WO2014193067A1 - Composition pharmaceutique contenant, comme ingrédient actif, un composé séparé d'un extrait de laria nitida ou un sel pharmaceutiquement acceptable de celui-ci pour prévenir et traiter les cancers féminins et les symptômes de la ménopause - Google Patents

Composition pharmaceutique contenant, comme ingrédient actif, un composé séparé d'un extrait de laria nitida ou un sel pharmaceutiquement acceptable de celui-ci pour prévenir et traiter les cancers féminins et les symptômes de la ménopause Download PDF

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WO2014193067A1
WO2014193067A1 PCT/KR2014/000401 KR2014000401W WO2014193067A1 WO 2014193067 A1 WO2014193067 A1 WO 2014193067A1 KR 2014000401 W KR2014000401 W KR 2014000401W WO 2014193067 A1 WO2014193067 A1 WO 2014193067A1
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laria
cancer
food
female
nitida
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PCT/KR2014/000401
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English (en)
Korean (ko)
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송윤선
장민선
진영원
정시연
장동위
오세량
이중구
최상호
김수용
박춘길
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숙명여자대학교 산학협력단
한국생명공학연구원
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Publication of WO2014193067A1 publication Critical patent/WO2014193067A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a pharmaceutical composition for the prophylaxis and treatment of female cancer and menopausal symptoms containing the compound or pharmaceutically acceptable salt thereof isolated from Laria nitida extract as an active ingredient.
  • Women's cancers such as endometrial and breast cancers and diseases such as menopausal syndrome are associated with female hormones such as excessive or reduced estrogen and imbalance of estrogen and progesterone.
  • endometrial and ovarian cancers have increased by about five times since 1991, and the increase in the incidence of female hormone cancer is expected to continue in the future.
  • Risk factors for abnormal proliferation of the endometrium are prolonged exposure to estrogens without sufficient antagonism of progesterone, overweight, and westernized life.
  • Primary treatment for endometrial cancer is surgery, and female hormone therapy is effective for patients who have both positive estrogen or progesterone receptor after surgery and chemotherapy.
  • Excessive estrogen exposure which is not balanced with progesterone as a major cause of endometrial cancer, is known to increase endometrial cancer more than eight-fold, suggesting that megestrol, Megage TM, a progesterone receptor agonist. It is widely used.
  • megestrol side effects include high blood pressure, thrombophlebitis, and weight gain. Therefore, if a substance that can balance the efficacy of progesterone and estrogen from natural products is discovered, it may be an effective drug for endometrial cancer with reduced side effects.
  • breast cancer caused by excessive estrogen has increased significantly in Korean women over the past decade and is currently the highest among women.
  • the main causes of the increase in breast cancer are the high fat and high calorie westernized diet and obesity, late marriage and low fertility rate, avoiding lactation, and overexposure to estrogen.
  • the western region is more than three times more likely to develop breast cancer than Korean women, and the demand for breast cancer drugs is very high worldwide.
  • Breast cancer treatment is primarily performed after surgery, radiation therapy, chemotherapy, hormone therapy, etc. to prevent recurrence. Hormonal therapy is effective in lowering recurrence rates, but there is a problem that resistance is expressed after long-term administration over many years.
  • tamoxifen the first-choice drug, endometrial cancer induction and cell resistance after long-term administration have been pointed out as a major problem in many patients with metastatic cancer.
  • Anastazole, Letrazole, and Exemestane are currently used as aromatase inhibitors, which are secondary anti-hormonal drugs that have improved side effects and recurrence, but their use is limited to postmenopausal women.
  • An important aspect in the development of therapeutics for breast cancer is to first target molecules related to carcinogenesis in order to reduce toxicity, and to minimize the expression of drug resistance for long-term use and prevention of recurrence.
  • ER / PR As ER / PR positive patients make up two-thirds of all breast cancer patients, ER / PR is the best molecular target. Therefore, the development of selective estrogen receptor modulators (SERMs) and selective progesterone receptor modulators (SPRMs) with molecular targeting of ER / PR while minimizing side effects and drug resistance expression have been associated with breast cancer. This is a key goal in the development of therapeutics.
