KR20130134666A - A composition comprising known compounds isolated from the extract of pine tree leaf for the prevention and treatment of cancer diseases - Google Patents

Abstract

The present invention relates to a compound isolated from the extract of pine needle and a composition including the same as an active ingredient. Specifically, the present invention can be useful in pharmaceutical composition and health food for preventing and treating various cancer diseases because the anticancer activity of the present invention is confirmed by checking cytotoxicity against cell lines of lung cancer, ovarian cancer, skin cancer, colon cancer, cervical cancer.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing and treating cancers containing a known compound isolated from pine needle extract,

The present invention provides a composition for preventing and treating cancer diseases, which comprises a compound isolated from pine needle extract having cytotoxicity against various cancer cell lines.

Kalechman et al., Synergistic anti-tumoral effect of paclitaxel (Taxol) + AS101 in a murine model of B16 melanoma: association with ras-dependent signal-trasduction pathways. Int . J. Cancer , 86 , pp. 281-288, 2000.

[Reference 2] Gamet-Payractre et al ., Sulforaphane a naturally occurring isothiocyanate, induced arrest and apoptosis in HT29 human colon cancer cell. Cancer Res . , 60 , pp. 1426-1433, 2000),

[Reference 3] Piao et al ., Induction of G (2) / M phase arrest and apoptosis by a new synthetic anti-cancer agent, DW2282, in promyelocytic leukemia (HL-60) cells. Bio chem Pharmacol , 62 , pp. 1439-1447, 2001).

Vanchieri, C., IARC Publishes Data on Worldwide Cancer Cases, Journal of the National Cancer Institute, 85 (13), 1028-1029 (1993),

Munoz, N. and Bosch, F. X., Epidemiology of cervical cancer. In: Human Papillomaviruses and Cervical Cancer, eds. Munoz, N., Bosch, F. X. and Jensen, O. M. IARC Scientific Publications, Lyon, 9-40 (1989)).

Broker et al., Papillomaviruses: Retrospectives and Prospectives, Cancer cells 4 / DNA tumor viruses, Cold Spraing Harbor Laboratory, USA, 17-36 (1989)

[7] Durst, M. et al., Papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographical regions, Proc. Natl. Acad. Sci., USA, 80. 3812-9-2 1019950024886, 3815 (1983)),

8, Hausen, H., Viruses in human cancers, Science, 254, 1173-1187 (1991)).

9 Galloway, D. A. et al., Human papillomaviruses and carcinomas, Adv. Virus Res., 37, 125-171 (1990)

10. Lorincz, A. T. et al., Human Papillomavirus Infection of the Cervix: Relative Risk Associations of 15 Common Anogenital Types, Obsterics and Gynecology, 79, 328-337 (1992)

[Reference 11] Kim, TJ, Korean Resources Plants . II , pp194-195, 1996

12. Kang, TH, Jeong, ST, et al., Journal of Ethnopharmacology, 71 , pp 321-323, 2000;

Reference Document 13 Jung, MJ, Chung, HY et al., Archives of pharmacal research , 26 (6) , pp458-462

Document 14 Kaneta M., Hikichi H. et al., Agricultural and Biological Chemistry , 44 , pp1407, 1980.

15. Woo WS, Kang SS et al., Journal of Natural Products , 49 , pp 547-549, 1984)

16. Woo WS, Lee EB et al., Kor . J. Pharmacogn , 12, pp 153, 1981

[Reference 17] Yang MS, Ha YL et al., Agricultural Chemistry and Biotechnology, 38 , pp584, 1995

[16] Yoo JS, Jang JS et al., KOR . J. Pharmacogn. , 26 , pp 355, 1995),

[Literature 18] Donald, BS et al., Phytochemistry , 27 (2), 1988, 517-522;

[Ref. 19] Sam-Sung-Sung, Shin Jung Hwa Medicine, pp 664-665, 1980

[Literature 20] Masayoshi Ogaga, Journal of the Japanese Pharmaceutical Sciences, p 336, 1991), Antonio, GG et al., Phytochemistry , 16 , 1977, 107-110

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[24] Norio, M. et al., Biosci . Biotechnol . Biochem . , 72 (2), 2008, 477-484

[Literature 25] Xian-Wen, Y. et al., Bioorg . Med . Chem ., 18 , 2010, 744-754;

[Literature 26] Lai-King, Sy. et al., J. Nat . Prod ., 61 , 1998, 907-912

[27] Thebtaranonth, CY et al. Phytochemistry , 40 (1), 1995, 125-128

Cancer is one of the incurable diseases that humanity has to solve, and huge capital has been invested in the development to cure it all over the world.In Korea, it is the number one disease cause of death among Koreans since 1983. More than 100,000 have been diagnosed, and more than 60,000 have died. Smoking, ultraviolet rays, chemicals, food, and other environmental factors are among the carcinogens that cause cancer, but the development of therapeutic agents is difficult due to the various causes, and the effects of the therapeutic agents are also different depending on the site where they occur.

Currently used anticancer agents include biological preparations such as enzyme preparations or vaccines, pure synthetic medicines, and medicines derived from natural products. Among them, anticancer drugs using genes, enzymes, vaccines, etc. are not in a practical stage and are used by chemotherapy. The developed anticancer drugs have considerable toxicity and have side effects of destroying not only cancer cells but also normal cells because they cannot selectively remove only cancer cells, and recently, cancer cells have become resistant to this and are ineffective for treating cancer. . Therefore, there is an urgent need to develop an effective anticancer agent that is less toxic for preventing cancer and develops cancer cells.

