KR100311199B1 - Composition having antimicrobial activity for propionibacterium acnes and inhibition activity for 5alpha-reductase - Google Patents

Abstract

PURPOSE: A composition having an antimicrobial activity for Propionibacterium acnes and an inhibition activity for 5alpha-reductase is provided, which composition has improved an antimicrobial activity for Propionibacterium acnes and treating and preventing activities for pimples. CONSTITUTION: The composition having an antimicrobial activity for Propionibacterium acnes and an inhibition activity for 5alpha-reductase comprises 0.005 to 10 wt.% of dried powder of Sophora Flavescens extract, in which the dried powder of Sophora Flavescens extract contains 0.05 to 12.9 wt.% of alkaloids as active ingredients for inhibiting the activity of 5alpha-reductase; the Sophora Flavescens extract is produced by dipping Sophora Flavescens in a solvent of at least one selected from purified water, methanol, ethanol, propanol, butanol, glycerol, propyleneglycol, 1,3-butyleneglycol, methylacetate, ethylacetate, benzene, hexane, diethylether and dichloromethane, and by evaporating or freeze-drying the Sophora Flavescens solution.

Description

Composition having inhibitory activity of 5α-reductase and antimicrobial activity on propionibacterium acnes

The present invention relates to a composition having an inhibitory activity of 5α-reductase and an antimicrobial action on propionibacterium acnes. More specifically, the present invention relates to a composition containing the extract of Sophora flavescens, which is excellent in fundamentally inhibiting the excessive production of sebum.

Androgenetic alopecia, acne and seborrheic dermatitis include chronic skin diseases (androgen-dependent disorders) and chronic skin diseases (Van Luu-The et al. 5; Characterization, Expression, and Immunohistochemical Localization of 5α-Reductase in Humna Skin, J. Invest.Dermatol) , 102, 221-226, 1994). Male hair loss, which can cause serious mental injury among middle-aged men, is directly related to the amount of testosterone. Recently, many studies have been reported for the prevention and treatment of hair loss through the inhibition of male hormone activity. In addition, acne and seborrheic dermatitis in chronic skin diseases are sebum-related disorders caused by excess sebum produced by male hormones. For the fundamental treatment, sebum overproduction is inhibited through the inhibition of male hormone activity. Should be suppressed.

Briefly explaining the action of male hormones in connection with the sebum production mechanism is as follows. Testosterone, one of the testosterones, is converted to dihydrotestosterone, an active male hormone, by 5 alpha-reductase, and the abnormal overaction of this hormone acts as a substance causing male hair loss. It acts as a major causative agent for the overproduction of sebum in sebocytes. Abnormally excessive production or secretion of sebum by the above mechanism will cause sebum-related diseases (acne, seborrheic dermatitis). As a result, masculine alopecia, acne and seborrheic dermatitis are due to the overproduction of dihydrotestosterone by the action of 5 alpha-reductase, and if the activity of 5 alpha-reductase is inhibited, the disease can be prevented and treated fundamentally and effectively. You can do it.

In the case of acne of the above diseases, one factor important other than excessively produced sebum plays an important role in exacerbation of the disease. If sebum produced by the above mechanism is not discharged out of the pores due to the hyperkeratosis of the pores, primary micro-comedones are formed and propionibacterium acnes (hereinafter referred to as anaerobic superficial bacteria) present in the sebum of the pores. , P. acnes) is a chronic inflammatory disease of papule (papule), pustules (pustule), cysts (cyst), nodules (nodule) develops and, if severe, scars (scar). As a result, in the case of acne, overproduction of sebum, hyperkeratosis of pores and blood. It is described as a disease caused by several complex factors such as the action of Acnes.

