A veterinary enrofloxacin compound gel ear drops and its preparation method and application

Abstract

The present invention belongs to the technical field of veterinary preparations, and specifically relates to a veterinary enrofloxacin compound gel ear drops and its preparation method and application. The compound gel ear drops include four active substances: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone. The preparation method is to add poloxamer, L-cysteine, and carbomer to water, stir and add benzyl alcohol to obtain solution A, dissolve low-erucic acid rapeseed oil in ethanol of more than 85%, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B; slowly add solution B to the gel matrix of solution A to obtain solution C, and then filter to obtain the compound gel. Pharmacodynamic experiments have confirmed that it has a good therapeutic effect on inflammation of the external ear canal of pets; accelerated experiments have confirmed that the compound gel is stable in nature, can be stored for a long time, and is suitable for further expansion of production.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drug preparations and relates to veterinary enrofloxacin compound gel ear drops and a preparation method and application thereof. Background Art

[0002] In my country, pets are mainly dogs and cats. The "2025 China Pet Industry White Paper (Consumption Report)" shows that the number of pets in 2024 will be 124.11 million, an increase of 2.1% over 2023. Among them, the number of pet dogs will be 52.58 million, an increase of 1.6% over 2023, and the number of pet cats will be 71.53 million, an increase of 2.5% over 2023. The average annual consumption of a single pet dog is 2,961 yuan, an increase of 3.0% over 2023. The average annual consumption of a single pet cat is 2,020 yuan, an increase of 4.9% over 2023.

[0003] Numerous reports have been published on the pathogenic bacteria that cause fungal infections of the external auditory canal. In 1988, Stern et al. proposed that Aspergillus, Penicillium citrinum, and Candida albicans were the primary pathogens. In 2007, Yang et al. reported that the most common pathogens of fungal infections of the external auditory canal were Aspergillus niger, Aspergillus fumigatus, Mucor, and Trichophyton mentagrophytes.

[0004] Enrofloxacin has transaminase and topoisomerase IV activity, interfering with bacterial DNA replication, transcription, and repair, thereby achieving antibacterial effects. It is commonly used to treat urinary tract infections, respiratory tract infections, skin and soft tissue infections, etc. The molecular structure is as follows:

[0005]

[0006] Triamcinolone acetonide is a steroid drug belonging to the glucocorticoid class with anti-inflammatory and immunosuppressive properties. It is commonly used to treat a variety of inflammatory diseases, such as allergic diseases, rheumatic autoimmune diseases, and skin diseases. Triamcinolone acetonide can alleviate symptoms by reducing local inflammatory responses and is commonly used for injection or topical application. Due to its immunosuppressive effects, long-term use requires attention to side effects such as osteoporosis and gastrointestinal discomfort. The molecular structure is as follows:

[0007]

[0008] Gentamycin sulfate is an aminoglycoside antibiotic with broad-spectrum antibacterial activity, primarily used to treat serious infections caused by sensitive bacteria, such as sepsis, pneumonia, and peritonitis. It kills bacteria by inhibiting bacterial protein synthesis. Due to the potential for ototoxicity and nephrotoxicity, caution is advised when using it. Blood concentrations and renal function should generally be monitored under a physician's guidance. The molecular structure is as follows:

[0009]

[0010] Pulegone is a monoterpenoid compound extracted from the mint plant and is commonly used as a fragrance, aromatherapy agent, and pharmaceutical ingredient. It has a cooling effect and exhibits certain analgesic, anti-inflammatory, and antibacterial properties. Pulegone is commonly used in ointments, topical medications, and aromatherapy to relieve symptoms such as headaches and indigestion. In some cases, it can also be used as a topical antipruritic. Its molecular structure is as follows:

[0011]

[0012] Currently, the four active substances have not been found in the market to be prepared into gel ear drops, nor have they been used to treat external otitis in pets. Summary of the Invention

[0013] The main purpose of the present invention is to provide a veterinary enrofloxacin compound gel ear drop solution, specifically a compound gel ear drop solution prepared from a combination of four active ingredients: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone. The solution has significant therapeutic effects in treating inflammation, particularly inflammation of the external auditory canal.

