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WO2018105998A1 - Composition destinée à la prévention et au traitement de l'infertilité masculine, comprenant un composé dérivé de flavonoïdes en tant que principe actif, et son utilisation - Google Patents

Composition destinée à la prévention et au traitement de l'infertilité masculine, comprenant un composé dérivé de flavonoïdes en tant que principe actif, et son utilisation Download PDF

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Publication number
WO2018105998A1
WO2018105998A1 PCT/KR2017/014176 KR2017014176W WO2018105998A1 WO 2018105998 A1 WO2018105998 A1 WO 2018105998A1 KR 2017014176 W KR2017014176 W KR 2017014176W WO 2018105998 A1 WO2018105998 A1 WO 2018105998A1
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Prior art keywords
glucoside
quercetin
sperm
male infertility
methoxyquercetin
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PCT/KR2017/014176
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English (en)
Korean (ko)
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박종관
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전북대학교 산학협력단
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Publication of WO2018105998A1 publication Critical patent/WO2018105998A1/fr

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health

Definitions

  • the present invention relates to a composition for the prevention and treatment of male infertility containing the flavonoid derivative compound as an active ingredient and its use.
  • the present invention relates to a composition for the prevention and treatment of male infertility containing the flavonoid derivative compound as an active ingredient and its use.
  • Varicocele 19%-41% of primary infertility males and secondary male infertility males, characterized by pampiniform plexus expansion of the spermatic cord Appears in more than 80% of patients and is the leading cause of male infertility (Agarwal A, Deepinder F, Cocuzza M, Agarwal R, Short RA, Sabanegh E , et al. (2007) Efficacy of varicocelectomy in improving semen parameters: new meta-analytical approach. Urology 70, 532-538; Agarwal A, Sharma RK, Desai NR, Prabakaran S, Tavares A & Sabanegh E.
  • Surgical therapies for treating male infertility with the most commonly performed varicose veins include centuries of nonmicrosurgical approaches, laparoscopic or microsurgical varicose vein ligation in the male infertility area.
  • Varicocelectomy has been performed (Mehta A & Goldstein M. (2012) Microsurgical varicocelectomy: a review. Asian J Androl 15, 56-60.).
  • Varicocelectomy has been performed (Mehta A & Goldstein M. (2012) Microsurgical varicocelectomy: a review. Asian J Androl 15, 56-60.).
  • Despite a dramatic improvement in semen parameters in 50-80% of infertile men who underwent varicocelectomy pregnancy rates: pregnancy rates, 31-74%) (Cheng D, Zheng XM, Li SW, Yang ZW & Hu LQ.
  • Varicose veins-related including blood flow alterations in the testis, scrotal temperature elevations, abnormal hormone values, and excessive reactive oxygen species (ROS)
  • ROS reactive oxygen species
  • ROS is a byproduct of oxygen metabolism and energy production that acts as a regulator of essential physiological intracellular processes. Small amounts of ROS play an important role in regulating sperm function in the male reproductive tract. Under abnormal conditions, oxidative stress is followed when an increase in ROS levels or a decrease in antioxidant capacity occurs. Specifically, oxidative stress results in alteration of protein expression levels in sperm, eventually leading to molecular and genetic defects (Sharma R, Agarwal A, Mohanty G, Hamada AJ, Gopalan B, Willard B , et al. (2013) Proteomic analysis of human spermatozoa proteins with oxidative stress.Reprod Biol Endocrinol 11, 48.).
  • Pregnancy in normal couples ranges from 20-25% in the first month after normal sex, 75% in 6 months, and 85-90% in 1 year. Infertility is considered to be infertility in one year despite the normal marital relationship. Infertility patients account for 13-20% of married couples, and there are an estimated 1.4 million pairs of infertile couples in Korea. . Among them, infertility causes 35-50% of men, and 30% of them are caused by decreased sperm motility, and male infertility is an important disease occupying 4% of urological outpatient male patients.
