Oriental beauty tea extract and application thereof in medical dressing

Abstract

The invention discloses an oriental beauty tea extract and application thereof in medical dressings, and belongs to the technical field of dressing production.The oriental beauty tea extract is obtained by extracting oriental beauty tea powder through cellulase and hydroxypropyl-beta-cyclodextrin and then conducting freeze drying, and then the oriental beauty tea extract is obtained through freeze drying. The dressing is prepared by mixing modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and a beta-cyclodextrin inclusion compound. The technical barrier of industrialization of tea polyphenol dressings is fundamentally overcome through a three-stage synergistic mechanism of molecular pre-packaging, physical barrier and chemical bonding, and a research direction is provided for development of long-acting, stable and intelligent-response high-end functional dressings.

Description

Technical Field

[0001] The invention belongs to the technical field of dressing production, and particularly relates to an Oriental Beauty Tea extract and application thereof in medical dressings. Background Art

[0002] Oriental Beauty Tea is a specialty tea in my country, rich in polyphenols, flavonoids, and natural antioxidants, and holds great promise in the field of dressings. Traditional dressings (such as gauze and hydrogels) often incorporate plant extracts to enhance their antibacterial properties, but adding tea extracts to traditional dressings presents significant drawbacks: (1) Active ingredients are easily inactivated: high temperature sterilization or long-term storage leads to polyphenol oxidation; (2) Poor sustained-release effect: The tea extract release rate is too fast (>80% released within 2 h), and it cannot maintain a long-term effect; (3) Insufficient compatibility: Tea extract is easy to precipitate after mixing with dressing base material (such as chitosan), affecting uniformity and efficacy.

[0003] The above defects seriously restrict the application of Oriental Beauty Tea in high-end medical dressings. Summary of the Invention

[0004] The purpose of the present invention is to provide an Oriental Beauty Tea extract and its application in medical dressings, so as to solve the defects of existing dressings when adding tea extracts.

[0005] The purpose of the present invention can be achieved through the following technical solutions: An Oriental Beauty Tea extract is prepared by the following steps: Oriental Beauty tea is dried and pulverized through a 65-mesh sieve to obtain tea powder, an extract is added to the tea powder, and then cellulase and hydroxypropyl-β-cyclodextrin are added thereto. The system is heated to 45-50°C and subjected to constant temperature extraction for 20-60 minutes. After extraction, the system is centrifuged at 4500 rpm for 10 minutes, the supernatant is rotary evaporated, and then freeze-dried to a powder to obtain the Oriental Beauty tea extract.

[0006] Furthermore, the extract is an ethanol solution with a volume fraction of 20% to 80%.

[0007] Furthermore, the usage ratio of the tea powder, the extract, the cellulase and the hydroxypropyl-β-cyclodextrin is 2g:10mL:0.05-0.25g:0.1-1.0g.

[0008] During the preparation of the extract, the present invention adds cellulase and hydroxypropyl-β-cyclodextrin. Cellulase can destroy tea cell walls, increasing the dissolution rate of active ingredients such as polyphenols. Hydroxypropyl-β-cyclodextrin is a green, synthetic, alkylated derivative of β-cyclodextrin, characterized by a hydrophilic outer edge, a hydrophobic inner cavity, and high solubility. This improves the dissolution rate and stability of polyphenols, providing initial protection during the extraction stage and reducing oxidative inactivation. Furthermore, the present invention optimizes the conditions of an ethanol solution volume fraction of 20% to 80%, a temperature of 45 to 50°C, and an extraction time of 20 to 60 minutes to achieve a synergistic effect between cellulase and hydroxypropyl-β-cyclodextrin.

[0009] Furthermore, the application of the Oriental Beauty Tea extract in medical dressings comprises the following steps: S1. Dissolve β-cyclodextrin in an ethanol solution at 58-60° C. to obtain a β-cyclodextrin mixed solution, dissolve the Oriental Beauty Tea extract in anhydrous ethanol to obtain an extract mixed solution, and after the β-cyclodextrin mixed solution is cooled to 45-50° C., add the extract mixed solution to the β-cyclodextrin mixed solution to obtain a mixed solution, place the mixed solution in the dark, and stir at a constant temperature for 10-30 minutes. After completion, place the mixed solution in a low-temperature environment at 4° C. and let it stand overnight to precipitate crystals, collect the solid component by filtration, and then wash the solid component with anhydrous ethanol and vacuum dry to obtain a β-cyclodextrin inclusion compound; S2. Add modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion complex into pure water with stirring to obtain a mixed solution, then add 1.0 M sodium hydroxide solution thereto with stirring, stir at room temperature for 3 to 5 minutes, let stand for 30 minutes, and sterilize after completion to obtain a gel dressing.

[0010] During the preparation of the above-mentioned dressing, the present invention forms an inclusion complex with the Oriental Beauty Tea extract and β-cyclodextrin. The inclusion process is carried out in a dark, low-temperature (4°C) environment to avoid photothermal degradation. The inclusion complex "encapsulates" the active ingredient in a hydrophobic cavity, isolating oxygen and moisture, and significantly enhancing the stability of the polyphenol active ingredients in the Oriental Beauty Tea extract; then, the present invention uses sodium alginate and hyaluronic acid modified with 3-aminophenylboronic acid (step S2). The boronic acid group forms a dynamic covalent bond with the ortho-diphenol hydroxyl group of the tea polyphenols, improving compatibility and reducing precipitation. At the same time, the boronic acid bond is pH-responsive, achieving sustained release, and polyvinyl alcohol is used as an auxiliary gelling agent to enhance the mechanical strength of the dressing. Overall, the present invention forms a coherent protection chain (enzyme-assisted extraction → cyclodextrin inclusion → borated substrate bonding) through the three-step process of extraction, inclusion, and dressing preparation, systematically solving the problems of inactivation, rapid release, and precipitation.

