KR20140116788A - Composition for protecting and treating vaginosis disease comprising bee venom effective for treatment of vaginitis - Google Patents
Composition for protecting and treating vaginosis disease comprising bee venom effective for treatment of vaginitis Download PDFInfo
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Abstract
The present invention relates to a composition for prevention and treatment of vaginitis comprising bee venom. More specifically, the present invention relates to a composition for prevention and treatment of vaginitis, It has excellent antimicrobial activity against fungi and general bacteria and is suitable for use in the prevention and treatment of vaginitis. In addition, it is excellent in formulations that can be manufactured in various formulations, and it is possible to commercialize products according to the taste and convenience of patients.
Description
The present invention relates to a composition for preventing and treating vaginitis including bee venom.
Vaginitis is the most common gynecological condition with vaginal infection or inflammation characterized by symptoms such as vaginal discharge, odor, burning sensation, bulimia, dyspareunia, and dysuria. More than 90% of the vaginitis in women of childbearing age are bacterial vaginitis, candidiasis, and trichomoniasis. Bacterial vaginitis is the most common vaginitis occurring at the age of childbirth, accounting for 40 ~ 50%, followed by Candida infections 20 ~ 25%, Trichomonas vaginitis 15 ~ 20% and noninfectious vaginitis. There are many kinds of normal bacteria in the vagina and lactobacillus , which is the aerobic bacterium, is the most common. The bacterium secretes hydrogen peroxide, which keeps the vagina acidic, maintains balance among organisms, and is resistant to germs. In situations where the resistance group changes, the chance of infection increases.
In the treatment of vaginitis, bacterial vaginosis uses drugs such as metronidazole and clindamycin, which are injected into the vagina and orally administered. Medication should be given during pregnancy, but metronidazole can be harmful to the fetus, so treatment should be avoided in the early stages of pregnancy and treatment should be started in the middle of pregnancy. Another method is oral administration of clindamycin or the like.
Candida albicans is an azole-type antifungal agent used in topical therapy. Examples of antifungal agents include butoconazole, clotrimazole, miconazole, nystatin, , Ticonazole and terconazole. Examples of antifungal agents used in oral therapy include fluconazole, itraconazole, and ketoconazole.
In the case of trichomoniasis vaginitis, metronidazole is administered orally. In the early pregnancy, metronidazole treatment is prohibited and treatment is possible from the middle of pregnancy. In addition, when treatment fails due to recurrence, antibiotic susceptibility test for metronidazole should be performed [Non-Patent Document 1].
Metronidazole and clindamycin, which are used in the treatment of vaginosis, have side effects of antibiotics and difficulty in treatment by resistant bacteria due to long-term treatment. Furthermore, since it is possible to adversely affect the fetus in the early pregnancy, And the like.
The present inventors have conducted studies to solve the above-mentioned problems. As a result, the present inventors have found that the use of natural products such as bee venom can reduce side effects on the human body, The present invention has been completed by developing a composition.
Accordingly, it is an object of the present invention to provide a composition for preventing and treating vaginitis comprising bee venom as an active ingredient.
As means for solving the above problems, the present invention provides a composition for preventing and treating vaginitis comprising bee venom as an active ingredient.
As another means for solving the above problems, the present invention provides a vaginal detergent, gel, suppository or aerosol preparation containing bee venom as an active ingredient.
The bee venom, which is an active ingredient used in the present invention, is excellent in antimicrobial activity against fungi and general bacteria that cause vaginitis and is suitable for use in the prevention and treatment of vaginitis. In addition, it is excellent in formulations that can be manufactured in various formulations, and it is possible to commercialize products according to the taste and convenience of patients.
Fig. 1 is a photograph showing antibacterial activity against Fusobacterium nucleatum bacterium. [1) Blank 2 占 퐂 / 20 占 퐇 of chloramphenicol 3.42 占 퐂 / 20 占 봉 bee venom 20 占 퐂 / 20 占 봉 bee venom 10 20 실시 Example 1 20 실시 Example 4 20 占 ⑧ Example 3 20 占 ⑨ Comparative Example 2 20 占 퐇.
