Melastoma dodecandrum temperature-sensitive in-situ gel as well as preparation method and application thereof

Abstract

The invention discloses melastoma dodecandrum temperature-sensitive in-situ gel as well as a preparation method and application thereof. The melastoma dodecandrum lour temperature-sensitive in-situ gel is prepared from a melastoma dodecandrum lour extract, poloxamer 407, poloxamer 188, isopropyl myristate, tween-80 and glycerin, and the preparation method comprises the following steps: adding purified water into the poloxamer 407 and the poloxamer 188 for swelling, refrigerating and standing until the solution is clear, adding the glycerin, and uniformly mixing to obtain a solution A; dissolving the melastoma dodecandrum lour extract in isopropyl myristate in a water bath, adding Tween-80 to obtain a solution B, and uniformly mixing the solution A and the solution B to obtain the traditional Chinese medicine composition. The melastoma dodecandrum lour temperature-sensitive in-situ gel prepared by the invention meets the physiological environment requirements of vagina, has better overall performance such as loading capacity, gelling temperature, viscosity and the like, and also has good bacteriostatic and anti-inflammatory effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel, a preparation method thereof and an application thereof. Background Art

[0002] Gynecological inflammations (such as vaginitis and cervicitis) are common clinical diseases, and there is a significant correlation between their chronic persistence and the occurrence and development of cervical cancer. Although the antibiotic suppositories and lotions widely used clinically at present can temporarily relieve symptoms, there are bottleneck problems such as short drug retention time, high drug resistance generation rate and local microenvironment damage. It is worth noting that the incidence of cervical cancer in patients with chronic cervicitis is 5-7 times higher than that in the normal population, suggesting that blocking the inflammation-cancer transformation by regulating the inflammatory microenvironment has important scientific value.

[0003] Melastoma dodecandrum Lour. is a characteristic medicinal material for treating leukorrhea and cervicitis in folk. Modern research reveals that it is rich in flavonoids, tannins and triterpenoid compounds, and has activities such as antibacterial, anti-inflammatory, anti-cancer and hemostasis [1-3] . In an LPS-induced rat model, the water extract of Melastoma dodecandrum Lour. can reduce the expression levels of inflammation-related factors (IL-1β, IL-6 and TNF-α) and oxidative stress-related factors (MDA, SOD and NO), and alleviate LPS-induced inflammation and LPS-induced liver and kidney injuries by regulating mitochondrial apoptosis-related proteins in splenic CD4+ T cells, inhibiting the production of reactive oxygen species and mitochondrial apoptosis [4] .

[0004] Previous studies in our team found that the Melastoma dodecandrum extract (MDE) has effects such as anti-cervicitis, anti-cervical cancer and hemostasis. Currently, research on the transformation from cervicitis to cancer is underway, aiming to prevent cervical cancer by treating cervicitis. Combining with HPLC fingerprinting, it was found that MDE has obvious inhibitory effects on NO and TNF-α, and components such as vitexin, isovitexin and quercetin may be the key pharmacodynamic substances for MDE to exert its anti-inflammatory effect [5-8] , providing a basis for the quality control of Melastoma dodecandrum Lour. medicinal materials, samples, extracts and preparations.

[0005] Temperature-sensitive gel is a kind of polymer material, which can be administered in liquid form at room temperature and form a semi-solid gel after contacting body temperature, prolonging the drug retention time and realizing targeted sustained release [9-12] . At present, there is no report on the combination of Melastoma dodecandrum extract and temperature-sensitive gel.

[0006] Therefore, the research team of this invention used MDE as the model drug and prepared a new thermosensitive drug delivery system with the best mechanical properties and gel temperature from the gel material according to the optimized ratio. The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel prepared in this invention has good loading capacity, gelling temperature, viscosity, antibacterial and anti-inflammatory effects, etc., can prolong the drug retention time and achieve targeted slow release, does not damage the local microenvironment, has practical application value for the prevention and treatment of gynecological diseases, and provides technical support for the high-quality development of Melastoma dodecandrum Lour. resources and the innovation of local drug delivery systems for gynecology. Summary of the Invention

[0007] The purpose of this invention is to provide a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel.

[0008] Another purpose of this invention is to provide a preparation method of a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel.

[0009] Another purpose of this invention is to provide an application of a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel.

[0010] The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel described in this invention is composed of 4 - 8 g of Melastoma dodecandrum Lour. extract, 9.6 - 12.8 g of poloxamer 407, 1.2 - 4.4 g of poloxamer 188, 6 - 14 mL of isopropyl myristate, 6 - 14 mL of Tween-80, and 1 - 3 mL of glycerol.

[0011] Preferably, the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel described in this invention is composed of 5 - 7 g of Melastoma dodecandrum Lour. extract, 10.4 - 12.0 g of poloxamer 407, 2.0 - 3.6 g of poloxamer 188, 10 - 14 mL of isopropyl myristate, 8 - 12 mL of Tween-80, and 2 - 3 mL of glycerol.

[0012] More preferably, the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel described in this invention is composed of 6 g of Melastoma dodecandrum Lour. extract, 11.2 g of poloxamer 407, 2.8 g of poloxamer 188, 12 mL of isopropyl myristate, 10 mL of Tween-80, and 2 mL of glycerol.

[0013] The preparation method of the Melastoma dodecandrum Lour. extract described in this invention is as follows: Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal materials, add 14 times the amount of 95% ethanol, reflux and extract twice, each time for 1 hour, filter, combine the filtrates, concentrate to a relative density of 1.02, add water to the extract to make a suspension, extract the suspension with petroleum ether (1:1) until colorless, take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0014] The preparation method of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention is as follows: Weigh 9.6 - 12.8 g of poloxamer 407 and 1.2 - 4.4 g of poloxamer 188 into a beaker, add 80 - 120 mL of ultrapure water for swelling, place it in a -4°C refrigerator for 20 - 28 h until it becomes clear, add 1 - 3 mL of glycerol, and mix well to obtain Solution A; Take 4 - 8 g of Melastoma dodecandrum Lour. extract, dissolve it in 6 - 14 mL of isopropyl myristate under a water bath at 40 - 60°C, add 6 - 14 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 5 - 20 min until it is uniform, then it is obtained.

[0015] Preferably, the preparation method of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention is as follows: Weigh 10.4 - 12.0 g of poloxamer 407 and 2.0 - 3.6 g of poloxamer 188 into a beaker, add 90 - 110 mL of ultrapure water for swelling, place it in a -4°C refrigerator for 22 - 26 h until it becomes clear, add 2 - 3 mL of glycerol, and mix well to obtain Solution A; Take 5 - 7 g of Melastoma dodecandrum Lour. extract, dissolve it in 10 - 14 mL of isopropyl myristate under a water bath at 45 - 55°C, add 8 - 12 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 10 - 15 min until it is uniform, then it is obtained.

