Medicament for repairing skin wounds and its preparation method

Abstract

The application relates to a medicine for repairing skin wounds and a preparation method thereof, and belongs to the technical field of biological medicines. The mass ratio of the components of the medicine is (25-30):(8-10):(10-15):(8-10):(6-8):(5-7) of bee larva peptide, shikonin derivative, armillaria mellea polysaccharide, phellinus igniarius polysaccharide, notoginseng saponin and astragaloside. The bee larva peptide is obtained by sequentially subjecting freeze-dried powder of bee larvae to trypsin and neutral protease of bacillus subtilis, purifying the 3kDa or smaller enzymatic solution through a G-50 dextran gel column, ultrafiltrating and freeze-drying to obtain a product with a molecular weight of 500Da or more. The shikonin derivative is obtained by reacting shikonin with succinic anhydride and glucosamine-polyethylene glycol. The bee larva peptide and the shikonin derivative are prepared by a special method, and are combined with other components to synergistically play a role, and have the effects of high safety, strong skin cell proliferation promoting capacity, excellent antibacterial performance and the like.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a drug for repairing skin wounds and its preparation method. Background Technology

[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 medications, such as mercurochrome and gentian violet, while having some disinfecting effect, have problems such as high toxicity and a tendency to cause pigmentation. With the development of medicine, antibiotics are widely used in wound treatment, effectively reducing the risk of wound infection. However, the overuse of antibiotics has also led to new problems such as bacterial resistance. In addition, dressings such as petroleum jelly gauze mainly serve to isolate the wound from the external environment. Although they can maintain a certain level of wound moisture, they do not have the ability to promote tissue repair, resulting in a slower healing rate.

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

[0004] The application of novel biomaterials in wound repair has also brought new directions to drug development. Hydrogels possess excellent hydrophilicity and biocompatibility, enabling them to mimic the extracellular matrix environment, supporting cell growth and migration. They can also serve as drug carriers, slowly releasing active ingredients and maintaining effective drug concentrations at the wound site. Chitosan and its derivatives possess multiple functions, including antibacterial, hemostatic, and tissue repair-promoting properties, and are widely available, allowing for the formulation of films, gels, and other dosage forms for the treatment of skin wounds.

[0005] However, current skin wound repair medications still have many shortcomings. For large-area wounds or chronic, difficult-to-heal wounds, drugs with a single mechanism of action are insufficient to achieve ideal therapeutic effects. Furthermore, improving the duration of drug action at the wound site and reducing immunogenicity are also pressing issues that need to be addressed. There is a need to continuously improve and develop novel wound repair drugs with synergistic effects and high safety profiles to provide patients with more effective treatment outcomes. Summary of the Invention

[0006] To address the problems of existing skin wound repair drugs, such as single mechanism of action, short duration of action, low repair efficiency, and poor safety, this invention provides a drug for repairing skin wounds and its preparation method. A special method is used to prepare bee placenta peptide and shikonin derivatives, which are then combined with purified Armillaria mellea polysaccharide and Chaga polysaccharide, and further supplemented with appropriate proportions of notoginsenosides and astragaloside A. The components are used in a specific ratio to work synergistically, exhibiting high safety, strong cell proliferation promotion ability, and excellent antibacterial properties. It is suitable for skin cell proliferation and skin wound repair. The specific technical solution is as follows:

[0007] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee placenta peptide: shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponin: Astragaloside A = (25-30): (8-10): (10-15): (8-10): (6-8): (5-7); wherein, bee placenta peptide is obtained by stepwise enzymatic hydrolysis of freeze-dried bee larvae powder by trypsin and Bacillus subtilis neutral protease to obtain an enzymatic hydrolysate with a value below 3kDa, purified by G-50 dextran gel column, ultrafiltered, and freeze-dried to obtain a product with a value above 500Da; shikonin derivative is obtained by reacting shikonin with succinic anhydride and glucosamine-polyethylene glycol; the Armillaria mellea polysaccharide is obtained by reconstituted Armillaria mellea polysaccharide with deionized water and purified by ethanol precipitation; the Chaga polysaccharide is obtained by reconstituted Chaga polysaccharide with deionized water and purified by ethanol precipitation.

[0008] The preparation method of the above-mentioned drug, namely bee placenta peptide, includes: adding freeze-dried powder of bee larvae to Tris-HCl buffer at pH 8.0-8.5, adding trypsin, enzymatically hydrolyzing at 37℃-40℃ for 1.5-2 hours, inactivating the enzyme, adjusting the pH to 6-7, adding Bacillus subtilis neutral protease, enzymatically hydrolyzing at 50℃-55℃ for 1-1.5 hours, inactivating the enzyme, and then ultrafiltration through 10kDa and 3kDa ultrafiltration membranes sequentially. The filtrate below 3kDa is collected and loaded onto a G-50 dextran gel column. 0-1.5 BV is eluted with 0.05mol / L-0.07mol / L NaCl aqueous solution, and 1.5-3 BV is eluted with 0.2mol / L-0.3mol / L NaCl aqueous solution. The eluent of 1.5-3 BV is collected and ultrafiltered using a 500Da ultrafiltration membrane. The fraction between 500Da and 3kDa is taken, freeze-dried, and bee placenta peptide is obtained.

[0009] In the above-mentioned method for preparing bee placenta peptide, the amount of Tris-HCl buffer 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 Bacillus subtilis neutral protease added is 0.8% to 1.5% of the mass of the lyophilized powder.

[0010] In the above-mentioned method for preparing bee placenta peptide, the stirring speed for enzymatic hydrolysis is 150 rpm to 180 rpm; and the enzyme inactivation is carried out at 80℃ to 85℃ for 10 min to 15 min.

