Pharmaceutical composition for wound repair as well as preparation method and application of pharmaceutical composition

Abstract

The invention provides a pharmaceutical composition for wound repair as well as a preparation method and application thereof, and belongs to the technical field of medical health care. The medicine composition for repairing the wound surface is prepared from the following components in percentage by weight: 4.0 percent to 6 percent of PVPK30 (Polyvinyl Pyrrolidone K30), 1.0 percent to 3.0 percent of P188, 0.05 percent to 0.2 percent of benzoic acid preservative, 3.0 percent to 5.0 percent of glycerol, 0.5 percent to 1.0 percent of rhizoma bletillae gum and 3.0 percent to 6.0 percent of cockroach extract. The preparation method comprises the following steps: adding a proper amount of sterilized water at 45-65 DEG C into the bletilla striata gum to completely dissolve the bletilla striata gum to obtain a solution 1; adding a proper amount of sterilized water into PVPK30 for dissolving, and uniformly stirring to obtain a solution 2; adding P188 into the solution 2, and uniformly stirring to obtain a solution 3; adding a benzoic acid preservative into the solution 3, and uniformly stirring to obtain a solution 4; after the temperature of the solution 1 is cooled to room temperature, adding the solution 4 into the solution 1, and uniformly stirring to obtain a solution 5; and adding the cockroach extract and glycerol into the solution 5, adding the sterilized water, fixing the volume, uniformly stirring, and filling to obtain the cockroach and rhizoma bletillae medicine composition. The wound repairing effect is remarkable, and the method is simple.

Description

Technical Field

[0001] The present invention relates to the technical field of medicine and health care, and in particular to a drug combination for wound repairing, and a preparation method and application thereof. Background Art

[0002] As the first barrier of the human body, the skin plays an important role in sensing external stimuli, regulating body temperature, and protecting the human body from external harm. At the same time, it is also very vulnerable to injury. Acute and chronic skin wounds caused by various injury factors are very common. The healing of wounds is often accompanied by the formation of scar tissue and severe loss of skin appendages such as hair, sweat glands, and sebaceous glands. Skin wound repair is divided into four continuous and coordinated stages, namely hemostasis, inflammation, proliferation, and remodeling. However, the wound healing process generally cannot proceed perfectly and orderly. At any stage, various factors such as excessive inflammation, accidental infection caused by injury or large-area skin defects, and the influence of diabetes may lead to abnormal skin wound repair. The continued existence of wounds not only reduces the quality of life of patients, but also leads to an increase in medical expenses, which brings a heavy burden to patients and their families.

[0003] Traditional wound dressings, such as cotton and gauze, can only wrap and protect wounds, but cannot provide significant anti-infection capabilities, nor can they maintain a moist environment. Moreover, adhesion can cause additional damage when replacing or removing them. Therefore, developing a drug and its preparation that can quickly promote skin wound repair, is easy to use, has good biocompatibility, and is easy to remove has always been a research topic for scientists.

[0004] Spray film is a new dosage form for external use that combines the advantages of sprays and films. After administration, it can form a layer of drug film on the wound surface, which has a good mechanical barrier protection effect on the wound surface. At the same time, it has the advantages of quantitative output, convenient administration, good compliance, high drug bioavailability, and targeted and controlled-release drug delivery.

[0005] Cockroach extract contains amino acids, nucleotides, small molecule peptides, phospholipids, mucosamine and other active ingredients, which are important active substances that promote angiogenesis and granulation tissue growth. They can promote granulation tissue growth and angiogenesis, and have the effect of repairing ulcer wounds and healing wounds quickly. At the same time, while effectively relieving pain symptoms, it can effectively activate the immune active cells of the wound, thereby achieving the purpose of antibacterial and anti-inflammatory. Bletilla striata is a commonly used hemostatic and moisturizing medicinal material in the folk. The folks make powder from the dried tubers of Bletilla striata as a medicine for wound repair. The main component of Bletilla striata is Bletilla striata glue, which is a natural macromolecular polysaccharide component (BSP). Studies have shown that Bletilla striata glue has good moisturizing, antibacterial, anti-inflammatory, hemostatic and other pharmacological activities. It is also a good medicinal polymer film-forming material with good cross-linking properties. Therefore, Bletilla striata glue has the dual characteristics of medicinal and pharmaceutical excipients.

[0006] The invention strictly examines the preparation process through single factor and star point design-response surface optimization method, takes wound repair activity and film-forming property as inspection indexes, optimizes the dosage of cockroach extract and white schizonepeta glue in the prescription process and the preparation process of the composite spray agent, obtains the optimal ratio of cockroach extract and white schizonepeta glue in the composite spray agent and the optimal preparation process of the spray agent, characterizes the composite spray agent through infrared spectroscopy, and finally obtains the cockroach+white schizonepeta glue composite wound repair spray agent with good antibacterial, anti-inflammatory, moisturizing and excellent wound repair effects. Summary of the invention

[0007] The technical problem to be solved by the present invention is to address the deficiencies of the above-mentioned prior art and to propose a drug combination for wound repair and a preparation method and application thereof.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] A drug combination for wound repair, comprising the following components in weight percentage: 4.0%-6% of PVPK30, 1.0%-3.0% of P188, 0.05%-0.2% of benzoic acid preservatives, 3.0%-5.0% of glycerol, 0.5%-1.0% of white schizonepeta gum, and 3.0%-6.0% of cockroach extract.

[0010] The cockroach extract comprises 11 kinds of amino acids, 9 kinds of alkaloid components, 6 kinds of fatty acid components, 5 kinds of nucleoside components, 4 kinds of peptide components, 4 kinds of phospholipid components and 4 kinds of sugar components.

