Medicine for repairing skin wound and preparation method thereof

Abstract

The invention relates to a medicine for repairing skin wounds and a preparation method thereof, and belongs to the technical field of biological medicines.The medicine is prepared from, by mass, 25-30 parts of bee embryo peptide, 8-10 parts of alkannin derivative, 10-15 parts of armillaria mellea polysaccharide, 8-10 parts of inonotus obliquus polysaccharide, 6-8 parts of notoginsenoside and 5-7 parts of astragaloside. The bee fetus peptide is a product which is more than 500Da and is obtained by performing step-by-step enzymolysis on bee larva freeze-dried powder through trypsin and bacillus subtilis neutral protease to obtain enzymatic hydrolysate which is less than 3kDa, performing purification through a G-50 sephadex column, performing ultrafiltration and performing freeze-drying. The alkannin derivative is obtained by reacting alkannin with succinic anhydride and glucosamine-polyethylene glycol. A special method is adopted for preparing the bee embryo peptide and alkannin derivatives, the bee embryo peptide and alkannin derivatives are matched with other components to play a synergistic role, and the effects of high safety, high skin cell proliferation promoting capacity, excellent antibacterial performance and the like are achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a medicine for repairing skin wounds and a preparation method thereof. Background Art

[0002] In daily life, skin trauma is unavoidable due to various reasons such as abrasions, burns, scalds, cuts, and chronic ulcers. Traditional treatments for skin trauma include debridement and bandaging. Early commonly used drugs such as mercurochrome and gentian violet, although they have a certain disinfecting effect, have problems such as high toxicity and easy pigmentation. With the development of medicine, antibiotics are widely used in trauma treatment, effectively reducing the risk of trauma infection, but the abuse of antibiotics has also led to new problems such as bacterial resistance. In addition, dressings such as vaseline gauze mainly serve to isolate the wound from the external environment. Although they can maintain a certain degree of moisture in the wound, they do not have the ability to promote tissue repair and the healing speed is slow.

[0003] In recent years, with the rapid development of biotechnology and materials science, research into wound repair drugs has made significant progress. Bioactive factor drugs, such as epidermal growth factor (EGF) and fibroblast growth factor (FGF), have become a research hotspot. These drugs can specifically bind to cell surface receptors, stimulating cell proliferation, differentiation, and migration, accelerating epidermal regeneration and granulation tissue formation. However, these drugs are poorly stable, easily enzymatically degraded in the wound, and their high cost limits their large-scale application.

[0004] The application of novel biomaterials in wound repair has also opened new avenues for drug development. Hydrogels, with their excellent hydrophilicity and biocompatibility, can mimic the extracellular matrix environment, supporting cell growth and migration. They can also serve as drug carriers, slowly releasing active ingredients and maintaining effective concentrations within the wound. Chitosan and its derivatives possess numerous functions, including antibacterial, hemostatic, and tissue repair. They are widely available and can be formulated into various formulations, including films and gels, for use in skin wound treatment.

[0005] However, current skin wound repair drugs still have many shortcomings. For large-area wounds or chronic, difficult-to-heal wounds, drugs with a single mechanism of action struggle to achieve the desired therapeutic effect. Furthermore, increasing the duration of drug action at the wound site and reducing immunogenicity are pressing challenges. Continuous improvement and development of novel wound repair drugs with synergistic, multifunctional properties and enhanced safety are needed to provide patients with more effective treatment outcomes. Summary of the Invention

[0006] In response to the problems of existing skin wound repair drugs, such as a single mechanism of action, short efficacy, low repair efficiency, and poor safety, the present invention provides a drug for repairing skin wounds and a preparation method thereof. Bee fetal peptide and shikonin derivatives are prepared using a special method, and then combined with purified Armillaria mellea polysaccharides and Chaga polysaccharides, and then an appropriate amount of notoginseng saponins and astragaloside IV are added. Each component is used in a certain proportion to work together, and has the effects of high safety, strong cell proliferation promotion ability, and excellent antibacterial properties. It is suitable for skin cell proliferation and repairing skin wounds. The specific technical scheme is as follows:

[0007] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = (25-30): (8-10): (10-15): (8-10): (6-8): (5-7); wherein the bee fetal peptide is prepared by stepwise enzymatic hydrolysis of bee larvae freeze-dried powder with trypsin and Bacillus subtilis neutral protease to obtain an enzymatic hydrolyzate below 3 kDa, which is then purified by G-50 dextran gel column, ultrafiltration, and freeze-drying to obtain a product above 500 Da; the shikonin derivative is prepared by reacting shikonin with succinic anhydride and glucosamine-polyethylene glycol; the Armillaria polysaccharide is prepared by re-dissolving the Armillaria polysaccharide in deionized water and then purifying it through ethanol precipitation; and the Chaga polysaccharide is prepared by re-dissolving the Chaga polysaccharide in deionized water and then purifying it through ethanol precipitation.

[0008] Among the above-mentioned drugs, the preparation method of bee fetal peptide includes: adding freeze-dried powder of bee larvae to Tris-HCl buffer with a pH of 8.0 to 8.5, adding trypsin, enzymatically hydrolyzing at 37°C to 40°C for 1.5 hours to 2 hours, inactivating the enzyme, adjusting the pH to 6 to 7, adding Bacillus subtilis neutral protease, enzymatically hydrolyzing at 50°C to 55°C for 1 hour to 1.5 hours, inactivating the enzyme, ultrafiltration through 10kDa and 3kDa ultrafiltration membranes in sequence, collecting the filtrate below 3kDa, loading it onto a G-50 dextran gel column, eluting 0 to 1.5 BV with a 0.05mol / L to 0.07mol / L NaCl aqueous solution, eluting 1.5 to 3 BV with a 0.2mol / L to 0.3mol / L NaCl aqueous solution, collecting 1.5 to 3 BV of the eluate, ultrafiltration using a 500Da ultrafiltration membrane, taking the component between 500Da and 3kDa, and freeze-drying to obtain bee fetal peptide.

[0009] In the above-mentioned preparation method of bee fetal peptide, the amount of the Tris-HCl buffer used is 6 to 8 times the mass of the lyophilized powder; the amount of the trypsin added is 0.5% to 0.8% of the mass of the lyophilized powder; and the amount of the Bacillus subtilis neutral protease added is 0.8% to 1.5% of the mass of the lyophilized powder.

