Compound miconazole nitrate lotion and preparation process thereof

Abstract

The present application relates to the technical field of pharmaceutical formulations, and particularly relates to a compound miconazole nitrate lotion and a preparation process thereof, which comprises the following raw materials in percentage by weight: miconazole nitrate 1.5-2.5%, amphiphilic polymer 5-6%, chlorhexidine gluconate 1.5-2.5%, casone 0.05-0.1%, benzoic acid 0.1-0.2%, surfactant 25-32%, PEG-120 methyl glucose dioleate 1-2%, citric acid monohydrate 0.3-0.4%, and sodium chloride 0.7-0.9%, and the rest is water. The compound miconazole nitrate lotion prepared by the present application has dual effects of anti-fungus and antibiosis, expands the scope of indications, enhances the overall antibacterial effect of the lotion, and is suitable for the treatment and prevention of various skin infections.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, and in particular to a compound miconazole nitrate lotion and its preparation process. Background Technology

[0002] Fungal skin diseases are a group of skin diseases caused by fungal infections. The main pathogens include dermatophytes, Candida, and Malassezia, and are characterized by high incidence and recurrence. Common clinical fungal skin diseases include tinea pedis (athlete's foot), tinea corporis / cruris, and pityriasis versicolor, often presenting as vesicular-scaly lesions, usually localized to one side, and are contagious. Miconazole nitrate is a broad-spectrum antifungal drug. Its mechanism of action mainly involves inhibiting ergosterol synthase on the fungal cell membrane, preventing ergosterol synthesis, thereby disrupting the structure and function of the fungal cell membrane and leading to fungal death.

[0003] While miconazole nitrate performs well in treating fungal skin diseases, its efficacy is limited in mixed infections or skin conditions with inflammatory responses. Mixed infections often involve multiple pathogens, including bacteria and fungi, and miconazole nitrate is effective only against fungi, not bacteria. Furthermore, skin conditions with inflammatory responses may require both suppressing the inflammatory response and killing the pathogens, and miconazole nitrate does not possess anti-inflammatory properties. Therefore, in the treatment of mixed infections or skin conditions with inflammatory responses, the use of miconazole nitrate alone may not achieve the desired therapeutic effect.

[0004] Chinese patent application CN105079159A discloses a metronidazole and miconazole nitrate compound lotion and its preparation method. The lotion comprises 5-10 parts metronidazole, 8-12 parts miconazole nitrate, 25-30 parts Corydalis yanhusuo, 45-50 parts catechu, 35-40 parts whitefish tail, 35-40 parts nine-head grass, 50-55 parts Polygonum multiflorum, 25-30 parts Portulaca oleracea, 50-55 parts dog ant grass, 35-40 parts prepared licorice root, 15-20 parts Angelica dahurica, and 8-12 parts cicada slough. This lotion is a combination of traditional Chinese and Western medicine, which not only improves the bactericidal / bacteriostatic spectrum but also overcomes the limitation of Western medicine primarily inhibiting the growth of most fungi. However, it cannot quickly relieve patient discomfort for inflammatory symptoms such as skin erythema, itching, and exudation associated with fungal infections (e.g., seborrheic dermatitis or eczema with infection), leading to a prolonged treatment period and decreased patient compliance due to persistent symptoms. Meanwhile, this wash is a compound of traditional Chinese medicine, which takes a long time to indirectly reduce inflammation through its antibacterial effect. However, patients have a strong need for immediate relief of symptoms such as itching and pain, which may lead to discontinuation of the medication during treatment and affect its efficacy. Summary of the Invention

[0005] To address the problems mentioned in the background section, this invention provides a compound miconazole nitrate lotion and its preparation process.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A compound miconazole nitrate lotion comprises, by weight percentage, the following raw materials: miconazole nitrate 1.5-2.5%, amphiphilic polymer 5-6%, chlorhexidine gluconate 1.5-2.5%, Kathon 0.05-0.1%, benzoic acid 0.1-0.2%, surfactant 25-32%, PEG-120 methyl glucoside ester 1-2%, citric acid monohydrate 0.3-0.4%, and sodium chloride 0.7-0.9%, with the balance being water.

[0008] Furthermore, the surfactant includes one or more of lauryl polyoxyethylene ether, cocamidopropyl betaine, disodium cocoamphodiacetate cetyltrimethylammonium chloride, sodium dodecyl sulfate, polysorbate-80, cocamidopropyl hydroxysulfonate betaine, decyl glucoside, and sodium lauroyl sarcosinate.