[0011] Furthermore, the ethanol solution in S1 is prepared by mixing pure water and anhydrous ethanol in a volume ratio of 2:1; the dosage ratio of the β-cyclodextrin, ethanol solution, Oriental Beauty Tea extract, and anhydrous ethanol is 5g:100mL:2g:40-50mL.

[0012] Furthermore, the usage ratio of pure water, modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0M sodium hydroxide solution in S2 is 200mL:12-15g:12-15g:2-3g:4-5g:10mL.

[0013] Furthermore, the modified sodium alginate described in S2 is prepared by the following steps: Sodium alginate and 3-aminophenylboronic acid monohydrate are dissolved in pure water, and then carbodiimide hydrochloride is added thereto with stirring to obtain a mixed solution. The mixed solution is stirred at room temperature for 24 hours. After completion, it is dialyzed using a dialysis bag with a molecular weight cutoff of 8000 to 14000 Da for 3 days, and finally freeze-dried to obtain modified sodium alginate.

[0014] Furthermore, the usage ratio of the sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride is 10g:3.9-4.0g:1L:9.6-10.0g.

[0015] Furthermore, the modified hyaluronic acid described in S2 is prepared by the following steps: Hyaluronic acid was dissolved in a mixed solvent, and 3-aminophenylboronic acid monohydrate and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride were added thereto with stirring. 1M sodium hydroxide solution was then added thereto with stirring to adjust the pH of the system to 6.4-6.8. After completion, the system was stirred and reacted at room temperature for 48 hours. After completion, the system was dialyzed for 4 days using a dialysis bag with a molecular weight cutoff of 8000-14000 Da, and the solid component was collected and freeze-dried to obtain modified hyaluronic acid.

[0016] Furthermore, the mixed solvent is prepared by pure water and dimethylformamide in a volume ratio of 3:2; the usage ratio of the hyaluronic acid, mixed solvent, 3-aminophenylboronic acid monohydrate, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride is 5g:100mL:1.56~1.60g:3.46~3.50g.

[0017] Beneficial effects of the present invention: The present invention provides an Oriental Beauty Tea extract and its application in medical dressings. The present invention is directed to the extraction stage of the Oriental Beauty Tea extract, the inclusion stage in the dressing preparation process, and the dressing forming stage. Firstly, the extraction stage, the present application solves the initial stability problem of phenolic substances in Dongfangmeiren tea extract through enzyme-cyclodextrin synergistic effect. The specific principle is that cellulase is used to hydrolyze tea leaf cell wall cellulose to release embedded polyphenols (improve dissolution rate); the hydrophobic cavity of hydroxypropyl-β-cyclodextrin (HP-β-CD) wraps the polyphenol hydrophobic group to form a pre-inclusion compound, which isolates oxygen (reduces oxidation during extraction). The present application first introduces HP-β-CD pre-protection in the extraction stage to inhibit the inactivation of phenolic substances from the source.

[0018] Data verification (Table 1): Comparative Example 3 (lack of HP-β-CD): retention rate is only 64.7% → lack of pre-protection leads to partial oxidation of initial polyphenols. Comparative Example 4 (without enzyme + HP-β-CD): retention rate is 60.2% → loss of active ingredients in crude extract is greater.

[0019] Then, the inclusion stage, the present application solves the release and storage inactivation problem of Dongfangmeiren tea extract by using β-cyclodextrin double encapsulation. The specific principle is that in the β-cyclodextrin inclusion (S1), the present application encapsulates the extract in the hydrophobic cavity again under dark and low temperature (4℃), completely isolating water / oxygen. The low-temperature light-avoiding inclusion process of the present application avoids photothermal degradation and strengthens the encapsulation integrity.

[0020] Data verification (Table 1): Comparative Example 5 (directly adding extract): 2-hour release rate is 88.6%, retention rate is 52.3% → no encapsulation leads to rapid release and oxidation; Example 5 (complete inclusion): 2-hour release rate is 28.5%, retention rate is 93.2% → encapsulation significantly delays release and blocks oxidation.

[0021] Next, the dressing forming stage, the present application solves the compatibility and intelligent release problem of Dongfangmeiren tea extract and gel base material by using boric acid dynamic bonding. The specific principle is that during the boric acid modification (S2-S3) process, the present application uses sodium alginate / hyaluronic acid to graft 3-aminophenyl boronic acid to form a boronic acid group, and the boronic acid group forms a pH-responsive borate ester bond (dynamic covalent bond) with the ortho-diphenol hydroxyl group of polyphenol. Finally, it achieves dual effects: suppress precipitation: dynamic bonding reduces polyphenol free, inhibits phase separation; intelligent release: the key dissociation triggered by the slightly acidic environment (pH≈5.5) of the wound realizes on-demand release. The present application first introduces boric acid dynamic chemical bond into tea polyphenol dressing, synchronously solving the compatibility and release problems.

[0022] Data verification (Table 1): Comparative Example 6 (lack of modified sodium alginate): precipitation amount is 21.7%, 12-hour release rate is 87.9% → lack of boric acid bond leads to increased precipitation and accelerated release; Example 5 (complete bonding): precipitation amount is 2.1%, 12-hour release rate is 74.9% → dynamic bonding inhibits precipitation and cooperatively releases.

[0023] In summary, compared to existing technologies, this invention pioneers a novel protection chain architecture: extraction (enzyme + HP-β-CD pre-protection) → inclusion (β-CD secondary encapsulation) → molding (boric acid dynamic bonding), systematically covering the entire protection chain from raw materials to finished product. This disruptive solution addresses three key pain points: retention rate increases from ≤60% to ≥90%; 2-hour release rate decreases from >80% to <30%; and precipitation decreases from >20% to ≤4%.

