Traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention provides a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects, which is characterized by comprising a fermentation product formed by fermenting a Chinese herbal medicine preparation and compound probiotics, and the Chinese herbal medicine preparation comprises the following raw material medicines: cape jasmine, saussurea involucrata, hairyvein agrimony, radix stemonae, fructus cnidii and golden cypress. The invention further provides a preparation method of the traditional Chinese medicine composition with the vaginal antibacterial and anti-inflammatory effects and application of the traditional Chinese medicine composition in preparation of products for preventing or treating vaginal antibacterial and anti-inflammatory effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of female care, and specifically relates to a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects and a preparation method thereof, and also relates to the application of the composition in the preparation of products for preventing or treating vaginal antibacterial and anti-inflammatory effects. Background Art

[0002] Vaginitis is a common gynecological disease, often manifested by vulvar itching, local erosion, increased vaginal discharge, accompanied by symptoms such as frequent urination and dysuria. It is also one of the gynecological diseases with a higher incidence rate. If not treated in time, it can easily cause cervicitis. In severe cases, it can infect the pelvic cavity, upper reproductive tract, abdominal cavity, etc. It is a common and frequently occurring gynecological disease with a high recurrence rate and is distributed in all age groups, affecting the patient's physical and mental health and quality of life.

[0003] In order to prevent the occurrence of vaginitis and relieve the symptoms of vaginitis, many women use various gynecological cleansers to clean their private parts. There are two types of female private part care products currently on the market. One type uses chemical bactericides such as chlorhexidine acetate. The skin of the female private parts is relatively sensitive. A large amount of chemical bactericides can easily cause skin irritation. Long-term use can also easily lead to bacterial resistance and destroy beneficial bacteria. The other type is Chinese herbal medicine products, and the dosage forms and usages are mostly flushing agents or suppositories. Since natural Chinese herbal medicine ingredients are used, they are less harmful to the human body, have no side effects, and are safe and reliable. However, for example, Chinese patent document CN1939518A discloses a herbal care solution for women's menstrual period and daily use that shrinks the vagina, kills bacteria and relieves itching. Since the natural Chinese herbal medicine ingredients used in the patented technology are all water extracts, the effective ingredients extracted from each Chinese herbal medicine ingredient are very limited. In order to increase the content of the effective ingredients, natural water such as well water is used for soaking and extraction, and the preparation process is more demanding. Patent CN117503838A discloses a probiotic compound preparation, including a matrix and an additive, wherein the matrix includes Lactobacillus crispatus, Lactobacillus acidophilus and Bifidobacterium, and the additive includes Phellodendron chinense, Sophora flavescens, Salvia miltiorrhiza, Gardenia jasminoides, Psoralea corylifolia and Licorice. The probiotic preparation provided by the patent is a live bacteria preparation, which can be used for regulating vaginal flora and inhibiting Gardnerella. However, live bacteria preparations have many restrictions in use and storage, and regulations do not allow live bacteria preparations to be used in daily chemical products.

[0004] Therefore, it is very necessary to develop a female private parts care product that has mild antibacterial and anti-inflammatory effects, a simple and easy-to-control preparation method, and a probiotic fermentation raw material that can be used in daily chemical products. Summary of the invention

[0005] In view of the defects of the prior art, the purpose of the present invention is to provide a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects and a preparation method thereof, as well as an application of the composition in the preparation of products for preventing or treating vaginal antibacterial and anti-inflammatory effects.

[0006] To achieve the above purpose, the solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects, comprising a fermentation product formed by fermenting a Chinese herbal medicine preparation and a composite probiotic, wherein the raw materials of the Chinese herbal medicine preparation include gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron.

[0008] Preferably, the Chinese herbal medicine preparation is the powder of each raw material medicine, or the water or organic solvent extract of each raw material medicine, preferably the powder of each raw material medicine; the composite probiotics include a first probiotic and a second probiotic, the first probiotic is selected from one or more of lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), yeast fungi (Saccharomyces cerevisiae, Pichia pastoris), preferably Bacillus subtilis; the second probiotic is selected from one or more of Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii and Lactobacillus gasseri, preferably Lactobacillus crispatus.

[0009] Preferably, in the raw materials of the Chinese herbal preparation, the mass ratio of Gardenia, Saussurea involucrata, Herba Agrimoniae, Radix Stemonae, Fructus Cnidii and Phellodendron amurense is (1-3): (1-3): (1-5): 1:1:1, and the particle size of the powder of each raw material of the Chinese herbal preparation is 40-200 mesh; the bacterial count ratio of the first probiotic and the second probiotic is (1-5): 1.

