KR101198572B1 - Composition for treatment or improvement of microbial infection - Google Patents

Abstract

The present invention is anaerobic containing steam distillation extract extracted from one or more mugwort selected from Artemisia princeps, Artemisa capillaris and Artemisia iwayomogi as an active ingredient. Microorganisms comprising one or more microorganisms for treating or improving microbial infections selected from bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans, pharmaceutical compositions for treating vaginitis, detergent compositions for improving vaginitis, and microorganisms containing 1-octen-3-ol as active ingredients At least one microorganism selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans, including at least one of a composition for treating or improving an infection, a pharmaceutical composition for treating yeast infection, a cleaning composition for improving vaginitis, and at least one selected from alpha-terpineol and eugenol. Composition for the treatment or improvement of infectious disease , Provides a pharmaceutical composition, a cleaning agent for improving salt composition for treating vaginitis. The composition of the present invention can inhibit anaerobic bacteria Atopobium vaginae, Gardnella vaginalis and / or fungi Candida albicans, and is effective in treating or improving vaginitis. In addition, the growth of lactic acid bacteria, which is a beneficial bacterium, is not significantly affected.

Description

Composition for treatment or improvement of microbial infection

The present invention relates to a composition for treating or ameliorating at least one microbial infection selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans, and more particularly, to a composition effective for treating and / or improving vaginitis.

Mugwort is an asteraceae plant widely distributed in northern hemisphere including Asia, Europe, USA, about 400 species are known, and about 30 species grow in Korea. The varieties include Fine Artemisia subulata Nakai / A. stenophylla Kitamura, Artemisia annua Linne, Artemisia aureum Artemisia apiaceae, Cloud mochi; Anaphalis sinica Hance ssp. morii (Nakai) Kitamura, Japanese wormwood; Artemisa laciniata Willd, Artemisia sylvatica Maximowicz, Artemisia sylvatica Maximowicz, Artemisia aurata, Euphorbia Artemisia aurata, Artemisia stolonifera (Max.) Komarov, Artemisia schmidtiana , Moth Gnaphalium affine D. Don, Mugwort Artemisia keiskeana, Mugwort Artemisia selengensis TURCZ, Silkworm Artemisia lagocephala var. triloba, Artemisia scoparia Waldstein et Kitamura, Artemisia Artemisi feddei Leveille et Vaniot, Artemisia feddei Leveille et Vaniot, Tarragon Artemisia capillaris Thunberg viridis Bess, Artemisia hallaisanensis Nakai, Real Artemisia japonica var. angustissima, Artemisia princeps var. orentalis (Pamp.) Hara, wormwood Artemisiae asiatica NAKAI, A. viridissima Pamp. lavandulaefolia DC, Artemisia mongolica Fischer), Sage Artemisia fukudo Makino, Artemisia sacrorum Ledeb. ssp. There are various wormwoods, such as manshurica Kitamura, Artemisia argyi Leveille et Vaniot, and Yellow Wormwood Artemisia stelleriana Bess. In addition to this, it is known that the indigenous wormwood, Seombi wormwood, white heather wormwood, white mountain wormwood, and foliar mulberry. Among them, Artemisia princeps Pamp. Is a wormwood distributed in Ganghwa-do.

The thirty species of mugwort in Korea show significant differences between the external form and the ingredients they contain, and their use (efficacy) is also different.

Mugwort is the first of its kind in the city written by Chinese national era (BC 403-221). It is assumed that it has been used since this time. Based on the use of cheongho in Chinese medicine, it is used to separate the antimalarial artemisin from A. annua and to develop styrene, which is an extract of Artemisia asiatica (DA-9601). Doing.

In addition, in the private sector, hemostatic drugs are used for postpartum bleeding, nosebleeds, uterine bleeding, etc. In addition, tonic blood, gynecological health, and diarrhea treatment are used.