[0014] Specifically, the present invention adopts the following technical solutions to achieve the above objectives:

[0015] Disclosed are veterinary enrofloxacin compound gel ear drops, which are composed of the following components in parts by weight: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone in a ratio of 1.2-3.6:0.8-1.2:0.3-0.9:0.1-0.4; and poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol in a ratio of 21-44:1.1-1.4:2.1-4.7:0.3-0.8:1.5-3.5:45-80:13-26.

[0016] The second object of the present invention is to provide a method for preparing the veterinary compound gel ear drops, comprising the following steps:

[0017] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 5-15°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0018] 2) Dissolve rapeseed oil in 80-90% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, and adjust the pH to obtain solution B, which is set aside;

[0019] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 5-15°C, stirring while adding, and maintaining a stirring speed of 150 rpm during the addition process; after the entire solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0020] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0021] Furthermore, the preparation of the veterinary compound gel ear drops comprises the following steps:

[0022] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 5-15°C. Then add benzyl alcohol and stir until completely dissolved to obtain solution A, which is set aside.

[0023] 2) Dissolve rapeseed oil in 80-90% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, and adjust the pH to 4.0-5.0 with citric acid to obtain solution B, which is set aside;

[0024] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 5-15°C, stirring while adding, and always stirring at 150 rpm / min during the addition process; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside.

[0025] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0026] Furthermore, the poloxamer is composed of poloxamer 407 and poloxamer 184, with a weight ratio of 3-6:0.5-0.9; the carbomer is composed of carbomer 934P or carbomer 941, with a weight ratio of 0.2-0.7:5-10.

[0027] Furthermore, the poloxamer is composed of poloxamer 407 and poloxamer 184, with a weight ratio of 5:0.7; the carbomer is composed of carbomer 934P or carbomer 941, with a weight ratio of 0.4:7.3.

[0028] Furthermore, the ratio of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone is 2.4:1:0.6:0.25 respectively; the ratio of poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol is 33:125:3.4:1.05:2:62:19, among which the ratio of poloxamer 407 and poloxamer 184 is 5:0.7, and the ratio of carbomer 934P and carbomer 941 is 0.4:7.3.

[0029] The further poloxamer is composed of poloxamer 407 and poloxamer 184, with a weight ratio of 5:0.7; the said carbopoloxamer 407 and poloxamer 184 are 5:0.7, and the carbomer 934P and carbomer 941 are 0.4:7.3.

[0030] Furthermore, in a preferred embodiment of the present invention, the veterinary enrofloxacin compound gel ear drops have the following ingredients, calculated by weight: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone in a ratio of 2.4:1:0.6:0.25, respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol in a ratio of 33:125:3.4:1.05:2:62:19, wherein poloxamer 407 and poloxamer 184 are in a ratio of 5:0.7, and carbomer 934P and carbomer 941 are in a ratio of 0.4:7.3.

[0031] Furthermore, in a preferred embodiment of the present invention, the veterinary enrofloxacin compound gel ear drops comprises the following steps:

[0032] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 10°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0033] 2) Dissolve rapeseed oil in 85% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, add citric acid to adjust the pH to 4.5, and set aside;

[0034] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 10°C and stirring at 150 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0035] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0036] The veterinary enrofloxacin compound gel ear drops are used in the preparation of a drug for treating inflammation of the external auditory canal of animals. The animals may be poultry, pets, etc., and are particularly suitable for cats and dogs.

[0037] Compared with the prior art, the present invention has the following beneficial effects:

[0038] 1) Enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are mixed in a specific weight ratio in the compound gel of the present invention, and can exert a synergistic effect in pharmacodynamic experiments. Compared with gels prepared by replacing or lacking one active substance, the drug effect is more significant. The compound gel of the present invention has a synergistic antibacterial effect and has a good therapeutic effect on external otitis in animals.