[0009] Furthermore, the amphiphilic polymer is prepared by the following steps:

[0010] S1. Under nitrogen protection, glycerol and sodium hydroxide are mixed and heated for condensation reaction for 2-3 hours to obtain a pale yellow viscous liquid polyglycerol. Polyglycerol, ricinoleic acid and n-hexane are mixed evenly and placed in a microwave reactor for 10-20 minutes. Immobilized lipase and zeolite molecular sieve are added, and the reaction is continued at a constant temperature with shaking for 12-14 hours. After the reaction is completed, the lipase is recovered by filtration through a 200-mesh sieve. Distillation is carried out at 80℃ / 10Pa to remove unreacted substances. The main fraction is collected at 155℃ / 0.1Pa. Silica gel column chromatography (silica gel 60) is used with ethyl acetate / petroleum ether = 1:5 as eluent to remove pigments and trace amounts of free acid. A colorless and transparent oily substance is collected as polyglycerol ricinoleic acid ester.

[0011] S2. Add polyglycerol ricinoleate and gallic acid to the reactor, add N-methylpyrrolidone, and stir under reflux for 1-2 hours to remove moisture from the raw materials. Add p-toluenesulfonic acid to the reaction system, raise the temperature and continue the reaction for 5-6 hours. After the reaction is completed, cool the reaction solution to 60°C, add pre-cooled anhydrous ethanol, filter using a vacuum filter, wash three times with a mixture of ether / ethanol (1:3), place in a vacuum drying oven and dry at 40°C for 24 hours to obtain a white waxy solid.

[0012] Further, in step S1, the mass ratio of glycerol to sodium hydroxide is (190-210):(0.95-1.05), and the mass ratio of polyglycerol, ricinoleic acid, n-hexane, lipase and zeolite molecular sieve is (20-22):(44-50):(40-41):1:(6-7).

[0013] Furthermore, in step S1, the heating temperature is 230-240℃, the microwave reaction temperature is 50-60℃, the power is 200-300W, and the frequency is 2.45-2.55GHz.

[0014] Furthermore, in step S1, the temperature of the isothermal oscillation is 60-65℃, and the speed is 200-300rpm.

[0015] Further, in step S2, the mass ratio of polyglycerol ricinoleate, gallic acid, N-methylpyrrolidone and p-toluenesulfonic acid is (33-35):(6-7):(82-84):(1-1.65).

[0016] Furthermore, in step S2, the temperature is controlled at 80-85℃, the stirring speed is 100-200rpm, and the heating temperature is 110-115℃.

[0017] According to another aspect of the present invention, the preparation process of the above-mentioned compound miconazole nitrate lotion includes the following steps:

[0018] Add water to a mixer, heat, add PEG-120 methyl glucoside ester and surfactant, stir for 30-40 minutes until completely dissolved, add miconazole nitrate, chlorhexidine gluconate, amphiphilic polymer, Kathon and benzoic acid in sequence, continue stirring for 15-25 minutes, add citric acid monohydrate to adjust the pH value, then add sodium chloride, homogenize for 10-20 minutes, filter to remove impurities, and fill to obtain the finished product.

[0019] Furthermore, the heating temperature is 40-45℃, the stirring speed is 500-600 rpm, and the homogenization speed is 2000-3000 rpm.

[0020] Beneficial effects of this invention:

[0021] 1. In the technical solution of this invention, under alkaline conditions and nitrogen protection, the hydroxyl groups of glycerol undergo a dehydration condensation reaction, followed by ring-opening polymerization via nucleophilic substitution. The primary hydroxyl group of glycerol attacks the secondary hydroxyl oxygen atom of an adjacent glycerol molecule, removing water molecules and forming an ether bond to generate linear or branched polyglycerol. The polyglycerol then undergoes regioselective transesterification with ricinoleic acid under the catalysis of an immobilized lipase. The serine residue of the lipase attacks the carboxyl carbon of ricinoleic acid, forming an acyl-enzyme intermediate. Subsequently, the hydroxyl oxygen of the polyglycerol nucleophilically attacks the acyl carbon, completing the ester bond construction. Polyglycerol ricinoleic acid is further esterified and grafted with gallic acid. p-Toluenesulfonic acid protonates the hydroxyl group of polyglycerol ricinoleic acid or the carboxyl group of gallic acid, forming a more easily leaving group. The carboxyl carbon of gallic acid is nucleophilically attacked by the hydroxyl oxygen of polyglycerol ricinoleic acid, generating a phenolic ester bond, which is grafted onto the polyglycerol ricinoleic acid backbone. The benzene ring structure of gallic acid enhances hydrophobic interactions through π-π stacking, while its three phenolic hydroxyl groups further strengthen hydrophilicity. The rigid planar structure of gallic acid restricts the conformational freedom of the polymer chain, promoting the formation of stable micelles or layered structures. The resulting polymer exhibits amphiphilicity; the polyglycerol segments form a hydrogen-bonded network of hydrophilic regions with the phenolic hydroxyl groups of gallic acid, while the ricinoleic acid chains form hydrophobic regions with the benzene ring of gallic acid through hydrophobic association. Because both hydrophilic and hydrophobic segments exist simultaneously in the polymer molecule, microphase separation occurs in selective solvents, spontaneously forming self-assembled structures such as spherical, rod-shaped, and layered structures. In compound miconazole nitrate lotion, this self-assembly behavior helps form a stable suspension or solution system, encapsulating and stabilizing various components in the lotion, preventing separation or precipitation due to incompatibility, thereby improving the stability and uniformity of the lotion.

