Perianal nursing composition containing antibacterial peptide and preparation method of perianal nursing composition

Abstract

The invention provides a crissum nursing composition containing antibacterial peptide and a preparation method, and belongs to the technical field of composition medical nursing. The perianal care composition is prepared from the following raw materials: silk fibroin hydrolysate, carbomer, hyaluronic acid, glycerol, phenoxyethanol, cecropin, melittin, scorpion peptide ABP-W1 and deionized water. According to the crissum care composition, through the synergistic effect of the cecropin, the melittin, the silk fibroin, the carbomer and the hyaluronic acid, a protective film with antibacterial and bacteriostatic, physical barrier, moisturizing and repairing and anti-inflammatory functions is formed in the crissum, and crissum skin mucosa injury and infection caused by diarrhea, radiotherapy and chemotherapy, diaper stimulation and the like can be effectively prevented and treated; the traditional Chinese medicine composition has good curative effect and safety, and is suitable for infants, old people, radiotherapy and chemotherapy sensitive people and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of medical care, and relates to an anal perianal care composition containing antimicrobial peptides and a preparation method thereof. Background Art

[0002] In clinical medical and daily care scenarios, the fragility and vulnerability of the skin and mucous membranes around the anus make this area susceptible to damage from a variety of factors. For example, during diarrhea, frequent bowel movements and the increased digestive enzymes in the feces are highly irritating to the skin, which can damage the skin's barrier function and cause redness, swelling, pain, and even erosion. During radiotherapy, ionizing radiation can cause DNA damage to tissue cells, leading to cell necrosis and tissue fibrosis, which changes the structure and function of the skin and mucous membranes around the anus, making them fragile and vulnerable. When the immune system is weakened, the body's immune defenses are weakened and cannot effectively resist the invasion of pathogens, making the area around the anus more susceptible to infection by bacteria, fungi, etc., causing inflammation and ulcers.

[0003] Currently available care products mainly include oily ointments and aqueous products. Although oily ointments have certain moisturizing and lubricating effects, they have poor air permeability. Long-term use will result in the inability of carbon dioxide on the skin surface to be discharged and oxygen to enter, thereby causing hypoxia and metabolic disorders in skin cells, further aggravating skin damage. In addition, oily ointments have poor adhesion and are easily washed away by feces, and cannot form a long-lasting and effective protective layer on the damaged area. Although aqueous products can quickly penetrate into the skin surface, they have a short residence time on the skin surface when faced with frequent bowel movements and fecal flushing, making it difficult to exert long-term repair and antibacterial effects. Therefore, existing care products mainly focus on relieving symptoms and providing temporary protection, and have weak regeneration and repair capabilities for damaged tissues, resulting in limited repair effects. At the same time, existing care products have insufficient antibacterial capabilities and cannot effectively kill pathogens such as drug-resistant bacteria, making it difficult to prevent and control the occurrence of infections.

[0004] In view of the shortcomings of existing perianal care products, there is a need to develop a new care composition with long-lasting moisturizing, mucosal repair, physical barrier and antibacterial functions, which has important clinical significance and market demand. Summary of the Invention

[0005] The purpose of the present invention is to provide an anal perianal care composition containing antimicrobial peptides and a preparation method thereof, so as to solve the problems that existing care products have poor care effects and are prone to infection under complex pathological environments.

[0006] To achieve the above object, the present invention adopts the following technical solutions: The present application provides an anal perianal care composition containing antimicrobial peptides. The raw materials for preparing the composition include, in percentage by mass: 1-5% silk fibroin hydrolyzate, 0.5-2% carbomer, 0.1-1% hyaluronic acid, 5-10% glycerol, 0.1-0.5% phenoxyethanol, 0.05-0.2% cecropin, 0.03-0.15% melittin, 0.01-0.2% scorpion peptide ABP-W1, and the remainder is deionized water.

[0007] Silk fibroin hydrolysate is a small peptide extracted from mulberry silk using the subtilisin enzymatic hydrolysis method, with a molecular weight of 5,000-15,000 Da. Silk fibroin hydrolysate can penetrate the stratum corneum and reorganize into a β-pleated structure, forming a breathable physical barrier that effectively isolates external irritants such as feces and protects the perianal skin and mucosa. It can also activate the TLR2 pathway in fibroblasts, promote collagen synthesis, and accelerate the repair of damaged tissue. Furthermore, silk fibroin hydrolysate exhibits excellent biocompatibility and moisturizing properties, promoting cell adhesion and proliferation, and providing an excellent material foundation for the repair of the perianal skin and mucosa.