  • SERMs selective estrogen receptor modulators
  • SPRMs selective progesterone receptor modulators
  • HRT hormone replacement therapy
  • tamoxifen which is used as a breast cancer drug, acts as an ER antagonist in the breast, but acts as an ER agonist in the uterus and bone, so long-term use of tamoxifen may cause side effects of endometrial cancer.
  • a therapeutic agent or a postmenopausal symptom treatment agent for cancers in which ER is involved in carcinogenic mechanisms, such as breast cancer or endometrial cancer has been described as a substance capable of having a tissue-specific differential effect on ER, that is, selective female hormone receptor modulators (SERMs).
  • SERMs selective female hormone receptor modulators
  • pomegranate extract, soybean extract and isoflavone formulations, evening primrose oil, etc. are used as health foods for improving women's menopausal symptoms and preventing / treating osteoporosis.
  • These products, made from vegetable raw materials, are rapidly expanding in the market, giving consumers a sense of safety.
  • Plants of the genus Laria so far studied contain many lignan-based components, as well as cinnamic acid and its derivatives.
  • a representative component of a known plant of the genus Laria is lignan nordihydroguaiaretic acid (NDGA).
  • NDGA is a potent antioxidant and various biological activities are being studied.
  • tumor cell proliferation inhibitory effect is observed through accumulation of protein kinase C (PKC) and cyclic adenosine monophosphate (cAMP) and inhibition of calcium influx (Erashi M et al., Oncology 1995; 52: 150-155; Pavani M et al. Biochemical Pharmacology 1994; 48: 1935-1942).
  • PKC protein kinase C
  • cAMP cyclic adenosine monophosphate
  • NDGA receptor tyrosine kinase
  • IGF-1R insulin-like growth factor receptor
  • Patent Document 1 Republic of Korea Registered Patent Publication 10-0419121
  • Patent Document 2 Republic of Korea Patent Registration 10-0542479
  • Patent Document 3 Korean Unexamined Patent Publication 10-2004-0101694
  • Patent Document 4 Republic of Korea Patent Registration 10-0557006
  • Patent Document 5 Republic of Korea Patent Publication 10-1141194
  • Patent Document 6 Republic of Korea Patent Registration 10-1177508
  • Non-Patent Document 1 Anesini, C., Ferraro, G., Lopez, P., and Borda, E. (2001). Different intracellular signals coupled to the antiproliferative action of aqueous crude extract from Larrea divaricata Cav. and nor-dihydroguaiaretic acid on a lymphoma cell line. Phytomedicine 8, 1-7.
  • Non-Patent Document 2 Earashi, M., Noguchi, M., Kinoshita, K., and Tanaka, M. (1995). Effects of eicosanoid synthesis inhibitors on the in vitro growth and prostaglandin E and leukotriene B secretion of a human breast cancer cell line. Oncology 52, 150-155.
  • Non-Patent Document 3 Fujimoto, N., Kohta, R., and Kitamura, S. (2004). Estrogenic activity of an antioxidant, nordihydroguaiaretic acid (NDGA). Life Sci 74, 1417-1425.
  • Non-Patent Document 4 Gagnier, J.J., DeMelo, J., Boon, H., Rochon, P., and Bombardier, C. (2006). Quality of reporting of randomized controlled trials of herbal medicine interventions. Am J Med 119, 800 e801-811.
  • Non-Patent Document 5 Garreau, B., Vallette, G., Adlercreutz, H., Wahala, K., Makela, T., Benassayag, C., and Nunez, E.A. (1991). Phytoestrogens: new ligands for rat and human alpha-fetoprotein. Biochim Biophys Acta 1094, 339-345.
  • Non-Patent Document 7 Jordan, V.C. (2007). Chemoprevention of breast cancer with selective oestrogen-receptor modulators. Nat Rev Cancer 7, 46-53.
  • Non-Patent Document 8 Pavani, M., Fones, E., Oksenberg, D., Garcia, M., Hernandez, C., Cordano, G., Munoz, S., Mancilla, J., Guerrero, A. , and Ferreira, J. (1994). Inhibition of tumoral cell respiration and growth by nordihydroguaiaretic acid. Biochem Pharmacol 48, 1935-1942.
