WO2019168324A1 - Composition for preventing and treating hormonal disorder or andropause in males, containing, as active ingredient, combination of compounds comprising flavonoid derivatives and iridoid derivatives, and use thereof - Google Patents

Abstract

The present invention relates to a composition for preventing and treating hormonal disorders or andropause in males, containing, as an active ingredient, a herbal medicine extract comprising a combination of compounds comprising flavonoid derivatives and iridoid derivatives, and a use thereof. In an experiment for measuring the effects of a combination comprising (a) monotropein, (b) kaempferol 3-glucoside and (c) quercetin 4'-glucoside (spiraeoside) on the weight of the testes by using an in vivo varicocele animal model, the weight of the left testis in a normal group to which a drug had been administered slightly increased over that of a normal group, and the weight of the left testis in a varicocele mouse group to which a drug had been administered significantly increased over that of a varicocele mouse group. In addition, it has been confirmed that a varicocele mouse group to which a drug is administered has blood testosterone and antioxidant activity-related blood SOD and GSH-Px activities that remarkably increased over those of a varicocele mouse group, and thus the present invention can be effectively used in a pharmaceutical composition, a functional health food and a health supplement for the prevention and treatment of hormonal disorders or andropause in males.

Description

A composition for preventing and treating male hormonal disorders or menopausal years, and use thereof, comprising as an active ingredient a combination of a flavonoid derivative and an iridoid derivative

The present invention relates to a composition for preventing and treating male hormonal disorders or menopausal diseases using a combination of compounds consisting of flavonoid derivatives and iridoid derivatives, and specifically, for preventing male menopausal disorders using a combination of compounds comprising flavonoid derivatives and iridoid derivatives. The present invention relates to a therapeutic composition, and more particularly, to a composition for preventing and treating male hormonal disorders or menopausal years, and a use thereof, which contains a compound combination consisting of a flavonoid derivative and an iridoid derivative as an active ingredient.

 The present invention relates to a composition for preventing and treating male menopausal disorders using a compound combination consisting of a flavonoid derivative and an iridoid derivative, and specifically, comprising a combination consisting of a compound combination consisting of a flavonoid derivative and an iridoid derivative as an active ingredient. It relates to a composition for preventing and treating male hormonal disorders or menopausal and its use.

Partial Androgen Deficiency in Aging Male (PADAM) is a process in which men suffer from general physical and mental decline due to body aging. Menopausal menopause, emotional anxiety, depression, dizziness, hot flashes, sweating, sleep disorders, vigor, poor memory, decreased work ability, and decreased libido are termed 'menopausal men' (Werner AA. The male climateric.J Am Med Assoc 1939; 112: 1441-1443). Male menopause refers to the gradual decline in masculine traits, overall vitality and mood due to the effects of lowered male hormones. Men, as well as women, develop menopause around the age of 50 and increase in frequency as they age, and as they age, the function of adrenal glands, sperm fertility, and Leydig cell function decrease, and serum testosterone decreases. Symptoms will appear. According to the ISSAM (International society for the study of the aging male), lowering male hormones leads to decreased sexual desire, lowered erections, decreased mood accompanied by depression and irritation, decreased memory, reduced body hair and skin aging, and bone density. Decrease, visceral fat increase, etc. (Korean Sexual Society, Sex Therapy Curriculum for the First Doctor, Nov. 2003). In addition to other environmental factors, male menopausal causes have been known to be supplemented with male hormones. Testosterone supplements include medicinal or topical medications, patched formulations, and injection therapy.

The treatment for menopausal complex syndrome involves male hormone (androgen) drug replacement therapy. However, such drug supplement therapy may exhibit hepatotoxicity, affect LDL metabolism, raise LDL, reduce HDL, and cause cardiovascular disease. It has also been reported that the administration of male hormones may adversely affect liver, lipid status, cardiovascular and prostate disease, and sleep and behavioral disorders (J. Impot. Res., 2002 Feb; 14 Suppl. 1: S93 8, it has been reported that male hormone therapy cannot be provided in the presence of asymptomatic or overt prostate cancer (J. Urol., 151: 54-61, 1994, and J. Urol., 163: 705-712). , 2000).

Pakrukcheon (巴戟 天, Monrinda officinalis) is a root of Pakrukcheon, which belongs to the rubiaceae, a vine, and is distributed in southern China and is known to contain vitamin C and sugars. (See Information Interpretation, Dohae Hyangjeom Dictionary, Younglim History, 1998, pp.927-928).