[0022] 2. In the technical solution of the present invention, the amphiphilic polymer has a significant solubilizing effect. Its hydrophobic segments can interact with miconazole nitrate in the lotion, and increase the solubility of these components in water by forming stable micelles or solutions. This is beneficial to improving the activity and effect of the lotion, ensuring that the effective components in the lotion can be fully dissolved and evenly distributed in the lotion, thereby acting more effectively on the skin or mucous membrane surface.

[0023] 3. In the technical solution of this invention, the amphiphilic polymer also exhibits good emulsifying properties. Its hydrophilic and hydrophobic segments can interact with the aqueous and oil phases respectively, mixing incompatible liquids or solids uniformly to form a stable suspension or solution. In compound miconazole nitrate lotion, it helps to uniformly disperse various components (including aqueous and oily components) in water, forming a stable lotion system and improving the ease of use of the lotion.

[0024] 4. In the technical solution of this invention, miconazole nitrate targets fungal infections, while chlorhexidine gluconate, as a broad-spectrum antibacterial agent, effectively inhibits bacteria. The combination of the two achieves dual antifungal and antibacterial effects, expands the scope of indications, enhances the overall antibacterial effect of the lotion, and is suitable for the treatment and prevention of various skin infections.

[0025] 5. In the technical solution of this invention, the compound surfactant system balances cleansing power and gentleness, reduces damage to the skin barrier, and is suitable for sensitive skin and long-term use. The combined use of Kathon and benzoic acid achieves highly effective preservation at low concentrations, reducing the risk of allergies associated with traditional preservatives. Detailed Implementation

[0026] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] Unless otherwise specified, the raw materials used in this invention are all from commercially available conventional products, and the immobilized lipase is prepared using the method described in CN106929502A.

[0028] Preparation Example 1

[0029] Amphiphilic polymers are prepared by the following steps:

[0030] S1. Under nitrogen protection, 190g of glycerol and 0.95g of sodium hydroxide were mixed and heated to 230℃ for condensation reaction for 2h to obtain a pale yellow viscous liquid polyglycerol. 20g of polyglycerol, 44g of ricinoleic acid and 40g of n-hexane were mixed evenly and placed in a microwave reactor. The mixture was treated at 50℃ for 10min. The microwave power was 200W and the frequency was 2.45GHz. 1g of immobilized lipase and 6g of zeolite molecular sieve were added. The mixture was then treated at 60℃ with constant temperature shaking at 200rpm for 12h. After the reaction was completed, the lipase was recovered by filtration through a 200-mesh sieve. The mixture was distilled at 80℃ / 10Pa to remove unreacted substances. The main fraction was collected at 155℃ / 0.1Pa. Silica gel column chromatography (silica gel 60) was used with ethyl acetate / petroleum ether = 1:5 as the eluent to remove pigments and trace amounts of free acid. A colorless and transparent oily substance was collected as polyglycerol ricinoleic acid ester.

[0031] S2. Add 33g of polyglycerol ricinoleate and 6g of gallic acid to the reactor, add 82g of N-methylpyrrolidone, control the temperature at 80℃ and stir and reflux at 100rpm for 1h to remove the moisture from the raw materials, add 1g of p-toluenesulfonic acid to the reaction system, raise the temperature to 110℃ and continue to react for 5h. After the reaction is completed, cool the reaction solution to 60℃, add pre-cooled anhydrous ethanol, filter using a vacuum filter, wash three times with a mixture of ether / ethanol (1:3), place in a vacuum drying oven and dry at 40℃ for 24h to obtain a white waxy solid.