Mugwort has been developed into a variety of health food or cosmetics. There are perfumes, preservatives, antibacterial cosmetics, sanitary napkins, moxa, fumigation baths and bathing agents in cosmetics. The mugwort used for this purpose is A. asiatica, mostly used in herbal medicine.

On the other hand, vaginitis is a gynecological disease that most women have with cold and hypothalamus, itching, tingling, pain during sexual intercourse, tingling and burning or odor during urination. The most common infectious vaginitis is bacterial vaginitis and Candida vaginitis. Bacterial vaginitis is caused by the anaerobic strains Atopodium vaginae and / or Gardenella vaginalis strains. In addition, the aerobic strains Staphylococcus aureus and Escherichia coli are infected around the entrance of the vagina. Therefore, in order to fundamentally treat vaginitis, the anaerobic strains Atopobium vaginae and Gardenella vaginalis must be eliminated.

Fungal vaginitis is most often caused by Candida albicans. However, fungal vaginitis can be improved with azole antifungal agents unlike bacterial vaginitis, but the development of new drugs is required because the disease cannot be eliminated by the antimicrobial agent.

The antimicrobial agents used in these infections differ in their efficacy according to the type of bacteria, and antimicrobial agents that simultaneously suppress the causative bacteria of bacterial vaginosis and candidiasis vaginitis are not common. For example, penicillin kills staphylococcus, but not the fungus Candida. However, azole preparations such as itraconazole kill fungi such as candida but do not kill bacteria such as staphylococci.

The present inventors have completed the present invention by studying natural drugs effectively inhibiting Atopobium vaginae and Gardenella vaginalis bacteria and Candida albicans fungi which cause various vaginitis.

One problem to be solved by the present invention is to provide an effective composition for treating or ameliorating one or more microbial infections selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans, which are vaginal agents.

Another object of the present invention is to provide a pharmaceutical composition for treating vaginitis.

Another object of the present invention is to provide a cleaning composition for vaginitis improvement.

The present invention comprises steam distillation extract extracted from at least one mugwort selected from Artemisia princeps, Artemisa capillaris, and Artemisia iwayomogi as an active ingredient. It provides a composition for treating or ameliorating one or more microbial infections selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans.

The Artemisia princece includes lion cucurbita and / or saquay cucurbita, and the like, preferably lion cucurbita.

The steam distillation extract refers to an extract obtained by passing water vapor through mugwort and condensing the passed water vapor. The steam distillation extract is preferably a pressurized steam distillation extract, the pressurized water vapor means a water vapor having a saturated steam pressure greater than the atmospheric pressure. When the water is heated to a temperature of over 100 degrees Celsius under atmospheric pressure in a sealed condition, the saturated steam pressure becomes larger than the atmospheric pressure and becomes pressurized steam. The steam distillation extract may be prepared using a conventional steam distillation extraction apparatus, it is possible to produce a pressurized steam distillation extract using the apparatus described in Korean Patent No. 10-0820344.

The pressurized steam distillation extract is preferably in trans-sabinene hydrate, cis-sabinene hydrate, terpinene-4-ol, beta-caryophyllene, 1-octen-3-ol, alpha-terpineol, germacrene D, valencene, and cryophylene oxide It may include one or more active ingredients selected.

The steam distillation extract is not limited to this, but preferably may be distilled by passing through the mugwort with steam heated to 105 ~ 110 degrees Celsius in a closed container.

The water vapor distillation extract of the active ingredient in the composition of the present invention may include at least one selected from Eucalyptol, borneol, alpha-terpineol, 1-Octen-3-ol and eugenol as an active ingredient, the active ingredient is 50 of the total extract v / v% or more.

The active ingredient in the composition of the present invention is preferably for the treatment or improvement of anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans infection, and can effectively inhibit bacteria and fungi, which are the causative agents of vaginitis, and is effective for various vaginitis.

The therapeutic or ameliorating composition may be a pharmaceutical composition or a detergent composition, and the detergent may be a vaginal cleaner.