[0010] In a second aspect, the present invention also provides a method for preparing the above-mentioned traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects, comprising the following steps:

[0011] Step (1), weighing gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron chinense according to the proportion, crushing, and passing through an 80-mesh sieve to obtain a mixed powder of Chinese herbal medicines;

[0012] Step (2), weighing a certain amount of mixed powder of Chinese herbal medicines, adding 5-10 times of water, stirring, and sterilizing at 121° C.;

[0013] Step (3), after cooling, inoculating the composite probiotic strain in an ultra-clean workbench and fermenting at 25°C-30°C for 3d-7d;

[0014] Step (4), sterilize after the fermentation is completed, dry the fermented medicinal materials in a constant temperature oven at 60°C to constant weight, add 15 times of 50% ethanol and extract at 75°C for 1 hour. After the extraction is completed, filter, add 10 times of 50% ethanol to the residue and extract at 75°C for 1 hour. After the extraction is completed, filter, combine the two filtrates, filter using a 0.22μm filter membrane, decompress and concentrate the filtrate, and then freeze-dry to obtain a dry fermented plant extract.

[0015] Preferably, the composite probiotics include a first probiotic and a second probiotic, wherein the first probiotic is selected from one or more of lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), and yeast fungi (Saccharomyces cerevisiae, Pichia pastoris), preferably Bacillus subtilis; and the second probiotic is selected from one or more of Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii and Lactobacillus gasseri, preferably Lactobacillus crispatus.

[0016] Preferably, for every 100 g of the mixed powder of Chinese herbal medicine, the volume of the inoculated probiotic seed solution is 10-20 mL.

[0017] In a third aspect, the present invention also provides a use of the above-mentioned traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects in the preparation of a product for preventing or treating vaginal antibacterial and anti-inflammatory effects.

[0018] In a fourth aspect, the present invention further provides an external preparation having vaginal antibacterial and anti-inflammatory effects, including the traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects as described above.

[0019] Preferably, the dosage form of the external preparation with vaginal antibacterial and anti-inflammatory effects includes ointment, spray, douche or suppository, preferably ointment.

[0020] Preferably, the external preparation having vaginal antibacterial and anti-inflammatory effects further comprises at least one of an excipient, a bactericide, a moisturizer, an anti-allergic agent, a chelating agent and a flavor.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The pharmaceutical composition with vaginal antibacterial and anti-inflammatory effects prepared by the present invention combines several Chinese herbal medicines with microbial fermentation technology, and degrades the effective active ingredients in the Chinese herbal medicines that cannot be directly utilized after entering the human body into small-molecule active substances through microbial fermentation, thereby greatly improving the efficacy of the several Chinese herbal medicines, and the small-molecule active ingredients in the fermentation products have a strong transdermal permeability, thereby significantly improving the vaginal antibacterial and anti-inflammatory effects; at the same time, because the pharmaceutical composition adopts the probiotics Bacillus subtilis and Lactobacillus crispatus for fermentation, it has little stimulation to the vagina, and long-term use will not destroy the balance of the vaginal flora itself. DETAILED DESCRIPTION

[0023] The invention provides a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects, comprising a fermentation product formed by fermenting a Chinese herbal medicine preparation and a composite probiotic. The raw materials of the Chinese herbal medicine preparation include gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron.

[0024] In the traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects provided by the present invention, the Chinese herbal medicine preparations used can be powders of various raw materials, or can be water or organic solvent extracts of various raw materials.

[0025] In a Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects provided by the present invention, the composite probiotics used include a first probiotic and a second probiotic. The first probiotic includes but is not limited to lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), yeast fungi (Saccharomyces cerevisiae, Pichia pastoris), and in the embodiments of the present invention, Bacillus subtilis is used as an example to illustrate the technical solution of the present invention. The second probiotic includes but is not limited to Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus gasseri, etc., and in the embodiments of the present invention, Lactobacillus crispatus is used as an example to illustrate the technical solution of the present invention.

[0026] The present invention adopts, for example, Bacillus subtilis as the first probiotic, which has a very rich enzyme system, has a powerful ability to decompose and transform substances, and ferments Chinese medicine with probiotic microorganisms, can carry out biotransformation on the chemical components of Chinese medicine, produce new compounds or cause the changes of some component contents in Chinese medicine, and some components in Chinese medicine may also induce certain metabolic pathways of probiotic microorganisms to change, thereby producing new compounds. It has been proved through research experiments that probiotic microorganisms can grow in most Chinese medicines, and Chinese medicines can provide the necessary nutritional conditions and physiological regulatory factors for microbial growth. Further research has also found that the cellulase produced by Bacillus subtilis in the growth and reproduction process can destroy the cell wall of Chinese medicine, so that its effective ingredients are fully released, thereby greatly improving the efficacy of Chinese medicine, except producing the above-mentioned cellulase, it can also produce more than a dozen enzymes such as protease, amylase, phytase, pectinase, etc., which determines that when Bacillus subtilis is fermenting Chinese medicine, multiple enzymes can ferment the corresponding substrate in Chinese medicine, which will improve the efficacy of Chinese medicine. In addition, Bacillus subtilis also produces active substances such as subtilisin and gramicidin during its growth and reproduction, which have a significant inhibitory effect on pathogenic bacteria. Bacillus subtilis also rapidly consumes free oxygen during its growth and reproduction, promotes the growth of beneficial anaerobic bacteria, maintains and regulates the balance of microecology, enhances cellular immunity and humoral immunity, and indirectly inhibits the growth of pathogenic bacteria, thereby greatly improving the body's disease resistance.