[0032] Preparation Example 2

[0033] Amphiphilic polymers are prepared by the following steps:

[0034] S1. Under nitrogen protection, 200g of glycerol and 1g of sodium hydroxide were mixed and heated to 235℃ for condensation reaction for 2.5h to obtain a pale yellow viscous liquid polyglycerol. 21g of polyglycerol, 46g of ricinoleic acid and 40.5g of n-hexane were mixed evenly and placed in a microwave reactor. The mixture was treated at 55℃ for 15min. The microwave power was 250W and the frequency was 2.5GHz. 1g of immobilized lipase and 6.5g of zeolite molecular sieve were added. The mixture was then treated at 62℃ with constant temperature shaking at 250rpm for 13h. After the reaction was completed, the lipase was recovered by filtration through a 200-mesh sieve. The mixture was distilled at 80℃ / 10Pa to remove unreacted substances. The main fraction was collected at 155℃ / 0.1Pa. Silica gel column chromatography (silica gel 60) was used with ethyl acetate / petroleum ether = 1:5 as the eluent to remove pigments and trace amounts of free acid. A colorless and transparent oily substance was collected as polyglycerol ricinoleic acid ester.

[0035] S2. Add 34g of polyglycerol ricinoleate and 6.5g of gallic acid to the reactor, add 83g of N-methylpyrrolidone, control the temperature at 82℃ and stir and reflux at 150rpm for 1.5h to remove the moisture from the raw materials, add 1.3g of p-toluenesulfonic acid to the reaction system, raise the temperature to 112℃ and continue to react for 5.5h. After the reaction is completed, cool the reaction solution to 60℃, add pre-cooled anhydrous ethanol, filter using a vacuum filter, wash three times with a mixture of ether / ethanol (1:3), place in a vacuum drying oven and dry at 40℃ for 24h to obtain a white waxy solid.

[0036] Preparation Example 3

[0037] Amphiphilic polymers are prepared by the following steps:

[0038] S1. Under nitrogen protection, 210g of glycerol and 1.05g of sodium hydroxide were mixed and heated to 240℃ for condensation reaction for 3h to obtain a pale yellow viscous liquid polyglycerol. 22g of polyglycerol, 50g of ricinoleic acid and 41g of n-hexane were mixed evenly and placed in a microwave reactor. The mixture was treated at 60℃ for 20min. The microwave power was 300W and the frequency was 2.55GHz. 1g of immobilized lipase and 7g of zeolite molecular sieve were added. The mixture was then treated at 65℃ with constant temperature shaking at 300rpm for 14h. After the reaction was completed, the lipase was recovered by filtration through a 200-mesh sieve. The mixture was distilled at 80℃ / 10Pa to remove unreacted substances. The main fraction was collected at 155℃ / 0.1Pa. Silica gel column chromatography (silica gel 60) was used with ethyl acetate / petroleum ether = 1:5 as the eluent to remove pigments and trace amounts of free acid. The resulting colorless and transparent oily substance was polyglycerol ricinoleate.

[0039] S2. Add 35g of polyglycerol ricinoleate and 7g of gallic acid to the reactor, add 84g of N-methylpyrrolidone, control the temperature at 85℃ and stir and reflux at 200rpm for 2h to remove the moisture from the raw materials, add 1.65g of p-toluenesulfonic acid to the reaction system, raise the temperature to 115℃ and continue to react for 6h. After the reaction is completed, cool the reaction solution to 60℃, add pre-cooled anhydrous ethanol, filter using a vacuum filter, wash three times with a mixture of ether / ethanol (1:3), place in a vacuum drying oven and dry at 40℃ for 24h to obtain a white waxy solid.

[0040] Example 1

[0041] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0042] Add 59.57g of water to a mixer and heat to 40°C. Add 1.5g of PEG-120 methyl glucoside dioleate and 28g of surfactant, including 6g of lauryl polyoxyethylene ether, 13g of cocamidopropyl betaine, 8.8g of disodium cocoamphodiacetate and 0.2g of hexadecyltrimethylammonium chloride. Stir at 500rpm for 30min until completely dissolved. Add 2g of miconazole nitrate, 2g of chlorhexidine gluconate, 5.5g of the amphiphilic polymer prepared in Preparation Example 1, 0.08g of Kathon, and 0.15g of benzoic acid. Continue stirring at 500rpm for 15min. Add 0.35g of citric acid monohydrate to adjust the pH value, and then add 0.8g of sodium chloride. Homogenize at 2000rpm for 10min. Filter to remove impurities and fill to obtain the finished product.