Fig. 2 is a photograph showing the antibacterial activity against Proteus mirabilis [Blank 2 占 퐂 / 20 占 퐇 of chloramphenicol 3.42 占 퐂 / 20 占 퐇 of beampox 20 占 퐂 / 20 占 봉 bee venom 10 占 퐂 / 20 ⑥ Example 1 20 占 실시 Example 4 20 占 ⑧ Example 3 20 占 ⑨ Comparative Example 2 20 占 퐇.
FIG. 3 is a transmission electron micrograph (12,000 times, N: nucleus, H: hydrogenosomes, V: fear) observed after a control group without control of bee venom (control) was cultured for 24 hours.
FIG. 4 is a transmission electron microscope photograph (12,000 times, A: autogranular body) observed after treating bee venom at 37.5 ppm for 30 minutes.
The present invention relates to a composition for preventing and treating vaginitis comprising bee venom as an active ingredient, and a vaginal detergent.
The vaginitis is Candida albicans, bacterial vaginitis, Trichomonas vaginitis or atrophic vaginitis. Candida albicans may be the causative organism of Candida albicans, and it is also a bacterial vaginitis. The causative organisms of bacterial vaginitis include Gardnerella vaginalis, Mycoplasma spp, Mobiluncus spp, Bacteroides spp, Streptococcus spp, Staphylococcus spp, Anaerobic bacteria such as Staphylococcus aureus, and anaerobic bacteria such as tuberous anaerobic bacteria. Trichomonas vaginitis is caused by a parasitic infestation of trichomoniasis, and atrophic vaginitis can be caused by a decrease in vascular mucosal thickness after menopause. In this case, Megasphaera Atopobium vaginae, Eggerthella spp, Clostridium-like spp, Peptostreptococuus micros, Prevotella spp., Pseudomonas spp. spp), and the like, and anaerobic bacteria.
The beeing used in the present invention will be described in detail as follows.
Bee venom (BEE-VENOMS) is a toxic liquid from the honey bee's ovipositor, which is composed of various components, among which peptides are known to play an anti-inflammatory and antimicrobial action, strong analgesic action and immunity enhancement.
The ingredients and efficacy of bee venom will be described in detail as follows.
Treatment of immune system diseases: Bee venom is effective in the treatment of most immune diseases, because bee venom stimulates the immune system of mammals to successfully fight disease. In other words, the two main immune functions of bee venom are to stimulate the organism's biological system and to increase the defenses of the organism.
② Neurotoxic effect: Bee stings cause pain and inflammation, and the substances that cause them are effectively used in mammalian animals in the development of a sparse response, ie, an analgesic. Especially, it shows analgesic action by action of apamin in bee venom. It shows strong analgesic effect on various neuralgia, arthritis, rheumatoid arthritis, gout and myalgia, and its therapeutic effect is also high.
③ Hemolysis: One of the most outstanding effects of the needle action is hemolysis. Melittin, phospholipase, etc., absorb and excrete blood vessels such as bruise, internal bleeding, etc., and supply new blood, oxygen and nutrients to tissues to heal and hemolyze.
④ vasodilating action: the effect of histamine substances, capillaries, small arteries, pancreas, especially the effect of expanding the internal blood vessels is significant. Therefore, diseases such as poor circulation, frostbite, and muscle pain are treated.
⑤ Blood pressure lowering action: Histamine substance acts to lower blood pressure by action. In particular, even at a concentration of 250 millionths of a minute, there is a hypotensive effect, and there is an excellent effect in the treatment of essential hypertension and the like.
⑥ Autonomic control: When the human body receives stress, the autonomic nerves of the sympathetic nerve and the parasympathetic nerve are hunted, which causes many diseases. Catecholamine and acetylcholine, which are substances necessary for normalization of the autonomic nerves, are contained in bee venom. This substance is effective for the treatment of psychosomatic menopausal stress disorder due to the brain cell transporter.
The present invention contemplates a pharmaceutical composition and vaginal detergent for vaginitis prevention and treatment using bee venom which is a natural substance having the above-mentioned effect.
