KR20220146854A - A Cultured Gambir and Use thereof - Google Patents

Abstract

The present invention provides a catechu culture consisting of a culture obtained by culturing catechu with lactic acid bacteria and uses thereof. The present invention has the advantage of being effective for vaginal health, such as suppression of cervical cancer. The bacteria of the genus Lactobacillus can be at least one selected from Lactobacillus iners, Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus gasseri, Lactobacillus fermentum, and Lactobacillus rhamnosus.

Description

A Cultured Gambir and Use thereof

The present invention relates to a subgenetic drug culture and its use, and more particularly, to a subgonal drug culture effective for vaginal health, such as suppression of cervical cancer, and its use.

It is used as a dye and is also used as a herbal medicine. In the Korean Pharmacopoeia, it is described as a drug that is a dried aqueous extract obtained from the leaves and young branches of Uncaria gambir Roxburgh (Rubiaceae). See publication US2001/0012524 A1).

On the other hand, for women's health, it is necessary to develop a technology capable of responding to factors that threaten vaginal health, such as cervical cancer.

U.S. Patent Publication No. US2001/0012524 A1, (1st August 2001), Specification

One problem to be solved by the present invention is to provide an effective subchondral culture for vaginal health.

In addition, another problem to be solved by the present invention is to provide the use of a subchondral culture.

The problems of the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention provides a subchondral culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

The lactic acid bacteria may be bacteria of the genus Lactobacillus.

The Lactobacillus genus bacteria are Lactobacillus iners (Lactobacillus iners), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus jenseni (Lactobacillus jensenii), Lactobacillus gasseri (Lactobacillus gasseri), Lactobacillus permentum (Lactobacillus gasseri), Lactobacillus ), and Lactobacillus rhamnosus (Lactobacillus rhamnosus) may be at least one selected from.

It may be 2 to 6 parts by weight of the lactic acid bacteria with respect to about 100 parts by weight of the subchondral.

The culture may be a purified product that has undergone a lactic acid bacteria removal process after the culture.

The removal process may be a filtration process, and the purified product may be a filtrate.

The culture may be carried out at 37-42° C. for 24-48 hours.

The subchondral culture may be effective for vaginal health.

The subchondral culture may be for feminine hygiene, vaginal cleaning, or for treatment, suppression, improvement and/or prevention of cervical cancer.

In addition, the present invention provides the use of a feminine cleanser, a vaginal cleanser, and/or a use for the treatment, suppression, improvement and/or prevention of cervical cancer of the subchondral culture.

In addition, the present invention provides a feminine cleanser comprising a subchondral drug culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

The subchondral culture may be an active ingredient.

In addition, the present invention provides a vaginal cleansing agent comprising a subgonad culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

The subchondral culture may be an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the treatment or prevention of cervical cancer comprising, as an active ingredient, a subchondral drug culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

In addition, there is provided a cosmetic composition for improving or inhibiting cervical cancer, comprising as an active ingredient a subchondral drug culture consisting of a culture obtained by culturing subchondral drug with lactic acid bacteria.

In addition, the present invention provides a method for treating, suppressing, improving and/or preventing cervical cancer, comprising administering to a mammal, including a human, a subgonad culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

The administered subgenetic drug culture may be an effective amount of the subgenetic drug culture.

In addition, the present invention provides a use for the manufacture of a preparation for the treatment, suppression, improvement and/or prevention of cervical cancer of the subchondral drug culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria.

The present invention has the effect of being effective for vaginal health, such as suppression of cervical cancer.

1 is a graph showing the effect of the subchondral culture on the suppression of cervical cancer.
Figure 2 is a graph showing the effect of the extract of ahseonyak on the suppression of cervical cancer.
3 is a graph showing the effect of cisplatin on the suppression of cervical cancer.

Hereinafter, the advantages and features of the present invention, and a method of achieving them, will become apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, only these embodiments allow the disclosure of the present invention to be complete, and common knowledge in the technical field to which the present invention belongs It is provided only to fully inform the possessor of the scope of the invention, and the present invention is only defined by the scope of the claims.

Throughout the specification “and/or” includes each and every combination of one or more of the recited elements.

The terminology used herein is for the purpose of describing the embodiments and is not intended to limit the present invention. In this specification, the singular also includes the plural, unless specifically stated otherwise in the phrase. As used herein, “comprises” and/or “comprising” refers to the presence of one or more other components, steps, operations and/or elements mentioned. or addition is not excluded.

