Veterinary enrofloxacin compound gel ear drops as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of veterinary drug preparations, and particularly relates to veterinary enrofloxacin compound gel ear drops as well as a preparation method and application thereof. The compound gel ear drops comprise four active substances, namely enrofloxacin, triamcinolone acetonide, gentamicin sulfate and pulegone, and the preparation method comprises the following steps: adding poloxamer, L-cysteine and carbomer into water, stirring, adding benzyl alcohol to obtain a solution A, dissolving low-erucic-acid rapeseed oil into ethanol with the concentration of 85% or above, stirring, adding benzyl alcohol into the solution A, and uniformly mixing to obtain the compound gel ear drops. Adding enrofloxacin, triamcinolone acetonide, pulegone and gentamicin sulfate to obtain a solution B; slowly adding the solution B into the solution A gel matrix to obtain a solution C, and then filtering to obtain the compound gel. Pharmacodynamic experiments prove that the traditional Chinese medicine composition has a good treatment effect on external auditory canal inflammation of pets; accelerated experiments prove that the compound gel is stable in property, can be stored for a long time and is suitable for further expanded 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] Pets in my country are mainly dogs and cats. The "White Paper on China's Pet Industry in 2025 (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 is 52.58 million, an increase of 1.6% over 2023, and the number of pet cats is 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] There have been many reports on the pathogenic bacteria of external auditory canal fungal diseases. In 1988, Stern JC et al. proposed that the pathogenic bacteria detected in external auditory canal fungal diseases are mainly Aspergillus, Penicillium citrinum and Candida albicans. In 2007, Yang Qiong et al. reported that the common pathogenic bacteria in external auditory canal fungal diseases are Aspergillus niger, Aspergillus fumigatus, Mucor, Trichophyton mentagrophytes, etc.

[0004] Enrofloxacin has the activity of transaminase and topoisomerase IV, interfering with the replication, transcription and repair of bacterial DNA, 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 class of glucocorticoids, which has anti-inflammatory and immunosuppressive effects. It is commonly used to treat a variety of inflammatory diseases, such as allergic diseases, rheumatic immune diseases, skin diseases, etc. Triamcinolone acetonide can relieve symptoms by reducing local inflammatory reactions and is commonly used for injection or topical application. Due to its immunosuppressive effect, side effects such as osteoporosis and gastrointestinal discomfort should be noted when used for a long time. The molecular structure is as follows:

[0007]

[0008] Gentamycin sulfate is an aminoglycoside antibiotic with a broad-spectrum antibacterial effect. It is mainly used to treat severe infections caused by sensitive bacteria, such as sepsis, pneumonia, peritonitis, etc. It kills bacteria by inhibiting bacterial protein synthesis. Because it may cause ototoxicity and nephrotoxicity, it needs to be used with caution. Blood drug concentration and renal function are usually monitored under the guidance of a doctor. The molecular structure is as follows:

[0009]

[0010] Pulegone is a monoterpene compound extracted from the mint plant, often used as a fragrance, fragrance and drug ingredient. It has a cooling effect and has certain analgesic, anti-inflammatory and antibacterial effects. Pulegone is often 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 local antipruritic. The 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 inflammation in pets. Summary of the invention

[0013] The main purpose of the present invention is to provide a veterinary enrofloxacin compound gel ear drops, specifically a compound gel ear drops prepared by combining four active substances: enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone. The compound gel ear drops have a significant therapeutic effect in treating inflammation, especially in treating inflammation of the external auditory canal.

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

[0015] The invention discloses a veterinary enrofloxacin compound gel ear drops, which is 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; 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, stir at 5-15° C., then add benzyl alcohol, stir until completely dissolved to obtain solution A, and set aside;

[0018] 2) dissolving rapeseed oil in 80-90% or more ethanol, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, adjusting the pH value to obtain solution B, and setting aside;

[0019] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 5-15°C, stir while adding, and keep stirring at 150 rpm / min during the adding process; after all the 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, stir at 5-15° C., then add benzyl alcohol, stir until completely dissolved to obtain solution A, and set aside;

[0023] 2) dissolving rapeseed oil in 80-90% or more ethanol, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, adjusting the pH value to 4.0-5.0 with citric acid to obtain solution B for later use;

[0024] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 5-15°C, stir while adding, and keep stirring at 150 rpm / min during the adding process; after all are 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] Further, 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 to poloxamer 184 is 5:0.7, and the ratio of carbomer 934P to 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 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 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, wherein poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3.

