TWI360422B - - Google Patents

Description

丄JULKfZZ IX. Description of the invention: [Technical field to which the invention pertains] The present invention is in the form of a composition for applying to the feminine genitals of a milk-containing fermentation liquid. Under the female vagina will secrete a small amount of secretions, the knife secretion has the role of lubricating the vagina, which is indispensable for the vaginal system. However, in the case of poor hygiene or decreased resistance, there are sometimes cases where the secretions of the dangerous passages increase, the color changes, and the smell is odorous, that is, the sputum shows yin and inflammation. Abnormal secretions can cause extreme paralysis or pain in the affected part of the patient, which can lead to psychological and physical discomfort. The vaginitis caused by the _ is still possible to create a phenomenon of oblique yin and red phlegm is the vulvitis, vaginitis and vulvitis can be known as genital money. The causes of female genital infections are generally as follows: 1. The most common fungal infection of fungal infections is Candida albicans. Candida albicans is easy to grow in a warm and humid environment. When individual resistance is insufficient, it is easy to reverse infection. (such as lack of sleep, cold, before and after menstruation and during pregnancy): The symptoms after infection include itching, burning, genital red waist, milky blocky white 77' must, urethral pain, redness, urinary pain, etc. It is particularly uncomfortable before and after menstruation. 5 1360422 II. Bacterial Infections Common symptoms of bacterial infection include a large amount of grayish-white or yellow secretions after sexual intercourse, and a odor-like odor. Most patients do not have itching. Too frequent sexual intercourse or too frequent flushing of the vagina can cause an increase in the pH of the vagina, which is the main cause of bacterial infections, because the bacteria that cause infection are suitable for survival in an alkaline environment (pH > 4 5). Bacterial infections are the most common mixed-type anaerobic bacteria' bacteria causing bacterial infections including Escherichia (Escherichia (3), Pseudomonas aeruginosa (Pseud〇m〇nas age kiss "(4)), Staphylococcus aureus (Galta) .) and the hammering ball {Streptococcus). Third, the symptoms of Trichomonas infection Trichomonas infection is a secretion of pus yellow, similar to the secretion of bacterial infection, but trichomoniasis infection can also cause severe itching, bleeding after sexual intercourse, urinary pain and other phenomena, secretions have It will appear green foamy and smelly. It usually gets worse after menstruation. You can see the redness of the cervix on the epidermis during internal diagnosis. Most of the trichomoniasis infections are transmitted through sexual behavior. ^The normal vaginal environment is not sterile. It contains about six kinds of bacteria, including Staphylococcus, Escherichia coli, Diphtheria, Candida and Lactobacillus. The most abundant Lactobacilli are probiotics, while Staphylococcus, Escherichia coli, Diphtheria-like and Candida are bad bacteria, but when the amount of "bad bacteria" is small, the health of the vagina will not be bad. The effect, and as long as the vagina is maintained in the normal fungus phase, that is, the so-called "probiotics" predominate, the bad bacteria will not breed a lot, so there will be no such infections.