[0039] 2) The compound gel ear drops of the present invention optimize the excipient components and excipient ratios to improve the stability of the compound gel. Accelerated experiments found that the four active ingredients did not significantly decrease. To improve drug permeability, the inventors made creative choices. The selected excipients, such as benzyl alcohol and L-cysteine, can promote the transdermal penetration of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone, thereby exerting a better therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 : There was a significant difference between the normal control group and the model group (P<0.01); there was a significant difference between Examples 1-3 and Comparative Examples 1-2 (P<0.01).

[0041] Figure 2 : Detection of the cumulative permeation of enrofloxacin from 0 to 20 hours.

[0042] Figure 3 : Detection of the cumulative permeation of triamcinolone acetonide from 0 to 20 hours.

[0043] Figure 4 : Detection of the cumulative permeation of gentamicin sulfate from 0 to 20 hours.

[0044] Figure 5 : Detection of the cumulative permeation of pulegone from 0 to 20 hours.

[0045] Figure 6 : Enrofloxacin content at 0, 1, 2, 3, and 6 months.

[0046] Figure 7 : The content of triamcinolone acetonide at 0, 1, 2, 3, and 6 months.

[0047] Figure 8 : The levels of gentamicin sulfate at 0, 1, 2, 3, and 6 months.

[0048] Figure 9 : The levels of pulegone at 0, 1, 2, 3, and 6 months. DETAILED DESCRIPTION

[0049] The present invention is further illustrated by the following examples. It should be understood that the examples are only representative formulations and are intended to illustrate the present invention, not to limit the present invention. Therefore, simple modifications of the present invention based on the method of the present invention fall within the scope of the present invention.

[0050] Example 1: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0051] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0052] The preparation process is:

[0053] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 10°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0054] 2) Dissolve rapeseed oil in 85% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, add citric acid to adjust the pH to 4.5, and set aside;

[0055] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 10°C and stirring at 150 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0056] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0057] Example 2: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0058] Calculated by weight, enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are in the ratio of 1.2:0.8:0.3:0.1; poloxamer, carbomer, benzyl alcohol, L-cysteine, rapeseed oil, water, and ethanol are in the ratio of 21:1.1:2.1:0.3:1.5:45:13; wherein the ratio of poloxamer 407 to poloxamer 184 is 3:0.5, and the ratio of carbomer 934P to carbomer 941 is 0.2:5;

[0059] The preparation process is:

[0060] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 5°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0061] 2) Dissolve rapeseed oil in 80% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, add citric acid to adjust the pH to 4.0, and set aside;

[0062] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 5°C and stirring at 100 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0063] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0064] Example 3: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0065] Calculated by weight, the ratio of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone is 3.6:1.2:0.9:0.4; the ratio of poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol is 44:1.4:4.7:0.8:3.5:80:26; among them, the ratio of poloxamer 407 to poloxamer 184 is 6:0.9, and the ratio of carbomer 934P to carbomer 941 is 0.7:10.

[0066] The preparation process is:

[0067] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 15°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0068] 2) Dissolve rapeseed oil in 90% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, add citric acid to adjust the pH to 5.0, and set aside;

[0069] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 15°C and stirring at 200 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0070] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0071] Comparative Example 1: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0072] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, and gentamicin sulfate are 2.4:1:0.6 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0073] The preparation process is the same as that of Example 1.

[0074] Comparative Example 2: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0075] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, clotrimazole, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0076] The preparation process is the same as that of Example 1.

[0077] Comparative Example 3: A veterinary enrofloxacin compound gel ear drops and a preparation method thereof. The ingredients are calculated by weight: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, propylene glycol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0078] The preparation process is the same as that of Example 1.

[0079] Comparative Example 4: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0080] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, water, and ethanol are 33:125:3.4:1.05:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0081] The preparation process is the same as that of Example 1.

[0082] Comparative Example 5: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0083] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0084] The preparation process is the same as that of Example 1.

[0085] Comparative Example 6: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0086] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, of which the poloxamer is poloxamer 407 and the carbomer is carbomer 941.

[0087] The preparation process is the same as that of Example 1.

