A traditional Chinese medicine composition for treating diabetic foot ulcers, a preparation method thereof and application thereof

Abstract

The application discloses a traditional Chinese medicine composition for treating diabetic foot ulcers and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine, wherein the traditional Chinese medicine composition comprises the following components in parts by weight: 2-10 parts of wind-preventing plant, 2-10 parts of Qianghuang, 20-40 parts of mulberry branch, 10-20 parts of Chinese gall, 2-10 parts of golden yellow, 2-10 parts of white peony root, 10-30 parts of rhubarb, 2-10 parts of forsythia, and 5-15 parts of honeysuckle. The traditional Chinese medicine composition has a good treatment effect on diabetic foot ulcers, and effectively solves the problem of poor curative effect in the prior art.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating diabetic foot ulcers and a preparation method and application thereof. BACKGROUND

[0002] Diabetic foot ulcer is a chronic complication of diabetic patients caused by long-term high blood sugar, mainly manifested as full-thickness skin damage of the foot. This disease is particularly common in the elderly, and its incidence significantly increases with age. Diabetes and its complications not only seriously affect the quality of life of patients, but also bring a heavy economic burden to families and society.

[0003] Diabetic foot is mainly caused by diabetic microangiopathy, neuropathy and bacterial infection. Multiple factors jointly cause the complex pathology of diabetic foot, and the complex pathology is the reason why diabetic foot ulcer is difficult to heal.

[0004] At present, the treatment strategy for diabetic foot ulcer (DFU) is blood glucose control, which can be achieved through insulin programs (such as insulin glargine, insulin aspart) or oral hypoglycemic drugs, including metformin, sodium-glucose cotransporter 2 (SGLT2) inhibitors (such as empagliflozin) and glucagon-like peptide-1 (GLP-1) receptor agonists (such as liraglutide). Wound debridement needs to follow standardized assessment methods, such as Wagner classification, ankle-brachial index (ABI) and transcutaneous oxygen partial pressure (TcPO2), to determine the depth of ulcer, the range of infection and the burden of ischemia. According to the characteristics of the wound, different debridement methods are selected, including surgical sharp debridement, enzymatic debridement, autolytic debridement, mechanical debridement and biological debridement. Infection management includes clinical manifestations, microbiology and imaging evaluation, and then according to the severity of infection and pathogenic bacteria, a systemic antibiotic treatment plan (such as amoxicillin, vancomycin hydrochloride and daptomycin) is developed, supplemented by antibacterial dressings or local antibiotic treatment. Finally, reduced pressure treatment is a key intervention measure, aiming to redistribute the pressure on the foot and optimize local perfusion, thereby accelerating wound healing and preventing recurrence.

[0005] In recent years, traditional Chinese medicine has gradually been valued in the treatment of diabetic foot ulcers. Traditional Chinese medicine can regulate the balance of yin and yang of the whole body, and promote the metabolism of sugar, fat and protein. A large number of clinical studies have shown that traditional Chinese medicine has unique advantages in the treatment of diabetic foot ulcers, such as multi-component, multi-target and multi-pathway comprehensive action, and fewer side effects.

[0006] "Xiaoke disease", i.e. diabetes, diabetic patients belong to heat asphyxia, especially biliary stasis, manifested as restlessness, weakness, less food, irregular bowel movements, abdominal pain, red blisters and foam on the tongue, and further development of diabetic retinopathy, neuropathy, diabetic foot ulcer and other complications.

[0007] Currently, there are numerous traditional Chinese medicine products on the market for diabetic foot ulcers, but the efficacy and safety of most of these products still require further verification. Therefore, developing a traditional Chinese medicine product that is effective, safe, and easy to use is of great significance for improving the quality of life for patients with diabetic foot ulcers. Summary of the Invention

[0008] In view of the above-mentioned shortcomings in the prior art, the present invention provides a traditional Chinese medicine composition for treating diabetic foot ulcers, its preparation method and application. The traditional Chinese medicine composition has a good therapeutic effect on diabetic foot ulcers and effectively solves the problem of poor efficacy in the prior art.

[0009] To achieve the above objectives, the technical solution adopted by the present invention to solve its technical problem is as follows:

[0010] A traditional Chinese medicine composition for treating diabetic foot ulcers comprises the following components in parts by weight: 2-10 parts of Saposhnikovia divaricata, 2-10 parts of Notopterygium incisum, 20-40 parts of Morus alba twigs, 10-20 parts of Gallnut, 2-10 parts of Phellodendron chinense, 2-10 parts of Angelica dahurica, 10-30 parts of Rheum palmatum, 2-10 parts of Forsythia suspensa, and 5-15 parts of Lonicera japonica.

[0011] Further, it includes the following components in parts by weight: Saposhnikovia divaricata 4-7 parts, Notopterygium incisum 4-7 parts, Morus alba twigs 25-35 parts, Gallnut 12-17 parts, Phellodendron chinense 4-7 parts, Angelica dahurica 4-7 parts, Rheum palmatum 17-24 parts, Forsythia suspensa 4-7 parts, and Lonicera japonica 8-12 parts.

