to measure the cell growth reaction against 3,4,5-trihydroxybenzoic acid, Twenty-four hours after the inoculation, 3,4,5-trihydroxybenzoic acid was treated at the concentrations of 0, 5, 10, 25, 35 and 50 uM, respectively. After 5 days, MTT assay was used to measure cell viability ) Was measured to confirm the growth reaction of the cells. As a result, as shown in Fig. 2a, HCC827GR # 2 cell line which is most resistant to gefitinib was treated with 3,4H-trihydroxybenzoic acid (gallic acid) And it was confirmed that it was sensitively inhibited.

HCC827GR # 1 and HCC827GR # 2 cell lines resistant to HCC827 and gefitinib were measured on 6-well plates And after 24 hours of inoculation, 0.3 uM zetitibim, 25 uM 3,4,5-trihydroxybenzoic acid, and 25 uM 4-hydroxy-3-methoxybenzoic acid, vanillic acid) and then 10 days later, cell viability was measured by colony formation assay using methylene blue staining to confirm the growth of the cells.

As a result, as shown in FIG. 2B, in the colony formation assay, cell growth inhibition and clonogenicity due to treatment with 3,4,5-trihydroxybenzoic acid were found to be dependent on the degree of resistance to zetti nip I could confirm the proportion. Furthermore, in the case of 4-hydroxy-3-methoxybenzoic acid, which is a similar polyphenol, no anticancer effect was observed. These results indicate that the antitumor effect of 3,4,5-trihydroxybenzoic acid is more specific to the sex pheromone of polyphenols than other polyphenols.

Experimental Example 3: Trihydroxybenzoic acid anti-cancer effect test from various lung cancer cell lines

Table 2 below shows IC ₅₀ values for gefitinib in various lung cancer cell line panels with K-ras mutation and epithelial growth factor receptor (EGFR) mutation characteristics. (Table 2: Lt; RTI ID = 0.0 > IC50 < _{/ RTI} > values for gefitinib)

HCC827GR # 1 and HCC827GR # 2 cell lines resistant to H358, H1975, H1650, HCC827, H1666, H3255 and zetitipine listed in Table 2 were each inoculated into 6-well plates, and after 24 hours of inoculation, Hydroxybenzoic acid, vanillic acid, and 25 uM of 3,4-trihydroxybenzoic acid, respectively. After 5 days, The cell viability was measured by the MTT assay and the growth reaction of the cells was confirmed.

As a result, as shown in FIG. 3A, treatment of various lung cancer cell lines with 3,4,5-trihydroxybenzoic acid (gallic acid) resulted in a decrease in the sensitivities of japitinib and 3,4,5-trihydroxybenzoic acid As a result, the sensitivities of zetipine and 3,4,5-trihydroxybenzoic acid were found to be inversely proportional to almost all different lung cancer cell lines.

In addition, in order to measure the growth reaction of 3,4,5-trihydroxybenzoic acid in two cell lines having different mutations, but commonly having resistance to zetti nip, H358 having K-ras mutation and epithelium H1975 cell lines harboring mutants of the cell growth factor receptor (EGFR) were each inoculated into 6-well plates, and after 24 hours of inoculation, 0.3 uM zetitibim, 25 uM 3,4,5-trihydroxybenzoic acid, and 25 uM 4- Hydroxy-3-methoxybenzoic acid and vanillic acid, respectively. After 10 days, the cells were treated with a colony formation assay using methylene blue staining to determine cell viability (cell viability) was measured to confirm the growth reaction of the cells.

As a result, clonogenicity was disappeared by treatment with 3,4,5-trihydroxybenzoic acid in both H358 and H1975 cell lines, as shown in Fig. 3B. Therefore, it was confirmed that 3,4,5-trihydroxybenzoic acid had a specific anticancer effect if it had a zetti-nip resistance even in a mutant lung cancer cell line.