[0011] Preferably, the amino acids are C7H 13 NO3、C9H 11 NO3, C 12 H 23 NO7, C5H9NO2, C5H 10 N2O3, C6H9N3O2, C6H 14 N4O2、C9H 11 NO4, C 11 H 12 N2O3、C 10 H7NO3, C5H9NO4, the alkaloid components are C8H9NO, C 25 H 47 NO4, C5H4N4O3, C 11 H 12 N2, C8H10N4O2, C5H 15 NO4P、C9H 17 NO4, C6H6N2O, C7H 15 NO3, the fatty acid components are11 H 12 O4、C 22 H 43 NO, C 18 H 34 O2, C8H8O2, C 13 H 24 O4, C7H6O4, the nucleoside components are C4H4N2O2, C 11 H 15 N5O3S、C 11 H 15 N5O5、C 12 H 13 ClN4、C9H 13 N2O9P, the peptide components are C 10 H 20 N2O4、C9H 18 N2O3、C 14 H 24 N4O6、C7H 16 N4O2, the phospholipid components are C 44 H 80 NO8P、C 26 H 52 NO7P, C3H9O6P, C8H 20 NO6P, the sugars are C 17 H 22 O 10 、C2H8NO4P、C 12 H 24 O 11 , C 18 H 32 O 16 .

[0012] Preferably, the cockroach extract also includes C6H6O2, C 15 H 10 O4、C8H 14 O2S2、C7H6O5、C 37 H 70 O5, C3H9N.

[0013] Preferably, the preparation method of the Bletilla striata glue is as follows:

[0014] Place fresh tubers of Bletilla striata in a grinder, add deionized water at a solid-liquid ratio of 1:4-1:5, grind into slurry, heat the slurry at 55-70°C for 1.5-2.0 hours, centrifuge and filter, collect the filtrate, extract repeatedly 2-3 times, place the filtrate at 60-70°C under reduced pressure and concentrate to 30% of the original volume, add ethanol and adjust the ethanol concentration to 80%-85%, stand at 0-5°C overnight, centrifuge for 15 minutes, remove the supernatant, collect the precipitate, freeze-dry, and obtain Bletilla striata glue.

[0015] Preferably, the centrifugal speed is 3000-4000 rpm.

[0016] Preferably, the preparation method of the cockroach extract is as follows:

[0017] The crushed cockroach medicinal material is placed in an extraction tank, 80%-95% ethanol is added at a material-liquid ratio of 1:4-5, the extraction is carried out under stirring and reflux at 65°C for 2.5 hours, filtered through a 100-200 mesh screen, the filtrate is collected, the extraction is repeated 2-3 times, the obtained filtrate is concentrated under reduced pressure at 65°C, the negative pressure value is 0.02-0.05MPa and the reduced pressure is concentrated to 10%-15% of the original volume, defatted oil with a volume 2-3 times that of the concentrated liquid is added to extract and defat the concentrated liquid, the defatted liquid is filtered through a 3000-4000 mesh filter cloth, the filtrate is concentrated under reduced pressure to an extract state with a specific gravity of 1.20-1.35, and the cockroach extract is obtained, and the yield of the extract is about 5%-7% of the medicinal material.

[0018] Another technical solution of the present invention is a method for preparing a drug combination for wound repair, which specifically comprises the following steps:

[0019] S1. Add an appropriate amount of 45-65°C sterilized water to the Bletilla striata gum to completely dissolve it to obtain solution 1, and cool it;

[0020] S2, dissolve PVPK30 in an appropriate amount of sterile water and stir evenly to obtain solution 2;

[0021] S3, adding P188 to the solution 2 obtained in step S2, stirring evenly to obtain solution 3;

[0022] S4, adding a benzoic acid preservative to the solution 3 obtained in step S3, and stirring to obtain a solution 4;

[0023] S5, after the temperature of the solution 1 obtained in step S1 is cooled to room temperature, the solution 4 obtained in step S4 is added to the solution 1 and stirred evenly to obtain a solution 5;

[0024] S6, adding the cockroach extract and glycerol to the solution 5 obtained in step S5, adding sterile water to make up to volume and stirring evenly, and filling to obtain the cockroach alveoli drug combination.

[0025] Preferably, the dosage form of the drug combination is a composite spray film, and the composite spray film is packed into a spray bottle.

[0026] Preferably, the molecular weight of the white schizonepeta gum in step S1 is 3.5×10 4 -8.7×10 4 Da, the moisture absorption rate of Bletilla striata glue at room temperature is 285±10%.

[0027] Compared with the prior art, the present invention has the following technical effects:

[0028] The spray film prepared by the invention has remarkable wound repairing effect, and the preparation method is simple and the raw materials used are easy to obtain.

[0029] A spray film preparation with the effect of promoting wound repair is made from cockroach extract and white schizonepeta gum. The spray film preparation can accelerate wound repair, and at the same time has the effects of stopping bleeding, reducing swelling and relieving pain, inhibiting excessive wound repair and reducing scar hyperplasia. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] For ease of explanation, the present invention is described in detail with reference to the following specific embodiments and the accompanying drawings.

[0031] Figure 1 This is the positive ion mode diagram of the cockroach extract in Example 1.

[0032] Figure 2 This is the negative ion pattern diagram of the cockroach extract in Example 1.

[0033] Figure 3 This is a three-dimensional effect diagram of the star point-response surface design of Example 2.

[0034] Figure 4 This is the Fourier infrared spectrum of Example 3.

[0035] Figure 5 This is a film formation micrograph of Example 3.

[0036] Figure 6 This is the wound healing diagram of Example 4.

[0037] Figure 7 This is the HE staining picture of the wound surface in Example 4. DETAILED DESCRIPTION

[0038] The following are specific embodiments of the present invention and the accompanying drawings to further describe the technical solutions of the present invention, but the present invention is not limited to these embodiments; in the following description, specific details such as specific configurations are provided only to help fully understand the embodiments of the present invention. Therefore, it should be clear to those skilled in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present invention.

