A pharmaceutical composition for treating gynecological inflammation and a preparation method thereof

Abstract

The application provides a pharmaceutical composition for treating gynecological inflammation and a preparation method thereof, and belongs to the technical field of medicines. After being subjected to water extraction and alcohol extraction, Sophora flavescens, Phellodendri chinensis, Forsythia suspensa and Ligustrum lucidum are used to obtain traditional Chinese medicine water extracts and traditional Chinese medicine alcohol extracts; the traditional Chinese medicine water extracts are reacted with silver salt and copper salt to obtain traditional Chinese medicine water extract-metal ion complex; the traditional Chinese medicine water extract-metal ion complex is mixed with the traditional Chinese medicine alcohol extracts, sodium taurodeoxycholate, laurocapram and berberine to prepare nanometer drug delivery micelles; and the nanometer drug delivery micelles are added into a hydrogel of chitosan, poloxamer P407 and poloxamer P188 to prepare the pharmaceutical composition for treating gynecological inflammation. The prepared composition has excellent treatment effect and low toxicity, thereby reducing the cost and improving the treatment effect of gynecological diseases such as chronic pelvic inflammatory disease, chronic cervicitis, vaginitis and artificial abortion complications, and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a pharmaceutical composition for treating gynecological inflammation and its preparation method. Background Technology

[0002] Gynecological diseases, which trouble many people, have seen a significant increase in incidence due to factors such as environmental pollution, competitive pressure, fast-paced work, and changes in lifestyle, and are showing a trend towards affecting younger people. Because gynecological diseases are common and frequently occurring, a large number of people require medication, and the use of gynecological medications in my country is increasing year by year.

[0003] There are many types of drugs for treating gynecological inflammation. For example, CN101284066A discloses a suppository for treating gynecological inflammation, which is prepared by conventional preparation methods using traditional Chinese medicine raw materials in the following weight ratios: 20-100 parts of Phellodendron bark, 30-100 parts of Sophora flavescens root, 20-90 parts of Kochia scoparia fruit, 10-40 parts of Agrimonia pilosa root, and 0.4-3 parts of borneol. CN101185702A discloses a drug for treating gynecological inflammation, which is a drug made from traditional Chinese medicine raw materials, including Carthamus tinctorius, Ligusticum chuanxiong rhizome, Taraxacum mongolicum, Viola yedoensis root, Scrophularia ningpoensis root, Leonurus japonicus root, Cnidium monnieri fruit, Lycium barbarum fruit, and Cordyceps sinensis. CN101773630A and CN1857428A also disclose drug compositions for treating gynecological inflammation.

[0004] CN102178704A discloses a drug with dual anti-inflammatory and anti-cancer effects, made from yew essential oil and yew extract obtained by physical extraction. This yew essential oil and yew extract can be directly applied to pharmaceutical manufacturing processes for oral or topical anti-inflammatory and anti-cancer preparations, such as capsules, emulsions, suspensions, powders, and foams.

[0005] However, the above-mentioned compositions have problems such as poor efficacy, slow onset of action, short effective drug residence time, and poor stability.

[0006] Currently, Western medicine generally uses antibiotics to treat this disease. However, because antibiotics have difficulty penetrating the affected area to exert their effects, they are less effective at eliminating inflammation in fibrous and connective tissues. Furthermore, antibiotics lack the ability to relieve adhesions and alleviate pain. Therefore, using antibiotics alone to treat chronic pelvic inflammatory disease is not always satisfactory, and it often leads to recurrence, increased drug resistance, and difficulty in achieving a complete cure. Traditional Chinese medicine (TCM) has significant advantages and unique characteristics in treating this disease. Currently, TCM treatment mainly includes internal and external therapies. Internal therapies utilize syndrome differentiation and treatment based on menstrual cycle regulation to achieve the effects of clearing heat and detoxifying, nourishing qi and blood, soothing the liver and relieving depression, removing blood stasis and regulating menstruation, and reducing inflammation and relieving pain. These methods have shown good results in treating chronic gynecological inflammation. Summary of the Invention

[0007] The purpose of this invention is to provide a pharmaceutical composition for treating gynecological inflammation and its preparation method, which has excellent therapeutic effects, low toxicity, thereby reducing costs and improving the treatment effect of gynecological diseases such as chronic pelvic inflammatory disease, chronic cervicitis, vaginitis, and complications of induced abortion, and has broad application prospects.