[0010] In the above-mentioned preparation method of bee fetal peptide, the stirring speed of the enzymatic hydrolysis is 150 rpm to 180 rpm; and the enzyme inactivation is performed at 80° C. to 85° C. for 10 min to 15 min.

[0011] Among the above-mentioned drugs, the preparation method of the shikonin derivative comprises: dissolving shikonin in N,N-dimethylformamide (DMF), adding succinic anhydride and 4-dimethylaminopyridine (DMAP), stirring and reacting at 60°C to 65°C and 200rpm to 250rpm under nitrogen protection for 10h to 12h to obtain a reaction solution A, pouring the solution into ice water, standing to precipitate, centrifuging, taking the precipitate, washing it with anhydrous ethanol, adding it to a N,N-dimethylformamide-water mixed solvent with a pH of 5.0 to 6.0, adding glucosamine-polyethylene glycol, and then adding 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), stirring and reacting at 30°C to 35°C and 150rpm to 200rpm for 2h to 4h to obtain a reaction solution B, charging the solution into a dialysis bag, dialyzing it in flowing deionized water for 24h to 30h, and freeze-drying it to obtain the shikonin derivative.

[0012] In the above-mentioned preparation method of shikonin derivatives, the amount of N,N-dimethylformamide used is 10 to 12 times the mass of shikonin; the amount of succinic anhydride added is 1.8 to 2 times the mass of shikonin; the amount of 4-dimethylaminopyridine added is 0.03 to 0.05 times the mass of shikonin; the amount of ice water used is 3 to 5 times the volume of reaction solution A; the centrifugation is 6000 rpm to 8000 rpm for 15 to 20 minutes; and the anhydrous ethanol washing is performed 3 to 4 times.

[0013] In the above-mentioned preparation method of shikonin derivatives, the amount of the N,N-dimethylformamide-water mixed solvent is 10 to 12 times the mass of the precipitate, the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is (7 to 8): (2 to 3), and the pH of the N,N-dimethylformamide-water mixed solvent is adjusted to 5.5 to 6.0 with 0.15M to 0.2M MES buffer; the amount of glucosamine-polyethylene glycol added is 0.75 to 0.95 times the mass of the precipitate; the amount of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride added is 0.95 to 1.05 times the mass of the precipitate; the amount of N-hydroxysuccinimide added is 0.55 to 0.65 times the mass of the precipitate; and the molecular weight cutoff of the dialysis bag is 3500Da.

[0014] Among the above-mentioned drugs, Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method includes: taking Armillaria polysaccharide and adding it to 3 to 4 times the mass of deionized water, stirring and mixing at 50°C to 60°C and 500rpm to 600rpm for 20min to 30min, centrifuging at 6000rpm to 8000rpm for 10min to 15min, taking the supernatant, adding anhydrous ethanol 3 to 4 times the volume of the supernatant, standing at 4°C to 6°C for 12h to 15h to precipitate, centrifuging at 6000rpm to 8000rpm for 15min to 20min, taking the precipitate, and vacuum drying at 35°C to 40°C for 8h to 12h to obtain purified Armillaria polysaccharide.

[0015] Among the above-mentioned drugs, Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: taking Chaga polysaccharide, adding 3 to 4 times the mass of deionized water, stirring and mixing at 50°C to 60°C and 500rpm to 600rpm for 20min to 30min, centrifuging at 6000rpm to 8000rpm for 10min to 15min, taking the supernatant, adding anhydrous ethanol 3 to 4 times the volume of the supernatant, standing at 4°C to 6°C for 12h to 15h to precipitate, centrifuging at 6000rpm to 8000rpm for 15min to 20min, taking the precipitate, and vacuum drying at 35°C to 40°C for 8h to 12h to obtain purified Chaga polysaccharide.

[0016] The preparation method of the above-mentioned drug for repairing skin wounds includes the following steps: mixing bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically acceptable excipients.

[0017] The present invention provides a drug for repairing skin wounds and a preparation method thereof, which has the following beneficial effects: First, in the preparation of bee fetal peptide, the enzymatic hydrolysis process uses specific trypsin and Bacillus subtilisin neutral protease, performed at an appropriate temperature, pH, and stirring speed to ensure that the freeze-dried powder of honey bee larvae is fully hydrolyzed and produces active peptides. Ultrafiltration and purification steps are used to isolate and purify the target peptides, ensuring their activity and purity, and obtaining bee fetal peptides with a specific molecular weight range (500Da-3kDa). This process ensures the structure and activity of bee fetal peptides. The peptides obtained by specific enzymatic hydrolysis can stimulate cell proliferation, differentiation, and anti-inflammatory effects, providing a driving force for cell repair. As a key component of the drug, its active ingredients can directly act on skin wounds, promoting cell growth and repair, while also having anti-inflammatory effects, reducing the inflammatory response of the wound.

[0018] Second, in the preparation of shikonin derivatives, shikonin is chemically modified by reacting with succinic anhydride and glucosamine-polyethylene glycol. The resulting shikonin derivatives possess enhanced stability and biological activity. This modification process imparts improved water solubility and biocompatibility, enabling them to function more effectively in the body. In pharmaceuticals, they exhibit anti-inflammatory and cell migration-promoting properties, alleviating wound inflammation, promoting cell migration to the site of injury, and accelerating wound healing.

[0019] Third, Armillaria polysaccharide undergoes purification to remove impurities, resulting in a higher purity, which enhances the stability and effectiveness of its antibacterial and immunomodulatory properties. In pharmaceuticals, it possesses antibacterial and immunomodulatory properties, inhibiting bacterial growth in wounds, regulating the body's immune response, and creating a favorable environment for wound healing.

[0020] Chaga polysaccharides are also purified to remove impurities, ensuring their antibacterial and immunomodulatory effects. In pharmaceuticals, they work synergistically with Armillaria polysaccharides to exert their antibacterial and immunomodulatory functions, enhancing the drug's overall antibacterial capacity and its regulatory effects on the wound microenvironment.