  • Non-Patent Document 9 Youngren, J.F., Gable, K., Penaranda, C., Maddux, B.A., Zavodovskaya, M., Lobo, M., Campbell, M., Kerner, J., and Goldfine, I.D. (2005). Nordihydroguaiaretic acid (NDGA) inhibits the IGF-1 and c-erbB2 / HER2 / neu receptors and suppresses growth in breast cancer cells.
  • NDGA Nordihydroguaiaretic acid
  • the present invention is to provide a new natural medicine and health functional food for the prevention and treatment of female cancer and menopausal symptoms by isolating and identifying a single component showing female hormones from Larrea nitida .
  • One of the important objects of the present invention is to provide a plant-derived material that can balance the effects of estrogen in the body while improving the side effects of conventional endometrial cancer therapeutics.
  • a pharmaceutical for the prevention and treatment of female cancers and menopausal symptoms containing any one or more selected from the group consisting of compounds represented by the following formulas (I) to (IV) and pharmaceutically acceptable salts thereof as an active ingredient
  • a composition is provided.
  • the pharmaceutical composition of the present invention may further include a compound represented by the following formula (V) or a pharmaceutically acceptable salt thereof.
  • Food composition for the prevention and improvement of female cancer and menopausal symptoms containing any one or more selected from the group consisting of a compound represented by the formula (I) to (IV) and a food acceptable salt thereof as an active ingredient
  • a compound represented by the formula (I) to (IV) containing any one or more selected from the group consisting of a compound represented by the formula (I) to (IV) and a food acceptable salt thereof as an active ingredient
  • the food composition of the present invention may further include a compound represented by the following formula (V) or a food acceptable salt thereof.
  • the present invention Larrea nitida
  • identifying and identifying single components that show female hormones in plants we provide new plant metabolism-derived single-component medicines and dietary supplements useful for the prevention and treatment of female cancer and menopausal symptoms.
  • the present invention first confirmed that single components derived from Laria nitida exhibit female hormones, and the pharmaceutical composition of the present invention containing these components occurs with the highest frequency among female cancers and its incidence rate. This may be useful for increasing breast cancer and endometrial cancer having a similar pathogenesis.
  • FIG. 3 shows the results of experiments on whether or not Raria nitida alters ERE-gene activity in MCF-7 cells transfected with Estrogen responsive element (ERE) (significantly different values from vehicle group are indicated by ***). In other words, the ES group and other values are indicated by +++ (*** ⁇ 0.001, +++ ⁇ 0.001).
  • Figure 4 is a result of analyzing the concentration-dependent ER binding force of the fraction (No. 1-8) of the extract of Laria nitida.
  • Figure 6 is the result of analyzing the concentration-dependent PR binding force of the fraction (8-12) of the extract of Laria nitida.
  • female cancer is meant to include both endometrial cancer, breast cancer, ovarian cancer, and other female genital cancers in which female hormones such as estrogen and progesterone are directly or indirectly involved in carcinogenesis.
  • menopausal symptoms is meant to include both hot flashes, hyperlipidemia, osteoporosis, venous thrombosis and atrophic vaginitis appearing in menopausal women.
  • NDGA non-hydroguaiaretic acid
  • V a compound which is a single active ingredient having estrogen efficacy in the present invention.
  • NDGA a lignan-based component
  • PDC protein kinase C
  • cAMP cyclic adenosine monophosphate
  • NDGA has been identified as one of the single components having estrogen efficacy, isolated from Laria nitida.
  • Larrea nitida is a plant of perennial and evergreen shrubs, mainly in Argentina. Folk remedies in South America are known to be used mainly for indigestion, body odor control, and menstrual control, but their efficacy has not been scientifically identified. Laria nittida used in the experiment in the present invention was sold from the overseas biological material hub center.
  • the process of confirming the effectiveness of Laria nitida and separating and identifying the components of the above (I) to (IV) from Laria nitida, which is effective for the prevention and treatment of female cancer and menopausal symptoms is as follows.