[0031] Further, 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, stir at 10°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside;

[0033] 2) dissolving rapeseed oil in ethanol of 85% or more, adding enrofloxacin, triamcinolone acetonide, pulegone and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 4.5, and setting aside;

[0034] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 10°C, and stir at a speed of 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 drugs for treating inflammation of the external auditory canal of animals. The animals can 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) In the compound gel of the present invention, enrofloxacin, triamcinolone acetonide, gentamicin sulfate and pulegone are mixed in a specific weight ratio, and can exert a synergistic effect in pharmacodynamic experiments, and the drug effect of the gel prepared by replacing or lacking one active substance is more significant; the compound gel of the present invention has a synergistic antibacterial effect and has a good therapeutic effect on external ear canal inflammation of animals.

[0039] 2) The compound gel ear drops of the present invention optimize the auxiliary material components and auxiliary material ratios to improve the stability of the compound gel. After accelerated experiments, it was found that the four active ingredients were not significantly reduced. In terms of improving drug permeability, the inventors made creative choices. The selected auxiliary materials such as benzyl alcohol and L-cysteine ​​can promote the transdermal penetration of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone, and exert better therapeutic effects. 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] Fig. 9 : The levels of pulegone at 0, 1, 2, 3, and 6 months. DETAILED DESCRIPTION

[0049] The present invention is further described below by way of examples. It should be correctly understood that the examples of the present invention are only some representative formulations, and their purpose is only to illustrate the present invention, rather than to limit the present invention. Therefore, simple improvements to the present invention under the premise of the method of the present invention all belong to the scope of protection claimed in 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, stir at 10°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside;

[0054] 2) dissolving rapeseed oil in ethanol of 85% or more, adding enrofloxacin, triamcinolone acetonide, pulegone and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 4.5, and setting aside;

[0055] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 10°C, and stir at a speed of 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, pulegone =: 1.2: 0.8: 0.3: 0.1; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, ethanol; 21: 1.1: 2.1: 0.3: 1.5: 45: 13; among which poloxamer 407 and poloxamer 184 are 3: 0.5, and carbomer 934P and carbomer 941 are 0.2: 5;

[0059] The preparation process is:

[0060] 1) Add poloxamer, L-cysteine ​​and carbomer to water, stir evenly, stir at 5°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside;

[0061] 2) dissolving rapeseed oil in ethanol of 80% or more, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 4.0, and setting aside;

[0062] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 5°C, and stir at a speed of 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, enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone are 3.6:1.2:0.9:0.4; poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol are 44:1.4:4.7:0.8:3.5:80:26; among them, poloxamer 407 and poloxamer 184 are 6:0.9, and carbomer 934P and carbomer 941 are 0.7:10.

[0066] The preparation process is:

[0067] 1) Add poloxamer, L-cysteine ​​and carbomer to water, stir evenly, stir at 15°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside;

[0068] 2) dissolving rapeseed oil in 90% or more ethanol, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 5.0, and setting aside;

[0069] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 15°C, and stir at a speed of 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, among 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 ethanol of 85% or more, adding L-cysteine, enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 4.5, and setting aside;

[0096] 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 10°C, and stir at a speed of 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, stir at 10°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside;

[0103] 2) dissolving rapeseed oil in ethanol of 85% or more, 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, control the temperature to 10°C, and stir at a speed of 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 hyphae suspension: Take an appropriate amount of Aspergillus fumigatus spores and inoculate them into 250 ml sterilized Sabouroud liquid culture medium. After culturing in a constant temperature shaking water bath at 37°C for 48 h, centrifuge at a radius of 15 cm, 2000 r / min, and centrifuge for 5 min. Discard the supernatant, transfer the precipitate to a 1500 μL EP tube, grind until the mycelium 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 selected 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 rat model of external otic fungal disease

[0112] All rats were anesthetized by abdominal injection of 10% chloral hydrate 3ml / Kg, and the external auditory canals of both ears of rats were selected as the research objects. Sterile sandpaper was rolled into a stick and gently scratched in the shape of "*" to avoid bleeding and damage to the tympanic membrane, thereby causing natural external auditory canal injury in clinical practice, 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; in the normal control group, 0.9% sodium chloride solution was applied to the corresponding scratched damaged parts 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 respective examples), with a thickness of about 0.1 mm, and used continuously for 7 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 Integral statistical analysis

[0120] Graphpadprism 9.0 software was used to analyze the data. P<0.05 was considered statistically significant.