The extract of the composition of the present invention is excellent in inhibiting the anaerobic bacteria Atopobium vaginae and / or Gardnella vaginalis and / or fungi Candida albicans, which have an effect on vaginitis when inhibiting growth, the present invention is effective Atopobium vaginae and / or Gardnella vaginalis and / or fungi Candida albicans infectious diseases, or treating or improving vaginitis.

Accordingly, the present invention provides a pharmaceutical composition for treating vaginitis comprising the extract of the composition of the present invention as an active ingredient.

The present invention also provides a cleaning composition for vaginitis improvement comprising the extract of the composition of the present invention as an active ingredient. The cleaner may preferably be a vaginal cleaner.

The present invention also provides a composition for treating or ameliorating at least one microbial infection selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans containing 1-octen-3-ol as an active ingredient.

Preferably, the microorganism may be anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans.

In addition, the present invention provides a pharmaceutical composition for treating vaginitis comprising 1-octen-3-ol as an active ingredient.

The present invention also provides a cleaning composition for vaginitis improvement comprising 1-octen-3-ol as an active ingredient.

In another aspect, the present invention provides a composition for treating or ameliorating at least one microbial infection selected from anaerobic bacteria Atopobium vaginae and Gardnella vaginalis containing at least one selected from alpha-terpineol and eugenol as an active ingredient.

The present invention also provides a pharmaceutical composition for treating vaginitis containing at least one selected from alpha-terpineol and eugenol as an active ingredient.

The present invention also provides a cleaning composition for vaginitis improvement comprising at least one selected from alpha-terpineol and eugenol as an active ingredient.

In the present invention, vaginitis may be fungal vaginitis and / or bacterial vaginitis.

The active ingredient included in the composition of the present invention may be preferably extracted from Artemisia princeps (Artemisia princeps).

The composition for treatment or improvement of the present invention may be a pharmaceutical composition or a detergent composition, and the detergent may be a vaginal cleaner.

The active ingredient and / or active ingredient in the composition of the present invention may be 0.5 to 20% by weight of the total weight.

In addition, the present invention relates to the anaerobic bacteria Atopobium vaginae and Gardnella vaginalis, comprising administering to a mammal (including human) in need of pharmaceutically effective amount of the active ingredient and / or active ingredient in the composition of the present invention. The present invention provides a method for treating or ameliorating at least one infectious disease selected from the fungi Candida albicans, a treatment for vaginitis, and a method for improving vaginitis.

Unless otherwise stated, the content of the composition of the present invention is equally applied to the composition and composition of the present invention as long as it does not contradict the use and method of the active ingredient and active ingredient.

The composition, the active ingredient, and the active ingredient of the present invention can be administered orally or parenterally to a mammal including a human, and are formulated by combining the active ingredient or active ingredient with a pharmaceutically or cosmetically acceptable carrier. May be administered.

When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants which are commonly used may be used. Formulations for parenteral administration include suppositories and the like, and suppositories may include witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, and / or gelatin.

In addition, a pharmaceutically acceptable emulsifier, diluent, excipient, or the like may be added to the composition, the active ingredient, and the active ingredient of the present invention, or a base material for cosmetic preparation, but is not limited thereto, but a solution, a suspension , Emulsions, pastes, gels, creams, lotions, powders, soaps, cleansing, foundations, and sprays. The pharmaceutical dosage form may be an external preparation, and may preferably be an external preparation for skin such as a spray, an ointment, a warning, a lotion, a lining, a pasta, or a cataplasm. The composition of the present invention, the active ingredient, or the active ingredient in the composition can be applied to the external application for spraying, applying or washing the applied site to 0.01 ~ 10mg / time, 1 to 6 times / day application site on an adult basis.

For formulation, reference may be made to Remington's Pharmaceutical Science (Recent), Mack Publishing Company, Easton PA, and others.