[0008] The technical solution of this invention is implemented as follows:

[0009] This invention provides a method for preparing a pharmaceutical composition for treating gynecological inflammation. The method involves extracting Sophora flavescens, Phellodendron chinense, Forsythia suspensa, and Ligustrum lucidum with water and alcohol to obtain aqueous and alcoholic extracts of the traditional Chinese medicine. The aqueous extracts are then reacted with silver and copper salts to obtain a water-metal ion complex. This complex is then mixed with the alcoholic extract, sodium taurine deoxycholate, laurocapram, and berberine to prepare nano-drug delivery micelles. These micelles are then added to a hydrogel containing chitosan, poloxamer P407, and poloxamer P188 to obtain the pharmaceutical composition for treating gynecological inflammation.

[0010] As a further improvement to the present invention, the following steps are included:

[0011] S1. Water extraction of traditional Chinese medicine: Sophora flavescens, Phellodendron chinense, Forsythia suspensa and Ligustrum lucidum were washed, dried and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, heated to boiling and extracted. The powder was filtered, the solid was kept for use, and the filtrate was dried to obtain water extract of traditional Chinese medicine.

[0012] S2. Alcohol extraction of traditional Chinese medicine: The solid is added to an ethanol solution, heated for extraction, filtered, and the filtrate is dried to obtain an alcohol extract of traditional Chinese medicine;

[0013] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve the water extract of traditional Chinese medicine obtained in step S1 in water, add silver salt and copper salt, heat and stir to react, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0014] S4. Preparation of nano-drug delivery micelles: The alcoholic extract of traditional Chinese medicine obtained in step S2, the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, sodium taurine deoxycholate, laurocapram, and berberine were added to an ethanol solution, stirred and mixed evenly, the solvent was removed by vacuum evaporation, water was added and shaken, the supernatant was removed by centrifugation, mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0015] S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Chitosan is dissolved in acid, poloxamer P407 and poloxamer P188 are added and mixed to expand, and the nano-drug delivery micelles obtained in step S4 are added and stirred to mix evenly to obtain a pharmaceutical composition for treating gynecological inflammation.

[0016] As a further improvement of the present invention, the mass ratio of Sophora flavescens, Phellodendron chinense, Forsythia suspensa and Ligustrum lucidum in step S1 is 5-7:2-4:1-3:3-5, the solid-liquid ratio of the Chinese herbal powder and water is 1:5-7 g / mL, and the extraction time is 3-5 h.

[0017] As a further improvement of the present invention, the solid-liquid ratio of the solid and ethanol solution in step S2 is 1:3-5 g / mL, the concentration of the ethanol solution is 50-60 wt%, and the heating extraction time is 2-4 h.

[0018] As a further improvement of the present invention, the mass ratio of the herbal water extract, silver salt and copper salt in step S3 is 100:3-5:1-2, the silver salt is silver nitrate, the copper salt is at least one of copper chloride, copper sulfate and copper nitrate, and the heating and stirring reaction temperature is 35-45℃ and the time is 0.5-1h.

[0019] As a further improvement of the present invention, the mass ratio of the Chinese herbal alcohol extract, the Chinese herbal water extract-metal ion complex, sodium taurine deoxycholate, laurocapram, berberine, and mannitol aqueous solution in step S4 is 10-15:12-17:150-170:15-20:3-5:15-20, the concentration of the mannitol aqueous solution is 2-3 wt%, and the shaking time is 1-3 h.

[0020] As a further improvement of the present invention, the concentration of the acid solution in step S5 is 2-4 wt%, the mass ratio of chitosan, nano-drug micelles, poloxamer P407 and poloxamer P188 is 3-5:2-3:40-50:20-25, the expansion temperature is 3-5℃, and the time is 20-24h.

[0021] As a further improvement to the present invention, the specific steps include:

[0022] S1. Water extraction of traditional Chinese medicine: Wash, dry and pulverize 5-7 parts by weight of Sophora flavescens, 2-4 parts by weight of Phellodendron chinense, 1-3 parts by weight of Forsythia suspensa and 3-5 parts by weight of Ligustrum lucidum to obtain traditional Chinese medicine powder. Add water to the powder, with a solid-liquid ratio of 1:5-7 g / mL. Heat to boiling and extract for 3-5 hours. Filter, keep the solid, and dry the filtrate to obtain water extract of traditional Chinese medicine.