Many studies have been reported to inhibit the activity of 5-alpha-reductase to prevent or treat male hormone-related diseases such as acne, androgenetic alopecia and seborrheic dermatitis (Yasuro Sugimoto et al. 3; Cations Inhibit Specific Type I). 5α-Reductase Found in Human Skin, J. Invest.Dermatol, 104, 775-778, 1995), although several substances have been found, their effectiveness and practicality are small and considerable care is required for their use. In addition, in the case of acne, it is known that the use of antibiotics such as tetracycline as an antimicrobial agent for blood and acnes is likely to cause side effects such as the appearance of resistant bacteria or photosensitivity.

In view of these problems, the present inventors consider the activity of 5 alpha-reductase that is directly involved in the production of active male hormones, which are the main causes of male hormone-related diseases such as acne, male alopecia and seborrheic dermatitis. Blood is known to be a potent inhibitor of acne. As a result of systematic research on various natural products to develop a material having excellent antibacterial activity against Aknes and high safety even for a long time use, high alpha extract in facial cleanser, soap and cream composition is 5 alpha at low concentration. It has been found to be a remarkable fact that it strongly inhibits the activity of reductase and at the same time has a strong antibacterial effect on piacnes.

The present invention relates to a liquid, cream, paste or solid composition having a sebum production inhibitory action, characterized by containing 0.005 to 10% by weight of dry powder of high ginseng extract.

General properties, applications and confirmation of the ginseng used in the present invention is as follows. The scientific name is Sophora flavescenes or Sophora angustifolia, and the herbal name is Sophorae radix, which uses the root as a herbal medicine. The ginseng has the shape of columnar or conical root, 5 ~ 20cm long, 2 ~ 3cm in diameter, dark brown to yellow brown on the outer surface, clear vertical wrinkles, yellowish white, and peeled off skin. In addition, it has a unique nausea, smells bitter, and is persistent. Alkaloids, which are the most representative ingredients, include matrin, oxymatrine, and sophoranol. Flavonoids include curaridin and isohydroicaritin. have. The leaves also contain luteolin-7-glucose, and the seeds contain cytisine and the like.

Ginseng is used as a high-density chemical agent, peripheral vasoconstrictor, antipyretic analgesic, and parasitic parasite. Toxicity is 0.4g / kg, so avoid large doses (Ref. Han Dae-seok; 194-196, 1994). As a confirmation test, 0.5 g of red ginseng extract is added with 10 ml of water, boiled, filtered, and 2 ml of mayer reagent is dropped into 2 ml of the filtrate. A yellow precipitate is formed (refer to alkaloids such as martrin). Test, Korean Pharmacopoeia VI).

<Production of Ginseng Extract>

Referring to the extraction method of ginseng used in the present invention is as follows. First, the ginseng roots were washed and powdered, and then cooled for 5 days at room temperature with an extract solvent of about 10 times, and then the extract powder was dried, and the dried powder of the ginseng extract was obtained by filtration, concentration, and freeze drying. The extraction solvent used at this time is purified water, methanol, ethanol, propane, butanol, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, benzene, nucleic acid, diethyl ether and dichloromethane or It is preferable to use two or more types of mixed solvents for extraction.

<Evaluation of 5 Alpha-reductase Activity Inhibitory Activity of Ginseng Extract>

5 alpha-reductase activity inhibitory 5 alpha-reductase was used in the liver of rats (adult, Sprague-Dawley, male rat (8-10 weeks old), liver), A protein suspension was prepared. In addition, an enzyme reaction solution was prepared in a general manner. Radiolabeled testosterone ( ³ H testosterone) is used as a substrate of 5 alpha-reductase, and enzyme reaction solution: protein suspension: high ginseng extract = 70: 20: 10 for 2 minutes. After reacting in an incubator, the reaction solvent is decanted and the remaining material is dissolved in a developing solvent and developed on a thin layer chromatogram (TLC), and the amount of testosterone and dihydrotestosterone is quantified using a densitometer. Analysis was performed to determine the conversion rate of testosterone to dihydrotestosterone by 5 alpha-reductase, and to compare the results with the control (conversion rate without the addition of high ginseng extract) to 5 alpha-reductase. Inhibition rate was evaluated. The calculation method of inhibition rate evaluation is as follows.

