KR20210036213A - Antifungal Composition Comprising Rose Extract and Moutan Radicis Cortex Extract Having Synergistic Effect, and Composition for Preventing or Treating Candidasis - Google Patents

Abstract

The present invention relates to an antifungal composition and a composition for preventing or treating candidiasis which contain a rose extract and a Moutan radicis cortex extract having a synergistic effect, wherein the antifungal composition and the composition have excellent antifungal activity on Candida albicans, so that it is possible to be usefully utilized for the prevention, alleviation or treatment of various diseases caused by Candida albicans.

Description

Antifungal Composition Comprising Rose Extract and Moutan Radicis Cortex Extract Having Synergistic Effect, and Composition for Preventing or Treating Candidasis}

The present invention relates to an antifungal composition comprising a rose extract and a mokdanpi extract having a synergistic effect, and a composition for preventing or treating candidiasis.

Cutaneous mycosis, which is the most common form of skin disease in everyday life, can be transferred to other people by dead skin cells that have fallen from infected skin in places where several people are gathered, such as swimming pools, bathhouses, military units, and homes. It can be caused by diseases such as back or poor hygiene. If proper treatment is not done, it may be transferred to other parts of the body or cause deformity of nails and toenails. Most of the mycosis, once infected, is not treated well and progresses chronically, but it is recognized as a disease of not high severity. However, in the case of patients with reduced immunity, infection and mortality rates caused by opportunistic fungi such as Candida, Aspergillus and Cryptococcus are steadily increasing worldwide.

Among the various fungi, Candida albicans mainly reproduces in epithelial cells in the human body. Candida albicans exists in normal skin, mucous membranes, feces, sputum, urine, etc., and is harmless to the human body, but when antibiotics or adrenal cortical hormones are used, or when the body's immunity is reduced, it proliferates and causes disease. It can also cause local mucositis such as stomatitis and vaginitis, or can lead to systemic infections. Types of candidiasis caused by Candida albicans include mucosal candidiasis in which the mouth, tongue, vulva, and vagina are sore and painful, and skin candidiasis in which redness in the area of the anus or between the fingers, between the wrinkles of the skin or between the fingers, etc. , Diarrhea, abdominal pain, jaundice, and digestive tract candidiasis, sepsis, and the like, and systemic candidiasis accompanied by meningitis or endocarditis.

Currently, as antifungal substances to inhibit the growth of fungi, various types of antifungal agents such as Azole-based, Polyene-based and Allyamines-based have been developed. However, these antifungal agents exhibit various kinds of side effects including hepatotoxicity, and in the case of Azole-based drugs, which are most commonly used, the incidence of resistance rapidly increases. Accordingly, there is an increasing need for new antifungal substitutes having excellent antifungal effects, low side effects and low toxicity.

Accordingly, the present inventors completed the present invention by conducting a study to develop an antifungal composition using a safe natural plant extract.

Korean Patent Registration No. 10-1398426

One object of the present invention is to provide an antifungal composition comprising a rose extract and a mokdanpi extract.

Another object of the present invention is to provide a composition for feminine cleansing comprising the composition.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating candidiasis comprising a rose extract and a mokdanpi extract.

One aspect of the present invention provides an antifungal composition comprising a rose extract and a mokdanpi extract.

Roses are sweet in taste, warm in nature, non-poisonous, and have the effect of promoting blood circulation and reducing swelling. In addition, rose flowers are known to exhibit a tonic effect and blood pressure control effect by containing various ingredients such as various vitamins, organic acids, tannins, etc., as they have a very strong and clear scent, mild, and excellent aroma. However, no known antifungal efficacy is known.

Mokdanpi is the root bark of Paeonia suffruticosa Andrews belonging to the Paeoniaceae family. Mokdanpi extract inhibits the release of IL-8 and MCP-1, a kind of chemokine, in vitro, and it has been reported to inhibit the expression of iNOS and COX-2, which are inflammatory mediators. However, no known antifungal efficacy is known.

The antifungal composition comprising the rose extract and the mokdanpi extract of the present invention has excellent antifungal activity against Candida albicans, and can be usefully used in the prevention, improvement or treatment of various diseases caused by Candida albicans. The antifungal composition of the present invention may be usefully utilized as a composition for feminine cleansing and a pharmaceutical composition for the prevention or treatment of candidiasis.