[0023] S2. Alcohol extraction of traditional Chinese medicine: The solid is added to a 50-60 wt% ethanol solution, the solid-liquid ratio of the solid to the 50-60 wt% ethanol solution is 1:3-5 g / mL, the mixture is heated and extracted for 2-4 hours, filtered, and the filtrate is dried to obtain an alcohol extract of traditional Chinese medicine.

[0024] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 3-5 parts by weight of silver salt and 1-2 parts by weight of copper salt, heat to 35-45℃, stir and react for 0.5-1h, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0025] S4. Preparation of nano-drug delivery micelles: 10-15 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 12-17 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 150-170 parts by weight of sodium taurine deoxycholate, 15-20 parts by weight of lauryl azone, and 3-5 parts by weight of berberine were added to 500 parts by weight of 40-50 wt% ethanol solution, stirred and mixed evenly, the solvent was removed by vacuum evaporation, water was added and shaken for 1-3 h, the supernatant was removed by centrifugation, 15-20 parts by weight of 2-3 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0026] S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 3-5 parts by weight of chitosan in 200 parts by weight of 2-4 wt% acetic acid solution, add 40-50 parts by weight of poloxamer P407 and 20-25 parts by weight of poloxamer P188 and mix, expand at 3-5℃ for 20-24 h, add 2-3 parts by weight of the nano-drug delivery micelles obtained in step S4 and stir to mix evenly to obtain a pharmaceutical composition for treating gynecological inflammation.

[0027] The present invention provides a pharmaceutical composition for treating gynecological inflammation prepared by the above-described preparation method.

[0028] This invention provides the application of the above-mentioned pharmaceutical composition for treating gynecological inflammation in the treatment of vaginitis, pelvic inflammatory disease, cervicitis, adnexitis, and vulvitis.

[0029] The present invention has the following beneficial effects:

[0030] The herbal components of this invention include Sophora flavescens, Phellodendron chinense, Forsythia suspensa, and Ligustrum lucidum. Sophora flavescens is rich in active ingredients including polysaccharides, flavonoids (matrine, isochoride, sophorone G, etc.), and alkaloids (oxymatrine, sophoridine, etc.), which can exert various pharmacological effects such as inhibiting proliferation, antibacterial, anti-inflammatory, immunomodulatory, antiviral, anti-apoptotic, and antitumor effects through pathways such as nuclear factor κB and the phosphatidylinositol 3-kinase / protein kinase B / target of rapamycin signaling pathway. Phellodendron chinense contains abundant flavonoids, polyphenols, and alkaloids, which inhibit inflammation through the NF-κB and mitogen-activated protein kinase signaling pathways and downregulate nitric oxide and inducible nitric oxide synthase, thereby further reducing the occurrence of inflammatory responses. Forsythia suspensa contains forsythoside and volatile oil, which have strong inhibitory effects and antibacterial and antipyretic effects against Staphylococcus aureus. Rhodioloside, tyrosol, oleanolic acid, and ursolic acid in privet fruit have anti-inflammatory, antibacterial, white blood cell-increasing, non-specific immune-enhancing, and immunomodulatory effects, and possess anti-inflammatory and antimicrobial pharmacological functions.

[0031] The aqueous extract obtained from the herbal components of this invention contains abundant polysaccharides, proteins, and other active components, exhibiting good antibacterial and anti-inflammatory effects. Simultaneously, it contains abundant hydroxyl, amino, and carboxyl groups, which can form complexes with metal ions (silver and copper ions), thus forming a temperature-sensitive complex of the herbal aqueous extract and the metal ion complex; Ag + It can also effectively kill pathogenic microorganisms, Cu 2+ It has strong oxidizing properties and will cause bacteria to die when it comes into contact with them. It can oxidize the biochemical substances such as proteins, nucleic acids and enzymes in bacteria, causing cell death. The synergy between the two can also effectively prevent pathogenic microorganisms from developing drug resistance.

