KR20050047207A - Composition containing an extract of zingiber cassumunar or the compound isolated therefrom for preventing and treating cancer disease - Google Patents

Abstract

The present invention relates to a novel anticancer agent isolated from a natural substance. Specifically, a composition comprising a ginger ( Zingiber cassumunar Roxb.) Extract of the present invention or a compound isolated therefrom exhibits cytotoxicity against various human cancer cells. It can be used as a medicine and health functional food for the prevention and treatment of cancer diseases.

Description

Composition containing an extract of Zingiber cassumunar or the compound isolated therefrom for preventing and treating cancer disease}

The present invention relates to a composition for the prevention and treatment of cancer diseases comprising a ginger extract or a compound isolated therefrom.

Cancer is one of the incurable diseases that humanity has to solve, and huge capital is invested in the development to cure it all over the world.In Korea, it is the number one disease death cause and more than 100,000 people a year It is diagnosed, and about 60,000 or more die. Carcinogens, which cause cancer, are smoking, ultraviolet rays, chemicals, foods, and other environmental factors. However, various causes are not only difficult to develop a therapeutic agent, but also effective effects vary depending on the site of occurrence. Currently, the substances used as therapeutic agents are extremely toxic and cannot selectively remove only cancer cells. Therefore, there is an urgent need for the development of low-toxic and effective anti-cancer agents to prevent cancer after treatment. .

Cancer is characterized by "uncontrolled cell growth," which results in the formation of cell masses called tumors that infiltrate surrounding tissues and, in severe cases, metastasize to other organs of the body. Academia is also called neoplasia. Cancer is an intractable chronic disease that, even if treated with surgery, radiation, and chemotherapy, in many cases does not heal fundamentally, suffers the patient, and ultimately leads to death. There are many factors that cause cancer, but they can be divided into internal and external factors. It is not known exactly how the normal cells are transformed into cancer cells, but at least 80 to 90% is known to be affected by external factors such as environmental factors. Internal factors include genetic factors, immunological factors, and external factors include chemicals, radiation, and viruses. Genes involved in the development of cancer include oncogenes and tumor suppressor genes, which occur when the balance between them is broken down by internal or external factors described above.

Cancer is classified into blood cancer and solid cancer, and it occurs in almost every part of the body such as lung cancer, stomach cancer, breast cancer, oral cancer, liver cancer, uterine cancer, esophageal cancer, and skin cancer. Among the methods used to treat these malignancies, chemotherapy agents, except surgery or radiation therapy, are collectively called anticancer agents, and most of them show anticancer activity by inhibiting the synthesis of nucleic acids. Chemotherapeutic agents are broadly classified into antimetabolites, alkylating agents, antimitotic drugs, and hormones, and folates that inhibit metabolic processes necessary for the proliferation of cancer cells. Derivatives (methotrexate), purine derivatives (6-mercaptopurine, 6-thioguanine), pyrimidine derivatives (5-fluorouracil, Cytarabine), etc.Introducing alkyl groups to guanine of DNA to modify the structure of DNA and chain-cutting Alkylating agents exhibiting effects include nitrogen mustard compounds (chlorambucil, cyclophosphamide), ethyleneimine compounds (thiotepa), alkylsulfonate compounds (busulfan), nitrosourea compounds (carmustine), triazene compounds (dacarbazine) There is. Mitosis inhibitors that inhibit mitosis by blocking mitosis as a specific period of mitosis, include anticancer drugs such as actinomycin D, doxorubicin, bleomycin, mitomycin, and plants such as vincristine and vinblastine. Alkaloids, taxoids including mitosis inhibitors including taxane rings, and the like. In addition, hormones such as corticosteroids and progesterone and platinum-containing compounds such as cisplatin are used as anticancer agents.

The biggest problem with chemotherapeutic agents is drug resistance, which, despite the initial successful response by anticancer agents, is a major factor that eventually causes treatment to fail. In connection with the problem of overcoming these side effects, efforts have recently been made to find the active ingredient in natural products that are used in the private sector.