[0027] The present invention uses Lactobacillus crispatus as the second probiotic. It has been reported that 71.3% of the vaginal flora are Lactobacillus, of which Lactobacillus crispatus has the highest proportion, accounting for 37.0%, Lactobacillus iners accounts for 26.1%, Lactobacillus jensenii accounts for 4.23%, and Lactobacillus gasseri accounts for 4.23%. Studies have shown that the present invention uses Lactobacillus crispatus, which has the beneficial effects of regulating human vaginal flora, inhibiting pathogenic bacteria, and improving body immunity.

[0028] The embodiments of the present invention prove that the antibacterial and anti-inflammatory effects of the Chinese herbal medicine fermentation of Gardenia, Saussurea involucrata, Herba Agrimoniae, Radix Stemonae, Fructus Cnidii and Cortex Phellodendri on the vagina are not obvious. However, the use of Bacillus subtilis alone has a significant inhibitory effect on the antibacterial and anti-inflammatory effects of the vagina. More importantly, after the combination of Lactobacillus crispatus and Bacillus subtilis, the antibacterial and anti-inflammatory effects on the vagina are significantly improved, exceeding the effect of any one of the bacteria acting alone, so that the dosage of the Chinese herbal medicine can be reduced while achieving the same antibacterial and anti-inflammatory effects, reducing the potential risks of high-dose application.

[0029] In the traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects provided by the present invention, the fermentation product can also be extracted with water or an organic solvent. The organic solvent is, for example, ethanol, ethyl acetate, chloroform, acetone or petroleum ether. In the embodiment of the present invention, ethanol is used as the organic solvent for extraction.

[0030] In a traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects provided by the present invention, in the raw materials of the Chinese herbal preparation, the mass ratio of Gardenia jasminoides, Saussurea involucrata, Herba Agrimoniae, Radix Stemonae, Fructus Cnidii and Phellodendron chinense is (1-3): (1-3): (1-5): 1:1:1, for example, 2:2:3:1:1:1, and the particle size of the powder of each raw material of the Chinese herbal preparation is 40-200 mesh; the bacterial number ratio of the first probiotic and the second probiotic is (1-5): 1, for example, 2:1, 3:1, 4:1.

[0031] Furthermore, the present invention also provides a method for preparing the above-mentioned traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects, comprising the following steps:

[0032] Step (1), weighing gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron chinense according to the proportion, crushing, and passing through an 80-mesh sieve to obtain a mixed powder of Chinese herbal medicines;

[0033] In this step, the mixed powder of Chinese herbal medicine is obtained by grinding the raw materials of Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri and Phellodendron amurense, and then passing through an 80-mesh sieve and mixing them evenly.

[0034] Step (2), weighing a certain amount of mixed powder of Chinese herbal medicines, adding 5-10 times of water or organic solvent, soaking and sterilizing at 121° C. for 15-30 minutes;

[0035] Step (3), after cooling, inoculating the composite probiotic strain in an ultra-clean workbench and fermenting at 25°C-30°C for 3d-7d;

[0036] In this step, as mentioned above, the composite probiotics include a first probiotic and a second probiotic, wherein the first probiotic includes but is not limited to lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus, Streptococcus faecalis), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), yeast fungi (Saccharomyces cerevisiae, petroleum yeast), and in the embodiment of the present invention, Bacillus subtilis is used as an example to illustrate the technical solution of the present invention. The second probiotic includes but is not limited to Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus gasseri, etc., and in the embodiment of the present invention, Lactobacillus crispatus is used as an example to illustrate the technical solution of the present invention.

[0037] In this step, for every 100g of the mixed powder of Chinese herbal medicine, the volume of the inoculated composite probiotic seed solution is 10-30mL, so that the bacterial number ratio of the inoculated first probiotic and the second probiotic is (1-5):1, for example, 2:1, 3:1, 4:1. Step (4), after the fermentation is completed, sterilization is performed, the fermented medicinal material is dried in a 60°C constant temperature oven to constant weight, 15 times of 50% ethanol is added to extract at 75°C for 1h, after the extraction is completed, filtration is performed, and the filter residue is further added with 10 times of 50% ethanol to extract at 75°C for 1h, after the extraction is completed, filtration is performed, the two filtrates are combined, and filtered using a 0.22μm filter membrane, and the filtrate is concentrated under reduced pressure and freeze-dried to obtain a dry fermented plant extract.

[0038] The present invention also provides a product for preventing or treating vaginal antibacterial and anti-inflammatory diseases, comprising the traditional Chinese medicine composition.

[0039] In the present invention, the "Chinese medicine composition" and "product for preventing or treating vaginal antibacterial and anti-inflammatory" are generally compositions comprising a composition having active ingredient components (referred to as active ingredient) and pharmaceutically acceptable excipients.