  • the inventors of the present invention provides a pharmaceutical composition for the prevention and treatment of male infertility containing a herbal extract consisting of pageokcheon, earth and sand and onion (Korean Patent Registration No. 10-1481569 and PCT / WO2015 / 174636 A1); Veterinary composition for the prevention and treatment of infertility of mammals, fish or poultry containing a herbal extract consisting of pageukcheon, earth and sand and onion (Korean Patent Registration No. 10-1544783); There was a patent application for confirming the treatment effect on the male infertility of the earth and sand extract, in particular, the crude earth and sand extract extracted with ethanol (Korean Patent No. 10-2014-0102848).
  • the present inventors as part promote in vitro to investigate the effect of targeting a flavonoid derivative compound on sperm count and motility changes in the human and pig sperm (In vitro) sperm motility in follow-up studies to develop an effective treatment for male infertility It was confirmed that the samples of the present invention through efficacy evaluation experiment to increase the sperm motility in human and porcine sperm and completed the present invention.
  • An object of the present invention is to develop a new fertility overcoming technology to improve fertilization rate by improving sperm movement, and ultimately through the present invention to improve sperm movement and recovery of fertility rate.
  • the present invention provides quercetin 4'-glucoside (Spiraeoside), kaempferol 3-glucoside; Quercetin-3'-galactoside (Hyperoside) and 3-methoxyquercetin (3-methoxyquercetin; isorhamnetin), flavonoid compounds, isomers thereof, pharmaceutically acceptable thereof It provides a pharmaceutical composition for preventing and treating male infertility containing a possible salt as an active ingredient.
  • the present invention is Quercetin (Quercetin) 4'- glucoside (glucoside) (Spiraeoside: Spiraeoside, kaempferol) 3-glucoside (glucoside); Quercetin-3'-galactoside (Hyperoside) and 3-methoxyquercetin (3-methoxyquercetin; isorhamnetin), flavonoid compounds selected from the group consisting of isomers, pharmaceutically acceptable It provides a health functional food for preventing and improving male infertility containing salt as an active ingredient.
  • Male infertility as defined herein includes infertility caused by side effects of anticancer drugs such as cisplatin, vinblastin, bleomycin, spermatosis (if sperm is not present in sperm), spermosis (number of sperm) All sterility related to sperm, such as less than 20 million / ml), spermosis (less than 40% of sperm motility), malformed sperm (typically less than 40%).
  • anticancer drugs such as cisplatin, vinblastin, bleomycin, spermatosis (if sperm is not present in sperm), spermosis (number of sperm) All sterility related to sperm, such as less than 20 million / ml), spermosis (less than 40% of sperm motility), malformed sperm (typically less than 40%).
  • the compounds of the present invention can be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.
  • 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. Equal molar 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.
  • acid or alcohol eg, glycol monomethylether
  • organic acids and inorganic acids may be used as the free acid
  • hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid
  • methanesulfonic acid, p-toluenesulfonic acid, acetic acid, and trifluoroacetic acid may be used as the organic 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 (glutaric acid), glucuronic acid (glucuronic acid), aspartic acid, ascorbic acid, carbonic acid, vanic acid and hydroiodic acid may be used.
  • 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 non-soluble compound salt, and then evaporating and drying the filtrate.
  • 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 the invention include salts of acidic or basic groups which may be present in the compounds of the invention, unless otherwise indicated.
  • pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group
  • 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. It can be prepared through.
  • the compounds of the present invention are commercially available or can be isolated and obtained by the following preparation methods.
  • the compounds of the present invention can be prepared as follows. After washing and rinsing the herbal extract consisting of dried rupture cloth, earth and sand and / or onion, water, including purified water, a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol and butanol, or a solvent selected from a mixed solvent thereof, preferably water And an ultrasonic extraction method for 30 minutes to 48 hours, preferably 1 hour to 12 hours at a temperature of 30 ° C. to 150 ° C., preferably 50 ° C. to 100 ° C., after mixing the methanol mixed solvent, more preferably methanol several times.