The composition of the present invention can inhibit anaerobic bacteria Atopobium vaginae, Gardnella vaginalis and / or fungi Candida albicans and are effective in treating vaginitis (improvement). In addition, the growth of lactic acid bacteria, which is a beneficial bacterium, is not significantly affected.

1 is a result of GC-MS analysis of a preparation example.

Hereinafter, the present invention will be described in more detail with reference to Examples and Preparation Examples, but the following Examples and Preparation Examples are only for illustrating the present invention, and the content of the present invention is not limited to the following Examples or Preparation Examples.

Preparation Example 1 Preparation of a Steam Distillation Extract Boiled to 105 Degrees Celsius Under Pressurization

Artemisia Princeceps (Saint Wormwood Wormwood); Korea, Ganghwado} 500g into a steam distillation apparatus (EssenLab-plus, Hanil Lab Tech, Korea), and through the mugwort with steam heated to 105 degrees Celsius in an airtight container, the steam is room temperature (10-25 degrees Celsius) The mixture was cooled to) to obtain an essential oil layer to prepare a water vapor distillation extract.

Preparation Example 2 Preparation of a Steam Distillation Extract Boiled to 110 Degrees Celsius Under Pressurization

Pressurized steam distillation extract was prepared in the same manner as in Preparation Example 1, except that steam was boiled in a closed container to 110 degrees Celsius instead of 105 degrees Celsius.

Preparation Example 3 Preparation of Water Vapor Distillation Extract

A steam distillation extract was prepared in the same manner as in Preparation Example 1, except that steam was heated to 100 degrees Celsius instead of 105 degrees Celsius.

Reference Example 1 Preparation of Ether Extract

After extracting 1 kg of raw lionwood mugwort in 5 liters of ether at room temperature overnight, the mixture was filtered with filter paper and concentrated with a rotary pressure reducer (Sunil Isla, Eyla KSB201, Korea) to prepare an ether extract.

Reference Example 2 Alcohol Extract Preparation

1 kg of green moths were dried and refluxed in a water bath at 100 degrees Celsius for 2 hours in 5 liters of 80% ethanol, and the supernatant was concentrated under reduced pressure to prepare an alcohol extract.

Reference Example 3 Preparation of Water Extract

1 kg of raw lionwood mugwort was extracted under reflux in a water bath at 100 ° C. for 2 hours with 5 liters of water, filtered through a filter paper, and the supernatant was concentrated under reduced pressure to prepare a water extract.

Reference Example 4 Preparation of Lavender Steam Distillation Extract

A lavender distillate extract was prepared in the same manner as in Preparation Example 1, except that lavender (Herb Mall, Korea) was used instead of Artemisia princecept.

<Comparative Example 1> Preparation of methanol extract of mugwort

0.5 kg of Korean mugwort (Artemisia mongolica Fischer, Gyeongdong Market, Korea) was dried and refluxed in 2.5 liters of methanol for 2 hours in an 80 ° C water bath, and the supernatant was concentrated under reduced pressure to prepare a methanol extract.

<Comparative Example 2> Preparation of Ethanol Extract of Mugwort

0.5 kg of Korean mugwort (Artemisia mongolica Fischer, Gyeongdong Market, Korea) was dried and refluxed in 2.5 liters of 80% ethanol for 2 hours in a 100 degree Celsius water bath, and the supernatant was concentrated under reduced pressure to prepare a methanol extract.

Comparative Example 3 Preparation of Acetone Extract of Mugwort

0.5 kg of sesame seeds (Artemisia mongolica Fischer, Gyeongdong Market, Korea.) Was dried and extracted by 2.5 liters of acetone by cooling for 24 hours (room temperature), and the supernatant was concentrated under reduced pressure to prepare acetone extract.

Experimental Example 1 Extraction Yield Calculation

The yields of the extracts of Preparation Examples 1 to 3 and Reference Examples 1 to 3 were calculated, and the results are shown in the following table. Calculation of the yield was calculated by the amount obtained by extraction compared to the amount used for extraction.

