Saussurea involucrate patch for preventing and treating cold in uterus, dysmenorrhea and gynecological reproductive inflammation and preparation method of saussurea involucrate patch

Abstract

The invention provides a saussurea involucrate patch for preventing and treating cold in uterus, dysmenorrhea and gynecological reproductive inflammation and a preparation method of the saussurea involucrate patch, and belongs to the technical field of gynecological nursing. The saussurea involucrate patch is prepared from 14 traditional Chinese medicine active ingredients including 10-25 parts of saussurea involucrate extract, 5-20 parts of folium artemisiae argyi extract, 1-5 parts of saffron crocus and the like, and an azone transdermal enhancer and a nano-scale montmorillonite adsorbent are supplemented. The preparation process adopts a differential extraction technology, firstly, medicinal materials such as astragalus membranaceus are crushed and then subjected to water extraction and alcohol precipitation, and then the medicinal materials are mixed with volatile components such as saffron crocus to prepare the core body. Animal experiments show that the saussurea involucrata paste can significantly reduce the inflammatory factor level and shorten the duration of dysmenorrhea, and has no skin irritation reaction. The traditional Chinese medicine composition achieves the effects of warming channels, dispelling cold, resisting inflammation and inhibiting bacteria through the synergistic effect of multiple components, and is suitable for conditioning cold in uterus and adjuvant therapy of gynecological inflammation.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a snow lotus patch for the prevention and treatment of uterine cold, dysmenorrhea and gynecological reproductive inflammation, and its preparation method. Background Technology

[0002] Cold uterus, dysmenorrhea, and gynecological reproductive system inflammation are common women's health problems in clinical practice, with high incidence and easy recurrence, seriously affecting patients' quality of life. Traditional Chinese medicine theory believes that their pathogenesis is mostly due to deficiency of the root and excess of the branch, and a mixture of cold and heat. The core of "cold uterus" lies in insufficient kidney yang, resulting in the uterus losing its warmth, and the stagnation of cold and dampness, often leading to delayed menstruation, lower abdominal pain during menstruation, pain relieved by warmth, and aversion to cold and cold limbs. As for common reproductive tract inflammations in modern women, such as chronic pelvic inflammatory disease and vaginitis, traditional Chinese medicine often diagnoses them as damp-heat descending downward, obstructing the flow of qi, and over time forming blood stasis, creating a complex pathogenesis of "damp-heat stagnation," manifested as abnormal vaginal discharge, lower abdominal pain or burning pain, and soreness and distension of the lumbosacral region.

[0003] In treatment, modern medicine primarily employs hormone cycle therapy, antibiotics, or nonsteroidal anti-inflammatory drugs (NSAIDs). While these can alleviate symptoms in the short term, long-term use can easily lead to drug resistance, gastrointestinal reactions, and endocrine disorders. Traditional Chinese medicine (TCM) possesses unique advantages in this field, characterized by holistic regulation and fewer side effects. External therapies, in particular, allow medication to directly target the affected area, avoiding the first-pass effect in the liver, and are increasingly favored. However, existing external TCM patches often use single-effect formulas, targeting either dysmenorrhea due to cold in the uterus or gynecological inflammation only, making it difficult to simultaneously address the treatment needs of two conditions. Furthermore, some patches suffer from insufficient extraction of active ingredients, poor transdermal absorption, short duration of efficacy, and inadequate antibacterial effects. Additionally, their complex manufacturing processes result in poor product stability and significant batch-to-batch variations, failing to meet clinical needs. Summary of the Invention

[0004] In view of this, the present invention proposes a snow lotus patch for the prevention and treatment of uterine cold, dysmenorrhea and gynecological reproductive inflammation and its preparation method, thereby solving the above problems.

[0005] The technical solution of the present invention is implemented as follows: a composition for preventing and treating uterine cold, dysmenorrhea and gynecological reproductive inflammation, comprising the following raw materials in parts by weight: 10-25 parts of Tian Shan snow lotus extract, 5-20 parts of Artemisia argyi extract, 1-5 parts of saffron, 8-20 parts of Kochia scoparia extract, 8-20 parts of Cnidium monnieri extract, 5-15 parts of Sophora flavescens extract, 5-12 parts of Dictamnus dasycarpus root bark extract, 5-15 parts of Smilax glabra extract, 5-15 parts of Leonurus japonicus extract, 5-18 parts of Astragalus membranaceus extract, 5-15 parts of Paeonia lactiflora extract, 3-10 parts of Lithospermum erythrorhizon extract, 3-10 parts of Stemona japonica extract, and 1-5 parts of borneol.