Seeds of Cuscuta chinensis or other related plants (Melanaceae Convolvulaceae) are resin glycosides, sugars, vitamin A, beta-carotene, gamma-carotene, teraxanthin, lutein, etc. It is known to contain. (Refer to Information, Dohae Hyangdae, Yeonglim, 1998, p882-883).

Allium cepa is the diameter of Siberia, Liliaceae onions from Altai, thiol, dimethyl disulfide and diallyl-disulfide. It is known to contain caffeic acid, ferulic acid and cinapic acid. (Information disclosure, Dohae Hyangdae, Yeonglim, 1998, pp. 155-156).

The inventors of the present invention invented a composition for the prevention and treatment of male infertility, containing mixed herbal extracts such as pageokcheon, saengsaesam and onion before the present invention (Korean Patent Nos. 10-1481569 and PCT / WO2015 / 174635 A2); Inventive composition for preventing and treating menopausal disorders and health functional food using mixed herbal extracts such as Pageokcheon, Japanese red ginseng and onion (Korean Patent No. 10-1714568); Inventive composition for preventing and treating male infertility containing a compound consisting of iridoid derivatives isolated from the mixed extract as an active ingredient (Korean Patent Nos. 10-1787006 and PCT / KR2017 / 014180), Flavonoids isolated from the mixed extract Inventive composition for preventing and treating male infertility containing a compound composed of a derivative as an active ingredient (Korean Patent Nos. 10-1787007 and PCT / KR2017 / 014176), and preventing and treating male infertility containing a combination of these compounds as an active ingredient The composition composition for invention (Korean Patent Nos. 10-1787008 and PCT / KR2017 / 014191) has been filed.

However, none of these documents discloses or teaches the combination of compounds consisting of flavonoid derivatives and iridoid derivatives for the therapeutic effect of preventing oral menopausal or menopausal.

Therefore, the present inventors, as part of a follow-up study in the test of the effect of testicular weight on the testis weight of the in vivo politic varicose veins animal model of the compounds isolated from the extract combination to develop an effective treatment for male infertility. The weight of the left testicle was slightly increased in the normal group administered with the drug compared to the normal group for the compound combination consisting of id derivatives (FIG. 1), and in the group administered with the drug in the varicose veins compared to the venous varicose group. The weight was significantly increased (FIG. 2). In addition, the group administered the drug to the varicose veins group showed significantly higher blood testosterone and blood SOD and GSH-Px activity (FIG. 3, FIG. 4) than the varicose veins group. It was confirmed that the increase was completed the present invention.

An object of the present invention is to develop a therapeutic agent for preventing hormonal disorders or menopause using an active ingredient as a specific compound combination.

In order to solve the above object, the present invention provides a pharmaceutical composition for preventing and treating male hormone disorder or menopausal period containing a compound combination consisting of flavonoid derivatives and iridoide derivatives as an active ingredient.

In another aspect, the present invention provides a health functional food for preventing and improving male hormone disorder or menopausal period containing a compound combination consisting of flavonoid derivatives and iridoid derivatives as an active ingredient.

Compound combinations consisting of flavonoid derivatives and iridoid derivatives as defined herein preferably comprise (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) Quercetin 4'-glucoside (Spiraeoside), more preferably, (a) Monotropein, (b) Camerol ( Kaempferol) 3-glucoside; And (c) a weight mixing ratio (w / w) of quercetin 4'-glucoside (Spiraeoside) is 1-100: 0.01-1: 1-100 parts by weight (w / w ), More preferably 1-50: 0.1-1: 1-50 parts by weight (w / w), even more preferably 1-30: 1: 1-30 parts by weight do.

More preferably, (a) 10-20 parts by weight of monotropein, (b) 1 part by weight of Kaempferol 3-glucoside and (c) Quercetin-4 ' -1 to 15 parts by weight of a compound combination of glucoside (Spiraeoside);

Even more preferably, (b) 18 to 25 parts by weight of Deacetyl asperulosidic acid, (b) 1 part by weight of Kaempferol 3-glucoside and (c Quercetin (Quercetin) -4'- glucoside (Spiraeoside: Spiraeoside) is characterized in that it comprises a combination of 1 to 10 parts by weight.

Testosterone disorders or menopause as defined herein include all testosterone disorders and menopause associated with hormonal changes such as depression, nervousness, fatigue, muscle strength, memory loss, concentration, osteoporosis, nocturia or sexual dysfunction symptoms, and the like.