[0043] Example 2

[0044] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0045] Add 60.07g of water to a mixer and heat to 40-45℃. Add 1.2g of PEG-120 methyl glucoside dioleate and 30g of surfactant, including 8g of lauryl polyoxyethylene ether, 14g of cocamidopropyl betaine, 7.1g of disodium cocoamphodiacetate and 0.9g of hexadecyltrimethylammonium chloride. Stir at 550rpm for 35min until completely dissolved. Add 1.8g of miconazole nitrate, 1.7g of chlorhexidine gluconate, 5g of the amphiphilic polymer prepared in Preparation Example 2, 0.06g of Kathon, and 0.12g of benzoic acid. Continue stirring at 550rpm for 20min. Add 0.32g of citric acid monohydrate to adjust the pH value, and then add 0.75g of sodium chloride. Homogenize at 2500rpm for 15min, filter to remove impurities, and fill to obtain the finished product.

[0046] Example 3

[0047] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0048] Add 59.13g of water to a mixer and heat to 45°C. Add 1.8g of PEG-120 methyl glucoside dioleate and 26g of surfactant, including 10g of lauryl polyoxyethylene ether, 16g of cocamidopropyl betaine, 9.8g of sodium dodecyl sulfate, and 0.2g of sodium lauroyl sarcosinate. Stir at 600rpm for 40min until completely dissolved. Add 2.3g of miconazole nitrate, 2.2g of chlorhexidine gluconate, 6g of the amphiphilic polymer prepared in Preparation Example 3, 0.09g of Kathon, and 0.18g of benzoic acid. Continue stirring at 600rpm for 25min. Add 0.38g of citric acid monohydrate to adjust the pH value, and then add 0.85g of sodium chloride. Homogenize at 3000rpm for 20min, filter to remove impurities, and fill to obtain the finished product.

[0049] Example 4

[0050] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0051] Add 61.16g of water to a mixer and heat to 40°C. Add 1.6g of PEG-120 methyl glucoside dioleate and 27g of surfactant, including 3g of lauryl polyoxyethylene ether, 16g of cocamidopropyl betaine, 6.6g of polysorbate-80, and 1.4g of hexadecyltrimethylammonium chloride. Stir at 500rpm for 30min until completely dissolved. Add 1.7g of miconazole nitrate, 1.9g of chlorhexidine gluconate, 5.2g of the amphiphilic polymer prepared in Preparation Example 1, 0.07g of Kathon, and 0.13g of benzoic acid. Continue stirring at 500rpm for 15min. Add 0.33g of citric acid monohydrate to adjust the pH value, and then add 0.82g of sodium chloride. Homogenize at 2000rpm for 10min, filter to remove impurities, and fill to obtain the finished product.

[0052] Example 5

[0053] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0054] Add 58.21g of water to a mixer and heat to 40-45℃. Add 1.3g of PEG-120 methyl glucoside dioleate and 29g of surfactant, including 7g of lauryl polyoxyethylene ether, 11.5g of disodium cocoamphodiacetate, 8g of cocamidopropyl hydroxysulfonate, and 2.5g of hexadecyltrimethylammonium chloride. Stir at 550rpm for 35min until completely dissolved. Add 2.2g of miconazole nitrate, 2.1g of chlorhexidine gluconate, 5.8g of the amphiphilic polymer prepared in Preparation Example 2, 0.06g of Kathon, and 0.15g of benzoic acid. Continue stirring at 550rpm for 20min. Add 0.37g of citric acid monohydrate to adjust the pH value, and then add 0.78g of sodium chloride. Homogenize at 2500rpm for 15min, filter to remove impurities, and fill to obtain the finished product.

[0055] Example 6

[0056] The preparation process of compound miconazole nitrate lotion includes the following steps:

[0057] Add 57g of water to a mixer and heat to 45°C. Add 2g of PEG-120 methyl glucoside dioleate and 30g of surfactant, including 10g of cocamidopropyl betaine, 16g of lauryl polyoxyethylene ether, 9.8g of disodium cocoamphodiacetate and 0.2g of hexadecyltrimethylammonium chloride. Stir at 600rpm for 40min until completely dissolved. Add 2g of miconazole nitrate, 2.5g of chlorhexidine gluconate, 5g of the amphiphilic polymer prepared in Preparation Example 3, 0.1g of Kathon, and 0.1g of benzoic acid. Continue stirring at 600rpm for 25min. Add 0.4g of citric acid monohydrate to adjust the pH value, and then add 0.9g of sodium chloride. Homogenize at 3000rpm for 20min. Filter to remove impurities and fill to obtain the finished product.

[0058] Comparative Example 1

[0059] The difference between this comparative example and Example 1 is that polyglycerol is used instead of the amphiphilic polymer prepared in Example 1, while the remaining steps are the same as in Example 1.