The If shape occurs. In addition, the value of the damage is about pH 3 8 4 9 , which is more secretive. If the _ 偏 4 4 = yummy of the road, there will be infection. Towards g// is more neutral or maintained at pH 3.8-4.2, and in this case, the pathogen of vaginal infection cannot grow, resulting in = and the current money towel & _ destruction, Infection status. There are currently methods for treating female genital infections, such as steroids and steroids (four), but this tends to ", and kills: so the initial treatment of the silk treatment is no = I, blood bacteria and bad bacteria Competition, but competition = long is often better than probiotics, so it is easy to occur. The second brother: the original brother:: repeatedly into the sputum, turn. But repeatedly make the occupational and steroid treatment, the pathogen produces resistance, and even cause the human body In addition, 'the vaginal skin care products currently on the market are mostly hard chemical agents, which can alleviate the symptoms of infection in the short term'. However, because the pH value is neutral, the vaginal pH value will increase after use and become suitable. Disease (4) The environment in which the disease grows, so the symptoms are easy to relapse, making the symptoms difficult to cure. Therefore, actively looking for a new vaginal infection-preventing composition that can inhibit the growth of pathogens while promoting the growth of probiotics without irritating the skin. SUMMARY OF THE INVENTION In order to solve the problems of the prior art described above, the object of the present invention is to provide a condition for maintaining normality in the vagina. A composition for use in a female genital area. Another object of the present invention is to provide a composition for the female genital area which maintains the vaginal acid test value at pH 3.8-4.2 so that the pathogen is not easily grown. A further object is to provide a composition for application to a female genital area which does not produce drug resistance. For the purpose of the present invention, a composition for a female genital area according to the present invention, comprising a lactic acid bacterium fermentation broth, The lactic acid bacterium fermentation liquid is obtained by inoculating a strain of Lactobacillus in a milk t and fermenting it at 30 to 45 ° C. Examples of the strain of the genus Lactobacillus include eosinophilic lactic acid. Bacillus (/^如办(10)·//(10)(10)), Lactobacillus bifidus, Lactobacillus brevis, Lactobacillus bulgaricus Aw/gflr/cws, Lactobacillus UuS casei ), Lactobacilhd S cellobiosus, QLactobdcilliis c〇Hin〇ides, Lactobacillus licheniformis i/e/Arwech·/, Lactobacillus fermentum (Lactobacillu) s fennentum), Lactobacillus JmseW, Lactobacillus gamma muscle, Lactobacillus plantamm, Lactobacillus rhamnosus, scented green scorpion {Lactobacillus, crimped Lactobacillus (ZflC/MflCZ·//(10) and Labradorella breve i/eArwech·/), preferably Lactobacillus acidophilus C^cio6acz7/(10)πζ·ί/ο;?/ζζ7(10)) and dried milk Bacillus strain 1360422 [Lactobacillus casef) The above strains of the genus Lactobacillus may be used alone or in combination with the strains of lactic acid rods of different species or different strains. There is no special restriction on the class. It can be used for the milk produced by various animals. It can also be a reduction gift. Examples of the milk produced by the animal include annual milk, preferably milk.

It can be easily understood by the skilled artisan that when the above fermentation culture is required, the fermentation culture temperature is determined according to the inoculum amount of the H branch (four) strain and the fermentation culture temperature, and the preferred fermentation culture time is 12 ~72 hours 'better for 12 to 36 hours. The above-mentioned fermentation bacterium fermentation broth can be further treated by the step of removing the bacteria, and the step of removing the bacteria is not limited, and the separation includes centrifugation and sputum. The composition of the present invention can be used to promote the growth of probiotic bacteria in the vagina

This maintains the pH in the vagina within the normal range (pH 3.8-4.2), since conventional pathogens are not susceptible to growth at this pH. Therefore, the composition of the present invention can prevent the pathogenicity of a pathogen (i.e., a microorganism causing vaginal infection) to maintain the bacteriological phase in the vagina in a normal state, and thus can be used for preventing and treating female genital infection. The above pathogen may be any microorganism which causes vaginal infection, and examples thereof include trichomoniasis, mold g and bacteria, wherein the bacteria are mainly a large intestine function ('chia c〇li), a green bacterium (pseud 〇) It is similar to ae "Uginosa", Staphylococcus aureus (汾印知(10)) and Streptococcus 9 1360422 (Streptococcus). In addition to the above characteristics, the composition of the present invention is mild and non-irritating, and is not used for a long time. It can cause skin irritation or irritation, so it can be used for the cleaning and dressing of female genitals on a regular basis and after the financial period. In addition, the invented composition can quickly reduce the PH# of the vagina, avoiding the increase in the pH of the vagina after sex, so it is available After female sexual behavior, vagina ^ clean 0

In order to facilitate the present invention, the present invention can be prepared into various suitable dosage forms as needed, and examples thereof include sprays, gels, gels, lotions, creams, creams, and stoppers. Agents, powders, paper towels and cotton swabs, but are not limited to this.

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. It will be apparent to those skilled in the art that the present invention may be modified and modified without departing from the spirit and scope of the invention, and the scope of the present invention is defined by the scope of the appended claims. [Embodiment] Example 1 Lactic acid bacteria fermentation broth, Lactobacillus acidophi (4) and Lactobacillus faecalis (Lact〇bacUlus, numbered BCRC 14079 and BCRC 10697, respectively) were purchased from the Food Industry Development Institute. Two strains of I strain were simultaneously inoculated into fresh milk. The two strains were selected from natural lactic acid bacteria in the physiological urinary system of healthy women, and then cultured for 24 hours to obtain a lactic acid bacteria fermentation broth. The fermentation broth is separated from the ~' solid solution, and the supernatant is taken and filtered by the membrane to remove the insoluble process contained in the fermentation broth in a clean room of U), 000 class.