[0012] Furthermore, it includes the following components in parts by weight: 4 parts of Saposhnikovia divaricata, 7 parts of Notopterygium incisum, 35 parts of Morus alba twigs, 17 parts of Galla chinensis, 7 parts of Phellodendron chinense, 7 parts of Angelica dahurica, 17 parts of Rheum palmatum, 4 parts of Forsythia suspensa, and 8 parts of Lonicera japonica.

[0013] Furthermore, it includes the following components in parts by weight: 7 parts of Saposhnikovia divaricata, 4 parts of Notopterygium incisum, 25 parts of Morus alba twigs, 12 parts of Galla chinensis, 4 parts of Phellodendron chinense, 4 parts of Angelica dahurica, 24 parts of Rheum palmatum, 7 parts of Forsythia suspensa, and 12 parts of Lonicera japonica.

[0014] Furthermore, it includes the following components in parts by weight: 5 parts of Saposhnikovia divaricata, 5 parts of Notopterygium incisum, 30 parts of Morus alba twigs, 15 parts of Galla chinensis, 5 parts of Phellodendron chinense, 5 parts of Angelica dahurica, 20 parts of Rheum palmatum, 5 parts of Forsythia suspensa, and 10 parts of Lonicera japonica.

[0015] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: preparing the effective components obtained by decocting and extracting each raw material.

[0016] Furthermore, the prepared dosage form is a topical dressing.

[0017] Furthermore, the specific preparation method is as follows: the above drugs are soaked in water, then decocted, filtered, the filtrate is collected and concentrated, and pharmaceutically acceptable excipients are added to obtain the final product.

[0018] Furthermore, the amount of water added should be 9-11 times the weight of the raw material, the soaking time should be 20-50 minutes, and the decoction should be performed 2-3 times, with each decoction lasting 1-1.5 hours.

[0019] The above-mentioned traditional Chinese medicine composition is used in the preparation of topical dressings for treating diabetic foot ulcers.

[0020] The beneficial effects of this invention are as follows:

[0021] In this invention, mulberry twigs have the effects of dispelling wind and dampness, clearing the meridians, benefiting the joints, and promoting the flow of water and qi. They can guide the medicines to the limbs, especially the lower limbs. In this application, mulberry twigs enhance the body's blood circulation, relieve the core pathogenesis of poor blood circulation in the limbs, and thus promote ulcer repair. In this application, mulberry twigs are used as the principal medicine to play the main therapeutic role.

[0022] Phellodendron bark has the effects of clearing heat and drying dampness, purging fire and detoxifying, and reducing deficiency heat. It can promote the excretion of dampness in the lower limbs of patients and accelerate ulcer repair. Angelica dahurica has the effects of drying dampness and stopping leukorrhea, and reducing swelling and draining pus. It can promote the discharge of ulcer abscesses. Its aromatic and penetrating properties can guide the medicine to penetrate the skin surface and increase the efficacy of local medication. Phellodendron bark and Angelica dahurica work together to enhance the ability to dry dampness. Forsythia and honeysuckle both have the effects of clearing heat and detoxifying, and dispersing wind-heat. Forsythia focuses on detoxifying and dispersing nodules, while honeysuckle focuses on cooling blood and detoxifying. The two work together with Phellodendron bark to enhance the power of clearing heat and detoxifying. They play a therapeutic role in the local redness, swelling, burning, and thick pus of ulcers. All of the above are assistant medicines.

[0023] Both Saposhnikovia divaricata and Notopterygium incisum have the effects of dispelling wind and relieving exterior symptoms, eliminating dampness and relieving pain. They can assist the principal drug in unblocking the meridians and enhancing the analgesic effect. Gallnut has the effects of astringing sweat and stopping bleeding, and astringing dampness and healing sores. Its sour and astringent properties can reduce the exudation of fluid from the wound and promote wound healing. All of the above are adjuvant drugs.

[0024] Rhubarb has the effects of clearing heat and purging fire, cooling blood and detoxifying, and removing blood stasis and promoting menstruation. It can cleanse the body of blood stasis and dampness, promote the excretion of necrotic tissue, and when combined with mulberry branches, it can exert the effects of clearing the meridians and removing blood stasis and heat, and play a therapeutic role in the pathogenesis of blood stasis and internal accumulation of heat and toxins.

