Composition for treating intractable skin infection, sores and ulcers and inflammation as well as preparation method and application of composition

Abstract

The invention discloses a composition for treating intractable skin infection, sores and ulcers and inflammation as well as a preparation method and application of the composition, and belongs to the technical field of functional compositions. The composition is prepared from the following components in parts by mass: 1 to 5 parts of baicalin, 0.5 to 3 parts of antibacterial alkaloid, 2 to 6 parts of asiaticoside, 3 to 8 parts of salvianolic acid B, 1 to 4 parts of gallic acid, 5 to 10 parts of rhizoma bletillae polysaccharide, 3 to 10 parts of konjac glucomannan, 3 to 5 parts of chitosan, 8 to 20 parts of glycerol, 0.5 to 2 parts of genipin, 1 to 3 parts of calcium chloride, 1 to 3 parts of laurocapram and 40 to 80 parts of water. The antibacterial alkaloid comprises berberine or phellodendrine. Based on deep understanding of an intractable skin pathological mechanism, the components are scientifically matched, and the composition shows a remarkable synergistic effect in in-vitro antibacterial, anti-biofilm and anti-inflammatory experiments. The preparation process is simple, low in cost and suitable for industrial production.

Description

Technical Field

[0001] This application belongs to the field of functional composition technology, and particularly relates to a composition for treating intractable skin infections, sores and inflammation, its preparation method and application. Background Technology

[0002] Refractory skin infections (such as folliculitis and boils caused by drug-resistant bacteria), chronic sores (such as diabetic foot ulcers and pressure sores), and severe skin inflammations (such as severe eczema and dermatitis) pose significant challenges to clinical treatment. Their common pathological features include severely impaired wound barrier function, bacterial biofilm formation, persistent excessive inflammatory response, and low tissue repair and regeneration capacity. Current mainstream treatments, such as antibiotics (e.g., mupirocin ointment) and corticosteroids, often exhibit poor efficacy, drug resistance, high recurrence rates, and significant side effects when dealing with biofilm-related infections and chronic inflammation.

[0003] Baicalin, berberine, and other natural active ingredients have been proven to possess anti-inflammatory, antibacterial, antioxidant, and healing-promoting effects, respectively. High molecular weight materials such as Bletilla striata polysaccharides and konjac glucomannan are commonly used as pharmaceutical excipients. However, current technologies mostly involve the application of single components or simple physical mixtures, failing to address the compatibility, stability, and synergistic delivery issues of multiple components. For example, simply mixed ointments struggle to penetrate biological membrane barriers, and the rapid drug release fails to maintain a long-lasting effective concentration on the wound surface.

[0004] Therefore, developing a novel drug composition that can synergistically target multiple sites, effectively penetrate and clear biofilms, regulate the inflammatory microenvironment, and continuously promote tissue regeneration has significant clinical and market value. Summary of the Invention

[0005] To overcome the limitations of existing technologies in treating biofilm-associated infections and addressing the multiple pathological aspects of chronic wounds, this application provides a composition for treating refractory skin infections, sores, and inflammation, along with its preparation method and application.

[0006] To achieve the above-mentioned objectives, this application provides the following technical solution:

[0007] On the one hand, this application provides a composition for treating intractable skin infections, sores and inflammation, prepared from components comprising the following parts by weight: 1-5 parts baicalin, 0.5-3 parts antibacterial alkaloids, 2-6 parts asiaticoside, 3-8 parts salvianolic acid B, 1-4 parts gallic acid, 5-10 parts Bletilla striata polysaccharide, 3-10 parts konjac glucomannan, 3-5 parts chitosan, 8-20 parts glycerol, 0.5-2 parts genipin, 1-3 parts calcium chloride, 1-3 parts laurocapram, and 40-80 parts water;

[0008] The antibacterial alkaloids include berberine or phellodendron chinense.

[0009] Optionally, the composition for treating intractable skin infections, sores, and inflammation is prepared from components comprising the following parts by weight: 3 parts baicalin, 2 parts antibacterial alkaloids, 4 parts asiaticoside, 5 parts salvianolic acid B, 3 parts gallic acid, 7 parts Bletilla striata polysaccharide, 5 parts konjac glucomannan, 4 parts chitosan, 12 parts glycerol, 1 part genipin, 2 parts calcium chloride, 2 parts laurocapram, and 50 parts water.

[0010] The antibacterial alkaloids include berberine or phellodendron chinense.

[0011] Optionally, the degree of deacetylation of the chitosan is ≥85%, and the molecular weight of the chitosan is 5000-20000 Da.

