KR20200063857A - A antibacterial composition containing the defatted Camellia's seed extract and the feminine cleanser composition comprising the same - Google Patents

Abstract

The present invention relates to a composition containing a natural extract with antibacterial activity, and a composition and a feminine cleanser composition for alleviating or treating female vaginitis containing the same. The composition containing a defatted Camellia japonica seed extract of the present invention protects vagina from Gardnerella vaginalis and Trichomonas vaginalis, thereby having an effect of effectively alleviate and prevent vaginitis. In addition, the defatted Camellia japonica seed extract has no side effects on human mucosa, thereby being able to be safely used for use inside the vagina.

Description

{A antibacterial composition containing the defatted Camellia's seed extract and the feminine cleanser composition comprising the same}

The present invention relates to a composition containing a natural extract having antibacterial activity and a female cleansing agent composition comprising the same, more specifically, bacterial gardnerella vaginalis and protozoan trichomonas barnacle causing female vaginitis It relates to a composition having an antibacterial activity against Trichomonas vaginalis and an activity for inhibiting the growth of these strains, and a composition for improving or preventing female vaginitis, and a composition for cleaning women.

The female vulva protects the internal genital organs from external stimuli. However, due to the humid and warm environment close to the anus, it is a very good environment for pathogens to grow. The microbial environment in women's vagina plays an important role in women's health. Strains of the genus Lactobacillus reside inside the vagina, and their role is to suppress the growth of pathogenic strains while maintaining an acidity of 4.5 or less in the vagina by releasing lactic acid and other antibacterial substances in the growth process. When the strains of the genus Lactobacillus lose the initiative by various bacteria such as anaerobic or Candida, the possibility of infection with vaginitis increases as the balance of the microbial environment in the vagina is broken. These vaginitis causes premature birth, childbirth and low birth weight, and weakens immunity. There are three main types of vaginitis in women: Bacterial vaginosis due to bacterial infection, Vulvovaginal candidiasis, and Trichomonas vaginitis. Bacterial vaginitis is a disease in which the acidity of the vagina increases and the microbial environment changes due to the proliferation of specific bacteria in the female vagina. One of the most common inflammations of women of childbearing age, usually vaginal discharge of white or off-white color and increased odor.

As antibiotics used to treat such bacterial vaginosis, metronidazole and clindamycin, which are susceptible to anaerobic strains, are commonly used. According to the guidelines of the American Centers for Disease Control and Prevention (CDC), 500 mg of metronidazole is administered orally to bacterial vaginosis twice a day for 7 days, or to use clindamycin 2% cream once for 7 days before bedtime. Metronidazole is an anaerobic DNA replication mechanism, and clindamycin affects protein synthesis, preventing bacteria from multiplying. In the case of Gardnerna barnalis, it is known that strains resistant to metronidazole already exist. However, it is important to use these antibiotics at an appropriate dose because they can affect not only pathogenic bacteria but also the survival of beneficial bacteria, lactic acid bacteria.

Vaginitis is often called a'female cold' so that it recurs frequently. Therefore, for the prevention of vaginitis, there is no or very low antibacterial activity for the vaginal bacterial flora, which is generally present, and it is necessary to develop a preventive or therapeutic agent that selectively exhibits antibacterial activity against pathogens causing vaginitis.

Recently, according to the tendency of increasing interest in non-toxic natural substances derived from plants, products or patents containing plant-derived ingredients in female detergents have been published. As a representative example, Korean Patent Publication No. 10-0899639 published a preparation for the treatment of vaginitis containing the extract of Pork Chop, and Korean Patent Publication No. 10-2005-0005196 published Licorice, Chihwang, Peony, Cheongung, Angelica, Casualty, The cleaning agent composition containing the extract of the herbal mixture consisting of phalanx and white paper as an active ingredient is said to have antibacterial activity against Trichomonas protozoan, the fungus Trichomonas, and Haemophilus.

