Coptis chinensis detoxifying rinse, its preparation method and application

Abstract

This invention provides the application of Coptis chinensis detoxifying rinsing agent in the preparation of a medicament for treating or preventing infected wounds of the skin and / or mucous membranes. The Coptis chinensis detoxifying rinsing agent provided by this invention has significant antibacterial, anti-inflammatory, and wound-healing promoting effects, and is suitable for rinsing treatment of chronic infected wounds such as pressure injuries (PIs). The rinsing agent has a stable preparation process, controllable quality, and safe use, providing an effective traditional Chinese medicine external antibacterial rinsing solution for clinical application.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, specifically to a Coptis chinensis detoxification rinse, its preparation method, and its application. Background Technology

[0002] Pressure injuries (PIs), diabetic foot ulcers, and other chronic, refractory wounds are common and challenging clinical problems. These wounds are often accompanied by bacterial infection and persistent abnormal inflammatory responses, leading to stunted healing and even serious complications such as sepsis, increasing the risk of death for patients. Debridement is a fundamental and crucial step in managing these wounds, and irrigation is an important means of effective debridement. Irrigating the wound with sufficient amounts of irrigating fluid can physically remove necrotic tissue, purulent secretions, and bacteria, creating conditions conducive to wound healing.

[0003] Currently, the solutions used clinically for wound irrigation mainly include physiological saline, povidone-iodine, hydrogen peroxide, and chlorhexidine gluconate. While physiological saline is safe, it lacks active antibacterial activity; chemical disinfectants such as povidone-iodine and hydrogen peroxide, although antibacterial, may have cytotoxic effects on newly formed granulation tissue, inhibit wound healing, and long-term use may lead to bacterial resistance.

[0004] Traditional Chinese medicine (TCM) has unique advantages in anti-infection, anti-inflammatory, and tissue repair promotion, including multiple targets and low susceptibility to drug resistance. Huanglian Jiedu Decoction, from the "Elbow-Side Emergency Formulas," is a classic formula for clearing heat and detoxifying, composed of Coptis chinensis, Scutellaria baicalensis, Phellodendron chinense, and Gardenia jasminoides. Traditionally, it is administered orally. However, oral administration has inherent limitations in treating infected wounds. A study by Qin Kunming, Guo Hui, Xu Zisheng, et al. "Current Status of Research on Chemical Composition and Pharmacokinetics of Huanglian Jiedu Decoction [J]. Anti-infective Pharmacy, 2011, 8(01):3-7" indicates that after oral administration of Huanglian Jiedu Decoction, the active ingredients need to be absorbed through the gastrointestinal tract and metabolized by the liver before entering the bloodstream and reaching various tissues and organs through distribution. This means the active ingredients are largely metabolized or diluted before reaching the skin wound, making it difficult to achieve direct, high-concentration antibacterial and anti-inflammatory effects at the wound site, thus failing to meet the treatment needs of infected wounds.

[0005] To address this issue, some studies have attempted to use Huanglian Jiedu Decoction for external treatment. For example, the study "Duan Lianfeng, Wang Maosheng, Tao Yongling, et al. Observation on the efficacy of Huanglian Jiedu Decoction combined with moist burn ointment in the treatment of perianal infection after chemotherapy for acute leukemia [J]. Chinese Journal of Emergency Medicine, 2008, (09): 1223+1258" reported that the treatment of perianal infection using Huanglian Jiedu Decoction decoction for fumigation combined with moist burn ointment achieved good results. However, the decoction needs to be heated and used for local fumigation or sitz baths, which is cumbersome and difficult to standardize. More importantly, it still uses traditional decoctions and does not solve the inherent problems of the decoction itself.

[0006] Using traditional decoctions directly as rinsing solutions presents the following technical obstacles: (1) the decoctions contain a large number of impurities and tannins, which may cause irritation or allergic reactions when applied directly to open wounds; (2) the decoctions are not easy to preserve and are prone to microbial growth, failing to meet the sterility requirements of clinical rinsing solutions; (3) each use requires temporary decoction preparation, which is inconvenient for clinical application. Therefore, developing a stable, quality-controllable, convenient, safe, and effective external rinsing preparation of Huanglian Jiedu Decoction and applying it to the rinsing treatment of infected wounds is of great significance for enriching clinical treatment methods and meeting the urgent clinical needs of infected wounds. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a traditional Chinese medicine preparation based on Huanglian Jiedu Decoction, suitable for rinsing infected wounds of the skin and / or mucous membranes. This rinsing agent utilizes modern preparation processes to extract and purify the active ingredients, exhibiting stable properties and ease of storage and use.