[0032] Chitosan possesses good biodegradability, biocompatibility, and bacterial resistance; however, its poor water solubility hinders the formation of hydrogel network structures. Chitosan can crosslink with metal ions (silver and copper ions), resulting in rapid gelation, a simple gelation process, long-lasting shape retention, and significant plasticity, and it can undergo a gel-sol transition. The resulting thermosensitive hydrogel can respond to changes in external temperature, accelerating drug release and delivery, while also exhibiting biodegradability, biocompatibility, and bacterial resistance.

[0033] The sodium taurocholate nanomicelles prepared in this invention are biosurfactants that can improve the solubility of insoluble drugs such as berberine and poorly soluble components in traditional Chinese medicine extracts. They also act as penetration enhancers, increasing drug absorption through mucous membranes. Furthermore, the nanomicelle drug-carrying system formed by sodium taurocholate and laurocapram may better facilitate drug penetration through mucosal layers and cell membranes, while also forming stable nanomicelles, facilitating subsequent hydrogel preparation. This invention adds berberine to the traditional Chinese medicine components, resulting in a synergistic effect that significantly improves the therapeutic efficacy of the composition while reducing side effects and minimizing drug resistance in pathogenic bacteria.

[0034] Due to the vagina's self-cleaning function, traditional vaginal medications suffer from limitations such as poor drug distribution, poor adhesion, and short retention time. The pharmaceutical composition for treating gynecological inflammation prepared in this invention, while possessing antibacterial, anti-inflammatory, heat-clearing, and detoxifying properties, also promotes blood circulation, removes blood stasis, relieves pain, improves tissue microcirculation, and facilitates the dissipation of inflammation. Furthermore, the prepared pharmaceutical composition for treating gynecological inflammation is a thermosensitive hydrogel. When it reaches its gelation temperature, it transforms into a gel. Its low viscosity at room temperature facilitates vaginal administration and optimal diffusion within the mucosa, prolonging the drug's retention time in the vagina. As it gradually warms to its gelation temperature (forming a gel at body temperature), it forms a gel state, further extending the administration time. The sodium taurodeoxycholate and lauryl azone in the micelle particles promote transdermal absorption, enhancing efficacy and prolonging the administration time. This results in excellent therapeutic effects even at relatively low drug concentrations, low toxicity, reduced costs, and improved treatment outcomes for gynecological diseases such as chronic pelvic inflammatory disease, chronic cervicitis, vaginitis, and complications from induced abortion, demonstrating broad application prospects. Detailed Implementation

[0035] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Chitosan, purity >98%, purchased from Shandong Jianyou Bioengineering Co., Ltd.

[0037] Example 1

[0038] This embodiment provides a method for preparing a pharmaceutical composition for treating gynecological inflammation, specifically including the following steps:

[0039] S1. Water extraction of traditional Chinese medicine: 5 parts by weight of Sophora flavescens, 2 parts by weight of Phellodendron chinense, 1 part by weight of Forsythia suspensa, and 3 parts by weight of Ligustrum lucidum were washed, dried, and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, and the solid-liquid ratio of the powder to water was 1:5 g / mL. The mixture was heated to boiling and extracted for 3 hours. The mixture was filtered, the solid was retained, and the filtrate was dried to obtain a water extract of traditional Chinese medicine.

[0040] S2. Alcohol extraction of traditional Chinese medicine: The solid is added to a 50wt% ethanol solution, the solid-liquid ratio of the solid to the 50wt% ethanol solution is 1:3 g / mL, heated to 50℃, extracted for 2 hours, filtered, and the filtrate is dried to obtain an alcohol extract of traditional Chinese medicine.

[0041] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 3 parts by weight of silver nitrate and 1 part by weight of copper chloride, heat to 35°C, stir and react for 0.5 h, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0042] S4. Preparation of nano-drug delivery micelles: 10 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 12 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 150 parts by weight of sodium taurine deoxycholate, 15 parts by weight of lauryl azone, and 3 parts by weight of berberine were added to 500 parts by weight of 40 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 1 h, the supernatant was removed by centrifugation, 15 parts by weight of 2 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0043] S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 3 parts by weight of chitosan in 200 parts by weight of 2wt% acetic acid solution, add 40 parts by weight of poloxamer P407 and 20 parts by weight of poloxamer P188 and stir for 10 min, expand at 3°C ​​for 20 h, add 2 parts by weight of the nano-drug delivery micelles prepared in step S4 and stir for 20 min to obtain a pharmaceutical composition for treating gynecological inflammation.