Specifically, in the present invention, the antifungal effect on the fungal pathogenic fungus causing vaginitis and the bee venom showing the antimicrobial effect on the bacteria are contained in an amount of 0.0001 to 10% by weight, preferably 0.01 to 5% By weight. If the bee venom is contained in an amount less than 0.0001% by weight based on the total weight of the composition, the antifungal or antimicrobial effect is low and the efficacy can be reduced. If the bee venom is contained in an amount exceeding 10% by weight, cytotoxicity may be caused.
When such a composition containing bee venom is used as a pharmaceutical composition for prevention and treatment of vaginitis, it is possible to use pharmaceutically acceptable carriers, excipients, dilutes, stabilizers, surfactants, thickeners, They can be mixed and formulated into external preparations such as liquids, gels, suppositories, aerosols and the like.
When such a composition containing bee venom is used as a cosmetic, it can be used as a vaginal detergent by mixing with a carrier, a forming agent, a dilute, a stabilizer, a surfactant, an increasing agent, a pH adjusting agent and the like.
When the composition is formulated with a liquid detergent in the present invention, the preparation is diluted with a solvent such as water at a predetermined ratio, and a surfactant is used in the excipient. The surfactant that can be used at this time is an anionic surfactant, Sulfuric acid, sulfates, sulfone, cationic surfactants, quaternary ammonium compounds, amphoteric surfactants, amino acid-type amphoteric surfactants and betaine-type amphoteric surfactants. Nonionic surfactants include polyhydric alcohol higher fatty acids, polyethylene glycol higher fatty acids, polyoxyethylene higher alcohols, sorbitan higher fatty acids, and polyoxyethylene sorbitan higher fatty acids. In the industry, sulfonic acid, an anionic surfactant, is mainly used as a surfactant, and laurylsulfuric acid and salts thereof are mainly used. Surfactants obtained from natural oils may also be used. Natural oils such as coconut oil, palm oil, cottonseed oil and olive oil are examples of natural oils. Examples of animal oils include tallow, lard, acid oil, light oil and fish oil. In the present invention, a surfactant derived from a natural oil, which is not a surfactant obtained from petroleum or coal, is used to enhance human fitness. As the diluent, poly (ethylene glycol), polypropylene, glycerin, butylene glycol and the like can be used. As the solvent, water, ethanol and the like can be used. In order to improve the sensory properties, fragrance and the like may be additionally used. In order to enhance the stability of the preparation, a stabilizer may be used. As the stabilizer, sodium edetate, ammonium glycyrrhizin and the like may be used.
Other components besides bee venom can be used within a range that does not deteriorate the effect of the present invention as needed. Examples of the present invention include beech poisoning in an amount of 0.0001 to 10% by weight; 10 to 30% by weight of a surfactant; 0.001 to 3% by weight of stabilizer; A cleaning agent containing 60 to 95% by weight of a solvent can be prepared.
In addition, the detergent may be in the form of a spray, a liquid, a gel, a powder or a solid soap, a foam or a wet tissue.
The gel agent in the present invention can be defined as a gel-like (semi-circular) external preparation made of organic molecules having a large molecular weight permeating a liquid to be applied to the skin. The gels include aqueous gels and oily gels. Typically, cellulose derivatives such as hydroxypropylcellulose and the like are usable as polymeric materials used for preparing gels; Natural gums such as xanthan gum, alginate, gum arabic and the like; Other polymeric materials may be carbomers, povidone, and the like. The diluent may be polyethylene glycol, propylene glycol, glycerin, butylene glycol, and the like. Examples of the stabilizer include sodium edetate, ammonium glycyrrhizin, potassium dipotassium glycyrrhizate, and the like. . In addition, gradually increasing triethanolamine and the like can be used, and a pH adjusting agent (citric acid, sodium citrate, etc.) and fragrance can be additionally used.
Other components besides bee venom can be used within a range that does not deteriorate the effect of the present invention as needed. Examples of the present invention include beech poisoning in an amount of 0.0001 to 10% by weight; 0.5 to 15% by weight of a polymeric material; 0.001 to 3% by weight of stabilizer; 80 to 97% by weight of a solvent; 0.5 to 10% by weight of a diluent; A gel containing 0.1 to 5% by weight of the thickening agent can be prepared.