[0016] More preferably, the preparation method of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention is as follows: Weigh 11.2 g of poloxamer 407 and 2.8 g of poloxamer 188 into a beaker, add 100 mL of ultrapure water for swelling, place it in a -4°C refrigerator for 24 h until it becomes clear, add 2 mL of glycerol, and mix well to obtain Solution A; Take 6 g of Melastoma dodecandrum Lour. extract, dissolve it in 12 mL of isopropyl myristate under a water bath at 50°C, add 10 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 10 min until it is uniform, then it is obtained.

[0017] The application of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention in the preparation of drugs for inhibiting and anti-inflammatory of gynecological inflammation.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention meets the requirements of the vaginal physiological environment. Based on single-factor experiments, the optimal formulation of the Melastoma dodecandrum Lour. in-situ gel MD-TSIG is optimized by the response surface method as P407 11.2 g, P188 2.8 g, and IPM 12 g. The average gelation temperature of the prepared MD-TSIG is 33.4°C, within the range of the optimal gelation temperature of 31 - 34°C for temperature-sensitive gels.

[0020] 2. The preparation method of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention is stable and reliable. The experimental data is subjected to quadratic multiple regression fitting analysis. The model P of the regression equation is < 0.01, and the P value of the lack-of-fit term is 0.7022 > 0.05. The model correlation coefficient R 2 = 0.9662, and the adjusted determination coefficient R 2 Adj = 0.9228, indicating that the model has good credibility, there is no lack-of-fit factor and it can fit well with the actual value, and the model can explain the change of the response value. Through the variance analysis of each factor, it is found that the order of the influence of each factor on the gelation temperature is B (P188) > A (P407) > C (IPM), and all three factors have extremely significant effects on the linear effect of the gelation temperature (P < 0.01). In summary, the regression model has good predictability, and the optimized MD-TSIG preparation process conditions are accurate and reliable.

[0021] 3. The present invention screens the optimal solvent for the Melastoma dodecandrum Lour. extract. The results show that the solubility of the Melastoma dodecandrum Lour. extract in olive oil, liquid paraffin, soybean oil, ethyl acetate, and IPM from high to low is: ethyl acetate > IPM > soybean oil > liquid paraffin > olive oil. Although the solubility of ethyl acetate is the highest, IPM is selected as the optimal solvent because of its non-volatility and pharmaceutical-grade compatibility.

[0022] 4. The pH of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel sample of the present invention is 5.29 ± 0.03, which meets the vaginal administration preparation standard (4.5 - 5.5).

[0023] 5. The components of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel prepared by the present invention are stable. The infrared spectrum results show that there is good compatibility between the materials forming the gel and no component change occurs.

[0024] 6. The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention has good antibacterial effects. The in vitro antibacterial experiment results show that MD-TSIG has antibacterial properties against both S. aureus and E. coli, and with the increase of the MD-TSIG concentration, its antibacterial effect becomes more obvious.

[0025] 7. The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel of the present invention has good anti-inflammatory effects. The pharmacodynamic research shows that the uterus of the rats in the model group shows different degrees of inflammatory cell infiltration, mainly lymphocytes, neutrophils, and eosinophils, indicating that an inflammatory reaction occurs in the reproductive tract. The uterine tissue lesions in the MDE administration group are relatively milder than those in the model group, suggesting that MDE has certain anti-inflammatory effects. Description of the Drawings

[0026] Figure 1Results of single-factor investigation on gelation temperature (a: effect of P407 dosage on gelation temperature; b: effect of P188 dosage on gelation temperature; c: effect of IPM dosage on gelation temperature; d: effect of Tween-80 dosage on gelation temperature; e: effect of glycerol dosage on gelation temperature; f: effect of stirring time on gelation temperature);

[0027] Figure 2 Box-Behnken response surface plots (left: plot of the interaction between P188 and P407 on gelation temperature; middle: plot of the interaction between IPM and P407 on gelation temperature; right: plot of the interaction between IPM and P188 on gelation temperature);

[0028] Figure 3 Appearance and microscopic morphology of MD-TSIG (a: appearance at room temperature and 37 °C (left is room temperature, right is 37 °C); b, c: microscopic morphology);

[0029] Figure 4 Rheological test results of MD-TSIG (a: curves of storage modulus G′ and loss modulus G″ versus strain γ*; b: curves of storage modulus G′ and loss modulus G″ versus scanning frequency f; c: curves of storage modulus G′ and loss modulus G″ versus temperature T);

[0030] Figure 5 Infrared spectra of each component of MD-TSIG;

[0031] Figure 6 TG / DTG curve of MD-TSIG;

[0032] Figure 7 Changes in gel viscosity and appearance after dilution with simulated vaginal fluid (a: effect of temperature change on complex viscosity η*; b: appearance shape; c: centrifugation results of MD-TSIG after being placed at 4, 25, and 37 °C for a period of time);

[0033] Figure 8 Antibacterial effect of MD-TSIG (left figure: antibacterial results against S. aureus (from left to right are blank control group, 0.5 g MD-TSIG group, 1 g MD-TSIG group); right figure: antibacterial results against E. coli (from left to right are blank control group, 0.5 g MD-TSIG group, 1 g MD-TSIG group));

[0034] Figure 9 Uterus diagrams of rats in each group (A: normal group; B: model group; C: Fuke Qianjin tablets group; D: low-dose MDE group; E: medium-dose MDE group; F: high-dose MDE group);

[0035] Figure 10 Pathological morphology of uterine tissues of rats in each group (HE×400) (A is the normal group; B is the model group; C is the positive group; D is the low-dose MDE group; E is the medium-dose MDE group; F is the high-dose MDE group). Specific implementation manners

[0036] The technical solution of the present invention will be further specifically described below through specific embodiments.

[0037] Example 1 Formula of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0038] 6 g of Melastoma dodecandrum Lour. extract, 11.2 g of poloxamer 407, 2.8 g of poloxamer 188, 12 mL of isopropyl myristate, 10 mL of Tween-80, 2 mL of glycerol.

[0039] Example 2 Formula of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0040] 8 g of Melastoma dodecandrum Lour. extract, 12.8 g of poloxamer 407, 4.4 g of poloxamer 188, 14 mL of isopropyl myristate, 14 mL of Tween-80, 3 mL of glycerol.