[0088] Comparative Example 7: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0089] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 235 and poloxamer 124 are 5:0.7, and carbomer 940 and carbomer 941 are 0.4:7.3.

[0090] The preparation process is the same as that of Example 1.

[0091] Comparative Example 8: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0092] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0093] The preparation process is:

[0094] 1) Add poloxamer and carbomer to water, stir evenly, and stir at 10°C until completely dissolved to obtain Solution A, which is set aside;

[0095] 2) dissolving benzyl alcohol and rapeseed oil in 85% or greater ethanol, adding L-cysteine, enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH to 4.5, and setting aside;

[0096] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 10°C and stirring at 150 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0097] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0098] Comparative Example 9: A veterinary enrofloxacin compound gel ear drops and its preparation method

[0099] Calculated by weight, the ingredients are as follows: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 2.4:1:0.6:0.25 respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 33:125:3.4:1.05:2:62:19, among which poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0100] The preparation process is the same as that of Example 1.

[0101] The preparation process is:

[0102] 1) Add poloxamer, L-cysteine, and carbomer to water, stir evenly, and stir at 10°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A, which is set aside.

[0103] 2) dissolving rapeseed oil in 85% or greater ethanol, and adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B;

[0104] 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 10°C and stirring at 150 rpm / min; after all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside;

[0105] 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

[0106] 1. Pharmacodynamics Experiment

[0107] 1.1 Preparation of fungi

[0108] Preparation of Aspergillus fumigatus mycelial suspension: Take an appropriate amount of Aspergillus fumigatus spores and inoculate them into 250 ml of sterilized Sabourou liquid medium. After culturing in a constant temperature shaking water bath at 37°C for 48 h, centrifuge at a radius of 15 cm and 2000 rpm for 5 min. Discard the supernatant and transfer the precipitate to a 1500 μL EP tube. Grind the mycelium until it is 20-40 μm fragments. Count the number of mycelial fragments with a hemocytometer under an optical microscope and adjust the cell density of the suspension to 1 × 10 9 Spores / ml.

[0109] 1.2 Grouping of experimental animals

[0110] Forty-eight Wister rats were randomly divided into six groups, with eight rats in each group. Both ears were used as experimental subjects. 9 Healthy rats were inoculated with Aspergillus fumigatus spores / ml, and another 8 rats were taken as the normal control group and inoculated with 0.9% sodium chloride solution.

[0111] 1.3 Establishment of a rat model of external otic canal fungal infection

[0112] All rats were anesthetized with 10% chloral hydrate 3ml / kg injected into the abdomen. The external auditory canals of both ears of the rats were selected as the research subjects. Sterile sandpaper was rolled into a rod and gently scratched in an "*" shape to prevent bleeding and damage to the tympanic membrane, causing clinical natural external auditory canal injury, simulating the external auditory canal skin injury caused by natural ear picking in humans. The amount of the prepared Aspergillus fumigatus suspension applied was 200μL each time, and this operation was repeated for 7 days, and the inoculation was completed. The normal control group applied 0.9% sodium chloride solution to the corresponding scratched areas of the external auditory canal.

[0113] 1.4 Administration

[0114] After the model group was established, the drug was administered on the 7th day (corresponding to the examples), with a thickness of about 0.1 mm, for 7 consecutive days.

[0115] 1.5 Detection indicators

[0116] Each example group was given a final score according to the treatment results in Table 1.

[0117] Table 1

[0118]

[0119] 1.4 Points statistical analysis

[0120] Graphpadprism 9.0 software was used to perform statistical analysis on the data. The difference was considered statistically significant when P < 0.05.

[0121] 1.6 Results Analysis

[0122] Figure 1 : "##" There is a significant difference between the normal control group and the model group (P<0.01); for the comparison between Example 1-3 and Comparative Example 1-2, see Figure 1 It can be seen that there are significant differences (P<0.01) between "**" Examples 1-3 and Comparative Examples 1-4.