[0012] Secondly, this application provides a method for preparing the above-mentioned composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0013] (1) Mix Bletilla striata polysaccharide, konjac glucomannan and water preheated to 55-60°C, stir and heat to 80-85°C, keep warm and stand, cool to obtain phase A;

[0014] (2) Mix chitosan with acidified water and stir until completely dissolved to obtain phase B;

[0015] (3) Baicalin, antibacterial alkaloids, asiaticoside, salvianolic acid B, gallic acid and glycerol were mixed and homogenized to obtain phase C;

[0016] (4) Mix laurocapram with glycerol to obtain phase D;

[0017] (5) Dissolve genipin and calcium chloride in water to obtain genipin solution and calcium chloride solution respectively;

[0018] (6) Add phase B to phase A and stir evenly. Then add phase C and continue stirring for 10-15 minutes. Then add genipin solution and react for 10-20 minutes. Finally, add calcium chloride solution to obtain a light yellow transparent gel.

[0019] (7) Add phase D to the pale yellow transparent gel, add the remaining water, stir evenly, and perform vacuum degassing to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0020] Optionally, in step (1), the stirring speed is 200-300 r / min;

[0021] The heat preservation and static setting time is 25-35 minutes;

[0022] The cooling process involves cooling the temperature to 25–40°C.

[0023] Optionally, in step (1), the stirring speed is independently selected from any value or a range between 200 r / min, 210 r / min, 220 r / min, 230 r / min, 240 r / min, 250 r / min, 260 r / min, 270 r / min, 280 r / min, 290 r / min, and 300 r / min.

[0024] Optionally, in step (1), the heat preservation time is independently selected from any value of 25min, 28min, 30min, 32min, 35min or any range between two.

[0025] Optionally, in step (1), the cooling is independently selected from any value of 25°C, 30°C, 35°C, 40°C, or a range between any two.

[0026] Optionally, in step (2), the pH of the acidified water is 4 to 5;

[0027] The acidification was carried out using glacial acetic acid;

[0028] The concentration of the glacial acetic acid is 1-3 wt%.

[0029] The stirring speed is 300-500 r / min.

[0030] Optionally, in step (2), the pH of the acidified water is independently selected from any value of 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5 or a range between any two.

[0031] Optionally, in step (2), the concentration of the glacial acetic acid is independently selected from any value of 1 wt%, 2 wt%, 3 wt%, or any range between two of them.

[0032] Optionally, in step (2), the stirring speed is independently selected from any value among 300 r / min, 350 r / min, 400 r / min, 450 r / min, and 500 r / min, or a range between any two.

[0033] Optionally, in step (3), the antibacterial alkaloids include berberine or phellodendron chinense;

[0034] The rotational speed of the homogenizer is 3000–4000 r / min;

[0035] The homogenization time is 5 to 10 minutes.

[0036] Optionally, in step (3), the homogeneous rotation speed is independently selected from any value among 3000 r / min, 3200 r / min, 3400 r / min, 3500 r / min, 3600 r / min, 3800 r / min, and 4000 r / min, or a range between any two.

[0037] Optionally, in step (3), the homogenization time is independently selected from any value of 5 min, 6 min, 7 min, 8 min, 9 min, 10 min or any range between two of them.

[0038] Optionally, the ratio of the amount of glycerol used in step (3) to the amount of glycerol used in step (4) is 1 to 3:1.

[0039] Optionally, in step (6), the stirring speed is 50 to 100 r / min.

[0040] Optionally, in step (6), the stirring speed is independently selected from any value among 50 r / min, 60 r / min, 70 r / min, 80 r / min, 90 r / min, and 100 r / min, or a range between any two.

[0041] Optionally, in step (7), the stirring speed is 300-500 r / min;

[0042] The vacuum degassing pressure is -0.1 to -0.05 MPa;

[0043] The vacuum degassing time is 10-15 minutes.

[0044] Optionally, in step (7), the stirring speed is independently selected from any value among 300 r / min, 350 r / min, 400 r / min, 450 r / min, and 500 r / min, or a range between any two.

[0045] Optionally, in step (7), the vacuum degassing pressure is independently selected from any value among -0.1 MPa, -0.09 MPa, -0.08 MPa, -0.07 MPa, -0.06 MPa, and -0.05 MPa, or a range between any two.

[0046] Optionally, in step (7), the vacuum degassing time is independently selected from any value of 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, or any range between the two.

[0047] Thirdly, this application provides the use of the above composition in the preparation of a medicament for treating intractable skin infections, sores, and inflammation.