[0024] Simultaneously, intelligent responsive release is achieved: the pH responsiveness of the borate ester bond enables adaptive sustained release in the wound environment. Through a three-level synergistic mechanism of molecular pre-encapsulation → physical barrier → chemical bonding, this invention fundamentally overcomes the technical barriers to the industrialization of tea polyphenol dressings, providing research direction for the development of high-end functional dressings with long-lasting stability and intelligent response. DETAILED DESCRIPTION

[0025] Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, it is obvious that described embodiment is only a part of embodiment of the present invention, rather than whole embodiment. Based on the embodiment in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work premise all fall within the scope of protection of the present invention. Meanwhile, raw materials, reagents or devices used in the following examples, unless otherwise specified, can be obtained from conventional commercial channels, or can be obtained by existing known methods.

[0026] Example 1

[0027] Preparation of Oriental Beauty Tea Extract: Commercially available Fujian Datian Oriental Beauty Tea was dried at room temperature, crushed and passed through a 65-mesh sieve to obtain tea powder, which was stored in the dark. An extract was added to the tea powder, wherein the extract was a 20% ethanol solution by volume, and cellulase (purchased from Sinopharm Group) and hydroxypropyl-β-cyclodextrin (purchased from Sinopharm Group) were added thereto. After the addition was completed, the system was heated to 45°C and extracted at a constant temperature for 20 minutes. After completion, the system was centrifuged at 4500 r / min for 10 minutes, and the supernatant was rotary evaporated and freeze-dried to a powder for later use to obtain the Oriental Beauty Tea extract, wherein the amount ratio of tea powder, extract, cellulase and hydroxypropyl-β-cyclodextrin was 2g:10mL:0.05g:0.1g.

[0028] Example 2

[0029] Preparation of Oriental Beauty Tea Extract: The commercially available Fujian Datian Oriental Beauty tea was oven-dried at room temperature, crushed and passed through a 65-mesh sieve to obtain tea powder, which was then stored in the dark. An extraction solution was added to the tea powder, the extraction solution being 50% ethanol by volume, and cellulase (purchased from the National Pharmaceutical Group) and hydroxypropyl-beta-cyclodextrin (purchased from the National Pharmaceutical Group) were then added. After the addition was complete, the system was warmed to 50°C, and constant temperature extraction was performed for 60 minutes. After completion, centrifugation was performed at 4500 r / min for 10 minutes, and the supernatant was rotary evaporated and freeze-dried into powder for later use to obtain the Oriental Beauty tea extract. The amount ratio of the tea powder, the extraction solution, the cellulase and the hydroxypropyl-beta-cyclodextrin was 2g:10mL:0.20g:1.0g.

[0030] Example 3

[0031] Preparation of the Oriental Beauty tea extract: The commercially available Fujian Datian Oriental Beauty tea was oven-dried at room temperature, crushed and passed through a 65-mesh sieve to obtain tea powder, which was then stored in the dark. An extraction solution was added to the tea powder, the extraction solution being 50% ethanol by volume, and cellulase (purchased from the National Pharmaceutical Group) and hydroxypropyl-beta-cyclodextrin (purchased from the National Pharmaceutical Group) were then added. After the addition was complete, the system was warmed to 50°C, and constant temperature extraction was performed for 60 minutes. After completion, centrifugation was performed at 4500 r / min for 10 minutes, and the supernatant was rotary evaporated and freeze-dried into powder for later use to obtain the Oriental Beauty tea extract. The amount ratio of the tea powder, the extraction solution, the cellulase and the hydroxypropyl-beta-cyclodextrin was 2g:10mL:0.20g:1.0g.

[0032] Comparative Example 1 Comparative Example 1 is a control group of Example 2, and the hydroxypropyl-beta-cyclodextrin in Example 2 is omitted, and the remaining parameters remain the same as in Example 2 to obtain the Oriental Beauty tea extract. The specific steps are as follows: Preparation of the Oriental Beauty tea extract: The commercially available Fujian Datian Oriental Beauty tea was oven-dried at room temperature, crushed and passed through a 65-mesh sieve to obtain tea powder, which was then stored in the dark. An extraction solution was added to the tea powder, the extraction solution being 50% ethanol by volume, and cellulase (purchased from the National Pharmaceutical Group) and hydroxypropyl-beta-cyclodextrin (purchased from the National Pharmaceutical Group) were then added. After the addition was complete, the system was warmed to 50°C, and constant temperature extraction was performed for 60 minutes. After completion, centrifugation was performed at 4500 r / min for 10 minutes, and the supernatant was rotary evaporated and freeze-dried into powder for later use to obtain the Oriental Beauty tea extract. The amount ratio of the tea powder, the extraction solution, the cellulase and the hydroxypropyl-beta-cyclodextrin was 2g:10mL:0.20g:1.0g.

[0033] Comparative Example 2 Comparative Example 2 is the control group of Example 2, omitting the cellulase and hydroxypropyl-β-cyclodextrin in Example 2, and extracting the tea powder with only a 50% ethanol solution by volume. The other parameters remain the same as in Example 2, and the Oriental Beauty Tea extract is obtained. The specific steps are as follows: Preparation of Oriental Beauty Tea Extract: Commercially available Fujian Datian Oriental Beauty tea was dried at room temperature, crushed and passed through a 65-mesh sieve to obtain tea powder, which was then stored in the dark. An extract was added to the tea powder, wherein the extract was a 50% ethanol solution by volume. The system was heated to 50°C and subjected to constant temperature extraction for 60 minutes. After completion, the extract was centrifuged at 4500 rpm for 10 minutes. The supernatant was rotary evaporated and freeze-dried to a powder for later use to obtain the Oriental Beauty tea extract, wherein the ratio of tea powder to extract was 2 g:10 mL.