[0006] Preferably, the composition comprises the following raw materials in parts by weight: 18 parts of Tian Shan snow lotus extract, 15 parts of Artemisia argyi extract, 3 parts of saffron, 15 parts of Kochia scoparia extract, 12 parts of Cnidium monnieri extract, 10 parts of Sophora flavescens extract, 8 parts of Dictamnus dasycarpus root bark extract, 10 parts of Smilax glabra extract, 10 parts of Leonurus japonicus extract, 12 parts of Astragalus membranaceus extract, 10 parts of Paeonia lactiflora extract, 8 parts of Lithospermum erythrorhizon extract, 8 parts of Stemona japonica extract, and 3 parts of borneol.

[0007] Preferably, the composition further includes a transdermal penetration enhancer and a physical adsorbent. The transdermal penetration enhancer is azone or acetone, added at an amount of 0.5-2 parts by weight; the physical adsorbent is nano-sized montmorillonite or calcined oyster shell powder, added at an amount of 1-8 parts by weight. The transdermal penetration enhancer can significantly improve the transdermal absorption rate of the active ingredient. Azone, as a commonly used chemical penetration enhancer, can promote the penetration and absorption of the active ingredient by altering the lipid arrangement structure of the stratum corneum and increasing the water content of the stratum corneum, with a penetration-enhancing effect similar to that of eugenol, a natural Chinese medicine penetration enhancer. The physical adsorbent can adsorb secretions and harmful substances at the lesion site, while enhancing the stability of the composition. Nano-sized montmorillonite has excellent adsorption properties, adsorbing harmful bacteria and their toxins through electrostatic attraction without affecting the growth of beneficial bacteria. It also has good water absorption and swelling properties, improving the fit and comfort of the patch.

[0008] The present invention provides a method for preparing the above composition, comprising the following steps: S1. Raw material extraction: (1) Take Astragalus membranaceus, Smilax glabra, Paeonia lactiflora and Stemona japonica and grind them into powder to 80-100 mesh sieve. Add purified water at a material-to-liquid ratio of 1:8-12 (g / ml). Soak for 0.5-1 hour, then decoct 2-3 times, 1-2 hours each time. Combine the decoctions, filter, and concentrate the filtrate at 60-70℃ under vacuum to a thick paste with a relative density of 1.10-1.25 at 50℃. Dry it into dry powder to obtain Astragalus membranaceus extract, Smilax glabra extract, Paeonia lactiflora extract and Stemona japonica extract, respectively. (2) Take Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Lithospermum erythrorhizon, Leonurus japonicus and Artemisia argyi raw materials and crush them to an 80-100 mesh sieve. Add 60-85% v / v ethanol solution at a material-to-liquid ratio of 1:6-10 (g / ml). Extract by hot reflux at 70-80℃ 2-3 times, 1.5-2 hours each time. Combine the extracts, filter, recover the ethanol from the filtrate, concentrate into a thick paste, and dry into a dry powder. (3) Grind saffron into 40-60 mesh, mix it with 65-75% v / v ethanol solution at 4-10℃ at a material-to-liquid ratio of 1:8-10 (g / ml), and extract it 1-3 times in pulse mode at 45±2℃ using multi-frequency composite ultrasound, each time for 35-45 min. Combine the extracts and concentrate them to 1 / 10 of the original volume under the conditions of pressure -0.08MPa and temperature ≤50℃, in the dark, to obtain saffron extract with a relative density of about 1.10-1.15 (50℃), and dry it into dry powder. Ultrasonic extraction can destroy the cells of medicinal materials through cavitation effect and stirring action, shorten the extraction time and improve the extraction rate. Moreover, the low extraction temperature can effectively protect the heat-labile active ingredients such as crocin. (4) Place the coarse powder of Tianshan snow lotus in an extraction vessel, introduce CO2 at a flow rate of 20-30L / h, extract for 1.5-3 hours, analyze and collect the extract to obtain Tianshan snow lotus extract; (5) Grind the borneol into ultrafine powder at 4-8℃ and pass it through a 3250-350 mesh sieve for later use; S2. Mixing: Mix the extracts obtained above at a speed of 10-15 rpm for 15-20 minutes to make them initially uniform. Add borneol powder, transdermal penetration enhancer and physical adsorbent, adjust the speed to 18-25 rpm, and continue mixing for 30-45 minutes. Discharge to obtain the composition.

[0009] Preferably, in step S1 (3), the main frequency of the multi-frequency composite ultrasound is 25-30kHz, the auxiliary frequency is 35-45kHz, and the power density is 0.6-0.8W / cm². 3 The pulse mode operates on a 2-second ultrasound cycle followed by a 1-second interval. This parameter setting further enhances the extraction rate of saffron's active ingredients, reduces their degradation, and ensures the extract's efficacy.

[0010] Preferably, in step S1 (4), the extraction pressure is 25-35 MPa and the temperature is 40-50℃. These extraction parameters allow the fat-soluble active ingredients in Tian Shan snow lotus to be fully released, increasing the extraction rate by more than 30% compared to conventional extraction processes, and effectively preserving the activity of the active ingredients.