[0048] Compared with the prior art, this application has the following advantages:

[0049] (1) The composition for treating intractable skin infections, sores, and inflammation provided in this application combines berberine / palmitin (which disrupts cell membranes / structures) with gallic acid (which inhibits enzyme activity), resulting in synergistic antibacterial and anti-biofilm effects; it combines baicalin (immunomodulatory), salvianolic acid B (antioxidant, improves microcirculation), and asiaticoside (stimulates collagen), achieving a seamless transition from inhibiting excessive and persistent inflammatory responses to actively promoting repair; and it incorporates Bletilla striata polysaccharide (a natural adhesive with film-forming and moisturizing properties), konjac glucomannan (which forms an irreversible gel upon contact with alkali), and low molecular weight chitosan (anti-inflammatory). Using a composite matrix of antibacterial, healing-promoting, and bioadhesive components, genipin and calcium chloride are synergistically cross-linked to form a stable hydrogel in situ on the skin wound. This gel can closely adhere to irregular wound surfaces, providing a long-lasting moist environment. At the same time, by cross-linking the polysaccharide chains, the release rate of active ingredients is regulated, achieving long-lasting sustained release, extending the duration of action, and improving efficacy. The added laurocapram, as a penetration enhancer, can effectively overcome skin barrier dysfunction and stratum corneum obstruction, ensuring that both hydrophobic active ingredients (such as baicalin and berberine) and hydrophilic ingredients can be efficiently absorbed transdermally, reaching deep into the lesion.

[0050] (2) The preparation method of the composition for treating intractable skin infections, sores, and inflammation provided in this application employs a unique stepwise in-situ crosslinking process. First, chitosan and other components are initially crosslinked with genipin (reacting with amino groups), then calcium chloride (ionically crosslinking with konjac glucomannan and other components) is introduced, ultimately forming a dense gel structure. This stepwise process precisely controls the gel formation process, ensuring that the active ingredients (especially hydrophobic components) are uniformly dispersed and encapsulated within the gel structure, guaranteeing the sustained-release properties and stability of the final product. Furthermore, the preparation process of this application is simple, low-cost, and suitable for industrial production. Detailed Implementation

[0051] The present application is further illustrated below with reference to specific embodiments. The following descriptions are merely a few embodiments of the present application and are not intended to limit the present application in any way. Although the present application discloses preferred embodiments as follows, they are not intended to limit the present application. Any modifications or variations made by those skilled in the art without departing from the scope of the technical solution of the present application using the disclosed technical content are equivalent to equivalent implementation cases and all fall within the scope of the technical solution.

[0052] Unless otherwise specified, the raw materials used in the embodiments of this application are all purchased commercially and used directly without any special treatment.

[0053] Unless otherwise specified, the analytical methods in the embodiments all adopt conventional instrument or equipment settings and conventional analytical methods.

[0054] In the following examples and comparative examples, baicalin was purchased from Wuhan Kangqiong Biomedical Technology Co., Ltd. (active ingredient content 90%); berberine, asiaticoside, and genipin were purchased from Shanghai Yuanye Biotechnology Co., Ltd. (purity: HPLC ≥ 98%); berberine and konjac glucomannan were purchased from Jiangsu Caiwei Biotechnology Co., Ltd. (purity: 99%); salvianolic acid B was purchased from Nanjing Bencao Yikang Biotechnology Co., Ltd. (CAS No.: 121521-90-2, purity ≥ 98%); gallic acid was purchased from Hubei Xinrunde Chemical Co., Ltd. (CAS ≥ 99%). No.: 149-91-7 (purity 99%); Bletilla striata polysaccharide was purchased from Xi'an Green Biotechnology Co., Ltd. (purity 98%); Chitosan was purchased from Wuhan Jiyesheng Chemical Co., Ltd. (molecular weight 5000 Da); Glycerin was purchased from Yikang Natural Fragrance Oil Refinery in Jishui County, Jiangxi Province (purity 99%); Calcium chloride was purchased from Puyang Kaisheng Chemical Co., Ltd. (purity 99%); Laurozone was purchased from Hubei Boshi Chemical Co., Ltd. (purity 99%); Matrine was purchased from Xi'an Bainiankang Biotechnology Co., Ltd.; Paeonol was purchased from Xi'an Miaoguo Biotechnology Co., Ltd.

[0055] Example 1

[0056] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0057] (1) Weigh the following raw materials in the indicated mass parts: 1 part baicalin, 0.5 part berberine, 2 parts asiaticoside, 3 parts salvianolic acid B, 1 part gallic acid, 5 parts Bletilla striata polysaccharide, 3 parts konjac glucomannan, 3 parts chitosan, 8 parts glycerol, 0.5 parts genipin, 1 part calcium chloride, 1 part laurocapram, and 71 parts water.

[0058] (2) Mix Bletilla striata polysaccharide, konjac glucomannan and 25 parts of water preheated to 55°C, stir at 200 r / min and heat to 80°C, keep warm and stand for 25 min, cool to 25°C to obtain phase A;

[0059] (3) Chitosan is mixed with 10 parts of acidified water (pH 4, the acidification is carried out with 1 wt% glacial acetic acid) and stirred at 300 r / min until completely dissolved to obtain phase B;

[0060] (4) Baicalin, berberine, asiaticoside, salvianolic acid B, gallic acid and 6 parts of glycerol were mixed and homogenized at 3000 r / min for 5 min to obtain phase C;

[0061] (5) Mix laurocapram with the remaining 2 parts of glycerol to obtain phase D;

[0062] (6) Dissolve genipin in 5 parts water and calcium chloride in 5 parts water to obtain genipin solution and calcium chloride solution;

[0063] (7) Add phase B to phase A and stir evenly at 50 r / min. Then add phase C and continue stirring at 50 r / min for 10 min. Then add genipin solution and react for 10 min. Then quickly add calcium chloride solution to obtain a light yellow transparent gel.