[0025] The above composition treats diabetic foot ulcers by clearing heat and detoxifying, stopping bleeding and promoting tissue regeneration, reducing swelling and relieving pain, and cooling blood and dispersing blood stasis. The drugs in the composition work synergistically and complement each other, which can better promote wound healing, angiogenesis, and antibacterial and bacteriostatic effects, significantly improving the treatment effect of diabetic foot ulcers. It has a good therapeutic effect on diabetic foot ulcers throughout the entire cycle. Attached Figure Description

[0026] Figure 1 This is a diagram showing the results of wound repair. Figure 1 A shows photos of the wound repair process in each group; Figure 1 B is a statistical chart of wound healing rate;

[0027] Figure 2 HE staining and statistical analysis of scar length and reepithelialization rate;

[0028] Figure 3 Statistics on Masson staining and its collagen deposition;

[0029] Figure 4 Statistical analysis of immunofluorescence and fluorescence intensity of Staphylococcus aureus;

[0030] Figure 5 This is a target intersection diagram of a novel traditional Chinese medicine compound and diabetic foot ulcers.

[0031] Figure 6 This is a diagram showing the results of protein-protein interactions, where... Figure 6 A is the PPI chart; Figure 6 B represents the Network diagram;

[0032] Figure 7 Upset image;

[0033] Figure 8 The image shows the results of pathway enrichment analysis for GO Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Figure 8 A is the GO enrichment analysis plot. Figure 8 B is a KEGG pathway enrichment analysis diagram;

[0034] Figure 9 This is a drug-component-target-disease network diagram.

[0035] Figure 10 A thermal image of molecular docking;

[0036] Figure 11 This is a schematic diagram simulating the docking of different molecules, where, Figure 11 A is a schematic diagram of the docking simulation between TNF and quercetin molecules. Figure 11 B is a schematic diagram of the molecular docking simulation between AKT1 and kaempferol. Figure 11 C is a schematic diagram of the molecular docking simulation of EGFR and luteolin;

[0037] Figure 12 Images of the foot wounds of patient 1 before and after treatment; among them, Figure 12 A shows the foot wound of patient 1 before admission. Figure 12 Image B shows the patient's foot wound on the 7th day after admission. Figure 12 C shows the patient's foot wound on the 11th day after admission.Figure 12 D shows the foot wound of the patient 20 days after admission;

[0038] Figure 13 Images of the foot wounds of patient 2 before and after treatment are shown. Figure 13 A shows the foot wound of patient 2 before admission. Figure 13 Image B shows the patient's foot wound on the 7th day after admission. Figure 13 C shows the patient's foot wound on the 11th day after admission. Figure 13 D shows the foot wound of the patient 20 days after admission;

[0039] Figure 14 Line graph showing the changes in healing rates for Patient 1 and Patient 2. Detailed Implementation

[0040] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention; that is, the described embodiments are merely some embodiments of the invention, and not all embodiments.

[0041] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0042] It should be noted that relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes the element.

[0043] The features and performance of the present invention will be further described in detail below with reference to the embodiments and accompanying drawings.

[0044] Example 1

[0045] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 5 parts of Saposhnikovia divaricata, 5 parts of Notopterygium incisum, 30 parts of Morus alba twigs, 15 parts of Galla chinensis, 5 parts of Phellodendron chinense, 5 parts of Angelica dahurica, 20 parts of Rheum palmatum, 5 parts of Forsythia suspensa, and 10 parts of Lonicera japonica.

[0046] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution, mix well, and obtain the external dressing.

[0047] Example 2

[0048] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 7 parts of Saposhnikovia divaricata, 4 parts of Notopterygium incisum, 25 parts of Morus alba twigs, 12 parts of Galla chinensis, 4 parts of Phellodendron chinense, 4 parts of Angelica dahurica, 24 parts of Rheum palmatum, 7 parts of Forsythia suspensa, and 12 parts of Lonicera japonica.

[0049] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution, mix well, and obtain the external dressing.

[0050] Example 3

[0051] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 2 parts of Saposhnikovia divaricata, 10 parts of Notopterygium incisum, 40 parts of Morus alba twigs, 20 parts of Galla chinensis, 10 parts of Phellodendron chinense, 10 parts of Angelica dahurica, 10 parts of Rheum palmatum, 2 parts of Forsythia suspensa, and 5 parts of Lonicera japonica.

[0052] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution, mix well, and obtain the external dressing.

[0053] Example 4

[0054] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 10 parts of Saposhnikovia divaricata, 2 parts of Notopterygium incisum, 20 parts of Morus alba twigs, 10 parts of Galla chinensis, 2 parts of Phellodendron chinense, 2 parts of Angelica dahurica, 30 parts of Rheum palmatum, 10 parts of Forsythia suspensa, and 15 parts of Lonicera japonica.

[0055] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution to obtain an external dressing.

[0056] Example 5

[0057] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 4 parts of Saposhnikovia divaricata, 7 parts of Notopterygium incisum, 35 parts of Morus alba twigs, 17 parts of Galla chinensis, 7 parts of Phellodendron chinense, 7 parts of Angelica dahurica, 17 parts of Rheum palmatum, 4 parts of Forsythia suspensa, and 8 parts of Lonicera japonica.

[0058] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution to obtain an external dressing.