[0040] In the present invention, the active ingredient components are fermentation products formed by fermenting Chinese herbal medicine preparations with probiotics, and the raw materials of the Chinese herbal medicine preparations include Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri and Phellodendron amurense.

[0041] In the present invention, the "pharmaceutically acceptable excipients" include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or lubricating oils.

[0042] The present invention provides a product for preventing or treating vaginal antibacterial and anti-inflammatory diseases, comprising the above-mentioned Chinese medicine composition, wherein female subjects for preventing or treating vaginal antibacterial and anti-inflammatory diseases show symptoms of dysbiosis of microbiota in the urogenital tract, such as bacterial vaginosis, candidal infection, human papillomavirus infection, urinary tract infection, sexually transmitted infection, etc. The product for preventing or treating vaginal antibacterial and anti-inflammatory diseases, comprising the above-mentioned Chinese medicine composition, provided by the present invention, is formulated for vaginal application, and its dosage form is an ointment, spray, douche or suppository, which can be achieved by adding at least one of an excipient, a bactericide, a moisturizer, an anti-allergic agent, a chelating agent and a fragrance to the composition, to prepare a vaginal external preparation in the form of a douche, and when applied, the douche is applied to the vaginal cavity through a tool such as an applicator.

[0043] In the traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects provided by the present invention, each raw material component as an active ingredient has the following expected functions:

[0044] Gardenia, also known as wood dan, fresh branch, Yuetao, branch, is the dried mature fruit of Gardenia Cardeniajasminoides Ellis, a plant of the Rubiaceae family. It has the effects of purging fire and relieving restlessness, clearing heat and removing dampness, cooling blood and detoxifying. It is mainly used to treat fever and restlessness, damp-heat jaundice, bloody stranguria and pain, blood-heat vomiting and bleeding, red and swollen eyes, and fire poison sores. Gardenia has an inhibitory effect on Staphylococcus aureus, meningococcus, catarrhalis, etc. Gardenia water extract has an inhibitory effect on a variety of skin fungi in vitro. The decoction has the effect of killing Leptospira and Schistosoma japonicum adults. Gardenia alcohol extract has a sedative effect but no analgesic effect. Gardenia extract has a certain anti-inflammatory and soft tissue injury treatment effect.

[0045] Snow lotus, also known as snow lotus, is the whole herb of Xinjiang snow lotus Saussurea involucrata Kar.etKir.ex Maxim., a plant of the genus Saussurea of ​​the Asteraceae family. It has the effects of promoting blood circulation and menstruation, dispelling cold and removing dampness, strengthening tendons and assisting yang. It is mainly used to treat rheumatoid arthritis, lung cold cough, lower abdominal cold pain, amenorrhea, retained placenta, and impotence. Snow lotus has the effects of warming the kidney and strengthening yang, dispelling wind and removing dampness, relieving pain, regulating menstruation and stopping bleeding. It can assist in the treatment of rheumatic arthralgia, lung cold cough, female lower abdominal cold pain, irregular menstruation, metrorrhagia, retained placenta, and impotence. Snow lotus has significant anti-rheumatic, analgesic, and anti-inflammatory effects. It has a good effect on relieving acute attacks of gout. Long-term use can also reduce the number of attacks without obvious toxic and side effects. It has anti-tumor effects, can inhibit the growth of cancer cells, and can assist in the treatment of cancers such as breast cancer, skin cancer, leukemia, and Hodgkin's disease. In addition, snow lotus can scavenge free radicals, reduce oxygen consumption, and achieve anti-fatigue effects; it has anti-radiation functions and can effectively prevent ultraviolet damage to the skin; it can also relieve smooth muscle spasms and can assist in the treatment of gastric ulcers.

[0046] Agrimonia, also known as crane grass bud, dragon tooth grass, etc. It tastes bitter and astringent, and is neutral in nature; it enters the heart and liver meridians. Agrimonia is the dried aerial part of Agrimonia pilosa Ledb, a plant of the genus Agrimonia in the Rosaceae family. It has the effects of astringent hemostasis, antidysentery, antimalarial, and tonic. It is mainly used to treat various bleeding syndromes, chronic diarrhea and dysentery, cold and heat malaria, qi and blood deficiency, exhaustion, etc. Agrimonia alcohol extract can contract peripheral blood vessels and has a significant procoagulant effect. Agrimonia phenol has a definite inhibitory and killing effect on Taenia solium, cysticercosis, larvae, Taenia solium and short shell tapeworms, and has an inhibitory and killing effect on Plasmodium and Trichomonas vaginalis. Agrimonia also has antibacterial and anti-inflammatory, anti-tumor, and analgesic effects.