[0008] The objective of this invention is achieved through the following technical solution: On one hand, the present invention provides the use of Coptis chinensis detoxifying rinse in the preparation of a medicament for treating or preventing infected wounds of the skin and / or mucous membranes; Furthermore, infected skin wounds include pressure injuries, diabetic foot ulcers, infected burn wounds, and / or infected surgical wounds.

[0009] On the other hand, the present invention provides a method for preparing a Coptis chinensis detoxifying rinse, comprising the following steps: S1: Weigh out the medicinal slices according to the mass ratio of Coptis chinensis: Phellodendron chinense: Scutellaria baicalensis: Gardenia jasminoides in 3:2:2:3, that is, 7.5g of Coptis chinensis, 5.0g of Phellodendron chinense, 5.0g of Scutellaria baicalensis, and 7.5g of Gardenia jasminoides to obtain mixed medicinal materials; S2: Add water to the mixed medicinal materials and decoct to extract, filter for the first time, collect the water extract, and obtain the filtrate; S3: Concentrate the filtrate under reduced pressure to obtain a concentrated solution; S4: The concentrate is subjected to alcohol precipitation, filtered a second time, and the supernatant is collected; S5: Recover the ethanol from the supernatant, adjust the pH, add sorbic acid, and add water to make up to 100 ml; S6: After the liquid has been brought to a certain volume, it is sterilized at high temperature and then dispensed to obtain the Coptis chinensis detoxification rinse solution. Furthermore, in step S2, the herbs are soaked in water for 2 hours before decoction, and then decocted 3 times for 1 hour each time, with the amount of water added being 14 times the total weight of the mixed herbs. Furthermore, in step S3, the concentration is carried out under reduced pressure and the temperature is controlled at 80°C. Furthermore, in step S4, 95% ethanol is added to the concentrate to make the final alcohol content of the system reach 80%, and alcohol precipitation is carried out under refrigeration for 12-24 hours. Furthermore, in step S5, the amount of sorbic acid added is 0.1%-0.3% of the final drug solution mass.

[0010] In another aspect, the present invention provides a Coptis chinensis detoxification rinse, wherein the Coptis chinensis detoxification rinse includes geniposide, baicalin, berberine hydrochloride and berberine hydrochloride as active ingredients; Furthermore, the Coptis chinensis detoxifying rinse is used to inhibit pro-inflammatory factors. IL-1β , IL-6 and TNF-α The expression of key proteins in the TLR2 / MyD88 / NF-κB inflammatory pathway was downregulated to reduce bacterial load in the wound and promote wound healing.

[0011] The beneficial effects of this invention are: This invention utilizes modern Chinese medicine preparation technology to develop the classic oral formula Huanglian Jiedu Decoction into a sterile external preparation that can be directly used for wound irrigation. This solves the problems of inconvenience, difficulty in preservation, and high impurity content of traditional decoctions for external use, and achieves convenience and standardization in clinical use.

[0012] This invention has been verified through animal experiments, confirming that the prepared irrigating agent can effectively reduce the bacterial load on wounds and inhibit pro-inflammatory factors ( ). IL-1β , IL-6 , TNF-α It expresses and downregulates key proteins in the TLR2 / MyD88 / NF-κB inflammatory pathway, thereby exerting multiple effects such as antibacterial, anti-inflammatory and wound healing promotion.

[0013] This invention demonstrates that the formulation has no significant irritation through irritation tests and process purification methods, as shown in rabbit skin and eye irritation tests. At the same time, impurities and tannins are removed through alcohol precipitation purification, and the pH is adjusted to be close to the physiological range of the skin, thereby significantly improving the safety of topical application.