[0011] The preparation method of the above-mentioned drug, namely, shikonin derivative, includes: dissolving shikonin in N,N-dimethylformamide (DMF), adding succinic anhydride and 4-dimethylaminopyridine (DMAP), and stirring under nitrogen protection at 60℃~65℃ and 200rpm~250rpm for 10h~12h to obtain reaction solution A. The solution is poured into ice water, allowed to stand to precipitate, centrifuged, and the precipitate is washed with anhydrous ethanol. Then, it is added to a pH 5.0~6.0 N,N-dimethylformamide-water mixed solvent, followed by glucosamine-polyethylene glycol, and then 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The solution is stirred at 30℃~35℃ and 150rpm~200rpm for 2h~4h to obtain reaction solution B. This solution is placed in a dialysis bag and dialyzed in flowing deionized water for 24h~30h. Finally, it is freeze-dried to obtain the shikonin derivative.

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

[0013] In the above-mentioned method for preparing the shikonin derivative, 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 (7-8):(2-3); the pH of the N,N-dimethylformamide-water mixed solvent is adjusted to 5.5-6.0 using 0.15M-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 3500 Da.

[0014] In the above-mentioned drugs, the *Armillaria mellea* polysaccharide is purified *Armillaria mellea* polysaccharide. The purification method includes: adding *Armillaria mellea* polysaccharide to 3 to 4 times its mass of deionized water, stirring and mixing at 50℃ to 60℃ 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 3 to 4 times the volume of the supernatant of anhydrous ethanol, letting it stand at 4℃ to 6℃ 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℃ to 40℃ for 8 to 12 hours to obtain purified *Armillaria mellea* polysaccharide.

[0015] Among the above-mentioned drugs, the chaga polysaccharide is the purified chaga polysaccharide. The purification method includes: adding 3 to 4 times the mass of the chaga polysaccharide to deionized water, stirring and mixing at 50℃ to 60℃ 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 3 to 4 times the volume of the supernatant to anhydrous ethanol, letting it stand at 4℃ to 6℃ 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℃ to 40℃ for 8 to 12 hours to obtain the purified chaga polysaccharide.

[0016] The preparation method of the above-mentioned drug for repairing skin trauma includes the following steps: mixing bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside A in a mass ratio to obtain a drug composition; and preparing a dressing preparation by combining the drug composition with pharmaceutically available excipients.

[0017] The present invention provides a drug for repairing skin trauma and a method for preparing the same, with the following beneficial effects:

[0018] In the preparation of bee placenta peptides, the enzymatic hydrolysis process utilizes specific trypsin and Bacillus subtilis neutral protease, carried out under suitable temperature, pH, and stirring speed to ensure that the freeze-dried bee larvae powder is fully hydrolyzed, producing active peptides. Ultrafiltration and purification steps are then used to separate and purify the target peptides, ensuring their activity and purity, yielding bee placenta peptides with a specific molecular weight range (500 Da-3 kDa). This process ensures the structure and activity of the bee placenta peptides; the peptides obtained through specific enzymatic hydrolysis can stimulate cell proliferation, differentiation, and anti-inflammation, providing impetus for cell repair. As an important component of the drug, its activity can directly act on the skin wound site, promoting cell growth and repair, while also possessing anti-inflammatory effects, reducing wound inflammation.

[0019] II. In the preparation of shikonin derivatives, shikonin undergoes chemical modification through reactions with succinic anhydride, glucosamine-polyethylene glycol, etc., resulting in shikonin derivatives with improved stability and bioactivity. The modification process imparts better water solubility and biocompatibility, enabling them to exert their effects more effectively in vivo. In pharmaceuticals, they exhibit anti-inflammatory and cell migration-promoting effects, reducing wound inflammation, promoting cell migration to the wound site, and accelerating wound healing.

[0020] III. Through purification steps, impurities are removed to obtain high-purity *Armillaria mellea* polysaccharides, improving the stability and effectiveness of their antibacterial and immunomodulatory functions. In pharmaceuticals: they possess antibacterial and immunomodulatory functions, inhibiting bacterial growth at wound sites, regulating the body's immune response, and creating a favorable environment for wound healing.

[0021] IV. Chaga polysaccharides also undergo purification to remove impurities, ensuring their antibacterial and immunomodulatory effects. In pharmaceuticals: they act synergistically with Armillaria mellea polysaccharides to jointly exert antibacterial and immunomodulatory functions, enhancing the overall antibacterial ability of the drug and its regulatory effect on the wound microenvironment.

[0022] Fifth, in medications, notoginseng saponins can improve local blood circulation, bringing more nutrients and oxygen to the wound site and promoting tissue repair. Astragaloside A, in medications, works together with notoginseng saponins to improve local blood circulation, promote tissue repair, and enhance the quality and speed of wound healing.

[0023] In summary, the drug of this invention utilizes a specific ratio of bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponins, and astragaloside A. It exhibits low hemolysis, high safety, and no significant adverse effects on the blood system. It possesses strong cell proliferation-promoting capabilities, effectively promoting cell proliferation and providing a sufficient cell source for skin wound repair, thus accelerating the wound healing process. It also demonstrates excellent antibacterial properties, exhibiting good inhibitory effects against common pathogenic bacteria such as Staphylococcus aureus and Escherichia coli, reducing the risk of wound infection and creating a favorable environment for wound healing. The synergistic effects of bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponins, and astragaloside A make it suitable for skin cell proliferation and skin wound repair, demonstrating good practicality. Detailed Implementation

[0024] The present invention will be further described below with reference to specific implementation examples, but the present invention is not limited to these embodiments.

[0025] Example 1

[0026] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee placenta peptide: shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponins: Astragaloside A = 25:8:10:8:6:5.