[0044] Example 2

[0045] This embodiment provides a method for preparing a pharmaceutical composition for treating gynecological inflammation, specifically including the following steps:

[0046] S1. Water extraction of traditional Chinese medicine: 7 parts by weight of Sophora flavescens, 4 parts by weight of Phellodendron chinense, 3 parts by weight of Forsythia suspensa and 5 parts by weight of Ligustrum lucidum were washed, dried and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, and the solid-liquid ratio of the powder to water was 1:7 g / mL. The mixture was heated to boiling and extracted for 5 hours. The mixture was filtered, the solid was retained, and the filtrate was dried to obtain water extract of traditional Chinese medicine.

[0047] S2. Alcohol extraction of traditional Chinese medicine: The solid is added to a 60wt% ethanol solution, the solid-liquid ratio of the solid to the 60wt% ethanol solution is 1:5 g / mL, heated to 50℃, extracted for 4 h, filtered, and the filtrate is dried to obtain an alcohol extract of traditional Chinese medicine.

[0048] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 5 parts by weight of silver nitrate and 2 parts by weight of copper sulfate, heat to 45°C, stir and react for 1 hour, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0049] S4. Preparation of nano-drug delivery micelles: 15 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 17 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 170 parts by weight of sodium taurine deoxycholate, 20 parts by weight of lauryl azone, and 5 parts by weight of berberine were added to 500 parts by weight of 50 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 3 h, the supernatant was removed by centrifugation, 20 parts by weight of 3 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0050] S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 5 parts by weight of chitosan in 200 parts by weight of 4 wt% acetic acid solution, add 50 parts by weight of poloxamer P407 and 25 parts by weight of poloxamer P188 and stir for 10 min, expand at 5°C for 24 h, add 3 parts by weight of the nano-drug delivery micelles prepared in step S4 and stir for 20 min to obtain a pharmaceutical composition for treating gynecological inflammation.

[0051] Example 3

[0052] This embodiment provides a method for preparing a pharmaceutical composition for treating gynecological inflammation, specifically including the following steps:

[0053] S1. Water extraction of traditional Chinese medicine: 6 parts by weight of Sophora flavescens, 3 parts by weight of Phellodendron chinense, 2 parts by weight of Forsythia suspensa, and 4 parts by weight of Ligustrum lucidum were washed, dried, and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, and the solid-liquid ratio of the powder to water was 1:6 g / mL. The mixture was heated to boiling and extracted for 4 hours. The mixture was filtered, the solid was retained, and the filtrate was dried to obtain a water extract of traditional Chinese medicine.

[0054] S2. Alcohol extraction of traditional Chinese medicine: The solid was added to a 55wt% ethanol solution, the solid-liquid ratio of the solid to the 5wt% ethanol solution was 1:4 g / mL, heated to 50℃, extracted for 3 h, filtered, and the filtrate was dried to obtain an alcohol extract of traditional Chinese medicine.

[0055] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 4 parts by weight of silver nitrate and 1.5 parts by weight of copper nitrate, heat to 40°C, stir and react for 1 hour, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0056] S4. Preparation of nano-drug delivery micelles: 12 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 15 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 160 parts by weight of sodium taurine deoxycholate, 17 parts by weight of lauryl azone, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0057] S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 4 parts by weight of chitosan in 200 parts by weight of 3 wt% acetic acid solution, add 45 parts by weight of poloxamer P407 and 22 parts by weight of poloxamer P188 and stir for 10 min, expand at 4°C for 22 h, add 2.5 parts by weight of the nano-drug delivery micelles prepared in step S4 and stir for 20 min to obtain a pharmaceutical composition for treating gynecological inflammation.

[0058] Comparative Example 1

[0059] The difference between Example 3 and Example 4 is that silver nitrate was not added in step S3. Specifically:

[0060] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight, add 5.5 parts by weight of copper nitrate, heat to 40°C, stir and react for 1 hour, and dry to obtain water extract of traditional Chinese medicine-metal ion complex.