[0008] Carbomer, as a hydrogel matrix, can form a network structure to load active ingredients, allowing them to be stably released at the site of injury and prolong the duration of action.

[0009] Hyaluronic acid, also known as hyaluronic acid, plays the role of water retention, maintaining extracellular space, regulating osmotic pressure, lubrication, and promoting cell repair in the body. In addition, hyaluronic acid can also lock in moisture and regulate inflammatory responses, help maintain the hydration state of the skin and mucous membranes, and promote the regeneration of damaged tissues. In this application, the molecular weight of hyaluronic acid is 400,000. The hyaluronic acid of this molecular segment has high permeability, can promote tyrosine phosphorylation, activate phospholipase PLCγ1, thereby activating the protease signal transduction pathway and the mitogen-activated protein kinase signal transduction system, repair damaged cells, and enhance cell vitality.

[0010] In the present application, the hyaluronic acid is a compound of low molecular weight hyaluronic acid with a molecular weight of 100-500 kDa and high molecular weight hyaluronic acid with a molecular weight of 1000-2000 kDa, and the mass ratio of the low molecular weight hyaluronic acid to the high molecular weight hyaluronic acid is 1:1-3:1.

[0011] Cecropin is a protein with antimicrobial activity that rapidly kills bacteria, particularly Gram-negative bacteria like Escherichia coli, with a kill rate of up to 99.2% within 30 seconds. Melittin, a substance with multiple biological activities, including antimicrobial and anti-inflammatory properties, can destroy fungal cell walls and exhibit significant inhibitory effects against fungi such as Candida albicans, with an inhibition zone diameter exceeding 15 mm. Melittin and cecropin work synergistically to rapidly kill a variety of drug-resistant pathogens, including Gram-negative and Gram-positive bacteria, effectively preventing and controlling perianal infections.

[0012] Scorpion peptide ABP-W1 is a cationic tridecapeptide with a molecular weight of 1357.8 Da and an amino acid sequence of Gly-Ile-Gly-Ala-Phe-Leu-Lys-Lys-Leu-Phe-Lys-Lys-Ile. Scorpion peptide ABP-W1 can target and disrupt the phospholipid bilayer of microbial membranes, with a minimum inhibitory concentration (MIC) as low as 2 μg / mL against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus (MSSA). Metabolites of scorpion peptide ABP-W1 are naturally occurring amino acids, which have been shown to be non-sensitizing in rabbit skin irritation tests and meet ISO 10993-10 standards.

[0013] The perianal care composition provided by the present application forms a triple protection mechanism through the synergistic effect of the above-mentioned components. The first layer utilizes the targeted bactericidal effect of antimicrobial peptides such as scorpion peptide ABP-W1, cecropin and melittin to quickly kill various drug-resistant pathogens such as Gram-negative bacteria and Gram-positive bacteria, effectively preventing and controlling perianal infections. The second layer, silk fibroin hydrolyzate and hyaluronic acid cooperate with each other to form a breathable physical barrier, isolate external stimuli, and promote the repair and regeneration of damaged cells. The third layer, the film-forming property and physical isolation effect of carbomer further enhance the protective effect of the composition on the perianal skin and mucosa. Thus, the composition can effectively prevent and treat perianal skin and mucosal damage and infection caused by diarrhea, radiotherapy and chemotherapy, diaper irritation, etc., with good efficacy and safety.

[0014] Preferably, the raw materials for preparing the perianal care composition include, in percentage by mass: 3.2% silk fibroin hydrolyzate, 1.2% carbomer, 0.5% hyaluronic acid, 8% glycerol, 0.3% phenoxyethanol, 0.1% cecropin, 0.08% bee venom, 0.05% scorpion peptide ABP-W1 and 86.57% deionized water, wherein the hyaluronic acid is a compound of low molecular weight hyaluronic acid and high molecular weight hyaluronic acid in a mass ratio of 2:1.

[0015] In the present application, the dosage form of the perianal care composition includes a hydrogel, an ointment, a spray or a film, which is used to prevent or treat perianal skin and mucosal damage and infection caused by diarrhea, chemotherapy, and diaper irritation, wherein the symptoms of damage and infection include perianal redness, fissures, ulcers, Candida infection or MRSA infection.