  • the pharmacological activity of the isolated fraction and the active ingredient of Laria nitida extract was tested to isolate and confirm 11 or more single components, and among these, 5 pharmacological activities were confirmed to be excellent.
  • Four of these are compounds represented by the above formulas (I) to (IV) to confirm female hormone pharmacological activity for the first time in the present invention.
  • the remaining one compound of formula (V) was NDGA, which has been confirmed to bind to estrogen receptor and its transcriptional activity in previous studies.
  • the pharmaceutical composition for the prevention and treatment of female cancer and menopausal symptoms of the present invention is effective at least one selected from the group consisting of the compounds represented by the formula (I) to (IV) and pharmaceutically acceptable salts thereof Contains as an ingredient.
  • the pharmaceutical composition of the present invention may further include a compound represented by Formula (V) or a pharmaceutically acceptable salt thereof.
  • compositions of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • suitable carriers excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • the pharmaceutical composition of the present invention may further include other pharmaceutically active ingredients or active mixtures.
  • compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and the like, oral formulations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.
  • Carriers, excipients and diluents that may be included in the composition 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.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid form preparations include at least one excipient such as starch, calcium carbonate, sucrose ( Sucrose) or lactose (Lactose), gelatin, etc. are mixed and prepared. 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.
  • 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.
  • base of the suppository Whitepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • the preferred dosage of the pharmaceutical composition of the present invention depends 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 preferred effects, it is preferable to administer at 0.0001 to 1000 mg / kg per day based on the compound represented by the formula (I) to (IV). 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 pharmaceutical composition 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.
  • excipients binders, disintegrants, lubricants, copulation agents, flavoring agents, etc. of the present invention are those described in the literature known in the art and include those having the same or similar functions.
  • Food composition for the prevention and improvement of female cancer and menopausal symptoms of the present invention is any one or more selected from the group consisting of the compounds represented by the formula (I) to (IV) and their food acceptable salts as an active ingredient Include.
  • the food composition of the present invention may further include a compound represented by the formula (V) or a food acceptable salt thereof.
  • the food composition comprises 0.0001 to 20% by weight of the compound represented by the formula (I) to (IV) in the total weight.
  • the food especially includes a dietary supplement.
  • Health functional food as defined in the present invention means a food prepared and processed using raw materials or ingredients having a useful functionality to the human body, "functional” is to control nutrients or physiology for the structure and function of the human body Ingestion is intended for the purpose of obtaining a beneficial effect on health uses such as a pharmaceutical action.
  • the health functional food may have any one form of tablets, capsules, powders, granules, liquids, and pills.
  • the food composition of the present invention may be a food composition in the form of a functional ingredient added to various foods or beverages.
  • the food for example, may be in the form of any one of beverages, powdered drinks, solids, chewing gum, tea, vitamin complexes, food additives.
  • the food composition of the present invention is an essential ingredient, except for containing any one of the components (I) to (IV), there are no special restrictions, and various flavors or natural carbohydrates, such as ordinary food or drink It may further contain various components.
  • 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.
  • natural flavoring agents such as, tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
  • the food composition of the present invention includes 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, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • the food composition of the present invention may include a flesh for preparing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is preferably selected in the range of 0 to about 20% by weight of the total weight of the food composition of the present invention.
  • hER Recombinant human ER
  • PR Recombinant human ER
  • 750 mol of each receptor protein was diluted with binding buffer and used as 10 nM.
  • the composition of the binding buffer is 10 mM Tris / pH 7.5, 10% glycerol, 1 mM DTT and 1 mg / ml BSA.
  • the assay developed by Soto et al., Uses the principle that ER-positive breast adenocarcinoma cells (MCF-7: BUS) overproliferate by enabling test substances to activate ER signaling.
  • MCF-7 breast adenocarcinoma cells
  • the relationship between the estrogen of the test substance and the cell proliferation of MCF-7: BUS is used, and the cell proliferation of 17 ⁇ -ES, which is the strongest ER ligand and has the largest cell proliferation effect, is relatively similar to that of the test substance.
  • the estrogen properties of the test substance can be evaluated.
  • the method introduced by Soto was modified and used.