[0027] The preparation method of bee placenta peptide includes: adding lyophilized bee larvae powder to 6 times its mass of pH 8.0 Tris-HCl buffer, adding 0.5% (by weight of the lyophilized powder) of trypsin, enzymatically hydrolyzing at 37℃ and 150 rpm for 1.5 h, inactivating the enzyme at 80℃ for 10 min, adjusting the pH to 6, adding 0.8% (by weight of the lyophilized powder) of Bacillus subtilis neutral protease, enzymatically hydrolyzing at 50℃ and 150 rpm for 1 h, inactivating the enzyme at 80℃ for 10 min, and then ultrafiltration through 10 kDa and 3 kDa ultrafiltration membranes. The filtrate below 3 kDa is collected and loaded onto a G-50 dextran gel column. The fractions from 0 to 1.5 BV are eluted with 0.05 mol / L NaCl aqueous solution, and the fractions from 1.5 to 3 BV are eluted with 0.2 mol / L NaCl aqueous solution. The eluent from 1.5 to 3 BV is collected and ultrafiltered using a 500 Da ultrafiltration membrane. The fractions between 500 Da and 3 kDa are collected, lyophilized, and the bee placenta peptide is obtained.

[0028] The preparation method of the shikonin derivative includes: dissolving shikonin in 10 times its mass of N,N-dimethylformamide (DMF), adding 1.8 times its mass of shikonin succinic anhydride and 0.03 times its mass of shikonin 4-dimethylaminopyridine (DMAP), and stirring the mixture at 60℃ and 200 rpm for 10 h under nitrogen protection to obtain reaction solution A. The solution is then poured into ice water with a volume three times that of reaction solution A, allowed to stand to precipitate, centrifuged at 6000 rpm for 15 min, and the precipitate is collected. After washing three times with anhydrous ethanol, 10 times its mass of pH 5.0 solution is added. In a mixed solvent of N,N-dimethylformamide and water, where the volume ratio of N,N-dimethylformamide to water is 7:2, the pH is adjusted to 5.5 with 0.2M MES buffer. 0.75 times the precipitate mass of glucosamine-polyethylene glycol is added, followed by 0.95 times the precipitate mass of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.55 times the precipitate mass of N-hydroxysuccinimide (NHS). The mixture is stirred at 30°C and 150 rpm for 2 hours to obtain reaction solution B. This solution is then placed in a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in flowing deionized water for 24 hours. After freeze-drying, the shikonin derivative is obtained.

[0029] Among them, the *Armillaria mellea* polysaccharide is the purified *Armillaria mellea* polysaccharide. The purification method includes: adding *Armillaria mellea* polysaccharide to 3 times its mass of deionized water, stirring and mixing at 500 rpm for 20 min at 50℃, centrifuging at 6000 rpm for 10 min, taking the supernatant, adding 3 times the volume of anhydrous ethanol to the supernatant, letting it stand at 4℃ for 12 h to precipitate, centrifuging at 6000 rpm for 15 min, taking the precipitate, and vacuum drying at 35℃ for 8 h to obtain the purified *Armillaria mellea* polysaccharide.

[0030] Among them, the chaga polysaccharide is the purified chaga polysaccharide. The purification method includes: adding 3 times the mass of chaga polysaccharide to deionized water, stirring and mixing at 500 rpm for 20 min at 50 ℃, centrifuging at 6000 rpm for 10 min, taking the supernatant, adding 3 times the volume of anhydrous ethanol to the supernatant, letting it stand at 4 ℃ for 12 h to precipitate, centrifuging at 6000 rpm for 15 min, taking the precipitate, and vacuum drying at 35 ℃ for 8 h to obtain the purified chaga polysaccharide.

[0031] The preparation method of the above-mentioned drug for repairing skin trauma includes the following steps: mixing bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside A in a mass ratio to obtain a drug composition; and preparing a dressing preparation by combining the drug composition with pharmaceutically available excipients.

[0032] Example 2

[0033] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee placenta peptide: shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponin: Astragaloside A = 28:9:13:9:7:6.

[0034] The preparation method of bee placenta peptide includes: adding 7 times the mass of freeze-dried bee larvae powder to a pH 8.2 Tris-HCl buffer, adding 0.65% (by weight of the freeze-dried powder) of trypsin, hydrolyzing at 38℃ and 160 rpm for 1.5 h, inactivating the enzyme at 82℃ for 12 min, adjusting the pH to 6.5, adding 1.2% (by weight of the freeze-dried powder) of Bacillus subtilis neutral protease, hydrolyzing at 52℃ and 160 rpm for 1.5 h, inactivating the enzyme at 82℃ for 12 min, and then ultrafiltration through 10 kDa and 3 kDa ultrafiltration membranes. The filtrate below 3 kDa is collected and loaded onto a G-50 dextran gel column. Elute 0–1.5 BV with 0.06 mol / L NaCl aqueous solution, and 1.5–3 BV with 0.25 mol / L NaCl aqueous solution. Collect the 1.5–3 BV filtrate. The eluent of BV was ultrafiltered using a 500Da ultrafiltration membrane, and the fraction between 500Da and 3kDa was collected, then lyophilized to obtain bee placenta peptide.

[0035] The preparation method of the shikonin derivative includes: dissolving shikonin in 11 times its mass of N,N-dimethylformamide (DMF), adding 1.9 times its mass of shikonin succinic anhydride and 0.04 times its mass of shikonin 4-dimethylaminopyridine (DMAP), and stirring the mixture at 62℃ and 220 rpm for 11 h under nitrogen protection to obtain reaction solution A. The solution is then poured into 4 times its volume of ice water, allowed to stand to precipitate, centrifuged at 7000 rpm for 18 min, and the precipitate is collected. After washing three times with anhydrous ethanol, 11 times its mass of pH 5.5 is added. In a mixed solvent of N,N-dimethylformamide and water, the volume ratio of N,N-dimethylformamide to water was 7.5:2.5. The pH was adjusted to 5.8 with 0.15M MES buffer. 0.85 times the precipitate mass of glucosamine-polyethylene glycol was added, followed by 1 times the precipitate mass of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.6 times the precipitate mass of N-hydroxysuccinimide (NHS). The mixture was stirred at 32°C and 180 rpm for 3 hours to obtain reaction solution B. This solution was then placed in a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in flowing deionized water for 26 hours. After freeze-drying, the shikonin derivative was obtained.