[Table 1]

As a result, it can be seen that the yield of the pressurized steam extract is significantly greater than the other extracts.

From this result it can be seen that the extract of the present invention is a high yield extract.

Experimental Example 2 GC-MS Analysis of Preparation Example 2 Extract

Under the following conditions, GC-MS analysis of the extracts of Lion cucurbita prepared in Preparation Example 2 was performed, and the results are shown in FIG. 1 and the following table. Figure 1 shows the results of GC-MS analysis, the x-axis shows the retention time (minutes), the y-axis shows the height of the peak.

<GC-MS Analysis>

-Analysis equipment: GC / MS Varian GC3800, MS Saturn1200

-Analysis condition: Column VF-5MS

             70 ° C 1 minute, 250 ° C 6 minutes (raised temperature 10 ° C / min)

             Injection vol. 2 μl, temp. 200 ℃, split ratio 50: 1

[Table 2]

It was confirmed that the ingredients described in the above table was included in the extract of Preparation Example 2.

Example 1 Extract Antimicrobial Activity

Transplantation of Provotella bivia (Atopobium vaginae) (KCTC Biological Resources Center, Korea Institute of Bioscience and Biotechnology, Korea) to GAM (general anaerobic medium) medium (Nissui Pharmaceutical Co., Ltd., Japan), Gardnerella vaginalis (KCTC Biological Resource Center) , 5mm Watman sterile, respectively, which were implanted with a transplant of Candida albicans (KCTC Bioresource Center, Korea Institute of Bioscience and Biotechnology, Korea), with a transplant of Saberaud dextrose agar medium (Difco, USA). Place 5 disks evenly and evenly. A 50 ul sample is aspirated (dropped) twice into the filter disc. The cells transplanted into GAM medium were anaerobicly 48 hours at 37 degrees Celsius, and those transplanted into the remaining medium were cultured at 37 degrees Celsius for 24 hours. And the size (diameter) of the blocking zone in which the transplanted strain did not grow was measured. Specimens are extracts of Preparation Examples 1-3 and Reference Examples 1-3, Lavender pressurized steam distillation extract of Reference Example 4 and eucalyptol (another name 'cinneol') which is the main component of lion's foot extract extracted by steam distillation ( Sigma, USA) or Borneol (Sigma, USA), and the control drug Tetracycline (Sigma, USA) or cotrimoxazole (Sigma, USA) was used. In addition, the mugwort vapor pressurized extract prepared in the same manner as in Preparation Example 2 and the extracts of Comparative Examples 1 to 3 were used as specimens except that the mugwort shown in Table 4 was used instead of Artemisia Princecep.

The results are summarized in Tables 3 to 5 below.

[Table 3]

As a result, compared with water, ether, alcohol extracts, and lavender extracts with known antimicrobial activity, the steam distillation extracts of Artemisia princeps and the anaerobic bacteria Atopobium vaginae and Bifidobacterium longum and Lactobacillus acidophilus are not significantly inhibited. Gardnella vaginalis and the fungus Candida albicans are effectively inhibited. In addition, the eucalyptol and Borneol as the main components of the steam distillation extract does not have a large antibacterial activity, it can be seen that the components contained in the pressurized steam distillation extract have a high antibacterial activity. Thus, pressurization containing one or more selected from trans-sabinene hydrate, cis-sabinene hydrate, terpinene-4-ol, beta-caryophyllene, 1-octan-3-ol, alpha-terpineol, germacrene D, valencene, and cryophylene oxide Water vapor distillation extract is thought to be effective in the inhibition of anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans.

[Table 4]

Artemisia princecept extract of various mugwort extracts can be seen that the best inhibit the bacteria and fungi, and the jinjin mugwort, heat wormwood mugwort has an effect on both bacteria and fungi. On the other hand, mugwort had no effect on both vaginal infection bacteria and fungi. Therefore, Artemisia princeps pressurized steam extract, in particular, can be seen to have a significant effect on the infection caused by the anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans. In particular, it can be seen that the inhibitory effect of Candida albicans is 6.76 times (Asiatica Princecep Lowland Area / American Locust Lowland Area = (3.14 * 1.3 ² ) / (3.14 * 0.5 ² ) = 6.76).