[0014] This invention establishes a multi-index HPLC method for component content determination, using geniposide, baicalin, berberine hydrochloride, and berberine hydrochloride as quality control indicators. This achieves stable control over intermediates and finished products, ensuring process reproducibility and consistency in product efficacy. Furthermore, this invention provides a wound irrigation agent with traditional Chinese medicine characteristics, offering a new option for the clinical treatment of chronic infected wounds, particularly suitable for patients resistant to antibiotics or intolerant to chemical disinfectants, thus aligning with the modern concept of "wound bed preparation" in wound management. Attached Figure Description

[0015] Figure 1This is a graph showing the trend of wound healing rate in each group of rats in Example 3; Figure 2 This is a graph showing the trend of bacterial concentration changes in the wounds of rats in each group in Example 3; Figure 3 This is a comparison of the expression levels of TLR2, MyD88, and NF-κBp65 proteins in the wound tissues of rats in each group in Example 3; Figure 4 Figure 4 shows the results of skin irritation in rabbits in Example 4 (a. saline group; b. Coptis chinensis detoxification rinse group). Figure 5 The results of acute eye stimulation in rabbits in Example 4 are shown in the figure (a. saline group; b. Coptis chinensis detoxification rinse group). Detailed Implementation

[0016] The technical solutions in some embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments provided in this application, all other embodiments obtained by those skilled in the art are within the scope of protection of this application.

[0017] Example 1: Determination of Key Process Parameters The alcohol precipitation process was optimized using an orthogonal experimental design. Alcohol precipitation concentration and time were considered as factors, and the comprehensive score of the contents of geniposide, baicalin, berberine hydrochloride, and gentian violet hydrochloride was used as the evaluation index. The experimental results were analyzed, and the orthogonal experimental design and results are shown in Table 1.

[0018] Table 1. Overall Scoring Results of Orthogonal Experiments Note: Y1: alcohol precipitation concentration (%), Y2: alcohol precipitation time (h) Combine and mix the three batches of concentrate that have been validated by the water extraction process. Accurately measure nine portions of the above mixed concentrate, each 100 mL, and place them in large beakers for later use.

[0019] According to the experimental design in Table 1, 95% ethanol was diluted to the required alcohol precipitation concentrations (60%, 70%, and 80%). Following the experimental numbers 1-9 in Table 1, the corresponding concentrations of ethanol were added sequentially to the corresponding large beakers containing the concentrated solution. After mixing, the beakers were refrigerated at 4-8°C for alcohol precipitation.

[0020] After the specified alcohol precipitation time in Table 1 has been reached, remove the beaker and filter the contents using double-layered filter paper and a funnel. Collect each portion of the filtered ethanol solution, measure its volume, and refrigerate it at 4–8°C for later use.

[0021] Accurately measure 2 mL of each of the alcohol precipitation purified solutions obtained from the orthogonal experiments 1-9 above, place them in separate 25 mL volumetric flasks, add methanol to the mark, and shake well. Filter through a 0.45 μm microporous membrane, and use the filtrate as the test solution for the alcohol precipitation orthogonal experiment. Accurately pipette 5 μL of each of the above test solutions, inject and determine the concentrations under the optimized chromatographic conditions, record the peak areas of geniposide, baicalin, berberine hydrochloride, and berberine hydrochloride in the chromatogram, and calculate the content of each component.

[0022] The results of the orthogonal experimental analysis of the alcohol precipitation process are shown in Table 2.

[0023] Table 2. Intuitive Analysis Table Note: K: refers to the sum of the experimental scores at the same level for the two factors examined in the alcohol precipitation process (Y1: alcohol precipitation concentration; Y2: alcohol precipitation time). R: mean of the range of a single factor examined. C (blank): blank control factor.

[0024] As can be seen from the intuitive analysis in Table 2, the influence of each factor on the alcohol precipitation effect is ethanol concentration (A) > alcohol precipitation time (B). Based on the comprehensive scoring results in Table 1, the optimal alcohol precipitation process parameters are: add 95% ethanol until the alcohol content reaches 80%, and refrigerate for 36 hours.

[0025] Table 3. Analysis of Variance Table As shown by the analysis of variance in Table 3, the ethanol concentration (A) has a significant effect on the ethanol precipitation effect, while the ethanol precipitation time (B) has no significant effect on the ethanol precipitation effect. Therefore, the ethanol precipitation time can be adjusted to 12~24h.