The compounds of the present invention can 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. 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.

In this case, organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid, and 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. 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 the invention include salts of acidic or basic groups which may be present in the compounds of the invention, 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. It can be prepared through.

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

The compounds of the present invention are commercially available or can be isolated and obtained by the following preparation methods.

For example, the present invention will be described in detail below.

The compounds of the present invention can be prepared as follows. After washing and rinsing the herbal extract consisting of dried Paekcheoncheon, Japanese Silsam and / or Onion, water, including purified water, lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol and butanol, or a solvent selected from a mixed solvent thereof, preferably Ultrasonic extraction for 30 minutes to 48 hours, preferably 1 to 12 hours at a temperature of 30 ° C. to 150 ° C., preferably 50 ° C. to 100 ° C., followed by mixing several times with water and a methanol mixed solvent, more preferably methanol , Filtration, vacuum concentration, and drying of the extract obtained by repeatedly performing hot water extraction, room temperature extraction or reflux extraction, preferably ultrasonic extraction, about 1 to 20 times, preferably 2 to 10 times, to obtain crude extract of the present invention. Can be.

In addition, 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 present inventors in the measurement of the effect on the weight of the testicles using the in vivo static varicose vein animal model for the various compound combinations mean that the weight of the left testis in the normal group administered the drug compared to the normal and normal group administered the drug There was no, but slightly increased (Fig. 1), the weight of the left testis significantly increased in the group administered drug to the varicose veins compared to the varicose veins (Fig. 2). In addition, the group administered the drug to the varicose veins group showed significantly increased SOD and GSH-Px activity (Fig. 3, 4) in blood associated with testosterone and antioxidant activity in the blood than the varicose group. Pharmaceutical compositions and nutraceuticals for the prevention and treatment can be provided.

The composition of the present invention comprises 0.01 to 99% by weight of the compound relative to the total weight of the composition.

However, the 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 include, but are not limited to, 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. 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 or more excipients in the compound, at least cotton, starch, calcium carbonate, sucrose. Or lactose, gelatin or the like is mixed. In addition to simple excipients, 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. 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 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. However, for the desired effect, 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.

The 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.

In another aspect, the present invention (a) monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) administering a combination consisting of Quercetin 4'-glucoside (Spiraeoside) to a testosterone disorder or menopausal patient. It provides a treatment method.

In another aspect, the present invention (a) monotropein (monotropein), (b) Kaempferol 3-glucoside (glucoside) for the manufacture of a medicament for the treatment of male hormone disorders or menopausal diseases; And (c) Quercetin 4'-glucoside (Spiraeoside).

In addition, the present invention (a) Monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) Quercetin 4'-glucoside (Spiraeoside), which provides a health functional food for preventing and improving male hormonal disorders or menopausal as an active ingredient.

As defined herein, "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 menopausal disorders 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.

Furthermore, the composition of the present invention can be used as a major component or additive and adjuvant in the manufacture of various dietary supplements and dietary supplements for the purpose of preventing and treating male hormonal disorders or menopausal or enhancing testicular function.

In addition, for the purpose of preventing and improving male menopausal disorders, as a health functional food in the form of pharmaceutical dosage forms such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or tea bags, leaches, health drinks, etc. Manufacturing and processing are possible.

In addition, the present invention (a) Monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) Quercetin 4'-glucoside (Spiraeoside), which provides a dietary supplement for the prevention and improvement of male hormone disorder or menopausal.

The active ingredient in the dietary supplement as defined herein has an active ingredient in the range of about 30% to 99%, preferably 50% to 99%, more preferably 70% to 99% by weight of the total composition. it means.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the compound as an essential ingredient in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. 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 (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. 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.

When the compound according to the present invention is 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. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, etc., includes all of the health food in the conventional sense.

In addition to the above, 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.

In addition, the compounds of the present invention may be added to food or beverages for the purpose of preventing male hormone disorders or menopausal disorders. At this time, 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.

In addition, the present invention (a) Monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) a quercetin 4'-glucoside (Spiraeoside) comprising a combination comprising as an active ingredient a food or food additive for the prevention and improvement of male hormone disorders or menopause. do.

Items listed in the "Food Additives Code" include, for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, cinnamon acid, natural additives such as navy, licorice extract, crystalline cellulose, guar gum, L- Mixed preparations, such as a glutamate sodium preparation, a noodles addition alkali preparation, a preservative preparation, and a tar pigment preparation, are mentioned.