% Inhibition = [(A-B) / hour x 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

<Blood of Gosam extract. Antibacterial Activity Test for Acnes>

The antimicrobial activity of the high ginseng extract obtained by the above method was cultured in liquid medium. After adding a certain concentration of ginseng extract to Aknes, incubated in an anaerobic culture tank. The minimum inhibitory concentration (MIC) was determined to inhibit the growth of Acnes. The minimum inhibitory concentration (MIC) range for Acnes ranged from 10 to 100 ppm.

<Isolation and Identification of Active Ingredients>

In order to search for an active ingredient that strongly inhibits 5 alpha-reductase and an active ingredient exhibiting excellent antimicrobial activity in the extract, the primary solvent extract obtained by the above method is a polar water layer and a nonpolar solvent layer (eg Solvent fractionation in diethyl ether, dichloromethane, hexane and the like) evaluated the 5 alpha-reductase inhibitory power of the active ingredient transferred to the solvent layer, respectively. As a result, the superior inhibitory power (0.001% to 100% inhibition rate) in the nonpolar solvent layer was evaluated. It was confirmed that the active ingredient indicating was implemented, p. As a result of evaluating the minimum inhibitory concentration for Acnes, it was confirmed that the antimicrobial activity was stronger in the nonpolar solvent layer (MIC: 1 to 10 ppm) than the water layer (MIC: about 200 ppm). It was confirmed that the transition to the floor.

The present study further proceeds to an over-solvent of the extract transferred to the non-polar solvent layer by the above method for isolation or separation and identification of the active ingredient exhibiting excellent 5-alpha-reductase inhibition and antimicrobial activity. Column chromatography was used to develop two-phase solvents system to separate and fractionate the active ingredients according to the polarity of each layer, and thin layer chromatography (TLC) Using 5 alpha-reductase inhibitory test and antimicrobial test by combining preparative solutions containing the same components, we identified the active ingredient showing excellent inhibitory and antimicrobial properties. The identification was performed to confirm the effective ingredients.

As a result of the identification experiment of the active ingredient by the above method, it was found that the active ingredient group that strongly inhibits 5 alpha-reductase is alkaloids such as martrin (1x10 ^-5 % to 100% inhibition). Also blood. Two types of constituents showing strong antimicrobial activity against Acnes were isolated. One is alkaloids such as matrin, allomatrine, oxymatrine, and sophoranol, and the other isoan hydrocaritine, curarinone, cura It was confirmed that they were flavonoids, such as a ridine and a sophoralfannon G. Alkaloids in the dry powder of high ginseng extract in the composition of the present invention is preferably contained 0.05 to 12.9% by weight, flavonoids preferably contain 0.005 to 11.3% by weight. At this time, it was confirmed that the flavonoids (MIC: 0.1-1 ppm) showed more excellent antimicrobial activity than the alkaloids (MIC: about 1 ppm).

5 alpha-reductase inhibition experiments and blood. The content of alkaloids such as matrin, allomatrine, oxymacrine, and sophoranol in the extract using high-performance liquid chromatography (HPLC) from the extract of Ginseng extracted by various extraction solvents used in the evaluation of antibacterial activity against Acnes As a result of the evaluation experiment, it was confirmed that the alkaloids in the dry powder of the solvent extract was in the range of 0.05 to 12.9% by weight, but the flavonoids were in the range of 0.005 to 11.3% by weight, depending on the type of the extraction solvent. It was in range.

The present inventors confirmed the active ingredient by the above method to prepare a composition such as a cleanser, soap, cream containing the extract 5 alpha-reductase activity inhibitory power and blood. The bactericidal power (cleanser, soap, cream) against acnes was evaluated. To determine the bactericidal effect on Acnes clinically. In order to evaluate the treatment effect of acne, which is a representative complex related disease of Acnes factor, a clinical experiment (washing agent) showed that the composition containing Ginseng extract was excellent in treating acne without showing side effects such as skin redness.

