KR100646593B1 - Composition comprising the extract of phellinus mushroom and lactic acid for protecting and treating vaginal viral diseases and uterine cancer - Google Patents

Abstract

A composition for protecting and treating vaginal viral diseases and uterine cancer comprising the extract of Phellinus mushroom and lactic acid is provided to increase autoimmunity, inhibit proliferation of bacteria and viruses, and maintain the clean state in the vagina of female. The composition for protecting and treating vaginal viral diseases and uterine cancer comprises the extract of Phellinus sps (Berk). Pilat and lactic acid, wherein the composition is liquid or gel; the weight ratio of Phellinus sps (Berk). Pilat extract and lactic acid is 1:10~1; and the Phellinus sps (Berk). Pilat extract is prepared by extracting Phellinus sps (Berk). Pilat with a polar solvent selected from water, C1-C4 lower alcohol and secondary alcohol.

Description

Composition comprising the extract of phellinus mushroom and Lactic acid for protecting and treating vaginal viral diseases and uterine cancer}

1 This is the result of confirming the growth inhibition of Candida albcans bacteria of genoclean ,

2 is an experimental result confirming the growth inhibition of Staphylococcus aureus bacteria of genoclean ,

3 is The results of experiments confirmed the growth inhibition of E. coli bacteria of genoclean .

The present invention relates to a composition containing an antiviral activity Phellinus sps (Berk) Pilat extract and lactic acid, more specifically, the composition of the mushroom extract and lactic acid in accordance with the present invention is a bacterium in the vagina By inhibiting the proliferation of the virus to maintain the cleanliness of the woman's vagina, it can be used as an effective and safe cleaning composition for preventing and treating intravaginal inflammation and cervical cancer.

Human Papiloma Virus (HPV) is a parasitic virus in a woman's vagina that is difficult to treat and does not go away. It is closely related to the symptoms of warts and various malignancies that infect human and mammalian right upper epithelial cells. The DNA of the virus is detected in more than 90% and 80% of cervical cancers, such as warts that spread into the genitals or anus.

To date, more than 70 species have been found. HPV 16, 18, 31, 33, 35, etc. are mainly detected in cervical cancer and epithelial cancer tissues and are classified as high risk. Detected in non-infectious infections and classified as low-risk. Cytologic findings of infection include koilocytosis and pyskeratocytosis. Cocytosis may be accompanied by premalignant lesions including uterine epithelial cancer or uterine cancer as well as stinging.

If infection is observed in the vulva or cervix of the patient through cytogenetic examination, it is important to check for the presence of a higher lesion by vaginal magnification. This method helps to determine the type of virus by examining the cervix and examining abnormal tissue. Patients infected with or suspected of having human papillomavirus results should undergo regular gynecological examination.

Genital HPV infections in sexually active young women are common, and in most cases these lesions degenerate when the infection is removed or when the lesion develops. Only a few infected women progress to highly epithelial tumors, and only some of them further progress to invasive cancer. The molecular events that lead to HPV infection have not been clearly established. Since there was no suitable in vitro system for propagating human papillomaviruses, they did not evolve into the best information about the virus cycle.

On the other hand, more than 90% of vaginitis in females of childbearing age is bacterial vaginosis, Candida vaginitis, Trichomonas vaginitis, and bacterial vaginosis accounts for 40-50%. These bacteria do not proliferate when the woman's vagina is at normal pH (pH 4.5 to 5.1), but easily penetrates when the body is not normal due to overwork, stress, or taking birth control pills. In addition, complications such as early amniotic membrane frequency and early analgesia can occur, so care must be taken and effective treatment is required.

For this reason, we have been treating yeast infections by putting leftover leaves or wormwood and cooling the boiled water. In addition, Western women `` backed '' the water with roses, lavenders (herb bushes) and bergamos (floral spice trees) .By the 16th century, bidets were used for small words. The main site was washed. Nowadays, the 'vaginal cleaner', which can clean only the vulva, has become easier for women. More than 20% of women of childbearing age usually have vaginitis, and often do not feel symptoms. Itching or burning, odors, changes in color and amount of secretions, as well as when symptoms are not used, once or twice use these cleaners can prevent or treat vaginitis. Most of the products released these days are produced with a weak acidity of pH 4.5, which suppresses abnormal bacterial growth and effectively removes various unpleasant odors occurring during menstrual periods, giving a fresh feeling. Ginobetadine (Mundy Pharma) and Ginodex (Youngjin Pharm.) Are representative products. Such detergents effectively eradicate the causative agents of vaginitis, so if used inside the vagina rather than the vulva or excessively used, it can kill even the beneficial bacteria, which is required to develop safe vaginal cleaners.