[0034] Example 4

[0035] The Oriental Beauty Tea extract prepared in Example 1 was applied to the dressing: S1. Dissolve β-cyclodextrin (purchased from Sinopharm Group) in an ethanol solution at 58°C to obtain a β-cyclodextrin mixed solution, wherein the ethanol solution is composed of pure water and anhydrous ethanol in a volume ratio of 2:1, and then dissolve the Oriental Beauty Tea extract prepared in Example 1 in anhydrous ethanol to obtain an extract mixture. After the β-cyclodextrin mixed solution is cooled to 45°C, the extract mixture is added to the β-cyclodextrin mixed solution to obtain a mixed solution. The mixed solution is placed in the dark and stirred at a constant temperature at 200r / min for 10min. After completion, the mixed solution is placed in a low temperature environment of 4°C and allowed to stand overnight to precipitate crystals. The solid component is collected by filtration, and the solid component is washed with anhydrous ethanol and vacuum dried to finally obtain a β-cyclodextrin inclusion complex, wherein the amount ratio of β-cyclodextrin, ethanol solution, Oriental Beauty Tea extract prepared in Example 1, and anhydrous ethanol is 5g:100mL:2g:40mL.

[0036] S2. Sodium alginate (98%, purchased from Yuanye Bio) and 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) were dissolved in pure water, and carbodiimide hydrochloride (purchased from Hangzhou Xiaoyou Biotechnology Co., Ltd.) was added thereto with stirring to obtain a mixed solution. The mixed solution was stirred at room temperature for 24 hours. After completion, it was dialyzed with a dialysis bag with a molecular weight cutoff of 8000-14000Da (purchased from Shanghai Jingke Chemical Technology Co., Ltd., dialysis bag MD10) for 3 days. Finally, after freeze-drying, modified sodium alginate was obtained, wherein the amount ratio of sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride was 10g:3.9g:1L:9.6g.

[0037] S3, dissolve hyaluronic acid (purchased from source leaf biological) in a mixed solvent, then stir 3-aminobenzene boronic acid monohydrate (purchased from aladdin reagent) and 4-(4, 6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride (97%, purchased from macclin biological) into the mixed solvent, then stir 1M sodium hydroxide solution into the system to adjust the pH to 6.4, after completion, the system is stirred at room temperature for 48h, after completion, the system is dialyzed for 4 days with a dialysis bag with a molecular weight cutoff of 8000-14000Da (purchased from Shanghai jierke chemical technology co., LTD, dialysis bag MD10), the solid components are collected and freeze-dried to obtain modified hyaluronic acid, wherein the mixed solvent is prepared by mixing pure water and dimethylformamide in a volume ratio of 3:2, and the amount of hyaluronic acid, mixed solvent, 3-aminobenzene boronic acid monohydrate, 4-(4, 6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride is 5g:100ml:1.56g:3.46g.

[0038] S4, stir modified modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion compound into pure water to obtain a mixed solution, then stir 1.0M sodium hydroxide solution into it, stir at room temperature for 3min, then stand for 30min, after completion, sterilize to obtain a gel-like dressing, wherein the amount of pure water, modified modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0M sodium hydroxide solution is 200ml:12g:15g:3g:4g:10ml.

[0039] Example 5

[0040] The oriental beauty tea extract prepared in example 2 is applied to the dressing: S1, dissolve β-cyclodextrin (purchased from China national pharmaceutical group) in 60℃ ethanol solution to obtain β-cyclodextrin mixed solution, wherein the ethanol solution is prepared by mixing pure water and anhydrous ethanol in a volume ratio of 2:1, then dissolve the oriental beauty tea extract prepared in the above example 2 in anhydrous ethanol to obtain extract mixture, after the β-cyclodextrin mixed solution is cooled to 50℃, add the extract mixture into the β-cyclodextrin mixed solution to obtain a mixed solution, place the mixed solution in the dark and stir at a constant temperature of 200r / min for 30min, after completion, place the mixed solution in a 4℃ low temperature environment and stand overnight, precipitate the crystals, filter and collect the solid components, then wash the solid components with anhydrous ethanol and vacuum dry to finally obtain β-cyclodextrin inclusion compound, wherein the amount of β-cyclodextrin, ethanol solution, oriental beauty tea extract prepared in example 2, anhydrous ethanol is 5g:100ml:2g:50ml.

[0041] S2. Sodium alginate (98%, purchased from Yuanye Bio) and 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) were dissolved in pure water, and carbodiimide hydrochloride (purchased from Hangzhou Xiaoyou Biotechnology Co., Ltd.) was added thereto with stirring to obtain a mixed solution. The mixed solution was stirred at room temperature for 24 hours. After completion, it was dialyzed with a dialysis bag with a molecular weight cutoff of 8000-10000Da (purchased from Nantong Feiyu Biotechnology Co., Ltd.) for 3 days. Finally, after freeze-drying, modified sodium alginate was obtained, wherein the amount ratio of sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride was 10g:4.0g:1L:10.0g.

[0042] S3. Dissolve hyaluronic acid (purchased from Yuanye Bio) in a mixed solvent, then add 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (97%, purchased from MacLean Bio) with stirring, then add 1M sodium hydroxide solution with stirring to adjust the pH of the system to 6.5. After completion, the system is stirred and reacted at room temperature for 48 hours. After completion, use a 5-well plate with a molecular weight cutoff of 8000- The product was dialyzed for 4 days using a 10,000Da dialysis bag (purchased from Nantong Feiyu Biotechnology Co., Ltd.), and the solid components were collected and freeze-dried to obtain modified hyaluronic acid. The mixed solvent was prepared by mixing pure water and dimethylformamide in a volume ratio of 3:2, and the dosage ratio of hyaluronic acid, mixed solvent, 3-aminophenylboronic acid monohydrate, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride was 5 g:100 mL:1.60 g:3.50 g.

[0043] S4. Add modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion compound to pure water with stirring to obtain a mixed solution, and then add 1.0 M sodium hydroxide solution thereto with stirring, stir at room temperature for 5 minutes, and then let it stand for 30 minutes. After completion, sterilize to obtain a gel dressing, wherein the amount ratio of pure water, modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0 M sodium hydroxide solution is 200 mL:14 g:14 g:2 g:5 g:10 mL.