In the present invention, polyethylene glycol having an average molecular weight of 1,000 to 10,000 is used as a base when melamine is formulated as a suppository and melted by the body temperature of the human body to release the bee venom as an active ingredient. In addition, a pH adjusting agent (citric acid, sodium citrate, , Surfactants, stabilizers, and the like.
Other components besides bee venom can be used within a range that does not deteriorate the effect of the present invention as needed. Examples of the present invention include beech poisoning in an amount of 0.0001 to 10% by weight; Suppositories containing 90 to 99.999% by weight of a base can be prepared.
In the present invention, it is possible to prepare beverages containing surfactants, solvents, pH adjusters, stabilizers and fragrances in the form of an aerosol agent which releases the fine dispersion of the liquid or solid material containing the active ingredient under the action of gas pressure have. The diluent may be polyethylene glycol, propylene glycol, glycerin, butylene glycol, etc. The surfactant may be an anionic surfactant, a cationic surfactant, a cationic surfactant, an anionic surfactant, Nonionic surfactants can be used. As the stabilizer, sodium edetate, ammonium glycyrrhizinate, potassium dipotassium glycyrrhizinate and the like can be used, and citric acid, sodium citrate and the like can be used as a pH regulator. Propellant gas, butane gas, compressed air, nitrogen gas and the like can be used as a jetting substance for gas pressure. In the examples of the present invention, a mixture of water, ethanol and butylene glycol was used as a solvent, and citric acid, sodium citrate and the like were used as pH adjusters. The stabilizer was sodium edetate, potassium glycyrrhizinate and ammonium glycyrrhizinate Nitrogen gas was used as a jetting body for gas pressure.
Other components besides bee venom can be used within a range that does not deteriorate the effect of the present invention as needed. Examples of the present invention include beech poisoning in an amount of 0.0001 to 10% by weight; 0.001 to 3% by weight of stabilizer; 85 to 99.7% by weight of a solvent; an aerosol agent containing 0.001 to 1.0% by weight of a pH adjusting agent, 0.1 to 10% by weight of a diluent, and 0.01 to 1.0% by weight of a surfactant can be prepared.
In the present invention, a pharmaceutical composition or a vaginal detergent containing bee venom is excellent in antifungal activity and antimicrobial activity against Candida albicans and general bacterium which are the fungi causing vaginitis and thus inhibits or suppresses the growth and life functions of bacteria and fungi can do.
The pharmaceutical preparations may be formulated as tablets, pills, powders, granules, capsules and the like as solid preparations for oral administration. Examples of the form for parenteral administration include creams, ointments, lotions, dressings, Other coating agents, and the like.
Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. However, it is to be understood that the present invention is not limited to the disclosed embodiments, but may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.
Manufacturing example One: Beijing detach
The honey bee ( Apis meliffera L.) was collected by using a bee venom collecting device, and the collected bee venom was dissolved with distilled water, centrifuged, filtered, and lyophilized to separate the pure bee venom.
Example 1: quality detergent
Bee venom 20.0 mg
15.0 g of coco glucoside (a surfactant derived from coconut oil)
5.0 g of sodium cocoammonoacetate (surfactant derived from coconut oil)
Sodium edetate 100.0 mg
Purified water 79.88 g
The raw materials were weighed, placed in a preparation vessel, and completely dissolved by stirring while heating the solution temperature to 80 占 폚. When completely dissolved, the solution temperature was cooled to 40 DEG C and then degassed under vacuum to prepare 100 g of a quality detergent.
Example 2: Vaginal detergent
Bee venom 100.0 mg
15.0 g of coco glucoside (a surfactant derived from coconut oil)
5.0 g of sodium cocoammonoacetate (surfactant derived from coconut oil)
Sodium edetate 100.0 mg
Purified water 79.80 g
The raw materials were weighed, placed in a preparation vessel, and completely dissolved by stirring while heating the solution temperature to 80 占 폚. When completely dissolved, the solution temperature was cooled to 40 DEG C and then degassed under vacuum to prepare 100 g of a quality detergent.