[0039] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features of the embodiments may be combined with each other.

[0040] Unless otherwise specified, the materials, practices and experimental equipment involved in the implementation cases of the present invention are all in line with commercially available products in the relevant chemical and biotechnology fields.

[0041] Example 1: Material composition of the main drug of the film spray

[0042] A drug combination for wound repair, comprising the following components in weight percentage: 4.0%-6% of PVPK30, 1.0%-3.0% of P188, 0.05%-0.2% of benzoic acid preservatives, 3.0%-5.0% of glycerol, 0.5%-1.0% of white schizonepeta gum, and 3.0%-6.0% of cockroach extract.

[0043] The preparation method of the cockroach extract is as follows: the crushed cockroach medicinal material is placed in an extraction tank, 95% ethanol is added at a material-liquid ratio of 1:4-5, the extraction is carried out under stirring and reflux at 65°C for 2.5 hours, filtered through a 100-200 mesh screen, the filtrate is collected, and the extraction is repeated 2-3 times. The obtained filtrate is concentrated under reduced pressure at 65°C, the negative pressure value is 0.02-0.05MPa, and the concentrated solution is concentrated to 15% of the original volume, and defatted oil with a volume twice that of the concentrated solution is added to extract and defatted the concentrated solution, and the defatted liquid is filtered through a filter cloth with a mesh size of 3000-4000, and the filtrate is concentrated under reduced pressure to an extract to obtain the cockroach extract, and the yield of the extract is about 5%-7% of the medicinal material.

[0044] The cockroach extract is obtained from cockroaches by biological extraction process. The main chemical components are shown in LC-TOF / MS Figure 1-2 , and the following Table 1:

[0045]

[0046]

[0047] The preparation method of the white schizonepeta glue is as follows:

[0048] Place fresh tubers of Bletilla striata in a grinder and add deionized water at a solid-liquid ratio of 1:4-5, grind into slurry, heat the slurry at 55-70℃ for 1.5 hours, centrifuge and filter, collect the filtrate, and extract repeatedly 2-3 times. The filtrate is concentrated to 30% of the original volume at 60-70℃ under reduced pressure, ethanol is added and the ethanol concentration is adjusted to 80%-85%, and allowed to stand overnight at 0-5℃, centrifuged for 15 minutes, the supernatant is removed, the precipitate is collected, and freeze-dried to obtain Bletilla striata glue.

[0049] Further preferably, the centrifugal speed is 3000-4000 rpm.

[0050] The invention adopts modern extraction and purification technology to obtain white radix glue, and adopts HPLC-RID method to detect the molecular weight of white radix glue, and adopts drying constant weight method to detect the water absorption rate of white radix glue. The results show that the molecular weight of white radix glue is 3.5×10 4 -8.7×10 4 Da; the moisture absorption rate of Bletilla striata glue at room temperature is 285±10%.

[0051] Example 2: Optimization, screening and preparation method of composite spray film formulation

[0052] A method for preparing a drug combination for wound repair, comprising the following steps:

[0053] S1. Add an appropriate amount of 45-65°C sterilized water to the Bletilla striata gum to completely dissolve it to obtain solution 1, and cool it;

[0054] S2, dissolve PVPK30 in an appropriate amount of sterile water and stir evenly to obtain solution 2;

[0055] S3, adding P188 to the solution 2 obtained in step S2, stirring evenly to obtain solution 3;

[0056] S4, adding a benzoic acid preservative to the solution 3 obtained in step S3, and stirring to obtain a solution 4;

[0057] S5, after the temperature of the solution 1 obtained in step S1 is cooled to room temperature, the solution 4 obtained in step S4 is added to the solution 1 and stirred evenly to obtain a solution 5;

[0058] S6, adding the cockroach extract and glycerol to the solution 5 obtained in step S5, adding sterile water to make up to volume and stirring evenly, and filling to obtain the cockroach alveoli drug combination.

[0059] 1. Screening of spray film excipients

[0060] 1.1 Screening of film-forming materials

[0061] Take an appropriate amount of film-forming materials (PVPK30, PVA, BSP, P407, P188, HPMC, carbomer), add an appropriate amount of sterile water to dissolve, and dilute to 20mL. After complete dissolution, observe the dissolution and solution state, take the spraying of the liquid as the evaluation index, spray at a distance of 15cm from the white cardboard, observe the spraying effect, and record the spraying area; use a glass rod to drop the spraying solution on the slide, spread it evenly, place it at 37℃ to dry the film, and observe the film formation. In the experiment, it should be ensured that each sample is sprayed with the same spray bottle to reduce the experimental error. The preliminary investigation results of the dosage of different film-forming materials and the individual film-forming conditions are shown in Table 2. From the screening results, it can be seen that the viscosity of the film-forming material carbomer is too large after dissolution, and it is not suitable for spraying; PVA has a very low solubility and a very thin film. BSP, P188, P407, PVPK30, and HPMC can all form sprays and films, but P407 dissolves slowly and has more bubbles. The film-forming effects of the above film-forming materials alone are not ideal, and the mixed film-forming situation can be further investigated. The mixed dosage of the film-forming materials BSP, PVP-K30, P188, and HPMC can be further investigated.