[0036] Among them, the *Armillaria mellea* polysaccharide is the purified *Armillaria mellea* polysaccharide. The purification method includes: adding *Armillaria mellea* polysaccharide to 3.5 times its mass of deionized water, stirring and mixing at 55℃ and 550 rpm for 25 min, centrifuging at 7000 rpm for 12 min, taking the supernatant, adding 3.5 times the volume of the supernatant of anhydrous ethanol, letting it stand at 5℃ for 14 h to precipitate, centrifuging at 7000 rpm for 18 min, taking the precipitate, and vacuum drying at 38℃ for 10 h to obtain the purified *Armillaria mellea* polysaccharide.

[0037] Among them, the chaga polysaccharide is the purified chaga polysaccharide. The purification method includes: adding 3.5 times the mass of chaga polysaccharide to deionized water, stirring and mixing at 55℃ and 550rpm for 25min, centrifuging at 7000rpm for 12min, taking the supernatant, adding 3.5 times the volume of anhydrous ethanol to the supernatant, letting it stand at 5℃ for 13h to precipitate, centrifuging at 7000rpm for 18min, taking the precipitate, and vacuum drying at 38℃ for 10h to obtain the purified chaga polysaccharide.

[0038] The preparation method of the above-mentioned drug for repairing skin trauma includes the following steps: mixing bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside A in a mass ratio to obtain a drug composition; and preparing a dressing preparation by combining the drug composition with pharmaceutically available excipients.

[0039] Example 3

[0040] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee placenta peptide: shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponin: Astragaloside A = 30:10:15:10:8:7.

[0041] The preparation method of bee placenta peptide includes: adding lyophilized bee larvae powder to 8 times its mass of pH 8.5 Tris-HCl buffer, adding 0.8% (by weight of the lyophilized powder) of trypsin, hydrolyzing at 40℃ and 180 rpm for 2 h, inactivating the enzyme at 85℃ for 15 min, adjusting the pH to 7, adding 1.5% (by weight of the lyophilized powder) of Bacillus subtilis neutral protease, hydrolyzing at 55℃ and 180 rpm for 1.5 h, inactivating the enzyme at 85℃ for 15 min, and then passing the solution through 10 kDa and 3 kDa ultrafiltration membranes sequentially. The filtrate below 3 kDa is collected and loaded onto a G-50 dextran gel column. The fractions from 0 to 1.5 BV are eluted with 0.07 mol / L NaCl aqueous solution, and the fractions from 1.5 to 3 BV are eluted with 0.3 mol / L NaCl aqueous solution. The eluent from 1.5 to 3 BV is collected and ultrafiltered using a 500 Da ultrafiltration membrane. The fractions between 500 Da and 3 kDa are collected, lyophilized, and the bee placenta peptide is obtained.

[0042] The preparation method of the shikonin derivative includes: dissolving shikonin in 12 times its mass of N,N-dimethylformamide (DMF), adding 2 times its mass of shikonin succinic anhydride and 0.05 times its mass of shikonin 4-dimethylaminopyridine (DMAP), and stirring the mixture at 65℃ and 250 rpm for 12 h under nitrogen protection to obtain reaction solution A. The solution is then poured into 5 times its volume of ice water, allowed to stand to precipitate, centrifuged at 8000 rpm for 20 min, and the precipitate is collected. After washing four times with anhydrous ethanol, 12 times its mass of pH 6.0 solution is added. In a mixed solvent of N,N-dimethylformamide and water, where the volume ratio of N,N-dimethylformamide to water is 8:3, the pH is adjusted to 6.0 with 0.2M MES buffer. 0.95 times the precipitate mass of glucosamine-polyethylene glycol is added, followed by 1.05 times the precipitate mass of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.65 times the precipitate mass of N-hydroxysuccinimide (NHS). The mixture is stirred at 35°C and 200 rpm for 4 hours to obtain reaction solution B. This solution is then placed in a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in flowing deionized water for 30 hours. After freeze-drying, the shikonin derivative is obtained.

[0043] Among them, the *Armillaria mellea* polysaccharide is the purified *Armillaria mellea* polysaccharide. The purification method includes: adding *Armillaria mellea* polysaccharide to 4 times its mass of deionized water, stirring and mixing at 60℃ and 600 rpm for 30 min, centrifuging at 8000 rpm for 15 min, taking the supernatant, adding 4 times the volume of anhydrous ethanol to the supernatant, letting it stand at 6℃ for 15 h to precipitate, centrifuging at 8000 rpm for 20 min, taking the precipitate, and vacuum drying at 40℃ for 12 h to obtain the purified *Armillaria mellea* polysaccharide.

[0044] Among them, the chaga polysaccharide is the purified chaga polysaccharide. The purification method includes: adding chaga polysaccharide to 4 times its mass of deionized water, stirring and mixing at 60℃ and 600rpm for 30min, centrifuging at 8000rpm for 15min, taking the supernatant, adding 4 times the volume of anhydrous ethanol to the supernatant, letting it stand at 6℃ for 15h to precipitate, centrifuging at 8000rpm for 20min, taking the precipitate, and vacuum drying at 40℃ for 12h to obtain the purified chaga polysaccharide.

[0045] The preparation method of the above-mentioned drug for repairing skin trauma includes the following steps: mixing bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside A in a mass ratio to obtain a drug composition; and preparing a dressing preparation by combining the drug composition with pharmaceutically available excipients.

[0046] Example 4

[0047] A drug for repairing skin wounds, wherein the mass ratio of the drug components is: bee placenta peptide: shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponin: Astragaloside A = 25:10:13:10:6:7.