[0011] This invention provides a snow lotus patch for the prevention and treatment of uterine cold, dysmenorrhea, and gynecological reproductive inflammation. It comprises, from the outside to the inside, a backing layer, a medicated gel layer, and an anti-adhesive layer. The medicated gel layer is made by mixing the above composition with a hydrophilic gel matrix.

[0012] Preferably, the backing layer is a non-woven fabric or a pure cotton spunlace fabric, the anti-adhesive layer is a glassine release paper or a PET release film, and the hydrophilic gel matrix is ​​one or more of carbomer, hydroxypropyl cellulose, and sodium alginate.

[0013] This invention provides a method for preparing the above-mentioned snow lotus patch, comprising the following steps: a. The composition is homogenized with a hydrophilic gel matrix, a humectant, and purified water under vacuum conditions, and the pH value is adjusted to 5.0-6.0 to obtain a drug-containing gel; preferably, the humectant is glycerol or propylene glycol, and the mass-volume ratio (g / mL) of the composition, hydrophilic gel matrix, humectant, and purified water is (2.5-4):(3-5):(1.5-2.5):100; b. Apply the medicated gel evenly to the backing layer at a coating amount of 0.08-0.12 g / cm³. 2 A non-stick layer is applied to form a composite patch; c. The composite patch is dried and shaped at low temperature, controlling the moisture content of the gel layer to 18-28%, then cut and sterilized by irradiation with a dose of 8-15 kGy for 5-8 minutes. The preferred drying temperature is 35-45℃.

[0014] Compared with the prior art, the beneficial effects of the present invention are: The composition of this invention uses Tian Shan snow lotus and mugwort to warm the meridians and dispel cold as the principal ingredients, Kochia scoparia, Cnidium monnieri, and Sophora flavescens to clear heat and dry dampness as the assistant ingredients, Astragalus membranaceus, Leonurus japonicus, and Paeonia lactiflora to invigorate blood and nourish blood as the adjuvant ingredients, and borneol to promote penetration as the guiding ingredient. The combined effects of these herbs are to warm the meridians and dispel cold, clear heat and remove dampness, invigorate blood and remove blood stasis, regulate menstruation and relieve pain. It is effective in treating the common pathogenesis of cold uterus, dysmenorrhea and gynecological inflammation, which involves a mixture of cold and heat and deficiency and excess, thus achieving both symptomatic and root-cause treatment.

[0015] Differentiated optimal extraction processes were adopted to address the characteristics of the active ingredients in different medicinal materials. In particular, low-temperature ultrasound-assisted extraction with specific parameters was used for saffron, and supercritical CO2 extraction was used for snow lotus. These methods maximized the retention and enrichment of heat-sensitive and fat-soluble active ingredients, improved extraction efficiency and product quality, and laid the material foundation for the product's excellent therapeutic effects.

[0016] This invention combines the above composition with a hydrophilic gel matrix to form a modern patch dosage form that promotes drug dissolution and transdermal delivery, enabling continuous drug administration and convenient use. Detailed Implementation

[0017] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0018] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0019] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0020] Example 1 The composition formula includes: 10g of Tian Shan snow lotus extract, 5g of Artemisia argyi extract, 1g of saffron, 8g of Kochia scoparia extract, 8g of Cnidium monnieri extract, 5g of Sophora flavescens extract, 5g of Dictamnus dasycarpus root bark extract, 5g of Smilax glabra extract, 5g of Leonurus japonicus extract, 5g of Astragalus membranaceus extract, 5g of Paeonia lactiflora extract, 3g of Lithospermum erythrorhizon extract, 3g of Stemona japonica extract, 1g of borneol, 0.5g of transdermal penetration enhancer aminoketone, and 1g of nano-sized montmorillonite as a physical adsorbent.

[0021] The method for preparing the above composition includes the following steps: S1. Raw material extraction: (1) Take Astragalus membranaceus, Smilax glabra, Paeonia lactiflora and Stemona japonica and grind them into 80 mesh sieves. Add purified water at a material-to-liquid ratio of 1:8 (g / ml). Soak for 0.5 hours, decoct twice, 1 hour each time, combine the decoctions, filter, concentrate the filtrate at 60℃ and vacuum conditions to a thick paste with a relative density of 1.10 at 50℃, dry it into dry powder, and obtain Astragalus membranaceus extract, Smilax glabra extract, Paeonia lactiflora extract and Stemona japonica extract respectively. (2) Take Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Lithospermum erythrorhizon, Leonurus japonicus and Artemisia argyi raw materials and crush them to an 80-mesh sieve. Add 60% v / v ethanol solution at a material-to-liquid ratio of 1:6 (g / ml). Extract twice at 70℃ for 1.5 hours each time. Combine the extracts, filter, recover the ethanol from the filtrate, concentrate it into a thick paste, and dry it into a dry powder. (3) Grind saffron into 40 mesh, mix it with 65% v / v ethanol solution at 4℃ at a material-to-liquid ratio of 1:8 (g / ml), and use multi-frequency composite ultrasound (main frequency 25kHz, auxiliary frequency 35kHz, power density 0.6W / cm³). 3 The pulse mode working cycle is 2 seconds of ultrasound followed by 1 second of intermittent extraction. Extraction is performed once at 43℃ for 35 minutes each time. The extracts are combined and concentrated to 1 / 10 of the original volume under the conditions of -0.08MPa pressure and 45℃ in the dark to obtain saffron extract with a relative density of 1.10 (50℃). The extract is then dried into powder. (4) Place the coarse powder of Tianshan snow lotus in an extraction vessel, introduce CO2 at a flow rate of 20L / h, extract at an extraction pressure of 25MPa and a temperature of 40℃ for 1.5 hours, and collect the extract to obtain Tianshan snow lotus extract. (5) Grind the borneol into ultrafine powder at 4°C and pass it through a 325-mesh sieve for later use; S2. Mixing: Mix the above-obtained extracts at 10 rpm for 15 minutes according to the prescription to make them initially uniform. Add borneol powder, amino ketone and nano-sized montmorillonite, adjust the speed to 18 rpm, and continue mixing for 30 minutes. Discharge to obtain the composition.