Ginger ( Zingiber cassumunar Roxb.) Of the present invention is a genus Zingiber (Zingiber) belonging to the plant (Zingiberaceae), widely distributed in Southeast Asia, and has been used as a folk remedy. Examples of the use of creeper plants as a folk remedy include the fact that the roots of various creeper plants in Malaysia include stomach pain, motion sickness, vomiting, epilepsy, sore throat, cough, bruises, wounds, dissolution, eye disease, liver disease, rheumatism, myalgia, ringworm ( Ringworm), asthma, fever, malignancy and boils. In addition, the components of the genus Citrus plants are curcuminoids (Ruby et al., Cancer Lett., 94 , pp79-83, 1995; Ishida et al., Bioorg. Med. Chem., 10 , pp3481-3487, 2002; Vimala et al., Br. J. Cancer, 80 , pp110-116, 1999), flavonoid glycosides (Nakatani et al., Agric. Biol. Chem. , 55 , pp455-460, 1991; Murakami et al., Phytochemistry, 31 , pp2689-2693, 1992), sesquiterpenoids (Murakami et al., Carcino genesis, 23 , pp795-802, 2002) and Polyphenols (Murakami et al., Phytochemistry, 31 , pp2689-2693, 1992) and the like have been reported.

Ginger root has been used in Indonesia as a traditional medicine for treating abdominal pain, diarrhea, hookworm, pain, rheumatism and uterine disease (Ozaki et al., Chem. Pharm. Bull., 39 , pp2353-2356, 1991). In addition, studies on activity include antioxidant activity (Masuda et al., Chem. Lett., 1 , pp 189-192, 1993; Jitoe et al., Tetrahedron Lett., 35 , pp981-984, 1994), anti-inflammatory action (Ozaki et al., Chem. Pharm. Bull., 39 , pp2353-2356, 1991; Jeenapongsa et al., J. Ethnopharmacol. , 87 , pp143-148, 2003), insecticidal action (Nugroho et al., Phytochemistry , 41 , pp129-132, 1996) and uterine relaxation (Kanjanapothi et al., Planta Med. , 53 , pp329-332, 1987) have been reported, and phenylbutenoid and curcume have been studied in the root component of ginger. Curcuminoids have been reported (Masuda and Jitoe, Phytochemistry, 39 , pp 459-461, 1995; Tuntiwachwuttikul et al., Phytochemistry, 20 , 1164-1165, 1981; Masuda et al., Chem. Lett., 1 , pp 189 -192, 1993; Jitoe et al., Tetrahedron Lett., 35 , pp981-984, 1994).

However, despite continuing research on isolated substances from ginger, no teaching or suggestion has been made of the separation of potent substances for cytotoxicity from this plant.

Accordingly, the present inventors have completed the present invention by confirming that the compounds of the present invention isolated from ginger have excellent anticancer effects as a result of studying cancer drug treatment agents of herbal origin, which are widely used in folk medicine.

An object of the present invention is to provide a pharmaceutical composition and health functional food for the prevention and treatment of cancer diseases comprising a ginger extract having an anticancer activity or a compound isolated therefrom as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing and treating cancer disease containing ginger ( Zingiber cassumunar Roxb.) Polar solvent soluble extract or non-polar solvent soluble extract as an active ingredient.

The polar solvent soluble extract includes water, a lower alcohol having a carbon number of C ₁ to C ₄ or a mixed solvent thereof, preferably an extract soluble in methanol, and the non-polar solvent soluble extract includes hexane, ethyl acetate, chloroform and the like. Extracts soluble in a nonpolar solvent, preferably chloroform.

The present invention also provides a pharmaceutical composition for the prevention and treatment of cancer diseases comprising the compound of formula (I) or a pharmacologically acceptable salt thereof isolated from ginger as an active ingredient.

(I)

Where

R ₁ , R ₂ , R ₃ and R ₄ are each independently -H, -OR ₅ , -SR ₅ or -NR ₅ ,

R ₅ is hydrogen, lower alkyl or alkenyl group having 1 to 5 carbon atoms.