Comparative Example 1: Vaginal detergent ( Placebo )
15.0 g of coco glucoside
5.0 g of sodium cocoammonoacetate
Sodium edetate 100.0 mg
Purified water 79.90 g
The raw materials were weighed, placed in a preparation vessel, and completely dissolved by stirring while heating the solution temperature to 80 占 폚. Upon complete dissolution, the solution temperature was cooled to 40 ° C and vacuum degassed to produce 100 g of a vaginal detergent placebo.
Example 3: Gelse ( Beijing Concentration: 12.5 占 퐂 / g)
Bee venom 1.25 mg
Sodium edetate 50.0 mg
Potassium glycyrrhizinate 50.0 mg
3.0 g of butylene glycol
Povidone K30 1.0 g
Carbomer 300.0 mg
Triethanolamine 400.0 mg
Purified water 95.19875 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and degassed under vacuum to prepare 100 g of a gel.
Example 4: Gelse ( Beijing Concentration: 25.0 占 퐂 / g)
Bee venom 2.5 mg
Sodium edetate 50.0 mg
Potassium glycyrrhizinate 50.0 mg
3.0 g of butylene glycol
Povidone K30 1.0 g
Carbomer 300.0 mg
Triethanolamine 400.0 mg
Purified water 95.1975 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved while being heated at 80 DEG C with stirring, cooled at 40 DEG C and vacuum degassed to prepare 100 g of a gel.
Example 5: Gelse
Bee venom 100.0 mg
Sodium edetate 50.0 mg
Potassium glycyrrhizinate 50.0 mg
3.0 g of butylene glycol
Povidone K30 1.0 g
Carbomer 300.0 mg
Triethanolamine 400.0 mg
Purified water 95.10 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and degassed under vacuum to prepare 100 g of a gel.
Comparative Example 2: Gelse ( Placebo )
Sodium edetate 50.0 mg
Potassium glycyrrhizinate 50.0 mg
3.0 g of butylene glycol
Povidone K30 1.0 g
Carbomer 300.0 mg
Triethanolamine 400.0 mg
Purified water 95.20 g
The raw materials were weighed, then placed in a preparation vessel, completely dissolved while stirring at 80 DEG C, cooled to 40 DEG C, and degassed under vacuum to prepare 100 g of a gel.
Example 6: Suppository
Bee venom 5.0 mg
Polyethylene glycol 1500 49.995 g
Polyethylene glycol 4000 50.0 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and defoamed in vacuo to prepare 100 g of suppositories.
Example 7: Suppository
Bee venom 100.0 mg
Polyethylene glycol 1500 49.9 g
Polyethylene glycol 4000 50.0 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and defoamed in vacuo to prepare 100 g of suppositories.
Comparative Example 3: Suppository ( Placebo )
Polyethylene glycol 1500 50.0 g
Polyethylene glycol 4000 50.0 g
The raw materials were weighed, placed in a preparation vessel, completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and defoamed in vacuo to prepare 100 g of suppositories.
Example 8: Aerosol agent
Bee venom 5.0 mg
3.5 g of butylene glycol
Ethanol 3.0 g
0.5 g of glycerin
≪ tb > < tb > Pigment-60 Hydrogenate castor oil 0.3 g
Citric acid 20.0 mg
Sodium citrate 80.0 mg
Sodium edetate 50.0 mg
Ammonium glycyrrhizine 10.0 mg
50.0 mg of potassium glycyrrhizin potassium
Purified water 92.485 g
The raw material was weighed and placed in a preparation container. The solution was completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and vacuum degassed to prepare 100 g of a solution. The solution was filled in a pressure vessel, To prepare an aerosol formulation.