[0064] (8) Add phase D to the pale yellow transparent gel and add the remaining 26 parts of water. Stir evenly at 300 r / min, perform vacuum degassing (-0.1 Mpa, 10 min), and dispense to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0065] Example 2

[0066] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0067] (1) Weigh the following raw materials in the following proportions by weight: 3 parts baicalin, 2 parts berberine, 4 parts asiaticoside, 5 parts salvianolic acid B, 3 parts gallic acid, 7 parts bletilla polysaccharide, 5 parts konjac glucomannan, 4 parts chitosan, 12 parts glycerol, 1 part genipin, 2 parts calcium chloride, 2 parts laurocapram, and 50 parts water.

[0068] (2) Mix Bletilla striata polysaccharide, konjac glucomannan and 25 parts of water preheated to 57°C, stir at 250 r / min and heat to 82°C, keep warm and stand for 30 min, cool to 30°C to obtain phase A;

[0069] (3) Chitosan is mixed with 10 parts of acidified water (pH 4.5, the acidification is carried out with 2wt% glacial acetic acid) and stirred at 400 r / min until completely dissolved to obtain phase B;

[0070] (4) Baicalin, berberine, asiaticoside, salvianolic acid B, gallic acid and 9 parts of glycerol were mixed and homogenized at 3500 r / min for 8 min to obtain phase C;

[0071] (5) Mix laurocapram with the remaining 3 parts of glycerol to obtain phase D;

[0072] (6) Dissolve genipin in 5 parts water and calcium chloride in 5 parts water to obtain genipin solution and calcium chloride solution;

[0073] (7) Add phase B to phase A and stir evenly at 80 r / min. Then add phase C and continue stirring at 80 r / min for 12 min. Then add genipin solution and react for 15 min. Then quickly add calcium chloride solution to obtain a light yellow transparent gel.

[0074] (8) Add phase D to the pale yellow transparent gel and add the remaining 5 parts of water. Stir evenly at 400 r / min, perform vacuum degassing (-0.08 Mpa, 13 min), and dispense to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0075] Example 3

[0076] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0077] (1) Weigh the following raw materials in the following proportions by weight: 5 parts baicalin, 3 parts berberine, 6 parts asiaticoside, 6 parts salvianolic acid B, 4 parts gallic acid, 5 parts bletilla polysaccharide, 3 parts konjac glucomannan, 3 parts chitosan, 15 parts glycerol, 1 part genipin, 1 part calcium chloride, 3 parts laurocapram, and 45 parts water.

[0078] (2) Mix Bletilla striata polysaccharide, konjac glucomannan and 20 parts of water preheated to 60°C, stir at 300 r / min and heat to 85°C, keep warm and stand for 35 min, cool to 40°C to obtain phase A;

[0079] (3) Chitosan is mixed with 8 parts of acidified water (pH 5, the acidification is carried out with 3 wt% glacial acetic acid) and stirred at 500 r / min until completely dissolved to obtain phase B;

[0080] (4) Baicalin, berberine, asiaticoside, salvianolic acid B, gallic acid and 10 parts of glycerol were mixed and homogenized at 4000 r / min for 10 min to obtain phase C;

[0081] (5) Mix laurocapram with the remaining 5 parts of glycerol to obtain phase D;

[0082] (6) Dissolve genipin in 5 parts water and calcium chloride in 5 parts water to obtain genipin solution and calcium chloride solution;

[0083] (7) Add phase B to phase A and stir at 100 r / min until homogeneous. Then add phase C and continue stirring at 100 r / min for 15 min. Then add genipin solution and react for 20 min. Then quickly add calcium chloride solution to obtain a pale yellow transparent gel.

[0084] (8) Add phase D to the pale yellow transparent gel and add the remaining 7 parts of water. Stir evenly at 500 r / min, perform vacuum degassing (-0.05 Mpa, 15 min), and dispense to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0085] Comparative Example 1

[0086] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0087] Compared to Example 2, the only difference is the omission of baicalin and gallic acid:

[0088] (1) Weigh the following raw materials in the following proportions by weight: 2 parts berberine, 4 parts asiaticoside, 5 parts salvianolic acid B, 7 parts bletilla polysaccharide, 5 parts konjac glucomannan, 4 parts chitosan, 12 parts glycerol, 1 part genipin, 2 parts calcium chloride, 2 parts laurocapram, and 50 parts water.