Functional foods containing the compounds of the present invention include bread, rice cakes, dried fruits, candy, chocolates, chewing gum, confectionery such as ice cream, ice cream products such as ice cream, ice cream powder milk, low fat milk, lactose-degraded milk Processed milk, Goat milk, Fermented milk, Buttered milk, Concentrated milk, Milk cream, Butter oil, Natural cheese, Processed cheese, Milk powder, Dairy products such as whey, Meat products, Egg products, Meat products such as hamburger Meat products such as processed meat products such as sausages, sausages, bacon, etc. Noodles, dried noodles, raw noodles, noodle soups, dehydrated noodles, refined noodles, frozen noodles, pasta noodles, fruit drinks, carbonated drinks, soy milk products , Lactic acid bacteria beverages such as yogurt, beverages such as mixed drinks, seasoning foods such as soy sauce, miso, red pepper paste, chunjang, cheongukjang, mixed sauce, vinegar, sauces, tomato ketchup, curry, and dressing Lean, shortening and pizza, but is not limited thereto.

The compound according to the present invention, which is added to foods including beverages in the process of preparing the health functional food, may appropriately be added or reduced in amount as necessary.

In addition, the compounds of the present invention may be added to food or beverages for the purpose of preventing male hormone disorders or menopausal disorders. At this time, 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.

(A) Monotropein, (b) Kaempferol 3-glucoside of the present invention; And (c) Quercetin 4'-glucoside (Spiraeoside) was used to determine the effect of testis on weight of testes using an in vivo vein varicose animal model. The weight of the left testicle was slightly increased in the normal group (Fig. 1), and the weight of the left testicle was significantly increased in the group receiving drug in the varicose veins compared to the venous varicose group (Fig. 2). In addition, the group administered the drug to the varicose veins group showed significantly increased SOD and GSH-Px activity in the blood related to testosterone and antioxidant activity (Figures 3 and 4) compared to the varicose group. It can be usefully used in pharmaceutical compositions, dietary supplements and health supplements for the prevention and treatment of.

Those skilled in the art will appreciate that various modifications and variations can be used in the compositions, uses and preparations of the invention without departing from the spirit or scope of the invention.

Although described in more detail by the following examples of the present invention, it should be understood that they are not intended to be limited to the following examples in any way of the present invention.

However, the following Examples and Experimental Examples are only illustrative in a range not limiting the scope of the present invention, the contents of the present invention is not limited by the following Examples and Experimental Examples.

Reference Example 1. Preparation Example of Single Herb Extract

(1) Preparation of Pageukcheon Extract

In a method as disclosed in Korean Patent Publication No. 10-1481569 (A), 6 liters of 95% ethanol was added to 2.4 kg of dried Pagokcheon (Dukhyeondang), extracted twice with hot water 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 248.03 g (10.3%) of Pakcheoncheon ethanol extract powder (hereinafter referred to as MOE) and stored at 4 ° C.

(2) Preparation of Seaweed Extract

In a method similar to that disclosed in Korean Patent Application Laid-Open No. 10-1481569 (A), 5l of 95% ethanol was added to 20.4Kg of dried dried Korean ginseng (Panax ginseng) and 90% sonicated twice, 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.

(3) Preparation of onion extract

Onion extract: After washing and drying the onion (Changnyeong) well, three layers including the outer husk was added to 217g of 4L ethanol and extracted at 70 ℃ for 3 hours, 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.

Reference Example 2. Preparation Example of Mixed Herbal Extract (Papukcheon: Cultivated Birds: 1: 10, Weight Ratio)

100 mg of dried paepukcheon extract, 10 mg saengsaesam extract and onion extract 100mg obtained in Reference Example 1 were weighed and mixed with a mixer (Vortex genie-2, scientific industries, USA) to obtain 210 mg of mixed herbal extracts.

Reference Example 1 Analysis of Active Compounds

1-1. Preparation of Mixed Extract

100 mg of dried paepukcheon extract, 10 mg saengsaesam extract and onion extract 100mg obtained in Reference Example 1 were weighed and mixed with a mixer (Vortex genie-2, scientific industries, USA) to obtain 210 mg of mixed herbal extracts.