[0062] Table 2 Film formation of different film-forming materials

[0063]

[0064]

[0065] 1.2 Screening of mixed film-forming materials

[0066] According to Table 2, take appropriate amount of the above-mentioned film-forming materials (PVPK30, BSP, P188, HPMC), add appropriate amount of sterilized water to dissolve, and adjust the volume to 100mL. After complete dissolution, observe the dissolution and solution state, take the spraying of the liquid as the evaluation index, spray at a distance of 15cm from the white cardboard, observe the effect of the spray, and record the spraying area; use a glass rod to drop the spray solution on the slide, spread it evenly, place it at 37℃ to dry and form a film, and observe the film formation. In the experiment, it should be ensured that each sample is sprayed with the same spray bottle to reduce the experimental error. The experimental results are shown in Table 3. From the experimental results, it can be seen that "5.0% PVPK30 + 3.0% P188 + 0.5% BSP" has better film-forming properties than other mixed film-forming materials, and has better effects in solution state, spray state, and film-forming evaluation. Therefore, it is feasible to combine PVPK30, P188 and BSP into a mixed film-forming material, which is the preferred mixed film-forming material in this study, and its compatibility ratio can be further optimized to obtain the best film-forming material.

[0067] Table 3 Screening of mixed film-forming materials

[0068]

[0069]

[0070] 1.3 Selection of plasticizer

[0071] Plasticizers can increase the toughness of the film. Commonly used plasticizers are glycerol, propylene glycol, etc. Glycerol not only has good safety and water solubility, but also has a good moisturizing effect. The film-forming materials selected in this study are all water-soluble film-forming materials, which create favorable conditions for wound healing, so glycerol is selected as a plasticizer.

[0072] 1.4 Choice of preservatives

[0073] Different amounts of sodium ethylparaben, sodium hydroxybenzoate and sodium benzoate were added to the spray, and colloidal substances were observed to precipitate, as shown in Table 4. The reason may be that metal ions chelate with BSP to precipitate colloids, so metal ion-containing preservatives such as sodium ethylparaben, sodium hydroxybenzoate and sodium benzoate cannot be used as preservatives for the spray. When different amounts of benzoic acid were added, the stability of the solution was not destroyed, and a uniform solution could still be obtained. Therefore, benzoic acid is the best preservative for the spray, and the acidic condition can effectively inhibit the growth of microorganisms.

[0074] Table 4 Preservative screening

[0075]

[0076] 1.5 Preparation of blank spray solution

[0077] Weigh an appropriate amount of BSP, add an appropriate amount of sterile water to dissolve it, then add PVPK30, P188, plasticizer, and preservative in sequence, stir to dissolve and mix evenly, add sterile water to make up to volume and mix well.

[0078] 2. Single factor experimental design of film-forming excipients

[0079] 2.1 Evaluation indicators

[0080] The evaluation indicators are spray effect, film-forming time and film performance. The specific evaluation standards are as follows: spray at a distance of 15 cm from the white cardboard, observe the spray effect, and calculate the spray area; take an appropriate amount of spray solution, evenly apply it on the slide, put it in a 37°C oven, and consider it to be a film after the surface is completely dry, and record the film-forming time; evaluate the film performance by observing the transparency, uniformity, thickness, presence of bubbles and toughness of the film.

[0081] 2.2 Investigation of PVPK30 dosage

[0082] The types and amounts of other excipients in the fixed prescription were used to prepare the spray film with 4.0%, 4.5%, 5.0%, 5.5%, and 6.0% PVPK30, respectively. The volume was fixed to 20mL and mixed. The results are shown in Table 5. When the other excipients remained unchanged, with the increase of PVPK30 dosage, the solution gradually precipitated, the thickness of the film increased, the transparency of the film decreased, and the spray area became smaller. When the dosage increased to 5.5%, the spray effect was in a semi-spray state. When the dosage was 6.0%, it was sprayed in a dispersed linear state, and it was difficult to form a spray; the film-forming time showed a trend of first decreasing and then increasing with the increase of dosage. Too much dosage will inevitably increase the film-forming time, but when the dosage is too small, it may be that the content of the film-forming material is too small, resulting in a too thin solution, which is not conducive to drying to form a film. The dosage of PVPK30 has no significant effect on the viscosity of the film. Except for the poor toughness of the film formed when the dosage is too small and too thin, the film viscosity of the other dosages is good. Therefore, considering the spray area, film-forming time, solution state and film-forming performance, the optimal dosage of PVPK30 is about 5.0%.

[0083] Table 5 Effect of PVPK30 dosage on spray film formulation

[0084]

[0085] 2.3 Investigation of BSP usage

[0086] According to the previous film-forming condition and dosage experimental results of Bletilla striata, BSP was added as much as possible without affecting the quality of the spray film. Therefore, 0.5%, 1.0%, 1.5%, 2.0%, and 2.5% of BSP were added respectively. In addition to the indicators, the state of the solution should be observed. The change of water content was detected by Fourier infrared spectroscopy. The determination method and moisturizing effect evaluation method were the same as those of glycerol. The results are shown in Table 6; the comprehensive influence results are shown in Table 7. BSP has a significant effect on the state of the solution, spray effect, spray area, film-forming time, film-forming performance and moisturizing effect. With the increase of BSP dosage, the solution gradually precipitates colloidal substances, the spray effect becomes worse, the spray area decreases, the film-forming time is also prolonged, the film-forming performance is also worse, and the moisturizing effect also changes accordingly. However, when the BSP dosage is about 0.8%, the film has a better effect, which is conducive to the formation of the film and moisturizing of the wound surface.

[0087] Table 6 Effect of BSP on moisturizing effect of spray film formulation

[0088]

[0089] Table 7 Effect of BSP dosage on spray film formulation

[0090]

[0091]

[0092] 2.4 Investigation of P188 dosage

[0093] The types and amounts of other excipients in the prescription were fixed. 1.0%, 1.5%, 2.0%, 2.5%, and 3.0% of P188 were used to prepare the spray film, and the volume was fixed to 20 mL. The results are shown in Table 8. As the amount of P188 increased, the spray area decreased, indicating that the amount of P188 also affected the spray effect. When the amount was 2.5%, the solution would produce a small amount of bubbles and spray in a semi-spray state, with a spray area of ​​54.35 cm 2 ; The amount of P188 also has a significant effect on the film-forming time. When the amount of P188 is 2.0%, the film-forming time is relatively short. Secondly, the amount of P188 also affects the solution state, spray area, and film-forming performance. As the amount changes, the solution state, spray area, and film-forming performance all change accordingly. When the amount is too small or too large, the film-forming property is poor. Therefore, considering the spray area, film-forming time, and film-forming performance, the amount of P188 is about 2.0%, which is the best amount.