Example 9: Aerosol agent
Bee venom 100.0 mg
3.5 g of butylene glycol
Ethanol 3.0 g
0.5 g of glycerin
≪ tb > < tb > Pigment-60 Hydrogenate castor oil 0.3 g
Citric acid 20.0 mg
Sodium citrate 80.0 mg
Sodium edetate 50.0 mg
Ammonium glycyrrhizine 10.0 mg
50.0 mg of potassium glycyrrhizin potassium
Purified water 92.39 g
The raw material was weighed and placed in a preparation container. The solution was completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and vacuum degassed to prepare 100 g of a solution. The solution was filled in a pressure vessel, To prepare an aerosol formulation.
Comparative Example 4: Aerosol agent ( Placebo )
3.5 g of butylene glycol
Ethanol 3.0 g
0.5 g of glycerin
≪ tb > < tb > Pigment-60 Hydrogenate castor oil 0.3 g
Citric acid 20.0 mg
Sodium citrate 80.0 mg
Sodium edetate 50.0 mg
Ammonium glycyrrhizine 10.0 mg
50.0 mg of potassium glycyrrhizin potassium
Purified water 92.485 g
The raw material was weighed and placed in a preparation container. The solution was completely dissolved with stirring at 80 DEG C, cooled to 40 DEG C, and vacuum degassed to prepare 100 g of a solution. The solution was filled in a pressure vessel, To produce an aerosol placebo.
Experimental Example 1: Antibacterial effect test ( Agar diffusion assay )
The paper disc method was performed during the agar diffusion assay in order to examine the antimicrobial effect of the composition for the prevention and treatment of vaginitis-containing vaginitis and the vaginal detergent.
The strains used were Escherichia coli (gram negative), staphylococcus aureus (gram positive), Enterobacter aerogenes (gram negative) and Klebsiella pneumoniae (gram negative) as thermogenic anaerobic strains. Proteus mirabilis (Gram negative) and Fusobacterium nucleatum (gram negative) were used.
The strain used was cultivated in the medium to increase its activity and then used in the experiment. The agar medium was placed in a Petri dish and hardened. The test strain was spread out by spreading uniformly. The circular filter paper was placed on the medium, and a bee venom and vaginal detergent, a gel, and an aerosol agent were added to the medium. The culture was incubated in an incubator at 37 ° C for 18 hours under anaerobic conditions to confirm the generation of inhibition of growth.
As a result, Escherichia coli , staphylococcus aureus , Enterobacter aerogenes , Klebsiella pneumoniae In the case of bee venom and vaginal detergent, it shows antibacterial effect because of the appearance of low circulation. In the case of anaerobic bacteria such as Fusobacterium necleptum and Proteus mirabilis, Respectively.
Anaerobic strain In the case of Fusobacterium necleptum and Proteus mirabilis , the experimental procedure for the round filter paper method was as follows: Bacto® RCM (Reinforced Clostridial Medium) was prepared according to the manufacturer's recommended method, 20 ml was added to a 90 mm Petri dish And then used.
In order to increase the activity of the experimental strains Fusobacterium nucretum and Proteus mirabilis, subculture was carried out in the above culture medium at 35 ° C. for 18 hours under anaerobic conditions. Subcultured bacteria were spread evenly on the above medium and cultured at 35 ° C for 18 hours under an anaerobic condition. Then, 10 ml of 0.9% physiological saline was added to the petri dish to scrape the cultured bacteria to obtain a homogenous broth. Separately, 100 ml of Bacto® RCM medium was prepared and stored at 40-45 ° C to prevent it from becoming hard. To this, 1 ml of the above-mentioned yeast solution was added thereto, homogenized, and 5 ml was quickly dispensed into a petri dish containing the medium. A 6 mm round filter paper was placed thereon. The test solution was incubated at 35 ° C for 18 hours under anaerobic conditions with bee venom, bee venom, and Examples 1 to 3 and Comparative Example 2, and then examined for inhibition of the antibacterial activity. Respectively. Figure 1 shows the Fusobacterium nocleptum , Figure 2 shows The experimental results of Proteus mirabilis are shown.
Experimental Example 2: Antifungal effect test ( Agar diffusion assay )
The antifungal effect of bee venom, vaginal cleanser, gel, suppository, and aerosol was investigated by using Candida albicans, an indicator strain of Candida albicans albicans ) were used.