[0060] Comparative Example 2

[0061] The difference between this comparative example and Example 2 is that ricinoleic acid was used instead of the amphiphilic polymer prepared in Example 2, while the remaining steps were the same as in Example 2.

[0062] Comparative Example 3

[0063] The difference between this comparative example and Example 3 is that gallic acid was used instead of the amphiphilic polymer prepared in Example 3, while the remaining steps were the same as in Example 3.

[0064] (I) Antibacterial activity test

[0065] Fungal group: Candida albicans and Trichophyton rubrum were inoculated onto Sabouraud dextrose agar plates and cultured at 35°C for 48 h. Single colonies were picked and transferred to 5 mL of RPMI 1640 medium and cultured at 35°C with shaking for 24 h. The bacterial suspension was centrifuged at 3000 rpm for 10 min, the supernatant was discarded, and the turbidity was adjusted to 1 × 10⁻⁶ with sterile physiological saline. 6 CFU / mL, then diluted to 1×10 3 CFU / mL.

[0066] Bacterial group: Staphylococcus aureus was inoculated into MHB, cultured at 35°C with shaking for 18 h, and the turbidity was adjusted to 1×10 after centrifugation. 6 CFU / mL, diluted to 1×10⁻⁶ 3 CFU / mL.

[0067] Take 1g of the compound lotion prepared in Example 1 and Comparative Example 1, dissolve it in 9mL of sterile PBS, vortex mix well, and prepare 10% (w / v) sample stock solution and comparative example stock solution.

[0068] The stock solution of the detergent was serially diluted 2-fold using RPMI 1640 or MHB, with concentration gradients of 64, 32, 16, 8, 4, 2, 1, and 0.5 μg / mL (miconazole nitrate equivalent), to obtain the gradient diluted detergents of the examples and the comparative examples.

[0069] Microdilution plates were used for sample loading, with 200 μL added to each well for both fungal and bacterial groups. Experimental groups, control groups, positive controls, negative controls, and growth controls were included. The sample loading protocol is as follows:

[0070] Experimental group: Each column is set with a concentration gradient, and each concentration is repeated in 3 wells. 100 μL of the gradient dilution wash solution and 100 μL of bacterial suspension are added.

[0071] Control group: Each column is set with a concentration gradient, and each concentration is repeated in 3 wells. 100 μL of comparative gradient dilution washing agent and 100 μL of bacterial suspension are added.

[0072] Positive control: Add 100 μL of pure miconazole nitrate solution (with the same concentration gradient) and 100 μL of bacterial suspension.

[0073] Negative control: Add 200 μL of sterile culture medium.

[0074] Growth control: Add 100 μL of bacterial suspension and 100 μL of culture medium.

[0075] Use a multichannel pipette to add the diluted sample and bacterial suspension sequentially, avoiding air bubbles. Seal the plate surface with a sealing film and incubate at 35°C (48h for fungi, 24h for bacteria). Measure the OD after incubation.

[0076] Fungi: Observe whether the bottom of the well is clear and free of sediment. The minimum concentration that completely inhibits the germination of hyphae or spores is the MIC.

[0077] Bacteria: Observe whether there is no turbidity in the well (consistent with the negative control), and use the lowest concentration without turbidity as the MIC.

[0078] The inhibition rate was calculated as 1 - OD experimental group / OD growth control, with the lowest concentration showing an inhibition rate ≥ 90% defined as the MIC. The results are shown in Table 1.

[0079] Table 1. Results of antibacterial activity test (Minimum inhibitory concentration, MIC, μg / mL)

[0080]

[0081]

[0082] (II) Stability Test

[0083] The composite detergents (20 mL) prepared in Examples 1-6 and Comparative Examples 1-3 were sealed and randomly grouped, with each group numbered and its initial state recorded. They were stored in a constant temperature and humidity chamber (Binder KBF-240) at 40±2℃ and 75±5%RH, protected from light to prevent photodegradation. Samples were taken out after one month for testing, and changes in appearance, miconazole nitrate content, and pH value were recorded. The results are shown in Table 2.