[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); see the comparison between Example 1-3 and Comparative Example 1-2 for details. Figure 1 It can be seen that "**" Examples 1-3 are significantly different from Comparative Examples 1-4 (P<0.01).

[0123] 2. Transdermal Diffusion Experiment

[0124] 2.1. Drugs under investigation

[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 drugs were punched into round shapes (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, pulegone) was determined by high performance liquid chromatography, and the transdermal diffusion of the drugs in different embodiments was compared.

[0128] 2.3 Cumulative transmission detection

[0129] Results Figure 2-5 .

[0130] 2.4 Statistical analysis

[0131] Graphpadprism 9.0 software was used to analyze the data. P<0.05 was considered statistically significant.

[0132] 2.5 Results Analysis

[0133] Figure 2-5It can be seen from the skin penetration process that the compound gel of the present invention forms a highly synergistic system by carefully designing and optimizing the mutual compatibility of various excipients, which can effectively promote the penetration of active ingredients of drugs into the skin, thereby significantly improving their penetration. These excipients not only help to improve the solubility and stability of the drug, but also act synergistically on the skin barrier through multiple mechanisms to enhance the ability of drug molecules to pass through 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 in the deep layer of 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: Take triamcinolone acetonide, dissolve it in 70% methanol solution and dilute it to make a solution containing about 5 μg per 1 ml. Take 20 μL and inject it into the liquid chromatograph and 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 the solution, add 900 μL of 50% methanol, shake evenly, accurately measure 20 μL and inject into the liquid chromatograph, and 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 filler (pH adaptation range 0.8-8.0); 0.2 mol / L trifluoroacetic acid solution-methanol (96:4) as mobile phase; flow rate of 0.6-0.8 ml per minute; evaporative light scattering detector (high temperature non-divided flow mode: drift tube temperature of 105-110°C, carrier gas flow rate of 2.5 L per minute; low temperature diverted flow mode: drift tube temperature of 45-55°C, carrier gas pressure of 350 kPa) for determination.

[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 about 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 the solution, add 900 μL of 50% methanol, shake evenly, accurately measure 20 μL and inject into the liquid chromatograph, and 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: Chromatographic column: Thermos; Mobile phase: methanol: water = 60:40; Detection wavelength: 256nm

[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 mark with pure methanol, shake thoroughly, take 100 μL of solution, add 900 μL of 50% methanol, shake evenly, and inject for analysis;

[0150] Reference solution: Take 0.5g of enrofloxacin and dilute it to the corresponding scale with pure methanol, shake it thoroughly, take 100μL of the solution, add 900μL of 50% methanol, shake it evenly, and measure it in the same way. Calculate it by peak area using the external standard method to get the result.

[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 evenly, 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 238nm, flow rate is: 1ml / min, injection volume is 5μL.

[0156] 3.5 Testing conditions

[0157] Accelerated test: The compound gel prepared in the examples and comparative examples was placed as a test sample at a temperature of 40±2°C and a relative humidity of 75±5% for 6 months. The contents of the above 4 active substances were detected at 0, 1, 2, 3, and 6 months, respectively, and calculated by peak area using the external standard method.

[0158] 3.6 Test results

[0159] pass Figure 6-9 It can be seen that the contents of enrofloxacin, triamcinolone acetonide, gentamicin sulfate and pulegone in Example 1 are significantly higher than those in Comparative Examples 4-5 and Comparative Examples 8-9 after the accelerated test, indicating that the use of different excipients or additives, or different processes has a relatively large impact on the four effective substances. This reflects the rationality, superiority and creativity of the formula provided in this application, and the transformation and implementation of this application is expected to have excellent market prospects.