Among the compounds of the general formula (I), preferred compounds are those in which R ₁ to R ₄ are -OR ₅ in the compound of the general formula (I), and more preferably (±) -trans-3 of the structural formula (Ia) -(4-hydroxy-3-methoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohexene-1 -ne and (±) -trans-3 of formula (Ib) -(3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohex-1-cene.

(Ia)

(Ib)

In addition, the compounds include Alpinia katsumadai Hayata, Alpinia officinarum Hance, Alpinia oxyphylla Miq, Amomum cardamomum Linn., Amomum villosum Lour., Curcuma aromatica Salisb .), Curcuma longa Linn., Curcuma zedoaria Rosc., Zingiber mioga (Thunb. Rosc.) And Ginger ( Zingiber cassumunar Roxb.) Plants, preferably obtained separately from ginger have.

The cancer disease includes a general cancer disease, preferably gastric cancer, colon cancer, breast cancer, lung cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or eye melanoma, uterine cancer, ovarian cancer, rectal cancer , Anal leiomyoma, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma Cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, renal or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, One or more diseases selected from brain stem glioma, pituitary adenoma, and the like.

Hereinafter, the present invention will be described in more detail.

Ginger ( Zingiber cassumunar Roxb.) Polar solvent extract and non-polar solvent extract of the present invention can be obtained as follows.

Ginger ( Zingiber cassumunar Roxb.) Of the present invention is dried, chopped roots, and then water of about 1 to 20 times, preferably about 3 to 10 times the weight of the sample (g), C ₁ to C ₄ Lower alcohols or a mixed solvent thereof, preferably methanol, at an extraction temperature of 20 ° C. to 100 ° C., preferably 50 ° C. to 70 ° C. for about 1 to 10 days, preferably about 2 to 5 hours. After recovering the supernatant using cold needle, hot water extraction, ultrasonic extraction, reflux cooling extraction, more preferably reflux cooling extraction method, repeat the above procedure 2 to 7 times to collect the supernatant and concentrated under reduced pressure to ginger ginger Solvent soluble extracts can be obtained.

In addition, the non-polar solvent soluble extract of the present invention, after suspending the polar solvent soluble extract in distilled water, the suspension is about 1 to 100 times, preferably about 1 to 5 times the volume of hexane, ethyl acetate, methylene chloride, chloroform By adding a non-polar solvent such as 1 to 10 times, preferably 2 to 5 times can be obtained by extracting and separating the non-polar solvent soluble layer. It is also possible to further carry out conventional fractionation processes (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis., 3rd Ed. , Pp 6-7, 1998).

More specifically, the fractions having high activity in the ginger non-polar solvent soluble extract obtained by the above process are taken, the fractions having similar R _f values are collected on TLC to form a single fraction, and dried to obtain a purified fraction of purified form. With methylene chloride, chloroform, hexane, ethyl acetate, methanol, distilled water or a mixed solvent thereof, silica gel column chromatography is preferably carried out using a methylene chloride: methanol mixed solvent to separate into 5 to 13 fractions. Can be. The separated fractions were fractionated again to obtain (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] of the above formula (Ia). Cyclohexen-1-cene and (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohex-1 of formula (lb) -Cene compounds can be obtained.

Compounds having a prophylactic and therapeutic effect of cancer diseases of the present invention can be synthesized through the synthesis and fractionation method of conventional substituents (Herbert O. House: Modern Synthetic Reactions , 2nd Ed ., The Benjamin / Cummings Publishing Co., 1972).

The compounds of the present invention represented by the general formula (I) may be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.

As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equimolar amounts of the compound and acid or alcohol (eg, glycol monomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

At this time, it is possible is to use an organic acid and an inorganic acid as the free acid, the number of inorganic acids are used, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. and organic acid is methanesulfonic acid, p - toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salt, 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 formula (I) include salts of acidic or basic groups which may be present in compounds of formula (I) unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.