[0084] Table 2. Stability test results of Examples 1-6 and Comparative Examples 1-3

[0085] sample Appearance (color / transparency) Miconazole nitrate content (%) pH value Example 1 Colorless and transparent 99.3 5.1 Example 2 Colorless and transparent 99.5 5.2 Example 3 Colorless and transparent 99.8 5.1 Example 4 Slightly cloudy 98.1 5.3 Example 5 Colorless and transparent 99.0 5.1 Example 6 Colorless and transparent 99.9 5.0 Comparative Example 1 pale yellow and cloudy 94.2 5.2 Comparative Example 2 pale yellow and cloudy 96.8 5.0 Comparative Example 3 pale yellow and cloudy 91.5 4.8

[0086] (III) Skin compatibility assessment

[0087] Take 10 mL of fresh rabbit whole blood and mix it with 2 mL of Alsever's solution. Store at 4°C for no more than 72 hours. Wash the red blood cells by centrifugation at 2000 rpm for 10 min, discarding the supernatant and white blood cell layer. Wash the precipitated red blood cells three times with PBS, centrifuging at 2000 rpm for 10 min each time. Take 1 mL of the washed red blood cells, add 49 mL of PBS, mix well, and then perform a blood cell count, adjusting the result to 2 × 10⁻⁶ cells / mL. 8 cells / mL (≈2% suspension).

[0088] Take 1 mL of each of the detergent samples from Example 1 and Comparative Example 1, add 9 mL of PBS, and vortex mix to a concentration of 10% to obtain a 10% detergent.

[0089] Add each incubation system to a 5 mL centrifuge tube. The incubation system construction scheme is as follows:

[0090] Experimental group: 1 mL of 10% of the lotion from Example 1 + 1 mL of 2% red blood cell suspension.

[0091] Control group: 1 mL of 10% lotion from Comparative Example 1 + 1 mL of 2% red blood cell suspension.

[0092] Positive control: 1 mL of 1% Triton X-100 + 1 mL of 2% red blood cell suspension.

[0093] Negative control: 1 mL physiological saline + 1 mL 2% red blood cell suspension.

[0094] Blank group: 1 mL PBS + 1 mL 2% red blood cell suspension.

[0095] Vortex each group thoroughly, incubate at 37°C in a water bath for 1 hour, then immediately incubate on ice for 5 minutes. Centrifuge at 2000 rpm for 10 minutes at 4°C and collect the supernatant. Transfer 200 μL of the supernatant to a 96-well plate, avoiding aspiration of the precipitate. Measure the absorbance at 540 nm using a microplate reader (zeroing with the blank group). Calculate the hemolysis rate (%): Hemolysis rate (%) = (OD0)0 测试组 -OD 阴性对照 ) / (OD 阳性对照 -OD 阴性对照 )×100. The results are shown in Table 3:

[0096] Table 3. Results of the erythrocyte hemolysis test

[0097] sample <![CDATA[Absorbance (OD 540 )]]> Hemolysis rate (%) Mean ± SD Example 1 0.12,0.11,0.13 4.8％ 4.7±0.5％ Comparative Example 1 0.45,0.47,0.46 58.3％ 58.0±1.2％ Positive control 0.78,0.79,0.77 100％ - Positive control 0.08,0.07,0.08 0％ -

[0098] As shown in Table 1, the antibacterial activity of Example 1 was significantly better than that of Comparative Example 1 and the positive control. This indicates that the micelle structure formed by the amphiphilic polymer encapsulates the hydrophobic drug miconazole nitrate, improving its water solubility and thus increasing the contact area with the microbial cell membrane. The hydrophobic chain of ricinoleate is similar to the lipid layer of the fungal cell membrane (rich in ergosterol), which may promote drug transmembrane penetration. The phenolic hydroxyl group of gallic acid disrupts the integrity of the microbial membrane through hydrogen bonds, synergistically enhancing the effect with the ergosterol synthesis inhibition mechanism of miconazole nitrate. In contrast, Comparative Example 1 may have lacked esterified hydrophobic groups, failing to form effective micelles, resulting in poor drug dispersion and an increased MIC.

[0099] Table 2 shows that the micellar structure of the polymer isolates miconazole nitrate, reducing its direct contact with moisture and oxygen, and inhibiting hydrolysis and oxidative degradation (the content in Comparative Example 1 decreased to 94.2%). Example 1 remained colorless, while Comparative Example 3 became cloudy due to the oxidation of gallic acid to quinones, indicating that the phenolic hydroxyl groups of gallic acid may scavenge free radicals and delay drug oxidation. The pH of Example 1 decreased by only 0.1, while the pH of Comparative Example 3 decreased by 0.4, suggesting that the hydroxyl and carboxylic acid groups in the amphiphilic polymer may stabilize the system pH through hydrogen bonding.