On the other hand, camellia (Camellia japonica L.) is a kind of tea tree and is an evergreen broad-leaved small tree growing in the southern region of Korea. Camellia seeds are spindle-shaped dark brown seeds, contained in camellia fruits, usually about 3 seeds from a single fruit. It is mainly used for ornamental purposes, and camellia seeds were used for medicinal purposes by milking with oil. Currently, it is widely used not only for medicinal use, but also for cosmetics. The residue left after steaming the camellia seeds is called camellia oil. In relation to the camellia seed extract, Korean Patent Publication No. 10-2013-0122070 discloses that the camellia seed extract and camellia oil extract can restore damaged hair and provide an improvement effect of the hair. However, there are no prior reports related to the improvement of female vaginitis by camellia oil extract.

As a result, the present inventors continued to study to develop new natural substances that have no bacterial side effects without affecting beneficial bacteria such as lactic acid bacteria while having a bactericidal infection control effect against bacteria related to antibacterial, especially female vaginitis, Camellia oil extract has excellent antibacterial effects against Gardnerella vaginalis , a harmful bacterium that causes female vaginitis, and Trichomonas vaginalis , a protozoan, but does not affect beneficial bacteria at all. Thus, the present invention was completed.

The technical problem to be solved in the present invention is toxic to the growth of lactic acid bacteria surviving in the female genitalia, and by maintaining the acidity in the vagina in a weakly acidic environment, Gardnerella vaginalis , a harmful bacterium that causes female vaginitis. And to provide a composition capable of inhibiting the growth of the protozoan Trichomonas vaginalis .

In order to solve the above technical problem, the present invention provides a composition having an antibacterial activity comprising a camellia oil extract as an active ingredient.

In addition, the present invention provides a composition for improving or preventing vaginitis in women, including the composition having the antibacterial activity.

In addition, the present invention provides a female cleanser composition comprising the composition having the antibacterial activity.

Camellia oil extract according to the present invention inhibits the infection and growth of Gardnerella vaginalis , a harmful bacterium that causes female vaginitis, and Trichomonas vaginalis , a protozoan, while nourishing the growth of beneficial bacteria in the female vagina In the HET-CAM test, which is a mucosal stimulation test, there was no irritation, and safety was not harmed.

Camellia oil used in the present invention refers to the remaining residue of the camellia seed oil.

The term "extract" of the present invention refers to a formulation in which a crude drug is squeezed into an appropriate leach solution and concentrated by evaporating the leach solution, but is not limited thereto, but an extract obtained by an extraction treatment, a dilution or concentrate of the extract, or an extract It may be a dried product obtained by drying, a crude product or a purified product thereof.

Camellia oil extract of the present invention can be extracted using conventional extraction solvents known in the art, and 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 1-4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) mixing of the lower alcohol and water Solvent, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1,3-butylene glycol can be obtained as an extraction solvent. Preferably, 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 differ, so select an appropriate extraction solvent to use. The extraction method is not particularly limited, for example, cold immersion extraction, ultrasonic extraction, reflux cooling extraction, and the like.

In addition, the camellia oil extract can be obtained through a conventional purification process in addition to the extraction method by the extraction solvent. Obtained through various additional purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. Camellia oil extract can also be obtained through fractionation.

According to an embodiment of the present invention, the camellia oil extract is about 0.5 to 20 times the weight of the camellia oil sample remaining after squeezing the oil, preferably about 1 to 15 times the amount of lower alcohol of C1 to C4, such as water, ethanol, methanol, etc. Or a mixed solvent of water and methanol having a mixing ratio (kg/L) of about 1:0.1 to 1:10, preferably 1:0.2 to 1:9, at room temperature for about 0.5 to 48 hours, preferably 1 to Using an extraction method such as stirring extraction for 30 hours, hot water extraction, cold needle extraction, reflux cooling extraction, or ultrasonic extraction, preferably, the extract obtained after hot water extraction is filtered, concentrated under reduced pressure, or dried to obtain an extract of the present invention. Can be.