The extract contained in the composition according to the present invention can be obtained as follows. Wash the rose or mock dandelion with water to remove foreign substances, and then dry it in the shade, and add an appropriate amount of solvent to the rose or mockup so that it is completely immersed.At this time, the rose or mockskin should be used as it is dried or pulverized to be used in the form of powder. I can. Rose or bark can be extracted using a conventional extraction solvent, and preferably, (a) an anhydrous or water-containing lower alcohol having 1 to 4 carbon atoms (e.g. methanol, ethanol, propanol, butanol, normal-propanol, iso- Propanol and normal-butanol), (b) a mixed solvent of the lower alcohol and water, (c) acetone, (d) ethyl acetate, (e) chloroform, (f) 1,3-butylene glycol, (g ) Hexane, (h) diethyl ether, (i) butyl acetate (j) chloroform-methanol, or (k) distilled water. Can be impregnated or heated at room temperature during extraction

According to one embodiment of the present invention, the extract may be extracted with one or more solvents selected from the group consisting of alcohols having 1 to 4 carbon atoms and a mixed solvent thereof.

According to one embodiment, the fungus may be a Candida albicans (C andida albicans).

Candida albicans is a part of the normal flora and is the causative agent of candidiasis, also known as moniliosis, which is an abnormal fungus such as yeast that clusters on the mucous membrane surface of the oral, genital and gastrointestinal tract of humans. Candida albicans is a type of fungus (fungus) that mainly reproduces in epithelial cells in the human body.

According to an embodiment of the present invention, the concentration ratio by volume of the rose extract and the mokdanpi extract may be 0.2 to 0.275: 0.625 to 0.65.

The combination of the rose extract and the mokdanpi extract contained in the composition of the present invention exhibits a synergistic effect on the growth inhibition of Candida albicans. In the combination of 0.2 to 0.275% (v/v) concentration of rose extract and 0.625 to 0.65% (v/v) concentration of Mokdan bark extract, synergistic effect appeared on the growth inhibition of Candida albicans, and increased inhibition of the growth of Candida albicans. The combination of the rose extract and the mokdanpi extract showing the effect is preferably a concentration ratio of 0.2 to 0.225% (v/v): 0.625 to 0.65% (v/v), and 0.2% (v/v): 0.625% (v/ It is most preferable that it is the concentration ratio of v).

Another aspect of the present invention provides a composition for a feminine cleansing agent comprising the composition.

The rose extract and mokdanpi extract of the present invention has excellent antifungal activity against Candida albicans, so it can be usefully used as a composition for feminine cleansing to prevent or improve female genitourinary diseases such as vaginitis caused by Candida albicans. have.

The term "feminine cleanser" used in the present invention refers to a dedicated cleanser used around and around the vulva of a woman for the purpose of inhibiting the proliferation of harmful bacteria in the vagina, reducing odor and/or maintaining an appropriate pH in the vagina.

The composition for feminine cleansing agent of the present invention may contain excipients, disintegrants and/or lubricants acceptable for feminine cleansing agents. Excipients, disintegrants and lubricants included in the composition for feminine cleansing agent of the present invention are commonly used in the manufacture of feminine cleansing agents, and the excipients are prosolve, starch, microcrystalline cellulose, mannitol, lactose, low-substituted hydroxypropyl cellulose And/or sucralose, specifically, Prosolve (Prosolve SMCC90), starch (Starch 1500), microcrystalline cellulose (microcrystalline cellulose PH102) and/or mannitol (mannitol 160C), and the disintegrant may be cross Povidone, sodium starch gluconate, croscarmellose sodium, polysorbate and/or sodium lyryl sulfate may be, specifically, crospovidone (crospovidone CL-SF), and the lubricant is magnesium stearate, hard It may be silicic anhydride, talc, and/or glyceryl behenate, and specifically, magnesium stararate, but is not limited thereto.

The composition for feminine cleansing of the present invention is for the prevention or improvement of diseases related to the vagina, and diseases related to the vagina may be vaginosis, vaginal dryness and/or vaginitis, but are not limited thereto.

According to one embodiment of the present invention, the composition may be any one formulation selected from the group consisting of a solution, a gel, a cleaning composition, and a tablet for vaginal insertion.

The present invention is a composition for a feminine cleanser, the formulation may be a formulation for parenteral administration, for example, a solution, a gel, a cleaning composition, a tablet for vaginal insertion, a suppository form, a cream, an ointment, a dressing It may be a liquid formulation such as a solution, a spray, and other coating agents, such as a solution type, a suspension type, an emulsion type, and a sterilized aqueous solution, a non-aqueous solution, a suspension, an emulsion, a freeze-dried preparation, a suppository, a cream, an ointment, It may be a jelly, a foam, a detergent, or a vaginal insert, and preferably a liquid, a gel, a cleaning composition, an external preparation for the skin such as a tablet for vaginal insertion.