[0022] Example 2 The composition formula includes: 25g of Tian Shan snow lotus extract, 20g of Artemisia argyi extract, 5g of saffron, 20g of Kochia scoparia extract, 20g of Cnidium monnieri extract, 15g of Sophora flavescens extract, 12g of Dictamnus dasycarpus root bark extract, 15g of Smilax glabra extract, 15g of Leonurus japonicus extract, 18g of Astragalus membranaceus extract, 15g of Paeonia lactiflora extract, 10g of Lithospermum erythrorhizon extract, 10g of Stemona japonica extract, 5g of borneol, 2g of transdermal penetration enhancer azone, and 8g of nano-sized montmorillonite as a physical adsorbent.

[0023] The method for preparing the above composition includes the following steps: S1. Raw material extraction: (1) Take Astragalus membranaceus, Smilax glabra, Paeonia lactiflora and Stemona japonica and grind them into 100 mesh sieves. Add purified water at a material-to-liquid ratio of 1:12 (g / ml). After soaking for 1 hour, decoct 3 times for 2 hours each time. Combine the decoctions, filter, and concentrate the filtrate at 70℃ and vacuum conditions to a thick paste with a relative density of 1.25 at 50℃. Dry it into dry powder to obtain Astragalus membranaceus extract, Smilax glabra extract, Paeonia lactiflora extract and Stemona japonica extract respectively. (2) Take Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Lithospermum erythrorhizon, Leonurus japonicus and Artemisia argyi respectively and crush them to a 100-mesh sieve. Add 85% v / v ethanol solution at a material-to-liquid ratio of 1:10 (g / ml). Extract by hot reflux at 80℃ 3 times, 2 hours each time. Combine the extracts, filter, recover the ethanol from the filtrate and concentrate it into a thick paste, and dry it into a dry powder. (3) Grind saffron into 60 mesh, mix it with 75% v / v ethanol solution at 10℃ at a material-to-liquid ratio of 1:10 (g / ml), and use multi-frequency composite ultrasound (main frequency 30kHz, auxiliary frequency 45kHz, power density 0.8 W / cm³). 3 The pulse mode working cycle is 2 seconds of ultrasound followed by 1 second of intermittent extraction. The extract was extracted 3 times at 47℃ for 45 minutes each time. The extracts were combined and concentrated to 1 / 10 of the original volume under the conditions of -0.08 MPa pressure and 50℃ temperature, in the dark, to obtain saffron extract with a relative density of 1.15 (50℃). The extract was then dried into powder. (4) Place the coarse powder of Tianshan snow lotus in an extraction vessel, introduce CO2 at a flow rate of 30 L / h, extract at an extraction pressure of 35 MPa and a temperature of 50℃ for 3 hours, and collect the extract to obtain Tianshan snow lotus extract. (5) Grind the borneol into ultrafine powder at 8°C and pass it through a 350-mesh sieve for later use; S2. Mixing: Mix the above-obtained extracts at 15 rpm for 20 minutes according to the prescription to make them initially uniform. Add borneol powder, azone and nano-sized montmorillonite, adjust the speed to 25 rpm, and continue mixing for 45 minutes. Discharge to obtain the composition.

[0024] Example 3 The composition formula includes: 18g of Tian Shan snow lotus extract, 15g of Artemisia argyi extract, 3g of saffron, 15g of Kochia scoparia extract, 12g of Cnidium monnieri extract, 10g of Sophora flavescens extract, 8g of Dictamnus dasycarpus root bark extract, 10g of Smilax glabra extract, 10g of Leonurus japonicus extract, 12g of Astragalus membranaceus extract, 10g of Paeonia lactiflora extract, 8g of Lithospermum erythrorhizon extract, 8g of Stemona japonica extract, 3g of borneol, 1.2g of transdermal penetration enhancer azone, and 4g of calcined oyster shell powder as a physical adsorbent.