The present invention contains a ginseng extractol obtained by the above method to strongly inhibit the activity of 5 alpha-reductase and blood. The present invention relates to a composition having excellent antibacterial activity to Acnes, and includes face washes, soaps and creams. Each of the compositions of the present invention is based on the base of each of the general products without containing excess fat, and preferably contains 0.005 to 10% by weight of dry powder of ginseng extract with respect to the entire composition.

In the present invention, the face wash and soap contains a ginseng extract in the face wash and soap base, alpha-hydroxy acid, beta-hydroxy acid in order to increase the clinical effect on diseases such as acne, male hair loss and seborrheic dermatitis (β-hydroxy acid) or kelatolytic or exfoliating agents can be used. In addition, fragrances, pigments, fungicides, antioxidants, preservatives, moisturizers and the like can be used, and thickeners, inorganic salts, synthetic polymer materials and the like can be used for physical property improvement. In the present invention, the cream contains a ginseng extract in the base of the general oil-in-water type (O / W), and may use fragrances, pigments, antioxidants, preservatives, etc., and synthetic or natural materials may be used for physical property improvement.

The face wash, soap and cream of the present invention can be easily prepared by conventional manufacturing methods. Inhibitory activity and activity of 5 alpha-reductase according to the present invention. The composition of the composition having an antimicrobial action on Acnes is easy to apply to any of the other formulations, such as liquid, cream, paste or solid phase.

Hereinafter, although an Example and a comparative example demonstrate this invention, the technical scope of this invention is not limited to these Examples.

Example 1: Preparation and efficacy test of ginseng extract-containing cleansing composition

The face wash composition of Examples and Comparative Examples was prepared such that the mixture of the face wash base and the ginseng extract was uniformly mixed in the homomixer through stirring, heating, degassing, and cooling.

(Unit: weight%)

5-alpha-reductase activity inhibitory evaluation test

In order to evaluate the activity inhibitory activity against the 5 alpha-reductase of the cleanser composition prepared in the above Examples and Comparative Examples, 5% aqueous solution of the Examples and Comparative Examples was prepared as a test solution and 5alpha- Inhibition of reductase activity was evaluated. The inhibition rate calculation method is as follows.

% Inhibition = [(A-B) / A] x 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

5 alpha-reductase activity inhibitory effect is judged to be effective only when 80% or more result.

The activity inhibitory test results of the 5 alpha-reductase are shown in Table 1-1.

Table 1-1

blood. Bactericidal Test on Acnes

Blood of the face wash composition manufactured in the said Example and a comparative example. To evaluate antimicrobial activity against Acnes, pour into petri dishes (bacterial dish). Incubate in an anaerobic culture tank for 3 days after inoculation with acnes and bleed. The colony number (N ₁ ) of Acnes was measured, and the colony number (N ₂ ) was measured after incubating in an anaerobic culture vessel for 3 days or more by dropping an aqueous solution of 5% Examples and Comparative Examples on a petri dish. The number of colonies obtained by the above is avoided using the following method. The sterilization rate for Acnes was evaluated.

The bactericidal rate of blood and acnes should be 90% or more to be effective.

Said blood. The bactericidal test results for the Acdes are shown in Table 1-2 below.

TABLE 1-2

Evaluation of Clinical Effect of Korean Ginseng

Excellent activity inhibitory effect and blood on 5 alpha-reductase of Ginseng identified in the above experiment. In order to confirm the potent bactericidal action against Acnes in clinical trials, we tested the clinical effect on acne, a representative disease caused by the combination of the two. Men in their late teens to late twenties to determine the clinical effects of acne prevention and treatment using facial cleansers containing Gosam extract (Examples 1-3) and facial cleansers (Comparative Examples 1-2). (Experimental group: 10, control group: 10) was selected as a test subject and used three times a day for one month to evaluate the prevention and treatment of acne.