[0016] The present application also provides a method for preparing an anal perianal care composition containing antimicrobial peptides, comprising: S01: Add glycerin and hyaluronic acid to the swollen carbomer and stir evenly to form a matrix system.

[0017] Disperse the carbomer powder in 30%-50% deionized water and stir at 800-1200 rpm for 40-60 minutes until fully swollen. This ensures the carbomer forms a uniform gel network structure within the composition, providing good rheological properties for subsequent processing and use. Glycerin and hyaluronic acid are then added to the swollen carbomer, followed by stirring at 300-500 rpm for 20 minutes at a constant temperature of 45±2°C until uniform, allowing all components to fully mix and form a stable matrix system.

[0018] S02: adding silk fibroin hydrolyzate, phenoxyethanol, cecropin, melittin and scorpion peptide ABP-W1 to the matrix system, adjusting the pH value to 5.5-6.5, and obtaining anal perianal care composition.

[0019] After the matrix system is cooled to 25-30°C, silk fibroin protein hydrolyzate, phenoxyethanol, cecropin, melittin and scorpion peptide ABP-W1 are added, and triethanolamine is used to adjust the pH value to 5.5-6.5 to ensure the stability and biological activity of the active ingredients in the composition. At the same time, the pH value of the composition is also made close to the physiological pH value of human skin and mucous membranes, thereby improving the comfort and safety of use, thereby obtaining an anal care composition.

[0020] The present invention has the following beneficial effects: (1) Antibacterial and antimicrobial effects: Cecropin, bee toxin and scorpion peptide ABP-W1 have significant inhibitory effects on common perianal pathogens such as Escherichia coli, Staphylococcus aureus and Candida albicans by destroying the bacterial cell membrane structure and interfering with intracellular metabolism. The antibacterial rate can reach more than 95%.

[0021] (2) Isolation and protection: Silk fibroin forms a nanoscale protective film to isolate the chemical stimulation of digestive enzymes and acidic substances in feces.

[0022] (3) Moisturizing and repairing: By constructing a three-dimensional hydration network through hyaluronic acid, the moisturizing rate of the perianal skin is increased by more than 40%, promoting the repair of damaged mucosal epithelium.

[0023] (4) Reduce friction: Carbomer gel matrix has adhesive properties and can form long-term adhesion in the perianal folds, with a retention time of ≥8 hours, thereby reducing mechanical friction damage.

[0024] (5) Biosafety: The ingredients used in the preparation of the perianal care composition are all natural extracts or pharmaceutical excipients, without the addition of alcohol, hormones, etc. It is suitable for sensitive groups such as infants, the elderly, and those undergoing radiotherapy and chemotherapy.

[0025] (6) Applicability in multiple scenarios: It has significant effects on infant diaper rash, elderly incontinence dermatitis, perianal care for cancer patients, perianal redness and swelling caused by diarrhea, mucosal ulcers after radiotherapy, and diaper dermatitis. It can also effectively prevent and treat local infections. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a graph comparing the cytotoxicity of the perianal care composition prepared in Example 1 of the present application on mucosal epithelial cells; Figure 2 This is a diagram showing the effect of the perianal care composition prepared in Example 1 of the present application on repairing perianal injuries in rats; Figure 3 This is a graph showing the effect of the perianal care composition prepared in Example 1 of the present application on the inflammatory factor TNF-α in rats; Figure 4 This is a graph showing the effect of the perianal care composition prepared in Example 1 of the present application on the inflammatory factor IL-10 in rats; Figure 5 This is a graph showing the effect of the perianal care composition prepared in Example 1 of the present application on the antioxidant SOD in rats; Figure 6 This is a graph showing the effect of the perianal care composition prepared in Example 1 of the present application on the antioxidant MDA in rats. DETAILED DESCRIPTION

[0027] The technical solution of the present invention is further explained and illustrated by means of specific embodiments below.

[0028] Example 1 An embodiment of the present application provides an anal perianal care composition containing antimicrobial peptides, wherein the raw materials for preparing the composition include, in percentage by mass: 3.2% silk fibroin hydrolyzate, 1.2% carbomer, 0.5% hyaluronic acid, 8% glycerol, 0.3% phenoxyethanol, 0.1% cecropin, 0.08% bee venom, 0.05% scorpion peptide ABP-W1, and 86.57% deionized water, wherein the hyaluronic acid is a compound formed by low-molecular-weight hyaluronic acid and high-molecular-weight hyaluronic acid in a mass ratio of 2:1, and the molecular weights of the low-molecular-weight hyaluronic acid and the high-molecular-weight hyaluronic acid are 400 kDa and 1600 kDa, respectively.