[0025] The method for preparing the above composition includes the following steps: S1. Raw material extraction: (1) Take Astragalus membranaceus, Smilax glabra, Paeonia lactiflora and Stemona japonica and grind them into 90 mesh sieves. Add purified water at a material-to-liquid ratio of 1:10 (g / ml). Soak for 0.8 hours, then decoct twice for 1.5 hours each time. Combine the decoctions, filter, and concentrate the filtrate at 65℃ and vacuum conditions to a thick paste with a relative density of 1.18 at 50℃. Dry it into dry powder to obtain Astragalus membranaceus extract, Smilax glabra extract, Paeonia lactiflora extract and Stemona japonica extract, respectively. (2) Take Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Lithospermum erythrorhizon, Leonurus japonicus and Artemisia argyi raw materials and crush them to a 90-mesh sieve. Add 75% v / v ethanol solution at a material-to-liquid ratio of 1:8 (g / ml). Extract twice at 78℃ for 1.8 hours each time. Combine the extracts, filter, recover the ethanol from the filtrate, concentrate it into a thick paste, and dry it into a dry powder. (3) Grind saffron into 50 mesh, mix it with 70% v / v ethanol solution at 7℃ at a material-to-liquid ratio of 1:9 (g / ml), and use multi-frequency composite ultrasound (main frequency 28kHz, auxiliary frequency 40kHz, power density 0.7 W / cm³). 3 The pulse mode working cycle is 2 seconds of ultrasound followed by 1 second of intermittent extraction. The extract was extracted twice at 45℃ for 40 minutes each time. The extracts were combined and concentrated to 1 / 10 of the original volume under the conditions of -0.08 MPa pressure and 48℃ temperature, in the dark, to obtain saffron extract with a relative density of 1.12 (50℃). The extract was then dried into powder. (4) Place the coarse powder of Tianshan snow lotus in an extraction vessel, introduce CO2 at a flow rate of 25L / h, extract at an extraction pressure of 30MPa and a temperature of 45℃ for 2 hours, and collect the extract to obtain Tianshan snow lotus extract. (5) Grind the borneol into ultrafine powder at 6°C and pass it through a 340-mesh sieve for later use; S2. Mixing: Mix the above-obtained extracts at 12 rpm for 18 minutes according to the prescription to make them initially uniform. Add borneol powder, azone and calcined oyster powder, adjust the speed to 22 rpm, and continue mixing for 38 minutes. Discharge to obtain the composition.

[0026] The snow lotus patch is prepared by the following steps: (The composition of Examples 1-3 above is used to prepare the patch.) a. The composition was homogenized with hydroxypropyl cellulose, propylene glycol, and purified water at a mass-volume ratio of 3.2:4:2:100 (g / mL) under vacuum conditions, and the pH was adjusted to 5.5 to obtain a drug-containing gel. b. Dissolve the medicated gel at 0.10 g / cm³. 2 The coating amount is evenly applied to the pure cotton spunlace fabric backing layer, and covered with a PET release film anti-stick layer to form a composite patch. c. The composite patch is dried and shaped at 40°C, and the water content of the gel layer is controlled at 23%. Then it is cut and sterilized by irradiation with a dose of 12kGy for 6 minutes to obtain the snow lotus patch.

[0027] Comparative Example 1 The difference between this comparative example and Example 3 is that it lacks extracts of Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Smilax glabra, and Stemona japonica; the extraction, mixing, and preparation steps and parameters of the remaining components and compositions are the same as in Example 3.

[0028] Comparative Example 2 The difference between this comparative example and Example 3 is that it lacks extracts of Tian Shan snow lotus, mugwort, and saffron. The extraction, mixing, and preparation steps and parameters of the remaining components and compositions are the same as in Example 3.

[0029] Comparative Example 3 The difference between this comparative example and Example 3 is that it lacks extracts of Leonurus japonicus, Astragalus membranaceus, Paeonia lactiflora, and Lithospermum erythrorhizon. The extraction, mixing, and preparation steps and parameters of the remaining components and the composition are the same as those in Example 3.

[0030] Comparative Example 4 The difference between this comparative example and Example 3 is that the medicinal materials were mixed and added to a 75% v / v ethanol solution at a material-to-liquid ratio of 1:8 (g / ml) and extracted at 78°C for 2 hours.

[0031] Experimental example: Pharmacodynamic experiment This experiment verified the efficacy of the snow lotus patch prepared in Examples 1-3 and Comparative Examples 1-4 on uterine cold, dysmenorrhea, and gynecological reproductive inflammation through animal experiments, clarified the synergistic effect of the formula and the rationality of the process, and strictly followed the "Technical Guidelines for Pharmacodynamic Research of Traditional Chinese Medicine" to ensure that the experimental results are scientific, reliable and reproducible.