As used herein, 'treatment' means improvement or disappearance of a disease, pathological condition, or symptom. In addition, 'improvement' is included in 'treatment' and means that a pathological condition or symptom is improved. In addition, 'prevention' is meant to include preventing a pathological condition or symptom from worsening or preventing the transition to a disease. In addition, 'suppression' includes 'prevention', and includes preventing the occurrence of a pathological condition or symptom in addition to prevention.

In this case, inhibition or improvement may be applied to cosmetic use, and treatment or prevention may be applied to pharmaceutical use. That is, in the case of cosmetics, it may be a 'cosmetic composition for improvement or inhibition', and in the case of a drug, it may be a 'treatment or prophylaxis drug' or a 'therapeutic or prophylactic pharmaceutical composition'.

In addition, 'culture' is meant to include 'fermented product' indicating a fermentation product produced by a fermentation reaction.

One embodiment of the present invention is a subchondral drug culture consisting of a culture obtained by culturing subchondral drug with lactic acid bacteria. Such subchondral culture is effective for vaginal health. Specifically, it is effective for cervical cancer, and effective vaginal cleansing is also possible. As confirmed from the experimental examples, such an effect is difficult to be achieved with the subchondral drug alone, and it seems to be achieved by the complex effect of the cultured subchondral drug with the lactic acid bacteria.

Gambir (阿仙藥, Gambir) may be a ginseng extract. The chrysanthemum extract may be an extract (eg, a dried product of a water extract) obtained by extracting all or a part (eg, leaves and young branches) of a rhizome tree {Uncaria gambir Roxburgh (Rubiaceae)) with a solvent. In addition, ahseonyak extract is meant to include an extract called catechu, which is the same herbal name as ahseonyak. For example, the extract of Acacia cateche (legume) does not exclude extracts obtained by extracting all or part of Acacia cateche Willd (eg, heartwood) with a solvent (eg, dried water extract). The rhizome extract can be obtained by extracting the ahseonyak raw material with a hydrophilic solvent. The hydrophilic solvent may be at least one selected from water and a lower alcohol having 1 to 5 carbon atoms. The raw material of the subsea ginseng tree may be at least one selected from the group consisting of a chrysanthemum tree, a solvent extract of the rhododendron tree, Acacia catechu, and Acacia catechu. The solvent may be a hydrophilic solvent, and the hydrophilic solvent may be at least one selected from water and a lower alcohol having 1 to 5 carbon atoms. That is, the azalea extract may be an extract obtained by directly extracting all or part of the azalea tree and/or Acacia catechu with a solvent, or may be a re-extract obtained by extracting such an extract again with a solvent. For example, it may be a re-extraction obtained by re-extracting the dried aqueous extract obtained from the leaves and young branches of Uncaria gambir Roxburgh {Uncaria gambir Roxburgh (Rubiaceae)} described in the Korean Pharmacopoeia with a solvent. The azalea is preferably an extract obtained by extracting the dried aqueous extract obtained from the leaves and young branches of Uncaria gambir Roxburgh (Uncaria gambir Roxburgh) with one or more hydrophilic solvents selected from water and lower alcohols having 1 to 5 carbon atoms.

The extract includes a crude extract or a specific solvent-soluble extract (fraction), and such an extract may be in the form of a solution, a concentrate, or a dried product. The dried product may be a freeze-dried product or the like. When alcohol is used as an extraction solvent in such an extract, the alcohol may be evaporated by separately heating the extract or under reduced pressure in order to reduce the alcohol content in the composition. Specifically, alcohol may be removed by evaporating under heating and reduced pressure conditions using a rotary evaporator or by concentration using a reduced pressure concentrator. At this time, the alcohol content in the extract may preferably be 1.0% by weight or less.

The method for preparing such an extract is not limited thereto, and a commonly used extraction method may be applied. For example, hot water extraction method, ultrasonic extraction method, reflux cooling extraction method, etc. may be applied.

The lactic acid bacteria to be cultured together with the antibacterial drug may preferably be a bacterium of the genus Lactobacillus. The bacteria of the genus Lactobacillus are preferably Lactobacillus iners, Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus gasseri. Lactobacillus fermentum), and Lactobacillus rhamnosus may be at least one selected from the group consisting of (Lactobacillus rhamnosus). As such, the Lactobacillus genus bacteria exist as beneficial bacteria in the vagina, and thus, by culturing together with the antibacterial drug, it seems to have more effect on vaginal health. In addition, even if such lactic acid bacteria remain in the culture, it seems that it may be helpful to vaginal health.