[0026] Finally, combining the results of orthogonal experiments, intuitive analysis, and analysis of variance, while also considering factors such as manpower, material resources, and time consumption in actual situations, the optimal parameters for the alcohol precipitation process were determined to be: adding 95% ethanol until the alcohol content reaches 80%, and then refrigerating and allowing it to stand for 12 hours.

[0027] Example 2: Preparation of Coptis chinensis detoxifying rinse S1. Raw material preparation and pretreatment Prescription ingredients: Coptis chinensis, Phellodendron chinense, Scutellaria baicalensis, and Gardenia jasminoides are weighed according to the following proportions. The basic prescription is 7.5g of Coptis chinensis, 5.0g of Phellodendron chinense, 5.0g of Scutellaria baicalensis, and 7.5g of Gardenia jasminoides. All ingredients are qualified Chinese medicinal herbs and have been identified as meeting the pharmacopoeia standards.

[0028] Reference standards and reagents: Berberine hydrochloride, berberine hydrochloride, baicalin, geniposide reference standards (purity ≥98%); acetonitrile (chromatographic grade), phosphoric acid (analytical grade), sodium dodecyl sulfate (SDS, analytical grade), 95% ethanol (analytical grade), 20% sodium hydroxide solution, dilute hydrochloric acid solution, sorbic acid (food grade), purified water (meets pharmaceutical requirements).

[0029] Instrument calibration: The high-performance liquid chromatograph (Agilent 1260) underwent a system suitability test in advance to ensure the stability of the chromatographic column and detector; the temperature accuracy of the 98-1-C type digital temperature-controlled electric heating mantle and the DK-98-Ⅱ electric thermostatic water bath was calibrated; the vacuum degree and rotation speed of the R210 type rotary evaporator were checked; the electronic balance (accuracy 0.01g) was calibrated and ready for use; the accuracy of ultrasonic power and time control was confirmed by the KH-250B ultrasonic instrument.

[0030] S2. Preparation process steps Water extraction process: Weigh the medicinal materials according to the prescription, add 14 times the amount of purified water (based on the weight of the medicinal materials), and soak for 2 hours. After soaking, decoct 3 times, 1 hour each time. After decoction, filter the liquid through a double-layer filter cloth, combine the filtrates, cool to room temperature, measure the volume, and store at 4~8℃ for later use.

[0031] Concentration and test sample preparation: The refrigerated water extract filtrate was placed in a rotary evaporator and concentrated under reduced pressure at 80℃ and 0.08~0.09 MPa to 400 mL (concentration of 0.5 g / mL). 2 mL of the concentrated solution was accurately pipetted into a 50 mL volumetric flask, methanol was added to approximately 45 mL, and the mixture was sonicated for 5 min. The volume was then adjusted to the mark with methanol, and the solution was filtered through a 0.45 μm microporous membrane. The filtrate was used as the test sample solution, and the contents of geniposide, baicalin, berberine hydrochloride, and berberine hydrochloride were determined using high-performance liquid chromatography (HPLC).

[0032] Alcohol precipitation process: Mix three batches of validated concentrate, accurately pipette nine portions (100 mL / portion) into a large beaker, dilute 95% ethanol to the preset concentration, add it to the concentrate according to the experimental design ratio, stir well, and refrigerate at 4-8℃ for alcohol precipitation. After alcohol precipitation, filter using a funnel and double-layer filter paper, collect the ethanol liquid, measure the volume, and refrigerate for later use; for scale-up production, directly add 95% ethanol to the alcohol content to 80%, refrigerate and let stand for 12-24 hours, then collect the supernatant.

[0033] Additives: Take the supernatant after alcohol precipitation, adjust the pH to 5.5~6.0 with 20% sodium hydroxide solution and dilute hydrochloric acid solution, and then add 36g of 0.2% sorbic acid.

[0034] Sterilization and dispensing: After sterilizing the prepared liquid, dispense it while hot into sterile containers, seal, and cool to obtain the finished Huanglian Jiedu Rinse Solution. Store the finished product in a cool, dry place. High-density polyethylene bottles are used as the packaging material, with a packaging specification of 100mL / bottle.

[0035] Example 3: Therapeutic effect on a rat model of pressure injury infection (1) Model building and grouping Sixty healthy male SD rats were randomly divided into 5 groups (n=12). The specific groups and treatment methods are as follows: Pressure injury model: A three-stage pressure injury model was established by clamping the skin on the back of rats with magnetic discs (6 hours / day for 3 days).