The formulation can be prepared, for example, by adding a solubility aid, an emulsifier, a buffer for pH adjustment, and the like to sterile water. As the non-aqueous solvent or suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and/or ethyl oleate may be used. On the other hand, the suppository formulation may be prepared by adding, for example, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycero gelatin, etc., and a carrier for formulating it may be prepared. As the carrier, a binder, a lubricant, a suspending agent, a solubilizing agent, a buffering agent, a preservative, a lubricant, an isotonic agent, a stabilizer, a dispersant, a suspending agent, a colorant, a fragrance, and the like may be included as the carrier.

The preferred dosage of the composition for feminine cleansing of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art.

The dosage of the composition for feminine cleansing of the present invention may be appropriately selected by those skilled in the art in consideration of the administration method, the age, sex and weight of the user, and the severity of the disease. For example, the dosage of the composition for feminine cleansing of the present invention may be 0.001 to 1000 mg/kg based on an adult. It may be administered in divided doses once or several times a day, but is not limited thereto.

The composition for feminine cleansing of the present invention may be administered together with a substance exhibiting prophylactic or therapeutic activity for at least one female genitourinary system-related disease.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating candidiasis comprising a rose extract and a mokdanpi extract.

In the pharmaceutical composition for preventing or treating candidiasis of the present invention, portions overlapping with the above description may have the same meaning as described above.

As used herein, the term "candidasis" refers to various infectious diseases caused by Candida albicans. Candidiasis is divided into skin mucous candidiasis and deep candidiasis depending on the affected site. Skin mucous candidiasis includes thrush, esophageal candidiasis, and vaginal candidiasis. These include spondylitis and hepatosplenic candidiasis.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in the manufacture of drugs, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin. , Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. It is not limited. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (22th ed., 2013).

The pharmaceutical composition according to an embodiment of the present invention may be administered together with a substance exhibiting activity in the prevention or treatment of one or more candidiasis.

In addition, the pharmaceutical composition according to an embodiment of the present invention may be used alone for the treatment of candidiasis, or in combination with procedures, hormone treatments, drug treatments, and/or methods of using a biological response modifier.

The pharmaceutical composition of the present invention may contain various bases and/or additives that are necessary and appropriate for the formulation of the formulation, and within a range that does not degrade the effect, nonionic surfactants, silicone polymers, extender pigments, fragrances, preservatives , Disinfectant, oxidation stabilizer, organic solvent, ionic or nonionic thickener, softening agent, antioxidant, free radical destroying agent, opacifying agent, stabilizer, emollient, silicone, α-hydroxy acid, antifoaming agent, moisturizing agent , Vitamins, insect repellents, fragrances, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basification or acidifying agents, or coloring agents, and other known compounds.

The appropriate dosage of the pharmaceutical composition of the present invention is formulated in various ways depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity. Can be. The dosage of the pharmaceutical composition of the present invention may be 0.001 to 1000 mg/kg on an adult basis.

The pharmaceutical composition of the present invention can be administered parenterally.

The pharmaceutical composition of the present invention can be administered in various dosage forms when administered parenterally. Solid preparations include tablets, pills, powders, granules, capsules, etc., and liquid preparations include suspensions, liquid solutions, emulsions, and syrups. In addition to water, liquid, and paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, fragrances, and preservatives may be included. Specifically, formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions and freeze-dried formulations. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. In addition, calcium or vitamin D3 may be added to enhance the efficacy of the therapeutic agent.

Such compositions may be presented in unit-dose (single) or multi-dose (several batches) containers, e.g., sealed ampoules and vials, and immediately prior to use, in a sterile liquid carrier, e.g., water for injection. It can be stored under freeze-drying conditions requiring only the addition of. Ready-to-use preparations and suspensions can be prepared from sterile powders, granules and tablets.

According to an embodiment of the present invention, the concentration ratio by volume of the rose extract and the mokdanpi extract may be 0.2 to 0.275: 0.625 to 0.65.

According to an antifungal composition comprising a rose extract and a mokdanpi extract having a synergistic effect, and a composition for preventing or treating candidiasis, because it has excellent antifungal activity against Candida albicans, the prevention and improvement of various diseases caused by Candida albicans Or it can be usefully used for treatment.