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

The camellia oil extract may be one in which the obtained extract is concentrated and freeze-dried to completely remove moisture, and the camellia oil extract completely removed from the water is used in powder form or the powder is dissolved in distilled water or a common solvent. Can be used.

The present invention is a composition having an antimicrobial activity containing the camellia oil extract obtained by the above method as an active ingredient, the cause of female vaginitis gardnerella pants ( G. vaginalis ) and trichomonas pants ( T. vaginalis ) To provide a composition having antibacterial activity.

The antimicrobial composition of the present invention is the purpose of treating, preventing or improving vaginitis through antibacterial and antiprotozoal action of gardnerella pantsinalis ( G. vaginalis ) and trichomonas pantsinalis ( T. vaginalis ), which are the causes of female vaginitis. Can be used as

In the antimicrobial composition of the present invention, the amount of camellia oil extract is 0.001 to 20% by weight relative to the total composition, more preferably, the camellia oil extract can contain 0.001 to 10% by weight relative to the total composition. If it is less than 0.0001% by weight, the antimicrobial effect of Gardnerella pantsinalis ( G. vaginalis ) and Trichomonas pantsinalis ( T. vaginalis ) is too weak, and when it exceeds 20% by weight, the effect of increasing the content There is a problem that the stability in the formulation is not secured due to insufficient increase.

The present invention provides a composition for improving or preventing vaginitis in women, and a composition for cleaning women, including a composition having the antibacterial activity.

According to one embodiment of the present invention, the composition having antibacterial activity is characterized in that it is included in an amount of 1 to 20% by weight relative to the total weight of each composition for improving or preventing female vaginitis and female cleansing agents. At this time, if the composition having antibacterial activity is less than 1% by weight, the effect on improving female vaginitis is too weak, and when it exceeds 20% by weight, the increase in the effect due to the increase in content is insufficient, so that stability in the formulation is not secured. There is a problem.

The composition containing the camellia oil extract of the present invention exhibits an antibacterial effect, and has little cytotoxicity as a natural substance.

The composition having antibacterial activity according to one embodiment of the present invention includes a camellia oil extract in addition to a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is commonly used in formulation, 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, but is not limited thereto It does not work. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. 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, preferably parenterally administered, more preferably applied as a female cleansing agent in a topical application manner.

Suitable dosages of the compositions having antimicrobial activity of the present invention vary by factors such as formulation method, mode of administration, patient's age, weight, gender, morbidity, food, time of administration, route of administration, rate of excretion and response sensitivity. Can be prescribed. The dosage of the composition having antibacterial 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 an external preparation, it is good to apply for 1 to 5 times a day in an amount of 1.0 to 3.0 ml on an adult basis and continue for at least 1 month. However, the dosage is not intended to limit the scope of the present invention.

The composition having the antibacterial activity of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by those skilled in the art, or It can be manufactured by incorporating 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 aqueous medium, or may be in the form of exirs, powders, granules, tablets or capsules, and may further include dispersants or stabilizers.

According to one embodiment of the present invention, the composition for improving or preventing female vaginitis, including the composition having the antibacterial activity, and the female cleansing agent composition may be applied as a detergent, a pharmaceutical, a cosmetic, or a wipe.

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

On the other hand, the camellia oil extract of the present invention is a natural substance, which is harmless to the human body and has little toxicity and side effects, so it can be used safely even for long-term use.

The composition comprising the camellia oil extract of the present invention shows antibacterial activity against Trichomonas barnalis ( T. Vaginalis ) and anaerobic strain Gardnerella barnalis ( G. Vaginalis ), which cause female vaginitis, in feminine vagina even Lactobacillus know to maintain the slightly acidic environment necessary Ruth (L. acidophilus), Lactobacillus Creative spa tooth (L. crispatus), Lactobacillus joined Lee does not affect the growth of (L. gasseri) preventive efficacy of the female yeast infection It can be usefully used as a composition having a. In addition, the natural extract has little side effects on the human mucosa and can be safely used.