[0094] Table 8 Effect of P188 dosage on spray film formulation

[0095]

[0096] 2.5 Investigation of glycerol dosage

[0097] The types and amounts of other excipients in the formula were fixed, and 1.0%, 2.0%, 3.0%, 4.0%, and 5.0% glycerol were used to prepare the spray film, and the volume was fixed to 20 ml. 5 μL of the spray film solution was taken to cover the small hole on the Fourier transform infrared spectrometer ATR, and the water content change at 0h, 0.5h, 1.0h, and 1.5h was monitored. The absorbance value (A) was used as the ordinate and the wave number (cm -1 ) was used as the horizontal axis to calculate the peak area of ​​the infrared absorption peak of hydroxyl (-OH) at each time point and the difference with the initial peak area. The results are shown in Table 9. Then the moisturizing effect was evaluated with the mean of the difference at the three time points. The larger the mean of the difference at each time point, the worse the moisturizing effect, and vice versa. The comprehensive influence results are shown in Table 10. Glycerol has little effect on the solution state, spray effect, spray area and film-forming time, but the amount of glycerol has a significant effect on the film-forming properties and moisturizing effect. When the amount of glycerol is about 4.0%, it has a better effect, which is beneficial to the formation of drug film and moisturizing of the wound surface.

[0098] Table 9 Effect of glycerin on moisturizing effect of spray film prescription

[0099]

[0100] Table 10 Effect of glycerol dosage on spray film formulation

[0101]

[0102] 2.6 Investigation of the dosage of benzoic acid

[0103] The types and amounts of other excipients in the fixed prescription were added with 0.10%, 0.15%, 0.20%, 0.25%, and 0.30% benzoic acid to prepare the spray film, and the volume was fixed to 20 ml. The pH was measured, and the results are shown in Table 11. The optimum pH of human skin is 4.5-6.5, and the antibacterial effect is best when pH < 5.0. The results show that the preservative has no obvious effect on the solution state, spray effect spray area, film formation time, and film performance, but has a greater effect on the pH of the solution. The pH of the solution decreases with the increase of the amount of benzoic acid. When 0.10% benzoic acid is added, the pH of the solution is 4.52. Therefore, the effect is best when the amount of benzoic acid is 0.10%.

[0104] Table 11 Effect of benzoic acid dosage on spray film formulation

[0105]

[0106]

[0107] 2.7 Investigation of the dosage of cockroach extract

[0108] Under the condition that the quality of the spray film is not affected, as much cockroach extract as possible is added. Therefore, 2.0%, 3.0%, 4.0%, 5.0%, and 6.0% cockroach extract are added respectively. In addition to the inspection index, the state of the solution needs to be observed. The results are shown in Table 12. The cockroach extract has no significant effect on the spray effect, film-forming time, and spray area of ​​the spray film. When the amount of cockroach extract is 5.0%, precipitation can be observed in the spray film solution. Therefore, in summary, the amount of cockroach extract is 3.0% optimal.

[0109] Table 12 Effect of the dosage of cockroach extract on the spray formulation

[0110]

[0111] 3. Optimization of the formulation composition of the film spray by central complex design-response surface optimization method

[0112] 3.1 Evaluation indicators

[0113] The comprehensive score of the film-forming spray is used as the evaluation index, and the specific scoring criteria are as follows:

[0114] (1) Spray effect: The full score for spray is 25 points. Spray at a distance of 15 cm from a white cardboard, observe the spray effect, and record the spray area. Mist spray is the best, with a score of 21-25 points, followed by semi-mist spray, with a score of 15-20, and dispersed linear spray, with a score of 10-15. Spraying with a few sparse lines is the worst, with a score of 1-10.

[0115] (2) Film-forming time: The full score for gelation time is 25 points. Take an appropriate amount of film-forming spray solution and evenly apply it on a glass slide. Place it in a 37°C oven and wait until the surface is completely dry. The gelation is considered complete and the gelation time is recorded. A gelation time of 5 min is considered a full score of 25 points. For every additional minute of gelation time over 5 min, 1 point will be reduced from 25 points.

[0116] (3) Moisturizing effect: The water content of the spray solution at 0, 0.5, 1, and 1.5 h was measured using a Fourier transform infrared spectrometer. The difference between the water peak area at each time point and the initial peak area and the average of the differences at the three time points were calculated. When the average value was 90, the full score was 25 points. Every increase in the average value by 1 point would reduce the score by 2 points from the 25-point scale.

[0117] (4) Film performance: The full score is 25 points, mainly including film maintenance time 5 points, uniformity 10 points and thickness 10 points. The film maintenance time is judged by the degree to which the film can maintain its integrity, with 8 hours as the full score and a score between 0-5 points. The film is scored based on the presence of bubbles and the uniformity of the film surface. The film is bubble-free, transparent and uniform, with a score between 5-10 points. The film has bubbles and a score between 0-5 points. After the film is formed, the film is rolled into strips. The film that can be rolled into thin threads is the best thickness and is scored between 7-10 points. The film that can be completely torn off a layer without deformation is considered too thick and is scored below 7 points. The film that cannot be rolled into thin threads is considered too thin and is scored below 7 points.

[0118] The total score of spray effect, film-forming time, film performance and moisturizing effect is added together to get the comprehensive score.