[0021] 5. Notoginseng saponins in medicine can improve local blood circulation, bringing more nutrients and oxygen to the wound site, and promoting tissue repair. Astragaloside IV, together with notoginseng saponins in medicine, can improve local blood circulation, promote tissue repair, and enhance the quality and speed of wound healing.

[0022] In summary, the drug of the present invention adopts bee fetal peptide, shikonin derivative, Armillaria polysaccharide, Chaga polysaccharide, notoginseng saponin and astragaloside IV in a certain proportion for use, has low hemolysis rate, high safety, and will not cause obvious adverse effects on the blood system; has strong cell proliferation promoting ability, can effectively promote cell proliferation, provide sufficient cell source for skin wound repair, and accelerate the wound healing process. It has excellent antibacterial properties, has a good inhibitory effect on common pathogens such as Staphylococcus aureus and Escherichia coli, reduces the risk of wound infection, and creates a good environment for wound healing. Bee fetal peptide, shikonin derivative, Armillaria polysaccharide, Chaga polysaccharide, notoginseng saponin and astragaloside IV work together to play a role, are suitable for skin cell proliferation, repair skin wounds, and have good practicality. DETAILED DESCRIPTION

[0023] The present invention will be further described below with reference to specific implementation cases, but the present invention is not limited to these embodiments.

[0024] Example 1 A drug for repairing skin wounds, wherein the mass ratio of the drug components is bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 25:8:10:8:6:5.

[0025] Among them, the preparation method of bee fetal peptide includes: adding lyophilized powder of bee larvae to 6 times the mass of Tris-HCl buffer with pH 8.0, adding 0.5% trypsin by mass of lyophilized powder, enzymatically hydrolyzing at 37°C and 150 rpm for 1.5 hours, inactivating the enzyme at 80°C for 10 minutes, adjusting the pH to 6, adding 0.8% Bacillus subtilis neutral protease by mass of lyophilized powder, enzymatically hydrolyzing at 50°C and 150 rpm for 1 hour, inactivating the enzyme at 80°C for 10 minutes, ultrafiltration through 10kDa and 3kDa ultrafiltration membranes in sequence, collecting the filtrate below 3kDa, loading it onto a G-50 dextran gel column, eluting 0-1.5 BV with 0.05mol / L NaCl aqueous solution, eluting 1.5-3 BV with 0.2mol / L NaCl aqueous solution, collecting 1.5-3 BV of eluate, ultrafiltration using a 500Da ultrafiltration membrane, taking the component between 500Da and 3kDa, and freeze-drying to obtain bee fetal peptide.

[0026] The preparation method of the shikonin derivative comprises: dissolving shikonin in 10 times the mass of N,N-dimethylformamide (DMF), adding succinic anhydride (1.8 times the mass of shikonin) and 4-dimethylaminopyridine (DMAP) (0.03 times the mass of shikonin), stirring and reacting at 60°C and 200 rpm under nitrogen protection for 10 hours to obtain a reaction solution A, pouring into ice water (3 times the volume of the reaction solution A), standing to precipitate, centrifuging at 6000 rpm for 15 minutes, taking the precipitate, washing it with anhydrous ethanol three times, and adding 10 times the mass of the precipitate pH 5.0 In an N,N-dimethylformamide-water mixed solvent, wherein the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is 7:2, and the pH is adjusted to 5.5 with 0.2M MES buffer; 0.75 times the mass of the precipitate of glucosamine-polyethylene glycol, 0.95 times the mass of the precipitate of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.55 times the mass of the precipitate of N-hydroxysuccinimide (NHS) are added, and the mixture is stirred at 30°C and 150 rpm for 2 hours to obtain a reaction solution B, which is charged into a dialysis bag with a molecular weight cutoff of 3500 Da, dialyzed in flowing deionized water for 24 hours, and freeze-dried to obtain a shikonin derivative.

[0027] Among them, Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method includes: taking Armillaria polysaccharide and adding it to 3 times the mass of deionized water, stirring and mixing at 50°C and 500rpm for 20 minutes, centrifuging at 6000rpm for 10 minutes, taking the supernatant, adding anhydrous ethanol 3 times the volume of the supernatant, standing at 4°C for 12 hours to precipitate, centrifuging at 6000rpm for 15 minutes, taking the precipitate, and vacuum drying at 35°C for 8 hours to obtain purified Armillaria polysaccharide.

[0028] Among them, the Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: taking Chaga polysaccharide and adding it to 3 times the mass of deionized water, stirring and mixing at 50°C and 500rpm for 20 minutes, centrifuging at 6000rpm for 10 minutes, taking the supernatant, adding anhydrous ethanol 3 times the volume of the supernatant, standing at 4°C for 12 hours to precipitate, centrifuging at 6000rpm for 15 minutes, taking the precipitate, and vacuum drying at 35°C for 8 hours to obtain purified Chaga polysaccharide.

[0029] The preparation method of the above-mentioned drug for repairing skin wounds includes the following steps: mixing bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically acceptable excipients.

[0030] Example 2 A drug for repairing skin wounds, wherein the mass ratio of the drug components is bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 28:9:13:9:7:6.

[0031] Among them, the preparation method of bee fetal peptide includes: adding lyophilized powder of bee larvae to 7 times the mass of Tris-HCl buffer with pH 8.2, adding 0.65% trypsin by mass of lyophilized powder, enzymatically hydrolyzing at 38°C and 160rpm for 1.5h, inactivating the enzyme at 82°C for 12min, adjusting the pH to 6.5, adding 1.2% Bacillus subtilis neutral protease by mass of lyophilized powder, enzymatically hydrolyzing at 52°C and 160rpm for 1.5h, inactivating the enzyme at 82°C for 12min, ultrafiltration through 10kDa and 3kDa ultrafiltration membranes in sequence, collecting the filtrate below 3kDa, loading it onto a G-50 dextran gel column, eluting with 0.06mol / L NaCl aqueous solution for 0-1.5 BV, eluting with 0.25mol / L NaCl aqueous solution for 1.5-3 BV, and collecting 1.5-3 The eluate of BV was ultrafiltered using a 500Da ultrafiltration membrane, and the components between 500Da and 3kDa were taken and freeze-dried to obtain bee fetal peptide.