[0041] Example 3 Formula of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0042] 4 g of Melastoma dodecandrum Lour. extract, 9.6 g of poloxamer 407, 1.2 g of poloxamer 188, 6 mL of isopropyl myristate, 6 mL of Tween-80, 1 mL of glycerol.

[0043] Example 4 Formula of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0044] 7 g of Melastoma dodecandrum Lour. extract, 12 g of poloxamer 407, 3.6 g of poloxamer 188, 12 mL of isopropyl myristate, 12 mL of Tween-80, 2 mL of glycerol.

[0045] Example 5 Formula of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0046] 5 g of Melastoma dodecandrum Lour. extract, 10.4 g of poloxamer 407, 2 g of poloxamer 188, 10 mL of isopropyl myristate, 8 mL of Tween-80, 1 mL of glycerol.

[0047] The formulas of Examples 1-5 are used in the following preparation method

[0048] Example 6 Preparation method of Melastoma dodecandrum Lour. temperature-sensitive in-situ gel

[0049] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal material, add 14 times the amount of 95% ethanol, reflux extract twice, 1 hour each time, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, extract the suspension with petroleum ether (1:1) until colorless, take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0050] Weigh 11.2 g of poloxamer 407 and 2.8 g of poloxamer 188 into a beaker, add 100 mL of ultrapure water to swell, place in a -4°C refrigerator for 24 h until clear, add 2 mL of glycerol, and mix well to obtain Solution A; take 6 g of the Melastoma dodecandrum Lour. extract, dissolve it in 12 mL of isopropyl myristate in a 50°C water bath, add 10 mL of Tween-80, and mix well to obtain Solution B; add Solution A to Solution B with continuous stirring and stir for 10 min until uniform to obtain the product.

[0051] Preparation method of temperature-sensitive in-situ gel of Melastoma dodecandrum Lour. in Example 7

[0052] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal material, add 14 times the amount of 95% ethanol, reflux extract twice, 1 hour each time, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, extract the suspension with petroleum ether (1:1) until colorless, take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0053] Weigh 12.8 g of poloxamer 407 and 4.4 g of poloxamer 188 into a beaker, add 120 mL of ultrapure water to swell, place in a -4°C refrigerator for 28 h until clear, add 3 mL of glycerol, and mix well to obtain Solution A; take 8 g of the Melastoma dodecandrum Lour. extract, dissolve it in 14 mL of isopropyl myristate in a 60°C water bath, add 14 mL of Tween-80, and mix well to obtain Solution B; add Solution A to Solution B with continuous stirring and stir for 20 min until uniform to obtain the product.

[0054] Preparation method of temperature-sensitive in-situ gel of Melastoma dodecandrum Lour. in Example 8

[0055] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal material, add 14 times the amount of 95% ethanol, reflux extract twice, 1 hour each time, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, extract the suspension with petroleum ether (1:1) until colorless, take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0056] Weigh 9.6 g of poloxamer 407 and 1.2 g of poloxamer 188 into a beaker, add 80 mL of ultrapure water to swell, place in a -4°C refrigerator for 20 h until clear, add 1 mL of glycerol, and mix well to obtain Solution A; take 4 g of the Melastoma dodecandrum Lour. extract, dissolve it in 6 mL of isopropyl myristate in a 40°C water bath, add 6 mL of Tween-80, and mix well to obtain Solution B; add Solution A to Solution B with continuous stirring and stir for 5 min until uniform to obtain the product.

[0057] Preparation Method of Melastoma dodecandrum Lour. Temperature-Sensitive In-Situ Gel in Example 9

[0058] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal materials, add 14 times the amount of 95% ethanol, reflux extract twice, 1 hour each time, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, and extract the suspension with petroleum ether (1:1) until colorless. Take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0059] Weigh 12 g of poloxamer 407 and 3.6 g of poloxamer 188 into a beaker, add 110 mL of ultrapure water to swell, place it in a -4°C refrigerator for 26 h until it becomes clear, add 2 mL of glycerol, and mix evenly to obtain Solution A; take 7 g of the Melastoma dodecandrum Lour. extract, dissolve it in 12 mL of isopropyl myristate in a 55°C water bath, add 12 mL of Tween-80, and mix evenly to obtain Solution B; add Solution A to Solution B under continuous stirring and stir for 15 min until it is uniform, then it is obtained.

[0060] Preparation Method of Melastoma dodecandrum Lour. Temperature-Sensitive In-Situ Gel in Example 10

[0061] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal materials, add 14 times the amount of 95% ethanol, reflux extract twice, 1 hour each time, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, and extract the suspension with petroleum ether (1:1) until colorless. Take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0062] Weigh 10.4 g of poloxamer 407 and 2 g of poloxamer 188 into a beaker, add 90 mL of ultrapure water to swell, place it in a -4°C refrigerator for 24 h until it becomes clear, add 1 mL of glycerol, and mix evenly to obtain Solution A; take 5 g of the Melastoma dodecandrum Lour. extract, dissolve it in 10 mL of isopropyl myristate in a 45°C water bath, add 8 mL of Tween-80, and mix evenly to obtain Solution B; add Solution A to Solution B under continuous stirring and stir for 10 min until it is uniform, then it is obtained.

[0063] To verify the effectiveness of the present invention, the invention team conducted a series of tests, which are specifically as follows:

[0064] 1 Materials and Methods

[0065] 1.1 Materials

[0066] 1.1.1 Reagents and Medicinal Materials

[0067] Melastoma dodecandrum Lour. was collected in Huishui County, Guizhou Province, China in July 2024, identified by Professor Sun Qingwen, and the voucher specimens are preserved in Guizhou University of Traditional Chinese Medicine.

[0068] Poloxamer 407, 188 (P188, P407), bovine serum albumin (Beijing Solarbio Science & Technology Co., Ltd.). Isopropyl myristate (IPM), glycerol, urea (Shanghai Macklin Biochemical Co., Ltd.). Tween-80 (Shanghai Aladdin Biochemical Technology Co., Ltd.). KOH, Ca(OH)2 (Xilong Chemical Co., Ltd.), glucose (Sigma-Aldrich), lactic acid (Alfa Aesar Chemical Co., Ltd.). NaCl, acetic acid (Chengdu Jinshan Chemical Reagent Co., Ltd.). Fufang Qianjin Tablets (Zhuzhou Qianjin Pharmaceutical Co., Ltd.). Absorbable gelatin sponge (Jiangxi Xiangen Medical Technology Development Co., Ltd.). Progesterone injection (Zhejiang Xianju Pharmaceutical Co., Ltd.). TNF-α, IL-6 kits (Wuhan Elabscience Biotechnology Co., Ltd.).

