KR20190092038A - A antibacterial composition containing the extracts of silk tree bark and the feminine cleanser composition comprising the same - Google Patents

Abstract

The present invention relates to a composition containing a natural extract having antibacterial activity, and a composition for alleviating or preventing vaginitis and a feminine cleanser composition comprising the same. The composition comprising an extract of Albizzia julibrissin Durazzini bark of the present invention has an effect of effectively alleviating and preventing the vaginitis by protecting the vagina from the infection of Gardnerella vaginalis and Trichomonas vaginalis. In addition, the extract of the Albizzia julibrissin Durazzini bark has no side effects on human mucous membranes, thereby being used safely in the vagina.

Description

A antibacterial composition containing the extracts of silk tree bark and the feminine cleanser composition comprising the same}

The present invention relates to a composition containing a natural extract having an antimicrobial activity and a female cleansing composition comprising the same, and more particularly, bacterial Gardnerella vaginalis and protozoan trichomonas pannial which cause female vaginitis. The present invention relates to a composition having antimicrobial activity against trichomonas vaginalis and having an activity of inhibiting the growth of these strains, a composition for improving or preventing female vaginitis, and a female detergent composition.

The vulva of a woman protects the internal genitals from external stimuli. However, because of its proximity to the anus and the humid and warm environment, it is a very good environment for pathogens to grow. Women's vaginal microbial environment plays an important role in women's health. In the vagina, strains of the genus Lactobacillus reside, which plays a role in inhibiting the growth of pathogenic strains while maintaining the acidity of 4.5 or less in the vaginal vagina by releasing lactic acid and other antimicrobial substances during growth. If the Lactobacillus strains lose control by various bacteria such as anaerobic or Candida, the possibility of vaginitis increases as the balance of the microbial environment in the vagina is broken. This vaginitis causes premature birth, low birth weight, birth, and weakened immunity. Vaginitis in women is classified into three types: bacterial infection (Bacterial vaginosis), Candida fungi (Vulvovaginal candidiasis), and trichomonas vaginitis (Trichomonas vaginitis). Bacterial vaginitis is a disease in which the acidity of the vagina increases and the microbial environment changes due to the growth of certain bacteria in the female vagina. One of the most common inflammations in women of childbearing age, usually white or off-white vaginal discharge, with increased odor.

As antibiotics used to treat bacterial vaginitis, metronidazole and clindamycin, which are susceptible to anaerobic strains, are commonly used. According to guidelines from the Centers for Disease Control (CDC), 500 mg of metronidazole should be taken orally twice a day for 7 days or bacterial clindamycin 2% cream for 7 days before bedtime. Metronidazole prior to anaerobic DNA replication, clindamycin affects protein synthesis, preventing bacteria from growing. It is known that Gardnerena varnaris has already resistant strains to metronidazole. However, the use of such antibiotics can affect the survival of beneficial bacteria, lactic acid bacteria, as well as pathogenic bacteria, so it is important to use the appropriate dose.

Vaginitis is so frequent that it is called a 'cold in women'. Therefore, for the prevention of vaginitis, there is a need for the development of a prophylactic or therapeutic agent which has no or very low antibacterial activity against vaginal bacterial flora which is generally present, and which selectively shows antibacterial activity against pathogens causing vaginitis.

Recently, in line with the increasing interest in non-toxic plant-derived natural substances, products or patents containing plant-derived ingredients in women's cleaners have been published. As a representative example, Korean Unexamined Patent Publication No. 10-0899639 discloses the preparation of a composition for treating vaginitis including pericarp extract, and in Korean Unexamined Patent Publication No. 10-2005-0005196, licorice, jihwang, peony, cheonggung, donkey, casualty, The cleaning composition containing the extract of the herbal medicine mixture consisting of phalanges and white paper as an active ingredient has been reported to have antimicrobial activity against protozoa Trichomonas protozoa, fungi Trichomonas and Haemophilus.

However, the study using the extract of the bark of the tree used in the present invention has not been reported yet.