In this example, four strains were used, namely Escherichia coli <^), Pseudomonas aeruginosa (housefly muscle y, Staphylococcus aureus (9) Ink/Qingsheng (10)) and Streptococcus (9) The strains were all taken from the clinical center of the Medical Center of Zhongshan Medical University and confirmed by the standard operating procedures of the bacteria room of the laboratory of the medical center. The aforementioned four bacteria were inoculated separately into 5 mL of Luria-Bertani medium and at 37. (: The culture was shaken at 200 rpm. After 16 hours of culture, the 0.5 mL cultured bacterial solution was added to the fermentation broth containing 1 mL, 0.8 mL, 0.6 mL, 0.4 mL, 0.2 mL, and 0 mL of the lactic acid bacteria of the present invention. In mL Luria-Bertani medium, incubate at 200 C for 4.5 hours at 37 ° C. The concentration of the bacteria solution (〇D600 value) was measured with a spectrometer set at 6 〇〇 nm. The fermentation broth containing 0 mL of the lactic acid bacteria of the present invention was used. The concentration of the bacterial liquid was taken as 100% growth, and the fermentation of 0.2 H1L lactic acid bacteria was calculated, 0.8 mL, 0.6 mL, and 0.4 mL. Invented lactic acid _ liquid is not inhibited = strain::: The hair in the lower pain is stored white + 'visible To the above lactic acid bacteria fermentation broth has two W for four bacteria. / Female effect curve 'The lactic acid bacteria fermentation broth is 50 for Staphylococcus. Pi Chen degree) is about 15%, Example 3 for Escherichia coli and Pseudomonas aeruginosa; lLil reading fermentation broth 夭 夭 夭 的 皮肤 皮肤 皮肤 皮肤 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本 本The dose of the agent is produced after the induction period, and then an active dose of the agent is administered to observe whether an allergic reaction occurs, and this is an attack period. In this example, the skin reaction is divided into no reaction: 0, slight erythema: 1, moderate erythema: 2, intensity redness and edema (severe erythema and edema): 3 . At the beginning of the experiment, the guinea pigs were divided into the control group (reverse osmosis deionized water) and the positive control group (2, 4-dinitro- 4 chlorobenzene (2, 12:8) 1360422 4-dinitnM-Chl〇robenzene, DNCB )) and the lactic acid bacteria fermentation broth group lactic acid bacteria fermentation broth), the positive control group in the induction period and the attack period root i Shirasu and Matsuoka test concentration, respectively, using mL 5 mL. Undiluted lactic acid bacteria in lactic acid bacteria fermentation broth