[0036] Infection model: 0.1 mL of Staphylococcus aureus suspension (10) was inoculated into the pressure wound. 8 (CFU / mL), sealed with dressing for 48 hours to replicate the infected wound model.

[0037] (2) Intervention methods After successful model establishment, each group will undergo intervention according to the following plan, twice daily (9:00 AM and 3:00 PM) for 14 consecutive days: The saline group, iodine solution group, and Coptis chinensis detoxification rinsing group were each given their respective solutions. The solutions were rinsed with a 5mL syringe at a distance of 5-8cm from the wound until no obvious contaminants were visible to the naked eye.

[0038] Model control group and blank control group: No treatment was performed.

[0039] (3) Observation indicators and results Comparison of wound healing rates among different groups of rats at different time points after intervention Table 4. Analysis results of wound healing rate of rats in different time points after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0040] The results showed that on days 3 and 7 of the intervention, the wound healing rate of rats in different intervention groups differed significantly. P <0.05), and there was no statistically significant difference in wound healing rate among different intervention groups on day 14 of intervention ( P>0.05). Pairwise comparisons between groups revealed that on day 3 of intervention, the wound healing rate in the model group was significantly lower than that in the saline group, iodine group, and Coptis chinensis detoxification rinsing group, with statistically significant differences (P<0.05). This indicates that the average wound healing rate in the saline group, iodine group, and Coptis chinensis detoxification rinsing group was comparable, and significantly higher than that in the model group.

[0041] Repeated measures ANOVA and trend changes of wound area and healing rate in rats at different time points after intervention in each group Repeated measures ANOVA was used to analyze the wound healing rate of rats at three time points: day 3, day 7, and day 14 after intervention, and trend graphs were plotted.

[0042] Table 5. Results of repeated measures ANOVA on wound healing rate in rats after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0043] Table 5 shows that the wound healing rate in rats differed statistically significantly across different groups and time periods. P <0.05), and there is an interaction between the between-group effect and the time effect ( P <0.01).

[0044] Figure 1 The results showed that the wound healing rate of the Coptis chinensis detoxification rinsing group increased steadily at all time points, significantly higher than that of the model group, indicating that it has a good effect on promoting wound healing.

[0045] Comparison of bacterial concentrations in rat wounds at different time points after intervention between groups The Kruskal-Wallis H test was used to compare the bacterial concentrations in the wounds of rats in each group on days 1, 3, 7, and 14 of the intervention. Dunnett's T3 test was also used for pairwise comparisons between groups.

[0046] Table 6. Analysis results of bacterial concentration in rat wounds at different time points after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0047] The results showed that the bacterial concentrations in the wounds of rats in different intervention groups differed significantly on days 1, 3, 7, and 14 of the intervention. P <0.01). It can be seen that Huanglian Jiedu irrigation solution can effectively reduce the bacterial concentration in PI-infected wounds and control wound infection by rinsing them.

[0048] Pairwise comparisons between groups showed that on day 1 of intervention, the bacterial concentration in rat wounds was significantly different between the model group and the other three intervention groups. P <0.05), and there were no statistically significant differences among the other three intervention groups. P >0.05); before day 3 of intervention, the model group showed statistically significant differences compared to the iodine group and the Coptis chinensis detoxification rinse group. P <0.05), and there were no statistically significant differences among the other groups. P >0.05); before day 7 of intervention, the model group showed statistically significant differences compared to the iodine group and the Coptis chinensis detoxification rinse group. P <0.05), the difference between the saline group and the Coptis chinensis detoxification rinse group was statistically significant. P <0.05); there were no statistically significant differences among the other groups ( P >0.05); before day 14 of intervention, there was no statistically significant difference between the model group and the saline group, and between the iodine group and the Coptis chinensis detoxification rinsing group. P >0.05), and the differences among the remaining groups were statistically significant. P <0.05).

[0049] Repeated measures ANOVA and trend changes of bacterial concentration in rat wounds at different time points after intervention in each group Repeated measures ANOVA was used to analyze the bacterial concentrations in rat wounds at four time points (days 1, 3, 7, and 14) during the intervention, and trend graphs were plotted.