1-2. Content analysis of the mixture

20 mg of the mixed extract obtained in Comparative Example 1 was dissolved in a mixed solvent of water and methanol (1: 1, v / v), and mono indicators of monotropein and deacetyl asperulosidic acid, Analysis was performed using Kaempferol 3-glucoside, quercetin 4'-glucoside (Spiraeoside), and quercetin.

Indicator components of each extract are shown in Table 1 below.

Surface Composition of Each Extract extract Pagukcheon: Seaweed Birds: Onions (10: 1: 10, weight ratio) Pagokcheon Monotropein, Deacetyl asperulosidic acid Japanese red ginseng Kaempferol 3-glucoside onion Quercetin 4'-glucoside (Spiraeoside), quercetin

HPLC analysis conditions reference UV Detector 254nm column Shiseido UG18 (4.6 x 250 mm, 5um) component of mobile phase (A) acetonitrile and (B) water with 0.1% trifluoroactic acid 0 min-10min (100% B) 10 min-40 min (100% -70% B) 40 min 50 min (70% -50% B) flow rate) 1 mL / min

As a result of analyzing the mixed extract, a standard calibration curve for each standard was obtained, and the content of each compound is shown in Table 3 below.

Content of each compound Content (mg / g) Mixture Ratio (Pap Pokcheon: Seaweed Bird: 1: 10, Weight Ratio) Monotropein (a) 6.69 ± 0.19 Deacetyl asperulosidic acid (b) 8.76 ± 0.23 Keampferol 3- O -glucoside (c) 0.41 ± 0.02 Quercetin4'- O -glucoside (d) 3.61 ± 0.08 Quercetin (e) 0.70 ± 0.02

Therefore, the activity combination of each compound was confirmed by grasping the content of each component in the combination of extracts showing activity (Paepukcheon: Haesilsaesam: onions: 10: 1: 10, weight ratio).

Example 1. Preparation of Active Compounds

Monotropein (Monotropein; Sigma-Aldrich, SMB00471), Kaempferol 3-glucoside Sigma-Aldrich, SMB00192 and Quercetin 4'-glucoside (Spira) Oxide: Spiraeoside: Sigma-Aldrich, 91802) were purchased and used as the following experimental sample.

Example 2. Preparation of Compound Combinations

(A) monotropein, (b) Kaempferol 3-glucoside obtained in the above examples; And (d) a compound combination of quercetin 4'-glucoside (Spiraeoside) in a 6.69: 0.41: 3.61 weight ratio (w / w = 16.31: 1: 8.8) It was used in the following experimental example (hereinafter referred to as 'MKS').

Experimental Example 1. Evaluation of the effect on the weight of testicles

In order to determine the effect on the weight of the testicles of the samples of the Example was carried out using the method described in the literature as follows

1-1. Experiment process

This study was conducted according to the Experimental Committee of Chonbuk National University followed Basel Declaration. The experimental animals were provided with 300 ~ 350g Sprague-Dawley rats (males), 9 weeks old, from KOATECH (Pyeongtaek, South Korea).

The kennel was adjusted to 12 hours from 7 am to 7 pm by artificial lighting. The indoor temperature was maintained at 18-23 ℃ and 40-60% humidity, and the drinking water and feed were freely eaten.

1-2. Establishing a Vein Varicose Vein Animal Model

To construct an in vivo vein varicose animal model, after an intraperitoneal anesthesia with an anesthetic (Ketamine + Xylazine ((75 ~ 80) + 2.5mg / kg, Bayer, Ansan, Korea)) and approaching the left kidney through abdominal mediastinal incision, After exposing the connected renal vein (RV) and applying a 0.85mm probe (JD-S-124, JEONG DO BIO & PLANT CO. LTD, Korea) in parallel with the left renal vein, try to ligation with 5.0 Nylon The diameter of the renal vein was reduced to 50% and the ligation of the internal spermatic vein (ISV) branch was completely ligated. An intravenous vein connecting the iliac vein (CIV) and the common iliac vein (CIV) was also ligated to confirm that the intravenous vein diameter increased after the operation of Varicocele ( Andrology 2014; 2: 466-73). Analysis of testicular-internal spermatic vein variation an d the recreation of varicocoele in a Sprague-Dawley rat model)

After confirming the induction of varicose veins, the normal control group and the varicose veins induced group was divided into two groups, one group was administered saline and the other group was orally administered MKS (200 mg / kg / day) for 28 days. Testicles were weighed after the end of MKS, and hormonal tests were performed on blood from experimental animals.