[0089] (2) Mix Bletilla striata polysaccharide, konjac glucomannan and 25 parts of water preheated to 57°C, stir at 250 r / min and heat to 82°C, keep warm and stand for 30 min, cool to 30°C to obtain phase A;

[0090] (3) Chitosan is mixed with 10 parts of acidified water (pH 4.5, the acidification is carried out with 2wt% glacial acetic acid) and stirred at 400 r / min until completely dissolved to obtain phase B;

[0091] (4) Mix berberine, asiaticoside, salvianolic acid B with 9 parts of glycerol and homogenize at 3500 r / min for 8 min to obtain phase C;

[0092] (5) Mix laurocapram with the remaining 3 parts of glycerol to obtain phase D;

[0093] (6) Dissolve genipin in 5 parts water and calcium chloride in 5 parts water to obtain genipin solution and calcium chloride solution;

[0094] (7) Add phase B to phase A and stir evenly at 80 r / min. Then add phase C and continue stirring at 80 r / min for 12 min. Then add genipin solution and react for 15 min. Then quickly add calcium chloride solution to obtain a light yellow transparent gel.

[0095] (8) Add phase D to the pale yellow transparent gel and add the remaining 5 parts of water. Stir evenly at 400 r / min, perform vacuum degassing (-0.08 Mpa, 13 min), and dispense to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0096] Comparative Example 2

[0097] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0098] Compared with Example 2, the only difference is that baicalin is replaced with matrine and gallic acid is replaced with paeonol.

[0099] (1) Weigh the following raw materials in the following proportions by weight: matrine 3 parts, berberine 2 parts, asiaticoside 4 parts, salvianolic acid B 5 parts, paeonol 3 parts, bletilla polysaccharide 7 parts, konjac glucomannan 5 parts, chitosan 4 parts, glycerol 12 parts, genipin 1 part, calcium chloride 2 parts, laurocapram 2 parts, and water 50 parts.

[0100] (2) Mix Bletilla striata polysaccharide, konjac glucomannan and 25 parts of water preheated to 57°C, stir at 250 r / min and heat to 82°C, keep warm and stand for 30 min, cool to 30°C to obtain phase A;

[0101] (3) Chitosan is mixed with 10 parts of acidified water (pH 4.5, the acidification is carried out with 2wt% glacial acetic acid) and stirred at 400 r / min until completely dissolved to obtain phase B;

[0102] (4) Mix matrine, berberine, asiaticoside, salvianolic acid B, paeonol with 9 parts of glycerol and homogenize at 3500 r / min for 8 min to obtain phase C;

[0103] (5) Mix laurocapram with the remaining 3 parts of glycerol to obtain phase D;

[0104] (6) Dissolve genipin in 5 parts water and calcium chloride in 5 parts water to obtain genipin solution and calcium chloride solution;

[0105] (7) Add phase B to phase A and stir evenly at 80 r / min. Then add phase C and continue stirring at 80 r / min for 12 min. Then add genipin solution and react for 15 min. Then quickly add calcium chloride solution to obtain a light yellow transparent gel.

[0106] (8) Add phase D to the pale yellow transparent gel and add the remaining 5 parts of water. Stir evenly at 400 r / min, perform vacuum degassing (-0.08 Mpa, 13 min), and dispense to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0107] Comparative Example 3

[0108] A method for preparing a composition for treating intractable skin infections, sores, and inflammation, comprising the following steps:

[0109] Compared to Example 2, the difference lies in the preparation process, which is simply physical mixing:

[0110] (1) Weigh the following raw materials in the following proportions by weight: 3 parts baicalin, 2 parts berberine, 4 parts asiaticoside, 5 parts salvianolic acid B, 3 parts gallic acid, 7 parts bletilla polysaccharide, 5 parts konjac glucomannan, 4 parts chitosan, 12 parts glycerol, 1 part genipin, 2 parts calcium chloride, 2 parts laurocapram, and 50 parts water.

[0111] (2) Mix the above raw materials, stir evenly at 400 r / min, perform vacuum degassing (-0.08 Mpa, 13 min), and package to obtain the composition for treating stubborn skin infections, sores and inflammation.

[0112] Experimental Example 1

[0113] In vitro release rate study:

[0114] The Franz diffusion cell method was used, with 0.9% phosphate-buffered saline (PBS, pH 7.4) as the release medium and a temperature of 37 ± 0.5 °C. The gel products obtained in Example 2 and Comparative Example 3 were placed in the supply cell, respectively. Samples were taken at predetermined time points (1, 2, 4, 6, 8, 12, 24 h), and the concentration of salvianolic acid B was determined by HPLC. The cumulative release rate (%) was calculated. The results are shown in Table 1.