[0047] Stemona, also known as: Stemona root, Baibing, Yuxiao, etc. It tastes sweet and bitter, is slightly warm in nature, and enters the lung meridian. Stemona is the dried root of Stemona ja ponica (BL.) Miq., Stemona sessile folia (Miq) Franch. et Sav., or Stemona tuberosa Lour., etc., of the Stemonaceae family. It has the effects of moistening the lungs and relieving coughs, killing insects and lice. It is mainly used to treat new and long-term coughs, whooping coughs, coughs caused by tuberculosis, pinworms, vaginal trichomonas, head lice and scabies. Stemona contains a variety of alkaloids, such as stemonine, stemonidine, prostemonine, substemonine, erect stemonine, parastemonine, trepanning stemonine, etc., as well as sugars, lipids, proteins, succinic acid, etc. The decoction and wine extract of Stemona have different degrees of inhibitory effect on pneumococcus, beta-hemolytic streptococcus, meningococcus, Staphylococcus aureus, Staphylococcus albus, Shigella dysenteriae, Typhoid Bacteria, Paratyphoid Bacteria, Escherichia coli, Proteus, Diphtheria Bacteria, Plague Bacteria, Subtilis Bacteria, Vibrio cholerae, and Tuberculosis Bacteria; it has a certain inhibitory effect on certain viruses and pathogenic skin fungi. The water or alcohol extract of Stemona has the effect of expelling and killing head lice, clothing lice, pubic lice, bedbugs, fly maggots, pinworms, etc., and has a hypotensive effect, and can also dilate coronary blood vessels and inhibit myocardial contractility.

[0048] Cnidium monnieri, also known as snake rice, snake pearl, snake millet, cnidium kernel, etc., tastes pungent, bitter, and warm in nature; it enters the spleen and kidney meridians. Cnidium monnieri is the fruit of Cnidium monnieri (L.) Cuss., a plant of the genus Cnidium in the family Apiaceae. It has the effects of warming the kidney and strengthening yang, drying dampness and killing insects, and dispelling wind and relieving itching. It is mainly used to treat genital itching, eczema, scabies; cold and damp leucorrhea, dampness and low back pain; impotence due to kidney deficiency, infertility due to uterine cold, etc. The fruit of Cnidium monnieri contains 1.3% volatile oil, and 27 components have been separated from the oil. It also contains coumarins and other ingredients, such as cnidium monnieri and pepper toxin. Cnidium monnieri has an inhibitory effect on drug-resistant Staphylococcus aureus, Pseudomonas aeruginosa, dermatophytes, and Trichomonas.

[0049] Phellodendron chinense Schneid., also known as berberine, berberine bark, and phellodendron chinense, is bitter and cold in nature. It is good for the kidney and bladder. Phellodendron chinense Schneid. is a plant of the genus Phellodendron in the Rutaceae family. It has the effects of clearing away heat and dampness, purging fire and detoxifying, and removing bone steaming. It is mainly used for damp-heat diarrhea, hot stranguria, damp-heat diarrhea, jaundice, damp-heat beriberi, impotence, bone steaming and fever, night sweats, spermatorrhea, sores, swelling and poison, and eczema and itching. Phellodendron bark contains berberine, phellodendronine, magnolia alkaloids, jatrorrhizine, palmate leaf tetrandrine and other alkaloids, and contains phellodendron lactone, phellodendronone, phellodendron acid and 7-dehydrostigmasterol, β-sitosterol, campesterol, etc.; the bark of the phellodendron tree contains berberine, magnolia alkaloids, phellodendronine, palmate leaf tetrandrine and other alkaloids and lactones, sterols, etc. It has similar anti-pathogenic microbial effects as Coptis chinensis, and has inhibitory effects on various pathogenic bacteria such as Shigella dysenteriae, Salmonella typhi, Mycobacterium tuberculosis, Staphylococcus aureus, and hemolytic Streptococcus; it also has inhibitory effects on certain skin fungi, Leptospira, and hepatitis B surface antigen; the jatrorrhizine it contains has positive inotropic and antiarrhythmic effects similar to berberine; Phellodendron chinense extract has antihypertensive, anti-ulcer, sedative, muscle relaxant, blood sugar lowering and mouse antibody production promoting effects.

[0050] The present invention prepares a plant composition with vaginal antibacterial and anti-inflammatory effects by rationally combining the Chinese herbal medicines of Gardenia jasminoides, Saussurea involucrata, Agrimoniae herba, Stemona radix, Cnidium monnieri and Phellodendron chinense. Meanwhile, the present invention combines the microbial fermentation technology with the six kinds of Chinese herbal medicine plant compositions, which not only plays a synergistic role in the vaginal antibacterial and anti-inflammatory effects, but also brings the following advantages:

[0051] (1) The microbial fermentation preparation process adopted in the present invention is a biotransformation carried out under relatively mild conditions such as room temperature and pressure, which can protect the active ingredients in Chinese herbal medicines from destruction to the greatest extent possible, especially heat-sensitive aromatic volatile oils, vitamins and other active ingredients, and can more effectively protect them. It can also solve the problem that the active ingredients of traditional Chinese herbal medicines are difficult to extract to the greatest extent possible in traditional processes such as decocting, boiling, simmering, refining, steaming and soaking.