Apoptosis is an important mechanism of action in which most anticancer agents exhibit the antiproliferative effect of cancer cells (Kalechman et al., Synergistic anti-tumoral effect of paclitaxel (Taxol) + AS101 in a murine model of B16 melanoma: association with ras) -dependent signal-trasduction pathways Int J. Cancer , 86, pp.281-288, 2000;.. Gamet-Payractre et al ., Sulforaphane a naturally occurring isothiocyanate, induced arrest and apoptosis in HT29 human colon cancer cell. Cancer Res . , 60 , pp.1426-1433, 2000), which refers to active cell death regulated by genes and plays an important role in maintaining a balance between cell proliferation and cell death in vivo (Piao et al. al ., Induction of G (2) / M phase arrest and apoptosis by a new synthetic anti-cancer agent, DW2282, in promyelocytic leukemia (HL-60) cells. Bio chem Pharmacol , 62 , pp. 1439-1447, 2001).

Cervical cancer is a malignant tumor-causing virus that accounts for more than 90% of uterine cancers, and it is the most common cancer disease among Korean women. In recent years, infection with human papillomavirus (HPV) is the most important in carcinogenesis. It turns out to be a factor that acts.

Uterine cancer is currently the highest incidence among women in developing countries (Vanchieri, C., IARC Publishes Data on Worldwide Cancer Cases, Journal of the National Cancer Institute, 85 (13), 1028-1029 (1993)). It has been reported that there are about 500,000 patients (Munoz, N. and Bosch, FX, Epidemiology of cervical cancer.In: Human Papillomaviruses and Cervical Cancer, eds. Munoz, N., Bosch, FX and Jensen) , OM IARC Scientific Publications, Lyon, 9-40 (1989). According to a two-year study of cancer incidence from Seoul National University Medical School (Kim Jin-bok and Ahn Yun-ok, 1993) from July 1991 to June 1993 in Korea, 123 women per 100,000 people get cancer annually. The highest incidence of uterine cancer in Korea was 22%, stomach cancer 18%, and breast cancer 12.9%.

Papillomaviruses have been known to infect epithelial cells of various tissues of animals and cause benign tumors, commonly called warts, on the hands, feet, skin and larynx. These papillomaviruses are currently known to have about 70 genotypes in humans, and several genotypes have been reported to be specific for infectious tissues and cause various diseases (Broker et al., Papillomaviruses: Retrospectives and Prospectives, Cancer cells 4 / DNA tumor viruses, Cold Spraing Harbor Laboratory, USA, 17-36 (1989).

Most of these different genotype papillomaviruses were not noted because they were poorly treated and caused great pain in infected patients but did not cause fatal diseases. Recently, however, certain types of the virus, particularly human papillomavirus types 16 and 18, are associated with malignant tumors in the genitals, mouth, and skin of men and women and may act as a major factor in cervical cancer, which accounts for more than 90% of female uterine cancers. In addition, 6b and 11 have been shown to cause benign tumors of the male and female genitalia, called condyloma acuminata. Epidemiological studies have also suggested that uterine cancer is caused primarily by factors that are transmitted by sexual contact (Durst, M. et al., Papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographical regions, Proc). Natl.Acad.Sci., USA, 80. 3812-9-2 1019950024886, 3815 (1983)), papilloma in 85-100% of the prostate cancer lesions named cervical intraepithelial neoplasm (CIN). Because of the large body of evidence that papillomaviruses are involved in the development of uterine cancers, including the presence of viruses (Hausen, H., Viruses in human cancers, Science, 254, 1173-1187 (1991)), papillomaviruses In addition, studies on the carcinogenic mechanisms of cancer cells have been required as well as the development of diagnostic and therapeutic agents (Galloway, DA et al., Human papillomaviruses and carcinomas, Adv. Virus Res., 37, 125-171 (1990)).

On the other hand, the infection rate of human papillomavirus type 16 and 18 in uterine cancer patients is generally reported that type 16 to 50 to 70% and type 18 to 15 to 25%. However, in metastatic cancer, 25% of cancer patients infected with type 16 and more than 50% of cancer patients infected with type 18 were metastatic cancers (Lorincz, AT et al., Human Papillomavirus Infection of the Cervix: Relative Risk Associations of 15 Common Anogenital Types, Obsterics and Gynecology, 79, 328-337 (1992).

Among the plants native to Korea, pine needles are widely distributed in Korea, and have been used as folk medicines for insomnia, tonic, excitement, diuresis, pain relief, and insect repellent since ancient times (Kim, TJ, Korean). Resources Plants . II , pp 194-195, 1996).

Recently, many chemical components have been isolated from pine needles and their structure has been revealed (Kang, TH, Jeong, ST, et al., Journal of Ethnopharmacology , 71 , pp 321-323, 2000; Jung, MJ, Chung, HY et al., Archives of pharmacal research , 26 (6) , pp 458-462). For example, the study of pine needle components showed that quercetin 3-O-galactoside, a flavonoid, in the skin, quercetin-3-O-lamnoside, rhamnoside, 3,4,7-trihydroxyflavone and alpha-spinastery glucoside, several triterpenoids, saponin and lignan components And the like (Kaneta M., Hikichi H. et al., Agricultural and Biological Chemistry , 44 , pp 1407, 1980; Woo WS, Kang SS et al., Journal of Natural Products , 49 , pp547-549, 1984), and flavonol glycosides, quercitrin and isoquercitrin, were also isolated from flowers. In addition, studies on the pharmacological action of pine needle components showed that the saponin fraction increased the uterine contractility (Woo WS, Lee EB et al., Kor J. Pharmacogn, 12, pp 153, 1981) Ha YL et al., Agricultural Chemistry and Biotechnology , 38 , pp584, 1995), diarrhea inhibitory effect (Yoo JS, Jang JS et al., KOR . J. Pharmacogn , 26 , pp355, 1995), and sleep effect by flavonol glycoside. .