Based on about 100 parts by weight of the subchondria, the lactic acid bacteria may be 2 to 6 parts by weight. In such a range, the culture proceeds effectively, and there is a fear that the culture is insufficient or unnecessary when it is less than or exceeding such a range.

The culture may be a purified product that has undergone a lactic acid bacteria removal process after culture. Since the presence of lactic acid bacteria is suppressed in such a purified product, it is possible to avoid unnecessary culture. The removal process may be, for example, a filtration process, and thus the purified product by filtration may be a filtrate. Filtration may be by means of a filter, filter paper, or the like.

The culture is not limited thereto as long as a culture, preferably a fermented product is formed, but preferably at 37 to 42° C., it may be carried out for 24 to 48 hours. In less than or more than such a range, there is a fear that the culture (specifically, fermentation by culture) is insufficient or excessive. Cultivation may be made in a medium, and the medium may be a lactic acid bacteria medium. The lactic acid bacteria medium is not limited as long as it is a medium in which the lactic acid bacteria can grow and preferably the fermentation reaction can proceed. The medium preferably contains a carbon source such as glucose. In addition, the medium may further include a nitrogen source (eg, yeast extract, etc.), amino acids, metals, and/or vitamins. Such a medium may be manufactured by a commercially available product or a well-known technique. Lactic acid bacteria grow best in weakly acidic conditions, but since the lactic acid bacteria itself lowers the pH and forms better growth conditions in the process of growth, neutral to weakly basic conditions are possible. Preferably, the pH of the medium may be 5 to 8. Oxygen conditions are not limited as long as the lactic acid bacteria can grow, but an oxygen partial pressure of 2 to 10% is preferable. At such oxygen partial pressure, lactic acid bacteria, which are preferably microaerobic bacteria, can grow better.

As an embodiment, the subchondrial culture is effective for vaginal health, as confirmed from experimental examples. Specifically, the subchondrial culture of an embodiment is capable of female cleanliness, vaginal washing, cervical cancer treatment, suppression, improvement and/or prevention, so it is possible for female cleanliness, vaginal washing, or cervical cancer treatment, suppression, improvement and/or It may be for prevention.

Therefore, the subchondral culture, which is an embodiment of the present invention, can be applied to a feminine cleanser or a vaginal cleanser, and can be used for medicinal and/or cosmetic purposes. For example, a feminine cleanser, a vaginal cleanser, a pharmaceutical composition, and/or a cosmetic composition according to an embodiment of the present invention includes a subchondral culture according to an embodiment of the present invention.

An embodiment of the feminine cleanser of the present invention includes a subchondral culture of an embodiment of the present invention. Such a subgenetic culture may be an active ingredient. Feminine cleanser may belong to cosmetics. Therefore, only the subgonad culture can be prepared and used in the same manner as the cosmetic preparation by adding the cosmetically acceptable additive to the subgonad culture or in the form of packaging in the container. Of course, the same contents as the cosmetic composition to be described later may be applied to the feminine cleanser.

In addition, the vaginal cleansing agent of an embodiment of the present invention includes a subgonad culture of an embodiment of the present invention. Such a subgenetic culture may be an active ingredient. Vaginal cleansers may belong to pharmaceuticals. Therefore, only the subchondrial culture can be prepared and used in the same way as a pharmaceutical formulation by adding a pharmaceutically acceptable additive to the subgonad culture or in the form of packaging in a container. Of course, the vaginal cleanser may have the same contents as the pharmaceutical composition to be described later.

In addition, the pharmaceutical composition according to an embodiment of the present invention may be a pharmaceutical composition for the treatment or prevention of cervical cancer, and includes the subchondrial culture according to an embodiment of the present invention as an active ingredient.

In addition, the cosmetic composition according to an embodiment of the present invention may be a cosmetic composition for improving or inhibiting cervical cancer, and includes the subchondrial culture according to an embodiment of the present invention as an active ingredient.

In addition, the method for treating, suppressing, improving and/or preventing cervical cancer according to an embodiment of the present invention may include administering to a mammal, including or not including a human, in need of administration of an active ingredient. The subgenetic culture to be administered may be an effective amount of the subgenetic culture.