[0052] (2) The microbial fermentation preparation process adopted in the present invention allows the effective active ingredients in Chinese herbal medicines that cannot be directly utilized after entering the human body to be degraded into small molecular active substances through microbial fermentation, thereby promoting the absorption of the active ingredients of Chinese herbal medicines in the human body, greatly improving the efficacy of Chinese medicines, and improving the therapeutic effect; during the fermentation process, the Chinese herbal medicines change the content of their active ingredients under the action of microorganisms, and make the composition of the fermentation product different from that of the raw material medicine, and break the cell wall of the Chinese herbal medicine to dissolve the effective substances, thereby increasing the concentration of the active ingredients, and also improving the water solubility of the active ingredients of the Chinese herbal medicines, further promoting the absorption of the active ingredients of the Chinese herbal medicines in the human body.

[0053] (3) According to the microbial fermentation preparation process adopted by the present invention, some Chinese herbal medicines may produce new active substances, new functions and efficacy after being fermented by probiotics.

[0054] (4) The microbial fermentation preparation process adopted by the present invention can promote the growth of some Chinese herbal medicines, further increase the content of active ingredients, and play a synergistic role in the efficacy of the medicine.

[0055] (5) In the microbial fermentation preparation process adopted by the present invention, the biotransformation and decomposition action of the microorganisms can also decompose the toxic substances in the Chinese herbal medicines, or perform structural modification on the toxic active ingredients of the Chinese herbal medicines, thereby reducing the toxic side effects of the Chinese herbal medicines.

[0056] (6) The microbial fermentation preparation process adopted by the present invention has little irritation to the vagina, and long-term use will not destroy the balance of the vaginal flora, because the pharmaceutical composition uses probiotics Bacillus subtilis and Lactobacillus crispatus for fermentation.

[0057] The present invention is further described below in conjunction with examples, the purpose of which is to better understand the content of the present invention and reflect the essential characteristics of the present invention, so the examples cited should not be regarded as limiting the scope of protection of the present invention. It is also particularly pointed out here that the specific experimental methods and equipment involved in the examples are conventional methods or implemented under the conditions recommended by the manufacturer's instructions unless otherwise specified, and the reagents involved are commercially available unless otherwise specified.

[0058] Example 1

[0059] The Chinese medicinal materials Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri, and Phellodendron chinense were mixed in a ratio of 2:2:3:1:1:1 and crushed. After passing through an 80-mesh sieve, 100 g of the mixed powder was weighed and added to a triangular flask. Five times the amount of water was added and sterilized at 121 °C for 20 min. After cooling to room temperature, 15 ml of activated Bacillus subtilis (2.0 × 10 8 CFU·mL -1 ) and 5 ml of Lactobacillus crispatus seed solution (2.0×10 8 CFU·mL -1 ); ferment in a constant temperature incubator at 28°C for 5 days. After the fermentation, dry the fermented medicinal materials in a constant temperature oven at 60°C to constant weight, add 15 times of 50% ethanol to extract at 75°C for 1 hour, filter the residue by adding 10 times of 50% ethanol to extract at 75°C for 1 hour, filter the residue by combining the two filtrates, and filter using a 0.22μm filter membrane. After the filtrate is concentrated under reduced pressure, freeze-dried to obtain a dry fermented plant extract.

[0060] Comparative Example 1

[0061] The Chinese medicinal materials of Gardenia, Saussurea involucrata, Herba Agrimoniae, Radix Stemonae, Fructus Cnidii and Cortex Phellodendri were mixed in a ratio of 2:2:3:1:1:1, crushed and passed through an 80-mesh sieve. 100 g of the crushed Chinese herbal medicine was added with 15 times of 50% ethanol to extract at 75°C for 1 hour, filtered after the extraction, and the residue was further added with 10 times of 50% ethanol to extract at 75°C for 1 hour, filtered after the extraction, and the two filtrates were combined and filtered using a 0.22 μm filter membrane. After the filtrate was concentrated under reduced pressure, it was freeze-dried to obtain a dry plant extract.

[0062] Comparative Example 2

[0063] The Chinese medicinal materials Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri, and Phellodendron chinense were mixed in a ratio of 2:2:3:1:1:1 and crushed. After passing through an 80-mesh sieve, 100 g of the mixed powder was weighed and added to a triangular flask. Five times the amount of water was added and sterilized at 121 °C for 20 min. After cooling to room temperature, 15 ml of activated Bacillus subtilis (2.0 × 10 8 CFU·mL -1 ) and 5 ml of Lactobacillus crispatus seed solution (2.0×10 8 CFU·mL -1 ); after being fully stirred, the mixture was fermented in a constant temperature incubator at 28°C for 5 days. After the fermentation, the mixture was filtered using a 0.22 μm filter membrane. The filtrate was concentrated under reduced pressure and freeze-dried to obtain a dry fermented plant extract.