[0069] 1.1.2 Instruments

[0070] Cryo-scanning electron microscope (Cryo-EM; Hitachi SU8010, Hitachi, Ltd.); Rotary rheometer (Waters DHR-2, TA Instruments, USA); Fourier transform infrared spectrometer (Thermo Fisher Scientific Nicolet iS20, Thermo Fisher Scientific); Differential scanning calorimeter (TA DISCOVERY 5500, TA Instruments, USA); Rotary microtome (Leica-2016, Leica Instruments GmbH, Germany); Automatic dehydrator (JT-12S, Wuhan Junjie Electronic Co., Ltd.); Tissue embedding machine (BMJ-A, Changzhou Suburban Zhongwei Electronic Instrument Factory); Microplate reader (SpectraMAX Plus384, Molecular Devices, USA), etc.

[0071] 1.1.3 Bacteria and animals

[0072] Escherichia coli (E. coli, ATCC25922) and Staphylococcus aureus (S. aureus, ATCC29213) were both purchased from Shanghai Luwei Technology Co., Ltd. SD female rats, SPF grade, weighing 200±20 g, were purchased from Changsha Tianqin Biotechnology Co., Ltd., and the experimental animal production license number was: SCXK(Xiang)2019-0014.

[0073] 1.2 Methods

[0074] 1.2.1 Preparation of Melastoma dodecandrum Lour. extract (MDE)

[0075] Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal materials, add 14 times the amount of 95% ethanol, reflux and extract twice, each time for 1 hour, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, and extract the suspension with petroleum ether (1:1) until colorless. Take the petroleum ether layer, recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

[0076] 1.2.2 Preparation of Melastoma dodecandrum Lour. in-situ gel (MD-TSIG)

[0077] Weigh appropriate amounts of P407 and P188 into a beaker, add ultrapure water to swell, place it in a -4°C refrigerator for 24 h until it becomes clear, add an appropriate amount of glycerol, and mix well to obtain Solution A. Dissolve MDE in IPM in a 50°C water bath, add Tween-80, and mix well to obtain Solution B (abbreviated as MdE / IPM). Add Solution A to Solution B with continuous stirring and stir evenly to obtain the product.

[0078] The gelation temperature is an important index for investigating temperature-sensitive gels. The literature method was slightly modified to measure the gelation temperature.

[13] . Take the MD-TSIG solution in a vial, place it in a 20°C water bath, with the liquid level in the vial 1 cm lower than the liquid level in the water bath. Heat it up, keep it for 1 min for every 1°C increase, then tilt the vial at 45°, observe the flow of the liquid, and the temperature at which the liquid no longer flows is the gelation temperature of the gel. Considering body temperature and the actual production environment, the gelation temperature of the temperature-sensitive gel should be lower than body temperature and higher than the production environment temperature. Therefore, the optimal gelation temperature of the temperature-sensitive gel is set at 31 - 34°C.

[14] . Each group of experiments was repeated three times, and the average value of the gelation temperatures of the three experiments was used as the index.

[0079] 1.2.3 Optimization of the MD-TSIG formulation by response surface methodology

[0080] Since MDE is poorly soluble in water, the solubility of MDE in solvents (olive oil, liquid paraffin, soybean oil, ethyl acetate, IPM) was investigated. After preliminary pre-experiments, the effects of the dosages of each component, namely P407 (9.6, 10.4, 11.2, 12.0, 12.8 g), P188 (1.2, 2.0, 2.8, 3.6, 4.4 g), IPM (6.0, 8.0, 10.0, 12.0, 14.0 mL), Tween-80 (6.0, 8.0, 10.0, 12.0, 14.0 mL), glycerol (1.0, 2.0, 3.0 mL), and the stirring time (5, 10, 15, 20 min) on the gelation temperature were further investigated. According to the results of the single-factor experiments, the dosages of P407 (A), P188 (B), and IPM (C) were selected as the investigation factors, and the gelation temperature was used as the response value. A Box-Behnken experimental design with 3 factors and 3 levels was carried out, with a total of 17 experiments, and each experiment was repeated 3 times. The factors and levels are shown in Table 1.

[0081] Table 1 Factor and level design

[0082]

[0083] 1.2.4 Quality characterization of Melastoma dodecandrum Lour. in-situ gel

[0084] Observe the appearance changes of MD-TSIG before and after coagulation against a white background. Observe the microstructure of MD-TSIG under Cryo-EM

[0085] 1.2.5 pH determination and centrifugal stability investigation of Melastoma dodecandrum Lour. in-situ gel

[0086] Take an appropriate amount of MD-TSIG and stir it in a thermostatic heating magnetic stirrer with a heating and magnetic stirring function at 20 r / min for 15 min under ice bath conditions to make the hydrogel solution uniform. After standing, insert the probe of the pH meter into the solution and read the data after the pH value stabilizes. Set up 3 parallel groups for each experiment and take the average value. Place the sample solution at 4, 25, and 37 °C for 20 min respectively, centrifuge at 3000 r / min for 30 min, and observe whether there are unstable phenomena such as stratification

[15] 。

[0087] 1.2.6 Rheological determination of Melastoma dodecandrum Lour. in-situ gel

[0088] Use a rotary rheometer to perform rheological analysis on MD-TSIG. Under the oscillatory scanning mode, set the strain (γ*) to be 0.01 g to 100 g, and test the curves of the storage modulus (G′) and loss modulus (G″) of the gel varying with γ* at 4, 25, and 37 °C. Under the oscillatory mode, at a temperature of 37 °C, a strain amplitude of 1 g, and a scanning frequency (f) of 0.1 to 10 Hz, perform a frequency scan, and record the curves of the rheological parameters of the gel varying with f at 4, 25, and 37 °C respectively. Set the temperature change to be 4 to 50 °C and the heating rate to be 2 °C / min, test the curves of G′ and G″ varying with temperature, and determine the phase transition temperature. The temperature at the intersection of the curves is regarded as the gelation temperature

[16] 。

[0089] 1.2.7 Fourier transform infrared spectroscopy determination of Melastoma dodecandrum Lour. in-situ gel

[0090] Lyophilize MD-TSIG to obtain a green sponge-like sample, and use a Fourier transform infrared spectrometer for testing. Set the wavenumber range to be 4000 - 400 cm -1[17-18] 。

[0091] 1.2.8 Thermal performance analysis of Melastoma dodecandrum Lour. in-situ gel

[0092] Thermodynamic tests were carried out using a differential scanning calorimeter to study the interaction between water and MD-TSIG. The sample was accurately weighed and loaded into a sealed aluminum pan, and scanned at a standard heating rate of 10 °C / min from 30 to 800 °C

[19] .