1-3. Experiment result

On the 28th day after MKS administration, the weight of the left testis was significantly increased in the MKS-administered group (VC + MKS200, 2.07 ± 0.13 g) among the varices-induced varices-induced group compared with the venous varicose surgery group. It was. The control group (CTR) was 2.01 ± 0.06g, and the varicose vein surgery group (VC) was 1.76 ± 0.36g.

The effect on Testis left weight CTR CTR + MKS VC VC + MKS Testis left weight (g) 2.01 ± 0.06 2.05 ± 0.12 1.76 ± 0.36 2.07 ± 0.13 ^* CTR: normal control group; CTR + MKS: normal rats administered MKS (Monotropein + Kaemferol-3-O-glucoside + Quercetin 4-O-glucoside, w / w = 16.31: 1: 8.8); VC: varicocele-induced rats; VC + MKS: varicocele-induced rats administered MKS. ^* Significantly different from VC group ( p <0.05).

Experimental Example 2 Evaluation of the Effect on Blood Testosterone Levels

In order to determine the effect on the testosterone level in the blood of the samples of the above example, the experiment was carried out using the method described in the literature ( Drug Des Devel Ther. 2017; 11: 2969-2979.Protective effect of DA-9401 in finasteride-induced apoptosis in rat testis: inositol requiring kinase 1 and c-Jun N-terminal kinase pathway)

2-1. biochemistry

Blood was recovered from the rat vena cava and for testosterone determination, 10 μ heparin (1 mL blood was added to 1 mL blood and centrifuged at 3500 g for 10 min.) Plasma was transferred to a 5 mL tube and paraffin Encapsulated in a paraffin film All samples were transferred to a hospital laboratory.

2-2. Experiment result

Blood testosterone levels were measured at 28 days after MKS administration. The control (CTR) was 1.52 ± 0.43 ng / ml and the varicose vein surgery group (VC) was 0.93 ± 0.36 ng / ml. It was confirmed that the decrease. The blood testosterone levels in the MKS-administered group (VC + MKS200, 2.56 ± 0.64ng / ml) increased significantly compared to the varicose vein surgery group. (See Table 5)

The effect on testosterone level CTR CTR + MKS VC VC + MKS Testosterone level (ng / ml) 1.52 ± 0.43 1.47 ± 0.87 0.93 ± 0.36 2.56 ± 0.64 ^** CTR: normal control group; CTR + MKS: normal rats administered MKS (Monotropein + Kaemferol-3-O-glucoside + Quercetin 4-O-glucoside, w / w = 16.31: 1: 8.8); VC: varicocele-induced rats; VC + MKS: varicocele-induced rats administered MKS. ^** Significantly different from VC group ( p <0.001).

Experimental Example 3. Enzyme Activity Analysis

In order to confirm the antioxidant capacity of the samples of the Example was analyzed as follows using the enzyme activity method.

3-1. Determination of Glutathione Peroxidase (GSH-px) Activity

In order to confirm the antioxidant capacity of the samples of the above example, it was analyzed as follows using the method of measuring glutathione peroxidase (GSH-px) activity described in the literature (placenta . 2004; 25: 78-84. Altered placental oxidative stress status in gestational diabetes mellitus)

For the testes of the isolated rats (Han test animal, Sexually mature male SD rats, 250-300 gm and 9-10 weeks old), put into a 10-fold volume of phosphate buffered saline (PBS), homogenized and centrifuged at 400 g for 10 minutes. The supernatant was discarded and pellets were taken. 10ml of cell extraction buffer [50mM Tris-HCL, 5mM EDTA, 1mM DTT] was vortexed for 5 minutes for 30 minutes, centrifuged at 10,000g for 15 minutes, and the supernatant was used in the following experiment. GSH-px determination kit (Item No. 703102, Cayman Chemical, Ann Arbor, MI, USA) was used to measure GSH-px activity, and the measured absorbance values were calculated to indicate the production of GSH-px (nmol / mg). It was.

3-2. SOD activity measurement

In order to confirm the antioxidant capacity of the samples of the above example, it was analyzed using the method of measuring SOD activity described in the literature (placenta . 2004; 25: 78-84. Altered placental oxidative stress status in gestational diabetes mellitus).