Experimental Example 4. Animal Experiment (tumor growth inhibitory effect)

A xenotransplantation experiment was conducted to determine the inhibitory effect of 3,4,5-trihydroxybenzoic acid on tumor growth. The experimental method is as follows:

5 × 10 ⁶ H1650 lung cancer cells were subcutaneously injected into the side of nude mice. H1650-transplanted nude mice were randomly divided into two groups, four as control and six as experimental groups. DMSO was intraperitoneally administered to the control mice for 32 days from the time when the tumor volume reached 20 mm ³ , and the dose of 200 mg / kg of 3,4,5-trihydroxybenzoic acid was administered to the experimental mice every 2 days. The tumor volume was calculated as (x width x length x height).

Experimental results showed that the experimental group treated with 3,4,5-trihydroxybenzoic acid significantly inhibited tumor growth compared with the control group treated with DMSO. 4A is a graph showing the tumor growth inhibitory effect of H1650 transplanted mice. (p = 0.0211) It can be seen that when the 3,4,5-trihydroxybenzoic acid according to the present invention is administered, the tumor size is significantly reduced.

[Manufacturing Example]

Production Example 1. Preparation of powder

Trihydroxybenzoic acid (gallic acid) 200 mg

Lactose ------------------------------------------------- - 100 mg

Talc ------------------------------------------------- - 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Production Example 2. Preparation of tablets

Trihydroxybenzoic acid (gallic acid) 200 mg

Corn starch --------------------------------------------- 100 mg

Lactose ------------------------------------------------- - 100 mg

Magnesium stearate ------------------------------------ 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Preparation Example 3. Preparation of capsules

Trihydroxybenzoic acid (gallic acid) 200 mg

Crystalline cellulose - 3 mg

Lactose ---------------------------------------------- 14.8 mg

Magnesium stearate 0.2 mg < RTI ID = 0.0 >

The above components are mixed in accordance with a conventional method for producing a capsule, and filled in a gelatin capsule to prepare a capsule.

Production Example 4. Preparation of injection

Trihydroxybenzoic acid (gallic acid) 200 mg

Mannitol ------------------------------------------------ 180 mg

Sterile sterile distilled water for injection 1594 mg

Na ₂ HPO ₄ , 12H ₂ O ----------------------------------------- 26 mg

(2 mL) per 1 ampoule in accordance with the usual injection preparation method.

Production Example 5. Production of liquid agent

Trihydroxybenzoic acid (gallic acid) 200 mg

Isolation Party ---------------------------------------------- 10 g

Mannitol ------------------------------------------------ 5 g

Purified water ------------------------------------------------

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Production Example 6. Production of Health Food

Trihydroxybenzoic acid (gallic acid) 1000 mg

Vitamin mixture -------------------------------------------

Vitamin A Acetate ------------------------------------- 70 g

Vitamin E ---------------------------------------------- 1.0 mg

Vitamin B1 --------------------------------------------- 0.13 mg

Vitamin B2 --------------------------------------------- 0.15 mg

Vitamin B6 --------------------------------------------- 0.5 mg

Vitamin B12 -------------------------------------------- 0.2 g

Vitamin C ----------------------------------------------- 10 mg

Biotin ------------------------------------------------- 10 g

Nicotinic acid amide 1.7 mg

Folic acid ------------------------------------------------- - 50 g

Calcium pantothenate ----------------------------------------- 0.5 mg

Inorganic mixture --------------------------------------------------------------------------

Ferrous sulfate ------------------------------------- 1.75 mg

Zinc oxide ---------------------------------------------- 0.82 mg

Magnesium carbonate ------------------------------------------ 25.3 mg

Potassium phosphate monohydrate 15 mg

Dicalcium phosphate ------------------------------------------- 55 mg

Potassium citrate -------------------------------------------- 90 mg

Calcium carbonate ---------------------------------------------- 100 mg

Magnesium chloride ------------------------------------------ 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Production Example 7. Preparation of health drinks

Trihydroxybenzoic acid (gallic acid) 1000 mg

Citric acid ------------------------------------------------ 1000 mg

Oligosaccharides ---------------------------------------------- 100 g

Plum concentrate -------------------------------------------- 2 g

Taurine ------------------------------------------------ 1 g

Add purified water ---------------------------------- Total 900 mL

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

Claims

A receptor tyrosine kinase target anticancer agent including Gefitinib (Gefitinib), which is an effective ingredient, as the active ingredient, a 3,4-trihydroxybenzoic acid compound represented by the formula (1) or a pharmaceutically acceptable salt thereof, ) Or erlotinib for the prevention or treatment of resistant cancer:
[Chemical Formula 1]