[0061] Comparative Example 2

[0062] The difference from Example 3 is that copper nitrate was not added in step S3. Specifically:

[0063] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight, add 5.5 parts by weight of silver nitrate, heat to 40°C, stir and react for 1 hour, and dry to obtain water extract of traditional Chinese medicine-metal ion complex.

[0064] Comparative Example 3

[0065] The difference from Example 3 is that step S3 is not performed. Specifically:

[0066] S1. Water extraction of traditional Chinese medicine: 6 parts by weight of Sophora flavescens, 3 parts by weight of Phellodendron chinense, 2 parts by weight of Forsythia suspensa, and 4 parts by weight of Ligustrum lucidum were washed, dried, and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, and the solid-liquid ratio of the powder to water was 1:6 g / mL. The mixture was heated to boiling and extracted for 4 hours. The mixture was filtered, the solid was retained, and the filtrate was dried to obtain a water extract of traditional Chinese medicine.

[0067] S2. Alcohol extraction of traditional Chinese medicine: The solid was added to a 55wt% ethanol solution, the solid-liquid ratio of the solid to the 5wt% ethanol solution was 1:4 g / mL, heated to 50℃, extracted for 3 h, filtered, and the filtrate was dried to obtain an alcohol extract of traditional Chinese medicine.

[0068] S3. Preparation of nano-drug delivery micelles: 27 parts by weight of the Chinese herbal alcohol extract obtained in step S2, 15 parts by weight of the Chinese herbal alcohol extract obtained in step S2, 160 parts by weight of sodium taurine deoxycholate, 17 parts by weight of lauryl azone, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0069] S4. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 4 parts by weight of chitosan in 200 parts by weight of 3wt% acetic acid solution, add 45 parts by weight of poloxamer P407 and 22 parts by weight of poloxamer P188 and stir for 10 min, expand at 4°C for 22 h, add 2.5 parts by weight of the nano-drug delivery micelles prepared in step S3 and stir for 20 min to obtain a pharmaceutical composition for treating gynecological inflammation.

[0070] Comparative Example 4

[0071] The difference from Example 3 is that lauryl azone was not added in step S4. Specifically:

[0072] S4. Preparation of nano-drug delivery micelles: 12 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 15 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 177 parts by weight of sodium taurine deoxycholate, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0073] Comparative Example 5

[0074] The difference from Example 3 is that berberine was not added in step S4. Specifically:

[0075] S4. Preparation of nano-drug delivery micelles: 12 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 15 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 160 parts by weight of sodium taurine deoxycholate, and 17 parts by weight of lauryl azone were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0076] Comparative Example 6

[0077] The difference compared to Example 3 is that no traditional Chinese medicine alcohol extract was added in step S4. Specifically:

[0078] S4. Preparation of nano-drug delivery micelles: 15 parts by weight of the water extract-metal ion complex of traditional Chinese medicine obtained in step S3, 160 parts by weight of sodium taurine deoxycholate, 17 parts by weight of lauryl azone, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0079] Comparative Example 7

[0080] The difference compared to Example 3 is that the herbal water extract-metal ion complex was not added in step S4. Specifically:

[0081] S4. Preparation of nano-drug delivery micelles: 12 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 160 parts by weight of sodium taurine deoxycholate, 17 parts by weight of lauryl azone, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles.

[0082] Comparative Example 8

[0083] The difference from Example 3 is that step S5 is not performed. Specifically:

[0084] S1. Water extraction of traditional Chinese medicine: 6 parts by weight of Sophora flavescens, 3 parts by weight of Phellodendron chinense, 2 parts by weight of Forsythia suspensa, and 4 parts by weight of Ligustrum lucidum were washed, dried, and pulverized to obtain traditional Chinese medicine powder. The powder was added to water, and the solid-liquid ratio of the powder to water was 1:6 g / mL. The mixture was heated to boiling and extracted for 4 hours. The mixture was filtered, the solid was retained, and the filtrate was dried to obtain a water extract of traditional Chinese medicine.

[0085] S2. Alcohol extraction of traditional Chinese medicine: The solid was added to a 55wt% ethanol solution, the solid-liquid ratio of the solid to the 5wt% ethanol solution was 1:4 g / mL, heated to 50℃, extracted for 3 h, filtered, and the filtrate was dried to obtain an alcohol extract of traditional Chinese medicine.