There are about 80-90 species of pines in the world, among them Pinus. Among them, P. palustris Miller (North America), P. pinaster Aiton (France), Pinus P. longifolia Rocvurgh (India), Pinus densiflora cepab (P. sylvestris L.), P. laricid poiret (Austria), P. longifolia Rocvurgh densiflora Sieb. et Zucc.: Korea, Japan), and P. thunberii Palatore (Haesong, Japan).

The main components of pine needles are Terpentine oil, Cineole, Salinigrin, Coniferin, P-Cymen, Densipimaric acid, Retten (Retene) and chlorophyll, protein, no fat, phosphorus, iron, enzymes, minerals, fat-soluble vitamin A and vitamin C, etc. The main ingredients are somewhat different depending on the species and the season harvested.

Essential oils obtained from the leaves of Pinus densiflora Sieb. Et Zucc. Include alpha-Pinene, beta-Pinene, camphene and pelanden ( phellandrene, borneol, bornyllacetate, caryophyllene, cadinene diterpene, sesquiterpene, sesquiterpenalcohol, cerylalcohol ), Beeswax contains juniperic acid, sabinic acid, hexadecane-diol, triacontan-1-ol, etc., and other machusterin ), Phytosterin, chinic acid, shikimic acid, quercetine, and kaempferol. (赤松 金 芳 ers, 신 정화 한 제, pp664- 665, 1980).

Among pines growing in Korea, pine needles are used as a medicine for gonorrhea and syphilis in folk medicine (御 影 雅 幸 雅, 日本 生 藥學 雜誌, p 336, 1991), but the pharmacological effects of pine needles are not known. And it is known to have the effect of restoring the coagulated skin to the original to the bruises of the skin quickly, and to heal eczema, scabies and sweat bands (Park Jong-gap, Korean Medicine, p134, 1984; Culture Broadcasting, Korean Civil Therapy Exhibition, p21, 1988). It is also mentioned in the appendix that it has the effect of preventing the whitening of hair. The herb wood is said to strengthen swelling, hair improvement, intestines and prolong life. Deodorant effect, insomnia healing effect, and skin beauty effect.

None of the above documents teaches or discloses that compounds isolated from pine needles have therapeutic effects on various cancer diseases.

Accordingly, the present inventors have confirmed the cytotoxicity against the compounds isolated from the pine needle leaf extract to cancer cells such as lung cancer, ovarian cancer, skin cancer, colon cancer, cervical cancer, etc., and found strong anticancer activity.

According to the above object, the present invention provides a pharmaceutical composition comprising ent-18-hydorxy-13-epimanoyl oxide (1), represented by the following Chemical Formula 1, Dehydroabietic acid 2, sandaracopimaric acid 3, 15-hydroxydehydroabietic acid 4, or caryophyllene oxide 5, Or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention also provides a pharmaceutical composition for prevention and treatment of cancer diseases.

Extracts as defined herein include crude extracts, fraction extracts or purified products.

The crude extract as defined herein is soluble in lower alcohols such as methanol, ethanol and butanol or a mixed solvent thereof, preferably a mixed solvent of 50-70% water and C1-C4 lower alcohol, more preferably 60% ethanol. One extract.

Fractional extracts as defined herein include nonpolar solvent soluble fraction extracts and polar solvent soluble fraction extracts.

The nonpolar solvent soluble fraction extract as defined herein is prepared by suspending the crude extract in water followed by treatment with a nonpolar solvent selected from hexane, methylene chloride, dichloromethane, chloroform or ethyl acetate, preferably hexane, methylene chloride fraction or ethyl acetate, More preferably a fraction extract which is soluble in a non-polar solvent obtained by fractionation with a methylene chloride nonpolar organic solvent; The polar solvent soluble fraction extract includes the fraction extract soluble in the polar solvent obtained by removing the non-polar solvent soluble tablet and fractionating the remaining crude extract with butanol or water.

The purified product as defined herein is about 1 to 30 times (w / w), preferably about 5 to 15 times (w / w), more preferably about 8 to 12 times (w / w) to the crude extract. SP207, HP20SS, Diaion HP 20, SP-850 resin, used for separating and purification of a water-soluble substance having the same weight as the weight of the purified water Activated carbon, Amberlite XAD-2,4, preferably Diaion HP 20, SP-850 resin, Amberlite XAD-2,4, and the mobile phase was eluted sequentially with ethanol, acetone, or methylene chloride solvent to lower the polarity, followed by elution (Hereinafter referred to as S11-HPO), 30% ethanol eluted tablets (hereinafter referred to as S11-HP30), 50% ethanol eluted tablets (hereinafter referred to as S11-HP50), 70% ethanol elution Purified water (hereinafter referred to as S11-HP70), 95% ethanol-eluted purified product (hereinafter referred to as S11-HP95), acetone and methylene chloride eluting purified product (hereinafter referred to as S11-HPAM).

Pine needles, as defined herein, are selected from the group consisting of Pinus densiflora Sieb. Et Zucc., Pinus rigida, Pinus taeda, Pinus thunbergii parlatore or Pinus koraiensis Sieb. Et Zucc The leaves of Pinus, preferably Pinus densiflora Sieb. Et Zucc., Pinus rigida, or Pinus taeda.