[0032] The preparation method of the shikonin derivative comprises: dissolving shikonin in 11 times the mass of N,N-dimethylformamide (DMF), adding succinic anhydride (1.9 times the mass of shikonin) and 4-dimethylaminopyridine (DMAP) (0.04 times the mass of shikonin), stirring and reacting at 62°C and 220 rpm for 11 hours under nitrogen protection to obtain a reaction solution A, pouring into ice water (4 times the volume of the reaction solution A), standing to precipitate, centrifuging at 7000 rpm for 18 minutes, taking the precipitate, washing it with anhydrous ethanol three times, and adding 11 times the mass of the precipitate, pH 5.5 In an N,N-dimethylformamide-water mixed solvent, wherein the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is 7.5:2.5, and the pH is adjusted to 5.8 with 0.15M MES buffer; 0.85 times the mass of the precipitate of glucosamine-polyethylene glycol, 1 times the mass of the precipitate of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.6 times the mass of the precipitate of N-hydroxysuccinimide (NHS) are added, and the mixture is stirred at 32°C and 180 rpm for 3 hours to obtain a reaction solution B, which is charged into a dialysis bag with a molecular weight cutoff of 3500 Da, dialyzed in flowing deionized water for 26 hours, and freeze-dried to obtain a shikonin derivative.

[0033] Among them, the Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method includes: taking Armillaria polysaccharide and adding it to 3.5 times the mass of deionized water, stirring and mixing at 55°C and 550rpm for 25 minutes, centrifuging at 7000rpm for 12 minutes, taking the supernatant, adding anhydrous ethanol 3.5 times the volume of the supernatant, standing at 5°C for 14 hours to precipitate, centrifuging at 7000rpm for 18 minutes, taking the precipitate, and vacuum drying at 38°C for 10 hours to obtain purified Armillaria polysaccharide.

[0034] Among them, the Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: taking Chaga polysaccharide and adding it to 3.5 times the mass of deionized water, stirring and mixing at 55°C and 550rpm for 25 minutes, centrifuging at 7000rpm for 12 minutes, taking the supernatant, adding anhydrous ethanol 3.5 times the volume of the supernatant, standing at 5°C for 13 hours to precipitate, centrifuging at 7000rpm for 18 minutes, taking the precipitate, and vacuum drying at 38°C for 10 hours to obtain purified Chaga polysaccharide.

[0035] The preparation method of the above-mentioned drug for repairing skin wounds includes the following steps: mixing bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically acceptable excipients.

[0036] Example 3 A drug for repairing skin wounds, wherein the mass ratio of the drug components is bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 30:10:15:10:8:7.

[0037] Among them, the preparation method of bee fetal peptide includes: adding lyophilized powder of bee larvae to 8 times the mass of Tris-HCl buffer with pH 8.5, adding 0.8% trypsin by mass of the lyophilized powder, enzymatically hydrolyzing at 40°C and 180rpm for 2h, inactivating the enzyme at 85°C for 15min, adjusting the pH to 7, adding 1.5% Bacillus subtilis neutral protease by mass of the lyophilized powder, enzymatically hydrolyzing at 55°C and 180rpm for 1.5h, inactivating the enzyme at 85°C for 15min, ultrafiltration through 10kDa and 3kDa ultrafiltration membranes in sequence, collecting the filtrate below 3kDa, loading it onto a G-50 dextran gel column, eluting 0-1.5 BV with 0.07mol / L NaCl aqueous solution, eluting 1.5-3 BV with 0.3mol / L NaCl aqueous solution, collecting 1.5-3 BV of the eluate, ultrafiltration using a 500Da ultrafiltration membrane, taking the component between 500Da and 3kDa, and freeze-drying to obtain bee fetal peptide.

[0038] The preparation method of the shikonin derivative comprises: dissolving shikonin in 12 times the mass of N,N-dimethylformamide (DMF), adding succinic anhydride (2 times the mass of shikonin) and 4-dimethylaminopyridine (DMAP) (0.05 times the mass of shikonin), stirring and reacting at 65°C and 250 rpm for 12 hours under nitrogen protection to obtain a reaction solution A, pouring into ice water (5 times the volume of the reaction solution A), standing to precipitate, centrifuging at 8000 rpm for 20 minutes, taking the precipitate, washing it with anhydrous ethanol 4 times, and adding 12 times the mass of the precipitate, pH 6.0 In an N,N-dimethylformamide-water mixed solvent, wherein the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is 8:3, and the pH is adjusted to 6.0 with 0.2M MES buffer; 0.95 times the mass of the precipitate of glucosamine-polyethylene glycol, 1.05 times the mass of the precipitate of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.65 times the mass of the precipitate of N-hydroxysuccinimide (NHS) are added, and the mixture is stirred at 35°C and 200 rpm for 4 hours to obtain a reaction solution B, which is charged into a dialysis bag with a molecular weight cutoff of 3500 Da, dialyzed in flowing deionized water for 30 hours, and freeze-dried to obtain a shikonin derivative.

[0039] Among them, Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method includes: taking Armillaria polysaccharide and adding it to 4 times the mass of deionized water, stirring and mixing at 60°C and 600rpm for 30 minutes, centrifuging at 8000rpm for 15 minutes, taking the supernatant, adding anhydrous ethanol 4 times the volume of the supernatant, standing at 6°C for 15 hours to precipitate, centrifuging at 8000rpm for 20 minutes, taking the precipitate, and vacuum drying at 40°C for 12 hours to obtain purified Armillaria polysaccharide.

[0040] Among them, the Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: taking Chaga polysaccharide and adding it to 4 times the mass of deionized water, stirring and mixing at 60°C and 600rpm for 30 minutes, centrifuging at 8000rpm for 15 minutes, taking the supernatant, adding anhydrous ethanol 4 times the volume of the supernatant, standing at 6°C for 15 hours to precipitate, centrifuging at 8000rpm for 20 minutes, taking the precipitate, and vacuum drying at 40°C for 12 hours to obtain purified Chaga polysaccharide.

[0041] The preparation method of the above-mentioned drug for repairing skin wounds includes the following steps: mixing bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically acceptable excipients.

[0042] Example 4 A drug for repairing skin wounds, wherein the mass ratio of the drug components is bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 25:10:13:10:6:7.