1 is a graph showing the growth of Camellia oil extract for three lactic acid bacteria.
Figure 2 shows the results of mucosal stimulation experiments with camellia oil extract.

Hereinafter, examples and the like will be described in detail to help understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be interpreted as being limited to the following examples. The 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 camellia oil extract

Camellia oil was used for the oil from Jeju camellia seeds, and the remaining oil was used. 2 L of purified water was added to 40 g of the above raw material, and hot water was extracted from a sterilizer. The supernatant was concentrated using a concentrator, and then lyophilized to powder. The powdered extract was dissolved in sterilized distilled water at a concentration of 100 mg/mL, and then used in an experiment after passing through a 0.2 μm pore size water-soluble injection filter.

Preparation Example 1: Experimental strain

Gardnerella barnacles ( G. vaginalis , ATCC14018) and Trichomonas barnacles ( T. vaginalis , ATCC PRA-95) were sold from the American Type Culture Collection, Lactobacillus acidopilus ( L. acidophilus , KCTC3164). , Lactobacillus crispatus ( L. crispatus , KCTC5054), Lactobacillus gasseri ( L. gasseri , KCTC3163) was used by pre-sale from the Korea Research Institute of Bioscience and Biotechnology.

Gardnerella barnacles ( G. vaginalis ) was anaerobic cultured at 37°C using an anaerobic gas pack (thermo scientific) in Brain Heart Infusion medium containing 10% FBS, and Trichomonas barnalis ( T. vaginalis ) was anaerobic cultured at 37°C using ATCC medium 361 containing 10% FBS. Three lactic acid bacteria were anaerobic cultured at 37°C in MRS culture/baby.

Example 1: Camellia Eucalyptus Extract Gardnerella barnalis ( Gardnerella vaginalis ) And Trichomonas overalls ( Trichomonas vaginalis ) To measure the impact on growth

In order to confirm whether the camellia oil extract of Preparation Example 1 has a growth inhibitory effect of a vaginitis-inducing strain, it was tested as follows. Before the experiment, the gardnerella pannalis cultured in a BHI agar containing 10% FBS was scraped into a loop and put in a BHI culture containing 10% FBS for static incubation under anaerobic conditions, and used in the experiment. The extracts were treated by concentration and inoculated so that the final number of bacteria was 1 X 10 ⁶ CFU/ml, followed by anaerobic culture for 24 hours. Trichomonas panagnalis was anaerobic cultured in ATCC medium 361 containing 10% FBS and used in the experiment. The extract was treated by concentration in a 96-well microplate and inoculated to have a final number of bacteria of 5 X 10 ⁴ cells/ml, followed by incubation for 24 hours. The untreated group was used as a negative control, and the strain that had been cultured was measured for absorbance at a wavelength of 600 nm to confirm the minimum inhibitory concentration. The results are shown in Table 1 below.

Strain name Minimum inhibitory concentration Gardnerella vaginalis 1 mg/ml Trichomonas vaginalis 0.5 mg/ml

As shown in Table 1 above, the camellia oil extract has a minimum inhibitory concentration of 1 mg/ml and 0.5 mg/ml, respectively, for the Gardnerella spp. And it was confirmed that it has a growth inhibitory effect on Trichomonas barnacles.

Example 2: Measurement of the effect of camellia oil extract on the growth of lactic acid bacteria

The following experiment was conducted to confirm whether the camellia oil extract has an effect on the growth of lactic acid bacteria, which are beneficial to humans. Lactobacillus acidophilus, Lactobacillus crispatus, and Lactobacillus gasseri) cultured in MRS agar were used for experiments by anaerobic culture in MRS culture. The extract was treated by concentration in a 96-well microplate and inoculated to have a final number of bacteria of 5 X 10 ⁶ CFU/ml, followed by incubation for 24 hours. After culturing, after confirming whether or not the strain was killed with the naked eye, the absorbance of each concentration was measured at a wavelength of 600 nm. The result of comparing the growth degree of the control group and the experimental group, which is the untreated group, is shown in FIG. 1.