[0123] 2. Transdermal Diffusion Experiment

[0124] 2.1. Test Drugs

[0125] The compound gel ear drops prepared in Example 1, Comparative Example 1, Comparative Example 3, Comparative Example 5, Comparative Example 6, Comparative Example 7 and Comparative Example 9.

[0126] 2.2 Experimental Methods

[0127] The abdominal skin of 8-week-old rats was peeled off, and the dermis side was used as the receiving layer. Phosphate buffered saline was filled inside the skin, and warm water at 37°C was refluxed in the water jacket. The above-mentioned test drug was punched into a circular shape (1.8 cm 2 ), the patch was applied and the skin was removed, and the receiving fluid was sampled over time. The permeation amount of each drug (enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone) was measured by high performance liquid chromatography to compare the transdermal diffusion of the drugs in different examples.

[0128] 2.3 Cumulative transmittance detection

[0129] See the results Figure 2-5 .

[0130] 2.4 Statistical analysis

[0131] Graphpadprism 9.0 software was used to perform statistical analysis on the data. The difference was considered statistically significant when P < 0.05.

[0132] 2.5 Results Analysis

[0133] Figure 2-5The skin permeation process demonstrates that the compound gel of the present invention, through the careful design and optimization of the interplay of various excipients, forms a highly synergistic system that effectively promotes the penetration of active pharmaceutical ingredients into the skin, thereby significantly increasing their permeation rate. These excipients not only help improve the solubility and stability of the drug but also, through multiple mechanisms, synergistically act on the skin barrier, enhancing the ability of drug molecules to cross the skin. The compound gel prepared by the present invention not only improves the transdermal absorption efficiency of the drug but also ensures that the drug ingredients can exert a stronger therapeutic effect deep within the skin.

[0134] 3. Comparison of stability experiments

[0135] 3.1 Detection method of triamcinolone acetonide

[0136] [Content determination] Determine according to high performance liquid chromatography (General Rule 0512).

[0137] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; methanol-water (525:475) was used as mobile phase; and the detection wavelength was 240 nm.

[0138] Reference solution: Dissolve triamcinolone acetonide in 70% methanol solution and dilute to make a solution containing approximately 5 μg per 1 ml. Take 20 μL and inject it into the liquid chromatograph to record the chromatogram.

[0139] Test solution: Accurately weigh 1.0 g of gel into a 10 mL volumetric flask, dilute to the corresponding scale with pure methanol, shake thoroughly, take 100 μL of solution, add 900 μL of 50% methanol, shake evenly, accurately measure 20 μL and inject into the liquid chromatograph, record the chromatogram.

[0140] 3.2 Detection method of gentamicin sulfate

[0141] [Content determination] Determine according to high performance liquid chromatography (General Rule 0512).

[0142] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel as the filler (pH adaptation range 0.8-8.0); 0.2 mol / L trifluoroacetic acid solution-methanol (96:4) as the mobile phase; the flow rate was 0.6-0.8 ml per minute; and the evaporative light scattering detector (high-temperature splitless mode: drift tube temperature 105-110°C, carrier gas flow rate 2.5 L per minute; low-temperature split mode: drift tube temperature 45-55°C, carrier gas pressure 350 kPa) was used for measurement.

[0143] Reference solution: Take an appropriate amount of gentamicin sulfate standard, dissolve it in the mobile phase and dilute it to make a solution containing approximately 2.5 mg of gentamicin per 1 ml. Take 20 μl of each solution and inject it into the liquid chromatograph to record the chromatogram.

[0144] Test solution: Accurately weigh 1.0 g of gel into a 10 mL volumetric flask, dilute to the corresponding scale with pure methanol, shake thoroughly, take 100 μL of solution, add 900 μL of 50% methanol, shake evenly, accurately measure 20 μL and inject into the liquid chromatograph, record the chromatogram.

[0145] 3.3 Enrofloxacin Detection Method

[0146] [Content determination] Determine according to high performance liquid chromatography (General Rule 0512).