[0048] The preparation method of bee placenta peptide includes: adding lyophilized bee larvae powder to 6 times its mass of pH 8.5 Tris-HCl buffer, adding 0.5% (by weight of the lyophilized powder) of trypsin, enzymatically hydrolyzing at 40℃ and 150 rpm for 2 h, inactivating the enzyme at 80℃ for 15 min, adjusting the pH to 6, adding 1.5% (by weight of the lyophilized powder) of Bacillus subtilis neutral protease, enzymatically hydrolyzing at 50℃ and 180 rpm for 1 h, inactivating the enzyme at 85℃ for 10 min, and then ultrafiltration through 10 kDa and 3 kDa ultrafiltration membranes. The filtrate below 3 kDa is collected and loaded onto a G-50 dextran gel column. The fractions from 0 to 1.5 BV are eluted with 0.07 mol / L NaCl aqueous solution, and the fractions from 1.5 to 3 BV are eluted with 0.2 mol / L NaCl aqueous solution. The eluent from 1.5 to 3 BV is collected and ultrafiltered using a 500 Da ultrafiltration membrane. The fractions between 500 Da and 3 kDa are collected, lyophilized, and the bee placenta peptide is obtained.

[0049] The preparation method of the shikonin derivative includes: dissolving shikonin in 12 times its mass of N,N-dimethylformamide (DMF), adding 1.8 times its mass of shikonin succinic anhydride and 0.05 times its mass of shikonin 4-dimethylaminopyridine (DMAP), and reacting under nitrogen protection at 60℃ and 250 rpm for 10 h with stirring to obtain reaction solution A. The solution is then poured into 5 times its volume of ice water, allowed to stand to precipitate, centrifuged at 6000 rpm for 20 min, and the precipitate is collected. After washing three times with anhydrous ethanol, 12 times its mass of pH 5.0 solution is added. In a mixed solvent of N,N-dimethylformamide and water, where the volume ratio of N,N-dimethylformamide to water is 7:3, the pH is adjusted to 5.6 with 0.15M MES buffer. 0.78 times the precipitate mass of glucosamine-polyethylene glycol is added, followed by 1.02 times the precipitate mass of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) and 0.55 times the precipitate mass of N-hydroxysuccinimide (NHS). The mixture is stirred at 35°C and 150 rpm for 4 hours to obtain reaction solution B. This solution is then placed in a dialysis bag with a molecular weight cutoff of 3500 Da and dialyzed in flowing deionized water for 24 hours. After freeze-drying, the shikonin derivative is obtained.

[0050] Among them, the *Armillaria mellea* polysaccharide is the purified *Armillaria mellea* polysaccharide. The purification method includes: adding *Armillaria mellea* polysaccharide to 4 times its mass of deionized water, stirring and mixing at 50℃ and 600 rpm for 20 min, centrifuging at 8000 rpm for 10 min, taking the supernatant, adding 4 times the volume of anhydrous ethanol to the supernatant, letting it stand at 4℃ for 15 h to precipitate, centrifuging at 6000 rpm for 20 min, taking the precipitate, and vacuum drying at 35℃ for 12 h to obtain the purified *Armillaria mellea* polysaccharide.

[0051] Among them, the chaga polysaccharide is the purified chaga polysaccharide. The purification method includes: adding 3 times the mass of chaga polysaccharide to deionized water, stirring and mixing at 60℃ and 500rpm for 30min, centrifuging at 6000rpm for 15min, taking the supernatant, adding 3 times the volume of anhydrous ethanol to the supernatant, letting it stand at 6℃ for 12h to precipitate, centrifuging at 8000rpm for 15min, taking the precipitate, and vacuum drying at 40℃ for 8h to obtain the purified chaga polysaccharide.

[0052] The preparation method of the above-mentioned drug for repairing skin trauma includes the following steps: mixing bee placenta peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponin and astragaloside A in a mass ratio to obtain a drug composition; and preparing a dressing preparation by combining the drug composition with pharmaceutically available excipients.

[0053] In the above embodiments, the bee species used was the honeybee (Apis cerana). 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., pharmaceutical grade 99.5%. 4-Dimethylaminopyridine (DMAP) was sourced from Wuhan Lanabai Pharmaceutical Chemical Co., Ltd. MES was 2-(N-morpholine)ethanesulfonic acid monohydrate, 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) is sourced from Shanghai Yixin Biotechnology Co., Ltd. Armillaria mellea polysaccharide is sourced from Shaanxi Dongjiang Kangtai Health Industry Co., Ltd., with a purity of 50%. Chaga polysaccharide is sourced from Fufeng Sinote Biotechnology Co., Ltd., with a purity of 60%. Panax notoginseng saponins are sourced from Zhuhai Jiayi Biotechnology Co., Ltd., specifically Panax notoginseng saponin R1, with a pharmaceutical grade of 80%. Astragaloside A is sourced from Shanxi Yuning Biotechnology Co., Ltd., with a pharmaceutical grade of 98%.

[0054] Comparative Example 1

[0055] Bee placenta peptide: Shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponins: Astragaloside A = 15:18:10:8:6:5; other parameters and methods are the same as in Example 1.

[0056] Comparative Example 2

[0057] Bee placenta peptide: Shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponins: Astragaloside A = 35:3:10:8:6:5; other parameters and methods are the same as in Example 1.

[0058] Comparative Example 3

[0059] Bee placenta peptide: Shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponins: Astragaloside A = 25:8:3:15:6:5; other parameters and methods are the same as in Example 1.

[0060] Comparative Example 4

[0061] Bee placenta peptide: Shikonin derivative: Armillaria mellea polysaccharide: Chaga polysaccharide: Panax notoginseng saponins: Astragaloside A = 25:8:20:3:6:5; other parameters and methods are the same as in Example 1.

[0062] Comparative Example 5

[0063] In the preparation method of bee placenta peptide, trypsin is replaced with alkaline protease; other parameters and methods are the same as in Example 1.

[0064] Comparative Example 6

[0065] In the preparation method of bee placenta peptide, Bacillus subtilis neutral protease is replaced with bromelain; other parameters and methods are the same as in Example 1.

[0066] Comparative Example 7

[0067] In the preparation method of bee placenta peptide, G-50 dextran gel is replaced with LH-60 dextran gel; other parameters and methods are the same as in Example 1.