The cancer diseases defined herein include cancer of the lung, ovarian cancer, skin cancer, colon cancer, stomach cancer, breast cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, Endometrioid carcinoma, thyroid cancer, adenocarcinoma, small intestine cancer, small intestine cancer, endometrioid carcinoma, endometrioid carcinoma, carcinoma of the uterine cervix, vulvar carcinoma, vulvar carcinoma, Hodgkin's disease, Renal pelvic carcinoma, renal pelvic carcinoma (CNS), central nervous system (CNS), renal cell carcinoma, renal cell carcinoma, renal pelvic carcinoma, renal pelvic carcinoma, pancreatic cancer, Tumor, primary CNS lymphoma, spinal cord tumor, brain tumor, brain stem glioma, or pituitary adenoma, preferably lung cancer, ovarian cancer, skin cancer, colon cancer.

Extracts as defined herein include crude extracts or nonpolar solvent soluble extracts and may be prepared by the following process.

For example, the crude extract of the present invention may contain water, alcohol, and water containing 1 to 200 times (w / w), preferably 10 to 100 times (w / w) To 4% by weight of a lower alcohol or a mixed solvent thereof, preferably water or a mixed solvent of water and ethanol, more preferably water or a mixed solvent of 50% -99% water and ethanol, for 12 hours to 1 week, preferably 48 hours The extraction method such as cold extraction, hot water extraction, ultrasonic extraction and reflux cooling extraction at a temperature of from 10 to 150 ° C, preferably from 20 to 100 ° C, more preferably at room temperature, preferably, And then conducting a hydrothermal extraction method to obtain an extract.

For example, the non-polar solvent and polar solvent-soluble extract of the present invention may be used in an amount of about 0.0005 to 0.005 times, preferably 0.05 to 0.5 times (v / v) of the crude extract, preferably 60 to 90% w%) of water, followed by a conventional fractionation process using n-hexane, methylene chloride, ethyl acetate and butanol to obtain nonpolar solvent-soluble extraction tablets (n-hexane, water; And polar solvent soluble extract purified products which are soluble in polar solvents such as butanol and water, respectively.

For example, the purified product of the present invention is about 1 to 30 times (w / w), preferably about 5 to 15 times (w / w), more preferably about 8 to 12 times (w) to the crude extract. / w) purified water and SP207, HP20SS, Diaion HP 20, SP-850 resin, which is used for separation and purification of water-soluble substances having the same weight as the purified water. Activated carbon, Amberlite XAD-2,4, preferably Diaion HP 20, SP-850 resin, Amberlite XAD-2,4, and the mobile phase is sequentially eluted with a polarity lowering solvent such as ethanol, acetone or methylene chloride, followed by eluting with a water eluant (Hereinafter referred to as S11-HPO), a 30% ethanol eluate (hereinafter referred to as S11-HP30), a 50% ethanol elution solution (hereinafter referred to as S11- HP50), a 70% ethanol elution solution 95% ethanol eluate (hereinafter referred to as S11-HP95), acetone and methylene chloride elution solution (hereinafter referred to as S11-HPAM).

Accordingly, the present invention provides water, spirits, lower alcohols having 1 to 4 carbon atoms, including purified water of about 1 to 200 times (w / w), preferably 10 to 100 times (w / w) of the total weight of pine needle samples. Or a mixed solvent thereof, preferably water or water and ethanol mixed solvent, more preferably water or 50-99% water and ethanol mixed solvent, for 12 hours to 1 week, preferably 48 hours to 72 hours, 10 ℃ to 150 ℃, preferably 20 ℃ to 100 ℃, more preferably at room temperature extraction methods such as cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, preferably cold extraction or hot water extraction A first step of obtaining an extract; About 1 to 30 times (w / w), preferably about 5 to 15 times (w / w), more preferably about 8 to 12 times (w / w) of purified water are added to the crude extract. SP207, HP20SS, Diaion HP 20, SP-850 resin, used for the separation and purification of water-soluble substances of the same weight Activated carbon, Amberlite XAD-2,4, preferably Diaion HP 20, SP-850 resin, Amberlite XAD-2,4, and the second step of eluting the mobile phase with a polarity lowering solvent such as ethanol, acetone or methylene chloride solvent by sequentially performing an additional purification process using an adsorption chromatograph method using an adsorption resin such as Amberlite XAD- (Hereinafter referred to as S11-HPO), 30% ethanol eluted tablets (hereinafter referred to as S11-HP30), 50% ethanol eluted tablets (hereinafter referred to as S11-HP50) (Hereinafter referred to as S11-HP70), 95% ethanol-eluted tablets (hereinafter referred to as S11-HP95), acetone and methylene chloride-eluted tablets (hereinafter referred to as S11-HPAM) And a manufacturing method thereof.

In addition, conventional fractionation processes can also be carried out (Harborne JB, Phytochemical methods: A guide). to modern techniques of plant analysis , 3 rd Ed . , pp 6-7, 1998).

The compound of the present invention can be isolated from a plant extract such as a pine needle leaf extract or a chemical synthesis method common in the art. For example, the pine needles nonpolar solvent extract can be separated by silica gel chromatography using hexane: methylene chloride (1: 1), methylene chloride, hexane: methylene chloride: methanol (10: 10: 5, 10: 10: 2) and methylene chloride: methanol (1: The residue was separated by silica gel chromatography and eluted with hexane: ethyl acetate (20: 1, 10: 1, 5: 1, 3: 1), methylene chloride: methanol (3: 1) and 100% methanol, (46.11 g) was separated by silica gel chromatography to obtain compound 1, which was dissolved in hexane: ethyl acetate (7: 1 to 1: 1), hexane: ethyl acetate: methanol (10: 10: 5) and methylene chloride: methanol 1: 1) (A) and 12 fractions (B) were again chromatographed. Fractions 6 and 7 (A) were again separated by silica gel chromatography, and hexane (Eluent: 80% methanol), and the fractions 11, 12 and 13 of the obtained fractions were separated by chromatography on a LiChroprep RP-18 (40-63 μm, Merck, USA) (Fraction B) was subjected to chromatography using Sephadex LH-20 (eluent: methylene chloride: methanol = 4: 6) and LiChroprep RP-18 (40-63 [mu] m, Merck, USA) chromatography (eluent: 80% methanol).