Accordingly, the present inventors have continued research to develop new natural substances that have no antibacterial side effects, but do not affect the beneficial bacteria such as lactic acid bacteria, while having antimicrobial effects, in particular, bacteria associated with yeast infections. It is noted that bark extracts have good antibacterial effects against the harmful bacteria Gardnerella vaginalis and the protozoan Trichomonas vaginalis , which cause female vaginosis, but do not affect the beneficial bacteria at all. The present invention has been completed by discovering.

Technical problem to be solved in the present invention is not toxic to the growth of lactic acid bacteria that survive female genitalia, by maintaining the acidity in the vagina in a slightly acidic environment, Gardnerella vaginalis that is a harmful bacterium causing female vaginitis And to provide a composition that can inhibit the growth of the protozoan Trichomonas vaginalis .

In order to achieve the above object, the present invention provides a composition having an antimicrobial activity comprising a bark extract as an active ingredient.

In addition, the present invention provides a composition for improving or preventing female vaginitis comprising the composition having the antimicrobial activity.

In addition, the present invention provides a female detergent composition comprising a composition having the antimicrobial activity.

The bark extract according to the present invention inhibits the infection and growth of the harmful bacteria Gardnerella vaginalis and the protozoan Trichomonas vaginalis , which cause female vaginitis, but also on the growth of beneficial vaginal bacteria in women. It has no nutrition and no stimulus in the mucosal stimulation test, HET-CAM, which does not harm the human body in terms of safety.

Albizzia julibrissin Durazzini used in the present invention is distributed in Asia, such as China, Taiwan, Japan, Korea, and is widely used as a garden water in the yard in Korea. This is because the silk tree is a symbol of the couple's golden thread. Although the tree is less valuable as wood, flowers and bark are used for medicinal purposes. The flowering period is June ~ July, 15 ~ 20 flowers are mountain type, the upper half is red, the lower half is white, and the fruits ripen in September ~ October.

The bark of the silk tree is listed as Hailhwanpi in the Korean Herbal Medicine Standard Collection. It is used to dry bark collected in early summer. Cylindrical or semi-cylindrical, the outer side is grayish brown or brown, and the inner side is light yellowish brown or yellowish white. It is odorless, tastes mild, is steamed, and slightly irritates the tongue. In previous studies, phytochemicals such as tannins, saponins and cytosterols are known to be rich. Efficacy is known to have liver, excitement, diuresis, hookworm, antitussive, analgesic. Such bark of bark has been studied in various ways. Recently, a study on the ethanol extract of Hwanhwan P. (E. coli) has been reported to have a therapeutic effect on renal and brain cancer (Korean Patent Publication No. 2013-0022728, Korean Patent Publication No. 2012-0092283). Has been studied to induce apoptosis. In addition, bark extracts have been reported to reduce the activity of elastase, eliminate DPPH and NO free radicals, and thus apply emphysema and pulmonary hypertension.

The term "extract" of the present invention refers to a formulation prepared by squeezing the herbal medicine with a suitable leach solution and evaporating the leach solution, but not limited thereto, but the extract obtained by the extraction treatment, the dilution or concentrate of the extract, the extract It may be a dried product obtained by drying, these adjustment products, or a refined product.

The bark extract of the present invention can be extracted using a conventional extraction solvent known in the art, the extracted liquid can be used in liquid form or concentrated and / or dried. At this time, the extraction solvent is, for example, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) mixing the lower alcohol with water A solvent, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1,3-butylene glycol can be obtained as an extraction solvent. Preferably, the extraction is performed using methanol, ethanol or butane. Depending on the extraction solvent, the extraction degree and loss degree of the active ingredient of the extract may vary, so select an appropriate extraction solvent. The extraction method is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux cooling extraction, and the like.

In addition, the silk tree bark extract may be obtained through a conventional purification process in addition to the extraction method by the extraction solvent. Obtained by various additional purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). Through the fractions, the bark extract can be obtained.

According to an embodiment of the invention, about 1 to 30 times, preferably about 10 to 20 times the amount of water, C ₁ to C ₄ lower alcohols or about 1: 0.1 of the weight of the bark To 1:10, preferably a mixed solvent of water and methanol having a mixing ratio (kg / L) of 1: 0.2 to 1: 9, stirring extraction for about 0.5 to 48 hours, preferably 1 to 30 hours, and hot water Extraction method such as extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction may be used. Preferably, the extract obtained after ultrasonic extraction may be filtered, concentrated under reduced pressure or dried to obtain the extract of the present invention.