Compound of the general formula (I) of the present invention obtained by the above method, preferably (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E ) -3, 4-dimethoxystyryl] cyclohex-1-cene and (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohex To investigate the anticancer effects of the 1-Sen compound, in vitro (A549 (human lung cancer cells), Col2 (human colon cancer cells), SNU-638 (human gastric cancer cells) and HT-1080 (human fibroid tumors) As a result of the cytotoxicity of the cells), it was confirmed that the compounds of the present invention exhibited an excellent cytotoxic effect on cancer cell lines.

The pharmaceutical composition for preventing and treating cancer diseases of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition.

Pharmaceutical compositions comprising the extract or compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the extracts or 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 composition comprising the extract or compound according to the present invention may be prepared according to a conventional method, respectively, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions. It can be formulated and used in the form. Carriers, excipients and diluents that may be included in the composition comprising the extract or compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are 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, sucrose, or the like in the extract or compound. It is 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. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts or compounds of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract or compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

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

The present invention provides a dietary supplement comprising the extract, a compound, and a food-acceptable food supplement additive that exhibits a prophylactic effect of a disease caused by cancer. Examples of the food to which the extract or compound of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to foods or beverages for the purpose of preventing the disease caused by cancer. At this time, the amount of the extract or compound in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml Can be added.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract or compound as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. have. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the extracts or compounds of the present invention are various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the extracts or compounds of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

The present invention is described in more detail based on the following examples and experimental examples, but the present invention is not limited thereto.

Example 1 Ginger Zingiber cassumunar  Roxb.) Preparation of Crude Extracts

Ginger ( Zingiber cassumunar Roxb.) Was collected and dried in Surabaya, Indonesia, and 500 g of the sample obtained by cutting the roots was placed in 10 l of 100% methanol, and heated in a water bath using a reflux condenser. Extraction for 3 hours. This process was repeated twice to collect the supernatant, and then concentrated under reduced pressure to obtain 50 g of crude ginger extract.

Example 2 Preparation of Ginger Nonpolar Solvent Fractions

50 g of the crude crude extract obtained in Example 1 was suspended in 1 L of distilled water, and then dissolved by adding 1 L of hexane, and then vacuum dried by separating only the components dissolved in the hexane layer. This process was repeated five times to give 14.3 g of hexane fraction. 1 L of chloroform was added to the remaining aqueous layer, and only the components dissolved in the chloroform layer were separated and dried in vacuo. This process was repeated five times to give 12.9 g of chloroform fraction. The same procedure as described above gave 3.3 g of n-butanol fraction and 11.4 g of aqueous layer.

Samples were prepared by serial dilutions of each fraction in 5-fold serial concentrations at a maximum concentration of 20 μg / ml and used for drug efficacy experiments.

Example 3. (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E Separation and Purification of 3) -3,4-dimethoxystyryl] cyclohexen-1

13.0 g of the chloroform fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (silica gel 60, 230 to 400 mesh, using methylene chloride: methanol mixed solvent (100: 1 → 10: 1)). Merck), and separated into 13 fractions, and 1.8 g of fraction 1 of these fractions was again subjected to silica gel column chromatography (silica gel 60, 230) using a hexane: ethyl acetate mixed solvent (100: 0 → 0: 100). To 400 mesh, Merck). 650 g of the combined fractions from Nos. 9 to 10 were again subjected to silica gel column chromatography (silica gel 60, 230 to 400 mesh, Merck) using a hexane: acetone mixed solvent (70: 1 → 0: 100). And fraction (6) derived therefrom from the active ingredient (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl ] 11 mg of cyclohexen-1-cene (named Compound 1) was isolated.