The compounds of the present invention may be prepared into pharmaceutically acceptable salts and solvates by methods conventional in the art.

As the pharmaceutically acceptable salts defined herein, acid addition salts formed by free acid are useful. The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an acid aqueous solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and 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.

As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acids include methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid Citric acid, lactic acid, glycollic acid, gluconic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid and hydroiodic acid can be used.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving a compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the non-soluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt in particular, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the compounds of the present invention include, unless otherwise indicated, salts of acidic or basic groups that may be present in the compounds of the present invention. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of hydroxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate rate, methane sulfonate (mesylate) and p - toluene sulfonate (tosylate) and a salt, the salt manufacturing method or manufacturing process known in the art ≪ / RTI >

Compounds isolated from the pine needle extract of the present invention obtained by the above-described method have cytotoxicity against cancer cell lines such as lung cancer, ovarian cancer, skin cancer, colon cancer and cervical cancer. As a result, And can be used as a composition for treatment.

In addition, pine needles have been used as a herbal medicine for a long time, the extract of the present invention and compounds isolated therefrom also have no problems such as toxicity and side effects.

Pharmaceutical compositions for the treatment and prevention of cancer diseases of the present invention, the total weight of the composition of 0.02 to 50% It is included by weight percentage.

The pharmaceutical compositions comprising the compounds of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical compositions comprising the compounds according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, water, and the like in the extracts and tablets. Prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The preferred dosage of the compound of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or Intracerebroventricular injection.

In addition, the present invention relates to the use of ent-18-hydorxy-13-epimanoyl oxide (1), dehydroabietic acid a compound selected from dehydroabietic acid, 2, sandaracopimaric acid, 15-hydroxydehydroabietic acid, 4, or caryophyllene oxide, Or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention also provides a health functional food for preventing or ameliorating cancer diseases.

&Quot; Health functional food "as defined herein means food prepared and processed using raw materials or ingredients having functionality useful to the human body in accordance with Law No. 6727 on Health Functional Foods." Functional " Structure and function of the nutrient to control or physiological effects, such as to obtain a beneficial effect for health is intended to eat.

The health functional food for cancer prevention and improvement of the present invention contains 0.01 to 95% by weight, preferably 1 to 80% by weight, of the compound based on the total weight of the composition.

In addition, it is possible to manufacture and process as a health functional food in the form of tablets, capsules, powders, granules, liquids, pills for the purpose of improving and preventing cancer diseases.

For example, the health functional food in the form of tablets may be prepared as it is or granularly mixed with an excipient, a binder, a disintegrating agent or other additives in a suitable manner, and then compressed into a glidant, etc. It is made by directly compressing the functional food as it is or by mixing it evenly with an excipient, binder, disintegrant or other suitable additives, or mixing the health functional food as it is or evenly adding the appropriate additive to the prepared granules, The excipient, binder or other suitable additives are added to the functional food, and the powder mixed evenly is wetted with a solvent, the wet powder is molded into a mold at low pressure, and then dried and prepared by a suitable method. In addition, a mating agent or the like may be added to the health functional food in the form of tablets, if necessary.

Among the health functional foods in the form of capsules, the hard capsules are usually prepared by mixing the capsules evenly with the health functional foods or excipients suitable for the health functional foods, granulated by a suitable method, or granulated with a suitable epidermal agent as it is or Soft capsules are prepared by filling them, and soft capsules are usually filled with capsules containing glycerin or sorbitol in a capsule form containing gelatin or appropriate excipients suitable for health functional foods or health functional foods. It is molded and prepared, and a coloring agent preservative etc. can be added to the said capsule base as needed.

Circular functional foods are usually mixed with excipients, binders, disintegrants, etc., and then molded into a spherical form in a suitable manner, and the coating is carried out with white sugar or other suitable coating agent, or starch, talc or You can also be greeted with a suitable substance.

The health functional food in the form of granules is usually made by mixing the health functional food as it is or by adding excipients, binders, disintegrating agents, etc., evenly and then granulating it in a proper way to make the particles as even as possible. , Mating agent, etc. can be added. For the health functional food in the form of granules, No. 12 (1680 μm), No. 14 (1410 μm) and No. 45 (350 μm) are used for the next particle size test. It should be less than 5.0% and pass through No.45 to less than 15.0% of the total amount.

The definitions of the excipients, binders, disintegrants, lubricants, mating agents, flavoring agents, etc. of the present invention are described in documents known in the art and include the same functions or the like (see, for example, , Council of Korean Pharmacy College, 5th ed., P. 33-48, 1989).

The compound isolated from the pine leaf extract of the present invention shows cell growth inhibitory effect in various cancer cells and is useful for pharmaceutical compositions and health functional foods for the prevention and treatment of various cancer diseases including lung cancer, ovarian cancer, skin cancer, colon cancer, cervical cancer and the like Can be used.

Hereinafter, the present invention will be described in detail with reference to the following examples and experimental examples.

However, the following examples and experimental examples are illustrative of the present invention, and the content of the present invention is not limited by the following examples and experimental examples.