In addition, the use for the preparation of a preparation for the treatment, improvement, inhibition, and/or prevention of cervical cancer, which is an embodiment of the present invention, is for the preparation of a preparation for the treatment, improvement, suppression, and/or prevention of cervical cancer of a subchondral culture. may be of use.

Unless otherwise stated, matters mentioned in uses, vaginal cleanser, feminine wash, pharmaceutical composition, cosmetic composition, and method apply the same, respectively, unless they contradict each other.

The active ingredient consisting of a subchondral culture is administered orally or parenterally, more preferably intravaginally, to mammals including humans. The pharmaceutical composition may be formulated by combining the active ingredient with a pharmaceutically acceptable carrier. For formulation, commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants, diluents, pH adjusters or excipients may be used. The disintegrant may be sodium starch glycolate, anhydrous calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the active ingredient, for example, microcrystalline cellulose, lactose, low-substituted hydroxycellulose, starch , calcium carbonate, sucrose, and/or gelatin may be added. In addition, a lubricant such as magnesium or talc may be used. Liquid formulations for oral administration include suspensions, internal solutions, emulsions, and syrups, and various additives such as wetting agents, sweetening agents, fragrances and/or preservatives in addition to commonly used simple diluents such as water and liquid paraffin. etc. may be included. Formulations for parenteral administration include injectable solutions, suspensions, emulsions, lyophilisates, nasal washes, and suppositories. Injectable solutions, suspensions, and emulsions can be prepared by mixing water, non-aqueous solvents or suspensions with active ingredients, and non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and ethyl oleate. It may be an injectable ester, such as As the base of the suppository, witepsol, macrogol, Tween 61, cacao butter, laurin fat, glycerol and/or gelatin may be used. When administered parenterally, it can be administered by subcutaneous injection, intravenous injection or intramuscular injection. Transdermal administration such as vaginal administration is also possible, and the external preparation for skin may be an external solution, cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, or a combination thereof. .

External preparations for skin are components commonly used in external preparations for skin such as cosmetics or pharmaceuticals, such as aqueous components, oily components, powder components, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, A pH adjuster, various skin nutrients, etc. can be suitably mix|blended as needed.

External preparations for skin include metal-blocking agents such as disodium edetate, sodium edetate hydrate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannins, bellapamil, licorice extract, Glably Dean, various herbal medicines, tocopherol acetate, glycyrrhizic acid, tranexamic acid and derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, trehalose, etc. saccharides, acids, and pH adjusters such as bases can also be appropriately blended.

The cosmetic composition may further include an additive acceptable for cosmetics in addition to the active ingredient.

Additives that can be added to the cosmetic composition may include components commonly used in cosmetic compositions, and include, for example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments and/or fragrances.

In addition, additives that can be added to the cosmetic composition include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, preservatives, bactericides, antioxidants, pH adjusters, alcohols, colorants, and/or fragrances. can be heard

The cosmetic composition may be prepared in any formulation conventionally prepared in the art, and for example, a solution, emulsion, cream, lotion, pack, foundation, lotion, cosmetic liquid, hair cosmetic, etc. may be mentioned.

When the formulation of the cosmetic composition is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. are used as carrier components. can be

When the formulation of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. propellants such as fluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene fatty acid esters of glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation of the cosmetic composition is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used.

Matters mentioned in the pharmaceutical composition or cosmetic composition according to an embodiment of the present invention are equally applied unless they contradict each other.

The active ingredient may be formulated by adding additives such as pharmaceutically or cosmetically acceptable carriers, excipients or diluents, and for information on formulation, refer to well-known literature related to formulation.

Accordingly, the pharmaceutical composition or cosmetic composition according to an embodiment of the present invention further comprises a pharmaceutically acceptable additive, or a cosmetically acceptable additive, and an active ingredient and a pharmaceutically acceptable additive, or a cosmetically acceptable additive. It may consist of additives.

The composition including the active ingredient, which is an embodiment of the present invention, preferably contains 0.001 to 99.99% by weight of the active ingredient, more preferably 0.001 to 80% by weight.

In addition, the active ingredient included in the composition according to an embodiment of the present invention or used in the method is 0.1 mg to 200,000 mg per day on an adult female basis (60 kg), for example, 0.1 mg to 1,000 mg, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or in doses of 10 mg to 25 mg. Administration may be administered once or divided into several times a day. However, the scope of the present invention is not limited by the dosage and number of administration.