  • the crude extract of the present invention can be obtained by filtration, concentration under reduced pressure, and drying of the extract obtained by performing hot water extraction, room temperature extraction or reflux extraction, preferably ultrasonic extraction, about 1 to 20 times, preferably 2 to 10 times. have.
  • the polar or non-polar solvent soluble extract of the present invention is about 0.0005 to 0.005 times the weight of the crude extract, preferably 10 to 90% ethanol crude extract, preferably 0.05 to 0.5 times the volume (v / w% Non-polar solvent soluble extract fractions which were added to non-polar solvents such as n-hexane, methylene chloride, ethyl acetate, etc. by performing normal fractionation process with ethyl acetate and butanol after addition of water; And polar solvent soluble extract fractions soluble in polar solvents such as butanol, water and the like.
  • Flash the non-polar solvent soluble extract fractions preferably ethyl acetate soluble fractions, which are used in non-polar solvents such as n-hexane, methylene chloride, ethyl acetate, and the like, by increasing the polarity of the mixed solvent of hexane and methylene chloride.
  • Purification using chromatography such as column chromatography, RP C18 column chromatography or silica gel open column chromatography, and Diaion HP-20 column chromatography, may be performed several times selectively to obtain and purify the compounds of the present invention, respectively. Can be.
  • the inventors of the present invention through the in vitro sperm motility efficacy test method to determine the effect on the sperm count and motility changes of human and porcine sperm in the flavonoid derivative compounds, By confirming the increase in sperm motility, the composition was found to be useful as a pharmaceutical composition, health functional food and health supplements for the prevention and treatment of male infertility.
  • the present invention provides a pharmaceutical composition and health functional food for the prevention and treatment of male infertility containing the above method and the compound obtained by the manufacturing process as an active ingredient.
  • composition of the present invention comprises 0.01 to 99% by weight of the compound relative to the total weight of the composition.
  • composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.
  • compositions comprising a compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • compositions comprising the compounds according to the invention are, but are not limited to, oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterilization, respectively, according to conventional methods.
  • Carriers, excipients, and diluents that may be formulated in the form of injectable solutions may 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, magnesium stearate and mineral oil.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one or more excipients in the compound, at least cotton, starch, calcium carbonate, sucrose. Or lactose, gelatin or the like is mixed.
  • lubricants such as magnesium styrate talc are also used.
  • Oral liquid preparations include suspending agents, liquid solutions, 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.
  • 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 compounds of the present invention depend 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, but may be appropriately selected by those skilled in the art.
  • the compound is preferably administered at 0.01 mg / kg to 10 g / kg per day, preferably at 1 mg / kg to 1 g / kg. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.
  • composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be anticipated, for example, by oral and rectal or intravenous methods.
  • the present invention also provides a sperm motility enhancer containing the compound as an active ingredient, the sperm motility enhancer is to improve the fertilization rate in vitro fertilization.
  • the present invention also provides female vaginal and vaginal cleansing agents and vaginal intercalating agents for treating male infertility of sperm motility enhancement comprising the compound.
  • the present invention also provides an in vitro fertilization (IVF) additive for in vitro fertilization comprising the compound to improve fertilization rate in vitro fertilization.
  • IVF in vitro fertilization
  • the present invention is Quercetin (Quercetin) 4'- glucoside (glucoside) (Spiraeoside: Spiraeoside, kaempferol) 3-glucoside (glucoside);
  • a flavonoid compound selected from the group consisting of Quercetin-3'-galactoside (Hyperside) and 3-methoxyquercetin (isorhamnetin), a pharmaceutically acceptable salt thereof Provided is a treatment method for treating male infertility patients, comprising administering to male infertility patients.