[0093] 1.2.9 Influence of dilution with vaginal simulant

[0094] According to the literature

[20] described therein, the simulated vaginal fluid was prepared. Weigh 1.4 g of KOH, 0.222 g of Ca(OH)2, 0.018 g of bovine serum albumin, 3.510 g of sodium chloride, 5.000 g of glucose, 0.160 mL of glycerol, 2.000 g of lactic acid, 0.400 g of urea and 1.000 g of acetic acid respectively. Add 1 L of water and stir to dissolve. Adjust the pH value to 4.2 - 4.5 with hydrochloric acid. Mix MD-TSIG with the simulated vaginal fluid at a ratio of 40:7.5

[21] , and the diluted MD-TSIG was obtained. Measure the complex viscosity (η*) of MD-TSIG when heated from 4 °C to 50 °C before and after dilution with the simulated vaginal fluid

[22] .

[0095] 1.2.10 In vitro antibacterial study of Melastoma dodecandrum Lour. in-situ gel

[0096] Using the streak plate method, E. coli and S. aureus were inoculated on the agar medium plate respectively, and incubated overnight in a constant temperature incubator at 37 °C. Single colonies were picked and inoculated into a centrifuge tube containing agar medium, and cultured at 37 °C and 180 r / min for 24 h. The bacterial culture solution was diluted to 1×10 5 CFU / mL and stored at 4 °C for later use. Take 0.5 and 1 g of MD-TSIG in centrifuge tubes, then add the bacterial suspension. The mixture was shaken and cultured at 37 °C for 24 h. After the culture, take 50 μL of the bacterial suspension and spread it on the agar plate, and culture it in the incubator for 16 h. Finally, record the growth of E. coli and S. aureus

[23] .

[0097] 1.2.11 Study on anti-inflammatory effect in vivo

[0098] The petroleum ether extract of Melastoma dodecandrum Lour. has good in vitro anti-inflammatory activity, so a rat model of cervicitis was used to study the anti-inflammatory pharmacodynamics in vivo

[0099] (1) Establish a rat model of cervicitis

[24]

[0100] Take Escherichia coli and Staphylococcus aureus in the logarithmic growth phase, and adjust the concentration to 1×10 9Individuals / mL, mix evenly at a ratio of 1:1 for standby. The rats were fed adaptively for 7 days, and each rat was injected subcutaneously with 10 mg / kg progesterone. After 1 week, a 0.125 mL absorbable gelatin sponge (0.5 cm * 0.5 cm) was immersed in a mixed bacterial solution of Staphylococcus aureus and pathogenic Escherichia coli, and the mixed bacterial solution was fully absorbed. Except for the control group rats, the upper reproductive tracts of all rats were inserted with the gelatin sponge containing microorganisms, and then the rats were forced to lie on the ground for 3 minutes. This operation was performed four times at intervals of 2 days for infection, and the normal group rats were not treated.

[0101] (2) Drug administration and sample collection

[0102] The female SD rats were randomly divided into groups, including a normal group, a model group, a Fuke Qianjin Pian group (0.68 g / kg), high, medium, and low dose groups of MDE (14 g / kg, 7 g / kg, 3.5 g / kg). The drugs in each group were ground with 0.5 g of sodium carboxymethylcellulose into a suspension. The normal group and the model group were given an equal volume of 0.5 g of sodium carboxymethylcellulose. After the first infection, gavage administration was started, and the administration volume was 10 mL / kg, and the administration was continued for 10 days. After the last administration, the rats were fasted but not water-deprived, and anesthetized by intraperitoneal injection of 20 g of ethyl carbamate (5 mL / kg). Blood was taken from the abdominal aorta, allowed to stand, and centrifuged at 3500 rpm for 10 minutes at 4 °C. The serum was separated and aliquoted into sealed test tubes for low-temperature storage for standby.

[0103] (3) Observation of uterine changes in rats and HE staining to observe the pathological changes of uterine tissue

[0104] After blood collection, the rats were sacrificed by decapitation. The uterine tissue of the rats was taken, and part of the uterine tissue was fixed with 4 g of paraformaldehyde. After dehydration, trimming, embedding, sectioning, staining, and sealing, the pathological changes of the uterine tissue of rats in each group were observed under an optical microscope.

[0105] 2 Results and discussion

[0106] 2.1 Single factors affecting the gelation temperature

[0107] The solubility of MDE in olive oil, liquid paraffin, soybean oil, ethyl acetate, and IPM from high to low is: ethyl acetate > IPM > soybean oil > liquid paraffin > olive oil. Although the solubility of ethyl acetate is the highest, IPM was selected as the best solvent because of its non-volatility and pharmaceutical-grade compatibility. From Figure 1 It can be seen that the dosages of P407, P188, and IPM have a significant effect on the gelation temperature. Therefore, in the subsequent experiments, P407 (9.6 g, 10.4 g, 11.2 g), P188 (2.0 g, 2.8 g, 3.6 g), and IPM (8.0 g, 10.0 g, 12.0 g) were determined.

[0108] 2.2 Model establishment and significance analysis

[0109] The Box-Behnken response surface model was applied to simultaneously optimize the levels of each variable to achieve the best system performance and statistical prediction was carried out. The experimental arrangements and results are shown in Table 2.

[0110] Table 2 Box-Behnken response surface experimental design and results

[0111]

[0112] Using Design Expert 13 software, with the gelling temperature as the response value, the experimental data were subjected to quadratic multiple regression fitting analysis, and the following quadratic multiple regression equation was obtained:

[0113] R = 32.20 - 6.37A - 7.25B + 2.38C - 0.75AB + 2.00AC - 0.25BC + 3.5A 2 + 0.75B 2 - 0.50C 2 , the P value of the model used in the regression equation is < 0.01, and the P value of the lack-of-fit term is 0.7022 > 0.05. The correlation coefficient R of the model 2 = 0.9662, and the adjusted determination coefficient R 2 Adj = 0.9228, indicating that the model has good credibility, there are no lack-of-fit factors and it can fit well with the actual values, and the model can explain the changes in the response value.

[0114] This model was used to predict and analyze the preparation process of MD-TSIG, and the results are shown in Table 3. Among them, factors A, B, and C have extremely significant effects on the linear effect of the gelling temperature (P < 0.01). By comparing the magnitudes of the F values, it can be seen that the order of the effects of each factor on the gelling temperature is B (P188) > A (P407) > C (IPM). The response surface diagram can intuitively reflect the influence of the interaction on the predicted value. When the response surface curve is steeper, the influence of these two interaction factors on the response value is more significant. On the contrary, when the response surface curve is not very steep, it is considered that the influence is not significant. See specifically Figure 2 , and this figure is consistent with the results of the variance analysis.