[0044] Example 6

[0045] The Oriental Beauty Tea extract prepared in Example 3 was applied to the dressing: S1. Dissolve β-cyclodextrin (purchased from Sinopharm Group) in an ethanol solution at 60°C to obtain a β-cyclodextrin mixed solution, wherein the ethanol solution is composed of pure water and anhydrous ethanol in a volume ratio of 2:1, and then dissolve the Oriental Beauty Tea extract prepared in Example 3 in anhydrous ethanol to obtain an extract mixture. After the β-cyclodextrin mixed solution is cooled to 50°C, the extract mixture is added to the β-cyclodextrin mixed solution to obtain a mixed solution. The mixed solution is placed in the dark and stirred at a constant temperature at 200r / min for 30min. After completion, the mixed solution is placed in a low temperature environment of 4°C and allowed to stand overnight to precipitate crystals. The solid component is collected by filtration, and the solid component is washed with anhydrous ethanol and vacuum dried to finally obtain a β-cyclodextrin inclusion complex, wherein the amount ratio of β-cyclodextrin, ethanol solution, Oriental Beauty Tea extract prepared in Example 3, and anhydrous ethanol is 5g:100mL:2g:50mL.

[0046] S2. Sodium alginate (98%, purchased from Yuanye Bio) and 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) were dissolved in pure water, and carbodiimide hydrochloride (purchased from Hangzhou Xiaoyou Biotechnology Co., Ltd.) was added thereto with stirring to obtain a mixed solution. The mixed solution was stirred at room temperature for 24 hours. After completion, it was dialyzed with a dialysis bag with a molecular weight cutoff of 8000-14000Da (purchased from Shanghai Jingke Chemical Technology Co., Ltd., dialysis bag MD10) for 3 days. Finally, after freeze-drying, modified sodium alginate was obtained, wherein the amount ratio of sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride was 10g:4.0g:1L:10.0g.

[0047] S3, dissolving hyaluronic acid (purchased from Yuanye Bio) in a mixed solvent, then adding 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (97%, purchased from MacLean Bio) with stirring, then adding 1M sodium hydroxide solution with stirring to adjust the pH of the system to 6.8, after completion, the system was stirred at room temperature for 48 hours, after completion, using a cut-off molecular weight of 8000-1400 The hyaluronic acid was dialyzed for 4 days using a dialysis bag with a diameter of 0 Da (purchased from Shanghai Jingke Chemical Technology Co., Ltd., dialysis bag MD10), and the solid components were collected and freeze-dried to obtain modified hyaluronic acid, wherein the mixed solvent was prepared by pure water and dimethylformamide in a volume ratio of 3:2, and the amount ratio of hyaluronic acid, mixed solvent, 3-aminophenylboronic acid monohydrate, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride was 5 g:100 mL:1.60 g:3.50 g.

[0048] S4. Add modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion compound to pure water with stirring to obtain a mixed solution, and then add 1.0 M sodium hydroxide solution thereto with stirring, stir at room temperature for 5 minutes, and then let it stand for 30 minutes. After completion, sterilize to obtain a gel dressing, wherein the amount ratio of pure water, modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0 M sodium hydroxide solution is 200 mL: 15 g: 12 g: 3 g: 5 g: 10 mL.

[0049] Comparative Example 3 Comparative Example 3 is the control group of Example 5, in which the Oriental Beauty Tea extract prepared in Example 2 of Example 5 is replaced by the Oriental Beauty Tea extract prepared in Comparative Example 1, and the remaining raw materials, raw material amounts and preparation steps remain the same as in Example 5, and a gel dressing is finally obtained.

[0050] Comparative Example 4 Comparative Example 4 is the control group of Example 5. The Oriental Beauty Tea extract prepared in Example 2 in Example 5 is replaced by the Oriental Beauty Tea extract prepared in Comparative Example 2. The remaining raw materials, raw material amounts and preparation steps remain the same as in Example 5, and a gel dressing is finally obtained.

[0051] Comparative Example 5 Comparative Example 5 is the control group of Example 5. Step S1 in Example 5 is removed, that is, the β-cyclodextrin inclusion step (S1) is omitted, and the Oriental Beauty Tea extract is directly added to the dressing base material. The remaining raw materials, raw material amounts and preparation steps remain the same as in Example 5, and a gel dressing is finally obtained. The specific steps are as follows: The Oriental Beauty Tea extract prepared in Example 2 was applied to the dressing: S1. Sodium alginate (98%, purchased from Yuanye Bio) and 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) were dissolved in pure water, and carbodiimide hydrochloride (purchased from Hangzhou Xiaoyou Biotechnology Co., Ltd.) was added thereto with stirring to obtain a mixed solution. The mixed solution was stirred at room temperature for 24 hours. After completion, it was dialyzed with a dialysis bag with a molecular weight cutoff of 8000-10000Da (purchased from Nantong Feiyu Biotechnology Co., Ltd.) for 3 days. Finally, after freeze-drying, modified sodium alginate was obtained, wherein the amount ratio of sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride was 10g:4.0g:1L:10.0g.

[0052] S2. Dissolve hyaluronic acid (purchased from Yuanye Bio) in a mixed solvent, then add 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (97%, purchased from MacLean Bio) with stirring, then add 1M sodium hydroxide solution with stirring to adjust the pH of the system to 6.5. After completion, the system is stirred and reacted at room temperature for 48 hours. After completion, use a 5-well plate with a molecular weight cutoff of 8000 to 100 μg / ml. The product was dialyzed for 4 days using a 10,000Da dialysis bag (purchased from Nantong Feiyu Biotechnology Co., Ltd.), and the solid components were collected and freeze-dried to obtain modified hyaluronic acid. The mixed solvent was prepared by mixing pure water and dimethylformamide in a volume ratio of 3:2, and the dosage ratio of hyaluronic acid, mixed solvent, 3-aminophenylboronic acid monohydrate, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride was 5 g:100 mL:1.60 g:3.50 g.