[0147] Chromatographic conditions: Column: Thermos; Mobile phase: Methanol: Water = 60:40; Detection wavelength: 256 nm

[0148] Column temperature: room temperature; flow rate: 1.0 mL / min; injection volume: 20 μL

[0149] Test solution: Accurately weigh 1.0 g of gel into a 10 mL volumetric flask, dilute to the corresponding scale with pure methanol, shake thoroughly, take 100 μL of the solution, add 900 μL of 50% methanol, shake well, and inject for analysis;

[0150] Reference solution: Dilute 0.5 g of enrofloxacin to the appropriate mark with pure methanol. Shake thoroughly. Take 100 μL of this solution and add 900 μL of 50% methanol. Shake thoroughly and determine the solution in the same manner. Calculate the peak area using the external standard method.

[0151] 3.4 Detection method of pulegone

[0152] [Content determination] Determine according to high performance liquid chromatography (General Rule 0512).

[0153] Preparation method of reference solution: accurately weigh pulegone, add chromatographic ethanol to make it contain 0.190 mg of pulegone per ml, shake evenly, and inject for analysis.

[0154] Test solution: Accurately weigh 1.0 g of gel into a 10 mL volumetric flask, dilute to the corresponding scale with chromatographic ethanol, shake thoroughly, take 100 μL of the solution, add 900 μL of 50% methanol, shake well, and inject for analysis.

[0155] Chromatographic determination conditions: octadecylsilane bonded silica gel as filler; acetonitrile mobile phase A, water mobile phase B (mobile phase A: mobile phase B is 10-80:90;20), detection wavelength is 238 nm, flow rate is: 1 ml / min, injection volume is 5 μL.

[0156] 3.5 Testing conditions

[0157] Accelerated Test: The compound gels prepared in the Examples and Comparative Examples were used as test samples and stored at a temperature of 40±2°C and a relative humidity of 75±5% for 6 months. The contents of the four active ingredients were measured at 0, 1, 2, 3, and 6 months, respectively, and calculated using the peak area using the external standard method.

[0158] 3.6 Test results

[0159] pass Figure 6-9 As can be seen, after accelerated testing, the contents of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone in Example 1 were significantly higher than those in Comparative Examples 4-5 and Comparative Examples 8-9, indicating that the use of different excipients or additives, or different processes, significantly impacts the four active ingredients. This demonstrates the rationality, superiority, and creativity of the formulation provided in this application, and the application is expected to have excellent market prospects.

[0160] In summary, the compound gel formulation provided in this application demonstrates a high degree of rationality, superiority, and innovation. Through the precise selection and optimization of the ratios of various excipients, the transdermal absorption of the drug is significantly enhanced while ensuring the stability and efficacy of the drug ingredients. This formulation demonstrates significant advantages in enhancing drug permeability, prolonging the duration of efficacy, and reducing side effects, and has promising market prospects and commercial potential.

Claims

1. A veterinary enrofloxacin compound gel ear drops, characterized in that: The gel ear drops have the following components calculated by weight: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone in a ratio of 2.4:1:0.6:0.25, respectively; poloxamer, carbomer, benzyl alcohol, L-cysteine, rapeseed oil, water, and ethanol in a ratio of 33:125:3.4:1.05:2:62:19, wherein poloxamer 407 and poloxamer 184 are in a ratio of 5:0.7, and carbomer 934P and carbomer 941 are in a ratio of 0.4:7.3; The method for preparing the gel ear drops comprises the following steps: 1) Add poloxamer, L-cysteine, and carbomer to water and stir evenly at 10°C. Then add benzyl alcohol and stir until completely dissolved to obtain Solution A. Set aside. 2) Dissolve rapeseed oil in 85% or greater ethanol, add enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, add citric acid to adjust the pH to 4.5, and set aside; 3) Slowly add Solution B to the gel matrix of Solution A, controlling the temperature to 10°C and stirring at 150 rpm / min. Once all the solution is added, ensure uniform mixing to obtain Solution C, which is set aside. 4) Filter solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

2. Use of the veterinary enrofloxacin compound gel ear drops according to claim 1 in the preparation of a medicament for treating inflammation of the external auditory canal of animals.

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