Sichuan mushrooms are different from China since ancient times, and they grow on the heartwood of hardwoods such as mulberry, elm, aspen and alder, and are mud mushrooms of Hymenochaetaceae A white fungus belonging to the genus Phellinus Ouel.Em.lmaz . In Korea, the situation mushroom refers to the woody mud mushroom ( Phellinus Linteus ). Situation mushrooms have been used for the treatment of gynecological diseases such as uterine bleeding and menstrual irregularities. Pharmacological and medical researches on the pharmacological action of situational mushrooms have been known to activate immune function following chemotherapy following liver cancer surgery, including gastric cancer, esophageal cancer, duodenal cancer, colon cancer and rectal cancer, which are cancers of the digestive system . Since the efficacy of the situation mushroom was found to be caused by polysaccharide beta glucan, research on polysaccharides has been actively conducted. In general, mushrooms contain potassium, calcium, magnesium, vitamins B2, B3, C, and fibrous amino acids. In addition to these ingredients, mushrooms contain polysaccharide β-glucan. Beta-glucan reinforces the body's immunity, repels bacteria and foreign substances that have invaded the body, and acts as a suppressor of infection. Phellinus sps (Berk) Pilat of the present invention belongs to the genus Phellinus ( Hymenochaetaceae ), the medical efficacy associated with the disease has long been well known in oriental medicine. It has anti-cancer effects, and the extract has also been proved by clinical trials as having an excellent therapeutic and preventive effect against liver cancer, lung cancer and many other cancers. However, the antiviral activity effect of the situation mushroom has not been studied.

The present invention was completed by confirming that the cleaning composition containing the combination composition of the situation mushroom extract and lactic acid according to the present invention had an excellent inhibitory effect against antibacterial and antiviral activity, in particular, HPV virus.

The present invention provides an effective and safe cleansing composition for preventing and treating intravaginal inflammation and cervical cancer containing the extract of the situation mushroom extract and lactic acid as an active ingredient.

In order to achieve the above object, the present invention provides a detergent composition containing lactic acid mushroom extract and lactic acid effective for preventing and treating intravaginal inflammation and cervical cancer.

The extract refers to an extract available in a polar solvent selected from water, lower alcohols having 1 to 4 carbon atoms, secondary alcohols or mixed solvents thereof.

The secondary alcohol includes ethylene glycol, propylene glycol, butylene glycol and the like.

The situation mushroom may be domestic, African, Cambodian, Chinese, preferably domestic.

The situation mushroom extract and lactic acid include a blending composition having a blending ratio of 1: 30 to 1, preferably a blending ratio of 1: 20 to 1, more preferably 1: 10 to 1.

Intravaginal inflammation includes viral and bacterial diseases.

The cleaning composition includes properties such as a liquid, a gel, and the like.

Hereinafter, the present invention will be described in detail.

The situation mushroom extract of the present invention is, after crushing and grinding the dried situation mushroom, about 1 to 10 times the weight (kg), preferably about 2 to 5 times the water, lower alcohol having 1 to 4 carbon atoms, Secondary alcohol or a mixed solvent of secondary alcohol and purified water having a mixing ratio (w / v) of about 1 to 10: 1, preferably 1 to 5: 1, more preferably of butylene glycol or propylene glycol and purified water Extraction method such as cold sediment extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, etc. for about 1 hour to 48 hours, preferably 10 hours to 30 hours at an extraction temperature of 0 to 100 ℃, preferably 10 to 30 ℃ as a mixed solvent A first step of obtaining the situation mushroom extract of the present invention is preferably repeated several times by cold extraction, followed by filtration with a pore size filtration membrane of 0.75-0.20 μm, preferably 0.30-0.20 μm;

1 g of commercially available lactic acid and 0.5 g of mushroom extracts are mixed with 100 mL of purified water, and a second step is obtained by adding other excipients to obtain genoclean, which is a cleaning composition of the present invention.

The present invention provides a cleansing composition which is obtained by the above method and is effective in the prevention and treatment of intravaginal inflammation and cervical cancer.