[0059] Comparative Example 1

[0060] A traditional Chinese medicine composition for treating diabetic foot ulcers, comprising the following components in parts by weight: 5 parts cinnamon twig, 5 parts notopterygium root, 30 parts mulberry twig, 15 parts dried plum, 5 parts phellodendron bark, 5 parts chuanxiong rhizome, 20 parts rhubarb, 5 parts forsythia fruit, and 10 parts honeysuckle flower.

[0061] The preparation method of the above-mentioned traditional Chinese medicine composition for treating diabetic foot ulcers includes the following steps: Take each medicinal material, dry it, and finely pulverize it (95% passing through an 80-mesh sieve); Place the above medicines in a container, add water to soak them, and decoct them three times. For the first decoction, add 10 times the weight of the raw materials of water, soak for 30 minutes to allow the medicinal materials to fully absorb water and swell, then bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the second decoction, add 10 times the weight of the raw materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, and collect the filtrate; for the third decoction, add 10 times the weight of the medicinal materials of water, bring to a boil over high heat and simmer over low heat for 1 hour, filter, collect the filtrate, combine the filtrates, and concentrate the filtrate under reduced pressure using a rotary evaporator until the relative density is 1.2. Then add petrolatum to the filtrate, with a volume-to-mass ratio of 1 mL: 0.5 g for the concentrated solution to obtain an external dressing.

[0062] In Examples 1-5 and Comparative Example 1 above, the relative density of the concentrate was measured according to the specific gravity bottle method in the 2020 edition of the Chinese Pharmacopoeia.

[0063] Test case

[0064] I. Animal Experiments

[0065] 1. Selection of experimental mice: Male C57BL / 6 mice (from Sichuan Vital River Laboratory Animal Technology Co., Ltd.) aged 5-6 weeks and weighing 18g-22g were selected and conditioned for 7 days. Before modeling, the weight was measured, the tail vein blood was detected by a blood glucose meter, and the blood glucose level of the blank serum for modeling was recorded.

[0066] 2. Construction of a diabetic mouse model: 1% streptozotocin (STZ) was dissolved in 0.1 mol / L citrate-sodium citrate buffer (0.1 mol / L, pH=4.5). The dissolved solution should be used within 30 minutes and stored on ice in the dark. Starting from the second week of feeding, mice were fasted for 16 hours. The control group received intraperitoneal injections of citrate buffer for five consecutive days, while the diabetic group received STZ injections for five consecutive days at a dose of 60 mg / kg / day, on a high-sugar, high-fat diet. In the first week after all injections were completed, tail vein blood samples were measured three times consecutively using a blood glucose meter. A random fasting blood glucose concentration higher than 11.1 mmol / L was considered a successful model.

[0067] 3. Construction of a diabetic mouse injury model: After successfully establishing a diabetic mouse model, wound creation was performed. Mice were anesthetized with 50 mg / kg pentobarbital. Hair was removed from the backs of the mice using a shaving tool and depilatory cream, followed by routine disinfection. Under aseptic conditions, a 1×1 cm² wound was created on the back skin of the mice using a sterile scalpel and punch. The wounds were disinfected with clean gauze to prevent infection. Each wound was inoculated with 200 μL of a 1×10⁻⁶ solution. 8 CFU of Staphylococcus aureus was administered subcutaneously in four doses, 50 μL each time. The wound was observed 24 hours later. If yellow purulent discharge appeared on the wound, the surrounding skin temperature was slightly elevated, the skin was red, the animals were slightly lethargic, and had a poor appetite, then the pus smears of the animals in each group were randomly selected for microscopic examination. If a large number of Staphylococcus aureus were observed or the isolation and culture results were Staphylococcus aureus, then the model was considered to have been successfully established.

[0068] 4. Experimental grouping and drug administration: Fifteen mice were randomly divided into a control group (100 μg vancomycin hydrochloride + 5 mL PBS) and a control group (0.1 g of the drug in control group + 5 mL PBS) and an experimental group (0.1 g of the drug in example 1 + 5 mL PBS) and an experimental group (5 mice per group). The drugs were applied locally to the wound surface twice a day for 9 days.

[0069] 5. Wound Healing Rate Statistics: The experiment lasted for 9 days. To observe the development of chronic diabetic wounds and assess treatment effectiveness, digital images were acquired on the day of surgery, and subsequently every other day after wound formation. Standardized calibration was used to measure wound area, and ImageJ software was used to calculate the wound closure rate. The data were then substituted into the formula: Wound Healing Rate = (Initial Wound Area - Wound Area on Current Day) / Initial Wound Area × 100%. The wound healing rate on day 9 after modeling was calculated, and the time for eschar and pus to fall off, wound color, amount of wound exudate, and granulation tissue growth were observed and recorded.

[0070] 6. Record detailed measurement data for each mouse, enter the data into an Excel spreadsheet, and then perform statistical analysis using Graphpadprism 10.0 software. Import the measured data into the analysis software for further data processing and analysis to assess the degree of wound healing and treatment effectiveness. See the attached results for details. Figures 1-4 .