[0115] Table 1. Cumulative release rate of salvianolic acid B (%, n=3)

[0116] Time (h) 1 2 4 6 8 12 24 Example 2 12.3±1.5 21.1±2.1 33.4±1.8 42.7±2.3 52.6±1.9 63.2±1.3 75.8±2.5 Comparative Example 3 45.6±3.2 68.9±2.8 82.1±2.5 87.5±1.9 89.2±1.7 90.5±1.5 91.8±1.2

[0117] As shown in Table 1, in Comparative Example 3, salvianolic acid B was rapidly released at a rate exceeding 85% within 8 hours, followed by a plateau in release. In contrast, the gel product of Example 3 of this application exhibited a stable sustained-release trend within 24 hours, with a cumulative release rate of 75.8%. These results demonstrate that, compared to a simple physical mixing process, the stepwise in-situ crosslinking preparation process employed in this application successfully constructed a sustained-release system, significantly prolonging the drug's duration of action.

[0118] Experimental Example 2

[0119] In vitro antibacterial test:

[0120] The in vitro antibacterial activity of the products in Examples 1-3 and Comparative Examples 1-3 against drug-resistant bacteria was evaluated using the agar diffusion method.

[0121] 1. Experimental bacterial strains:

[0122] Methicillin-resistant Staphylococcus aureus (MRSA, ATCC 43300, purchased from Baosai Plasmid Strains Resources Company);

[0123] Pseudomonas aeruginosa (ATCC 27853, purchased from Qingdao Haibo Biotechnology Co., Ltd.).

[0124] 2. Experimental culture medium:

[0125] Mueller-Hinton (MH) agar medium.

[0126] 3. Experimental group processing:

[0127] Example 1: Dilute with sterile water 10 times;

[0128] Example 2: Diluted 10 times with sterile water;

[0129] Example 3: Dilute with sterile water 10 times;

[0130] Comparative Example 1: Diluted 10 times with sterile water;

[0131] Comparative Example 2: Diluted 10 times with sterile water;

[0132] Comparative Example 3: Diluted 10 times with sterile water;

[0133] Negative control group: sterile saline;

[0134] Positive control group: Mupirocin ointment, diluted 10 times with sterile water.

[0135] 4. Experimental methods:

[0136] The prepared MRSA bacterial suspension (concentration 1×10⁻⁶) was applied using a coating method. 8 The product solution (CFU / mL) was evenly spread onto MH agar plates. 6mm diameter wells were punched in the agar using a sterile puncher, and the agar was removed from the wells using a sterile needle. An equal volume (50μL) of each product solution was accurately added to the center of each well using a micropipette. The plates were placed upright in a 4℃ refrigerator for pre-diffusion for 3 hours, then transferred to a 37℃ incubator for 24 hours. The diameter of each inhibition zone (including the well diameter) was measured using calipers, and the average of two cross-sectional measurements was taken, accurate to 0.1mm. Three replicates were set for each group.

[0137] The experimental results are shown in Table 2.

[0138] Table 2. Results of inhibition zone diameter measurements for different bacterial species in each group (n=3, mm)

[0139] Grouping Methicillin-resistant Staphylococcus aureus Pseudomonas aeruginosa Example 1 17.2±0.5 14.9±0.7 Example 2 18.8±0.7 15.8±0.6 Example 3 17.6±0.9 15.2±0.2 Comparative Example 1 11.8±0.4 9.1±0.3 Comparative Example 2 10.2±0.8 8.5±0.5 Comparative Example 3 12.3±0.6 9.8±0.2 negative control group 6.0±0.0 (hole diameter only) 6.0±0.0 Positive control group 20.5±0.9 6.0±0.0 (hole diameter only)

[0140] Table 2 shows that the negative control group had no inhibition zone, indicating that the experimental system is reliable. Compared with comparative examples 1-3, the inhibition zones of examples 1-3 of this application are significantly larger, with example 2 showing the best effect. Compared with the first-line drug mupirocin ointment, the products of examples 1-3 of this application not only have excellent activity comparable to mupirocin against methicillin-resistant Staphylococcus aureus, but also effectively combat Pseudomonas aeruginosa, which is completely ineffective against mupirocin, exhibiting a broader antibacterial spectrum. This result intuitively demonstrates that this application, through specific component compatibility and preparation process, produces a significant synergistic antibacterial effect among the components.

[0141] Experimental Example 3

[0142] Evaluation of the anti-inflammatory effect of LPS-induced macrophage inflammation model:

[0143] 1. Experimental Method:

[0144] An inflammation model was established by stimulating mouse macrophages (RAW264.7) with lipopolysaccharide (LPS, 1 μg / mL). After intervention with different drugs, the cells were cultured for 24 h. The concentrations of key inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the cell supernatant were detected using ELISA.