The strains used were cultured in medium to increase activity and then used in the experiment. The agar medium was placed in a Petri dish and hardened. Then, the test strain was spread out by spreading method. The circular filter paper was placed on the medium, and a bee venom, a vaginal detergent, a gel agent and an aerosol agent were added to the medium, And then cultured under aerobic conditions at 25 ℃ for 5 days.
As a result of the experiment, it was confirmed that the antifungal effect was confirmed by confirming the inhibition against Candida albicans fungus.
Experimental Example 3: Antibacterial effect test ( growth inhibition test )
To verify the antimicrobial effect of bee venom, vaginal detergent, gel, suppository and aerosol agent, the liquid medium was diluted in a test tube and inoculated with a test strain (10 5 to 10 6 CFU / ml) After incubation under the anaerobic conditions, the turbidity of each test tube was observed to confirm the antimicrobial effect.
If there is no antimicrobial action, turbidity will be high. If there is antibacterial action, there will be no turbidity. The test strains used were Escherichia coli (gram negative) , Enterobacter aerogenes (gram negative) , Klebsiella pneumoniae (gram negative) Proteus was isolated from the vagina of anaerobic bacteria mirabilis (gram negative), Fusobacterium nucleatum (Gram negative) was used.
In addition, antibacterial effects of antibiotics such as ampicilline, chloramphenicol, and kanamycin were compared.
In the case of bee venom in the experiment, the bee venom was dissolved in purified water (in terms of bee venom concentration shown in Table 1). In the case of Example 1 and Comparative Example 1, a specimen was diluted 10-fold with purified water. At the time of the test, 20 ㎕ of each strain was placed in a test tube inoculated with the test strain. After culturing in an incubator, the turbidity was measured at a wavelength of 600 nm using a UV / Vis spectrophotometer.
The experimental results are shown in Table 1 below.
50 [mu] g / ml
10 [mu] g / ml
100 [mu] g / ml
170 [mu] g / ml
50 [mu] g / ml
+++: absorbance less than 0.05 antibacterial power very good
++: Absorbance 0.05 ~ 0.20 Good antimicrobial activity
+: Absorbance 0.20 ~ 0.30 Antimicrobial power Normal
-: Absorbance 0.30 or more No antibacterial activity
As a result of the tests in Table 1, bee venom showed the same antibacterial activity as antibiotics such as ampicillin, chloramphenicol, and kanamycin. The vaginal cleaner of Example 1, the gel of Examples 3 and 4, the suppositor of Example 6, The antibiotics showed the same antimicrobial activity as the antibiotics.
Experimental Example 4: Antifungal effect test ( growth inhibition test )
To verify the antifungal effects of bee venom, vaginal cleanser, gel, suppository and aerosol, the liquid medium was diluted and then inoculated with a test strain (10 5 to 10 6 CFU / ml) in an incubator at 30 to 37 ° C and 18 After incubation under the anaerobic conditions, the turbidity of each test tube was observed to confirm the antimicrobial effect. If there is no antimicrobial action, turbidity will be high. If there is antibacterial action, there will be no turbidity. Candida albicans, an indicator strain of Candida albicans, was used as a test strain.
In addition, the effects of fluconazole and clotrimazole antifungal agents were compared.
In the case of bee venom in the experiment, the bee venom concentration in Table 2 was dissolved by dissolving in purified water. In the case of Example 1 and Comparative Example 1, a sample was diluted 10-fold with purified water. At the time of the test, each 20 μl was put into a test tube inoculated with the test strain. After culturing in an incubator, the turbidity was measured at a wavelength of 600 nm using a UV / Vis spectrophotometer.
The experimental results are shown in Table 2 below.
30 mu g / ml
30 mu g / ml
+++: absorbance less than 0.05 Antifungal activity very good
++: Absorbance 0.05 ~ 0.20 Good antifungal effect
+: Absorbance 0.20 ~ 0.30 Antifungal force Normal
-: Absorbance 0.30 or more No antifungal activity
As a result of the tests in Table 2, the bee venom showed antibacterial activity equivalent to that of fluconazole and clotrimazole, which are antibacterial agents, and the vaginal detergent of Example 1, the gel of Examples 3 and 4, the suppositor of Example 6, The antimicrobial activity was similar to that of the antifungal agent, and it was confirmed that the antimicrobial activity was effective in preventing and improving vaginitis.