Judging method is to determine the reduction of cotton cloth by visual observation after grading a certain grade according to the condition of acne before starting the experiment.

It is judged that if the grade of acne of clinical trial subjects shows a reduction of 3 or more grades, the effect is excellent; if the decrease of 1-2 is shown, the effect is moderate; if the decrease does not appear, there is no effect; Experimental results for the prevention and treatment of acne are shown in Tables 1-3 below.

Table 1-3 (Cleanser)

In summary, the results of the experiment of the face wash, it was shown that the enzyme containing the ginseng extract inhibits the activity of 5 alpha-reductase strongly than that of the extract, blood. The bactericidal activity against Acnes was also shown to be excellent, and clinical trials showed very good acne treatment. In addition, as shown in the results above, ginseng extract inhibition and sebum production when less than 0.005% by weight. The bactericidal power against Acnes was found to be somewhat low, and when it exceeds 10% by weight, the sebum production inhibitory ability was excellent, but there were manufacturing difficulties. Therefore, the content of ginseng extract in the face wash used in the present invention is within the range of 0.005 to 10% by weight of sebum production inhibitory activity, blood. The bactericidal power and workability for Acnes were found to be very good.

B) Example 2 Preparation and Efficacy Test of Soap Composition Containing Ginseng Extract

After mixing the soap base and the ginseng extract in a mixer well, the soap composition in the form of a solid soap was prepared by extrusion, cutting and molding in a soap manufacturing machine, the extract of the ginseng is synthesized by a conventional soap manufacturing method as well as general soap. It can also be included in soaps and complex soaps.

5-alpha-reductase activity inhibitory evaluation test

In order to evaluate the inhibitory activity of 5-alpha-reductase activity of the soap compositions prepared in the above Examples and Comparative Examples, 5% aqueous solution of Examples and Comparative Examples was prepared as a test solution and 5 alpha- Inhibition of reductase activity was evaluated. The method of estimating inhibition rate is as follows.

% Inhibition = [(A-B) / A] x 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

5 alpha-reductase activity inhibitory effect is judged to be effective only when 80% or more result.

The 5 alpha-reductase activity inhibitory test results are shown in Table 2-1.

TABLE 2-1

blood. Bactericidal Test on Acnes

Blood of the soap composition prepared in the above Examples and Comparative Examples. To evaluate antimicrobial activity against Acnes, bleed into Petri Dish (bacterial dish). After inoculation with Acnes, the cells were incubated in an anaerobic culture tank for 3 days, and the colony number (N ₁ ) of blood and acne was measured. Colonies (N ₂ ) were measured after incubation in the bath. The number of colonies obtained by the above is avoided using the following method. The sterilization rate for Acnes was evaluated.

blood. Acne's sterilization rate should be more than 90% to determine the sterilization effect.

Said blood. The bactericidal force test results for Acnes are shown in Table 2-2.

Table 2-2

As shown in the table, it was found that soaps containing Gosam extract all inhibited the activity of 5-alpha-reductase more strongly than those without it. The bactericidal activity against Acnes was also shown to be excellent. In addition, 5 alpha-reductase activity inhibitory power and blood when the ginseng extract less than 0.005% by weight as shown in the above results. The bactericidal power against Acnes was found to be somewhat low, and when it exceeds 10% by weight, the bactericidal power was excellent, but there were manufacturing difficulties. Therefore, the content of ginseng extract in the soap used in the present invention is within the range of 0.005 to 10% by weight of 5 alpha-reductase activity inhibitory activity, p. The bactericidal power and workability for Acnes were found to be very good.