[0043] Among them, the preparation method of bee fetal peptide includes: adding lyophilized powder of bee larvae to 6 times the mass of Tris-HCl buffer with pH 8.5, adding 0.5% trypsin by mass of the lyophilized powder, enzymatically hydrolyzing at 40°C and 150rpm for 2h, inactivating the enzyme at 80°C for 15min, adjusting the pH to 6, adding 1.5% Bacillus subtilis neutral protease by mass of the lyophilized powder, enzymatically hydrolyzing at 50°C and 180rpm for 1h, inactivating the enzyme at 85°C for 10min, ultrafiltration through 10kDa and 3kDa ultrafiltration membranes in sequence, collecting the filtrate below 3kDa, loading it onto a G-50 dextran gel column, eluting 0-1.5 BV with 0.07mol / L NaCl aqueous solution, eluting 1.5-3 BV with 0.2mol / L NaCl aqueous solution, collecting 1.5-3 BV of the eluate, ultrafiltration using a 500Da ultrafiltration membrane, taking the component between 500Da and 3kDa, and freeze-drying to obtain bee fetal peptide.

[0044] The preparation method of the shikonin derivative comprises: dissolving shikonin in 12 times the mass of N,N-dimethylformamide (DMF), adding succinic anhydride (1.8 times the mass of shikonin) and 4-dimethylaminopyridine (DMAP) (0.05 times the mass of shikonin), stirring and reacting at 60°C and 250 rpm for 10 hours under nitrogen protection to obtain a reaction solution A, pouring into ice water (5 times the volume of the reaction solution A), standing to precipitate, centrifuging at 6000 rpm for 20 minutes, taking the precipitate, washing it with anhydrous ethanol three times, and adding 12 times the mass of the precipitate, pH 5.0 In an N,N-dimethylformamide-water mixed solvent, wherein the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is 7:3, and the pH is adjusted to 5.6 with 0.15M MES buffer; 0.78 times the mass of the precipitate of glucosamine-polyethylene glycol, 1.02 times the mass of the precipitate of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.55 times the mass of the precipitate of N-hydroxysuccinimide (NHS) are added, and the mixture is stirred at 35°C and 150 rpm for 4 hours to obtain a reaction solution B, which is loaded into a dialysis bag with a molecular weight cutoff of 3500Da, dialyzed in flowing deionized water for 24 hours, and freeze-dried to obtain a shikonin derivative.

[0045] Among them, Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method includes: taking Armillaria polysaccharide and adding it to 4 times the mass of deionized water, stirring and mixing at 50°C and 600rpm for 20 minutes, centrifuging at 8000rpm for 10 minutes, taking the supernatant, adding anhydrous ethanol 4 times the volume of the supernatant, standing at 4°C for 15 hours to precipitate, centrifuging at 6000rpm for 20 minutes, taking the precipitate, and vacuum drying at 35°C for 12 hours to obtain purified Armillaria polysaccharide.

[0046] Among them, the Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: taking Chaga polysaccharide and adding it to 3 times the mass of deionized water, stirring and mixing at 60°C and 500rpm for 30 minutes, centrifuging at 6000rpm for 15 minutes, taking the supernatant, adding anhydrous ethanol 3 times the volume of the supernatant, standing at 6°C for 12 hours to precipitate, centrifuging at 8000rpm for 15 minutes, taking the precipitate, and vacuum drying at 40°C for 8 hours to obtain purified Chaga polysaccharide.

[0047] The preparation method of the above-mentioned drug for repairing skin wounds includes the following steps: mixing bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically acceptable excipients.

[0048] In the above embodiments, the honey bee species was Apis mellifera. Trypsin was sourced from Ningxia Xiasheng Industrial Group Co., Ltd., model FDG-2281, with an enzyme activity of 200,000 U / g. Bacillus subtilis neutral protease was sourced from Nanning Pangbo Bioengineering Co., Ltd., with an enzyme activity of 200,000 U / g. G-50 dextran gel column was sourced from Shanghai Yuanye Biotechnology Co., Ltd. Shikonin was sourced from Wellman Pharmaceutical Group Co., Ltd., with a purity of 99%. N,N-dimethylformamide (DMF) was sourced from Shanghai Jingkang Bioengineering Co., Ltd. Succinic anhydride was sourced from Shandong Xinheng Chemical Co., Ltd., with a pharmaceutical grade of 99.5%. 4-Dimethylaminopyridine (DMAP) was sourced from Wuhan Lanabai Pharmaceutical Chemical Co., Ltd. MES (2-(N-morpholino)ethanesulfonic acid monohydrate) was sourced from Wuxi Tongchuang Biotechnology Co., Ltd. Glucosamine-polyethylene glycol was sourced from Xi'an Ruixi Biotechnology Co., Ltd., with a molecular weight of 4 kDa. 1-Ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) was sourced from Wuhan Xinweiye Chemical Co., Ltd. N-hydroxysuccinimide (NHS) was sourced from Shanghai Yixin Biotechnology Co., Ltd. Armillaria polysaccharide was sourced from Shaanxi Dongjiang Kangtai Health Industry Co., Ltd., with a purity of 50%. Chaga polysaccharide was sourced from Fufeng Sinuote Biotechnology Co., Ltd., with a purity of 60%. Notoginseng saponins were sourced from Zhuhai Jiayi Biotechnology Co., Ltd., with Notoginseng saponin R1 (80% pharmaceutical grade). Astragaloside IV was sourced from Shanxi Yuning Biotechnology Co., Ltd., with a pharmaceutical grade of 98%.

[0049] Comparative Example 1 Bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 15:18:10:8:6:5; other parameters and methods are the same as in Example 1.

[0050] Comparative Example 2 Bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 35:3:10:8:6:5; other parameters and methods are the same as in Example 1.

[0051] Comparative Example 3 Bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 25:8:3:15:6:5; other parameters and methods are the same as in Example 1.

[0052] Comparative Example 4 Bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = 25:8:20:3:6:5; other parameters and methods are the same as in Example 1.

[0053] Comparative Example 5 In the preparation method of bee fetal peptide, trypsin is replaced by alkaline protease; other parameters and methods are the same as in Example 1.

[0054] Comparative Example 6 In the preparation method of bee fetal peptide, Bacillus subtilisin neutral protease is replaced by bromelain; other parameters and methods are the same as in Example 1.