Hereinafter, with reference to Examples, Experimental Examples, and Formulation Examples, an embodiment of the present invention will be described in more detail. For the reagents used in the experimental examples, the best commercially available reagents were used, and those purchased from Sigma, etc. were used.

<Experimental Example 1> Confirmation of cervical cancer inhibitory effect

1-1. Sample preparation

1-1-1. Example 1 (substance drug culture) preparation

Ahseonyak (represented in the Korean Pharmacopoeia) was purchased at Gyeongdong Market (located in Seoul, Korea). Methanol aqueous solution (70% w/v) was poured into the pulverized product of 10 g of the purchased phlegmon to a volume of 800 mL, and after 24 hours at room temperature, it was filtered through a filter to obtain a primary filtrate. Methanol aqueous solution (70% w/v) was poured into the residue remaining after the first filtration to a volume of 800 mL, and after 24 hours at room temperature, the second filtrate was obtained by second filtration with a filter. After combining the primary filtrate and the secondary filtrate, the mixture was concentrated under reduced pressure until the methanol layer was removed and a water layer remained. After the concentrate was cooled at -80°C for 24 hours, the frozen sample was freeze-dried (-30°C, vacuum degree 5 mtorr, 96 hours) to obtain a freeze-dried product (3.84 g). The freeze-dried product was used as a subchondral extract for culturing with lactic acid bacteria.

As for the lactic acid bacteria, L. acidophilus (KCCM 32820) was purchased from the Center for Biological Resources (KCTC) and used. The received L.acidophilus is diluted in distilled water, streaked on MRS medium (KCTC Media No. 178; Oxoid CM359; Difco 0881) agar plate, and pre-cultured for 24 hours. Take a single colony of the pre-cultured bacteria and incubate once more on an agar plate in MRS medium (KCTC Media No. 178; Oxoid CM359; Difco 0881) for 24 hours. Take one cultured single colony using platinum ear, inoculate it in 5ml liquid MRS medium (KCTC Media No. 178; Oxoid CM359; Difco 0881), and incubate for 24 hours. After the L. acidphilus cultured in the liquid medium was separated from the supernatant using a centrifuge, it was washed once more using RPMI medium {RPMI 1640 Medium (1X), Liquid}. The isolated bacteria were dissolved in RPMI medium to measure OD 600, and then diluted in RPMI medium to adjust OD 600 to 0.02. After that, the ahseonyak extract was added to make the concentration 600 μg/ml. After culturing the treated liquid medium for 24 hours (culture conditions: 37 degrees Celsius, 24 hours, 110 RPM shaking, culture in a zipper bag with a gas pack), the lactic acid bacteria were submerged with a centrifuge. Thereafter, the liquid was filtered using a 0.2 μm syringe filter. In this way, the supernatant from which the bacteria was filtered was prepared as a subgrade drug culture. The subchondrial culture was stored at -80°C and used for subsequent experiments.

1-1-2. Preparation of Reference Example 1 (Chinese ginseng extract)

A hyaluronic acid extract was prepared in the same manner as the azane medicine extract prepared in 1-1-1.

1-2. Experiment to confirm the effect of suppressing cervical cancer

The experiment was conducted using the Hela cell line (obtained from the Korea Cell Line Bank), which is a cervical cancer cell. Hela cell line 1.1x10 ⁵ cells/cm ² was dispensed in 7ml of RPMI medium containing 1% P/S (Penicillin/Streptomycin) in 10% FBS and 5% CO ₂ , and cultured in an incubator at 37° C. for 2 days. Cells were subcultured every 2 days. Hela cells were seeded in a 96-well plate at a concentration of 0.7x10 ⁵ /ml per well, and Hela cells were also seeded in each of two separate 96-well plates in the same manner, followed by stabilization at 37°C for 24 hours. Samples were injected into 3 wells at each concentration of 50ml each. Concentrations of each sample were 0, 100, 200, 300 ug/ml in the case of the hyaluronic acid extract, 0, 6.25, 12.5, 25, 50, 100, 200, 300 ug/ml in the case of cisplatin, 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, 100 uM. The lyophilisate extract sample is first dissolved in distilled water at a concentration of 100 mg/ml, and then diluted with RPMI medium {RPMI 1640 Medium (1X), Liquid} so that the concentration of the azalea extract is 100, 200, and 300 ug/ml, respectively. did. 0 ug/ml of the ahseonyak extract indicates that only RPMI medium was used as a sample. 0 ug/ml of the chalazion drug culture concentration indicates that the culture solution prepared in the same manner as in 1-1-1. For cisplatin samples, cisplatin was first dissolved in dimethyl sulfoxide (DMSO) at a concentration of 100 mg/ml, and then cisplatin concentrations of 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 uM were obtained with RPMI medium, respectively. It was diluted as much as possible and used. Cisplatin 0 uM indicates that only RPMI medium was used as a sample. Cisplatin is a known anticancer agent, and the cisplatin-treated group was prepared as a positive control group. After injecting the sample into each well, incubate for 48 hours at 37 degrees Celsius (5% CO ₂ ). After that, put the MTT reagent, block the light, and incubate for 4 hours at 37 ℃ (5% CO ₂ ). After suctioning the MTT reagent, dissolve it in 50ul of dimethyl sulfoxide (DMSO) to check cell viability at 540 nm absorbance. Cell viability is calculated by Equation 1 below.