For the testes of the isolated rats (Han test animal, Sexually mature male SD rats, 250-300 gm and 9-10 weeks old), put into a 10-fold volume of phosphate buffered saline (PBS), homogenized and centrifuged at 400 g for 10 minutes. The supernatant was discarded and pellets were taken. 10 ml of cell extraction buffer [20 mM HEPES buffer, 1 mM EGTA, 210 mM mannitol 70 mM sucrose] was vortexed for 5 minutes at 30 minutes, centrifuged at 1,500 g for 15 minutes, and the supernatant was used in the following examples. In order to measure SOD activity, a SOD determination kit (Item No. 706002, Cayman Chemical, Ann Arbor, MI, USA) was used and the measured absorbance values were calculated to indicate the amount of SOD (u / mg of protein) produced.

3-2. Experiment result

Glutathione peroxidase (GSH-px) activity was measured after MKS administration. It was confirmed that peroxidase (GSH-px) activity was decreased.

The glutathione peroxidase (GSH-px) activity of the MKS-administered group (VC + MKS200, 37.73 ± 3.92 nmol / mg) was significantly increased in the varicose vein-induced group compared to the varicose vein surgery group. (See Table 6)

The effect on glutathione peroxidase activity CTR CTR + MKS VC VC + MKS GSH-Px (nmol / mg) 33.45 ± 5.05 40.17 ± 6.76 23.82 ± 4.91 37.73 ± 3.92 ^* CTR: normal control group; CTR + MKS: normal rats administered MKS (Monotropein + Kaemferol-3-O-glucoside + Quercetin 4-O-glucoside, w / w = 16.31: 1: 8.8); VC: varicocele-induced rats; VC + MKS: varicocele-induced rats administered MKS. ^* Significantly different from VC group ( p <0.05).

As a result of measuring SOD activity after MKS administration, the control (CTR) was 8.32 ± 0.59 u / mg, and the varicose vein surgery group (VC) was 4.12 ± 0.68 u / mg. It was. The MOD-administered group (VC + MKS200, 6.11 ± 31.05 u / mg) showed significantly increased SOD activity compared to the varicose vein surgery group. (See Table 7)

The effect on SOD activity CTR CTR + MKS VC VC + MKS SOD activity (u / mg) 8.32 ± 0.59 7.03 ± 1.43 4.12 ± 0.68 ^** 6.11 ± 1.05 ^* CTR: normal control group; CTR + MKS: normal rats administered MKS (Monotropein + Kaemferol-3-O-glucoside + Quercetin 4-O-glucoside, w / w = 16.31: 1: 8.8); VC: varicocele-induced rats; VC + MKS: varicocele-induced rats administered MKS. ^* Significantly different from VC group ( p <0.05), ^** Significantly different from CTR group ( p <0.001).

Statistical analysis

Data was expressed as mean ± SD and statistical analysis was performed using software (SigmaPlot 12.0, Systat Software, San Jose, CA, USA). P <0.001 was considered statistically significant.

Hereinafter, the formulation method and the type of carrier of the present invention will be described, but the present invention is not intended to be limited thereto, but specifically, a representative formulation example will be described below.

Formulation Example 1 Preparation of Powder

MKS ------------------------------------------------- ---- 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

MKS ------------------------------------------------- ---- 10 mg

Corn Starch -------------------------------------------- 100 mg

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

Magnesium Stearate ------------------------------------- 2 mg

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

Formulation Example 3 Preparation of Capsule

MKS ------------------------------------------------- --- 10 mg

Crystalline Cellulose -------------------------------------- 3 mg

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

MKS ------------------------------------------------- --- 10 mg

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

Sterile Distilled Water for Injection ----------------------------------- 2974 mg

Na ₂ HPO ₄ , 12H2O ------------------------------------------- 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

MKS ------------------------------------------------- ---- 20 mg

Isomerized sugar ------------------------------------------------ -10 g

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

Purified water ------------------------------------------------- -Appropriate

According to the conventional method of preparing a liquid solution, 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.

Formulation Example 6 Preparation of Healthy Food

MKS ------------------------------------------------- -1000 mg

Vitamin Mixture -------------------------------------------- Proper

Vitamin A Acetate ------------------------------------- 70 μg

Vitamin E ----------------------------------------------- 1.0 Mg

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

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

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

Vitamin B12 --------------------------------------------- 0.2 μg

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

Biotin ------------------------------------------------- 10 μg

Nicotinamide ----------------------------------------- 1.7 mg

Folic Acid ------------------------------------------------- --- 50 μg

Calcium Pantothenate ------------------------------------------ 0.5 mg

Mineral mixture -------------------------------------------- Correct

Ferrous Sulfate --------------------------------------------- 1.75 Mg

Zinc Oxide ---------------------------------------------- 0.82 mg

Magnesium Carbonate ------------------------------------------ 25.3 mg

Potassium monophosphate --------------------------------------------- 15 Mg

Dibasic calcium phosphate --------------------------------------------- 55 Mg

Potassium Citrate ---------------------------------------------- 90 mg

Calcium Carbonate ----------------------------------------------- 100 Mg

Magnesium Chloride ------------------------------------------ 24.8 mg

Although the 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.