[0115] Considering that the dilution of vaginal fluid causes an increase in the gelation temperature, the gelation temperature is selected to be between 33 °C and 34 °C. The target gelation temperature of the prescription is set at 33 °C. From the fitting equation, the optimal prescription of Melastoma dodecandrum Lour. in-situ gel MD-TSIG can be obtained, that is, 11.2 g of P407, 2.8 g of P188, and 12 mL of IPM. Five batches of MD-TSIG were prepared under the optimal preparation process conditions, and their gelation temperatures were measured to be 33.4 °C, indicating that the mathematical model established by the Box-Behnken response surface method has good predictability, and the optimized MD-TSIG preparation process conditions are accurate and reliable.

[0116] Table 3 Analysis of variance of the regression model

[0117]

[0118] 2.3 Investigation of appearance and microscopic morphology

[0119] The state changes of MD-TSIG at room temperature and 37 °C are shown in Figure 3 a. Under normal temperature conditions, the preferred preparation sample is in the form of a dark green solution, showing fluidity in an inverted vial, and the sample pH is 5.29 ± 0.03, meeting the vaginal administration preparation standard (4.5 - 5.5).

[25] . When the external temperature rises to the gelation temperature, the preparation shows a semi-solid gel form and does not flow with the inversion of the vial, indicating that the preparation has completed the phase transition process from the solution state to the gel state at this time. The microscopic structure of freeze-dried MD-TSIG was observed under Cryo-SEM (see Figure 3 b, c), and it was found that the drug-loaded gel is a three-dimensional cross-linked polymer network structure, and pores of different sizes are formed during polymer cross-linking.

[0120] 2.4 Rheological measurement

[0121] The results of the linear viscoelastic region scan are shown in Figure 4 a. At 4 °C, when γ* starts to increase, the elastic storage modulus G′ and the viscous loss modulus G″ remain relatively constant. G″ > G′ indicates that the viscosity of the gel dominates at this time; at 25 °C and 37 °C, when γ* starts to increase, the elastic storage modulus G′ and the viscous loss modulus G″ remain relatively constant. G′ > G″ indicates that the elasticity of the gel dominates at this time. As the strain continues to increase, both G′ and G″ show a downward trend, indicating that the gel structure begins to be damaged. G′ rapidly decreases and is lower than G″, indicating that the cross-linked structure inside the hydrogel is damaged under high strain, and the gel state changes to the solution state. Therefore, all dynamic oscillation experiments need to control γ* within 1 g. The results of the frequency scan are shown in Figure 4b. At 4 °C, G″ > G′, showing fluid characteristics; at 25 °C and 37 °C, G′ > G″, showing elastic characteristics, no frequency dependence was observed, and MD-TSIG showed viscoelastic solid properties and had a stable three-dimensional network structure. The temperature scan results are shown in Figure 4 c. When the temperature was 21.19 °C, G′ = G″, which was the phase transition temperature of MD-TSIG. The gelling temperature measured by the inverted test tube method was the temperature when the micelles in the system aggregated to form a semi-solid gel, rather than the starting temperature of gel formation. Therefore, the gelling temperature was higher than the phase transition temperature.

[14] .

[0122] 2.5 Fourier Transform Infrared Spectroscopy

[0123] In the infrared spectrum of poloxamer, the stretching vibration peak of associated -OH was at 3481 cm -1 , the stretching vibration peak of -CH was at 2888 cm -1 , and the stretching vibration peak of -CO- was at 1108 cm -1 . Compared with poloxamer, the stretching vibration peak of -OH in Melastoma dodecandrum Lour. in-situ gel shifted to 3376 cm -1 . The displacement of these absorption peaks might be the result of intermolecular hydrogen bond interaction. In the infrared spectrum of MD-TSIG, most of the characteristic absorption peaks of poloxamer and MdE / IPM were retained, and no obvious disappearance of each characteristic peak was observed. Only the positions of some peaks changed, indicating good compatibility between poloxamer and MdE / IPM. They interacted mainly through intermolecular forces (such as hydrogen bonds), and no obvious chemical reaction occurred to cause the destruction of functional groups or the formation of new functional groups. The results are shown in Figure 5 .

[0124] 2.6 Thermal Property Analysis

[0125] The thermogravimetric analysis / differential thermogravimetric analysis (TG / DTG) curves of MD-TSIG are shown in Figure 6 . It can be seen from the figure that MD-TSIG had two obvious weight losses between 30 and 800 °C. The first stage occurred at about 116 °C, with a weight loss of about 64.85 g. This weight loss was caused by the evaporation of residual water in the polymer and the decomposition of the drug. Because MD-TSIG had hydrophilic amide groups, it could store water internally. When the temperature increased, the water separated from the polymer and was completely evaporated in an environment above 100 °C. The second weight loss occurred at about 413 - 434 °C, with a weight loss of about 22.12 g. In this stage, partial decomposition of MD-TSIG occurred. It can be seen from the TG / DTG curves that when the temperature reached 116 °C, the peak of the DTG curve also reached the maximum value, indicating that the decomposition rate of MD-TSIG reached the maximum. When the temperature reached about 450 °C, MD-TSIG was basically decomposed.

[0126] 2.7 Influence of dilution with vaginal simulation fluid

[0127] Viscosity is a key rheological characteristic of temperature-sensitive gels. At low temperatures, the temperature-sensitive gel has low viscosity and is in a flowing liquid state, which can quickly spread and cover the surface of vaginal folds. After phase transition, its viscosity increases sharply and the gel strength becomes larger, which can stay locally in the vagina for a long time. The gelation temperature of MD-TSIG after dilution with simulated vaginal fluid is about 35.6 °C, and the appearance is shown in Figure 7 b. After dilution with vaginal simulation fluid, the viscosity decreases slightly compared with that before dilution, indicating a decrease in gel strength, but still showing good gelation strength. The results are shown in Figure 7 a. MD-TSIG did not show delamination after centrifugation after being placed at 4, 25, and 37 °C for a period of time. The results are shown in Figure 7 c.

[0128] 2.8 In vitro antibacterial study

[0129] The antibacterial experiment results show that MD-TSIG has antibacterial properties against both S. aureus and E. coli. With the increase of the concentration of MD-TSIG, its antibacterial effect becomes more obvious. The results are shown in Figure 8 .