  • the cell culture medium used in this test was phenol-red free media (CD-DMEM) containing charcoal treated serum to thoroughly exclude external estrogen effects.
  • CD-DMEM phenol-red free media
  • MCF-7 cells were seeded to 3 ⁇ 10 4 cells / well in a 12-well plate. After 24 hours, the medium was removed and replaced with phenol red free-DMEM medium (CD-DMEM) containing 5% charcoal dextran treated FBS containing a certain concentration of test substance.
  • CD-DMEM medium containing the test substance was prepared by diluting a concentration of test solution stock solution dissolved in DMSO 500-1000 times in the medium immediately before the experiment. The control group was added only DMSO (0.5-0.1%) CD-DMEM, the positive control was treated with 100 pM 17 ⁇ -ES. The final concentration of solvent in all experimental systems was 0.5% (v / v) or less.
  • MTT assay was performed after 18-36 hours in 37 °C, 5% CO 2 incubator to measure the number of living cells.
  • MTT [3- (4,5-dimethylthiazol-2-yl-) 2,5-diphenyltetrazolium bromide, Sigma] was treated to 1 mg / ml in culture and cultured for 4 hours, then the medium was removed. While lysis was performed, the purple MTT metabolite (crystalline) produced by living cells was dissolved. The amount of MTT metabolite dissolved in DMSO was measured at 540 nm by ELISA Plate Reader (UVmax, USA). Since absorbance is known to be proportional to the number of living cells, it is possible to quantitatively evaluate the degree of cell proliferation by the test substance.
  • MCF-7 cells known to have high ER content for ERE-luciferase assay and human breast cancer T47D cell line with intrinsically high PR expression were used for PRE-luciferase assay. Both cell lines were purchased from the American Tissue Culture Collection (USA). Twenty four hours before seeding, the cells were incubated in CD-DMEM medium, and the cells that reached about 90% confluency were seeded in 12-well plates at a concentration of about 5 ⁇ 10 5 / well. All tests were then performed in CD-DMEM medium.
  • the reporter (luciferase) gene including ERE or PRE-luciferase plasmid progesterone responsive elements (PRE) was transfected with Lipofectamine 2000 reagent (Invitrogen, USA).
  • ERE-luciferase assay 17 ⁇ -ES (1 nM) was treated as a positive control group, and ICI-182,780 (1 mM) as an antagonist control group, and various other concentrations of test substance (stock solution dissolved in DMSO were diluted in the medium.
  • MCF-7 was incubated in phenol red-free (CD-DMEM) for 2 days to measure estrogen-responsive genes.
  • CD-DMEM phenol red-free
  • ICI ER antagonist, 1 mM
  • constant Concentration test material was treated with the cell culture solution.
  • cells were trypsinized and separated from the culture vessel surface, and mRNA was extracted from the cell masses using a Qiagen RNeasy mini kit.
  • cDNA was obtained by reverse transcription of a predetermined amount of mRNA (5 mg) using an iScript cDNA synthesis kit (Bio-Rad).
  • the expression level of the gene was quantitatively measured by real-time PCR using cDNA, a pair of primers capable of recognizing the cDNA of the target gene, and a PCR SYBR green kit (Qiagen) reagent.
  • the expression level of the housekeeping gene, GAPDH was simultaneously measured and the ratio between the expression level of the gene and the expression level of the target gene was used for the final quantitative analysis.
  • Primers that recognize GAPDH include forward 5′-CTCTCTGCTCCTCCTGTTCGAC; And reverse 5'-TGAGCGATGTGGCTCGGCT.
  • TGF Transforming growth factor
  • pS2 Trefoil factor 2
  • Primers that recognize TGF-a for real-time RT PCR reactions are forward 5'-GTTTTTGGTGCAGGAGGACAA; And reverse 5'-CACAGCGTGCACCAACGT.
  • Primers that recognize pS2 are forward 5'-CGTGAAAGAC AGAATTGTGGTTTT; And reverse 5'-CGTCGAAACAGCAGCCCTTA.
  • Real-time PCR was performed at about 40 cycles (95 ° C; 30 seconds, 60 ° C; 30 seconds, 72 ° C; 30 seconds). Each gene expression curve is expressed in logarithm, and then the threshold cycle (C T ) is mathematically obtained.