Example 4. (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E Separation and Purification of) -3,4-dimethoxystyryl] cyclohexen-1

13.0 g of the chloroform fraction having the highest activity among the solvent fractions obtained in Example 2 was subjected to silica gel column chromatography (silica gel 60, 230 to 400 mesh, using methylene chloride: methanol mixed solvent (100: 1 → 10: 1)). Merck), and separated into 13 fractions, and 1.8 g of fraction 1 of these fractions was again subjected to silica gel column chromatography (silica gel 60, 230) using a hexane: ethyl acetate mixed solvent (100: 0 → 0: 100). To 400 mesh, Merck). 650 g of the combined fractions from Nos. 9 to 10 were again subjected to silica gel column chromatography (silica gel 60, 230 to 400 mesh, Merck) using a hexane: acetone mixed solvent (70: 1 → 0: 100). In the third fraction therefrom, the active ingredient (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohex 186 mg of 1-ene (named compound 2) were isolated.

Example 5. Structure Determination of Compounds

The purity of each component was evaluated using analytical TLC (analytical TLC) for the compounds isolated in Examples 3 and 4. In the state identified as a single active ingredient, various spectroscopic IR (Bio-Rad Laboratories, USA), Mass (JEOL, Japane), ¹ H-NMR (Varian, CA), ¹³ C-NMR, DEPT, HSQC, HMBC and the like were carried out to determine the structure of the compound as follows.

[1] (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohexe-1-cene

Appearance: pale yellow gum

Molecular Structure: C ₂₃ H ₂₆ O ₄

TLC phase: R _f = 0.4 (hexane: acetone = 3: 2), purple color with 10% (v / v) sulfate developing reagent.

[α] ²⁵ _D 0.00 ° (acetone, c 0.17)

UV (CHCl ₃ ) λ _max (log ε) 272 (4.0), 239 (3.9) nm

IR (liquid film) ν _max 3443, 1650, 1541, 1507, 1255, 1027 cm ^-1

EIMS m / z (%) [M] ⁺ 366 (100), 284 (10), 266 (10), 98 (15)

HREIMS m / z (%) [M] ⁺ 366.1836 (calcd. 366.4570 for C ₂₃ H ₂₆ O ₄ )

¹ H-NMR and ¹³ C-NMR see Tables 1 and 2.

[2] (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohexe-1-cene

Appearance: pale yellow solid

Molecular Structure: C ₂₄ H ₂₈ O ₄

TLC phase: R _f = 0.5 (hexane: acetone = 3: 2), purple color with 10% (v / v) sulfuric acid reagent.

[α] ²⁵ _D 0.00 ° (acetone, c 0.17)

UV (CHCl ₃ ) λ _max (log ε) 272 (3.9), 239 (3.8) nm

IR (liquid film) ν _max 1650, 1541, 1515, 1458, 1262, 1138, 1028 cm ^-1

EIMS m / z (%) [M] ⁺ 380 (100), 191 (70), 159 (95), 115 (30)

¹ H-NMR and ¹³ C-NMR see Tables 1 and 2.

¹ H Compound 1 Compound 2 One 5.89 (1H, broad doublet, J = 10.1, 2.6 Hz) 5.90 (1H, broad doublet, J = 10.0 Hz) 2 5.67 (1H, doublet of doublets, J = 10.1, 2.4 Hz) 5.68 (1H, doublet of doublets, J = 10.0, 2.6 Hz) 3 3.16 (1H, doublet of doublets, J = 8.2, 2.4 Hz) 3.18 (1H, doublet of doublets, J = 8.4, 2.6 Hz) 4 2.33 (1H, broad doublet, J = 8.2 Hz) 2.35 (1H, broad doublet, J = 8.4 Hz) 5 1.92 (1H, broad doublet, J = 12.8, 2.8 Hz) 1.66 (1H, m) 1.93 (1H, dd, J = 12.8, 3.2 Hz) 1.66 (2H, m) 6 2.21 (2H, m) 2.22 (1H, m) 1.66 (2H, m) 2' 6.67 (1H, doublet, J = 1.8 Hz) 6.70 (1H, doublet, J = 2.0 Hz) 5 ' 6.83 (1H, br s) 6.77 (2H, doublet, J = 9.1 Hz) 6 ' 6.69 (1H, doublet of doublets, J = 8.0, 1.8 Hz) 6.73 (1H, doublet of doublets, J = 9.1, 2.0 Hz) One'' 6.02 (1H, doublet of doublets, J = 16.2, 7.2 Hz) 6.02 (1H, doublet of doublets, J = 16.0, 6.8 Hz) 2'' 6.10 (1H, doublet, J = 16.2 Hz) 6.10 (1H, doublet, J = 16.0 Hz) 2''' 6.81 (1H, doublet, J = 1.4 Hz) 6.81 (1H, br s) 5 '' ' 6.77 (1H, doublet, J = 9.4 Hz) 6.77 (2H, doublet, J = 9.1 Hz) 6 '' ' 6.78 (1H, doublet of doublets, J = 9.4, 1.4 Hz) 6.78 (1H, doublet of doublets, J = 9.1, 2.4 Hz) OCH ₃ 3.87 (3H, s) 3.87 (3H, s) OCH ₃ 3.86 (3H, s) 3.86 (3H, s) OCH ₃ 3.82 (3H, s) 3.85 (3H, s) OCH ₃ 3.82 (3H, s)