[0071] Figure 1 A shows photos of the wound repair process in each group. Figure 1 B is a statistical chart of wound healing rate. It can be seen that compared with the vancomycin hydrochloride group, the drugs in the comparative group and the example group have a better repair effect on the wound. After 9 days, the wound is basically healed. Although the vancomycin hydrochloride group has a certain repair effect, it is far less than that of the comparative group and the example group. This is mainly because vancomycin hydrochloride only has a bactericidal effect and can only inhibit wound infection, but it does not promote wound healing, epithelial cell regeneration, or collagen deposition, resulting in a slower wound healing speed. The repair effect of the example group is slightly higher than that of the comparative group. It is speculated that this is because the adjustment of the medicinal materials in the formula has strengthened the interaction between the components and improved the effect. Therefore, the efficacy of the drugs in the comparative group is not as good as that of the drugs in the example group. In summary, the medicinal materials in the composition of this application have a certain interaction, which makes it have a better promoting effect on the repair of diabetic foot ulcer wounds.

[0072] Figure 2 HE staining of wound skin and statistical analysis of scar length and reepithelialization rate; Figure 3 Masson staining of wound skin and statistical analysis of collagen deposition. Figure 4 The immunofluorescence and fluorescence intensity statistics of Staphylococcus aureus showed that the drug in Example 1 was significantly better than other groups in terms of wound regeneration, epithelialization, and promotion of collagen regeneration, further proving that the drug in this application has the effect of promoting the accelerated healing of chronic diabetic wounds.

[0073] II. Network Pharmacological Analysis for the Treatment of Diabetic Foot Ulcers

[0074] 1. Research Methods

[0075] 1.1 Screening of active ingredients and related targets.

[0076] Taking the drug in Example 1 as an example, the compound components and targets of Angelica dahurica, rhubarb, Saposhnikovia divaricata, Phellodendron chinense, Lonicera japonica, Forsythia suspensa, Notopterygium incisum, Morus alba twigs, and Galla chinensis were retrieved from the TCMSP database (www.tcmsp-e.com). The component screening condition was DL≥0.18. All compounds were entered into the SwissTargetPrediction website (www.swisstargetprediction.ch) to predict their targets. Then, all targets were converted into the gene names corresponding to the target proteins using the UniProt database (www.uniprot.org / uniprotkb), and the biological species selected was homosapiens.

[0077] 1.2 Target screening for diabetic foot ulcers

[0078] Using "Diabetic foot ulcer" as the keyword, a search for disease targets was conducted in the GeneCards database (www.genecards.org). All targets were converted to their corresponding gene names using the uniprot database, and the selected biological species was *Homo sapiens*. The disease-related targets obtained from this database were then used as targets for treating diabetic foot ulcers.

[0079] 1.3 Acquisition of relevant target points for drug treatment of diabetic foot ulcers in Example 1

[0080] The obtained drug targets (score ≥ 0.3) and targets for diabetic foot ulcers (score ≥ 10) are entered into the Venn diagram creation website Venny 2.1 (https: / / bioinfogp.cnb.csic.es / tools / venny / ). The intersection of the two is taken to obtain the intersection targets, which can be considered as drug targets for the treatment of diabetic foot ulcers.

[0081] 1.4 PPI Network Construction

[0082] Protein-protein interactions are crucial for cells to perform their physiological functions. The String database (https: / / string-db.org / cgi / input.pl) is an online analysis platform for protein-protein interactions. The relevant targets for treating diabetic foot ulcers obtained above were input into the String database to construct a protein-protein interaction (PPI) network. The biological species was set to "Homo sapiens." Based on the specific network connections, a confidence level of 0.4 was selected, with other parameters set to default, resulting in a suitable PPI network graph. This network has 45 nodes, 282 edges, and an average degree of 12.5. Figure 6 A). Next, download the relevant CSV file and import it into Cystoscape 3.8.0. Use the Network Analyzer tool to perform topology analysis, taking the intersection of the network topology parameters Degree, EPC, MCC, and MNC respectively, and then use RStudio to draw the Upset diagram. Figure 7 Five targets were selected as core targets (Table 1).

[0083] 1.5 Enrichment analysis of GO and KEGG

[0084] The relevant targets for treating diabetic foot ulcers obtained in section 1.3 were input into the Metascape platform (https: / / metascape.org / gp / index.html# / main / step1), with P < 0.01 set and the species limited to "Homo sapiens". GO function and KEGG pathway enrichment analyses were performed, and the obtained data were finally visualized using the Bioinformatics platform (https: / / www.bioinformatics.com.cn / ).

[0085] 1.6 Drug-Compound-Target-Disease Network Diagram

[0086] To better understand the complex interactions between drugs, ingredients, diseases, and their corresponding targets, a drug-ingredient-target-disease network diagram was constructed using Cytoscape 3.8.0. Then, its built-in tool, NetworkAnalyzer, was used to calculate the network topology parameters of the drug's active ingredients and targets. The top 5 active ingredients in terms of network topology parameter Degree were identified as potentially key components in the drug treatment of diabetic foot ulcers in Example 1.