[0032] I. Experimental Materials and Instruments 1. Test samples: Snow lotus patches prepared in Examples 1-3 and Comparative Examples 1-4 (cut into 1cm×1cm sizes for later use); positive control drugs (cold uterus and dysmenorrhea: Artemisia argyi warming uterus patch; gynecological inflammation: metronidazole suppositories).

[0033] 2. Experimental animals: SPF-grade female SD rats (weight 200±20g, 6-8 weeks old) and SPF-grade female Kunming mice (weight 20±2g, 6-8 weeks old) were purchased from a regular experimental animal center and were acclimatized for 3 days (temperature 22±2℃, humidity 50±5%, free access to food and water) before being used in the experiment.

[0034] 3. Reagents and instruments: Oxytocin (to induce dysmenorrhea model), estrogen (to induce uterine cold model), Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC 29213), Candida albicans (ATCC 10231), physiological saline, formaldehyde solution, optical microscope, electronic balance, centrifuge, ELISA reader, etc.

[0035] II. Establishment of Animal Models This experiment was divided into three efficacy modules: a cold uterus model, a dysmenorrhea model, and a gynecological inflammation (vaginitis) model, which correspond to the core indications of the snow lotus patch of this invention. The model establishment method is as follows: 1. Cold Uterus Model (Rats): Female SD rats were injected subcutaneously with estrogen (0.5 mg / kg) daily for 7 consecutive days. At the same time, the rats were kept in a low temperature environment of 4±1℃ (4 hours per day). On the 8th day, the rectal temperature and uterine index of the rats were measured. The model was considered to be successfully established if the rectal temperature decreased by ≥0.5℃ and the uterine index increased significantly compared with normal rats.

[0036] 2. Dysmenorrhea model (mice): Female Kunming mice were administered estrogen (0.2 mg / mouse) by gavage for 3 consecutive days. On the 4th day, oxytocin (2 U / mouse) was injected intraperitoneally. The writhing response (abdominal contraction, hind limb extension, and buttock elevation) of the mice was observed. The model was considered to be successfully established when there was an obvious writhing response (≥10 writhing times within 10 minutes).

[0037] 3. Vaginitis model (rat): Female SD rats were anesthetized, and a mixed bacterial solution (Escherichia coli, Staphylococcus aureus, and Candida albicans, each at a concentration of 1×10⁻⁶) was collected using a sterile cotton swab. 8 The CFU / mL solution was slowly inserted deep into the rat's vagina (about 1 cm) once a day for 3 consecutive days. On the 4th day, a smear of vaginal secretions was taken, and a large number of pathogenic bacteria were observed under a microscope, indicating that the model was successfully established.

[0038] III. Experimental Grouping and Administration Methods Each model was set up with 7 groups, with 10 animals in each group. The grouping and administration methods are as follows: the administration period for all groups is 7 days, once a day: 1. Blank control group: Normal animals, treated with physiological saline only, without any drug administration; 2. Model control group: Model animals, treated only with physiological saline without any drug administration; 3. Positive control group: Model animals, the cold uterus / dysmenorrhea model was given Ai Fu warming uterus patch (applied to the lower abdomen), and the vaginitis model was given metronidazole suppositories (vaginal administration, 0.1g / animal). 4. Example 1 group: model animals, with the snow lotus patch prepared in Example 1 applied (applied to the lower abdomen / perineum, corresponding to the model). 5. Example 2 group: model animals, with the snow lotus patch prepared in Example 2 applied (applied to the lower abdomen / perineum, corresponding to the model); 6. Example 3 group: model animals, with the snow lotus patch prepared in Example 3 applied (applied to the lower abdomen / perineum, corresponding to the model); 7. Comparative Examples 1-4: Model animals, each of which was covered with snow lotus patches prepared in Comparative Examples 1-4 (applied to the lower abdomen / perineum, corresponding to the model).

[0039] Note: Ensure the patch adheres tightly to the skin / mucous membrane during administration to prevent it from falling off; clean the rat's vagina with physiological saline before administration to the vaginitis model.

[0040] IV. Testing Indicators and Judgment Criteria Seven days after administration, relevant efficacy indicators of each model were measured to objectively evaluate the efficacy of the Snow Lotus Patch, as detailed below: (I) Efficacy testing of drugs in the model of uterine cold 1. Detection indicators: Rectal temperature: The rectal temperature of rats was measured using an electronic thermometer (inserted 1.5 cm into the anus and left in place for 3 min); Uterine index: Sacrifice the rats, remove the uterus, weigh the uterus using an electronic balance, and calculate the uterine index (uterine index = uterine weight / rat body weight × 100%). Uterine histopathology: Uterine tissue was taken, fixed in formaldehyde, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and the morphology of the uterine mucosa was observed under an optical microscope (whether there was edema, congestion, or inflammatory cell infiltration).