¹³ C Compound 1 Compound 2 One 127.7 127.8 2 130.5 130.5 3 48.3 48.2 4 45.6 45.6 5 28.0 28.0 6 24.7 24.7 One' 137.1 137.8 2' 111.1 111.9 3 ' 146.3 147.5 4' 144.0 149.1 5 ' 114.1 111.0 6 ' 121.3 120.6 One'' 132.4 132.4 2'' 129.0 129.1 One''' 131.1 131.2 2''' 108.9 108.9 3 '' ' 149.1 148.5 4''' 148.4 148.8 5 '' ' 111.3 111.3 6 '' ' 118.9 119.0 OCH ₃ 56.1 56.1 OCH ₃ 56.1 56.1 OCH ₃ 56.0 56.0 OCH ₃ 56.0

Experimental Example 1. Measurement of cytotoxicity in vitro

Ginger extracts, solvent fractions, A549 (human lung cancer cells), Col2 (human colon cancer cells), SNU-638 (human stomach cancer cells) and HT-1080 (human fibrous tumors) of Compounds 1 and 2 obtained in Examples 1 to 4 above. Cytotoxicity) was measured using the SRB method (sulforhodamine B) as described in the literature (Lee et al., Chemico-Biol. Interact. , 115 , p215, 1998).

1-1. Cell culture

A549 (human lung cancer cells), Col2 (human colon cancer cells), SNU-638 (human gastric cancer cells), and HT-1080 (human fibroblast cells) were distributed from the Korean Cell Line Bank (KCLB), such as 10% FBS and 1% PSF. Using MEME medium containing was incubated in 37 ℃, 5% carbon dioxide incubator. Subcultures were performed once every 3 to 4 days.

1-2. Cytotoxicity of Ginger Extracts and Fractions

The effect on the cell growth of the ginger extract and solvent fractions obtained in Examples 1 and 2 was measured using the SRB method. In other words, 190 μl (5 × 10 ⁴ cells / ml) of various cell suspensions cultured in Experimental Example 1-1 were placed in a 96-well plate, and the samples 10 of Examples 1 and 2 dissolved in 10% DMSO therein. Incubated for 3 days with the addition of. 190 μl of the same cell suspension was added to at least 16 wells, followed by incubation for 30 minutes, to provide a zero-day control. The cultured cells were fixed with trichloroacetic acid (TCA), stained with SRB solution, lysed with Tris-base, and absorbance was measured at 515 nm. Using 10% DMSO as a control, the cell viability according to each test material treatment was converted according to Equation 1 below.

% Survival = {OD (Sample) -OD (Zero-Day) / OD (10% DMSO) -OD (Zero-Day)} × 100

When the control group was not treated with the sample at 100%, the value of the sample treatment group was expressed as a percentage of the control group, and each test substance treatment was calculated as an average value ± SEM of a duplicate or triple test. IC ₅₀ values were compared with each other in terms of concentration of test substance for 50% survival.