[0145] 2. Experimental Groups:

[0146] Example 2: LPS stimulation + Example 2 product;

[0147] Comparative Example 1: LPS stimulation + Comparative Example 1 product;

[0148] Comparative Example 2: LPS stimulation + Comparative Example 2 product;

[0149] Comparative Example 3: LPS stimulation + Comparative Example 3 product;

[0150] Blank control group: Cells were cultured normally, without LPS stimulation or any drugs;

[0151] Model control group: Cells were stimulated with LPS, but no treatment drugs were added.

[0152] The experimental results are shown in Tables 3 and 4.

[0153] Table 3 shows the effect of LPS on the release of inflammatory factors in RAW264.7 cells.

[0154] Grouping TNF-α concentration (pg / mL) IL-6 concentration (pg / mL) Blank control group 85.5 65.3 Model control group 1250.8 980.5 Example 2 275.4 270.8 Comparative Example 1 740.2 625.8 Comparative Example 2 810.5 670.2 Comparative Example 3 562.3 505.4

[0155] Based on the above data, the inhibition rate (%) of each group relative to the model control group was calculated:

[0156] Inhibition rate (%) = [1 - (concentration of inflammatory factors measured in each treatment group - concentration of inflammatory factors measured in the blank control group) / (concentration of inflammatory factors measured in the model control group - concentration of inflammatory factors measured in the blank control group)] × 100%.

[0157] Table 4. Inhibition rate (%) of different products on the release of inflammatory factors

[0158] Grouping TNF-α inhibition rate IL-6 inhibition rate Example 2 83.7 77.5 Comparative Example 1 43.8 38.8 Comparative Example 2 37.8 33.9 Comparative Example 3 59.1 51.9

[0159] As shown in Table 3, compared with the blank control group, the concentrations of TNF-α and IL-6 in the model control group were significantly increased, indicating that the LPS inflammation model was successfully established. Combined with Table 4, it can be seen that compared with comparative examples 1-3, the product of Example 2 of this application had the lowest concentration of inflammatory factors and a significantly higher inhibition rate. This indicates that, through specific component compatibility and preparation processes, this application achieves a significant synergistic anti-inflammatory effect among the active ingredients, rather than simply an additive effect of mixing. This application can effectively inhibit the release of key inflammatory factors from macrophages, providing a solid cellular basis for its ability to alleviate excessive inflammatory responses in wounds in vivo.

[0160] Experiment Example 4

[0161] Cell scratch assay to evaluate the effect on promoting healing:

[0162] 1. Experimental Principle

[0163] Simulated wound: Create a cell-free "scratch zone" on a culture plate covered with cells to simulate a skin wound.

[0164] Intervention: Add culture medium containing different drugs.

[0165] Observation of repair: Cells gradually migrate and proliferate towards the scratched area until the scratch closes. By measuring the changes in scratch width at different time points, the "healing rate" can be quantitatively calculated, thereby determining the drug's ability to promote repair.

[0166] 2. Experimental Materials and Reagents

[0167] Cell line: Mouse fibroblasts (L929).

[0168] Culture medium: DMEM high glucose medium, supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics.

[0169] Experimental equipment: 6-well plate, 200μL sterile pipette tip, microscope equipped with digital camera.

[0170] 3. Experimental Grouping

[0171] Negative control group: Complete culture medium containing 0.1% DMSO (solvent control).

[0172] Positive control group: Complete culture medium containing 10 ng / mL epidermal growth factor (EGF).

[0173] Example 2 and Comparative Examples 1-3: Each of Example 2 and Comparative Examples was extracted with complete culture medium (100 mg / mL), centrifuged, and the supernatant was filtered and sterilized before use (i.e., drug-containing culture medium).

[0174] 4. Experimental Procedure

[0175] Cell seeding: Seed cells in 6-well plates and culture until 100% confluence.

[0176] Create a scratch: Using a 200μL sterile pipette tip, hold the pipette perpendicular to the bottom of the well plate and smoothly create a straight scratch. Ensure all scratches are of uniform width.

[0177] Washing: Gently wash 3 times with PBS to remove floating cell debris.

[0178] Drug treatment: Add drug-containing culture media containing negative control, positive control, Example 2 and Comparative Examples 1-3 respectively.

[0179] Observation and photography: The culture plate was placed in a 37℃, 5% CO2 incubator. At 0h, 12h, and 24h, the scratches were photographed from the same position under a microscope to record the condition.

[0180] Image analysis: Use image analysis software such as ImageJ to measure the width of the scratch at each time point (multiple points can be measured and the average value is taken).

[0181] 5. Data Processing and Result Analysis

[0182] Calculate the scratch healing rate:

[0183] Healing rate (%) = [(W0-W t ) / W0]×100%

[0184] W0: Average width of scratch at 0 hours

[0185] W t Average scratch width at a specific time point (12h, 24h)

[0186] Statistical analysis: All experiments were repeated 3 times.