Example 1 Preparation of Ethanol Extract of Pine Needle

2 kg of Gangwon-do pine needles purchased from Gyeongdong market were extracted twice with 50 L of 95% ethanol (alcohol) at 50 ° C., and concentrated under reduced pressure to obtain 338.85 g of ethanol extract.

Example  2. Pine needles Fractions  Produce

The ethanol extract of Example 1 was suspended in 1.8 L of distilled water and the fractions were sequentially fractionated three times with the same amount of hexane, methylene chloride, ethyl acetate and n-butanol in accordance with the conventional method to obtain a hexane fraction (95.18 g) and a methylene chloride fraction ), Ethyl acetate fraction (19.42 g), n-butanol fraction (57.94 g) and water fraction (90.2 g).

Example  3. Ethanol extract of pine needles HP -20 Column  Used Small fraction  Produce

30 g of pine needle ethanol extract obtained in Example 1 was suspended in 30 ml of water, followed by Diaion HP-20 column chromatography. First, 2 L of water was added to the mobile phase to obtain an eluate, and 2 L of 30% ethanol, 2 L of 50% ethanol, 2 L of 70% ethanol, 2 L of 95% ethanol, 2 L of acetone and 1.5 L of methylene chloride were added respectively. HP70 (water, 7.95g), HP30 (30% ethanol, 3.44g), HP50 (50% ethanol, 2.97g) and HP70 (70%) were mixed with the acetone eluate and methylene chloride eluent, Ethanol, 1.75 g), HP95 (95% ethanol, 6.35 g) and HPAM (acetone and methylene chloride, 6 g).

Example  4. Pine needles From the fraction  Active substance separation

The hexane fraction (95.18 g) obtained in Example 2 was separated by silica gel chromatography and eluted with hexane: methylene chloride (1: 1), methylene chloride, hexane: methylene chloride: methanol (10: 10: 0.5, ) And a mixed solvent of methylene chloride: methanol (1: 1). Of these, 4.06 g of fractions 5 and 6 were separated again by silica gel chromatography. Separation was carried out with an eluent of hexane: ethyl acetate (20: 1, 10: 1, 5: 1, 3: 1), methylene chloride: methanol (3: 1) and 100% methanol and 58.8 mg of compound 1 was obtained. The methylene chloride fraction (46.11 g) was then separated by silica gel chromatography and eluted with hexane: ethyl acetate (7: 1-1: 1), hexane: ethyl acetate: methanol (10: 10: 0.5) and methylene chloride: methanol 1: 1). Of these, 405.9 mg of fraction 6 (A) and 864.7 mg of fraction 12 (B) were chromatographed again. Further, 407.9 mg of fraction 6 (A) was again separated by silica gel chromatography. Separation was carried out with an eluent of hexane: ethyl acetate (10: 1), and fractions 11, 12 and 13 (74.9 mg) were obtained. This fraction was separated by chromatography on LiChroprep RP-18 (40-63 μm, Merck, USA) (eluent: 80% methanol) to give 4 mg of compound 2, 32.1 mg of compound 3 and 2.9 mg of compound 4, respectively . 864.7 mg of fraction 12 (B) was separated by chromatography on Sephadex LH-20 (eluent: methylene chloride: methanol = 4: 6) to give 441 mg fractions. This fraction was subjected to LiChroprep RP-18 (40-63 mu m, Merck, USA) chromatography (eluent: 80% methanol) to give 106.3 mg of Compound 5. The chemical structures of compounds 1,2,3,4,5 could be confirmed by comparing the spectra with known data.

Compounds 1 to 5 could be identified from a comparison of the results cited below with the references cited below. These are as follows.

(1) ent-18-hydorxy-13-epimanoyl oxide (hereinafter referred to as compound 1) (Reference: Donald, BS et al., Phytochemistry , 27 ), 1988, 517-522; Antonio, GG et al., Phytochemistry , 16 , 1977, 107-110)

m.p. 47-48 ° C

_{[α] D +13.1 (CHCl 3} , c0.013)

ESI MS (positive) m / z 329 [M + Na] < + &

¹ H-NMR (CDCl ₃ , 500 MHz)? 0.72 (3H, s, H-20), 0.97 D, J = 10.5H, H-18b), 4.91 (1H, br d, J = 11.0H, (1H, d, J = 17.5, 11.0H, H-14), 4.96 (1H, br d, J = 17.5H, H-

¹³ C-NMR (CDCl ₃ , 125 MH)? 16.2 (C-11), 16.5 (C-20), 18.5 38.9 (C-3), 39.7 (C-1), 43.7 (C-12), 32.9 (C-15), 147.9 (C-14), 73.5 (C-13), 76.1 )

(2) Dehydroabietic acid (hereinafter referred to as Compound 2) [Reference: Hyun Jin, J. et al., Arch . Pharm . Res . , 34 (6), 2011, 913-917; Miguel, AG et al., Eur. J. Med. Chem. , ≪ / RTI > 45 , 2010, 811-816)

_{[α] D +59.7 (CHCl 3} , c0.5)

ESI MS (positive) m / z 323 [M + Na] +

¹ H-NMR (CDCl ₃ , 500 MHz)? 1.21 (3H, s, H-20), 1.22 (6H, d J = 6.0H, H- (1H, br s, H-14), 6.88 (1H, d, J = 1.5H, H- 4.0H, H-12), 7.17 (1H, d, J = 8.0H, H-11)