[Table 5]

Various solvent extracts of Korean mugwort did not effectively inhibit the anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans.

Therefore, steam distillation extracts of Artemisia Princece, Artemisia capillaris, Artemisia Iwamomogi among various mugwort can effectively inhibit anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and fungi Candida albicans, It can be seen that it is effective.

<Example 2> antimicrobial activity check for each component

Antimicrobial activity confirmation experiment was carried out in the same manner as in Example 1, except that the components shown in Table 6 were used as the specimen instead of the specimen of Example 1. Tetracycline or cotrimoxazole was used as a control drug. The ingredients and the control drugs shown in Table 6 were purchased from Sigma (USA).

The results are summarized in Table 6 below.

TABLE 6

It can be seen that alpha-terpineol, 1-octen-3-ol, eugenol and the like in Table 6 effectively inhibit the anaerobic bacteria Atopobium vaginae and Gardnella vaginalis and the fungi Candida albicans. The above components are particularly effective in treating vaginitis (improvement) because they effectively inhibit Atopobium vaginae, Gardnella vaginalis and Candida albicans without significantly inhibiting the beneficial bacteria Bifidobacterium longum and Lactobacillus acidophilus.

In addition, it can be seen from the above results that eucalyptol and Borneol, which are the main components of Artemisia Princeceps, which are extracted by steam distillation, do not have high antibacterial activity, and are not pressurized rather than the main components extracted by steam distillation. The antibacterial activity of the components extracted by water vapor seems to be large. Therefore, trans-sabinene hydrate, cis-sabinene hydrate, terpinene-4-ol, beta-caryophyllene, alpha-terpineol, germacrene D, valencene, and / or cryophylene oxide contained in pressurized steam distillate extract are considered to be effective.

<Example 3> Antibacterial activity confirmed in vivo

3-1. Antimicrobial Activity in Vaginitis Model Mice Implanted with Candida albicans

Female mice (BALB / c, 17-25 g) were subcutaneously injected with beta-estradiol 17-benzoate (Sigma, MO, USA) dissolved 0.125 mg in olive oil. After 3 days, 5x10 ⁶ Candida albicans (KCTC Bioresource Center, Korea Institute of Bioscience and Biotechnology, Korea) are implanted into the vagina. Divided into groups, for each group, 10% pressurized refinery (Preparation Example 2), 10% distilled refinery (Preparation Example 3), 10% eugenol, 10% 1-Octen once daily for three days thereafter, as shown in the following table. -3-ol, 10% α-Terpineol, 10% Eucalyptol, 10% Borneol, 10% Cotrimazole solution (suspension in 0.01% tween 80 and 1% triton X-100) is administered intravaginally. After 24 hours, beta-estradiol 17-benzoate in 0.125 mg of olive oil was subcutaneously injected for each group. 24 hours and 48 hours later, the vaginal rinse with 0.5 ml of sterile saline solution. The supernatant was diluted 10-fold with 1% peptone water and transplanted into Sabouraoud-dextrose agar containing 0.1% tetracycline (Difco Co., USA), and cultured for 48 hours. In addition, vaginal tissues were isolated and measured for myeloperoxidase activity.

To measure myeloperoxidase (MPO) activity, first place vaginal tissue in 1 ml of 50 mM phosphate buffer (pH 6.0), sonicate, freeze and dissolve three times, and centrifuge to use the supernatant as an enzyme solution. 398 μl of o-dianisidine (0.129 mg / ml) was added to 100 μl of enzyme solution, and H ₂ O ₂ was added at a final concentration of 0.0005%. UV-spectrometer (Jasco system V-530, Japan) at 25 ° C and 492 nm as time course. Activity was measured by One unit of MPO activity is meant to decompose hydrogen peroxide 1mmole / min at 25 ℃. Protein content was measured by Bradford method using bovine serum albumin as standard.