  • the present invention also provides a quercetin 4'-glucoside (Spiraeoside), kaempferol 3-glucoside (glucoside) for the manufacture of a medicament for treating male infertility disease;
  • a quercetin 4'-glucoside Spiraeoside
  • kaempferol 3-glucoside glucoside
  • flavonoid compounds selected from the group consisting of Quercetin-3'-galactoside (Hyperside) and 3-methoxyquercetin (, isorhamnetin), pharmaceutically acceptable salts thereof To provide.
  • the present invention is Quercetin (Quercetin) 4'- glucoside (glucoside) (Spiraeoside: Spiraeoside, kaempferol) 3-glucoside (glucoside); A flavonoid compound selected from the group consisting of Quercetin-3'-galactoside (Hyperside) and 3-methoxyquercetin (isorhamnetin), a pharmaceutically acceptable salt thereof To provide a health functional food for the prevention and improvement of male infertility containing as an active ingredient.
  • health functional food refers to physical or mental functionality by adding a compound of the present invention to a conventional specimen for preventing or ameliorating a target disease in a human body or a mammal according to Korean Health Functional Food Act No. 6723. It includes a functional food to improve the functionality such as.
  • the dietary supplement for the prevention and improvement of male infertility of the present invention comprises the compound in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition.
  • composition of the present invention can be used as a major component or additive and adjuvant in the manufacture of various dietary supplements and supplements for the purpose of preventing or treating male infertility or enhancing testicular function.
  • a pharmaceutical dosage form such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or health functional foods in the form of tea bags, leach teas, and health drinks. And processing is possible.
  • the present invention is Quercetin (Quercetin) 4'- glucoside (glucoside) (Spiraeoside: Spiraeoside, kaempferol) 3-glucoside (glucoside);
  • a flavonoid compound selected from the group consisting of Quercetin-3'-galactoside (Hyperside) and 3-methoxyquercetin (isorhamnetin), a pharmaceutically acceptable salt thereof It provides a health supplement for the prevention and improvement of male infertility containing the main ingredient.
  • the "main ingredient” is about 30 to 99 (w / w%), preferably 50 to 99 (w / w%), more preferably 70 to 99 (compound) of the compound of the present invention based on the total weight of the dietary supplement. w / w%)).
  • the health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract as an essential ingredient in the indicated proportions, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks.
  • 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 the natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
  • the compound according to the present invention when used as a food additive, the compound may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method.
  • the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, etc., includes all of the health food in the conventional sense.
  • the 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 its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • the compositions of the present invention may also contain pulp 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 generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
  • the compounds of the present invention may be added to food or beverages for the purpose of preventing male infertility.
  • the amount of the 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 added in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be.
  • Samples of the present invention were tested in human and swine sperm through an in vitro sperm motility test to evaluate the effects on the sperm count and motility changes of human and swine sperm in the flavonoid compounds of the present invention.
  • the composition is useful as a pharmaceutical composition, health functional food and health supplement food for the prevention and treatment of male infertility.
  • Figure 1 shows the effect on sperm motility in human sperm by the compound of the present invention (in the figure: KOG: kaempferol 3-O-glucoside, QG: quercetin-4-glucoside (spiraeoside) ,, CIN4; Meaning CINTM1);
  • HEP Hyperoside
  • IG Isorhamnetin-3-O-glucoside
  • KOG Kaempherol-3-O-glucoside
  • QC quercetin
  • QG means quercetin-4-glucoside (spiraeoside).
  • the dried extract was dissolved in physiological saline and mixed vigorously for 2 minutes at room temperature.
  • Dried earthenware (baekjangsaeng, Korea, Cuscuta) chinensis , Baek Jang Sang, Sounth Korea) 5L of 95% ethanol was added to 5.2Kg, and ultrasonically extracted twice for 90 minutes, filtered and concentrated under reduced pressure.