[0130] 2.9 Observation of histopathological changes in rat uterus by uterine tissue and HE staining

[0131] After taking blood, the rats were sacrificed by cervical dislocation. The uterine tissues of the rats were taken, and part of the uterine tissues were selected and fixed with 4 g paraformaldehyde. After dehydration, trimming, embedding, sectioning, staining, and mounting, the pathological changes of the uterine tissues of each group of rats were observed under an optical microscope.

[0132] As Figure 9 , 10 shown, the sizes, shapes, colors of the uteri of the rats in the normal group were normal, symmetrical on the left and right, elastic, without congestion and edema, and occasional inflammatory cells were scattered in the stroma and muscular layer; the uterus of the rats in the model group was swollen in shape, congested and thickened, asymmetrical in shape, and different degrees of inflammatory cell infiltration appeared in the stroma, mainly lymphocytes, neutrophils and eosinophils, indicating successful modeling. Compared with the normal group, the pathological changes of the uterine tissues of the rats in the model group were more severe; the uteri of the rats in each drug administration group had varying degrees of swelling and congestion, and were relatively asymmetrical on the left and right, and all had varying degrees of chronic inflammatory infiltration; compared with the model group, the degree of uterine swelling and congestion in each group of rats was lighter, and the degree of inflammatory cell infiltration in the uterine tissues of the low-dose MDE group and the Fufang Qianjinpian group was relatively lighter, and the degree of inflammatory cell infiltration in the uterine tissues of the medium- and high-dose MDE groups was relatively the lightest.

[0133] In summary, the final formulation of the Melastoma dodecandrum Lour. in-situ gel is as follows: Melastoma dodecandrum Lour. extract 4.18%, P188 1.95%, P407 7.80%, IPM 6.96%, Tween-80 7.66%, glycerol 1.81%, ultrapure water 69.64%, that is, Melastoma dodecandrum Lour. extract 6 g, P407 11.2 g, P188 2.8 g, IPM 12 mL, Tween-80 10 mL, glycerol 2 mL, ultrapure water 100 mL.

[0134] Although the present invention has been described in detail above with general descriptions, specific embodiments and experiments, based on the present invention, some modifications or improvements can be made thereto, which are obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection of the present invention.

[0135] References

[0136] [1] Ming Huiyi, Ma Xiuping*, Yang Jing, et al. Analysis of the current research status of the ethnic medicine Melastoma dodecandrum Lour. [J]. Chinese Journal of Experimental Traditional Medical Formulae. 2018, 24(11): 228-234.

[0137] [2] Wang JF, Jia ZY, Zhang ZH, et al. Analysis of Chemical Constituents of Melastoma dodecandrum Lour. by UPLC-ESI-Q-Exactive Focus-MS / MS [J]. Molecules, 2017, 22(3): 476.

[0138] [3] Zhang ZQ, Xu Y, Liang JM, et al. Polyphenolics and triterpenoids from the whole herbs of Melastoma dodecandrum Lour. and their anti-inflammatory activity [J]. Fitoterapia, 2025, 180: 106337.

[0139] [4] Huang G, Ge YL, Gui ZH, et al. Toxicity of Lour. and its effects on lipopolysaccharide-induced inflammation and oxidative stress[J]. Experimental and therapeutic medicine, 2021, 22(2): 807.

[0140] [5] Qian S, Wang ZH, Ma XP*, et al. Study on the Active Components and Inhibiting Effect of Melastoma dodecandrum Lour. on Human Cervical Cancer Cells Based on Spectrum–Effect Relationship Analysis[J]. Natural Product Communications, 2024, 19(1): 1-12.

[0141] [6] Fang CW, Ma XP*, Liu J, et al. Study on the anti-inflammatory spectrum-effect relationship of chloroform extract of Melastoma dodecandrum Lour. based on multivariate statistical analysis[J]. Journal of Chinese Medicinal Materials, 2024, 52(6): 1454-1460.

[0142] [7] Wang WX, Qin XY, Ma XP*, et al. Study on the spectrum-effect relationship of the hemostatic effect of Melastoma dodecandrum Lour. and preliminary exploration of its in vitro coagulation activity[J]. Journal of Chinese Medicinal Materials, 2022, 45(1): 130-136.

[0143] [8] Luo SX, Ma XP*, Liu J, et al. Basic research on anti-inflammatory active substances of petroleum ether extract of Melastoma dodecandrum Lour. based on spectrum-effect relationship[J]. China Pharmacy, 2024, 35(23): 2877-2882.

[0144] [9] Sun X, Ning HX, Hou WB, et al. Research progress on traditional Chinese medicine in-situ gels for vaginal use[J]. Chinese Traditional and Herbal Drugs, 2022, 53(11): 3564-3572.

[0145]

[10] Soliman GM, Fetih G, Abbas AM. Thermosensitive bioadhesive gels for the vaginal delivery of sildenafil citrate: in vitro characterization and clinical evaluation in women using clomiphene citrate for induction of ovulation[J]. Drug development and industrial pharmacy, 2016, 43(3): 399-408.

[0146]

[11] Wang XQ, Liu YL, Lin LF, et al. Rheological and in vitro release studies of temperature-sensitive in-situ gels of Yihuang Decoction for vaginal use and its ordinary gels[J]. China Journal of Chinese Materia Medica, 2020, 45(03): 539-547.

[0147]

[12] Wan J, Li XX, Chen C, et al. Research progress on environmentally responsive in-situ gel matrices[J]. West China Journal of Pharmaceutical Sciences, 2024, 39(06): 727-732.

[0148]

[13] Zhang SJ, Zhang YT, Wang Z, et al. Temperature-sensitive gel-loaded composite nanomedicines for the treatment of cervical cancer by vaginal delivery. International journal of pharmaceutics, 2020, 586: 119616.

[0149]

[14] Lin JY, Chen YJ, Xu BH, et al. Preparation and in vitro drug release of temperature-sensitive progesterone in-situ gel[J]. Journal of China Pharmaceutical University, 2014, 45(03): 325-330.

[0150]

[15] Zhang DD, Wu YL, Wei HH, et al. Preparation and property investigation of temperature-sensitive gel of Schizonepeta tenuifolia Briq. oil-sophoridine vaginal liposomes[J]. Chinese Traditional and Herbal Drugs, 2014, 45(07): 929-934.

[0151]

[16] Bai Jiaojiao, Ding Yan, Sabahaiti Ablimit, et al. Preparation and Characterization of Gallic Acid pH / Temperature-Sensitive Gels [J]. Chinese Journal of New Drugs, 2024, 33(22): 2379-2389.