[0041] 2. Judgment criteria: Effective (rectal temperature returns to the normal range, uterine index significantly decreases, uterine mucosal edema and congestion are significantly relieved, and inflammatory cell infiltration is reduced); Ineffective (rectal temperature and uterine index do not improve significantly, and uterine mucosal lesions are not relieved).

[0042] (II) Detection of the efficacy of drugs in the dysmenorrhea model 1. Detection indicators: Number of writhing events: Seven days after administration, oxytocin (2U / mouse) was injected intraperitoneally, and the number of writhing events in mice within 10 minutes was recorded. Writhing latency: The time when mice first exhibited a writhing response after injection of oxytocin was recorded. Serum prostaglandin E2 (PGE2) content: Mice were sacrificed, serum was collected, and PGE2 content was detected by enzyme-linked immunosorbent assay (ELISA) (the higher the PGE2 content, the more severe the dysmenorrhea symptoms).

[0043] 2. Judgment criteria: Effective (the number of writhing movements is reduced by ≥30% compared with the model control group, the writhing latency is prolonged by ≥50%, and the serum PGE2 level is significantly reduced); Ineffective (no significant improvement in the number of writhing movements, latency, or PGE2 level).

[0044] (III) Drug efficacy testing in a vaginitis model 1. Detection indicators: Pathogenic bacteria count in vaginal secretions: Vaginal secretions from rats were diluted, inoculated into a culture medium, and cultured for 24 hours. The number of pathogenic bacteria colonies was then counted. Vaginal histopathology: Vaginal tissue was taken, fixed in formaldehyde, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and the morphology of the vaginal mucosa (whether there was erosion, hyperemia, or inflammatory cell infiltration) was observed under a light microscope. Serum inflammatory factor (TNF-α, IL-6) levels: Rats were sacrificed, serum was collected, and TNF-α and IL-6 levels were detected by enzyme-linked immunosorbent assay (ELISA) (the higher the level, the more severe the inflammation).

[0045] 2. Judgment criteria: Effective (the number of pathogenic bacteria colonies decreased by ≥90% compared with the model control group, vaginal mucosal erosion and congestion were significantly relieved, inflammatory cell infiltration was reduced, and serum TNF-α and IL-6 levels were significantly reduced); Ineffective (the number of pathogenic bacteria colonies and the content of inflammatory factors did not improve significantly, and vaginal mucosal lesions were not relieved).

[0046] Effectiveness (%) = (Number of effective animals / Total number of animals in the group) × 100% V. Test Results Table 1. Efficacy results of the drug in the model of uterine cold

[0047] Table 2. Efficacy results of the dysmenorrhea model

[0048] Table 3. Efficacy results of the vaginitis model

[0049] Animal experiments have confirmed that the snow lotus patch prepared in Examples 1-3 of this invention has significant therapeutic effects on uterine cold, dysmenorrhea and gynecological reproductive inflammation. The synergistic effect of each component in the formula is obvious, and the differentiated extraction process can ensure efficacy and high safety.

[0050] The above embodiments and comparative examples underwent safety testing. The test results showed that none of the rats exhibited irritation reactions such as redness, swelling, itching, or ulceration on their backs, indicating that the snow lotus patch of the present invention has high safety and no obvious skin irritation.

[0051] 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 composition for the prevention and treatment of uterine coldness, dysmenorrhea, and gynecological reproductive inflammation, characterized in that, The ingredients include the following parts by weight: 10-25 parts of Tian Shan snow lotus extract, 5-20 parts of Artemisia argyi extract, 1-5 parts of saffron, 8-20 parts of Kochia scoparia extract, 8-20 parts of Cnidium monnieri extract, 5-15 parts of Sophora flavescens extract, 5-12 parts of Dictamnus dasycarpus root bark extract, 5-15 parts of Smilax glabra extract, 5-15 parts of Leonurus japonicus extract, 5-18 parts of Astragalus membranaceus extract, 5-15 parts of Paeonia lactiflora extract, 3-10 parts of Lithospermum erythrorhizon extract, 3-10 parts of Stemona japonica extract, and 1-5 parts of borneol.

2. The composition according to claim 1, characterized in that, The ingredients include the following parts by weight: 18 parts of Tian Shan snow lotus extract, 15 parts of Artemisia argyi extract, 3 parts of saffron, 15 parts of Kochia scoparia extract, 12 parts of Cnidium monnieri extract, 10 parts of Sophora flavescens extract, 8 parts of Dictamnus dasycarpus root bark extract, 10 parts of Smilax glabra extract, 10 parts of Leonurus japonicus extract, 12 parts of Astragalus membranaceus extract, 10 parts of Paeonia lactiflora extract, 8 parts of Lithospermum erythrorhizon extract, 8 parts of Stemona japonica extract, and 3 parts of borneol.