[0068] Comparative Example 8

[0069] The shikonin derivative was replaced with shikonin; other parameters and methods were the same as in Example 1.

[0070] Comparative Example 9

[0071] 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 in Example 1.

[0072] Comparative Example 10

[0073] In the preparation method of shikonin derivative, glucosamine-polyethylene glycol is replaced by polyethylene glycol (molecular weight 4000); other parameters and methods are the same as in Example 1.

[0074] Comparative Example 11

[0075] The Armillaria mellea polysaccharide was not purified; other parameters and methods were the same as in Example 1.

[0076] Comparative Example 12

[0077] The chaga polysaccharide was not purified; other parameters and methods were the same as in Example 1.

[0078] The sources of the substitutes in the above comparative examples are as follows: Alkaline protease (derived from Bacillus licheniformis) was from Ningxia Xiasheng Industrial Group Co., Ltd., with an enzyme activity of 200,000 U / g. Bromelain was from Shaanxi Zelang Biotechnology Co., Ltd., with an enzyme activity of 200,000 U / g. LH-60 dextran gel was from Shanghai Yuanye Biotechnology Co., Ltd. Polyethylene glycol (molecular weight 4000) was from Xi'an Xinfengda Pharmaceutical Excipients Co., Ltd.

[0079] I. Hemolysis rate test:

[0080] Sample preparation: The drugs prepared in each example and comparative example were respectively prepared into solutions with physiological saline with a concentration of 0.5 mg / mL.

[0081] The detection method included: taking fresh anticoagulated human blood, washing it three times with physiological saline, centrifuging at 2000 rpm for 5 min each time, discarding the supernatant, and diluting the red blood cells with physiological saline to 2% (v / v). Adding 2 mL of sample solution to each test tube, with three parallel samples per sample; simultaneously setting up a positive control group (equal volume of distilled water) and a negative control group (equal volume of physiological saline), and a drug background control group (drug, cell-free) to subtract the absorbance background value; then adding 0.2 mL of red blood cell suspension to each tube, gently mixing, and incubating at 37℃ for 1 h. Centrifuging at 2000 rpm for 5 min, collecting the supernatant, and measuring the absorbance at 540 nm using a spectrophotometer. Hemolysis rate (%) = (OD value of experimental group - OD value of drug background control group - OD value of negative control group) / (OD value of positive control group - OD value of negative control group) × 100%.

[0082] II. Cell proliferation promotion detection:

[0083] Sample preparation: The drugs prepared in each example and comparative example were respectively prepared into solutions with physiological saline with a concentration of 0.1 mg / mL.

[0084] Detection method: CCK-8 assay was used. The method included: human epidermal keratinocytes were sampled at a concentration of 3 × 10⁻⁶ cells / mL. 4 Cells were seeded at a density of 100 μL / well in 96-well plates, with 100 μL of cell-containing medium per well. After 24 h of culture, 100 μL of the sample was added, with 5 replicates. A blank control group (medium, cell-free) and a negative control group (cells + medium) were also included. A drug background control group (drug + medium, cell-free) was also set up to subtract background absorbance values. After 72 h of culture, the medium was discarded, and 100 μL of fresh medium containing 10% CCK-8 was added to each well, followed by incubation for another 4 h. Absorbance was measured at 450 nm using a microplate reader. Cell proliferation rate (%) = (Experimental group OD value - Drug background control group OD value - Blank group OD value) / (Negative control group OD value - Blank group OD value) × 100%.

[0085] III. Antibacterial performance testing:

[0086] Sample preparation: The drugs prepared in each example and comparative example were respectively prepared into solutions with physiological saline with a concentration of 5 mg / mL.

[0087] Detection method: The inhibition zone method was used. The method included selecting activated Staphylococcus aureus and Escherichia coli as test strains. The bacterial concentration was adjusted to 10... 8CFU / mL. Take sterilized agar medium, cool it to 45℃, add 0.1mL of bacterial solution per 100mL, mix well, and pour into a Petri dish to prepare bacterial plates. Use a sterile punch to make 6mm holes in the plate, and add 50μL of sample solution to each hole; the sample drug test group consists of 3 parallel samples. Incubate the plates in a 37℃ incubator for 24h and measure the diameter of the inhibition zone. In addition, a solvent control group (physiological saline) and an antibiotic positive control group (10μg / mL gentamicin for Staphylococcus aureus, and 10μg / mL ampicillin for Escherichia coli) are set up.

[0088] Table 1. Test results (parallel sample interval values)

[0089]

[0090] Note: In the antibacterial performance testing experiment, the inhibition zone diameter of the gentamicin positive control group against Staphylococcus aureus was 19.4 mm; the inhibition zone diameter of the ampicillin positive control group against Escherichia coli was 18.2 mm; the inhibition zone diameter of the solvent control group (physiological saline) against Staphylococcus aureus was 6.8 mm; and the inhibition zone diameter of the solvent control group (physiological saline) against Escherichia coli was 6.6 mm. Staphylococcus aureus was obtained from Ningbo Taisto Biotechnology Co., Ltd., Staphylococcus aureus DSM45902. Escherichia coli was obtained from Shanghai Xuanya Biotechnology Co., Ltd., Escherichia coli O157:H7.

[0091] The results above show that the proportions of the drug components fluctuated within a reasonable range in each embodiment, exhibiting good overall performance. Bee venom peptide, shikonin derivative, Armillaria mellea polysaccharide, Chaga polysaccharide, Panax notoginseng saponins, and astragaloside A act synergistically. Bee venom peptide can stimulate cell proliferation, differentiation, and anti-inflammation, providing impetus for cell repair; shikonin derivative has anti-inflammatory and cell migration-promoting effects; Armillaria mellea polysaccharide and Chaga polysaccharide have antibacterial and immunomodulatory functions, creating a favorable environment for wound healing; Panax notoginseng saponins and astragaloside A can improve local blood circulation and promote tissue repair. The reasonable combination of component proportions keeps the drug at a low level in terms of hemolysis rate, while exhibiting excellent cell proliferation promotion and antibacterial properties. Furthermore, with the optimization of component proportions, the performance in all aspects is improved to a certain extent.