[0087] 1.7 Molecular docking

[0088] Molecular docking was performed between the core target molecules screened in the 1.4 PPI network (Table 1) and the top 5 active ingredients in terms of network topology parameter Degree screened in the 1.6 network (Table 2). SDF format files of the above active ingredients were downloaded from the PubChem database, then imported into Chem3D software for energy minimization processing. After processing, the files were exported as mol2 format files, and high-resolution 3D structures of the core proteins containing ligand molecules and obtained using X-ray diffraction were obtained from protein databases. Using AutoDockTools 1.5.6 and PyMOL 2.3.4 software, water molecules were removed from key target sites, ligands and acceptors were separated, polar hydrogen was added, Gasteiger charges were calculated, AD4 type was assigned, and flexible bonds of small molecule ligands were set to rotatable. The data was then exported as a pdbqt file. Finally, molecular docking was performed using AutoDock Vina 1.1.2, and the binding energy results were plotted as a heatmap using Prism software. A lower binding energy indicates a darker color, signifying higher binding activity between the target and ligand. PyMOL 2.3.2 software was used to visualize the molecular docking models with the best binding energies for three different proteins.

[0089] 2. Results

[0090] 2.1 Active ingredients and related targets of the drug in Example 1

[0091] After summarizing the database and removing duplicates, the following results were obtained: Angelica dahurica yielded 45 compound components and 1774 targets; Rheum palmatum yielded 63 compound components and 1730 targets; Saposhnikovia divaricata yielded 27 compound components and 1773 targets; Phellodendron chinense yielded 69 compound components and 2896 targets; Lonicera japonica yielded 89 compound components and 3046 targets; Forsythia suspensa yielded 88 compound components and 2244 targets; Notopterygium incisum yielded 45 compound components and 2448 targets; Morus alba twigs yielded 16 compound components and 582 targets; Galla chinensis yielded 1 compound component and 18 targets. Targets with a score ≥0.3 were included, and duplicates were removed, resulting in a total of 291 effective components and 185 targets.

[0092] 2.2 Collection of targets related to diabetic foot ulcers

[0093] Targets for diabetic foot ulcers were obtained from the Genecards database, and targets with a score ≥10 were included, resulting in 591 relevant targets for diabetic foot ulcers.

[0094] 2.3 Targets for the treatment of diabetic foot ulcers

[0095] The 185 drug targets from Example 1 and the 591 drug targets related to diabetic foot ulcers were selected. Taking the intersection of these two groups yielded 47 drug targets from Example 1 for treating diabetic foot ulcers. These 47 targets can be considered potential targets for the drug treatment of diabetic foot ulcers in Example 1. See details below. Figure 5 .

[0096] 2.4 PPI Network Analysis

[0097] The relevant targets and PPI network analysis of the drug treatment for diabetic foot ulcers in Example 1 are shown in [reference needed]. Figure 6 A, Figure 6 B. This network has 45 nodes, 282 edges, and an average degree of 12.5. Next, its built-in tool, NetworkAnalyzer, was used to calculate the network topology parameters of the active pharmaceutical ingredient and targets in Example 1. The top 5 targets with the highest Degree in the network topology parameters were identified as potentially the core targets for the drug treatment of diabetic foot ulcers in Example 1. See Table 1. These targets can be considered the core targets for the drug treatment of diabetic foot ulcers in Example 1. Figure 7 Based on the network topology parameters Degree, EPC, MCC, and MNC, the intersections are taken respectively, and the Upset graph is drawn using RStudio.

[0098] Table 1: Core targets for drug treatment of diabetic foot ulcers in this application

[0099] Core Target TNF AKT1 EGFR ESR1 PTGS2

[0100] 2.5 GO and KEGG enrichment analysis

[0101] Intersection target data were imported into the Metascape data platform for GO and KEGG enrichment analysis. Enrichment yielded 108 KEGG pathways, and GO analysis identified 2262 biological processes (BP), 62 cellular components (CC), and 214 molecular functions (MF). The main GO and KEGG analysis results were visualized using the MicroBioinformatics platform, as shown below. Figure 8 A Figure 8B. As can be seen, the drug targets for treating diabetic foot ulcers in Example 1 mainly regulate biological processes such as catalytic activity, lipid metabolism, immune system processes, insulin secretion, gene expression, cellular responses to oxidative stress, small molecule binding, protein localization, and anion binding. The cellular components affected include cell development, catabolism, regulation of cellular components and tissues, responses to cytokines, signal release, and regulation of biological quality, positive regulation of cell communication, regulation of responses to stimuli, responses to organic matter, regulation of signal transduction, and responses to stress. The related pathways involve signaling pathways such as the HIF-1 signaling pathway, PI3K-Akt signaling pathway, VEGF signaling pathway, AGE-RAGE signaling pathway in diabetic complications, MAPK signaling pathway, TNF signaling pathway, AMPK signaling pathway, and Jak-STAT signaling pathway.