[0119] 3.2 Star point design and results

[0120] From the results of the single factor test, it can be seen that PVPK30, P188 and BSP have the most significant impact on the spray film. Therefore, PVPK30, P188 and BSP are selected as the main influencing factors of the spray film. The comprehensive score of the spray film is used as the evaluation index to optimize the prescription of the spray film. The factors and levels are shown in Table 13.

[0121] Table 13 Star point design factor level table

[0122]

[0123] According to the results of the single factor test, the comprehensive score (Y) was used as the response value, the factor level table was designed according to Central Composite, and the experimental plan was designed using the software Design-Expert 8.0.6.1. The experimental plan and results are shown in Table 14. Table 15 shows the moisturizing effect of each group.

[0124] Table 14 Star point experiment design table and results

[0125]

[0126]

[0127] Table 15 Moisturizing effect of each group in star point experimental design

[0128]

[0129]

[0130] 3.3 Model Fitting

[0131] The data results in Table 14 were processed by using Design-Expert 8.0.6.1. The results of variance analysis are shown in Table 16. Taking the comprehensive score (Y) as the response value, the regression fitting equations for PVPK30 (A), P188 (B) and BSP (C) were obtained:

[0132] Y=86+0.5227*A-0.1346*B-0.6505*C+1.06*AB-0.3375*AC+0.9875*BC-3.05*A 2 -1.03*B 2 -1.42*C 2

[0133] Further testing of the regression equation yields a table showing that the regression equation model is extremely significant (P<0.01 indicates extremely significant), and the lack-of-fit term is not significant (P>0.05 indicates not significant, and P<0.05 indicates significant), indicating that the model is reliable. Regression coefficient R 2 =0.9886(R 2 ≥0.90 is considered that the model has good correlation), as shown in Table 17, indicating that the model is reliable. From the F values ​​of factors A, B, and C, it can be seen that the F values ​​of A and C are not much different, and they are quite different from the F value of B. It can be concluded that among the three factors, PVPK30 and BSP have a greater impact on the comprehensive score of the spray film than P188. According to the P value of the variance analysis of each experimental factor, it can be seen that the significance of the impact of each factor on the comprehensive score of the spray film is: factor A>factor C>factor B.

[0134] Table 16 Response surface model variance analysis results

[0135]

[0136]

[0137] Table 17 ANOVA results

[0138]

[0139] 3.4 Star-site design-response surface methodology optimization and predictive evaluation

[0140] According to the fitted quadratic polynomial model, the software Design-Expert 8.0.6.1 was used to draw a three-dimensional effect diagram and a two-dimensional contour diagram to screen out the optimal prescription composition. The influence of each factor on the response value is shown in Figure 3 .like Figure 3 As shown in the three-dimensional effect diagram, the steeper the slope of the response surface and the larger the color span, the more significant the interaction between the factors; in the two-dimensional contour map, the more elliptical the shape, the more significant the interaction. The results showed that PVPK30, P188 and BSP all had an effect on the spray agent, and with the increase in the dosage of the three factors, the comprehensive score of the spray agent had a trend of first increasing and then decreasing. It was concluded that PVPK30 and BSP had a greater impact on the comprehensive score of the spray agent, and P188 had the least impact. Through the analysis of the software Design-Expert 8.0.6.1, the optimal prescription of the spray agent was obtained: PVPK30 was 5.03644g, P188 was 1.91566g, and BSP was 0.675987g ​​per 100ml. Under this prescription condition, the predicted value of the comprehensive score was 86.1229.

[0141] In order to facilitate weighing, the optimal prescription process was revised to: PVPK30 accounted for 5.0%, P188 accounted for 2.0%, benzoic acid preservatives accounted for 0.1%, glycerin accounted for 4.0%, white schizonepeta gum accounted for 0.70%, and cockroach extract accounted for 3.0%.

[0142] 3.5 Verification of the optimal prescription

[0143] The optimal extraction process was used for confirmation experiments. The results are shown in Table 18. The actual comprehensive score value is similar to the predicted comprehensive score value. The results show that the prescription conditions are true, reliable and feasible.

[0144] Table 18 Optimal prescription verification and results

[0145]

[0146] Example 3: Physical and chemical characterization of composite film-forming agent

[0147] 1. Fourier transform infrared spectroscopy characterization

[0148] The infrared spectrum was analyzed using a Fourier transform infrared spectroscopy (FTIR) analyzer with a resolution of 4 cm -1 , the number of scans was 16, and the spectral range was 4000~500cm -1 The result is as follows Figure 4 As shown, the spray film agent has a wave number of 3305cm －1 There is a strong OH stretching vibration characteristic absorption peak at 2880cm －1 There is a characteristic absorption peak of CH at 1651cm －1 There is a strong and narrow carbonyl C=O bending vibration absorption peak at 1105cm －1 There is a characteristic absorption peak of furanose at 1200~1000cm -1 There are strong absorption peaks of COH and COC stretching vibration at 841 cm -1 There is a characteristic absorption peak of α-type glycosidic bond.

[0149] 2. Microscopic Characterization

[0150] The prepared spray film was sprayed on a glass slide, placed in a 37°C oven for drying, and the film formation was observed under an optical microscope and photographed for record. Figure 5 As shown, the spray film is uniform and smooth without bubbles, the drug is evenly dispersed, and the thickness is moderate, indicating that the prescription and preparation process are better.