[Equation 1]

Cell viability (%)= [(OD540 of sample) / (OD540 of control)]X100

As a control, the culture solution prepared in the same manner as in the preparation of the subchondral drug culture in 1-1-1.

The results are shown in FIGS. 1 to 3 .

Fig. 1 is a graph showing the effect of the subchondrial culture on the suppression of cervical cancer (* p<0.05: compared with the negative control group), Fig. 2 is a graph showing the effect of the subchondrial extract on the suppression of cervical cancer, and Fig. 3 is cis It is a graph showing the effect of platin on the suppression of cervical cancer. In each graph, the x-axis represents the concentration of the sample, and the y-axis represents the cell viability.

First, from FIG. 3, it can be seen that cisplatin, a known anticancer agent, inhibits cervical cancer cells in a concentration-dependent manner, and from these results, it can be seen that the experiment was performed normally.

From FIG. 2, it can be seen that the ahseonyak extract itself does not have a great effect on suppressing cervical cancer cells.

However, it appears that the subchondrial culture inhibits cervical cancer cells in a concentration-dependent manner (see FIG. 1). In addition, it can be seen that in the case of a sample treated with only lactic acid bacteria culture (0ug/ml in FIG. 1), it can be seen that the cervical cancer cell inhibitory effect is not shown.

From these results, an effect effective on vaginal health, such as suppression of cervical cancer cells, is difficult to be achieved only with a choline drug or lactic acid bacteria, but it seems to be achieved by a complex effect by a culture obtained by culturing the opiates drug with lactic acid bacteria.

From these results, it can be seen that the subchondral culture, which is an embodiment of the present invention, effectively inhibits cervical cancer. Therefore, it can be seen that the present invention is effective for vaginal health and can be applied for the treatment, suppression, improvement and/or prevention of cervical cancer.

<Experimental Example 2> Human body application experiment

2-1. Preparation of subchondral culture

In the same manner as in 1-1-1., a subchondral culture was prepared.

2-2. usability evaluation

For each of the subchondrial culture and distilled water prepared in 2-1., 10 women (20s to 40s) living in Gyeonggi-do and Seoul, Korea were selected and usability evaluation was conducted during vaginal cleansing. Sample irritation, sample odor, odor after use, vaginal impurities, vaginal freshness, vaginal irritation, and vaginal itchiness were evaluated. After establishing stable judgment criteria repeatedly for each item, evaluation was carried out using the 5-point scoring method. The closer to 5 points, the greater the satisfaction.

The results are shown in Table 1. The data in Table 1 shows the average value and standard deviation for each item of each sample.

Subsidiary culture Distilled water Irritation of the sample 4.7±0.48 4.4±0.52 sample odor 4.5±0.53 4.6±0.52 smell after use 4.3±0.67 3.9±0.74 vaginal impurities 3.9±0.57 2.9±0.57 vaginal freshness 4.0±0.47 3.9±0.74 vaginal stimulation 4.2±0.79 4.6±0.52 vaginal itching 3.8±0.79 1.6±0.70

As shown in Table 1, it can be seen that, in the case of the subgonad culture, the satisfaction is very high in terms of odor, vaginal itchiness, and vaginal impurities after use in the vagina. From these results, it can be seen that the subgonad culture according to an embodiment of the present invention is very effective in terms of vaginal cleaning or cleanliness. Therefore, the subchondrial culture of the present invention is effective for vaginal health, and can be applied as a vaginal cleanser and/or a feminine cleanser.