.

delete

The method according to claim 1,
Wherein said compound or a pharmaceutically acceptable salt thereof is cytotoxic in a cancer cell having resistance to an epithelial growth factor receptor (EGFR) target anticancer drug. Description: BACKGROUND OF THE INVENTION 1. Field of the Invention < RTI ID = 0.0 > A pharmaceutical composition for preventing or treating resistant cancer.

The method according to claim 1,
Wherein said compound or a pharmaceutically acceptable salt thereof exhibits a higher apoptosis inducing effect than a normal cancer cell in drug resistant cancer cells. The present invention provides a method for preventing or treating resistance to zotitnib or erotinib, which is a receptor tyrosine kinase target anticancer drug, A pharmaceutical composition.

delete

The method according to claim 1,
The cancer may be selected from the group consisting of breast cancer, lung cancer, non-small cell lung cancer, gastric cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, head cancer, skin cancer, malignant melanoma, uterine cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, lymphadenocarcinoma, bladder cancer, gallbladder cancer, endocrine cancer, thyroid cancer, pituitary cancer, soft tissue sarcoma, soft tissue sarcoma, endometrioid carcinoma, endometrial carcinoma, endometrial carcinoma, uterine cancer, vulvar carcinoma, vulvar carcinoma, Cancer of the urethra, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system (CNS) neoplasm, cancer of the urethra, cancer of the urethra, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, Lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. The present invention relates to a method for the prevention or treatment of resistant cancers against zetotinib or erlotinib, which are receptor tyrosine kinase target anticancer drugs For the pharmaceutical composition.

The method according to claim 1,
Wherein the composition is used alone or in combination with other anticancer agents. The pharmaceutical composition for preventing or treating resistance to zetitinib or erotinib, which is a receptor tyrosine kinase target anticancer agent.

Receptor tyrosine kinase comprising trihydroxybenzoic acid compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient Resistance to chemotherapeutic agent zetitipine or erotinib Anticancer agents for prevention or treatment of cancer:
[Chemical Formula 1]

.

delete

9. The method of claim 8,
Wherein said compound or pharmaceutically acceptable salt thereof is a receptor tyrosine kinase which is characterized by exhibiting cytotoxicity in a cancer cell having resistance to an epithelial growth factor receptor (EGFR) target anticancer drug. Cancer adjuvant for the prevention or treatment of cancer.

9. The method of claim 8,
Wherein said compound or a pharmaceutically acceptable salt thereof exhibits a higher apoptosis inducing effect than a normal cancer cell in drug resistant cancer cells. The present invention provides a method for preventing or treating resistance to zotitnib or erotinib, which is a receptor tyrosine kinase target anticancer drug, Cancer adjuvant for.

delete

9. The method of claim 8,
The cancer may be selected from the group consisting of breast cancer, lung cancer, non-small cell lung cancer, gastric cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, head cancer, skin cancer, malignant melanoma, uterine cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, lymphadenocarcinoma, bladder cancer, gallbladder cancer, endocrine cancer, thyroid cancer, pituitary cancer, soft tissue sarcoma, soft tissue sarcoma, endometrioid carcinoma, endometrial carcinoma, endometrial carcinoma, uterine cancer, vulvar carcinoma, vulvar carcinoma, Cancer of the urethra, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system (CNS) neoplasm, cancer of the urethra, cancer of the urethra, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, Lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. The present invention relates to a method for the prevention or treatment of resistant cancers against zetotinib or erlotinib, which are receptor tyrosine kinase target anticancer drugs Cancer supplements.

delete

Receptor tyrosine kinase comprising trihydroxybenzoic acid compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient Resistance to chemotherapeutic agent zetitipine or erotinib Health food for prevention or improvement:
[Chemical Formula 1]

.

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