_{^{13 C-NMR (CDCl 3,}} 125 MH) δ 16.5 (C-19), 18.8 (C-2), 20.0 (C-6), 24.2 (C-16,17), 25.3 (C-20), 30.2 (C-7), 33.7 (C-15), 37.0 (C-3), 37.1 (C-10), 38.2 C-12), 124.3 (C-11), 127.1 (C-14), 134.9 (C-8), 146.0

(3) sandaracopimaric acid (hereinafter referred to as Compound 3) [Reference: Nguyen Hai, D. et al., Arch . Pharm . Res . , 28 , 2005, 28-33; Norio, M. et al., Biosci. Biotechnol. Biochem. , 72 (2), 2008, 477-484)

m.p. 162-163 DEG C

[?] _D -5.2 (CHCl ₃ , c 0.43)

ESI MS (positive) m / z 324 [M + Na] < + &

¹ H-NMR (CDCl ₃ , 500 MHz)? 0.84 (3H, s, H-20), 1.04 D, J = 17.5, 1.5H, H-16a), 5.22 (1H, s, H-14), 5.77 , 11.0H, H-15)

_{^{13 C-NMR (CDCl 3,}} 125 MH) δ 15.4 (C-20), 17.0 (C-19), 18.4 (C-2), 18.8 (C-11), 25.1 (C-6), 26.2 (C 37.7 (C-13), 38.0 (C-10), 38.5 (C-1), 47.5 (C- (C-15), 185.5 (C-15), 49.0 (C-5), 50.8 (C-9), 110.4 )

(4) 15-Hydroxydehydroabietic acid (hereinafter referred to as Compound 4) [Reference: Xian-Wen, Y. et al., Bioorg . Med . Chem., 18 , 2010, 744-754; Lai-King, Sy. et al., J. Nat. Prod., 61 , 1998, 907-912)

_{[α] D +113.7 (CHCl 3} , c0.02)

ESI MS (positive) m / z 339 [M + Na] < + &

¹ H-NMR (CDCl ₃ , 500 MHz)? 1.20 (3H, s, H-20), 1.26 (3H, br s, H-19), 1.54 1H, br s, H-14), 7.20 (2H, m, H-11, 12)

¹³ C-NMR (CDCl ₃ , 125 MH)? 16.5 (C-19), 18.7 (C-2), 21.9 (C-16), 37.1 (C-16,17), 38.1 (C-3), 44.8 C-14), 124.4 (C-12), 125.1 (C-11), 135.0

(5) caryophyllene oxide (hereinafter referred to as Compound 5) [Reference: Thebtaranonth, CY et al., Phytochemistry , 40 (1), 1995, 125-128].

m.p. 60-62 ° C

_{[α] D -70.12 (CHCl 3} , c0.65)

ESI MS (positive) m / z 243 [M + Na] < + &

¹ H-NMR (CDCl ₃ , 500 MHz)? 0.99 (3H, s, H-12), 1.02 , H-15 [alpha]), 4.99 (1H, m, H-15 [beta]

_{^{13 C-NMR (CDCl 3,}} 125 MH) δ 21.7 (C-13), 27.2 (C-2), 29.8 (C-7), 30.0 (C-12), 30.3 (C-6), 34.0 (C (C-3), 39.8 (C-10), 48.8 (C-9), 50.7 (C-1), 63.7 8)

Experimental Example  1. Cytotoxicity test on human tumor cell line

In order to confirm the cytotoxicity of the above-mentioned samples to human tumor cell lines, an experiment was conducted as follows (see Skehan, P. et al., J. National Cancer Institute, 82: 1107 1990).

The six fractions obtained in the above example showed activity (cytotoxicity) of cervical cancer and cervical cancer of human uterine carcinoma (MES-SA) and multidrug resistant carcinoma subline of MES-SA (MES-SA). Was observed [Table 1].

In addition, the activity of the human uterine carcinoma (MES-SA), MES-SA / DX5 (multidrug resistant carcinoma subline of MES-SA) against cervical cancer and resistant cervical cancer (cytotoxicity) It was observed that [Table 2].

SK-OV-3 (human ovarian tumor), SK-MEL-2 (human malignant melanoma), HCT15 (human colorectal adenocarcinoma) and A-549 (human lung adenocarcinoma) , MES-SA (human cervical cancer), and MES-SA / DX5 (resistant cervical cancer) to human tumor cell lines (Table 3).

For cytotoxicity, six human cancer cell lines [A-549 (human lung adenocarcinoma, NCI), SK-OV-3 (human ovarian cancer cell line, (Human cervical cancer, NCI), MES-SA / DX5 (human resistant cervical cancer, NCI), SK-MEL-2 (human malignant melanoma, NCI), HCT15 NCI) was used. Passaging cancer cells are detached from the instrumentation side with trypsin-EDTA solution, and then the number of cells per well in a 96-well flat bottom microplate is 5 × 10 ³ ( A-549, HCT15), 1 × 10 ⁴ (SK-MEL-2), and 5 × 10 ³ (SK-OV-3). These divided cancer cells were cultured in a CO ₂ incubator (MCO-20AIC, SANYO Electric Co., LTd.), Attached to the bottom, and then removed with media using an aspirator and diluted with 6 log concentrations of sample solution. Into the well containing 100 ㎕ 3 times and incubated for 48 hours. After 48 hours of incubation, the media of each well was removed, and 100 µl of 10% trichloroacetic acid was added thereto, and the cells were left at 4 ° C. for 1 hour to fix the cells on the bottom of the plate. After the cell was fixed, the plate was washed 5 to 6 times with distilled water to completely remove the remaining trichloroacetic acid solution and dried at room temperature. Completely dried plate was stained with 100% of 1% acetic acid solution per well in 0.4% SRB (sulforhodamine B, Sigma) solution and stained for 30 minutes, then 1% acetic acid. 5-6 times to remove excess SRB not bound to cells. The stained cell plates were dried again at room temperature, and then 100 μl of 10 mM trisma base solution per well was added and shaken with a titer plate shaker (KOMA Orbital Shaker KE011, KOMABIOTech) for 10 minutes. After elution, the absorbance was measured at 520 nm using a microplate spectrophotometer (Sunrise, TECAN).