[0100] As shown in Table 3, the hemolysis rate in Example 1 was only 4.7%, indicating that the polymer micelles encapsulate the ionic surfactant, reducing its direct contact with the erythrocyte membrane and lowering the risk of membrane damage. In contrast, Control Group 1, lacking a shielding structure, exhibited a hemolysis rate as high as 58%, demonstrating that the hydrophobic core of polyglycerol ricinoleate adsorbs free drugs and surfactants, while the hydrophilic shell (polyglycerol chain) forms a hydration layer, shielding against stimulation of erythrocytes. The zwitterionic properties of the amphiphilic polymer may neutralize the charge of the surfactant, reducing electrostatic damage to the cell membrane.

[0101] In summary, in Examples 1-6, the hydrophobic core of the amphiphilic polymer, composed of castor oil ester, may have the effect of solubilizing drugs and promoting membrane permeability, while the hydrophilic shell of polyglycerol may improve water solubility and shield irritating components. Through a synergistic effect of multiple mechanisms, the amphiphilic polymer achieves highly efficient antibacterial activity, stable storage, and low irritation.

[0102] In the description of this specification, the terms "preparation example," "example," "various examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that example or preparation example, which are included in at least one example or preparation example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same example or preparation example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more examples or preparation examples.

[0103] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A compound miconazole nitrate lotion, characterized in that, The ingredients, by weight percentage, include: miconazole nitrate 1.5-2.5%, amphiphilic polymer 5-6%, chlorhexidine gluconate 1.5-2.5%, Kathon 0.05-0.1%, benzoic acid 0.1-0.2%, surfactant 25-32%, PEG-120 methyl gluconate dioleate 1-2%, citric acid monohydrate 0.3-0.4%, and sodium chloride 0.7-0.9%, with the balance being water; The amphiphilic polymer is prepared by the following steps: S1. Under nitrogen protection, glycerol and sodium hydroxide are mixed and heated for 2-3 hours to obtain a pale yellow viscous liquid polyglycerol. Polyglycerol, ricinoleic acid and n-hexane are mixed evenly and placed in a microwave reactor for 10-20 minutes. Immobilized lipase and zeolite molecular sieve are added, and the reaction is continued at a constant temperature with shaking for 12-14 hours. After the reaction is completed, the mixture is filtered, distilled twice, and purified by column chromatography to obtain a colorless and transparent oily substance, which is polyglycerol ricinoleic acid ester. S2. Add polyglycerol ricinoleate and gallic acid to the reactor, add N-methylpyrrolidone, control the temperature and stir under reflux for 1-2 hours to remove the moisture from the raw materials, add p-toluenesulfonic acid to the reaction system, raise the temperature and continue the reaction for 5-6 hours. After the reaction is completed, cool to 60°C, filter, wash and dry to obtain a white waxy solid which is an amphiphilic polymer. In step S1, the mass ratio of glycerol to sodium hydroxide is (190-210):(0.95-1.05), and the mass ratio of polyglycerol, ricinoleic acid, n-hexane, lipase and zeolite molecular sieve is (20-22):(44-50):(40-41):1:(6-7). In step S2, the mass ratio of polyglycerol ricinoleate, gallic acid, N-methylpyrrolidone, and p-toluenesulfonic acid is (33-35):(6-7):(82-84):(1-1.65). In step S1, the temperature of the isothermal oscillation is 60-65℃, and the speed is 200-300 rpm; In step S2, the temperature is controlled at 80-85℃, the stirring speed is 100-200 rpm, and the heating temperature is 110-115℃. In step S1, the heating temperature is 230-240℃, the microwave reaction temperature is 50-60℃, the power is 200-300W, and the frequency is 2.45-2.55GHz.

2. The compound miconazole nitrate lotion according to claim 1, characterized in that, Surfactants include one or more of lauryl polyoxyethylene ether, cocamidopropyl betaine, disodium cocoamphodiacetate cetyltrimethylammonium chloride, sodium dodecyl sulfate, polysorbate-80, cocamidopropyl hydroxysulfonate betaine, decyl glucoside, and sodium lauroyl sarcosinate.

3. A preparation process for the compound miconazole nitrate lotion as described in any one of claims 1-2, characterized in that, Includes the following steps: Add water to a mixer, heat, add PEG-120 methyl glucoside ester and surfactant, stir for 30-40 minutes, add miconazole nitrate, chlorhexidine gluconate, amphiphilic polymer, Kathon and benzoic acid in sequence, continue stirring for 15-25 minutes, add citric acid monohydrate, then add sodium chloride, homogenize for 10-20 minutes, filter to remove impurities, and fill to obtain the finished product.

4. The preparation process of the compound miconazole nitrate lotion according to claim 3, characterized in that, The heating temperature is 40-45℃, the stirring speed is 500-600 rpm, and the homogenization speed is 2000-3000 rpm.

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