[0050] Table 7. Results of repeated measures ANOVA on bacterial concentration in rat wounds after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0051] Table 7 shows that the changes in bacterial concentration in rat wounds were statistically significant across different groups and at different time points. P <0.01), and there is an interaction between the between-group effect and the time effect ( P <0.01).

[0052] Figure 2 The results showed that the bacterial concentration in the Coptis chinensis detoxification rinse group decreased rapidly after intervention, comparable to that in the iodine tincture group, and superior to that in the saline group, demonstrating its rapid and long-lasting antibacterial effect.

[0053] Comparison of inflammatory factor expression in wound tissues of rats in different groups after intervention One-way ANOVA was used to analyze the skin tissue of rats in the five groups after day 14 of intervention. IL-1β , IL-6 and TNF-α Concentrations were compared between groups, and pairwise comparisons between groups were performed using the LSD method.

[0054] Table 8. Rats in each group after intervention IL-1β , IL-6 , TNF-α Intergroup analysis results of concentration Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0055] Table 9. Rats in each group after intervention IL-1β , IL-6 , TNF-α Pairwise analysis results between concentration groups Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0056] The results showed that after the intervention, the skin tissue of rats in the blank group, model group, saline group, iodine group, and Huanglian Jiedu rinsing group were significantly different. IL-1β , IL-6 , TNF-α The concentration difference was statistically significant. P <0.01). This indicates that the Coptis chinensis detoxifying rinse prepared in this study has the effect of inhibiting... IL-1β , IL-6 , TNF-α The expression has an anti-inflammatory effect.

[0057] Pairwise comparisons between groups showed that the skin tissue of rats in the blank group, model group, saline group, iodine group, and Huanglian Jiedu rinse group... IL-1β The concentration differences were all statistically significant. P <0.01); Skin tissue of rats in the blank group, model group, saline group, iodine group, and Huanglian Jiedu rinse group IL-6 Except for the iodine solution group, which showed no statistically significant difference in concentration compared to the Coptis chinensis detoxification rinse, (… P >0.05), and the differences among the remaining groups were all statistically significant ( P <0.01); Skin tissue of rats in the blank group, model group, saline group, iodine group, and Huanglian Jiedu rinse group TNF-α There were no statistically significant differences between the concentration model group and the saline group, the iodine group and the Coptis chinensis detoxification rinse group. P >0.05), and the differences among the remaining groups were all statistically significant ( P <0.01).

[0058] Comparison of TLR2 / MyD88 / NF-κB p65 signaling pathway protein expression in wound tissues of rats in different groups after intervention One-way ANOVA was used to compare the expression levels of TLR2, MyD88 and NF-KBp65 proteins in the skin tissue of rats in the five groups after day 14 of intervention. LSD method was also used for pairwise comparisons between groups.

[0059] Table 10. Intergroup analysis of TLR2, MyD88, and NF-κBp65 protein expression levels in rats after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0060] Table 11. Results of pairwise analysis of TLR2, MyD88, and NF-κBp65 protein expression levels in rats after intervention. Note: Intergroup comparison: * P <0.05, ** P <0.01, *** P <0.001.

[0061] The results showed that after the intervention, the expression levels of TLR2, MyD88, and NF-KBp65 proteins in the skin tissue of rats in each group were statistically significant. P <0.01) (e.g. Figure 3As shown). Meanwhile, pairwise comparisons between LSD groups showed that the expression levels of TLR2, MyD88, and NF-KBp65 proteins were not statistically significantly different between the blank group and the Coptis chinensis detoxification rinsing group, and between the model group and the saline group. P >0.05), and the differences among the remaining groups were statistically significant ( P <0.01). Therefore, it can be inferred that Huanglian Jiedu irrigation solution exerts its anti-inflammatory effect on PI-infected wounds by regulating the TLR2 / MyD88 / NF-κB signaling pathway.