1 is a graph showing the growth of Camellia oil extract for three lactic acid bacteria. As can be seen here, it was judged that there was no significant difference in absorbance between the treated and untreated Camellia oil extract groups, and thus it had little effect on the survival of lactic acid bacteria.

Example 3: Confirmation of mucosal stimulation by camellia oil extract

In order to confirm the irritation of the camellia oil extract on the mucous membrane, an HET-CAM test, an in-vivo replacement test, was performed. For the HET-CAM test, a modified brown Hy-line egg sold by Pulmuone was used, and cultured for 9 days at 37.5°C and 50% humidity. During the incubation period, the eggs were automatically rolled every hour, and on the 9th day, the eggs were removed from the incubator and used for testing after supplying moisture to the surface using 0.9% saline. The pores of the eggs were marked, and the inner shell was removed without destroying the inner membrane using a dental saw. In the HET-CAM analysis, concentrations of stimulation were determined based on the level of bleeding, dissolution, and coagulation caused by treatment with 0.1 N sodium hydroxide and 1% dodecyl sulfate sodium salt, which are known to cause stimulation, and camellia oil extract was 1 % Concentration. After 0.3 ml of the test substance was applied to the CAM, it was observed for 5 minutes to observe bleeding, melting, and coagulation reactions, and compared with when the reference substance was treated, scored using Equation 1 below, and the results are shown in FIG. 3. It is shown in.

In the response, 0 indicates no stimulus, 1 indicates weak stimulus, 2 indicates normal stimulus, and 3 indicates severe stimulation.

Figure 2 shows the results of mucosal stimulation experiments with camellia oil extract. As can be seen here, since the reaction was 0 under the conditions treated with the test group, the camellia oil extract was determined to be a safe substance without irritation to the mucous membrane.

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

Formulation Example 1: Female cleansing agent

1-1. Bubble type

ingredient weight% Camellia oil extract 1.00 Butylene Glycol 2.00 glycerin 1.00 Disodium LED 0.02 PAGE-60 Hydrogenated Castor Oil 1.00 Cocamidopropyl Betaine 4.00 Lauryl glucoside 6.00 Polysorbate 20 1.00 Sodium benzoate 0.40 Caprilic Glycol 0.20 Sodium lactate 0.50 Purified water 82.88 sum 100

1-2. Gel type

ingredient weight% Camellia oil extract 1.00 Butylene Glycol 2.00 glycerin 1.00 Disodium LED 0.02 PAGE-60 Hydrogenated Castor Oil 1.00 Cocamidopropyl Betaine 4.00 Lauryl glucoside 6.00 Polysorbate 20 1.00 Sodium benzoate 0.40 Caprilic Glycol 0.20 Sodium lactate 0.50 Hydroxyethyl cellulose 0.7 Purified water 82.18 sum 100

Claims (7)

A composition having antibacterial activity, characterized by comprising a camellia oil extract as an active ingredient. According to claim 1,
A composition characterized in that the activity is antibacterial activity against Gadnerella vaginalis and Trichomonas vaginalis . According to claim 1,
A composition characterized in that the camellia oil extract is contained in an amount of 0.0001 to 20% by weight relative to the total weight of the composition having antibacterial activity. A composition for improving or preventing female vaginitis comprising the composition having an antibacterial activity according to any one of claims 1 to 3 as an active ingredient. A female cleanser composition comprising the composition having an antibacterial activity according to any one of claims 1 to 3 as an active ingredient. The method of claim 5,
A female cleanser composition, wherein the female cleanser composition is a detergent, a pharmaceutical, a cosmetic, or a wipe. The method of claim 6,
The detergent cleaning agent is a female cleanser composition characterized in that it is a spray, solid, liquid, gel-type soap or foam-type cleaner.

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