[0055] Comparative Example 7 In the preparation method of bee-fetoprotein, G-50 glucan gel is replaced by LH-60 glucan gel; other parameters and methods are the same as in Example 1.

[0056] Comparative Example 8 The shikonin derivative was replaced by shikonin; other parameters and methods were the same as those in Example 1.

[0057] Comparative Example 9 In the preparation method of shikonin derivatives, the precipitate after washing with anhydrous ethanol is directly dialyzed without glucosamine-polyethylene glycol modification; other parameters and methods are the same as those in Example 1.

[0058] Comparative Example 10 In the preparation method of shikonin derivatives, glucosamine-polyethylene glycol is replaced by polyethylene glycol (molecular weight 4000); other parameters and methods are the same as in Example 1.

[0059] Comparative Example 11 The Armillaria polysaccharide was not purified; other parameters and methods were the same as in Example 1.

[0060] Comparative Example 12 Chaga polysaccharide was not purified; other parameters and methods were the same as in Example 1.

[0061] Sources of the replacements in the above comparative examples: Alkaline protease (derived from Bacillus licheniformis) was sourced from Ningxia Xiasheng Industrial Group Co., Ltd., with an enzyme activity of 200,000 U / g. Bromelain was sourced from Shaanxi Zelang Biotechnology Co., Ltd., with an enzyme activity of 200,000 U / g. LH-60 dextran gel was sourced from Shanghai Yuanye Biotechnology Co., Ltd.; and polyethylene glycol (molecular weight 4000) was sourced from Xi'an Xinfengda Pharmaceutical Excipients Co., Ltd.

[0062] 1. Hemolysis rate test: Sample preparation: The drugs prepared in each example and comparative example were prepared into solutions with a concentration of 0.5 mg / mL using normal saline.

[0063] The test method involves taking fresh anticoagulated human blood, washing it three times with normal saline, centrifuging it at 2000 rpm for 5 minutes each time, discarding the supernatant, and diluting the red blood cells to 2% (v / v) with normal saline. A test tube is then filled with 2 mL of the test solution, with three replicates of each type. A positive control (equal volume of distilled water) and a negative control (equal volume of normal saline) are also established, along with a drug-containing control (drug-free, cell-free) to subtract background absorbance. 0.2 mL of the red blood cell suspension is then added to each tube, gently mixed, and incubated at 37°C in a water bath for 1 hour. The tubes are centrifuged at 2000 rpm for 5 minutes, and the supernatant is collected. The absorbance is measured at 540 nm using a spectrophotometer. Hemolysis rate (%) = (OD value of the experimental group - OD value of the drug-containing control group - OD value of the negative control group) / (OD value of the positive control group - OD value of the negative control group) × 100%.

[0064] 2. Cell proliferation promotion detection: Sample preparation: The drugs prepared in each example and comparative example were prepared into solutions with a concentration of 0.1 mg / mL using normal saline.

[0065] Detection method: CCK-8 method was used. The method includes: human epidermal keratinocytes were cultured at 3×10 4 Cells were seeded at a density of 1 cell / well in a 96-well plate, with 100 μL of cell-containing culture medium per well. After 24 hours of culture, 100 μL of the test sample was added, and five replicates were set up. A blank control (culture medium, no cells) and a negative control (cells + culture medium) were also set up, along with a drug background control (drug + culture medium, no cells) to subtract background absorbance. After 72 hours of culture, the culture medium was discarded, and 100 μL of fresh culture medium containing 10% CCK-8 was added to each well. Incubation continued for another 4 hours. Absorbance was measured at 450 nm using a microplate reader. Cell proliferation rate (%) = (OD value of the experimental group - OD value of the drug background control group - OD value of the blank group) / (OD value of the negative control group - OD value of the blank group) × 100%.

[0066] 3. Antibacterial performance testing: Sample preparation: The drugs prepared in each example and comparative example were prepared into solutions with a concentration of 5 mg / mL using normal saline.

[0067] Detection method: The inhibition zone method is used. The method includes: selecting activated Staphylococcus aureus and Escherichia coli as test bacteria. Adjust the concentration of the bacterial solution to 10 8CFU / mL. Sterilize agar medium and cool it to 45°C. Add 0.1 mL of bacterial solution per 100 mL, mix thoroughly, and pour onto a plate to create a bacterial plate. Use a sterile hole punch to punch a 6 mm hole in the plate and add 50 μL of the sample solution into the hole. Three replicates of the drug test group were prepared. The plates were incubated in a 37°C incubator for 24 hours, and the diameter of the inhibition zone was measured. Additionally, a solvent control group (normal saline) and an antibiotic positive control group (10 μg / mL gentamicin for Staphylococcus aureus and 10 μg / mL ampicillin for Escherichia coli) were established.

[0068] Table 1 Test results (parallel sample interval values)

[0069] Note: In the antibacterial activity test, the diameter of the inhibition zone against Staphylococcus aureus in the gentamicin positive control group was 19.4 mm; the diameter of the inhibition zone against Escherichia coli in the ampicillin positive control group was 18.2 mm; the diameter of the inhibition zone against Staphylococcus aureus in the solvent control group (normal saline) was 6.8 mm; and the diameter of the inhibition zone against Escherichia coli in the solvent control group (normal saline) was 6.6 mm. Staphylococcus aureus DSM45902 was obtained from Ningbo Testo Biotechnology Co., Ltd. Escherichia coli 0157:H7 was obtained from Shanghai Xuanya Biotechnology Co., Ltd.

[0070] It can be seen from the above results that in each embodiment, the proportion of the drug ingredients fluctuates within a reasonable range, and the overall performance is good. Bee fetal peptide, shikonin derivatives, Armillaria polysaccharide, Chaga polysaccharide, notoginseng saponin and astragaloside IV act synergistically with each other. Bee fetal peptide can stimulate cell proliferation, differentiation and anti-inflammation, providing power for cell repair; shikonin derivatives have anti-inflammatory and cell migration promoting effects; Armillaria polysaccharide and Chaga polysaccharide have antibacterial and immunomodulatory functions, creating a good environment for wound healing; notoginseng saponin and astragaloside IV can improve local blood circulation and promote tissue repair. The reasonable combination of ingredient ratios enables the drug to maintain a low level in terms of hemolysis rate, while performing excellently in promoting cell proliferation and antibacterial properties, and with the optimization of the ingredient ratios, various aspects of performance are improved to a certain extent.