  • the concentrated filtrate was lyophilized with a freeze dryer (ScanVac, Labogene) under reduced pressure to obtain 264.3 g (4.36%) of extract powder and stored at 4 ° C.
  • the dried extract was dissolved in physiological saline and mixed vigorously for 2 minutes at room temperature.
  • the dried extract was dissolved in physiological saline and mixed vigorously for 2 minutes at room temperature.
  • CINTM1 dried paepukcheon extract obtained in the Reference Example 1, 1mg of Tosa seed extract (CINThera-2) and 10mg of onion extract (CINThera-3), respectively, by weighing a mixer (Vortex genie-2, scientific industries, USA ) were mixed with a catalyst to this takes the mixed herbal extract 21mg experimental sample (hereinafter, referred to as CINTM1).
  • Tosa extract In a manner similar to that disclosed in Korean Patent Publication No. 1014815690000, 5 L of 95% ethanol was added to 20.4 Kg of dried Tosa (baekjangsaeng), ultrasonically extracted twice for 90 minutes, filtered and concentrated under reduced pressure. The concentrated filtrate was lyophilized with a freeze dryer (ScanVac, Labogene) under reduced pressure to obtain 264.3 g (1.32%) of Tosa ethanol extract powder (hereinafter referred to as CJC) and stored at 4 ° C. Before use in the experiment, the dried extract was dissolved in physiological saline and mixed vigorously for 2 minutes at room temperature.
  • CJC Tosa ethanol extract powder
  • Onion extract After washing and drying onion (Changnyeong) well, three layers including the outer skin was added to 217g of 4L ethanol and extracted for 3 hours at 70 °C, filtered and concentrated under reduced pressure. The concentrated filtrate was lyophilized with a freeze dryer (ScanVac, Labogene) under reduced pressure to obtain 37.79 g (17.41%) of extract powder and stored at 4 ° C.
  • the compound was isolated and purified from the ethyl acetate fraction of the most active fraction, and the content of kaempferol 3-glucoside in 1 g of crude extract was determined by HPLC-.
  • Agilent 1290 HPLC (DAD) system and Shiseido UG18 (5 ⁇ m, 4.6 x 250mm ID) column chromatography were performed under the following analytical conditions to quantitatively analyze the content of the active ingredient kaempferol 3-glucoside; As a result, it contained 8.49 mg.
  • Gilson HPLC system Gilson 321 pump, 157 UV / Vis detector
  • Quercetin 4'-glucoside (spiraeoside: Sigma-Aldrich, 91802), kaempferol 3-glucoside (Sigma-Aldrich, catalog No. 68437-5MG), quercetin from the company ) -3'-galactoside (hyperoxide: Hyperoside (Wako, 087-08651) and 3-methoxyquercetin (, 3-methoxyquercetin;, isorhamnetin (Sigma-Aldrich, 17794)) were purchased respectively in distilled water It was dissolved and used as the following Experimental sample.
  • Controls include 3H buffer Hams F-10 (N6635, Sigma Chemical Co., USA) + 0.4% HAS (A1653, Sigma Chemical Co., USA) + 12 mM HEPES (H4034, Sigma Chemical Co., USA) [3H, pH 7.4], and the samples were also dissolved in 3H buffer, the pH was adjusted to 7.4, and then centrifuged (13,000 rpm, 2 min, 4 ° C.) using a centrifuge (A32010 (1), LABOGENE). Supernatant was obtained and used.
  • the percentage of motile spermatozoa (A: mean number of motile spermatozoa / B: total number of spermatozoa) ⁇ 100%
  • the sperm motility increase rate after 3 hours was calculated according to the following equation (2).