[0029] The present application also provides a method for preparing an anal perianal care composition containing antimicrobial peptides, the method comprising: S101: Disperse carbomer powder in 30% deionized water and stir at 1000 rpm for 60 minutes until fully swollen. Add glycerin and hyaluronic acid to the swollen carbomer in sequence and stir at 400 rpm for 20 minutes at a constant temperature of 45±2°C until uniform, ensuring thorough mixing of the components and forming a stable matrix system.

[0030] S102: Cool the matrix system to 25° C., then add silk fibroin hydrolyzate, phenoxyethanol, cecropin, melittin and scorpion peptide ABP-W1, and adjust the pH value to 6.0±0.2 with triethanolamine to form an anal care composition.

[0031] The perianal care composition provided in the embodiments of the present application is a universal care composition with a viscosity of 3000-5000 mPa•s. It is suitable for protection and care in scenarios such as daily diarrhea and mild perianal discomfort. The inhibition rate of Escherichia coli can reach 96%. After 3 days of use, the symptom relief rate of patients with mild redness and swelling reaches 88%.

[0032] Example 2 An embodiment of the present application provides an anal perianal care composition containing antimicrobial peptides, wherein the raw materials for preparing the composition include, in percentage by mass: 5% silk fibroin hydrolyzate, 2% carbomer, 1% hyaluronic acid, 10% glycerol, 0.5% phenoxyethanol, 0.2% cecropin, 0.15% bee venom, 0.2% scorpion peptide ABP-W1, and 80.95% deionized water, wherein the hyaluronic acid is a compound formed by low-molecular hyaluronic acid and high-molecular hyaluronic acid in a mass ratio of 3:1, and the molecular weights of the low-molecular hyaluronic acid and the high-molecular hyaluronic acid are 500 kDa and 2000 kDa, respectively.

[0033] The preparation method of the anal perianal care composition containing antimicrobial peptides provided in the examples of this application is the same as that in Example 1.

[0034] The perianal care composition provided in the embodiments of the present application is a deep-repair antibacterial composition with a viscosity of 8000-10000 mPa•s. It is suitable for scenarios such as perianal ulcers after radiotherapy and chemotherapy, and mucosal damage caused by severe diarrhea. The inhibition rate of Staphylococcus aureus can reach 97%. Clinical experiments show that after using this composition, the healing time of ulcer patients is shortened by an average of 38%.

[0035] Example 3 An embodiment of the present application provides an anal perianal care composition containing antimicrobial peptides, wherein the raw materials for preparing the composition include, in percentage by mass: 1.5% silk fibroin hydrolyzate, 0.8% carbomer, 0.3% hyaluronic acid, 6% glycerol, 0.2% phenoxyethanol, 0.05% cecropin, 0.03% bee venom, 0.01% scorpion peptide ABP-W1, and 91.11% deionized water, wherein the hyaluronic acid is a compound formed by low-molecular hyaluronic acid and high-molecular hyaluronic acid in a mass ratio of 1:1, and the molecular weights of the low-molecular hyaluronic acid and the high-molecular hyaluronic acid are 100 kDa and 1000 kDa, respectively.

[0036] The preparation method of the perianal care composition containing antimicrobial peptides provided in the embodiment of the present application is the same as that in Example 1, except that the pH value is 5.0-5.5 to meet the weakly acidic environment requirement of infant skin.

[0037] The perianal care composition provided in the embodiments of the present application is an antibacterial composition specially designed for infants and young children. Its inhibition rate against Candida albicans can reach 95%, and it is significantly effective in preventing and treating diaper dermatitis in infants and young children, with a redness and swelling relief rate of 90% within 24 hours.