The l% (w/v) DNCB and 0.5 mL 〇.1% (w/v) dnCB groups were treated with 〇 5 mL fermentation broth during the induction and challenge phases. In the present example, the control group includes 10 guinea pigs, five of which are male and five are female; the positive control group includes one guinea pig, five of which are male and five are female; the lactic acid fermentation broth includes 2 竺 竺 ,, 10 of which are male, + only _ mouse. In the first three weeks, the hairs of the 4x6 Cm region of the right scorpion of the three groups of guinea pigs were shaved, and 5 mL of reverse osmosis deionized water, 0.5 mL of 1% DNCB, and 0.5 mL of the lactic acid bacteria fermentation broth of the present invention were separately coated. One square inch of four layers of gauze, respectively, attached to each group of guinea pigs in the right waist shaving area 'and then breathable tape (Micr〇p〇reTM, 3M, 81,] \ ^, 1; 8 eight) and adhesive elasticity Bandage (£; 1^7^, 8 1 (^, 1^0, Japan) for six hours' after six hours, remove the gauze, reverse the osmotic deionized water, and remove the gauze 24 and 48 hours after removing the gauze. The redness of the shaved area was observed and recorded. The fourth day of the day, the mole was not treated with the drug. On the fifth week, the hair of the left flank of each group of guinea pigs was shaved off one day before the test, and 5 mL of 10% was taken. Sodium sulfate (SDS) (w/v) (Osaka and Koko Pure Chemical Industries, Ltd.) was applied to the shaved area to enhance skin irritation and allergies. After six hours, the reverse osmosis deionized water was used to clean the remedy. Take 0.5 mL of reverse osmosis deionized water, 〇 5 mL of 〇.i〇/0 DNCB and 0.5 mL of this 1360422 fermentation every other day. They were coated on the four-layer gauze of - square inch respectively. 'The strong left hair area of the scorpion scorpion of each group, and then the breathable tape and the bag purple for twenty-four hours. After 24 hours, remove the gauze. (4) L ion The water is flushed with the medicine, 24, 48 and 72 after the gauze is removed, and the redness of the shaved area is recorded. The results from the first week to the sixth week are not shown in Tables 2 to 4. The liquid group had no skin from the first week to the third week. Referring to Tables 2 to 4, it can be seen that the guinea pigs in the positive control group showed moderate redness in the right lower lumbar skin for two weeks, and the second and the week appeared. Symptoms of moderate to intense redness and swelling, and secretory skin production. „The acid bacteria fermentation liquid group had no skin redness and anti-U skin allergy positive rate in the fifth and sixth weeks, and the positive control group’s guinea pig was in 2 The sixth test showed that the redness of the towel was too difficult, and the money was allergic. The 1〇〇/0 'grade belongs to ν, which is “extremely allergic drug.” The positive rate of skin allergy to Zhao group and lactic acid bacteria fermentation liquid group is Q%. , the grade belongs to, !,, is "no allergic". Therefore, it is available It is known that the lactic acid bacteria fermentation broth of the present invention is non-dermal sensitization. In addition, the body weight of guinea pigs is measured and recorded in the first to sixth weeks, and the results are shown in Tables 5 and 6. The results of the observations show that the positive sputum is positive. In the fifth week of the male rats in the group, the females showed a significant downward trend (P<0.05) in the sixth material, and the weight was measured by the Duncan multiple range test. However, there was no significant difference between the γ lactic acid fermentation broth group and the control group, and it was found that the lactic acid bacteria fermentation broth did not affect the body adversely. θ at 14 1360422 Example 4 Skin allergy test of the lyophilized white strip of the yeast solution This example includes 6 white rabbits, and the normal area of the back skin is selected, and the back of the white rabbit is about 4x6 cm area by electric shaving scissors one day before the test. Hair shaving can't hurt the skin when shaving. Next, take 0.5 mL of the pulverized lactic acid fermentation broth and place it on a square inch of gauze wetted with distilled water. Place the gauze on the shaved area on the back of the white rabbit, and then use a breathable tape (MicroporeTM, 3M, St , MN, USA) and adhesive elastic bandage (ELATEX, Alcare, Tokyo, Japan) were bandaged for four hours. After four hours, the gauze was removed and the drug was flushed with sufficient water and observed at 4, 24, 48 and 72 hours. Also record the irritation of the skin. If the skin irritation does not disappear, continue to observe for 4, 7 and 14 days. According to the criteria of the Draize method, the stimulation index (redness/crustosis and edema) was added, and then the individual white rabbit skin irritation index (Primary, irritation index) was obtained. The average index value obtained is the skin irritation index level of the lactic acid bacteria fermentation broth to white rabbits. The results are shown in Table 7. It can be seen from Table 7 that the skin area treated with the lactic acid bacteria fermentation broth has no obvious redness after removal of the gauze. According to the evaluation method such as skin irritation index, the average susceptibility index of redness, journey and water in the lactic acid bacteria fermentation broth at 4, 24, 48 and 72 hours was G. According to the skin irritation index evaluation method of the plus test, the milk (four) fermentation broth has no skin irritation to the skin irritation grade of the white rabbit. v 15 1360422 Table 1 Survival rate of each strain in the presence of different lactic acid bacteria fermentation broth (%) Lactic acid bacteria fermentation broth (mL) Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus 1.0 21.0 32.4 47.1 5.4 0.8 36.9 51.6 57.9 16.7 0.6 71.0 57.1 75.4 49.7 0.4 88.0 93.6 88.9 73.7 0.2 100.0 94.2 94.5 100.0 0.0 100.0 100.0 100.0 100.0

Table 2: Response of the skin of the positive control group during the induction period of the guinea skin allergy test