3. The composition according to claim 1, characterized in that, It also includes a transdermal penetration enhancer and a physical adsorbent, wherein the transdermal penetration enhancer is ammonia ketone or azone, and the amount added is 0.5-2 parts by weight; the physical adsorbent is nano-sized montmorillonite or calcined oyster powder, and the amount added is 1-8 parts by weight.

4. The method for preparing the composition according to claim 3, characterized in that, Includes the following steps: S1. Raw material extraction: (1) Take Astragalus membranaceus, Smilax glabra, Paeonia lactiflora and Stemona japonica and grind them into powder to 80-100 mesh sieve. Add purified water at a material-to-liquid ratio of 1:8-12 (g / ml). Soak for 0.5-1 hour, then decoct 2-3 times, 1-2 hours each time. Combine the decoctions, filter, and concentrate the filtrate at 60-70℃ under vacuum to a thick paste with a relative density of 1.10-1.25 at 50℃. Dry it into dry powder to obtain Astragalus membranaceus extract, Smilax glabra extract, Paeonia lactiflora extract and Stemona japonica extract, respectively. (2) Take Kochia scoparia, Cnidium monnieri, Sophora flavescens, Dictamnus dasycarpus, Lithospermum erythrorhizon, Leonurus japonicus and Artemisia argyi raw materials and crush them to an 80-100 mesh sieve. Add 60-85% v / v ethanol solution at a material-to-liquid ratio of 1:6-10 (g / ml). Extract by hot reflux at 70-80℃ 2-3 times, 1.5-2 hours each time. Combine the extracts, filter, recover the ethanol from the filtrate, concentrate into a thick paste, and dry into a dry powder. (3) Grind saffron into 40-60 mesh, mix it with 65-75% v / v ethanol solution at 4-10℃ at a material-to-liquid ratio of 1:8-10 (g / ml), and extract it 1-3 times in pulse mode at 45±2℃ using multi-frequency composite ultrasound, each time for 35-45 min. Combine the extracts and concentrate them to 1 / 10 of the original volume under the conditions of pressure -0.08MPa and temperature ≤50℃, in the dark, to obtain saffron extract with a relative density of about 1.10-1.15 (50℃), and dry it into dry powder. (4) Place the coarse powder of Tianshan snow lotus in an extraction vessel, introduce CO2 at a flow rate of 20-30L / h, extract for 1.5-3 hours, analyze and collect the extract to obtain Tianshan snow lotus extract; (5) Grind the borneol into ultrafine powder at 4-8℃ and pass it through a 3250-350 mesh sieve for later use; S2. Mixing: Mix the extracts obtained above at a speed of 10-15 rpm for 15-20 minutes to make them initially uniform. Add borneol powder, transdermal penetration enhancer and physical adsorbent, adjust the speed to 18-25 rpm, and continue mixing for 30-45 minutes. Discharge to obtain the composition.

5. The preparation method according to claim 4, characterized in that, In step S1(3), the main frequency of the multi-frequency composite ultrasound is 25-30kHz, the auxiliary frequency is 35-45kHz, and the power density is 0.6-0.8W / cm². 3 The pulse mode has a working cycle of 2 seconds of ultrasound followed by a 1-second interval.

6. The preparation method according to claim 4, characterized in that, In step S1 (4), the extraction pressure is 25-35 MPa and the temperature is 40-50℃.

7. A snow lotus patch for the prevention and treatment of uterine coldness, dysmenorrhea, and gynecological reproductive inflammation, characterized in that... From the outside to the inside, it includes a backing layer, a drug-containing gel layer, and an anti-adhesive layer, wherein the drug-containing gel layer is made by mixing the composition of any one of claims 1-6 with a hydrophilic gel matrix.

8. The snow lotus patch as described in claim 7, characterized in that, The backing layer is non-woven fabric or pure cotton spunlace fabric, the anti-adhesive layer is glassine release paper or PET release film, and the hydrophilic gel matrix is ​​one or more combinations of carbomer, hydroxypropyl cellulose, and sodium alginate.

9. The method for preparing snow lotus patch as described in claim 7 or 8, characterized in that, Includes the following steps: a. The composition is homogenized with a hydrophilic gel matrix, a humectant, and purified water under vacuum conditions, and the pH value is adjusted to 5.0-6.0 to obtain a drug-containing gel; b. Apply the drug-containing gel evenly onto the backing layer, cover it with an anti-adhesive layer, and form a composite patch; c. The composite patch is dried and shaped at low temperature, and the water content of the gel layer is controlled at 18-28%. Then it is cut and sterilized by irradiation with a dose of 8-15 kGy for 5-8 minutes to obtain the final product.

10. The preparation method according to claim 9, characterized in that, In step a, the moisturizer is glycerin or propylene glycol, and the mass-to-volume ratio (g / mL) of the composition, hydrophilic gel matrix, moisturizer, and purified water is (2.5-4):(3-5):(1.5-2.5):100; in step b, the coating amount is 0.08-0.12 g / cm³. 2 In step c, the drying temperature is 35-45℃.

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