[0038] Example 4 An embodiment of the present application provides an anal perianal care composition containing antimicrobial peptides, wherein the raw materials for preparing the composition include, in percentage by mass: 2% silk fibroin hydrolyzate, 1% carbomer, 0.4% hyaluronic acid, 7% glycerol, 0.25% phenoxyethanol, 0.08% cecropin, 0.05% bee venom, 0.12% scorpion peptide ABP-W1, and 89.1% deionized water, wherein the hyaluronic acid is a compound formed by low-molecular hyaluronic acid and high-molecular hyaluronic acid in a mass ratio of 2:1, and the molecular weights of the low-molecular hyaluronic acid and the high-molecular hyaluronic acid are 200 kDa and 1200 kDa, respectively.

[0039] The preparation method of the anal perianal care composition containing antimicrobial peptides provided in the examples of this application is the same as that in Example 1.

[0040] The perianal care composition provided in the embodiments of the present application is an antibacterial composition specially designed for sensitive skin. It is mild and non-irritating, with a comprehensive inhibition rate of 94% against common pathogens. It can effectively prevent and relieve skin discomfort around the anus caused by friction, irritation, etc.

[0041] In the present embodiment, the anal perianal care composition containing antimicrobial peptides prepared in Example 1 was used to perform mucosal epithelial cell toxicity testing, anal perianal injury repair effect testing, inflammatory factor impact testing, and antioxidant index level testing, which are described in detail below.

[0042] (1) Toxicity test of perianal care composition on mucosal epithelial cells Mucosal epithelial cells cultured to P3-P5 were added to the culture medium and suspended to prepare a cell suspension. 4 Cells were seeded at a density of 100 μL / well in a 96-well plate, with 100 μL of cell suspension in each well. The inoculated plates were placed in an incubator and pre-incubated for 4 hours at 37°C and 5% CO2. The pre-incubated cell suspension was divided into three groups: A, B, and C, with three replicates in each group.

[0043] After the pre-culture, the culture medium was discarded. Group A was a blank group, and a culture medium without a care substance was added; Group B was a control group, and an equal volume of the perianal care composition and PBS in Example 1 was added; Group C was an experimental group, and the perianal care composition in Example 1 was added. Groups ABC were placed in an incubator and cultured at 37°C and 5% CO2 for 48 hours. After the culture was completed, 10ul of CCK-8 reagent was added to each well and the culture was continued for 2 hours. The absorbance value at a wavelength of 450nm was detected using an enzyme marker to determine the cell proliferation rate, and the attached Figure 1 .

[0044] By the attached Figure 1 It can be seen that during the culture process of mucosal epithelial cells, the absorbance value at OD450 of Group A was about 0.720, the absorbance value at OD450 of Group B was about 0.710, and the absorbance value at OD450 of Group C was about 0.715. There was no statistical difference among the three groups. This shows that the perianal care composition prepared in the examples of the present application has no cytotoxicity to mucosal epithelial cells and can maintain good cell activity. This shows that the perianal care composition prepared in the examples of the present application can effectively promote the proliferation of mucosal epithelial cells and play a protective and repairing role on the cells.

[0045] (2) Detection of the effect of the perianal care composition on the repair of perianal injuries in rats Eighteen healthy male Sprague-Dawley rats, aged 8-10 weeks and weighing 200-250 g, were housed in a standard laboratory environment with a temperature of 22 ± 2°C and a humidity of 50 ± 10% under a 12-hour light / dark cycle. They had free access to food and water. The 18 Sprague-Dawley rats were randomly divided into three groups of six rats each: Group A served as the blank group, Group B served as the control group, and Group C served as the experimental group.

[0046] The rats were fasted for 12 hours before the experiment, and fixed in a prone position after isoflurane inhalation anesthesia. The hair in the 2cm×2cm area around the anus was removed with an electric shaver, and the skin was cleaned with a sterile cotton swab dipped in normal saline. Group A was smeared with normal saline, and Groups B and C were soaked with filter paper in 40% acetic acid, and the filter paper was applied to the rat anus for 90 seconds. After removing the filter paper, 20μL of MRSA and Candida albicans mixed bacterial solution was taken with a pipette and added to the damaged area. It was gently spread evenly with a sterile glass rod and allowed to stand for 5 minutes to absorb, completing the perianal injury model. After 24 hours of modeling, Group B was not treated, and Group C was smeared with the perianal care composition prepared in Example 1, twice a day for 7 consecutive days. Pictures were taken after 7 days, and the attached Figure 2 In addition, after the photography was completed, the rats were killed and the inflammatory factors and antioxidant indexes were detected.