The extract extracted with the solvent may be further fractionated with a solvent selected from the group consisting of butanol, hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water and mixtures thereof.

The silk tree bark extract may be obtained by completely removing the moisture through the obtained extract liquid by concentrating and lyophilization, the silk tree bark extract from which the moisture is completely removed in the form of powder or the powder is distilled water or a common solvent Can be used after melting.

The present invention is a composition having antimicrobial activity containing the extract of bark of bark obtained by the above method as an active ingredient, Gardnerella panginalis ( G. vaginalis ) and trichomonas pannalis ( T. vaginalis ), which are the causes of female vaginitis. The present invention provides a composition having antibacterial activity.

The antimicrobial composition of the present invention is an object of treating, preventing or improving vaginitis through the antibacterial and antiprotozoal action of gardnerella panginalis ( G. vaginalis ) and trichomonas panginalis ( T. vaginalis ), the cause of female vaginitis Can be used as

In the antimicrobial composition of the present invention, the amount of bark extract may be included in an amount of 0.0001 to 20% by weight based on the total composition, and more preferably 0.001 to 10% by weight based on the total composition. 0.0001 less than% by weight, the guard four Pasteurella pants day less (G. Vaginalis), trichomonas pants day less of the effect of, the increase in the content if a too weak antibacterial effects (T. Vaginalis), and exceeds 20% by weight There is a problem in that the increase is insufficient to ensure the stability of the formulation.

The present invention provides a composition for improving or preventing female vaginitis and a female detergent composition comprising the composition having the antibacterial activity.

According to one embodiment of the present invention, the composition having the antimicrobial activity is characterized in that it is included in an amount of 1 to 20% by weight based on the total weight of each of the composition for improving or preventing female vaginitis and feminine cleaning agent composition. At this time, when the composition having antimicrobial activity is less than 1% by weight, the effect on improving women's vaginitis is too weak, and when the amount exceeds 20% by weight, the effect of increasing the content is insufficient and the stability of the formulation is not secured. There is a problem.

The composition containing the bark extract of the present invention exhibits an antimicrobial effect and has little cytotoxicity as a natural substance.

The composition having antimicrobial activity according to one embodiment of the present invention comprises a pharmaceutically acceptable carrier in addition to the bark extract. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and is preferably applied by topical application as parenteral administration, more preferably as a female detergent.

Suitable dosages of the compositions having antimicrobial activity of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to the patient. Can be prescribed. The dosage of the composition having antimicrobial activity of the present invention is in the range of 0.001-100 mg / kg on an adult basis. In addition, in the case of external preparations, it is recommended to continue the application for 1 month to 5 times a day in an amount of 1.0 to 3.0 ml on an adult basis. However, the dosage does not limit the scope of the present invention.

Compositions having an antimicrobial activity of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. It can be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup or emulsion in an oil or an aqueous medium, or may be in the form of an exir, powder, powder, granule, tablet or capsule, and may further include a dispersing or stabilizing agent.

According to one embodiment of the present invention, the composition for improving or preventing women's vaginitis and the composition for cleaning women comprising the composition having the antimicrobial activity may be applied to cleaning agents, pharmaceuticals, cosmetics, wet wipes and the like.

The cleaning agent may be formulated in a spray type, liquid type, gel type, powder, solid soap, foam type, or the like.

Meanwhile, the bark extract of the present invention is a natural substance, which is harmless to the human body, and has almost no toxicity and side effects.

It represents the composition containing the silk tree bark extract of the present invention, antimicrobial activity against trichomonas pants day lease inducing female vaginitis (T. Vaginalis) and the guard four Pasteurella pants day less anaerobic strains (G. Vaginalis), female vaginal Lactobacillus know even Phil Ruth to keep my slightly acidic environment (L. acidophilus), Lactobacillus Creative spa tooth (L. crispatus), do not affect the growth of Lactobacillus joined Lee (L. gasseri) prevention of female yeast infection It can be usefully used as a composition having efficacy. In addition, the natural extract can be used safely because there are almost no side effects on the human mucosa.