[0160] In summary, the compound gel formula provided in this application embodies a high degree of compatibility rationality, superiority and innovation. By accurately selecting and optimizing the ratio of various excipients, the transdermal absorption of the drug is significantly improved, while ensuring the stability and efficacy of the drug ingredients. This formula has shown significant advantages in enhancing drug permeability, prolonging the duration of drug efficacy and reducing side effects, and has good market prospects and commercial potential.

Claims

1. A veterinary enrofloxacin compound gel ear drops, characterized in that: The veterinary enrofloxacin compound gel ear drops are composed of the following components in parts by weight: the ratio of enrofloxacin, triamcinolone acetonide, gentamicin sulfate, and pulegone is 1.2-3.6: 0.8-1.2: 0.3-0.9: 0.1-0.4; the ratio of poloxamer, carbomer, benzyl alcohol, L-cysteine, low-erucic acid rapeseed oil, water, and ethanol is 21-44: 1.1-1.4: 2.1-4.7: 0.3-0.8: 1.5-3.5: 45-80: 13-26; The preparation of the veterinary compound gel ear drops comprises the following steps: 1) Add poloxamer, L-cysteine ​​and carbomer to water, stir evenly, stir at 5-15° C., then add benzyl alcohol, stir until completely dissolved to obtain solution A, and set aside; 2) dissolving rapeseed oil in 80-90% or more ethanol, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, adjusting the pH value to obtain solution B for later use; 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 5-15°C, stir while adding, and keep stirring at 150 rpm / min during the adding process; after all the solution is added, ensure uniform mixing to obtain solution C, which is set aside; 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

2. The veterinary enrofloxacin compound gel ear drops according to claim 1, characterized in that: 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.

3. The veterinary enrofloxacin compound gel ear drops according to claim 2, characterized in that: 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.

4. The veterinary enrofloxacin compound gel ear drops according to claim 3, characterized in that: 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, wherein the ratio of poloxamer 407 to poloxamer 184 is 5:0.7, and the ratio of carbomer 934P to carbomer 941 is 0.4:7.

3.

5. The veterinary enrofloxacin compound gel ear drops according to claim 1, characterized in that: The pH value is 4.0-5.0, and the pH adjuster is citric acid.

6. The veterinary enrofloxacin compound gel ear drops according to claim 1, characterized in that: The preparation of the veterinary compound gel ear drops comprises the following steps: 1) Add poloxamer, L-cysteine ​​and carbomer to water, stir evenly, stir at 5-15° C., then add benzyl alcohol, stir until completely dissolved to obtain solution A, and set aside; 2) dissolving rapeseed oil in 80-90% or more ethanol, adding enrofloxacin, triamcinolone acetonide, pulegone, and gentamicin sulfate, adjusting the pH value to 4.0-5.0 with citric acid to obtain solution B for later use; 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 5-15°C, stir while adding, and keep stirring at 150 rpm / min during the adding process; after all are added, ensure uniform mixing to obtain solution C, which is set aside.

7. The veterinary enrofloxacin compound gel ear drops according to claim 1, characterized in that: The veterinary enrofloxacin compound gel ear drops have the following components calculated by weight: 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, wherein poloxamer 407 and poloxamer 184 are 5:0.7, and carbomer 934P and carbomer 941 are 0.4:7.3; The veterinary enrofloxacin compound gel ear drops comprises the following steps: 1) Add poloxamer, L-cysteine ​​and carbomer to water, stir evenly, stir at 10°C, then add benzyl alcohol, stir until completely dissolved to obtain solution A, set aside; 2) dissolving rapeseed oil in ethanol of 85% or more, adding enrofloxacin, triamcinolone acetonide, pulegone and gentamicin sulfate to obtain solution B, adding citric acid to adjust the pH value to 4.5, and setting aside; 3) Slowly add solution B to the gel matrix of solution A, control the temperature to 10°C, and stir at a speed of 150 rpm / min; after all the solution is added, ensure uniform mixing to obtain solution C, which is set aside; 4) Filtering solution C through a 0.22 μm filter membrane to obtain a compound gel of enrofloxacin-triamcinolone acetonide-pulegone-gentamicin sulfate.

8. Use of the veterinary enrofloxacin compound gel ear drops according to claim 1 in preparing a drug for treating inflammation of the external auditory canal of animals.

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