[0102] 2.6 Drug-Compound-Target-Disease Network Diagram

[0103] A drug-compound-target-KEGG pathway network diagram was drawn using Cytoscape 3.8.0. (See attached image.) Figure 9 The network contains 328 nodes and 1220 edges. This graph visually demonstrates that the drug in Example 1 may exert its therapeutic effect on diabetic foot ulcers through multiple components and targets. The network topology analysis of the obtained drug-compound-target-disease network graph was performed using the built-in Network Analyzer of Cytoscape 3.8.0. Then, the network topology parameters of the active ingredients and targets of the drug in Example 1 were calculated using its built-in tool, Network Analyzer. The top 10 active ingredients in terms of network topology parameter Degree were identified as potentially key components in the treatment of diabetic foot ulcers using the drug in Example 1. These components are likely key components that play a role in the treatment of diabetic foot ulcers using the drug in Example 1, as shown in Table 2.

[0104] Note: Figure 7 In the network, green represents the active ingredient, orange represents the drug's target for the disease, and blue represents the KEGG pathway.

[0105] Table 2: Key Ingredients

[0106] Traditional Chinese Medicine Active Ingredient Degree Quercetin 95 Kaempferol 92 Myricetin 78 Apigenin 58 High Glycosides 54 Oleanolic Acid 37 Stigmasterol 25 Beta-sitosterol 23 ZINC04081604 19 Figure 10 16

[0107] 2.7 Molecular docking

[0108] The core drug compounds obtained from Example 1—quercetin, kaempferol, myricetin, luteolin, apigenin, TNF (PDBID: AF-P50592-F1), AKT1 (PDBID: AF-P31749-F1), EGFR (PDBID: AF-P00533-F1, ESR1 (PDBID: 1A52)), and PTGS2 (PDBID: AF-P35354-F1)—were subjected to molecular docking. The binding energies between the core compounds and the core targets were calculated to predict their binding activity. A binding energy below 0 indicates that the two molecules can spontaneously bind, and a higher negative binding energy indicates a more stable conformation. Generally, a binding energy < -5.0 kcal·mol⁻¹ is considered acceptable. -1 This indicates good binding ability; see Figure 11 As can be seen from the figure, most of the docking results are less than 0, and the binding energy of all 25 docking results is < -5.0 kcal·mol⁻¹. -1 It is evident that the core components and core targets can bind well to each other, demonstrating the effective binding of the core components of the drug in Example 1 to the core targets. This reflects the feasibility of the drug's therapeutic effect on diabetic foot ulcers in Example 1. Finally, this model was used to demonstrate the interaction between the two using PyMOL 2.3.2. Here, we present molecular docking simulations showing the best binding energies of three different proteins (TNF and quercetin, AKT1 and kaempferol, EGFR and luteolin). Figure 11 A, Figure 11 B Figure 12 C.

[0109] 3. Summary

[0110] In summary, network pharmacology and molecular docking methods were used to predict the material basis and potential mechanism of the drug in Example 1 for treating diabetic foot ulcers. It was found that the drug in Example 1 may act on potential targets related to TNF, AKT1, EGFR, and ESR1 through its active ingredients such as quercetin, kaempferol, myricetin, luteolin, apigenin, and hyperoside, affecting signaling pathways including HIF-1, PI3K-Akt, VEGF, AGE-RAGE, MAPK, TNF, AMPK, and Jak-STAT. This intervention interferes with biological processes such as catalytic activity, lipid metabolism, immune system processes, insulin secretion, gene expression regulation, cellular responses to oxidative stress, small molecule binding, protein localization, and anion binding, thereby treating diabetic foot ulcers. This demonstrates the multi-component, multi-pathway, and multi-target characteristics of the drug in this application for treating diabetic foot ulcers.

[0111] III. Clinical Trials

[0112] For example, two clinical patients: Patient 1 is a male, aged 69; Patient 2 is a male, aged 68.

[0113] like Figure 12 As shown, a deep ulcer lesion of approximately 10 cm² was visible on the heel of patient 1 at the initial examination. Figure 12 A) The wound is irregular in shape, with red and swollen edges, and the base is covered with a large amount of yellowish-green necrotic tissue and purulent secretions, indicating that the wound is in the infection stage, the local tissue activity is poor, and the wound microenvironment is harsh.

[0114] After 7 days of traditional Chinese medicine intervention treatment in the example group () Figure 12 (B) The wound showed significant improvement locally, with necrotic tissue largely removed, exudate reduced, and inflammatory redness and swelling significantly alleviated. The wound edges gradually became neat, and fresh, moist, red granulation tissue could be seen growing from the bottom of the wound, indicating that the wound had successfully transitioned from the infection phase to the inflammation resolution phase and the initial stage of the proliferative phase.