  • the value obtained by substituting this value with 2-DDC T represents the relative expression level of a specific gene.
  • the expression level of the specific gene divided by the expression level of the house keeping gene was selected as the final value for the gene expression level, and the ER antagonist or agent treatment value was compared with that of the test substance.
  • the activity level of the PR target gene by the test material was similar to that of the ER target gene expression, but the expression levels of alkaline phosphatase and fatty acid synthase were detected as target genes of PR.
  • T47D an ER / PR-positive human breast adenocarcinoma cell line, was used for this test.
  • Uterotropic assay is an indirect assessment of estrogenicity by measuring the increase in uterine tissue mass induced by estrogen. The effect of uterine proliferation was measured by examining the effect of the test substance on immature ovary when intraperitoneally administered to female rats at 21 days of age. Immature female rats were used for the experiment. During the period of acclimatization, the subjects were selected to be close to the average body weight, and grouped by random method. Individual identification of animals was done by tail marking and tag by breeding box. The positive control was used to homogeneously suspend 17 ⁇ -ES in corn oil to 30 ⁇ g / 3 ml and then dilute it with corn oil step by step.
  • 17 ⁇ -ES was intraperitoneally injected (i.p.) for three days at three doses of 0.3, 3 and 30 ⁇ g / kg.
  • the dosage was 0.03 ml per 10 g body weight of immature let and solution preparation of the material was performed on the day of administration.
  • the test items to be tested in this test are weight and uterine weight. Body weight is measured for all animals immediately before administration and just before autopsy, and uterine weight is sacrificed by cervical dislocation 25 days after birth about 24 hours after the last administration, and the uterus is carefully removed to remove fat and fibrous tissue. After completely drying the water on the room, the weight of the uterus obtained from each rat was accurately measured using Mettler microbalance.
  • the active plant metabolites were identified and identified as the main single component with pharmacological effects.
  • the specific experimental method is as follows.
  • the active fraction is obtained by obtaining several fractions through Si gel column chromatography, MPLC, Sephadex LH-20, HPLC, LC-MS analysis. Fractions containing plant metabolites were subdivided and purified.
  • MCF-7 cell proliferation was increased 4.3 times when only 17 ⁇ -ES was treated as a positive control, and 1.2 times higher than vehicle group treated with 0.1% ethanol only after treatment with 10 -5 g / mL Laria nitida. It became. But Laria Nitti (10 -7 -10 -5 g / ml concentration) and 17 ⁇ -ES in combination treatment inhibited cell proliferation (1.3, 1.5 and 2.7-fold inhibition, compared to 17 ⁇ -ES at each concentration). Treatment with CI 182,780, an ER antagonist, confirmed that MCF-7 cell proliferation in this experiment was ER-mediated through the complete blockade of Laria nitida cell proliferation.
  • RBA [Ki (17 ⁇ -ES) / Ki ( Larreanitida ) ] ⁇ 100.
  • the uterine weight ratio was 1.64 when laria nitida extract was administered to sc immature rats at 100 and 500 mg / kg, which is a 0.36-fold reduction in uterine weight compared to the control group administered only corn oil. there was. In this study, it was confirmed that Laria nitida extract inhibits the growth of uterine tissues. In addition, weight gain in rats was observed during drug administration, and Laria nitida increased uterine atrophy and appetite through PR. It was predicted that.
  • Laria Nittida The concentration-dependent acute toxicity of Laria nitida extract was tested.
  • Laria Nittida The acute toxicity of the extracts was evaluated by sc injection at 50, 100, 250, 500 mg / kg for 3 days. 50, 100, 250 mg / kg and the maximum dose of 500 mg / kg were administered continuously for 3 days daily and observed for 4 days. At the maximum dose of 500 mg / kg, no physiological abnormalities were observed and the death rate was 0% because no animals died. So Laria Nittida LD 50 for acute toxicity of was evaluated as a high safety plant extract as more than 500 mg / kg, these results are shown in Table 3.