[0152]

[17] Zhang Y, Dou X, Zhang L, et al. Facile fabrication of a biocompatible composite gel with sustained release of aspirin for bone regeneration [J]. Bioactive Materials, 2021, 11: 130-139.

[0153]

[18] Li R, Li R, Lin Z, et al. Injectable and -Formable Thiolated Chitosan-Coated Liposomal Hydrogels as Curcumin Carriers for Prevention of Breast Cancer Recurrence [J]. ACS applied materials & interfaces, 2020, 12(15): 17936-17948.

[0154]

[19] Shriky B, Kelly A, Isreb M, et al. Pluronic F127 thermosensitive injectable smart hydrogels for controlled drug delivery system development [J]. Journal of colloid and interface science, 2019, 565: 119-130.

[0155]

[20] Abd Ellah Noura H,Abdel-Aleem Jelan A,Abdo Mariana N,etal.Efficacy of ketoconazole gel-flakes in treatment of vaginal candidiasis:Formulation,in vitro and clinical evaluation[J].International journal ofpharmaceutics,2019,567:118472.

[0156]

[21] Chang JY,Oh YK,Choi HG,et al.Rheological evaluation ofthermosensitive and mucoadhesive vaginal gels in physiological conditions[J].IntJ Pharm,2002,241(1):155.

[0157]

[22] Yuan Ping,Chen Ying,Liu Lina.Study on rheological properties and sustained-release retention of octoxynol vaginal thermosensitive gel[J].China Pharmacist,2017,20(06):982-985.

[0158]

[23] Xue Haiyan,Zhou Yan,He Baoyuan,et al.Preparation and antibacterial properties of polydopamine silver nanoparticles enhanced hydrogels[J / OL].Journal of Shaanxi University of Science & Technology,1-9[2025-03-12].https: / / doi.org / 10.19481 / j.cnki.issn2096-398x.20241023.002.

[0159]

[24] Guo Shanshan,Gao Yingjie,Bao Lei,et al.In vivo and in vitro pharmacodynamic evaluation of Zijin Huadu suppository in the treatment of cervicitis[J].Chinese Journal of Experimental Traditional Medical Formulae,2016,22(5):115-122.

[0160]

[25] Hou Li,Liu Chang,Xin Ling,et al.Preparation and evaluation of antibacterial peptide Maximin11 vaginal gel[J].Chinese Journal of New Drugs,2019,28(01):81-86.

Claims

1. A Melastoma dodecandrum Lour. temperature-sensitive in-situ gel, characterized in that, The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel is composed of 4 - 8 g of Melastoma dodecandrum Lour. extract, 9.6 - 12.8 g of poloxamer 407, 1.2 - 4.4 g of poloxamer 188, 6 - 14 mL of isopropyl myristate, 6 - 14 mL of Tween-80, and 1 - 3 mL of glycerol.

2. The in-situ gel of Melastoma dodecandrum Lour. with temperature sensitivity according to claim 1, wherein, The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel is composed of 5 - 7 g of Melastoma dodecandrum Lour. extract, 10.4 - 12.0 g of poloxamer 407, 2.0 - 3.6 g of poloxamer 188, 10 - 14 mL of isopropyl myristate, 8 - 12 mL of Tween-80, and 2 - 3 mL of glycerol.

3. The in-situ gel of Melastoma dodecandrum Lour. with temperature sensitivity according to claim 2, wherein, The Melastoma dodecandrum Lour. temperature-sensitive in-situ gel is composed of 6 g of Melastoma dodecandrum Lour. extract, 11.2 g of poloxamer 407, 2.8 g of poloxamer 188, 12 mL of isopropyl myristate, 10 mL of Tween-80, and 2 mL of glycerol.

4. The thermosensitive in-situ gel of Melastoma dodecandrum Lour. according to claims 1-3, characterized in that The preparation method of the Melastoma dodecandrum Lour. extract is as follows: Take 0.2 kg of the crude powder of Melastoma dodecandrum Lour. medicinal materials, add 14 times the amount of 95% ethanol, reflux and extract twice, each time for 1 hour, filter, combine the filtrates, concentrate to a relative density of 1.02, suspend the extract in water, extract the suspension with petroleum ether at a ratio of 1:1 until colorless, take the petroleum ether layer to recover the solvent, and concentrate to dryness in a water bath to obtain the Melastoma dodecandrum Lour. extract.

5. A preparation method of the Melastoma dodecandrum Lour. temperature-sensitive in-situ gel according to any one of claims 1-3, characterized in that, The preparation method is as follows: Weigh 9.6 - 12.8 g of poloxamer 407 and 1.2 - 4.4 g of poloxamer 188 into a beaker, add 80 - 120 mL of ultrapure water to swell, place it in a -4°C refrigerator for 20 - 28 h until it becomes clear, add 1 - 3 mL of glycerol, and mix well to obtain Solution A; Take 4 - 8 g of Melastoma dodecandrum Lour. extract, dissolve it with 6 - 14 mL of isopropyl myristate in a water bath at 40 - 60°C, add 6 - 14 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 5 - 20 min until it is uniform, then it is obtained.

6. The preparation method of a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel according to claim 5, characterized in that, The preparation method is as follows: Weigh 10.4 - 12.0 g of poloxamer 407 and 2.0 - 3.6 g of poloxamer 188 into a beaker, add 90 - 110 mL of ultrapure water to swell, place it in a -4°C refrigerator for 22 - 26 h until it becomes clear, add 2 - 3 mL of glycerol, and mix well to obtain Solution A; Take 5 - 7 g of Melastoma dodecandrum Lour. extract, dissolve it with 10 - 14 mL of isopropyl myristate in a water bath at 45 - 55°C, add 8 - 12 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 10 - 15 min until it is uniform, then it is obtained.

7. The preparation method of a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel according to claim 6, characterized in that, The preparation method is as follows: Weigh 11.2 g of poloxamer 407 and 2.8 g of poloxamer 188 into a beaker, add 100 mL of ultrapure water to swell, place it in a -4°C refrigerator for 24 h until it becomes clear, add 2 mL of glycerol, and mix well to obtain Solution A; Take 6 g of Melastoma dodecandrum Lour. extract, dissolve it with 12 mL of isopropyl myristate in a water bath at 50°C, add 10 mL of Tween-80, and mix well to obtain Solution B; Add Solution A to Solution B while stirring continuously, and stir for 10 min until it is uniform, then it is obtained.

8. Use of a Melastoma dodecandrum Lour. temperature-sensitive in-situ gel according to any one of claims 1 - 7 in the preparation of a drug for inhibiting and anti-inflammatory of gynecological inflammation.

Images