[0092] In Comparative Example 1, the proportion of shikonin derivatives was relatively high, while the proportion of bee placenta peptides was low, disrupting the synergistic balance among the components. Excessive shikonin derivatives can irritate cells, leading to increased hemolysis. Simultaneously, due to insufficient bee placenta peptide content, cell proliferation promotion capacity was weakened, cell proliferation rate decreased, and antibacterial properties were also reduced due to the disruption of the synergistic effect of the components.

[0093] In Comparative Example 2, the proportion of bee placenta peptides was too high, while the proportion of shikonin derivatives was too low. Excessive bee placenta peptides may cause excessive cellular stress, leading to increased hemolysis. Insufficient shikonin derivatives weaken its anti-inflammatory and cell migration-promoting effects, resulting in lower cell proliferation rate and impaired antibacterial properties.

[0094] In Comparative Example 3, the proportion of *Armillaria mellea* polysaccharide was too low, while the proportion of *Chaga mushroom* polysaccharide was too high. The reduction in *Armillaria mellea* polysaccharide resulted in a partial loss of its antibacterial and immunomodulatory capabilities. While *Chaga mushroom* polysaccharide also has antibacterial activity, the imbalance in its proportion led to poor overall synergistic antibacterial effect, and reduced the diameter of the inhibition zones for *Staphylococcus aureus* and *Escherichia coli*. Simultaneously, the imbalance in immune regulation also affected the normal cell growth environment, increasing the hemolysis rate.

[0095] In Comparative Example 4, unlike Comparative Example 3, the proportion of *Armillaria mellea* polysaccharide was too high, while the proportion of *Chaga mushroom* polysaccharide was too low. Excessive *Armillaria mellea* polysaccharide can lead to the excessive accumulation of some components, affecting drug permeability and cellular absorption of the active ingredients. Simultaneously, insufficient *Chaga mushroom* polysaccharide content prevents the full realization of its antibacterial and immunomodulatory effects, reduces cell proliferation rate, disrupts the synergistic antibacterial effect, and decreases the diameter of the inhibition zone.

[0096] In Comparative Example 5, the preparation method of bee placenta peptide involved replacing trypsin with an alkaline protease. Different proteases have different action sites and enzymatic hydrolysis characteristics. Replacing trypsin with alkaline protease alters the enzymatic hydrolysis process of freeze-dried bee larvae powder, preventing the production of active peptides with the same structure and function as the bee placenta peptide in Example 1. This results in reduced activity of the bee placenta peptide, thus affecting the overall performance of the drug.

[0097] In Comparative Example 6, bromelain was used instead of Bacillus subtilis neutral protease in the preparation method of bee placenta peptides. Bromelain and Bacillus subtilis neutral protease have different enzymatic hydrolytic specificities. This substitution altered the preparation process of bee placenta peptides, resulting in changes to the structure and activity of the final bee placenta peptides. The reduced activity of bee placenta peptides prevented them from effectively promoting cell proliferation and performing other functions.

[0098] In Comparative Example 7, the preparation method of bee placenta peptide involved replacing the G-50 dextran gel with an LH-60 dextran gel. The two gels have different pore sizes and separation characteristics. Replacing the G-50 dextran gel with the LH-60 dextran gel altered the separation effect on bee placenta peptide, failing to effectively separate active peptides within the appropriate molecular weight range, thus affecting the purity and activity of the bee placenta peptide. This consequently led to a decline in the drug's performance in terms of hemolysis rate, cell proliferation promotion, and antibacterial properties.

[0099] In Comparative Example 8, shikonin derivatives were used as a substitute for shikonin. Shikonin derivatives are prepared through a series of chemical modifications and exhibit better stability and bioactivity compared to shikonin. Directly replacing shikonin lacks the advantages brought by chemical modification, such as reduced anti-inflammatory, cell migration-promoting, and synergistic effects with other components.

[0100] In Comparative Example 9, the precipitate after washing with anhydrous ethanol was directly dialyzed without glucosamine-polyethylene glycol modification. Glucosamine-polyethylene glycol modification can impart better water solubility and biocompatibility to the shikonin derivative. Without modification, the distribution and efficacy of the shikonin derivative in vivo are poor, affecting the overall drug performance.

[0101] In Comparative Example 10, in the preparation method of the shikonin derivative, polyethylene glycol (molecular weight 4000) was used instead of glucosamine polyethylene glycol. Glucosamine polyethylene glycol has the special structure of glucosamine and can bind to cell surface receptors. After substitution with polyethylene glycol, the binding mode and effect of polyethylene glycol with shikonin are different from those of glucosamine-polyethylene glycol, resulting in changes in the properties of the shikonin derivative.

[0102] In Comparative Example 11, the Armillaria mellea polysaccharide was not purified. Unpurified Armillaria mellea polysaccharide contains numerous impurities, which can affect the stability, bioactivity, and safety of the drug. The presence of these impurities interferes with the normal interaction between the drug and cells.

[0103] In Comparative Example 12, the Chaga polysaccharide was not purified. Similar to the unpurified Armillaria mellea polysaccharide, the unpurified Chaga polysaccharide contains impurities, which affect the drug's performance. Impurities can interfere with the synergistic effects between drug components, reducing the drug's antibacterial and cell-repair-promoting abilities.