The cleaning composition for preventing and treating intravaginal inflammation and cervical cancer of the present invention comprises 0.1 to 80% by weight of the extract, preferably 1.0 to 50% by weight, based on the cleaning agent.

The combination composition of the situation mushroom extract and lactic acid according to the present invention was found to exhibit excellent bacterial growth inhibition and HPV-DNA expression inhibitory activity as a result of experiments on the inhibitory activity against bacteria causing vaginitis and HPV virus causing cervical cancer. It was.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1 Preparation of Situary Mushroom Extract

After washing 2kg of the situation mushroom purchased from Gyeongdong market, chop it up and immerse it in 8kg mixed solution of 1,3-butylene glycol or propylene glycol and purified water in a weight ratio of 2: 6 and soak for 10 days at room temperature. , Filtered by 0.22μm filtration membrane to prepare a situation mushroom extract of the present invention.

Example 2. Genoclean

1 g of commercially available lactic acid (Sigma, L6661) and 0.5 g of the situation mushroom extract prepared in Example 1 were added to 100 ml of purified water and mixed, followed by addition of a conventional cleaning composition to prepare a cleaning composition of the present invention. See example 1). The cleaning composition comprising the mixture of the situation mushroom extract and lactic acid of the present invention as an active ingredient was used as the sample G of the present invention (hereinafter referred to as ginoclean).

Reference Example 1. Preparation of HPV Infected Uterine Specimen

Cervical specimens were used after obtaining permission from H University Hospital's Obstetrics and Gynecology. The specimen is part of the specimen that was collected from the cervix of the patient and stored in the refrigerator for disposal after checking the HPV DNA amount and TYPE (16, 18, 33) by DIGEN KIT in the hospital. The inventors obtained some specimens for research purposes and transported them to BD's CELLMATICS virus transport medium to the laboratory, and once again, HPV DNA test was performed with DIGEN's KIT to confirm the presence of the virus and used in the experiment. Three different experiments were named No. 1, No. 2, and No. 3 for three experiments.

Experimental Example 1. Comparison of HPV-DNA inhibitory ability

In order to measure the inhibitory ability of the genoclean HPV virus against the DNA of the present invention, the DNA of the HPV virus present in the vaginal specimens was quantitatively tested by chemiluminescence on a microplate.

The experiment was performed by using RNA probes (RNA probes, ad & av stralms) capable of binding to DNA of HPV viruses contained in vaginal specimens, and then making DNA: RNA hybrids on microplates containing specific antibodies thereto. After capturing and binding the captured hybrid with an antibody conjugated with alkaline phosphatase, the chemiluminescent material is used to measure the amount of DNA of HPV virus present in the vaginal specimens by light intensity and the standard curve. As a method of determining the concentration through a calibration curve, a hybrid capture technology (Digene) was used for the experiment.

The experiment was measured with 20ul of three cervical specimens which confirmed the amount of HPV DNA in the specimen. After treating the genoclean containing the situation mushroom extract of Example 2 and reacting at 37 ℃ for 2 hours to compare the amount of HPV virus DNA of the specimen was confirmed the HPV virus DNA inhibitory effect of the genoclean of the present invention. The control group was divided into 10 μl treatment group (negative control group; N), 10 μl treatment group of lactic acid (L), and 10 μl treatment group of Acyclovir (positive control group; P).

First remove the microplate from the microplate heater and then immediately remove the sealer and capture the Calibrators, Controls (N and P) and Experimental Groups (G and L) respectively. 75 μl each was transferred to a microplate (microplate coated with anti-RNA: DNA hybrid antibody). The capture microplate was covered with a microplate sealer and incubated for 60 minutes at 1100 ± 100 rpm, 18-25 ° C. using a rotary stirrer. The filtrate of the capture microplate was removed using a pipette and then tapped on clean blotter paper 2-3 times to completely remove the filtrate. In a buffer containing sodium azide, 75 μl of a detection reagent combining RNA: DNA hybrid with an antibody conjugated with alkaline phosphatase was dispensed into each capture microplate well, and micro The plate sealer was used to cover the capture microplate wells and incubate at 20-25 ° C. for 30 minutes. Thereafter, the filtrate of the capture microplate well was completely removed, and then the capture microplate well was filled with the distillate washing solution including sodium azide, and then washed six times by turning it upside down. After the washed capture plate was left inverted for 10 to 15 minutes on a clean blotter paper to absorb the remaining filtrate, 75 μl of chemiluminescent material (CDP-Star ^™ , Chemilumlnescent substrate) was dispensed into each capture microplate well. The microplate sealer was used to cover the capture microplate wells and incubated for 15 minutes at room temperature under direct sunlight. Within 15 minutes after the reaction was completed, the amount of DNA of HPV virus present in the specimen was measured by light intensity using a DML 2000 Luminometer.