Cytotoxicity of ethanol extract and pine needle fraction, hexane fraction, methylene chloride fraction, ethyl acetate fraction, n-butanol fraction, water fraction and doxorubicin to cervical cancer and resistant cervical cancer Cell line IC ₅₀ (μg / ml) Ethanol extract Hexane fraction Methylene chloride fraction Ethyl acetate fraction n-butanol fraction Water fraction doxorubicin MES-SA 22.69 11.04 11.73 112.47 257.39 > 300.0 0.0002 MES-SA / DX5 23.14 16.28 12.92 186.94 > 300.0 > 300.0 0.4122 ratio* 1.02 1.47 1.10 1.66 - - 2061 * Ratio is the numerical value of MES-SA / DX5 divided by the numerical value of MES-SA.

Cytotoxicity of Small Fractions HP0, HP30, HP50, HP70, HP95, HPAM and Doxorubicin Against Cervical and Resistant Cervical Cancer Cell line IC ₅₀ (μg / ml) HP0 HP30 HP50 HP70 HP95 HPAM doxorubicin MES-SA > 300.0 > 300.0 188.63 56.67 13.44 15.83 0.0002 MES-SA / DX5 > 300.0 > 300.0 286.32 81.35 11.94 16.70 0.4122 ratio* - - 1.52 1.44 0.89 1.05 2061 * Ratio is the numerical value of MES-SA / DX5 divided by the numerical value of MES-SA.

The cytotoxicity of compounds 1-5 and doxorubicin on human tumor cell lines compound IC ₅₀ (μg / ml) A-549 SK-OV-3 SK-MEL-2 HCT15 MES-SA One 13.34 12.70 11.26 10.04 8.17 2 13.44 17.29 13.02 15.68 10.33 3 13.64 12.14 11.17 13.58 10.45 4 109.07 141.77 88.82 98.25 90.66 5 36.36 23.71 18.64 26.75 15.25 Doxorubicin 0.0072 0.0104 0.0012 0.0833 0.0051

As a result of the above experiments, the compounds of the present invention 1 to 5 were found to be effective against A-549 (human lung adenocarcinoma), SK-OV-3 (human ovarian tumor), SK-MEL-2 (human malignant melanoma), HCT15 , MES-SA (human cervical cancer) and MES-SA / DX5 (human resistant cervical cancer) human tumor cell lines.

Hereinafter, an example of the preparation of a pharmaceutical composition containing the compound of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Formulation example  One. Sanje  Produce

Compound 1 300 mg

Lactose 100 mg

Talc 10 mg

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

Formulation example  2. Preparation of tablets

Compound 2 300 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.

Formulation example  3. Preparation of capsules

Compound 3 300 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

Compound 4 300 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation example  5. Liquid  Produce

Compound 5 300 mg

10 g per isomer

5 g mannitol

Purified water

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, 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 adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Manufacture of health food

Compound 1 1000 mg

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg of vitamin B ₁

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

10 mg vitamin C

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

24.8 mg of magnesium chloride

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.

Formulation example  7. Manufacture of health drinks

Compound 1 1000 mg

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

After mixing the above components in accordance with the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed, sterilized and refrigerated It is used to prepare a healthy beverage composition of the present invention. Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (6)

Ent-18-hydorxy-13-epimanoyl oxide (1), dehydroabietic acid, 2, isolated from pine needle extract ), Sandaracopimaric acid (3), 15-hydroxydehydroabietic acid (15), or a compound selected from caryophyllene oxide (caryophyllene oxide 5) or a pharmaceutically thereof A pharmaceutical composition for preventing and treating cancer diseases containing an acceptable salt as an active ingredient.
≪ Formula 1 >

The method of claim 1,
The pine leaves include leaves of Pinus densiflora Sieb. Et Zucc., Pinus rigida, Pinus taeda, Pinus thunbergii parlatore or Pinus koraiensis Sieb. Et Zucc. ≪ / RTI > According to claim 1, wherein the cancer disease is lung cancer, ovarian cancer, skin cancer, colon cancer, stomach cancer, breast cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, head or neck cancer, skin or eye melanoma, uterine cancer, ovarian cancer, colon cancer , Small bowel cancer, rectal cancer, anal muscle cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, lymph gland cancer, bladder cancer, gallbladder cancer, endocrine cancer , Thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocyte lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS; central) nervous system) A pharmaceutical composition that is one or more diseases selected from the group consisting of tumors, primary CNS lymphomas, spinal cord tumors, brain tumors, brain stem glioma or pituitary adenoma. The pharmaceutical composition of claim 1, wherein the cancer disease is lung cancer, ovarian cancer, skin cancer, or colon cancer. Ent-18-hydorxy-13-epimanoyl oxide (1), dehydroabietic acid, 2, isolated from pine needle extract ), Sandaracopimaric acid (3), 15-hydroxydehydroabietic acid (15), or a compound selected from caryophyllene oxide (caryophyllene oxide 5) or a pharmaceutically thereof A health functional food for preventing and improving cancer diseases containing an acceptable salt as an active ingredient.
≪ Formula 1 >

The health functional food according to claim 5, wherein the health functional food form is powder, granule, tablet, capsule or beverage.