Figure 1 shows the inhibitory activity of Gardner bark nalis and Trichomonas pannalis on the bark extracts.
Figure 2 is a graph showing the growth of the bark extract of the silk tree bark for three kinds of lactic acid bacteria.
Figure 3 shows the mucosal stimulation test results by the extract of bark tree bark.

Hereinafter, examples and the like will be described in detail to help understand the present invention. However, embodiments according to the present invention can be modified in many different forms, the scope of the invention should not be construed as limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

Preparation Example 1 Preparation of Alder Bark Extract

Aspen tree bark was purchased domestically from Samhong Construction Materials Co., Ltd., and 20 times purified water of the raw material and the raw water were extracted by hot water at 100 ° C. for 15 minutes. After hot water extraction, the impurities were removed using a filter paper, and the supernatant was concentrated using a vacuum condenser, and then lyophilized and powdered. The solution was dissolved in sterile distilled water at a concentration of 100 mg / ml, and then 0.2 μm pore size. It was used in the experiment after passing through a water-soluble injection filter.

Preparation Example 1: Experimental strain

Gardnerella pannalis ( G. vaginalis , ATCC14018) and Trichomonas pannalis ( T. vaginalis , ATCC PRA-95) were distributed from the American Type Culture Collection, and Lactobacillus asidophilus ( L. acidophilus , KCTC3164). , L. crispatus (KCTC5054) and Lactobacillus gaseri ( L. gasseri , KCTC3163) were used by the Korea Institute of Bioscience and Biotechnology.

Guard four Pasteurella pants day less (G vaginalis.) Is an anaerobic culture pack in Brain Heart Infusion medium containing 10% FBS; was 37 ℃ anaerobic culture using (Anaerobic gas pack Thermo scientific), trichomonas pants day less (T. vaginalis ) were anaerobicly cultured at 37 ° C. using ATCC medium 361 containing 10% FBS. Three kinds of lactic acid bacteria were anaerobically cultured at 37 ° C. in MRS medium / agar.

Example 1: Barberry Extract is Gardnerella Pantsalis ( Gardnerella vaginalis ) And Trichomonas trousers Trichomonas vaginalis ) Impact on growth

In order to determine whether the extract of the bark extract of Preparation Example 1 has a growth inhibitory effect of the yeast infection-inducing strain, the following experiment was carried out. Before the experiment, Gardnerella vaginalis cultured in BHI agar containing 10% FBS was scraped with a loop and placed in a BHI culture solution containing 10% FBS for static culture under anaerobic conditions, and used in the experiment. The extracts were treated by concentration and inoculated to have a final bacterial count of 1 × 10 ⁶ CFU / ml, followed by anaerobic culture for 24 hours. Trichomonas vaginalis was used for the experiment by anaerobic culture in ATCC medium 361 containing 10% FBS. The extracts were treated in 96-well microplates at different concentrations, inoculated to a final cell number of 5 X 10 ⁴ cells / ml, followed by anaerobic incubation for 24 hours. An untreated group was used as a negative control, and the cultured strains were determined to have a minimum inhibitory concentration by measuring absorbance at 600 nm wavelength. The results are shown in FIG.

Figure 1 shows the inhibitory activity of Gardnerella Barnacles and Trichomonas Barnacles on the bark extracts of Albizia julibrissin. As shown here, the barberry extract exhibits a minimum inhibitory concentration of 1 mg / ml and 0.5 mg / ml for Gardnerella Baxnalis and Trichomonas Baxnalis, respectively. And it was confirmed that having a growth inhibitory effect on Trichomonas pantsnalis.