Claims

1. A medicament for repairing skin wounds, characterized by, The component quality ratio of the medicine is as follows: bee larva peptide: shikonin derivative: armillaria mellea polysaccharide: white birch fungus polysaccharide: notoginseng saponin: astragalus glycoside = (25-30) : (8-10) : (10-15) : (8-10) : (6-8) : (5-7); the bee larva peptide is obtained by sequentially subjecting freeze-dried powder of bee larvae to trypsin and subtilisin neutral protease enzymolysis to obtain an enzymolysis liquid with a molecular weight of less than 3 kDa, purifying the enzymolysis liquid through a G-50 dextran gel column, ultrafiltration and freeze-drying to obtain a product with a molecular weight of more than 500 Da; the shikonin derivative is obtained by reacting shikonin with succinic anhydride and glucosamine-polyethylene glycol; the armillaria mellea polysaccharide is obtained by dissolving armillaria mellea polysaccharide in deionized water and purifying the solution through ethanol precipitation; and the white birch fungus polysaccharide is obtained by dissolving white birch fungus polysaccharide in deionized water and purifying the solution through ethanol precipitation. The preparation method of the shikonin derivative comprises the following steps: dissolving shikonin in N, N-dimethylformamide, adding succinic anhydride and 4-dimethylamino pyridine, stirring at 60-65 DEG C and 200-250 rpm under nitrogen protection for 10-12 hours to obtain reaction liquid A, pouring the reaction liquid A into ice water, standing to precipitate, centrifuging, taking the precipitate, washing the precipitate with anhydrous ethanol, adding the precipitate into N, N-dimethylformamide-water mixed solvent with a pH of 5.0-6.0, adding glucosamine-polyethylene glycol, adding 1-ethyl-3-dimethylamino propyl carbodiimide hydrochloride and N-hydroxysuccinimide, stirring at 30-35 DEG C and 150-200 rpm for 2-4 hours to obtain reaction liquid B, loading the reaction liquid B into a dialysis bag, dialyzing in flowing deionized water for 24-30 hours, and freeze-drying to obtain the shikonin derivative.

2. The medicament for repairing skin wounds according to claim 1, wherein The preparation method of the bee larva peptide comprises the following steps: adding freeze-dried powder of bee larvae into Tris-HCl buffer solution with a pH of 8.0-8.5, adding trypsin, carrying out enzymolysis at 37-40 DEG C for 1.5-2 hours, inactivating the enzyme, adjusting the pH to 6-7, adding subtilisin neutral protease, carrying out enzymolysis at 50-55 DEG C for 1-1.5 hours, inactivating the enzyme, sequentially subjecting the solution to ultrafiltration through 10 kDa and 3 kDa ultrafiltration membranes, collecting the filtrate with a molecular weight of less than 3 kDa, loading the filtrate into a G-50 dextran gel column, eluting with 0.05-0.07 mol / L NaCl aqueous solution for 0-1.5 BV, eluting with 0.2-0.3 mol / L NaCl aqueous solution for 1.5-3 BV, collecting the eluate for 1.5-3 BV, subjecting the eluate to ultrafiltration through a 500 Da ultrafiltration membrane, taking the component with a molecular weight of 500-3,000 Da, and freeze-drying to obtain the bee larva peptide.

3. The medicament for repairing skin wounds according to claim 2, wherein The Tris-HCl buffer solution is used in an amount of 6-8 times the mass of the freeze-dried powder; the trypsin is added in an amount of 0.5-0.8% of the mass of the freeze-dried powder; and the subtilisin neutral protease is added in an amount of 0.8-1.5% of the mass of the freeze-dried powder.

4. The medicament for repairing skin wounds according to claim 2, wherein The stirring speed of the enzymolysis is 150-180 rpm; and the inactivation of the enzyme is carried out at 80-85 DEG C for 10-15 minutes.

5. The medicament for repairing skin wounds according to claim 1, wherein In the preparation method of the shikonin derivative, the amount of N,N-dimethylformamide is 10-12 times the mass of shikonin; the amount of succinic anhydride is 1.8-2 times the mass of shikonin; the amount of 4-dimethylamino pyridine is 0.03-0.05 times the mass of shikonin; the amount of ice water is 3-5 times the volume of reaction liquid A; the centrifugation is at 6000-8000 rpm for 15-20 min; and the anhydrous ethanol is washed 3-4 times.

6. The medicament for repairing skin wounds according to claim 1, wherein In the preparation method of the shikonin derivative, the amount of N,N-dimethylformamide-water mixed solvent is 10-12 times the mass of the precipitate, the volume ratio of N,N-dimethylformamide to water in the N,N-dimethylformamide-water mixed solvent is (7-8):(2-3), the N,N-dimethylformamide-water mixed solvent is adjusted to pH 5.5-6.0 with 0.15-0.2 M MES buffer; the amount of glucosamine-polyethylene glycol is 0.75-0.95 times the mass of the precipitate; the amount of 1-ethyl-3-dimethylaminopropyl carbodiimide hydrochloride is 0.95-1.05 times the mass of the precipitate; the amount of N-hydroxysuccinimide is 0.55-0.65 times the mass of the precipitate; and the molecular weight cut-off of the dialysis bag is 3500 Da.

7. The medicament for repairing skin wounds according to claim 1, wherein The armillaria mellea polysaccharide is purified armillaria mellea polysaccharide, and the purification method comprises the following steps: adding 3-4 times the mass of deionized water to the armillaria mellea polysaccharide, stirring and mixing at 50-60°C and 500-600 rpm for 20-30 min, centrifuging at 6000-8000 rpm for 10-15 min, adding 3-4 times the volume of anhydrous ethanol to the supernatant, precipitating at 4-6°C for 12-15 h, centrifuging at 6000-8000 rpm for 15-20 min, taking the precipitate, and vacuum drying at 35-40°C for 8-12 h to obtain the purified armillaria mellea polysaccharide.

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

9. The method of claim 1, wherein the pharmaceutical for repairing skin wounds is prepared by, The method comprises the following steps: mixing bee larval peptide, shikonin derivative, armillaria mellea polysaccharide, white birch mushroom polysaccharide, panax notoginseng saponin, and astragaloside B in a mass ratio to obtain a pharmaceutical composition; and formulating the pharmaceutical composition into a dressing preparation with a pharmaceutically acceptable excipient.

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