Cytotoxicity of methanol extracts, hexane fractions, chloroform fractions, n -butanol fractions and aqueous layers of ginger was found to be cytotoxic in chloroform extracts, and the cytotoxicity of chloroform extracts was IC ₅₀ 18.50 ㎍ / ml, SNU- IC _{50 was} 11.31 μg / ml. The experimental results are shown in Table 3 below.

division Cytotoxicity (IC ₅₀ : μg / ml) A549 Col2 SNU-638 Hexane > 20 > 20 > 20 chloroform 18.50 > 20 11.31 n-butanol > 20 > 20 > 20 water > 20 > 20 > 20

1-3. Cytotoxicity Measurements of Compounds 1 and 2

Cytotoxicity of Compounds 1 and 2 was measured by the same method as Experimental Example 1-2. Compounds 1 and 2 showed activity in A549, Col2, SNU-638, and HT-1080, and the experimental results are shown in Table 4 below.

division Cytotoxicity (IC ₅₀ : μg / ml) A549 Col2 SNU-638 HT-1080 Compound 1 8.4 11.2 6.6 7.8 Compound 2 4.8 5.9 3.3 6.1

Examples of the formulation of the pharmaceutical composition comprising the compounds 1 and 2 of the present invention will be described, but the present invention is not intended to be limited thereto but only to be described in detail.

Formulation Example 1 Preparation of Powder

Compound 1 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Compound 2 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Compound 2 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Compound 2 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Ginger Extract Dry Powder 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Drink

Ginger Extract Dry Powder 100mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantification

After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the ginger extract of the present invention, the compound of formula (I) isolated therefrom exhibits excellent cytotoxicity against cancer cells and inhibits the production of cancer, thus the ginger extract of the present invention or the compound isolated therefrom The composition containing may be usefully used as pharmaceuticals and nutraceuticals for the prevention and treatment of cancer diseases.

Claims (10)

Ginger ( Zingiber cassumunar Roxb.) A pharmaceutical composition for the prevention and treatment of cancer diseases containing a polar solvent soluble extract or a non-polar solvent soluble extract as an active ingredient. The pharmaceutical composition of claim 1, wherein the polar solvent soluble extract is water, a lower alcohol having C ₁ to C ₄ carbon atoms, or an extract soluble in a mixed solvent thereof. The pharmaceutical composition according to claim 2, wherein the polar solvent soluble extract is an extract soluble in methanol. The pharmaceutical composition of claim 1, wherein the nonpolar solvent soluble extract is an extract soluble in a nonpolar solvent such as hexane, ethyl acetate, and chloroform. The pharmaceutical composition according to claim 4, wherein the nonpolar solvent soluble extract is an extract soluble in chloroform. A pharmaceutical composition for the prevention and treatment of cancer diseases comprising the compound of formula (I) or a pharmacologically acceptable salt thereof as an active ingredient: (I) Where R ₁ , R ₂ , R ₃ and R ₄ are each independently -H, -OR ₅ , -SR ₅ or -NR ₅ ; R ₅ is hydrogen, lower alkyl or alkenyl group having 1 to 5 carbon atoms. The compound of formula (I) according to claim 6, wherein the compound of formula (I) is (±) -trans-3- (4-hydroxy-3-methoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl ] Cyclohexen-1 -ne and (±) -trans-3- (3,4-dimethoxyphenyl) -4-[( E ) -3,4-dimethoxystyryl] cyclohexen-one pharmaceutical composition . The cancer disease according to any one of claims 1 to 7, wherein the cancer disease is gastric cancer, colon cancer, breast cancer, lung cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or ocular melanoma, uterine cancer, ovary 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, endocrine gland 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) tumor, primary CNS lymphoma, A pharmaceutical composition comprising one or more diseases selected from spinal cord tumors, brain stem glioma, pituitary adenoma, and the like. A health functional food comprising the ginger extract of claim 1 or the compound of formula (I) of claim 6 and a food acceptable acceptable food supplement additive, which exhibit a prophylactic effect of cancer disease. The health functional food according to claim 9, which is a health beverage.