[0053] S3. Add modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and the Oriental Beauty Tea extract prepared in Example 2 to pure water with stirring to obtain a mixed solution, and then add 1.0 M sodium hydroxide solution thereto with stirring, stir at room temperature for 5 minutes, and then let it stand for 30 minutes. After completion, sterilize to obtain a gel dressing, wherein the amount ratio of pure water, modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, the Oriental Beauty Tea extract prepared in Example 2 and 1.0 M sodium hydroxide solution is 200 mL: 14 g: 14 g: 2 g: 5 g: 10 mL.

[0054] Comparative Example 6 Comparative Example 6 is the control group of Example 5. Step S2 in Example 5 is removed, that is, unmodified sodium alginate (98%, purchased from Yuanye Bio) is used for subsequent dressing preparation. The remaining raw materials, raw material amounts and preparation steps are kept consistent with those in Example 5, and a gel dressing is finally obtained. The specific steps are as follows: The Oriental Beauty Tea extract prepared in Example 2 was applied to the dressing: S1, dissolve β-cyclodextrin (purchased from the country drug group) in 60℃ ethanol solution to obtain β-cyclodextrin mixed solution, wherein the ethanol solution is prepared by pure water and anhydrous ethanol according to the volume ratio of 2:1, then dissolve the extract of oriental beauty tea prepared in the above example 2 in anhydrous ethanol to obtain extract mixed solution, after the β-cyclodextrin mixed solution is cooled to 50℃, the extract mixed solution is added to the β-cyclodextrin mixed solution to obtain the mixed solution, the mixed solution is placed in the dark condition and stirred at 200r / min for 30min, then the mixed solution is placed in 4℃ low temperature environment for overnight, the crystal is precipitated, the solid component is collected by filtration, the solid component is washed by anhydrous ethanol and vacuum dried, finally the β-cyclodextrin inclusion compound is obtained, wherein the amount ratio of β-cyclodextrin, ethanol solution, oriental beauty tea extract prepared in example 2, anhydrous ethanol is 5g:100mL:2g:50mL.

[0055] S2, dissolve hyaluronic acid (purchased from source leaf biological) in mixed solvent, then add 3-aminobenzene boronic acid monohydrate (purchased from aladdin reagent) and 4-(4,6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride (97%, purchased from macclin biological) into the stirring system, then add 1M sodium hydroxide solution to adjust the pH of the system to 6.5, after completion, the system is stirred at room temperature for 48h, then the system is dialyzed for 4 days by using dialysis bag with molecular weight cutoff of 8000-10000Da (purchased from nantong feiyu biological technology co., ltd.), the solid component is collected and freeze-dried to obtain modified hyaluronic acid, wherein the mixed solvent is prepared by pure water and dimethylformamide according to the volume ratio of 3:2, wherein the amount ratio of hyaluronic acid, mixed solvent, 3-aminobenzene boronic acid monohydrate and 4-(4,6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride is 5g:100mL:1.60g:3.50g.

[0056] S3, add sodium alginate (98%, purchased from source leaf biological), modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion compound into pure water to obtain mixed solution, then add 1.0M sodium hydroxide solution into the stirring system, stir at room temperature for 5min, then stand for 30min, after completion, sterilize to obtain gel-like dressing, wherein the amount ratio of pure water, sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0M sodium hydroxide solution is 200mL:14g:14g:2g:5g:10mL.

[0057] Comparative example 7 Comparative Example 7 is the control group of Example 5. Step S3 in Example 5 is removed, that is, unmodified hyaluronic acid (purchased from Yuanye Bio) is used for subsequent dressing preparation. The remaining raw materials, raw material amounts and preparation steps remain the same as in Example 5, and a gel dressing is finally obtained. The specific steps are as follows: The Oriental Beauty Tea extract prepared in Example 2 was applied to the dressing: S1. Dissolve β-cyclodextrin (purchased from Sinopharm Group) in an ethanol solution at 60°C to obtain a β-cyclodextrin mixed solution, wherein the ethanol solution is composed of pure water and anhydrous ethanol in a volume ratio of 2:1, and then dissolve the Oriental Beauty Tea extract prepared in Example 2 in anhydrous ethanol to obtain an extract mixture. After the β-cyclodextrin mixed solution is cooled to 50°C, the extract mixture is added to the β-cyclodextrin mixed solution to obtain a mixed solution. The mixed solution is placed in the dark and stirred at a constant temperature at 200r / min for 30min. After completion, the mixed solution is placed in a low temperature environment of 4°C and allowed to stand overnight to precipitate crystals. The solid component is collected by filtration, and the solid component is washed with anhydrous ethanol and vacuum dried to finally obtain a β-cyclodextrin inclusion complex, wherein the amount ratio of β-cyclodextrin, ethanol solution, Oriental Beauty Tea extract prepared in Example 2, and anhydrous ethanol is 5g:100mL:2g:50mL.

[0058] S2. Sodium alginate (98%, purchased from Yuanye Bio) and 3-aminophenylboronic acid monohydrate (purchased from Aladdin Reagent) were dissolved in pure water, and carbodiimide hydrochloride (purchased from Hangzhou Xiaoyou Biotechnology Co., Ltd.) was added thereto with stirring to obtain a mixed solution. The mixed solution was stirred at room temperature for 24 hours. After completion, it was dialyzed with a dialysis bag with a molecular weight cutoff of 8000-10000Da (purchased from Nantong Feiyu Biotechnology Co., Ltd.) for 3 days. Finally, after freeze-drying, modified sodium alginate was obtained, wherein the amount ratio of sodium alginate, 3-aminophenylboronic acid monohydrate, pure water, and carbodiimide hydrochloride was 10g:4.0g:1L:10.0g.