1 is a graph showing the growth inhibitory effect on Candida albicans according to the concentration of rose extract.
Figure 2 is a graph showing the growth inhibitory effect on Candida albicans according to the concentration of mokdanpi extract.
Figure 3 is a diagram showing a treatment method of rose extract and mokdanpi extract.
Figure 4 is a diagram showing the growth inhibition rate (GI) of Candida albicans compared to the control according to the mixed treatment of rose extract and mokdanpi extract.
FIG. 5 is a diagram showing the calculated GI value obtained by mixing and treating the rose extract and the mokdanpi extract, divided by the calculated GI value treated with only the rose extract.
FIG. 6 is a diagram showing a calculated value obtained by dividing a calculated GI value obtained by mixing and treating a rose extract and a mokdanpi extract by the calculated GI value treated with only a mokdanpi extract.

Hereinafter, the present invention will be described in more detail through one or more examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1. Preparation of rose extract and mokdanpi extract

40 liters of water purified with a reverse osmosis filter was added to the extractor tank, crushed rose or crushed wood bark was put in a non-woven fabric and immersed in the tank, followed by extraction at 100°C for 3 hours and 30 minutes, after extraction, immersion for 12 hours, and natural cooling Then, it was filtered at 40 ℃ to prepare a rose extract and a mokdanpi extract, respectively.

Example 2. Preparation of strain and culture medium

In order to measure the antifungal effect, Candida albicans (C andida albicans , hereinafter C. albicans ) was purchased from KCTC 7965 from the Center for Biological Resources (Korean Collection for Type Cultures, Korea). The composition of the medium used in the experiment is shown in Table 1.

badge Composition content SDA Sabouraud Broth
(BD Difco, catalog number: 238230) 30.0g Agar 15.0g Distilled water Replenish the total volume to 1,000 ml RPMI 1640 RPMI-1640 Medium
(Merck KGaA, product number: R6504) 10.4g MOPS [3-(N-morpholino)propanesulfonic acid] 34.53g 1M Sodium hydroxide Use an appropriate amount with pH 7.0 Distilled water Replenish the total volume to 1,000 ml

SDA medium was used to make C. albicans stored at -80°C in an active growing state, RPMI 1640 medium was used to cultivate C. albicans in an active growing state, and the finished medium was stored at 4°C. Was used.

For use in the evaluation of growth inhibition, C. albicans KCTC 7965 strain stock was streaked on an SDA agar plate and cultured at 35°C for 1 day. A single cluster cultured on a new SDA agar plate was taken, streaked again, and incubated at 35°C for 1 day. Thereafter, a single cluster was taken, suspended in 0.145 mol/L saline, and vortexed for 15 seconds. Measure the absorbance of the suspension at 530nm using an Optizen 2120uv spectrophotometer (Mecasys Co., Korea) to confirm that the cell density becomes a McFarland standard of 0.5, and add saline solution to make 0.5 A suspension of C. albicans was prepared by adjusting the turbidity of McFarland.

Example 3. C. albicans Confirmation of the growth inhibitory effect of rose extract and Mokdan bark extract against and by mixing rose extract and Mokdan bark extract C. albicans The synergistic antibacterial effect on the

3-1. Dispensing of rose extract and mokdan bark extract to confirm the growth inhibitory effect and synergistic effect of the extract

In order to measure the antifungal effect on C. albicans , the CLSI M27-A2 test method was used. A Checkerboard assay was applied to confirm the synergistic antibacterial effect according to the concentration of the two extracts.

Specifically, the C. albicans suspension having a Macfaland turbidity of 0.5 prepared in Example 2 was diluted 50-fold with RPMI 1640 medium, diluted 20-fold and dispensed into microplates by 100 µl each, and then the extract dilution (extract was When mixed with 100 µl of a solution diluted with RPMI 1640 medium), the concentration of C. albicans was 1×10 ³ to 5×10 ³ CFU/ml.

On the other hand, the rose extract prepared in Example 1-1 was diluted with RPMI 1640 medium to prepare a stock having a concentration of 10% (v/v), from this stock 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, Four-fold concentration stocks were prepared to be 1.1% and 1.2%, respectively. These stocks were dispensed from low concentration to rows 3 to 10 of the microplate each 50 µl, and then mixed with 50 µl of medium or mokdanskin extract dilution and 100 µl of C. albicans inoculum, the final amount of rose extract contained in 200 µl. The treatment concentration was set to 0.15%, 0.175%, 0.2%, 0.225%, 0.25%, 0.275%, and 0.3%.

Similarly, the Mokdanpi extract prepared in Example 1-1 was diluted with RPMI 1640 medium to prepare a stock having a concentration of 10% (v/v), from this stock to 2.2%, 2.3%, 2.4%, 2.5%, 2.6%. Each four-fold concentration stock was prepared. From low concentration, 50 µl of each of these stocks was dispensed into rows C to F of the microplate, and then mixed with 50 µl of medium or rose extract dilution and 100 µl of C. albicans inoculum, The final treatment concentration was set to 0.55%, 0.575%, 0.6%, 0.625%, 0.65%.