Example 3: Preparation and efficacy test of ginseng extract-containing cream composition

A cream composition was prepared by emulsifying a combination of a creamy base oil component, an aqueous component, a surfactant, and a ginseng extract in a mixer, degassing, filtration and cooling, and other additives such as preservatives, chelating agents, fragrances, and pigments. The formulation was prepared in oil-in-water (O / W) so that a small amount of oily component could be added. The ginseng extract of the above can be conveniently added to basic cosmetics such as foam cleansing, lotion, essence and the like and can be used as a preventive and therapeutic agent for acne.

5-alpha-reductase activity inhibitory evaluation test

In order to evaluate the inhibitory activity of 5-alpha-reductase activity of the cream compositions prepared in the above Examples and Comparative Examples, 5% of the Example and Comparative Example cream aqueous solutions were prepared as experimental solutions and the 5-alpha- Inhibition of reductase activity was evaluated. The inhibition rate calculation method is as follows.

% Inhibition = [(A-B) / A] x 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

5 alpha-reductase activity inhibitory effect is judged to be effective only when 80% or more result.

The 5 alpha-reductase activity inhibitory test results are shown in Table 3-1.

Table 3-1

blood. Bactericidal Test on Acnes

P. In order to check the bactericidal power of the cream compositions prepared in Examples and Comparative Examples. Acnes was inoculated in a petri dish (bacterial dish), and then cultured in an anaerobic culture tank for 3 days, and the number of colonies (N ₁ ) of blood and acnes was measured. Colonies (N ₂ ) were measured after incubation in an anaerobic culture vessel for 3 days or more on a dish. The number of colonies obtained by the above is avoided using the following method. The sterilization rate for Acnes was evaluated.

blood. Acne's sterilization rate is more than 90% results are judged to have a sterilization effect.

Said blood. The bactericidal force test results for Acnes are shown in Table 3-2 below.

Table 3-2

As shown in the table, the cream containing the ginseng extract was found to strongly inhibit the activity of 5 alpha-reductase than that containing no ginseng extract. The bactericidal activity against Acnes was also shown to be excellent. In addition, 5 alpha-reductase activity inhibitory power and blood when the ginseng extract less than 0.005% by weight as shown in the above results. The bactericidal power for Acnes was found to be somewhat low, and when it exceeds 10% by weight, the bactericidal power was excellent, but there were manufacturing difficulties. Therefore, the content of ginseng extract in the cream used in the present invention is within the range of 0.005 to 10% by weight of 5 alpha-reductase activity inhibitory activity, p. The bactericidal power and workability for Acnes were found to be very good.

Therefore, according to the present invention, the cleanser, soap and cream containing 0.005 to 10% by weight of ginseng extract have an excellent inhibitory activity of 5 alpha-reductase and have an excellent antibacterial effect on blood and acnes, especially 5 alpha-reductase. And blood. The two factors of Acnes are considered to be effective in preventing and treating acne, a representative disease that occurs.

Claims (6)

A liquid, cream, paste or solid composition having a sebum production inhibitory activity, characterized by containing 0.005 to 10% by weight of dry powder of high ginseng extract. The dry powder of high ginseng extract contains 0.05 to 12.9% by weight of alkaloids such as matrin, allomatrine, oxymatrine, and sophoranol as an active ingredient that inhibits the activity of 5α-reductase. Composition characterized in that. The method of claim 1, wherein the extractant is purified water, methanol, ethanol, propanol, butanol, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, benzene, hexane, diethyl ether and dichloromethane. The extract obtained by cooling the ginseng root at room temperature using one or more selected mixed solvents at room temperature, followed by evaporation or lyophilization of the solvent. The composition according to any one of claims 1, 2 and 3, wherein the composition is an agent for preventing or treating acne. (Secondary correction) The composition according to any one of items 1, 2 and 3, wherein the composition is an agent for preventing or treating hair loss or seborrheic dermatitis. The composition according to claim 1, wherein the composition has an activity inhibiting activity of 5 alpha -Reductase.