[0059] S3. Modified sodium alginate, hyaluronic acid (purchased from Source Leaf Bio), polyvinyl alcohol and β-cyclodextrin inclusion compound were added to pure water with stirring to obtain a mixed solution, and 1.0 M sodium hydroxide solution was added thereto with stirring. The mixture was stirred at room temperature for 5 min, and allowed to stand for 30 min. After completion, the mixture was sterilized to obtain a gel dressing, wherein the amount ratio of pure water, modified sodium alginate, hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0 M sodium hydroxide solution was 200 mL: 14 g: 14 g: 2 g: 5 g: 10 mL.

[0060] Test Example 1 The dressings prepared in Examples 4 to 6 and Comparative Examples 3 to 7 were subjected to performance tests. The performance test process is as follows, and the test results are shown in Table 1: (1) Active ingredient stability test: Test process: Each dressing sample (Examples 4 to 6, Comparative Examples 3 to 7) was cut into 1 cm × 1 cm slices (mass of about 0.5 g) and placed in an autoclave for high-temperature sterilization (121 ° C, 15 min, simulating the sterilization process). After sterilization, the sample was stored at 25 ° C in the dark for 30 days (simulating long-term storage). After storage, 0.5 g of the sample was taken, 10 mL of 70% ethanol solution was added, ultrasonic extraction was performed at 40 ° C for 30 min, centrifuged (4500 r / min, 10 min), and the supernatant was taken. The polyphenol content was measured by the Folin-Ciocalteu method: 0.5 mL of the supernatant was taken, 2.5 mL of Folin reagent (diluted 10 times), shaken and allowed to stand for 5 min, and then 2 mL of 7.5% Na2CO3 solution was added, the volume was made up to 10 mL, and the reaction was carried out in the dark at room temperature for 1 h. The absorbance was measured at 765 nm, and the polyphenol concentration was calculated using gallic acid as the standard.

[0061] Calculate the stability retention rate: Retention rate (%) = (polyphenol content after storage / initial polyphenol content) × 100%, where the initial polyphenol content is measured immediately after preparation.

[0062] (2) Sustained release effect test: Testing process: 0.5 g of each dressing sample (Examples 4-6, Comparative Examples 3-7) was placed in 50 mL of PBS buffer (pH 7.4, 37°C) and placed in a constant temperature shaker (37°C, 100 rpm). At 0 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 12 h, 1 mL of sample was taken (1 mL of fresh PBS was added at the same time). The supernatant was centrifuged and the absorbance of the released solution at 280 nm (polyphenol characteristic peak) was measured by ultraviolet spectrophotometry. The cumulative release percentage was calculated using the standard curve.

[0063] Calculate key indicators: 2-hour and 12-hour cumulative release percentages.

[0064] (3) Compatibility test: Test process: Prepare dressing samples (Examples 4-6, Comparative Examples 3-7) and pour them into transparent test tubes. Let them stand at room temperature for 48 hours, then centrifuge them (3000 rpm, 10 minutes), collect the precipitate, and weigh it.

[0065] Calculate the percentage of sedimentation: sedimentation (%) = (precipitation weight / total dressing weight) × 100%.

[0066]

[0067] According to the data analysis in Table 1: (1) Active ingredient stability (polyphenol retention rate): Examples 4-6 (retention rate 89.8%-93.2%): using three-step protection chain technology: A. Hydroxypropyl-β-cyclodextrin (HP-β-CD) was added during the extraction phase to protect polyphenols; B. β-cyclodextrin inclusion (encapsulation and isolation of oxygen / moisture); C. Borated substrate (dynamic covalent bonds reduce oxidation).

[0068] The synergistic effect significantly reduces polyphenol degradation during sterilization and storage.

[0069] Comparative Example 3 (64.7% retention): The extract from Comparative Example 1 was used (HP-β-CD omitted), retaining only the β-cyclodextrin inclusion complex and the borated matrix. The lack of protection during the extraction phase resulted in insufficient initial polyphenol stability, resulting in a nearly 30% decrease in retention.

[0070] Comparative Example 4 (retention rate 60.2%): The extract of Comparative Example 2 was used (without HP-β-CD and cellulase). The active ingredients in the base extract were easily oxidized, and subsequent protection could not completely compensate for this, resulting in the lowest retention rate.

[0071] Comparative Examples 5-7 (retention rates 52.3%-59.6%): Comparative Example 5: β-cyclodextrin inclusion was omitted, and the extract was added directly to the dressing; Comparative Examples 6 / 7: Borated sodium alginate / hyaluronic acid were omitted, respectively. The omission of any one of the protection steps resulted in a significant decrease in stability, demonstrating the essential importance of all three steps in the protection chain.

[0072] (2) Sustained release effect (2h / 12h cumulative release rate): Examples 4 to 6 (28.5% to 35.3% release in 2 hours; 74.9% to 80.2% release in 12 hours): β-cyclodextrin inclusion complex forms a "hydrophobic cavity encapsulation" and combines the pH-responsive bonding of the boronic acid group to achieve a dual-stage sustained-release mechanism.

[0073] Comparative Example 3 (41.2% release in 2 h; 92.3% release in 12 h): Using an extract not protected by HP-β-CD, the sustained-release effect after inclusion was weaker than that in Example 3 (12% higher release rate in 12 h), proving that protection during the extraction stage affects inclusion stability.

[0074] Comparative Example 4 (85.7% released in 2 hours; 99.1% released in 12 hours): The active ingredient of the crude extract without enzyme-assisted extraction was easily released, and the sustained-release mechanism failed.

[0075] Comparative Example 5 (2h release 88.6%; 12h release 99.5%): Direct addition of extract (without inclusion), active ingredient released rapidly, 2h release rate over 85%, proving inclusion is the sustained release core.