In order to evaluate the growth inhibitory effect of the bacteria by concentration of rose extract and mokdanpi extract only, columns 3-10 of rows C-G and B of column 2 do not mix rose extract and mokdanpi extract, 50 µl of extract dilution, and RPMI. 50 µl of 1640 medium was dispensed, and 100 µl of C. albicans inoculum was added and cultured (Figs. 1 and 2). The control group was dispensed with 100 µl of RPMI 1640 medium instead of the extract dilution (FIG. 3).

After dispensing the rose extract dilution, Mokdanskin extract dilution, and C. albicans inoculum into a microplate, the microplate was incubated at 35°C for 48 hours, and then the sinking bacteria were suspended by pipetting, and a Microplate Reader (BioTek Microplate Reader) ) by measuring the absorbance at 530nm it was calculated cell density (cell density) of C. albicans, through the cell density value was evaluated the proliferation or growth results of C. albicans.

3-2. Confirmation of conditions of rose extract and mokdan bark extract showing synergistic effect

The GI (Growth inhibition of the concentration) value was calculated by calculating the cell density value of C. albicans measured in Example 3-1 by the following equation. GI _A is the GI value when only the rose extract is treated, GI _B is the GI value when only the mokdanskin extract is treated, and the GI _combination is the value when the rose extract and the mokdanskin extract are mixed and treated.

GI _A (%) = (OD _control -OD _A )/OD _control ×100

GI _B (%) = (OD _control -ODB)/OD _control ×100

GI _combination (%) = (OD _control -OD _combination )/OD _control ×100

Based on the calculated GI _A , GI _B, and GI _combination values, the synergistic effect of rose extract and mokdan bark extract against C. albicans was expressed as'synergistic effect, partial synergistic effect, no effect' using the following equation. .

2 <GI _combination / GI _A and GI _combination / GI _B : synergistic effect

1.5 <GI _combination / GI _A or GI _combination / GI _B ≤ 2: partial synergistic effect

GI _combination /GI _A or GI _combination /GI _B ≤ 1.5: No effect

The calculated GI value (FIG. 4) when the rose extract and the mokdanpi extract were mixed and treated was divided by the calculated GI value treated with only the rose extract (FIG. 5) and the calculated GI value processed only the mokdanpi extract (FIG. If all of 6) are greater than 2, it is judged that there is a synergistic effect. When the two values exceeded 1.5 and were 2 or less, it was judged that there was a partial synergistic effect, and when either of these two values was 1.5 or less, it was judged that there was no synergistic effect.

_{As a result, both GI combination} / GI _A value and GI _combination / GI _B value were more than 2 in the section in which 0.2 to 0.225% (v/v) of rose extract and 0.625 to 0.65% (v/v) of mokdanpi extract were mixed. , In the antifungal effect on C. albicans , it was confirmed that rose extract and mokdanpi extract exhibit a synergistic effect within the corresponding range. On the other hand, in the section in which 0.25 to 0.275% (v/v) of rose extract and 0.625 to 0.65% (v/v) of mokdanpi extract were mixed, both GI _combination / GI _A value and GI _combination / GI _B value were more than 1.5 and 2 or less. As a result, in the antifungal effect against C. albicans , it was confirmed that rose extract and mokdanskin extract exhibited a partial synergistic effect within the corresponding range.

So far, the present invention has been looked at focusing on the examples. Those of ordinary skill in the art to which the present invention pertains will be able to understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative point of view rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (7)

Antifungal composition comprising rose extract and mokdanpi extract.
The method of claim 1, wherein the fungus is an antifungal composition to the Candida albicans (C andida albicans)
The antifungal composition of claim 1, wherein the concentration ratio by volume of the rose extract and the mokdanpi extract is 0.2 to 0.275: 0.625 to 0.65.
A composition for feminine cleansing comprising the composition of claim 1.
The composition for feminine cleansing according to claim 4, wherein the composition is any one formulation selected from the group consisting of a solution, a gel, a cleaning composition, and a tablet for vaginal insertion.
A pharmaceutical composition for preventing or treating candidiasis comprising a rose extract and a mokdanpi extract.
The pharmaceutical composition for preventing or treating candidiasis according to claim 6, wherein the concentration ratio by volume of the rose extract and the mokdanpi extract is 0.2 to 0.275: 0.625 to 0.65.

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