Formulation Example 7 Preparation of Healthy Drink

MKS ------------------------------------------------- ----- 1000mg

Citric Acid ------------------------------------------------- 1000 mg

oligosaccharide ------------------------------------------------- 100 g

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

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

Add purified water ------------------------------------ total 900 ml

After mixing the above components according to a conventional healthy beverage production 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 sterilization and refrigerated and then stored in the present invention For the preparation of healthy beverage compositions.

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

As described above, the present invention may be modified in various ways, and such modifications are deemed without departing from the spirit and scope of the present invention, and all such modifications are obvious to those skilled in the art to be included within the scope of the following claims. something to do.

As mentioned above, (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) Quercetin 4'-glucoside (Spiraeoside) was used to determine the effect of testis on weight of testes using an in vivo vein varicose animal model. The weight of the left testicle was slightly increased in the normal group treated with, and the weight of the left testicle was significantly increased in the group administered with the drug in the varicose veins. In addition, the group administered the drug to the varicose veins group showed significantly increased SOD and GSH-Px activity in the blood related to testosterone and antioxidant activity in the varicose veins group, thereby preventing and treating male hormone disorder or menopausal period. It can be usefully used in the composition, dietary supplements and health supplements.

Claims (12)

(a) Monotropein, (b) Kaempferol 3-glucoside; And (c) Quercetin (Quercetin) 4'- glucoside (glucoside) (Spiraeoside) containing a combination consisting of an active ingredient male hormone disorders or menopausal prevention and treatment pharmaceutical composition. According to claim 1, wherein (a) monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) a weight mixing ratio (w / w) of quercetin 4'-glucoside (Spiraeoside) is 1-100: 0.01-1: 1-100 parts by weight (w / w Pharmaceutical composition characterized in that the formulation is formulated in the compounding ratio of. According to claim 2, wherein (a) monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) the weight mixing part (w / w) of quercetin 4'-glucoside (Spiraeoside) is 1-50: 0.1-1: 1-50 parts by weight (w / Pharmaceutical composition characterized in that the formulation is formulated in the ratio of w). According to claim 3, wherein (a) monotropein (Monotropein), (b) Kaempferol 3-glucoside (glucoside); And (c) a weight blending portion (w / w) of quercetin 4'-glucoside (Spiraeoside) in a blending ratio of 1-30: 1: 1-30 parts by weight. Pharmaceutical composition characterized in that. The pharmaceutical composition of claim 1, wherein the testosterone disorder or menopausal disorder is a symptom of depression, nervousness, fatigue, muscle strength, memory loss, concentration, osteoporosis, night urination or sexual dysfunction. (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) Quercetin 4'-glucoside (Spiraeoside) containing a combination consisting of active ingredients for preventing and improving male hormone disorders or menopausal disorders. The health functional food according to claim 6, wherein the health functional food is in the form of powder, granule, tablet, capsule, pill, suspension, emulsion, syrup, tea bag, leach tea, or health beverage. (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) Quercetin 4'-glucoside (Spiraeoside) containing a combination consisting of active ingredients for preventing and improving male hormone disorders or menopausal disorders. (a) monotropein, (b) kaempferol 3-glucoside; And (c) Quercetin 4'-glucoside (Spiraeoside) comprising a combination consisting of a food ingredient for preventing and improving male hormone disorders or menopausal. (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) a quercetin 4'-glucoside (Spiraeoside) containing a combination consisting of a food ingredient for preventing and improving male hormone disorder or menopausal. (a) Monotropein, (b) Kaempferol 3-glucoside; And (c) administering a combination consisting of Quercetin 4'-glucoside (Spiraeoside) to a testosterone disorder or menopausal patient. Treatment method. (A) Monotropein, (b) Kaempferol 3-glucoside for the manufacture of a medicament for the treatment of male hormone disorder or menopausal disease; And (c) Quercetin 4'-glucoside (Spiraeoside).