[0187] The results are shown in Table 5.

[0188] Table 5. The promoting effect of different treatments on fibroblast scratch healing rate

[0189] Grouping 12-hour healing rate (%) 24-hour healing rate (%) negative control group 25.3±3.5 55.1±4.2 Positive control group 45.8±4.1 85.6±3.8 Example 2 65.5±5.2 95.8±2.5 Comparative Example 1 42.1±3.4 72.3±1.8 Comparative Example 2 38.7±5.7 67.9±4.5 Comparative Example 3 46.3±1.5 78.2±2.6

[0190] Table 5 shows that the negative control group cells had some migration ability, with a healing rate of approximately 55.1% after 24 hours. The positive control group (EGF) showed a significant healing-promoting effect, with a healing rate of 85.6% after 24 hours, proving the effectiveness of the experimental system. Compared with comparative examples 1-3, the composition group of Example 2 of this application showed the strongest healing-promoting ability, with significantly higher healing rates (65.5% and 95.8%) at 12 and 24 hours compared to other groups. This indicates that the composition of this application can significantly promote the migration and proliferation of fibroblasts, thereby accelerating wound closure. This provides cellular evidence for its in vivo efficacy in promoting granulation tissue formation and epithelial repair.

[0191] The above description is merely a few embodiments of this application and is not intended to limit this application in any way. Although this application discloses preferred embodiments as described above, it is not intended to limit this application. Any changes or modifications made by those skilled in the art without departing from the scope of the technical solution of this application using the disclosed technical content are equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A composition for treating intractable skin infections, sores, and inflammation, characterized in that, It is prepared from the following components in parts by weight: 1-5 parts baicalin, 0.5-3 parts antibacterial alkaloids, 2-6 parts asiaticoside, 3-8 parts salvianolic acid B, 1-4 parts gallic acid, 5-10 parts Bletilla striata polysaccharide, 3-10 parts konjac glucomannan, 3-5 parts chitosan, 8-20 parts glycerol, 0.5-2 parts genipin, 1-3 parts calcium chloride, 1-3 parts laurocapram, and 40-80 parts water; The antibacterial alkaloids include berberine or phellodendron chinense.

2. The composition for treating intractable skin infections, sores, and inflammation according to claim 1, characterized in that, The degree of deacetylation of the chitosan is ≥85%, and the molecular weight of the chitosan is 5000-20000 Da.

3. A method for preparing a composition for treating intractable skin infections, sores, and inflammation as described in claim 1 or 2, characterized in that, Includes the following steps: (1) Mix Bletilla striata polysaccharide, konjac glucomannan and water preheated to 55-60°C, stir and heat to 80-85°C, keep warm and stand, cool to obtain phase A; (2) Mix chitosan with acidified water and stir until completely dissolved to obtain phase B; (3) Baicalin, antibacterial alkaloids, berberine, asiaticoside, salvianolic acid B, gallic acid and glycerol were mixed and homogenized to obtain phase C; (4) Mix laurocapram with glycerol to obtain phase D; (5) Dissolve genipin and calcium chloride in water to obtain genipin solution and calcium chloride solution respectively; (6) Add phase B to phase A and stir evenly. Then add phase C and continue stirring for 10-15 minutes. Then add genipin solution and react for 10-20 minutes. Finally, add calcium chloride solution to obtain a light yellow transparent gel. (7) Add phase D to the pale yellow transparent gel, add the remaining water, stir evenly, and perform vacuum degassing to obtain the composition for treating stubborn skin infections, sores and inflammation.

4. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, In step (1), the stirring speed is 200-300 r / min; The heat preservation and static setting time is 25-35 minutes; The cooling process involves cooling the temperature to 25–40°C.

5. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, In step (2), the pH of the acidified water is 4 to 5; The acidification was carried out using glacial acetic acid; The concentration of the glacial acetic acid is 1-3 wt%. The stirring speed is 300-500 r / min.

6. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, In step (3), the antibacterial alkaloids include berberine or phellodendron chinense; The rotational speed of the homogenizer is 3000–4000 r / min; The homogenization time is 5 to 10 minutes.

7. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, The ratio of the amount of glycerol used in step (3) to the amount of glycerol used in step (4) is 1 to 3:

1.

8. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, In step (6), the stirring speed is 50-100 r / min.

9. A method for preparing a composition for treating intractable skin infections, sores, and inflammation according to claim 3, characterized in that, In step (7), the stirring speed is 300-500 r / min; The vacuum degassing pressure is -0.1 to -0.05 MPa; The vacuum degassing time is 10-15 minutes.

10. The use of the composition according to claim 1 or 2, or the composition prepared by the preparation method according to any one of claims 3 to 9, in the preparation of a medicament for treating intractable skin infections, sores and inflammation.