[0071] In Comparative Example 1, the proportion of shikonin derivatives was relatively high, while the proportion of bee fetal peptide was low, disrupting the synergistic balance between the components. Excessive shikonin derivatives can cause cellular irritation, leading to increased hemolysis. Furthermore, the insufficient bee fetal peptide content weakened the cell proliferation-promoting ability, decreased cell proliferation rate, and reduced antibacterial properties due to the disruption of the synergistic effect of the components.

[0072] In Comparative Example 2, the proportion of bee fetoprotein was too high, while the proportion of shikonin derivatives was too low. Excessive bee fetoprotein levels may cause excessive cell stress responses, leading to increased hemolysis rates. Insufficient shikonin derivatives, on the other hand, weakened their anti-inflammatory and cell migration-promoting effects, resulting in a lower cell proliferation rate and compromised antibacterial properties.

[0073] In Comparative Example 3, the proportion of Armillaria polysaccharide was too low, while the proportion of Chaga polysaccharide was too high. The reduction in Armillaria polysaccharide partially compromised its antibacterial and immunomodulatory abilities. While Chaga polysaccharide also exhibited antibacterial effects, the imbalanced ratio resulted in a poor overall synergistic antibacterial effect, with reduced inhibition zone diameters for Staphylococcus aureus and Escherichia coli. Furthermore, the imbalanced immune regulation compromised the normal cell growth environment, increasing the hemolysis rate.

[0074] In Comparative Example 4, in contrast to Comparative Example 3, the proportion of Armillaria polysaccharide was too high, while the proportion of Chaga polysaccharide was too low. Excessive Armillaria polysaccharide can lead to excessive accumulation of some components, affecting drug permeability and cellular absorption of the active ingredient. Furthermore, the insufficient Chaga polysaccharide content prevented its full antibacterial and immunomodulatory effects, resulting in a reduced cell proliferation rate, a loss of antibacterial synergy, and a reduced inhibition zone diameter.

[0075] In Comparative Example 5, alkaline protease was used in place of trypsin in the preparation of bee fetal peptide. Different proteases have different action sites and enzymatic hydrolysis characteristics. Substituting alkaline protease for trypsin altered the enzymatic hydrolysis process of lyophilized honey bee larvae powder, preventing the production of active peptides with the same structure and function as the bee fetal peptide in Example 1. This resulted in reduced activity of the bee fetal peptide, which in turn affected the overall performance of the drug.

[0076] In Comparative Example 6, the preparation method for bee-fetal peptide (BPP) replaced Bacillus subtilisin with bromelain. Bromelain and Bacillus subtilisin differ in their enzymatic specificity, and this substitution altered the preparation process of the bee-fetal peptide, resulting in changes in the structure and activity of the resulting bee-fetal peptide. With reduced activity, the bee-fetal peptide was unable to effectively promote cell proliferation and exert other functions.

[0077] In Comparative Example 7, in the preparation method of bee-fetoprotein peptide, G-50 dextran gel was replaced with LH-60 dextran gel. The two gels have different pore sizes and separation properties. Substituting LH-60 dextran gel for G-50 dextran gel altered the separation efficiency of bee-fetoprotein peptide, preventing the effective isolation of active peptides within the appropriate molecular weight range. This affected the purity and activity of bee-fetoprotein peptide, and consequently, reduced its hemolysis rate, cell proliferation promotion, and antibacterial properties.

[0078] In Comparative Example 8, shikonin was used as a shikonin derivative instead. Shikonin derivatives are prepared through a series of chemical modifications and have better stability and bioactivity than shikonin. Directly using shikonin as a replacement lacks the advantages of chemical modification, such as anti-inflammatory, cell migration-promoting, and reduced ability to synergize with other ingredients.

[0079] In Comparative Example 9, the precipitate from the shikonin derivative was directly dialyzed after washing with anhydrous ethanol without undergoing glucosamine-polyethylene glycol modification. While glucosamine-polyethylene glycol modification can impart improved water solubility and biocompatibility to the shikonin derivative, lack of modification results in poor in vivo distribution and efficacy of the shikonin derivative, impacting the overall performance of the drug.

[0080] In Comparative Example 10, in the preparation method of a shikonin derivative, glucosamine-PEG was replaced with polyethylene glycol (molecular weight 4000). Glucosamine-PEG has a unique structure similar to glucosamine, enabling it to bind to cell surface receptors. However, the substitution with polyethylene glycol results in different binding modes and effects between polyethylene glycol and shikonin than with glucosamine-PEG, resulting in altered properties of the shikonin derivative.

[0081] In Comparative Example 11, Armillaria polysaccharide was not purified. Unpurified Armillaria polysaccharide contains a large amount of impurities, which can affect the stability, biological activity, and safety of the drug. The presence of impurities interferes with the normal interaction between the drug and cells.

[0082] In Comparative Example 12, Chaga polysaccharide was not purified. Similar to the unpurified Armillaria polysaccharide, unpurified Chaga polysaccharide contains impurities that affect the drug's performance. Impurities can interfere with the synergistic effects of the drug's components, reducing its antibacterial and cell repair abilities.

Claims

1. A drug for repairing skin wounds, characterized in that: The mass ratio of the drug components is bee fetal peptide: shikonin derivative: Armillaria polysaccharide: Chaga polysaccharide: notoginseng saponin: astragaloside IV = (25-30): (8-10): (10-15): (8-10): (6-8): (5-7); the bee fetal peptide is prepared by stepwise enzymatic hydrolysis of bee larvae freeze-dried powder with trypsin and Bacillus subtilis neutral protease to obtain an enzymatic hydrolyzate below 3 kDa, which is then purified by G-50 dextran gel column, ultrafiltration, and freeze-drying to obtain a product above 500 Da; the shikonin derivative is prepared by the reaction of shikonin with succinic anhydride and glucosamine-polyethylene glycol; the Armillaria polysaccharide is prepared by re-dissolving Armillaria polysaccharide in deionized water and then purifying it through ethanol precipitation; the Chaga polysaccharide is prepared by re-dissolving Chaga polysaccharide in deionized water and then purifying it through ethanol precipitation.