The results are shown in the following table.

[Table 7]

Candida strains were not found in the normal group, and Candida was infected by the Candida-infected group, and many bacteria and high inflammatory markers (MPO) were detected. As a result, strains were reduced and MPO, an inflammation marker, was also decreased. Among them, the inhibitory effect was the highest in the group treated with α-Terpineol.

3-2. Antimicrobial Activity of Gardnella vaginalis Infected Mice

Female mice (BALB / c, 17-25 g) were subcutaneously injected with beta-estradiol 17-benzoate (Sigma, MO, USA) dissolved 0.125 mg in olive oil. Three days later, 1x10 ⁸ Gardnella vaginalis (KCTC Bioresource Center, Korea Institute of Bioscience and Biotechnology, Korea) were implanted into the vagina. Divided into groups, for each group, 10% pressurized refinery (Preparation Example 2), 10% distilled refinery (Preparation Example 3), 10% eugenol, 10% 1-Octen- once daily for 3 days as shown in the following table. 3-ol, 10% α-Terpineol, 10% Eucalyptol, 10% Borneol, 10% tetracycline solution (solution suspended in 0.01% tween 80 and 1% triton X-100) is administered intravaginally. After 24 hours, each group was subcutaneously injected with beta-estradiol 17-benzoate dissolved in 0.125 mg of olive oil. Then wash the vagina with 0.5 ml sterile saline solution for 24 hours and 48 hours. The supernatants were serially diluted 10-fold with 1% peptone water and transplanted into Gardnella vaginalis selective medium plates (0.004% gentamycin, 0.003% nalidixic acid, and 0.0002% amphotericin B-containing Columbia blood agar, MBcell, USA). Count colonies directly with the naked eye. The vaginal tissues were also isolated and measured for myeloperoxidase enzyme activity.

To measure myeloperoxidase (MPO) activity, first put vaginal tissue in 1 ml of 50 mM phosphate buffer (pH 6.0), sonicate and freeze three times, and centrifuge to use the supernatant as an enzyme solution. 398µl of o-dianisidine (0.129mg / ml) was added to 100µl of enzyme solution, and H ₂ O ₂ was added at a final concentration of 0.0005%. Activity was measured. One unit of MPO activity is meant to decompose hydrogen peroxide 1mmole / min at 25 ℃. Protein content was measured by Bradford method using bovine serum albumin as standard.

The results are shown in the following table.

[Table 8]

Gardnella vaginalis strains were not found in the normal group, while Gardnella vaginalis was infected with the vaginitis group, and Gardnella vaginalis was infected with a large number of bacteria and high inflammatory markers (MPO). Significantly decreased strain and MPO, an inflammation marker, also decreased. Among them, the inhibitory effect was the highest in the group treated with α-Terpineol.

Claims (20)

Treatment of one or more microbial infections selected from anaerobic bacteria, Atopobium vaginae and Gardnella vaginalis, containing anaerobic bacteria distilled by distillation through Artemisia princeps with steam heated in an airtight container at 105-110 degrees Celsius. Or a composition for improvement. The composition of claim 1, wherein the Artemisia princes are at least one selected from lion cucurbita or sauerkraut. delete delete delete delete delete delete The composition according to claim 1 for the treatment or amelioration of anaerobic bacteria Atopobium vaginae and Gardnella vaginalis infection. It is caused by at least one anaerobic bacterium selected from Atopobium vaginae and Gardnella vaginalis containing steam distilled extract extracted by distillation through Artemisia princeps with steam heated at 105 ~ 110 degrees Celsius in an airtight container. Pharmaceutical composition for the treatment of bacterial vaginitis. delete delete delete delete delete delete delete delete delete delete

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