No.1 No.2 No.3 Negative Control (N) 28,448 7,771 64,692 Lactic Acid Single Treatment (L) 18,084 6,231 60,948 Zino Clean (G) 14,498 5,189 54,574 Positive control group (P) 14,752 7,089 56,770

 The results of treatment of each sample with cervical specimens confirming HPV (Human paphilloma virus) infection were measured using Digene HPV Hybrid Capture II.

As a result, as shown in Table 1, ginoclean containing a mixture of the situation mushroom extract (0.5g / 100ml) and lactic acid showed the highest inhibitory effect on the expression of HPV DNA, thus cleaning agents for preventing and treating cervical cancer It was confirmed that it can be usefully used as a composition.

Experimental Example 2 Bacteria Growth Inhibition Capability

In order to determine the growth inhibitory ability of the harmful bacteria of the genoclean of the present invention, the experiment was performed using the following colony measurement method.

Lactic acid 1g / 100ml + green mushroom 0.5g / 100ml (hereinafter referred to as G0.5) lactic acid 1g / 100ml + green mushroom 0.3g / 100ml (hereinafter referred to as G0.3), lactic acid 1g / 100ml + green mushroom 0.1 Four different concentrations of samples were prepared: g / 100ml (hereinafter referred to as G0.1) and lactic acid 1g / 100ml (hereinafter referred to as L). 1) Candida albcans , 2) Staphylococcus aureus , 3) E. coli The bacteria were cultured overnight at 37 ° C. in a certain amount (100ul), and each strain was diluted to a concentration of 10 ⁻⁵ cfu / ml to determine the number of bacteria. Each strain (100ul) and each test material (100ul) was mixed and reacted for 4 hours at 37 ℃, after the reaction was applied to 100ul each TSB agar medium. After incubation overnight at 37 ℃ incubator (Incubator), the results are shown in Figures 1 to 3 below.

As shown in Figures 1 to 3, even at low concentrations (lactic acid 1g / 100ml + 0.1g / 100ml lactic acid mushroom) was confirmed that the bacterial growth is markedly suppressed to exhibit excellent antimicrobial activity, thus the genoclean of the present invention as a cleaning composition It was confirmed that it can be usefully used.

The situation mushroom extract of the present invention can be administered in the following formulations, the following formulation examples are merely to illustrate the invention, whereby the content of the present invention is not limited.

Formulation Example 1. Genoclean solution (in 100 ml)

 Lactic Acid ... ....... 1g

 Situation Mushroom Extract ... ......... 0.5g

 Other Excipients

      Whey ... .......... 180mg

      ethanol................................................. .......... 1g

      Preservative (benzalkonium chloride, menthol) ..................................

Formulation Example 2 Ginoclean Gel (In 100 ml)

 Lactic Acid ... ...... 1g

 Situation Mushroom Extract ... ....... 0.5g

 Other Excipients

       Whey ... ........ 180mg

      ethanol................................................. .......... 1g

       Agar ... ........... 1g

    Preservatives (benzalkonium chloride, menthol) ..................... a very small amount

Genoclean containing the situation mushroom extract according to the present invention as an active ingredient to confirm that it exhibits excellent antiviral activity against antibacterial and antiviral activity, especially HPV virus, effective and safe cleaning composition for the prevention and treatment of intravaginal inflammation and cervical cancer Can be used as

Claims (4)

Cleansing composition for the prevention and treatment of intravaginal inflammation and cervical cancer, comprising a situation mushroom extract and lactic acid combination composition as an active ingredient. The cleaning composition according to claim 1, which is a liquid or a gel. The cleaning composition according to claim 1, wherein the situation mushroom extract and lactic acid have a mixing ratio of 1: 10 to 1 by weight. 2. The cleaning composition according to claim 1, wherein the situation mushroom extract is an extract soluble in a polar solvent selected from water, lower alcohols having 1 to 4 carbon atoms, secondary alcohols or mixed solvents thereof.

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