[0047] By the attached Figure 2 It can be seen that the rats in Group B had perianal redness, swelling and ulcers, while the rats in Group C had perianal redness and swelling reduced and the anus returned to normal size after treatment with the perianal care composition. This shows that the perianal care composition prepared in Example 1 of the present application can effectively alleviate the inflammatory response of perianal injury, repair damaged tissue, and provide long-term protection to the perianal area.

[0048] (3) Detection of the effect of perianal care composition on inflammatory factors TNF-α and IL-10 The perianal mucosal tissue of the sacrificed rats was collected and 1 mL of pre-cooled PBS homogenization buffer containing protease inhibitors was added. The homogenate was prepared using an ultrasonic disruptor. The homogenate was centrifuged at 12000 rpm for 15 minutes at 4°C and the supernatant was collected. The supernatant was used for TNF-α and IL-10 detection. Figure 3 、 4 The detection process of ELISA-TNF-α and ELISA-IL-10 is the same. The detection process of ELISA-TNF-α is described in detail below: According to the kit instructions, dilute the standard to a concentration series, such as 0, 15.6, 31.2, 62.5, 125, 250, 500, and 1000 pg / mL. Prepare wash buffer, chromogenic substrate, and stop solution. Add 100 µL of each standard and sample to each well of a 96-well plate pre-coated with the antibody. Set up three replicates for each sample and standard. Add only the diluent to the blank control well. After addition, cover the plate with sealing film and incubate at 37°C for 1.5 hours. After incubation, discard the liquid from the wells and add 300 µL of wash buffer to each well. Let it sit for 30 seconds before discarding. Repeat 3-5 times. Discard the wash buffer and add 100 µL of enzyme-linked secondary antibody (HRP-conjugated) to each well. Incubate at 37°C for 1 hour. After incubation, discard the liquid from the wells and wash 3-5 times. Discard the wash buffer and add 100 µL of TMB chromogenic substrate to each well. Incubate at room temperature in the dark for 15-30 minutes. After incubation, add 50 µL of 2 M H₂SO₄ to each well and mix gently to terminate the reaction. Read the absorbance of each well at 450 nm using a microplate reader.

[0049] By the attached Figure 3 、 4 It can be seen that during the inflammatory factor detection process, the concentration of the inflammatory factor TNF-α in group A was about 21pg / mL, and the concentration of the inflammatory factor IL-10 was about 70pg / mL; the concentration of the inflammatory factor TNF-α in group B was about 60pg / mL, and the concentration of the inflammatory factor IL-10 was about 25pg / mL; the concentration of the inflammatory factor TNF-α in group C was about 20pg / mL, and the concentration of the inflammatory factor IL-10 was about 68pg / mL, which was close to that of the normal group A.

[0050] Thus, the perianal care composition provided in Example 1 of the present application can effectively improve the elevated levels of the pro-inflammatory factor TNF-α caused by perianal injury in rats, while simultaneously reducing the level of the anti-inflammatory factor IL-10. This indicates that the perianal care composition in the present application example can effectively reduce inflammation, repair damaged mucosa, and provide long-term protective effects on tissues.

[0051] (4) Detection of SOD and MDA levels of perianal care compositions for antioxidant indicators The perianal mucosal tissue of the sacrificed rats was taken and 1 mL of pre-cooled PBS homogenization buffer containing protease inhibitors was added and homogenized using an ultrasonic disruptor. The homogenate was centrifuged at 12000 rpm for 15 minutes at 4°C and the supernatant was collected. The supernatant was used to detect the SOD activity level using the WST-1 method and the MDA level using the TBA method to obtain the attached Figure 5 、 6 .

[0052] Among them, the process of using the WST-1 method to detect the SOD activity level is as follows: prepare the working solution according to the reagent instructions, add 20μL double-distilled water and 20μL enzyme working solution to the control group, add 20μL double-distilled water and 20μL reagent four to the control blank group, add 20μL of the test sample and 20μL enzyme working solution to the measurement well, use a multi-channel pipette to add 200μL substrate application solution to each well, and carefully blow and mix, incubate at 37℃ for 20min, and detect the OD value at 450nm with a microplate reader.