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • tablets are prepared by tableting according to a conventional method for preparing tablets.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • each component is added to the purified water to dissolve it, and lemon flavor is added, the above components are mixed, and then purified water is added and adjusted to 100 ml. do.
  • Female hormone substances of the present invention derived from natural products can be used in the field of medicines and functional foods as an effective substance for the treatment and prevention of female cancer and menopausal symptoms.

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  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

La présente invention concerne des compositions pharmaceutiques contenant, comme ingrédients actifs, des composants individuels séparés d'un extrait de Laria nitida ou des sels pharmaceutiquement acceptables de ceux-ci, pour prévenir et traiter les cancers féminins et les symptômes de la ménopause. La présente invention fournit des composants individuels séparés et identifiés à partir d'un extrait de Laria nitida et des compositions pharmaceutiques et des compositions de santé fonctionnelles contenant ceux-ci. La présente invention vérifie d'abord que les composants individuels dérivés de Laria nitida montrent des caractéristiques d'hormones femelles. Les substances hormonales femelles fournies dans la présente invention peuvent être utilisées dans des domaines de la médecine et des aliments de santé fonctionnels pour traiter et prévenir les cancers féminins et les symptômes de la ménopause.
PCT/KR2014/000401 2013-05-29 2014-01-14 Composition pharmaceutique contenant, comme ingrédient actif, un composé séparé d'un extrait de laria nitida ou un sel pharmaceutiquement acceptable de celui-ci pour prévenir et traiter les cancers féminins et les symptômes de la ménopause WO2014193067A1 (fr)

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KR1020130060942A KR101580716B1 (ko) 2013-05-29 2013-05-29 라리아 니티다 추출물로부터 분리한 화합물 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, 여성암과 갱년기 증상의 예방 및 치료용 약학적 조성물
KR10-2013-0060942 2013-05-29

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987006833A1 (fr) * 1986-05-07 1987-11-19 Chemex Pharmaceuticals, Inc. Modification d'extraits de plante obtenus de la famille des zygophyllaceae
WO1988003805A1 (fr) * 1986-11-19 1988-06-02 Chemex Pharmaceuticals, Inc. Composes pharmacologiquement actifs et leurs melanges, compositions organiques et sels metalliques
WO1995000129A1 (fr) * 1993-06-23 1995-01-05 Chemex Pharmaceuticals Inc. Traitement des maladies a resistance multiple aux anti-cancereux

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KR100419121B1 (ko) 2002-07-15 2004-02-18 주식회사 한국인삼공사 여성 갱년기 증상의 개선 기능을 갖는 생약조성물을함유하는 기능성 식품
KR100557006B1 (ko) 2003-04-01 2006-03-03 주식회사 바름인 식물성 여성 호르몬 물질인 이소플라본을 강화한 칡유산균 발효물 및 그의 제조 방법
KR20040101694A (ko) 2003-05-26 2004-12-03 (주)헬스마스터 갱년기 여성용 감마리놀렌산 가공식품
KR100542479B1 (ko) 2004-12-08 2006-01-11 한경대학교 산학협력단 석류 유래 피토에스트로겐을 함유하는 호르몬 대체 요법용피토에스트로겐 조성물
KR101141194B1 (ko) 2008-12-26 2012-07-09 (주)내츄럴엔도텍 폐경기 증상의 예방 또는 치료용 식물성에스트로겐 조성물
KR101177508B1 (ko) 2010-01-29 2012-08-28 고려대학교 산학협력단 폐경기 여성건강 예방 및 치료용 칡 추출물 조성물

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987006833A1 (fr) * 1986-05-07 1987-11-19 Chemex Pharmaceuticals, Inc. Modification d'extraits de plante obtenus de la famille des zygophyllaceae
WO1988003805A1 (fr) * 1986-11-19 1988-06-02 Chemex Pharmaceuticals, Inc. Composes pharmacologiquement actifs et leurs melanges, compositions organiques et sels metalliques
WO1995000129A1 (fr) * 1993-06-23 1995-01-05 Chemex Pharmaceuticals Inc. Traitement des maladies a resistance multiple aux anti-cancereux

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KR20140140294A (ko) 2014-12-09

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