[0151] Example 4: Wound healing effect experiment

[0152] 1. Animal Model Establishment

[0153] All rats were adaptively fed for 1 week, and their back hair was shaved. Sodium pentobarbital (0.3 mL / 100 g) was injected intraperitoneally for anesthesia. The surgical area was locally disinfected with medical iodine. Except for 12 rats in the normal group, circular marks with a diameter of 1.0 cm were drawn on the hair removal sites on both sides of the dorsal spine of the remaining rats. Ophthalmic scissors were used to trim the skin along the predetermined line to the surface of the muscle fascia (full-thickness skin defect wound), resulting in two wounds of similar size. Sterile cotton balls were then used to stop bleeding and iodine was used for disinfection. Natural exposure was maintained throughout the operation.

[0154] 2. Grouping and Dosing

[0155] The rats with full-thickness skin defect wounds after successful modeling were divided into model group, blank spray film group, Kangfuxin liquid group, and spray film group, with 12 rats in each group. The normal control group and model group were sprayed with normal saline to cover the wound every day, the blank matrix group was sprayed with an equal amount of blank spray film every day, the Kangfuxin liquid group was sprayed with an equal amount of Kangfuxin liquid, and the spray film group was given an equal amount of spray film obtained in this study. Photos were taken on the 0th, 3rd, 7th, 10th, and 14th days after modeling, and the wound healing rate was calculated.

[0156] 3. Observation of Physiological Activities of Rats

[0157] The general conditions of the rats in each group, such as mental state, body shape, food intake, and weight changes, were observed and recorded daily. The normal group had smooth white fur, good mental state, lively and active, and normal diet and bowel movements; the model group had less food and drink, slightly loose stools, less shiny yellow fur, a listless mental state, buried their heads in the litter, and slow weight growth; the fur color and glossiness of the Kangfuxin liquid group and the spray film group were improved compared with the model group.

[0158] 4. Sampling and processing

[0159] Six rats were randomly selected from each group at 7 and 14 days after administration, anesthetized with isoflurane, and the wound tissue with a diameter of 1.5 wounds and a size of circular wound with the wound center as the center was cut aseptically. Part of the tissue was immersed in 4% paraformaldehyde solution for pathological sectioning, and the other part of the tissue was quickly frozen in liquid nitrogen and transferred to a -80℃ refrigerator for storage.

[0160] 5. Observation of wound healing

[0161] The wound images of rats in each group were taken on the 0th, 3rd, 7th, 10th and 14th days after modeling. The wound healing results are shown in Figure 6 At 0 days after modeling, no obvious abnormalities were found in the wounds of the rats in the model group, blank matrix group, Kangfuxin liquid group, and spray film group. At 3 days after modeling, purulent secretions appeared at the edge of the wounds of the rats in the model group, and edema was severe. No moistening phenomenon was found in the blank matrix group, the wounds of the Kangfuxin liquid group were slightly scabby and the wounds shrank, and the wounds of the spray film group were significantly shrunk and obvious scabs formed without edema. At 7 days after modeling, the hair around the wounds of the model group was dull and yellow, and the wounds were inflamed and edematous. Slight scabby phenomenon was found in the blank matrix group, the wounds of the Kangfuxin liquid group were shrunk but with visible signs of blood color, and the wounds of the spray film group were Well-grown granulation tissue was seen, the scab disappeared, and the wound healing was almost complete as seen with the naked eye; on the 10th day after modeling, the wounds in the model group, blank matrix group, and Kangfuxin liquid group were further reduced, while the wounds in the spray film group had healed significantly and completely, with obvious new granulation tissue visible; on the 14th day after modeling, the wound area in the model group was still relatively large, with purulent secretions still discharged from the edge of the wound, the wounds in the Kangfuxin liquid group were completely healed but obvious scar hyperplasia was visible, while the wounds in the spray film group were completely repaired, with no obvious difference between the new skin around the wound and normal skin tissue, and the new fur covered the healed wound without obvious scar remaining.

[0162] 6. HE staining and fibroblast counting

[0163] The wound skin tissue placed in 4% paraformaldehyde solution was dehydrated, embedded, sliced, and stained with hematoxylin-eosin to observe the changes in cell nuclei, cytoplasm, collagen fibers, inflammatory cell infiltration, and local blood vessels. Figure 7 On the 7th day of modeling, no clear skin structure was seen in the model group, a large number of cells in the epidermis were necrotic, the nuclei were fragmented or disappeared, the skin was condensed and darkly stained, the spinous layer structure disappeared, the collagen fibers in the dermis were sparse, no skin appendages were seen, and a large number of inflammatory cells infiltrated the tissue; in the blank matrix group, large areas of necrosis, nuclear fragmentation, sparse collagen fibers, obvious inflammatory cell infiltration were seen, and no skin appendages were seen; in the Kangfuxin liquid group, no skin appendages were seen in the visual field, obvious inflammatory cell infiltration and, collagen fibers and fibroblasts were not obvious; in the spray film group, obvious scabs were seen in the epidermis in the visual field, a large number of cells were necrotic, the nuclei were fragmented, the skin was condensed and darkly stained, the collagen fibers in the dermis were abundant, a large number of inflammatory cells infiltrated and blood vessels were seen in the tissue, and the epidermis was initially formed. On the 14th day of modeling, the epidermis disappeared in the visual field of the model group and the blank matrix group, the dermis was exposed, the collagen fibers were lost, no skin appendages were seen, a large number of inflammatory cells infiltrated, a small number of cells were necrotic, the nucleus was fragmented, and the skin appendages were darkly stained; in the Kangfuxin liquid group, the cytoplasm was red-stained, the epidermis was partially shed, the collagen fibers in the dermis were partially lost, no skin appendages were seen, and the stratum spinosum was not obvious; in the spray film group, complete skin appendages were seen, the stratum corneum was obvious, the epidermis structure was intact, the stratum spinosum was slightly thickened, the local spinous processes were elongated, the dermis was rich in collagen fibers, and the skin appendages hair follicles and sebaceous glands were visible.

[0164] In summary, the preparation process of the spray film is feasible, the wound repair effect is significant, and it has the effect of inhibiting excessive wound repair and scar hyperplasia.