Example 2: Determination of the effect of the bark extract on the growth of lactic acid bacteria

The following experiment was carried out to determine whether the extract of bark tree bark has an effect on the growth of lactic acid bacteria, which are human beneficial bacteria. Lactobacillus asidophilus, Lactobacillus crispatus, Lactobacillus gaseri) incubated in MRS agar were anaerobicly cultured in MRS broth and used for the experiment. The extracts were treated in 96-well microplates at different concentrations and inoculated with a final bacterial count of 5 × 10 ⁶ CFU / ml, followed by anaerobic culture for 24 hours. After culturing, the strains were visually confirmed and then absorbed at each concentration at a wavelength of 600 nm. The result of comparing the growth degree of the control group and the experimental group which is no treatment group is shown in FIG. 2.

Figure 2 is a graph showing the growth of the bark extract of the silk tree bark for lactic acid bacteria. As shown here, it was judged that there was no significant difference in absorbance between the bark extract treatment group and the control group, so that it had little effect on the survival of lactic acid bacteria.

Example 3: Confirmation of mucosal irritation by the bark extract

The HET-CAM test, an in-vivo replacement test, was performed to confirm the irritation of the bark extracts of the barberry extract. HET-CAM test was carried out using a modified brown Hy-line egg sold by Pulmuone, and incubated for 9 days at 37.5 ℃, 50% humidity. The eggs were rolled automatically every hour during the incubation period. On the 9th day, the eggs were taken out of the incubator and moisturized the surface with 0.9% saline to be used for the test. The pores of the eggs were marked and the inner skin was peeled off without destroying the inner lining using a dental saw. Intensity of stimulation was determined based on the degree of bleeding, dissolution, and coagulation caused by concentrations of 0.1 N sodium hydroxide and 1% dodecyl sulfate sodium salt, which are known to cause stimulation in HET-CAM analysis. Used at a concentration of 1%. 0.3 ml of the test substance was applied to the CAM, and then observed for 5 minutes to observe the bleeding, thawing, and coagulation reactions, and compared with the treatment with the reference substance, and scored using Equation 1 below. Shown in

In reactivity, 0 represents no stimulus, 1 represents mild stimulus, 2 represents mild stimulus, and 3 represents severe stimulus.

Figure 3 shows the mucosal stimulation test results by the extract of bark tree bark. As shown here, since the reactivity was 0 under the conditions treated with the test group, the bark extract was determined to be a safe substance without irritation to the mucosa.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1: Female Cleaning Agent

1-1. Foam type

ingredient weight% Alder Bark Extract 1.00 Butylene Glycol 2.00 glycerin 1.00 Disodium ID 0.02 Fiji-60 Hydrogenated Castor Oil 1.00 Cocamidopropyl Betaine 4.00 Lauryl Glucoside 6.00 Polysorbate 20 1.00 Sodium benzoate 0.40 Caprylic Glycol 0.20 Sodium lactate 0.50 Purified water 82.88 sum 100

1-2. Gel type

ingredient weight% Alder Bark Extract 1.00 Butylene Glycol 2.00 glycerin 1.00 Disodium ID 0.02 Fiji-60 Hydrogenated Castor Oil 1.00 Cocamidopropyl Betaine 4.00 Lauryl Glucoside 6.00 Polysorbate 20 1.00 Sodium benzoate 0.40 Caprylic Glycol 0.20 Sodium lactate 0.50 Hydroxyethyl cellulose 0.7 Purified water 82.18 sum 100

Claims (7)

Composition with antimicrobial activity, characterized in that the bark extract as an active ingredient. The method of claim 1,
The activity is characterized in that the antimicrobial activity against Gadnerella vaginalis and Trichomonas panginalis ( T. vaginalis ). The method of claim 1,
The composition of the bark extract is characterized in that it is included in an amount of 0.0001 to 20% by weight based on the total weight of the composition having antibacterial activity. A female vaginitis improvement or prevention composition comprising the composition having an antimicrobial activity according to any one of claims 1 to 4 as an active ingredient. A female detergent composition comprising a composition having an antimicrobial activity according to any one of claims 1 to 4 as an active ingredient. The method of claim 5,
The female detergent composition is a female detergent composition, characterized in that for cleaning, pharmaceutical, cosmetics or wet wipes. The method of claim 6,
Detergent detergent is a female detergent composition, characterized in that the spray type, solid type, liquid type, gel type soap or foam type of detergent.

Images