[0064] Comparative Example 3

[0065] The Chinese medicinal materials Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri, and Phellodendron chinense were mixed in a ratio of 2:2:3:1:1:1 and then crushed. The mixture was passed through an 80-mesh sieve. 100 g of the mixed powder was weighed and added with 15 times of 50% ethanol to extract at 75°C for 1 h. After the extraction, the mixture was filtered. The residue was further added with 10 times of 50% ethanol to extract at 75°C for 1 h. After the extraction, the mixture was filtered. The two filtrates were combined and concentrated under reduced pressure until there was no ethanol. The extract was sterilized at 121°C for 20 min. After cooling, 15 ml of activated Bacillus subtilis (2.0 × 10 8 CFU·mL -1 ) and 5 ml of Lactobacillus crispatus seed solution (2.0×10 8 CFU·mL -1 ); fermented in a constant temperature incubator at 28° C. for 5 days. After the fermentation, the fermented medicinal materials were concentrated under reduced pressure and freeze-dried to obtain a dry fermented plant extract.

[0066] Comparative Example 4

[0067] The Chinese medicinal materials Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri, and Phellodendron chinense were mixed in a ratio of 2:2:3:1:1:1 and crushed. After passing through an 80-mesh sieve, 100 g of the mixed powder was weighed and added to a triangular flask. Five times the amount of water was added and sterilized at 121 °C for 20 min. After cooling to room temperature, 15 ml of activated Bacillus subtilis (2.0 × 10 8 CFU·mL -1 ); ferment in a constant temperature incubator at 28°C for 5 days. After the fermentation is completed, the fermented medicinal materials are dried in a constant temperature oven at 60°C to constant weight, 15 times of 50% ethanol is added to extract at 75°C for 1 hour. After the extraction is completed, it is filtered, and the filter residue is further added with 10 times of 50% ethanol to extract at 75°C for 1 hour. After the extraction is completed, it is filtered, the two filtrates are combined, concentrated under reduced pressure, and freeze-dried to obtain a dry fermented plant extract.

[0068] Comparative Example 5

[0069] The Chinese medicinal materials Gardenia, Saussurea involucrata, Agrimoniae, Stemona, Cnidium monnieri, and Phellodendron chinense were mixed in a ratio of 2:2:3:1:1:1 and crushed, passed through an 80-mesh sieve, weighed 100 g of the mixed powder and added it to a triangular flask, added 5 times the amount of water, sterilized at 121 °C for 20 min, cooled to room temperature, and inoculated with 5 ml of activated Lactobacillus crispatus seed solution (2.0 × 10 8 CFU·mL -1 ); after being fully stirred, fermented in a constant temperature incubator at 28°C for 5 days. After the fermentation, the fermented medicinal materials were dried in a constant temperature oven at 60°C to constant weight, 15 times of 50% ethanol was added to extract at 75°C for 1 hour. After the extraction, it was filtered, and the filter residue was further added with 10 times of 50% ethanol to extract at 75°C for 1 hour. After the extraction, it was filtered, the two filtrates were combined, concentrated under reduced pressure, and freeze-dried to obtain a dry fermented plant extract.

[0070] Efficacy test:

[0071] 1. Antibacterial effect test:

[0072] Test bacteria: Staphylococcus aureus (ATCC 6538), Escherichia coli (8099), Candida albicans (ATCC10231).

[0073] Preparation of bacterial suspension: Take a 24-hour fresh slant culture of the bacteria, wash it with 5 ml of sterile PBS, and dilute it to a bacterial content of 10 4 -10 5 cfu / ml bacterial suspension for later use.

[0074] Antibacterial experiment: According to the microbroth dilution method, the test solution is sterilized by filtering with a microporous filter membrane, and diluted with the sterile liquid culture medium of the test strain according to the 2-fold dilution method to a dilution solution of 80-0.01 mg / mL. Take a microdilution plate (96 wells), add 50 μL of different concentrations of diluted sample solution and 50 μL of bacterial suspension to each well, and use 100 μL of culture medium containing bacteria without sample solution as a control, and make 3 parallels for each group. Escherichia coli and Staphylococcus aureus are cultured at 37°C for 18-24h, and Candida albicans is cultured at 25°C for 48h. Result determination: Observe the growth of bacteria in each concentration well of the 96-well plate. When there is bacterial growth (turbidity) in the control well, the growth of bacteria is completely inhibited compared with the control well. The lowest concentration corresponding to the clear culture medium is the minimum inhibitory concentration (MIC) of the composition.

[0075] The test results are shown in Table 1.

[0076] Table 1

[0077]

[0078] 2. Anti-inflammatory test:

[0079] To test whether the sample has the activity of inhibiting the release of inflammatory factors in the lipopolysaccharide-induced macrophage model, mouse mononuclear macrophage RAW264.7 was cultured in DMEM medium containing 10% inactivated fetal bovine serum, 100U / mL penicillin and 100μg / ml streptomycin, 5% CO2, 37℃. 5 / well were inoculated in a 96-well plate. After overnight in serum-free medium, the extracts of each group and the blank group were inoculated at a concentration of 100 μg / ml and treated for 2 h. Then, they were incubated with LPS at a concentration of 1 μg / ml for 24 h, and the cell supernatants were collected. The concentrations of various inflammatory factors (IL-6, TNF-α, NO) in the supernatants were detected by different kits.

[0080] The anti-inflammatory test results of the control group, Example 1, and Comparative Examples 1-5 are shown in Table 2.