[0076] Comparative Examples 6-7 (2h release 49.8-51.4%; 12h release 87.9-89.2%): Without boronated substrate, inclusion provides some sustained release, but lack of dynamic covalent bond results in release rate still significantly higher than examples (12h release rate about 10% higher).

[0077] (3) Compatibility (percentage of precipitate): Examples 4-6 (precipitate 2.1-4.0%): Dynamic covalent bond between boronate groups of boronated sodium alginate / hyaluronic acid and o-diphenol hydroxyl groups of polyphenols effectively inhibits phase separation.

[0078] Comparative Examples 3-4 (precipitate 16.4-24.9%): Poor stability of extract itself (lack of HP-β-CD), exacerbates incompatibility with substrate, precipitate significantly increased.

[0079] Comparative Example 5 (precipitate 26.3%): Polyphenols not included, directly exposed, interact with substrate resulting in severe precipitation.

[0080] Comparative Examples 6-7 (precipitate 19.8-21.7%): Without boronated modification (Comparative Example 6 lacks sodium alginate modification; Comparative Example 7 lacks hyaluronic acid modification), dynamic bonding mechanism is destroyed, precipitate increased by more than 5 times compared to examples.

[0081] It should be noted that in this document, such as the term "comprising, including" or any other variant thereof is intended to cover non-exclusive inclusion, so that the process, method, article or device including a series of elements not only includes those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0082] Although embodiments of the present application have been shown and described, it is to be understood that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present application, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An Oriental Beauty Tea extract, characterized in that Prepared by the following steps: Oriental Beauty tea is dried and pulverized through a 65-mesh sieve to obtain tea powder, an extract is added to the tea powder, and then cellulase and hydroxypropyl-β-cyclodextrin are added thereto. The system is heated to 45-50°C and subjected to constant temperature extraction for 20-60 minutes. After extraction, the system is centrifuged at 4500 rpm for 10 minutes, the supernatant is rotary evaporated, and then freeze-dried to a powder to obtain the Oriental Beauty tea extract.

2. The Oriental Beauty Tea extract according to claim 1, characterized in that The extract is an ethanol solution with a volume fraction of 20% to 80%.

3. The Oriental Beauty Tea extract according to claim 1, characterized in that The usage ratio of the tea powder, the extract, the cellulase and the hydroxypropyl-β-cyclodextrin is 2g:10mL:0.05-0.25g:0.1-1.0g.

4. The use of an Oriental Beauty Tea extract in a medical dressing according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1. Dissolve β-cyclodextrin in an ethanol solution at 58-60° C. to obtain a β-cyclodextrin mixed solution, dissolve the Oriental Beauty Tea extract in anhydrous ethanol to obtain an extract mixed solution, and after the β-cyclodextrin mixed solution is cooled to 45-50° C., add the extract mixed solution to the β-cyclodextrin mixed solution to obtain a mixed solution, place the mixed solution in the dark, and stir at a constant temperature for 10-30 minutes. After completion, place the mixed solution in a low-temperature environment at 4° C. and let it stand overnight to precipitate crystals, collect the solid component by filtration, and then wash the solid component with anhydrous ethanol and vacuum dry to obtain a β-cyclodextrin inclusion compound; S2. Add modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol and β-cyclodextrin inclusion complex into pure water with stirring to obtain a mixed solution, then add 1.0 M sodium hydroxide solution thereto with stirring, stir at room temperature for 3 to 5 minutes, let stand for 30 minutes, and sterilize after completion to obtain a gel dressing.

5. The use according to claim 4, characterized in that The ethanol solution in S1 is prepared by mixing pure water and anhydrous ethanol in a volume ratio of 2:1; the dosage ratio of the β-cyclodextrin, ethanol solution, Oriental Beauty Tea extract, and anhydrous ethanol is 5g:100mL:2g:40-50mL.

6. The use according to claim 4, characterized in that The usage ratio of pure water, modified sodium alginate, modified hyaluronic acid, polyvinyl alcohol, β-cyclodextrin inclusion compound and 1.0M sodium hydroxide solution in S2 is 200mL:12~15g:12~15g:2~3g:4~5g:10mL.

7. The use according to claim 4, characterized in that The modified sodium alginate described in S2 is prepared by the following steps: Sodium alginate and 3-aminophenylboronic acid monohydrate are dissolved in pure water, and then carbodiimide hydrochloride is added thereto with stirring to obtain a mixed solution. The mixed solution is stirred at room temperature for 24 hours. After completion, it is dialyzed using a dialysis bag with a molecular weight cutoff of 8000 to 14000 Da for 3 days, and finally freeze-dried to obtain modified sodium alginate.

8. The use according to claim 7, characterized in that The usage ratio of the sodium alginate, 3-aminophenylboronic acid monohydrate, pure water and carbodiimide hydrochloride is 10g:3.9-4.0g:1L:9.6-10.0g.

9. The use according to claim 4, characterized in that The modified hyaluronic acid described in S2 is prepared by the following steps: Hyaluronic acid was dissolved in a mixed solvent, and 3-aminophenylboronic acid monohydrate and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride were added thereto with stirring. 1M sodium hydroxide solution was then added thereto with stirring to adjust the pH of the system to 6.4-6.

8. After completion, the system was stirred and reacted at room temperature for 48 hours. After completion, the system was dialyzed for 4 days using a dialysis bag with a molecular weight cutoff of 8000-14000 Da, and the solid component was collected and freeze-dried to obtain modified hyaluronic acid.

10. The use according to claim 9, characterized in that The mixed solvent is prepared by pure water and dimethylformamide in a volume ratio of 3:2; the usage ratio of the hyaluronic acid, the mixed solvent, 3-aminophenylboronic acid monohydrate, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride is 5g:100mL:1.56~1.60g:3.46~3.50g.

Images