2. A drug for repairing skin wounds according to claim 1, characterized in that: The preparation method of the bee fetal peptide comprises the following steps: adding lyophilized powder of honey bee larvae to a Tris-HCl buffer solution with a pH of 8.0 to 8.5, adding trypsin, performing enzymatic hydrolysis at 37° C. to 40° C. for 1.5 to 2 hours, inactivating the enzyme, adjusting the pH to 6 to 7, adding Bacillus subtilis neutral protease, performing enzymatic hydrolysis at 50° C. to 55° C. for 1 to 1.5 hours, inactivating the enzyme, performing ultrafiltration through 10 kDa and 3 kDa ultrafiltration membranes in sequence, collecting the filtrate below 3 kDa, loading the filtrate onto a G-50 dextran gel column, eluting 0 to 1.5 BV with a 0.05 mol / L to 0.07 mol / L NaCl aqueous solution, eluting 1.5 to 3 BV with a 0.2 mol / L to 0.3 mol / L NaCl aqueous solution, collecting 1.5 to 3 BV of the eluate, ultrafiltration using a 500 Da ultrafiltration membrane, taking the component between 500 Da and 3 kDa, and freeze-drying to obtain the bee fetal peptide.

3. A drug for repairing skin wounds according to claim 2, characterized in that: The amount of the Tris-HCl buffer used is 6 to 8 times the mass of the lyophilized powder; the amount of trypsin added is 0.5% to 0.8% of the mass of the lyophilized powder; and the amount of the Bacillus subtilis neutral protease added is 0.8% to 1.5% of the mass of the lyophilized powder.

4. The drug for repairing skin wounds according to claim 2, characterized in that: The stirring speed of the enzymatic hydrolysis is 150 rpm to 180 rpm; the enzyme inactivation is carried out at 80° C. to 85° C. for 10 min to 15 min.

5. The drug for repairing skin wounds according to claim 1, characterized in that: The preparation method of the shikonin derivative comprises the following steps: dissolving shikonin in N,N-dimethylformamide, adding succinic anhydride and 4-dimethylaminopyridine, stirring and reacting at 60° C. to 65° C. and 200 rpm to 250 rpm under nitrogen protection for 10 to 12 hours to obtain a reaction solution A, pouring the solution into ice water, standing to separate out a precipitate, centrifuging, taking the precipitate, washing the precipitate with anhydrous ethanol, adding the solution into a N,N-dimethylformamide-water mixed solvent with a pH of 5.0 to 6.0, adding glucosamine-polyethylene glycol, and then adding 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride and N-hydroxysuccinimide, stirring and reacting at 30° C. to 35° C. and 150 rpm to 200 rpm for 2 to 4 hours to obtain a reaction solution B, charging the solution into a dialysis bag, dialyzing the solution in flowing deionized water for 24 to 30 hours, and freeze-drying the solution to obtain the shikonin derivative.

6. The drug for repairing skin wounds according to claim 5, characterized in that: The amount of N,N-dimethylformamide used is 10 to 12 times the mass of lithospermum; the amount of succinic anhydride added is 1.8 to 2 times the mass of lithospermum; the amount of 4-dimethylaminopyridine added is 0.03 to 0.05 times the mass of lithospermum; the amount of ice water used is 3 to 5 times the volume of reaction solution A; the centrifugation is performed at 6000 to 8000 rpm for 15 to 20 minutes; and the anhydrous ethanol washing is performed 3 to 4 times.

7. The drug for repairing skin wounds according to claim 5, characterized in that: The amount of the N,N-dimethylformamide-water mixed solvent is 10 to 12 times the mass of the precipitate, the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is (7 to 8): (2 to 3), and the pH of the N,N-dimethylformamide-water mixed solvent is adjusted to 5.5 to 6.0 with 0.15M to 0.2M MES buffer; the amount of glucosamine-polyethylene glycol added is 0.75 to 0.95 times the mass of the precipitate; the amount of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride added is 0.95 to 1.05 times the mass of the precipitate; the amount of N-hydroxysuccinimide added is 0.55 to 0.65 times the mass of the precipitate; and the molecular weight cutoff of the dialysis bag is 3500Da.

8. The drug for repairing skin wounds according to claim 1, characterized in that: The Armillaria polysaccharide is purified Armillaria polysaccharide, and the purification method comprises: adding Armillaria polysaccharide to deionized water 3 to 4 times its mass, stirring and mixing at 50°C to 60°C and 500rpm to 600rpm for 20min to 30min, centrifuging at 6000rpm to 8000rpm for 10min to 15min, taking the supernatant, adding anhydrous ethanol 3 to 4 times the volume of the supernatant, standing at 4°C to 6°C for 12h to 15h to precipitate, centrifuging at 6000rpm to 8000rpm for 15min to 20min, taking the precipitate, and vacuum drying at 35°C to 40°C for 8h to 12h to obtain the purified Armillaria polysaccharide.

9. The drug for repairing skin wounds according to claim 1, characterized in that: The Chaga polysaccharide is purified Chaga polysaccharide, and the purification method includes: adding Chaga polysaccharide to deionized water 3 to 4 times its mass, stirring and mixing at 50° C. to 60° C. and 500 rpm to 600 rpm for 20 to 30 minutes, centrifuging at 6000 rpm to 8000 rpm for 10 to 15 minutes, taking the supernatant, adding anhydrous ethanol 3 to 4 times the volume of the supernatant, standing at 4° C. to 6° C. for 12 to 15 hours to precipitate, centrifuging at 6000 rpm to 8000 rpm for 15 to 20 minutes, taking the precipitate, and vacuum drying at 35° C. to 40° C. for 8 to 12 hours to obtain the purified Chaga polysaccharide.

10. The method for preparing a drug for repairing skin wounds according to claim 1, characterized in that: The method comprises the following steps: mixing bee fetal peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, notoginseng saponin and astragaloside IV according to mass ratio to obtain a pharmaceutical composition; and preparing a dressing preparation with the pharmaceutical composition and pharmaceutically usable excipients.

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