From the above experimental results, it can be seen that the subchondrial culture according to an embodiment of the present invention is effective for vaginal health, and specifically, it is effective for female cleanliness, vaginal cleansing, treatment, suppression, improvement and/or prevention of cervical cancer.

Therefore, it can be seen that the subchondral culture, which is an embodiment of the present invention, can be applied as a feminine cleanser, a vaginal cleanser, and can be applied as a medicine/cosmetics for the treatment, suppression, improvement and/or prevention of cervical cancer.

<Preparation Example 1> Preparation of pharmaceutical composition

After filtration with a cartridge filter (0.5um) the subgonad culture prepared in the same way as in Experimental Example 1-1-1., Natrotics (golden extract, magnolia skin extract, propolis extract, camellia leaf extract, Nahanbaek extract , German chamomile extract, and willow bark extract-containing mixture), and menthol were added, and the mixture was filtered again with a cartridge filter (10um). Thereafter, after adjusting the remaining amount by adding purified water, instantaneous sterilization is performed, and 200 ml of each is injected into a sterile container to prepare an external solution. Subsidiary culture, natrotics, and menthol should be 80% by weight, 2% by weight, and 1% by weight of the total weight of the external solution, excluding the weight of the container, respectively.

<Preparation Example 2> Preparation of cosmetic composition

A mixture of natrotix, glycerin, vitamin E, vitamin C1, and tea tree oil was added and mixed after filtering the subchondral culture prepared in the same manner as in Experimental Example 1-1-1 with a cartridge filter (0.5um). is filtered again with a cartridge filter (10um). Thereafter, after adjusting the remaining amount by adding purified water, instantaneous sterilization is performed, and 200 ml of each is injected into a sterile container to prepare an external solution. Subsidiary culture, natrotics, glycerin, vitamin E, vitamin C1, and tea tree oil are 50 wt%, 2 wt%, 5 wt%, 0.05 wt%, 0.05 wt%, 2 Let it be the content in weight %.

<Preparation Example 3> Preparation of a vaginal cleaner

After filtering the subchondral culture prepared in the same manner as in Experimental Example 1-1-1. with a cartridge filter (0.5um), natrotics and menthol were added, and the mixture was filtered again with a cartridge filter (10um). . Thereafter, after adjusting the remaining amount by adding purified water, instantaneous sterilization is performed, and 200 ml of each is injected into a sterile container to prepare a vaginal cleanser. Subsidiary culture, natrotics, and menthol should be 80% by weight, 2% by weight, and 1% by weight of the total weight of the vaginal cleanser excluding the weight of the container, respectively.

<Preparation Example 4> Manufacture of feminine cleanser

After filtration with a cartridge filter (0.5 μm) the subchondrial culture prepared in the same way as in Experimental Example 1-1-1., natrotics, menthol, and yucca extract (a natural surfactant obtained from yucca root) were added and mixed The mixture is filtered again through a cartridge filter (10um). Thereafter, after adjusting the remaining amount by adding purified water, instantaneous sterilization is performed, and 200 ml of each is injected into a sterile container to prepare a feminine wash. Subsidiary culture, natrotics, menthol, and yucca extracts should each be 50 wt%, 2 wt%, 1 wt%, or 2 wt% of the total weight of the feminine cleanser excluding the container weight.

Claims (7)

A subgenetic drug culture consisting of a culture obtained by culturing subchondral drug with lactic acid bacteria. According to claim 1, wherein the lactic acid bacteria is a bacterium of the genus Lactobacillus subchondrial culture. The subgenetic drug culture according to claim 1, which is effective for vaginal health. A feminine cleanser comprising a subchondral drug culture consisting of a culture obtained by culturing subchondral drug with lactic acid bacteria. A vaginal cleaning agent comprising a subgonad culture consisting of a culture obtained by culturing the subchondral drug with lactic acid bacteria. A pharmaceutical composition for the treatment or prevention of cervical cancer, comprising, as an active ingredient, a subchondral drug culture consisting of a culture obtained by culturing subchondral drug with lactic acid bacteria. A cosmetic composition for improving or inhibiting cervical cancer, comprising, as an active ingredient, a culture of subchondral drug obtained by culturing subchondral drug with lactic acid bacteria.

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