First week second week third week gender number 24 hours 48 hours 24 hours 48 hours 24 hours 48 hours male rats 311 1 1 4 3 4 4 312 1 1 3 2 3 2 313 1 1 4 3 3 2 314 1 1 4 4 3 3 315 1 1 3 4 4 4 Female 316 1 1 3 3 3 2 317 1 1 3 3 3 2 318 1 1 4 3 2 3 319 1 1 4 4 4 3 320 1 1 4 4 3 3 16 1360422 Response of the skin of the positive control group during the attack period of the three-day squirrel skin allergy test. Week 5 Week 6 Gender number 24 hours 48 hours 24 hours 48 hours Male 311 1 0 4 4 312 1 1 1 1 313 1 0 1 1 314 1 1 4 4 315 1 0 4 4 Female 316 1 0 4 4 317 1 1 4 4 318 1 0 1 1 319 1 0 4 4 320 1 0 4 4

Table 4 The level and classification of the skin allergy test of the mole rat

Sensitivity of all positive animals in all test animals (%) Graded control group 100 (0/10) I No allergic positive control group 10 10 100 (10/10) V Extremely allergic lactic acid bacteria fermentation broth group 20 0 0 (0/20) I No allergies 17 1360422

The weight change of male rats in the skin sensitization test of the fifth day of the first week, the second week of the third week, the fourth week, the fourth week, the fifth week, the sixth week, the average value of the control group 327.0 354.8 403.4 474.5 497.4 517.1 standard deviation 7.1 9.6 15.5 20.2 19.0 32.8 positive control Mean 329.2 378.4 401.6 458.9 365.1 417.5 Group standard deviation 23.3 19.7 20.7 20.6 34.4 24.0 Average value of lactic acid bacteria 349.4 365.9 381.0 438.3 467.7 503.3 Standard deviation of the yeast solution group 16.8 24.3 31.2 31.3 27.9 30.5 Weight changes of male rats in the skin allergy test of the six-day squirrel First week, second week, third week, fourth week, fifth week, sixth week, sixth week, control group average 324.2 351.8 393.4 421.9 440.8 487.7 standard deviation 14.6 33.6 29.1 34.6 37.6 43.7 positive control average 322.4 . 344.2 371.6 396.0 441.0 425.6 standard deviation 13.0 13.6 20.3 34.8 27.3 35.8 Average value of lactic acid bacteria 332.0 345.4 368.2 427.1 464.8 Standard deviation of yeast solution group 19.5 20.1 23.9 30.1 15.3

18 Table 7 Skin irritation index of white rabbit skin allergy test 4 hours 24 hours 48 hours 72 hours redness / knot 0.0 0.0 0.0 0.0 Edema 0.0 0.0 0.0 0.0 Average index 0.0 0.0 0.0 0.0 1360422 [Simple diagram] No [main components Explanation of symbols] No 19

Claims (1)

1360422 丨.- Crane for lion and / or treatment of female genitals (four) of the medical and health care composition, comprising - lactic acid bacteria fermentation broth by lactic acid bacteria by LacGbaeillus aeid GphiIus and Lactobacillus - The Lactebacmus casei) strain is inoculated into a milk, and is obtained by fermentation and removal of the cells under 3〇~机鲁, the lactic acid bacteria fermentation broth is non-dermal sensitizatiGn' and can promote probiotics in the vagina. Growing. 2. For the (10) health care composition as described in the special item, the lactobacillus fermentation broth is between 30 and 45. (: The fermentation is carried out for 12 to 72 hours. 3. For the application of the second embodiment of the medicinal composition, the lactic acid fermentation broth is fermented for 12 to 36 hours at 30 to 451. The medical and health care composition according to claim 1, wherein the step of removing the cells is performed by centrifugation. 5. The medical and health care composition according to claim 1, wherein the removal is The step of the cell is filtered. 6. The medical refractory composition according to claim 1, wherein the female genital infection is caused by a pathogen. 7. The medical and health care composition according to claim 6 Wherein the disease is 20 1360422. The original system is selected from the group consisting of trichomoniasis, mold and bacteria. 8. The pharmaceutical health care composition according to claim 7 wherein the bacterium is selected from the group consisting of Escherichia coli ( Escherichia coli), a group consisting of Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus 9. The pharmaceutical health care composition of claim 1 Wherein the composition is a dosage form selected from the group spray, condensation, gel, lotion, cream, ointment, suppository, powder, paper towels and swabs thereof. twenty one