[0086] S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 4 parts by weight of silver nitrate and 1.5 parts by weight of copper nitrate, heat to 40°C, stir and react for 1 hour, dry, and obtain water extract of traditional Chinese medicine-metal ion complex.

[0087] S4. Preparation of nano-drug delivery micelles: 12 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 15 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 160 parts by weight of sodium taurine deoxycholate, 17 parts by weight of lauryl azone, and 4 parts by weight of berberine were added to 500 parts by weight of 45 wt% ethanol solution, stirred and mixed for 20 min, the solvent was removed by vacuum evaporation, water was added and shaken for 2 h, the supernatant was removed by centrifugation, 17 parts by weight of 2.5 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles, which is the drug composition for treating gynecological inflammation.

[0088] Test Example 1

[0089] The pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 and Comparative Examples 1-8 were freeze-dried and cut into round slices with a diameter of 5 mm and a thickness of 1 mm. The original sample slices were placed on Luria-Bertani medium inoculated with bacterial colonies and incubated at 35°C for 24 h. The diameter of the inhibition zone was tested. The bacterial strains used were Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC29740), Aerobic Escherichia coli (ATCC27325), and Candida albicans (ATCC60193). The results are shown in Table 1.

[0090] Table 1

[0091]

[0092]

[0093] As shown in the table above, the pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 of this invention have good inhibitory effects on Escherichia coli, Staphylococcus aureus, Escherichia coli aerogenes, and Candida albicans.

[0094] Test Example 2

[0095] The sol-gel transition of the pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 and Comparative Example 8 at room temperature (25°C) and 37°C was detected using the inverted test tube method, and the gelation time was determined. The results are shown in Table 2.

[0096] Table 2

[0097]

[0098] As shown in the table above, the pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 of the present invention are in a solution state at room temperature and can quickly form a gel at a body temperature of 37°C, thereby improving the vaginal adhesion effect and prolonging the administration time.

[0099] Test Example 3

[0100] Mice were randomly divided into a blank control group, a model group, a positive model group, Example 1-3 groups, and Comparative Examples 1-8 groups, with 10 mice in each group. Except for the blank control group, bacterial vaginosis was modeled in the other groups by injecting 0.3 mL of a mixed bacterial solution of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa (1:1:1) into the vagina of mice once a day for 7 consecutive days. After that, obvious redness and swelling and purulent discharge were visible at the vaginal opening of the mice.

[0101] After successful membrane formation, the positive group, Examples 1-3, and Comparative Examples 1-8 were treated with local vaginal medication. The positive group received 0.2 mL of levofloxacin solution (0.5 mg / mL) topically, once daily for 7 consecutive days. Examples 1-3 and Comparative Examples 1-10 (each group received 2 g of the corresponding gynecological inflammation treatment drug composition) topically, once daily for 7 consecutive days. Medication was administered via vaginal injection to mice. The model group and control group received an equal volume of physiological saline vaginally (2 g once daily for 7 consecutive days).

[0102] 1. Vaginal condition in mice

[0103] After successful membrane formation and 7 days of group feeding, vaginal secretion samples were collected from mice for smear microscopic examination. The Nugent score was used to evaluate vaginal condition: 0 points indicated the best vaginal environment, 10 points the worst, and >4 points indicated the onset of vaginal inflammation, with the degree of inflammation increasing with the score. The results are shown in Table 3.

[0104] Table 3

[0105]

[0106] Note: * indicates P < 0.05 compared to the control group; # indicates P < 0.05 compared to the model group.

[0107] As can be seen from the table above, the Nugent score of the pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 of the present invention is significantly reduced.

[0108] 2. Blood collection and inflammatory factor detection

[0109] After scoring, orbital venous blood was collected from each group of mice, and the supernatant serum was obtained by centrifugation. The concentrations of IL-6 and CRP (C-reactive protein) in the mouse serum were detected using a kit. The results are shown in Table 4.