  • the percentage of motile spermatozoa 3 hours after incubation (sperm mobility 3 hours after incubation on control group-sperm mobility 3 hours after incubation in test group / sperm mobility 3 hours after incubation in control group) ⁇ 100%
  • Controls include 3H buffer Hams F-10 (N6635, Sigma Chemical Co., USA) + 0.4% HAS (A1653, Sigma Chemical Co., USA) + 12 mM HEPES (H4034, Sigma Chemical Co., USA) [3H, pH 7.4], and the samples were also dissolved in 3H buffer, the pH was adjusted to 7.4, and then centrifuged (13,000 rpm, 2 min, 4 ° C.) using a centrifuge (A32010 (1), LABOGENE). Supernatant was obtained and used.
  • the percentage of motile spermatozoa (A: mean number of motile spermatozoa / B: total number of spermatozoa) ⁇ 100%
  • the sperm motility increase rate after 3 hours was calculated according to the following equation (4).
  • the percentage of motile spermatozoa 3 hours after incubation (sperm mobility 3 hours after incubation on control group-sperm mobility 3 hours after incubation in test group / sperm mobility 3 hours after incubation in control group) ⁇ 100%
  • 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 active ingredient is dissolved in accordance with a conventional method for preparing an injection, and all ingredients are prepared in the above-described ingredient content per ampoule (2 ml).
  • each component is added to the purified water to dissolve it, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled in a brown bottle.
  • the solution is prepared by sterilization.
  • Vitamin A Acetate 70 mg
  • Vitamin E 1.0 mg
  • Vitamin B1 0.13 mg
  • Vitamin B2 0.15 mg
  • Vitamin B6 0.5 mg
  • Vitamin B12 0.2 mg
  • Vitamin C 10 mg
  • Nicotinamide 1.7 mg
  • composition ratio of the said vitamin and mineral mixture was mixed and comprised in the preferable embodiment the component suitable for a healthy food, you may change arbitrarily the compounding ratio considered that it does not deviate from the mind and range of this invention.
  • Citric Acid 1000 mg
  • Plum concentrate 2 g
  • the samples of the present invention were tested through an in vitro sperm exercise enhancing efficacy test to examine the effects of sperm count and motility changes of human and swine sperm on the compound of the present invention.
  • the composition can be usefully used in pharmaceutical compositions, health functional foods and health supplements for the prevention and treatment of male infertility.

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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne une composition destinée à la prévention et au traitement de l'infertilité masculine, comprenant un composé de flavonoïdes en tant que principe actif. Par l'intermédiaire d'une expérience de procédé d'évaluation d'effet favorisant le mouvement des spermatozoïdes in vitro destinée à étudier l'effet du composé dérivé de flavonoïdes de la présente invention sur des changements du nombre de spermatozoïdes et de la mobilité du sperme humain et porcin, il a été confirmé que les échantillons de la présente invention font preuve d'une mobilité accrue des spermatozoïdes dans l'expérience in vitro avec le sperme humain et porcin. Par conséquent, la composition peut servir à une composition pharmaceutique, un aliment fonctionnel de santé et un complément alimentaire de santé destinés à la prévention et au traitement de l'infertilité masculine.
PCT/KR2017/014176 2016-12-08 2017-12-06 Composition destinée à la prévention et au traitement de l'infertilité masculine, comprenant un composé dérivé de flavonoïdes en tant que principe actif, et son utilisation WO2018105998A1 (fr)

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KR1020160166526A KR101787007B1 (ko) 2016-12-08 2016-12-08 플라보노이드 유도체 화합물을 유효성분으로 함유하는 남성 불임증 예방 및 치료용 조성물
KR10-2016-0166526 2016-12-08

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WO2018105998A1 true WO2018105998A1 (fr) 2018-06-14

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KR102020722B1 (ko) * 2018-02-28 2019-09-11 전북대학교산학협력단 플라보노이드 유도체 및 이리도이드 유도체로 구성된 화합물 조합을 유효성분으로 함유하는 남성 호르몬장애 또는 갱년기 예방 및 치료용 조성물
CN112546060B (zh) * 2020-12-15 2021-08-31 北京中医药大学 降脂的中药单体组合物、制剂及用途

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