[0053] The process of using the TBA method to detect MDA levels is as follows: prepare the standard and working solution according to the reagent instructions, take out 0.02 mL of 8 standards of different concentrations, add them to the numbered 1.5 mL EP respectively, add 0.02 mL of the sample to be tested to the sample tube, and add 0.02 mL of the sample to be tested to the control tube; add 0.02 mL of reagent one to each tube, and then add 0.6 mL of reagent two application solution to each tube again; add 0.2 mL of reagent three application solution to the standard tube and the measurement tube, and add 0.2 mL of 50% acetic acid to the control tube; tie all tubes tightly with plastic wrap, mix thoroughly, and place in a 100°C water bath for 40 minutes; after the running water is cooled to room temperature, centrifuge at 9569g for 10 minutes, take 0.25 mL of the supernatant and add it to the ELISA plate, and detect its OD value at 532 nm.

[0054] By the attached Figure 5 、 6 It can be seen that during the antioxidant index detection process, the SOD concentration of group A was about 173 U / mL, and the MDA concentration was about 10 μmol / L; the SOD concentration of group B was about 146 U / mL, and the MDA concentration was about 16 μmol / L; the SOD concentration of group C was about 177 U / mL, and the MDA concentration was about 11 μmol / L.

[0055] It can be seen that the perianal care composition provided in Example 1 of the present application can restore the free radical scavenging ability of damaged tissues to a physiological level, which is slightly better than the normal state. At the same time, it can effectively inhibit the lipid peroxidation chain reaction, with a repair efficiency of 90%.

[0056] In summary, the anal care composition provided in the embodiments of the present application adopts the synergistic effect of cecropin, melittin, silk fibroin, carbomer and hyaluronic acid to form a protective film around the anus that has antibacterial and antibacterial functions, a physical barrier, moisturizing and repairing, and anti-inflammatory functions, effectively preventing and treating anal skin and mucous membrane damage and infection caused by diarrhea, chemotherapy, diaper irritation, etc., and has good efficacy and safety.

[0057] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. An anal perianal care composition containing antimicrobial peptides, characterized in that: The raw materials for the preparation include, by mass percentage, 1-5% of silk fibroin hydrolyzate, 0.5-2% of carbomer, 0.1-1% of hyaluronic acid, 5-10% of glycerol, 0.1-0.5% of phenoxyethanol, 0.05-0.2% of cecropin, 0.03-0.15% of melittin, 0.01-0.2% of scorpion peptide ABP-W1, and the rest is deionized water.

2. The anal perianal care composition containing antimicrobial peptides according to claim 1, characterized in that: The raw materials for the preparation include, by mass percentage, 3.2% of silk fibroin hydrolyzate, 1.2% of carbomer, 0.5% of hyaluronic acid, 8% of glycerol, 0.3% of phenoxyethanol, 0.1% of cecropin, 0.08% of melittin, 0.05% of scorpion peptide ABP-W1, and 86.57% of deionized water.

3. The anal perianal care composition containing antimicrobial peptides according to claim 1, characterized in that: The molecular weight of the silk fibroin hydrolyzate is 5000-15000Da.

4. The anal perianal care composition containing antimicrobial peptides according to claim 1, characterized in that: The hyaluronic acid is formed by compounding low-molecular hyaluronic acid with a molecular weight of 100-500 kDa and high-molecular hyaluronic acid with a molecular weight of 1000-2000 kDa.

5. The anal perianal care composition containing antimicrobial peptides according to claim 4, characterized in that: The mass ratio of the low-molecular hyaluronic acid to the high-molecular hyaluronic acid is 1:1-3:

1.

6. The anal perianal care composition containing antimicrobial peptides according to claim 4, characterized in that: The dosage form of the perianal care composition includes hydrogel, ointment, spray or film.

7. A method for preparing an anal perianal care composition containing antimicrobial peptides, characterized in that: include: Add glycerin and hyaluronic acid to the swollen carbomer and stir evenly to form a matrix system; Silk fibroin hydrolyzate, phenoxyethanol, cecropin, melittin and scorpion peptide ABP-W1 are added to the matrix system, and the pH value is adjusted to 5.5-6.5 to obtain an anal care composition.

8. The method for preparing the perianal care composition containing antimicrobial peptides according to claim 7, characterized in that: The mixing temperature of the carbomer, glycerin and hyaluronic acid is 45±2° C., and the stirring speed is 300-500 rpm.

9. The method for preparing the perianal care composition containing antimicrobial peptides according to claim 7, characterized in that: After the matrix system is cooled to 25-30° C., silk fibroin hydrolyzate, phenoxyethanol, cecropin, melittin and scorpion peptide ABP-W1 are added.

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