[0062] Example 4: Safety Experiment of Coptis chinensis Detoxification Rinse (1) Skin irritation test Four New Zealand rabbits were used, and a damaged skin model was created on their backs. The cleaning agent of this invention and physiological saline were applied to each rabbit's back once daily for seven consecutive days. The skin on the rabbits' backs was scored according to a skin irritation response scoring standard, including erythema and edema. ① Erythema: No erythema = 0 points, barely visible = 1 point, clearly visible = 2 points, severe erythema = 3 points, purplish-red erythema = 4 points; ② Edema: No edema = 0 points, barely visible = 1 point, visible (edge ​​raised above surrounding skin) = 2 points, skin elevation more than 1 mm with clear outline = 3 points, edema elevation more than 1 mm and expanding area = 4 points. The average irritation index = (total erythema score + total edema score) / total number of animals. A score < 0.5 indicates no irritation; a score < 2.0 indicates mild irritation; a score < 6.0 indicates moderate irritation; and a score > 6.0 indicates strong irritation.

[0063] Table 12 Results of skin irritation response scores in experimental rabbits like Figure 4 As shown, no irritation reactions such as erythema or edema were observed on either side of the skin, and the scores were both 0 points, indicating that the product of this application is non-irritating to damaged skin.

[0064] (2) Acute eye irritation test Four rabbits were used. 0.1 mL of the rinsing solution of this invention was instilled into the left eye, and physiological saline was instilled into the right eye as a control. The rabbit eyes were scored according to the rabbit eye irritation scoring system in the Draize test. The final eye stimulation score of each rabbit was obtained by dividing the total score of all observation points by the number of observation points (4 in this experiment). The stimulation level was then graded according to the eye stimulation intensity scoring standard.

[0065] Table 13 Results of Acute Eye Irritation Response Scores in Experimental Rabbits like Figure 5As shown, after 72 hours of observation, except for slight irritation 1 hour after instillation (score 2), there was no irritation reaction at other times (score 0), indicating that this product has extremely low eye irritation and meets the safety requirements for eye rinses.

[0066] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. The use of a Coptis chinensis detoxifying rinse in the preparation of a medicament for the treatment or prevention of infected wounds of the skin and / or mucous membranes.

2. The application according to claim 1, characterized in that, The infected skin wounds include pressure injuries, diabetic foot ulcers, infected burn wounds, and / or infected surgical wounds.

3. A method for preparing a Coptis chinensis detoxifying rinse for use in any one of claims 1 or 2, characterized in that, Includes the following steps: S1: Weigh out the medicinal slices according to the mass ratio of Coptis chinensis: Phellodendron chinense: Scutellaria baicalensis: Gardenia jasminoides in 3:2:2:3 to obtain mixed medicinal materials; S2: Add water to the mixed medicinal materials and decoct to extract, filter for the first time, collect the water extract, and obtain the filtrate; S3: Concentrate the filtrate under reduced pressure to obtain a concentrated solution; S4: The concentrate is subjected to alcohol precipitation, filtered a second time, and the supernatant is collected; S5: Recover the ethanol from the supernatant, adjust the pH, add sorbic acid, and add water to make up to 100 ml; S6: After the liquid solution has been brought to a certain volume, it is sterilized at high temperature and then dispensed to obtain the Coptis chinensis detoxification rinse solution.

4. The preparation method according to claim 3, characterized in that, In step S2, the herbs are soaked in water for 2 hours before decoction, and the decoction is performed 3 times, 1 hour each time, with the amount of water added being 14 times the total weight of the mixed herbs. In step S3, the concentration is carried out under reduced pressure and the temperature is controlled at 80°C. In step S4, 95% ethanol is added to the concentrate to make the final alcohol content of the system reach 80%, and alcohol precipitation is carried out under refrigeration for 12-24 hours. In step S5, the amount of sorbic acid added is 0.1%-0.3% of the final drug solution mass.

5. The Coptis chinensis detoxification rinse prepared by the preparation method according to any one of claims 3 or 4.

6. The Coptis chinensis detoxifying rinse according to claim 5, characterized in that, The Coptis chinensis detoxification rinse contains geniposide, baicalin, berberine hydrochloride, and berberine hydrochloride as active ingredients.

7. The Coptis chinensis detoxifying rinse according to claim 5, characterized in that, The Coptis chinensis detoxifying rinse is used to inhibit pro-inflammatory factors. IL-1β , IL-6 and TNF-α The expression of key proteins in the TLR2 / MyD88 / NF-κB inflammatory pathway was downregulated to reduce bacterial load in the wound and promote wound healing.

Images