[0165] Those skilled in the art to which the present application belongs may make various modifications or additions to the specific embodiments described or replace them in a similar manner, but they will not deviate from the inventive concept of the present application or exceed the scope defined by the attached claims.

Claims

1. A drug combination for wound repair, characterized in that: The invention comprises the following components in weight percentage: PVPK30 accounts for 4.0%-6%, P188 accounts for 1.0%-3.0%, benzoic acid preservatives account for 0.05%-0.2%, glycerin accounts for 3.0%-5.0%, white jelly gum accounts for 0.5%-1.0%, and cockroach extract accounts for 3.0%-6.0%.

2. A drug combination for wound repair according to claim 1, characterized in that: The cockroach extract comprises 11 kinds of amino acids, 9 kinds of alkaloid components, 6 kinds of fatty acid components, 5 kinds of nucleoside components, 4 kinds of peptide components, 4 kinds of phospholipid components and 4 kinds of sugar components.

3. A drug combination for wound repair according to claim 2, characterized in that: The amino acids are C7H 13 NO3、C9H 11 NO3, C 12 H 23 NO7, C5H9NO2, C5H 10 N2O3, C6H9N3O2, C6H 14 N4O2、C9H 11 NO4, C 11 H 12 N2O3、C 10 H7NO3, C5H9NO4, the alkaloid components are C8H9NO, C 25 H 47 NO4, C5H4N4O3, C 11 H 12 N2, C8H10N4O2, C5H 15 NO4P、C9H 17 NO4, C6H6N2O, C7H 15 NO3, the fatty acid components are 11 H 12 O4、C 22 H 43 NO, C 18 H 34 O2, C8H8O2, C 13 H 24 O4, C7H6O4, the nucleoside components are C4H4N2O2, C 11 H 15 N5O3S、C 11 H 15 N5O5、C 12 H 13 ClN4、C9H 13 N2O9P, the peptide components are C 10 H 20 N2O4、C9H 18 N2O3、C 14 H 24 N4O6、C7H 16 N4O2, the phospholipid components are C 44 H 80 NO8P、C 26 H 52 NO7P, C3H9O6P, C8H 20 NO6P, the sugars are C 17 H 22 O 10 、C2H8NO4P、C 12 H 24 O 11 、C 18 H 32 O 16 。 4. A drug combination for wound repair according to claim 3, characterized in that: The cockroach extract also includes C6H6O2, C 15 H 10 O4、C8H 14 O2S2、C7H6O5、C 37 H 70 O5, C3H9N.

5. A drug combination for wound repair according to claim 1, characterized in that: The preparation method of the white schizonepeta glue is as follows: Place fresh tubers of Bletilla striata in a grinder, add deionized water at a solid-liquid ratio of 1:4-1:5, grind into slurry, heat the slurry at 55-70℃ for 1.5-2.0 hours, centrifuge and filter, collect the filtrate, extract repeatedly 2-3 times, place the filtrate at 60-70℃ under reduced pressure and concentrate to 30% of the original volume, add ethanol and adjust the ethanol concentration to 80%-85%, stand at 0-5℃ overnight, centrifuge for 15 minutes, remove the supernatant, collect the precipitate, freeze-dry to obtain Bletilla striata glue.

6. A drug combination for wound repair according to claim 1, characterized in that: The preparation method of the cockroach extract is as follows: The crushed cockroach medicinal material is placed in an extraction tank, 80%-95% ethanol is added at a solid-liquid ratio of 1:4-5, and the mixture is stirred and refluxed at 65°C for 2.5 hours, filtered through a 100-200 mesh sieve, the filtrate is collected, and the extraction is repeated 2-3 times. The obtained filtrate is concentrated under reduced pressure at 65°C with a negative pressure value of 0.02-0.05MPa to 10%-15% of the original volume, and defatted oil with a volume 2-3 times that of the concentrated solution is added to extract and defatt the concentrated solution. The defatted liquid is filtered through a 3000-4000 mesh filter cloth, and the filtrate is concentrated under reduced pressure to an extract state with a specific gravity of 1.20-1.35, thereby obtaining a cockroach extract, and the yield of the extract is about 5%-7% of the medicinal material.

7. A method for preparing the pharmaceutical combination according to any one of claims 1 to 6, characterized in that: The specific steps include: S1. Add an appropriate amount of 45-65°C sterilized water to the Bletilla striata gum to completely dissolve it to obtain solution 1, and cool it; S2, dissolve PVPK30 in an appropriate amount of sterile water and stir evenly to obtain solution 2; S3, adding P188 to the solution 2 obtained in step S2, stirring evenly to obtain solution 3; S4, adding a benzoic acid preservative to the solution 3 obtained in step S3, and stirring to obtain a solution 4; S5, after the temperature of the solution 1 obtained in step S1 is cooled to room temperature, the solution 4 obtained in step S4 is added to the solution 1 and stirred evenly to obtain a solution 5; S6, adding the cockroach extract and glycerol to the solution 5 obtained in step S5, adding sterile water to make up to volume and stirring evenly, and filling to obtain the cockroach alveoli drug combination.

8. The method for preparing the drug combination according to claim 7, characterized in that: The dosage form of the drug combination is a composite spray film, and the composite spray film is packed into a spray bottle.

9. The method for preparing the drug combination according to claim 7, characterized in that: The molecular weight of the white schizonepeta gel in step S1 is 3.5×10 4 -8.7×10 4 Da, the moisture absorption rate of Bletilla striata glue at room temperature is 285±10%.

10. Use of the drug combination according to any one of claims 1 to 6 or the drug combination prepared by the preparation method according to claim 9 in preparing a drug for wound repair, characterized in that: The drug also includes other pharmaceutically acceptable excipients.

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