[0110] Table 4

[0111] Group IL-6 concentration (pg / mL) CRP concentration (pg / mL) Blank group 13.19±2.51 171.24±22.51 Model group 22.15±5.29* 655.15±49.26* Positive drug group 19.22±3.92# 95.16±17.26# Example 1 15.62±2.29# 80.11±14.15# Example 2 15.09±3.98# 81.09±16.21# Example 3 15.26±2.88# 79.18±15.11# Comparative Example 1 16.22±2.17 89.26±11.29 Comparative Example 2 16.72±2.58 86.77±13.18 Comparative Example 3 17.45±3.19 95.26±16.38 Comparative Example 4 17.89±1.85 97.18±13.58 Comparative Example 5 18.72±2.05 220.51±15.28 Comparative Example 6 18.59±2.25 115.26±14.22 Comparative Example 7 18.94±2.11 125.26±13.09 Comparative Example 8 19.03±1.95 133.17±12.18

[0112] Note: * indicates P < 0.05 compared to the control group; # indicates P < 0.05 compared to the model group.

[0113] Multicellular pro-inflammatory cytokine IL-6 promotes inflammation. CRP is a non-specific acute-phase reactive protein and an inflammatory marker. As shown in the table above, the concentrations of IL-6 and CRP in the pharmaceutical compositions for treating gynecological inflammation prepared in Examples 1-3 of this invention are significantly reduced, indicating a better anti-inflammatory effect.

[0114] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a pharmaceutical composition for treating gynecological inflammation, characterized in that, Specifically, the following steps are included: S1. Water extraction of traditional Chinese medicine: Wash, dry and pulverize 5-7 parts by weight of Sophora flavescens, 2-4 parts by weight of Phellodendron chinense, 1-3 parts by weight of Forsythia suspensa and 3-5 parts by weight of Ligustrum lucidum to obtain traditional Chinese medicine powder. Add water to the powder, with a solid-liquid ratio of 1:5-7 g / mL. Heat to boiling and extract for 3-5 hours. Filter, keep the solid, and dry the filtrate to obtain water extract of traditional Chinese medicine. S2. Alcohol extraction of traditional Chinese medicine: The solid is added to a 50-60 wt% ethanol solution, the solid-liquid ratio of the solid to the 50-60 wt% ethanol solution is 1:3-5 g / mL, the mixture is heated and extracted for 2-4 hours, filtered, and the filtrate is dried to obtain an alcohol extract of traditional Chinese medicine. S3. Preparation of water extract of traditional Chinese medicine-metal ion complex: Dissolve 100 parts by weight of water extract of traditional Chinese medicine obtained in step S1 in 500 parts by weight of water, add 3-5 parts by weight of silver salt and 1-2 parts by weight of copper salt, heat to 35-45℃, stir and react for 0.5-1h, dry, and obtain water extract of traditional Chinese medicine-metal ion complex. S4. Preparation of nano-drug delivery micelles: 10-15 parts by weight of the alcoholic extract of traditional Chinese medicine obtained in step S2, 12-17 parts by weight of the aqueous extract of traditional Chinese medicine-metal ion complex obtained in step S3, 150-170 parts by weight of sodium taurine deoxycholate, 15-20 parts by weight of lauryl azone, and 3-5 parts by weight of berberine were added to 500 parts by weight of 40-50 wt% ethanol solution, stirred and mixed evenly, the solvent was removed by vacuum evaporation, water was added and shaken for 1-3 hours, the supernatant was removed by centrifugation, 15-20 parts by weight of 2-3 wt% mannitol aqueous solution was added, and the mixture was freeze-dried to obtain nano-drug delivery micelles. S5. Preparation of a pharmaceutical composition for treating gynecological inflammation: Dissolve 3-5 parts by weight of chitosan in 200 parts by weight of 2-4 wt% acetic acid solution, add 40-50 parts by weight of poloxamer P407 and 20-25 parts by weight of poloxamer P188 and mix, expand at 3-5℃ for 20-24 h, add 2-3 parts by weight of the nano-drug delivery micelles obtained in step S4 and stir to mix evenly to obtain a pharmaceutical composition for treating gynecological inflammation.

2. The preparation method according to claim 1, characterized in that, The silver salt mentioned in step S3 is silver nitrate, and the copper salt is at least one of copper chloride, copper sulfate, and copper nitrate.

3. A pharmaceutical composition for treating gynecological inflammation prepared by the method described in claim 1 or 2.

4. The use of the pharmaceutical composition for treating gynecological inflammation as described in claim 3 in the preparation of medicaments for treating vaginitis, pelvic inflammatory disease, cervicitis, adnexitis, and vulvitis.

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