[0115] Continue intervention until day 11 ( Figure 12 (C) The wound healing process accelerated, the granulation tissue continued to fill and was bright red, with a dense texture, and signs of epithelialization appeared at the wound edges. The exudate almost disappeared, and the wound area shrank by more than 50% compared to the time of admission. No adverse reactions such as local allergies or irritation occurred during the period, demonstrating the good safety and tolerability of this traditional Chinese medicine preparation in clinical use.

[0116] Until the 20th day ( Figure 13 (D) The wound has healed further, with abundant granulation tissue on the surface, and the epithelium at the wound edges gradually covering it. The depth and area of ​​the wound have decreased further compared to before, and the healing rate exceeds 80%. At the same time, the local skin color has returned to normal, with no obvious exudation or infection, indicating that the wound has entered the maturation / remodeling stage.

[0117] like Figure 13 As shown, Patient 2 was admitted to the hospital in the early stages ( Figure 13 A) A deep ulcer with an area of ​​about 10 cm² is visible on the heel. The ulcer is irregular in shape, with a large amount of dark brown and yellowish-green necrotic tissue at the base, accompanied by purulent discharge. The wound edges are obviously red and swollen, and the surrounding skin is hardened, indicating that the wound is in a severe infection stage and the local tissue activity is extremely poor.

[0118] After 7 days of treatment with vancomycin hydrochloride ( Figure 13 (B) Necrotic tissue has partially sloughed off, but a large amount of yellow purulent exudate is still visible, and the wound microenvironment has not been significantly improved. Although the redness and swelling at the wound edges have slightly subsided, the overall condition of the wound remains in the infection stage and early inflammatory response stage, with no obvious granulation tissue formation observed.

[0119] Continue treatment until day 11 ( Figure 13(C) Wound exudate has decreased slightly compared to before, and the wound edges are becoming smoother, but the newly formed granulation tissue is still relatively thin, pale red in color, and soft in texture, indicating insufficient granulation tissue activity. Local wound repair is progressing slowly, with no significant change in area, and inflammation control remains unsatisfactory.

[0120] Until the 20th day ( Figure 14 (D) Although wound exudation further decreased and some areas showed adhesion-like tissue coverage, the overall healing progress still lagged behind that of patients who underwent drug intervention in Example 1. The granulation tissue remained pale in color, with no obvious signs of epithelialization, and no significant contraction was observed at the wound edges, indicating that the wound healing rate in the control group was slower within the same time period, suggesting that conventional dressing changes have limited effectiveness in promoting repair.

[0121] ​ The results show that the wound healing rates of patients treated with the drug intervention in Example 1 were significantly higher on days 7, 11, and 20 than those treated with vancomycin hydrochloride on the same days. This trend indicates that the drug intervention in Example 1 can effectively improve the speed of wound repair. There was no difference between the two groups on day 0, but the gap gradually widened later. The p-values ​​in the figure are only shown due to insufficient sample size; further statistical testing with actual samples is recommended.

[0122] Based on the two cases above, it can be seen that the drug in Example 1 of this application may synergistically promote the healing process of diabetic foot ulcers through multiple mechanisms, such as effective anti-infection, reduction of inflammatory response, stimulation of granulation tissue proliferation and promotion of re-epithelialization.

Claims

1. A traditional Chinese medicine composition for treating diabetic foot ulcers, characterized in that, The effective components are obtained by decocting and extracting the raw medicinal materials.

2. The traditional Chinese medicine composition for treating diabetic foot ulcer according to claim 1, characterized in that, The prepared dosage form is an external dressing.

3. The traditional Chinese medicine composition for treating diabetic foot ulcer according to claim 1 or 2, characterized in that, The specific preparation method is as follows: the above raw materials are soaked in water, decocted, filtered, and the filtrate is collected and concentrated, and then pharmaceutically acceptable adjuvants are added to obtain the prepared dosage form.

4. The traditional Chinese medicine composition for treating diabetic foot ulcer according to claim 1 or 2, characterized in that, The water adding amount is 9-11 times the weight of the raw medicinal materials, the soaking time is 20-50 minutes, and the decocting time is 1-1.5 hours for 2-3 times.

5. The traditional Chinese medicine composition for treating diabetic foot ulcer according to claim 1 or 2, characterized in that, 10. Use of the traditional Chinese medicine composition of any one of claims 1-5 in the preparation of an external dressing for treating diabetic foot ulcers.

6. The method of preparing the traditional Chinese medicine composition for treating diabetic foot ulcer according to any one of claims 1-5, characterized in that, ​ 7. The method for preparing the traditional Chinese medicine composition for treating diabetic foot ulcers as described in claim 6, characterized in that, ​ 8. The method for preparing the traditional Chinese medicine composition for treating diabetic foot ulcers as described in claim 7, characterized in that, ​ 9. The method for preparing the traditional Chinese medicine composition for treating diabetic foot ulcers as described in claim 8, characterized in that, ​ ​

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