Experimental Example 5: Survival rate experiment for vaginal mucilage
Survival rate experiments were conducted to confirm the effects of bee venom, vaginal cleanser, gel, suppository, and aerosol agent on the vagal mite.
Trichomonas vaginalis was purchased from ATCC 30001, and TYM medium (Trypticase-yeastextract-maltose medium) of Diamond (Table 3) was used. The vagotrophs were placed in a 15 ml glass culture tube containing 5 ml of TYM medium and cultured in a 5% CO 2 incubator at 37 ° C for 24 hours. The cells were stained with 0.4% trypan blue to measure the number of viable cells Respectively.
3 × 10 5 cells were cultured in 5 ml of TYM medium at 37 ° C. in a 5% CO 2 incubator for 23 hours and 30 minutes. Subsequently, the vaginal detergent, gel, Ml, and then incubated for another 30 minutes. The vaginal mites were separately collected from each glass tube and the separated vaginal mites collected were washed once with PBS. After adding the new culture medium, the mixture was mixed and trypan blue was well mixed. Then, the mixture was put into a hemocytometer and the number of living cells was measured using an optical microscope. Separately, bee venom was diluted in purified water at a concentration of 100 μg / ml, purified water, and metronidazole, which is a drug used for treatment of trichomonias, were diluted in purified water to a concentration of 100 μg / ml. . The experimental results are shown in Table 4 below.
(100 쨉 g)
(100 mu g / ml)
+++: Suppresses more than 90%, and has excellent antigenicity
++: 60 ~ 90% suppression, excellent antigenic effect
+: 10 to 60% inhibition, antigen-positive
- : no effect
As shown in Table 4, bee venom showed the same antigenic potency as that of metronidazole, which is an antitricomonas therapeutic drug. The vaginal detergent of Example 2, the gel of Example 5, the suppository of Example 7, It was also confirmed that antibiotics were effective in preventing and improving Trichomonas vaginitis.
Experimental Example 6: Transmission electron microscope ( TEM )
For the sections for transmission electron microscopy, the test group treated with bee venom was collected after culturing and centrifuged. After centrifugation, the supernatant was removed and fixed in 2% paraformaldehyde-2.5% glutaraldehyde (pH 7.2) solution buffered with 0.1 M phosphate buffer for 4 hours, then washed with PBS and further washed with 2% osmium tetraoxide for 1 hour And then dehydrated with ethanol. After replacing with propylene oxide, Epon 812 was solidified. After polymerization for 3 days at 60 ° C in a polymerizer, the solidified tissue was stained with alkali toluidine blue and observed with a transmission electron microscope.
Transmission electron microscopy showed that the bee venom was treated at 37.5 ppm for 30 minutes and the microstructural changes in Trichomonas vaginalis cells were observed. The nuclear membrane was contracted and irregular shape was observed in the cell membrane. The number of follicles was increased, and the induction of vaginal phloem mortality was confirmed by the formation of autogranular bodies [Fig. 4].
Claims (8)
Wherein the vaginitis is bacterial vaginosis, Candida vaginitis, Trichomonas vaginitis, or atrophic vaginitis.
Wherein the bee venom is 0.0001 to 10% by weight based on the total weight of the composition.
Spray type, solid type, liquid type or gel type soap; Foam; Or a vaginal detergent in oil form.
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| KR20130031400 | 2013-03-25 | ||
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| KR1020140006132A KR20140116788A (en) | 2013-03-25 | 2014-01-17 | Composition for protecting and treating vaginosis disease comprising bee venom effective for treatment of vaginitis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3796981A4 (en) * | 2018-06-28 | 2022-03-30 | Chemistryrx | Compositions and methods for treating trichomonas |
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2014
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3796981A4 (en) * | 2018-06-28 | 2022-03-30 | Chemistryrx | Compositions and methods for treating trichomonas |
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