[0081] Table 2

[0082] Group IL-6 (ng / L) TNFα(ng / L) NO(μM) Blank Group 0.13±0.01 3.08±0.17 0.57±0.04 Model Group 26.70±0.03 62.62±0.23 84.53±0.73 Experimental Example 1 12.56±0.07 40.25±0.21 59.24±0.86 Comparative Example 1 20.34±0.23 54.23±0.84 69.45±0.61 Comparative Example 2 18.32±0.45 52.72±0.16 65.29±1.06 Comparative Example 3 17.10±0.05 52.92±0.80 63.24±0.71 Comparative Example 4 15.56±0.07 49.57±1.19 61.56±0.92 Comparative Example 5 17.24±0.91 44.85±0.93 63.51±0.26

[0083] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects, characterized in that: It includes a fermentation product formed by fermenting a Chinese herbal medicine preparation and a compound probiotic. The raw materials of the Chinese herbal medicine preparation include gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron.

2. The Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects according to claim 1, characterized in that: The Chinese herbal medicine preparation is the powder of each raw material medicine, or the water or organic solvent extract of each raw material medicine, preferably the powder of each raw material medicine; the composite probiotics include a first probiotic and a second probiotic, the first probiotic is selected from one or more of lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), yeast fungi (Saccharomyces cerevisiae, Pichia pastoris), preferably Bacillus subtilis; the second probiotic is selected from one or more of Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii and Lactobacillus gasseri, preferably Lactobacillus crispatus.

3. The Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects according to claim 2, characterized in that: Among the raw materials of the Chinese herbal medicine preparation, the mass ratio of Gardenia jasminoides, Saussurea involucrata, Herba Agrimoniae, Radix Stemonae, Fructus Cnidii and Phellodendron amurense is (1-3): (1-3): (1-5): 1:1:1, and the particle size of the powder of each raw material of the Chinese herbal medicine preparation is 40-200 mesh; the bacterial count ratio of the first probiotic and the second probiotic is (1-5):

1.

4. A method for preparing a traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects as claimed in any one of claims 1 to 3, characterized in that: The steps include: Step (1), weighing gardenia, snow lotus, agrimony, stemona, cnidium monnieri and phellodendron amurense according to the proportion, crushing, and passing through an 80-mesh sieve to obtain a mixed powder of Chinese herbal medicines; Step (2), weighing a certain amount of mixed powder of Chinese herbal medicine, adding 5-10 times of water, stirring evenly and sterilizing at 121° C.; Step (3), after cooling, inoculating the composite probiotic strain in an ultra-clean workbench, and fermenting at 25°C-30°C for 3d-7d; Step (4), sterilizing after the fermentation is completed, drying the fermented medicinal materials in a 60°C constant temperature oven to constant weight, adding 15 times of 50% ethanol to extract at 75°C for 1h, filtering after the extraction is completed, adding 10 times of 50% ethanol to the filter residue to extract at 75°C for 1h, filtering after the extraction is completed, combining the two filtrates, filtering with a 0.22μm filter membrane, and decompressing and concentrating the filtrate for freeze-drying to obtain a dry fermented plant extract.

5. The preparation method according to claim 5, characterized in that: The composite probiotics include a first probiotic and a second probiotic, wherein the first probiotic is selected from one or more of lactic acid bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus), Bacillus (Bacillus licheniformis, Bacillus subtilis, Bacillus cereus), and yeast fungi (Saccharomyces cerevisiae, Pichia pastoris), preferably Bacillus subtilis; and the second probiotic is selected from one or more of Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii, and Lactobacillus gasseri, preferably Lactobacillus crispatus.

6. The preparation method according to claim 5, characterized in that: For every 100 g of the mixed powder of Chinese herbal medicine, the volume of the inoculated composite probiotic seed solution is 10-30 mL.

7. Use of a traditional Chinese medicine composition having vaginal antibacterial and anti-inflammatory effects as claimed in any one of claims 1 to 3 and a traditional Chinese medicine composition prepared by the preparation method as claimed in any one of claims 4 to 6 in preparing products for preventing or treating vaginal antibacterial and anti-inflammatory effects.

8. An external preparation having vaginal antibacterial and anti-inflammatory effects, characterized in that: The invention comprises the traditional Chinese medicine composition with vaginal antibacterial and anti-inflammatory effects as claimed in any one of claims 1 to 3 and the traditional Chinese medicine composition prepared by the preparation method as claimed in any one of claims 4 to 6.

9. The external preparation with vaginal antibacterial and anti-inflammatory effects according to claim 8, characterized in that: The dosage form of the external preparation with vaginal antibacterial and anti-inflammatory effects includes ointment, spray, douche or suppository.

10. The external preparation with vaginal antibacterial and anti-inflammatory effects according to claim 8, characterized in that: The external preparation with vaginal antibacterial and anti-inflammatory effects also includes at least one of an excipient, a bactericide, a moisturizer, an anti-allergic agent, a chelating agent and a flavor.

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