Compound ivermectin juglone liniment as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of veterinary treatment medicines, and relates to a compound ivermectin juglone liniment as well as a preparation method and application thereof. The compound ivermectin juglone liniment is prepared from the following components in percentage by weight: 0.2 percent to 0.7 percent of juglone, 0.3 percent to 0.7 percent of ivermectin, 0.8 percent to 1 percent of tween-80, 0.1 percent to 0.15 percent of butylated hydroxytoluene, 0.2 percent to 5 percent of azone, 0.05 percent to 5 percent of menthol, 1 percent to 12 percent of isopropyl myristate and the balance of medium chain triglyceride, the compound ivermectin juglone liniment can be used for effectively removing parasites of animals, and is non-toxic and low in toxicity.

Description

Technical Field

[0001] The present invention belongs to the technical field of veterinary therapeutic drugs, and relates to a compound ivermectin juglone liniment and its preparation method and application. Background Art

[0002] The pathogens of rabbit mange are mainly mites of the families Sarcoptidae and Psoroptidae, which parasitize on the surface or in the epidermis of rabbits, thereby causing ectoparasitic diseases. Among them, adult Sarcoptes mites are 0.3-0.5 mm long, with a relatively small body size and are difficult to observe with the naked eye. They have 4 small legs, and their bodies are round or oval, with bristles and spines on the surface. They mainly parasitize in the host skin epidermis and feed on the host tissue fluid and interstitial fluid. Psoroptes mites are larger than Sarcoptes mites. Adult Psoroptes mites are 0.6-1.0 mm long, with an oblong body, long and thin legs and suckers, and bristles on the body surface. They mainly parasitize on the host skin surface and feed on the host lymph fluid and tissue fluid. Moreover, Psoroptes mites can also invade the rabbit ears, causing otitis externa, and even epilepsy in severe cases.

[0003] Ivermectin is a semi-synthetic macrolide antiparasitic drug produced by fermenting the soil microorganism Streptomyces avermitilis. It has the advantages of broad spectrum, high efficiency, small dosage and safety. Ivermectin has a good killing effect on internal and external parasites, especially nematodes and arthropods. Ivermectin can be used to prevent and treat various parasitic diseases of livestock and poultry. For example, it has a good killing effect on Otodectes cynotis and Sarcoptes scabiei in dogs, and can also prevent canine heartworm disease. Moreover, a single dose can kill internal and external parasites at the same time, so it has been widely used. After ivermectin reaches the skin surface or penetrates through the skin into the subcutaneous, dermal and adipose layers, it has a long retention time and can produce a lasting and good effect of killing ectoparasites.

[0004] Juglone, also known as 5-hydroxy-1,4-naphthoquinone, numerous studies at home and abroad have shown that it has good anti-tumor effects. In addition, it also has anti-inflammatory, antioxidant, antibacterial activity, hypoglycemic effects, etc. Juglone can be used as a raw material for dyes, pigments and spices, pH indicators and medical hemostatic agents, etc. It can also be used to treat eczema, herpes, constipation, impotence and spermatorrhea, low back pain, scrofula, and can also be used for the compound treatment of diabetes, gastritis, anemia, etc. In addition, juglone can also be used externally to treat diseases such as tinea capitis, psoriasis and elephantiasis. At present, it has been confirmed that it has a good acaricidal effect.

[0005] At present, the combined use of ivermectin and juglone has been disclosed. For example, a drug composition containing juglone and ivermectin for preventing and treating animal mange and its application disclosed in Chinese Patent 202310131771.3. This patent relates to a drug composition for preventing or treating animal mange, and its main components include juglone and ivermectin, and the mass ratio of the two is (1-10):(1-10). In order to dissolve juglone better, dimethyl sulfoxide needs to be used as a solvent.

[0006] The combined use of juglone and ivermectin has a synergistic effect, which can improve the control effect, reduce the losses caused by mites, and promote the recovery of damaged fur. However, although dimethyl sulfoxide (DMSO) has low acute toxicity, long-term exposure or high-dose use may cause adverse reactions in animals such as weight loss, hematological changes, liver and kidney damage. In addition, DMSO has permeability to the skin, which may cause skin irritation, itching, burning sensation, etc., and inhaling its vapor may also cause irritation to the respiratory tract. In the application of veterinary drugs, the use of pharmaceutical compositions containing dimethyl sulfoxide may pose certain health risks to livestock, especially when used for a long time or at high doses. Summary of the Invention

[0007] The purpose of the present invention is to avoid the deficiencies in the prior art and provide a compound ivermectin juglone liniment, its preparation method and application. The compound ivermectin juglone liniment can effectively remove parasites from animals and has low toxicity.

[0008] To achieve the above purpose, the present invention provides the following technical solutions:

[0009] Provide a compound ivermectin juglone liniment, which is composed of the following components in weight percentage:

[0010] Juglone 0.2% - 0.7%

[0011] Ivermectin 0.3% - 0.7%

[0012] Tween-80 0.8% - 1%

[0013] Butylated hydroxytoluene 0.1 - 0.15%

[0014] Azone 0.2% - 5%

[0015] Menthol 0.05% - 5%

[0016] Isopropyl myristate 1% - 12%

[0017] The balance is medium-chain triglyceride.

[0018] In some embodiments, the weight percentages of the components are as follows:

[0019] Juglone 0.5%

[0020] Ivermectin 0.5%

[0021] Tween-80 1%

[0022] Butylated hydroxytoluene 0.12%

[0023] Azone 0.5% - 5%

[0024] Menthol 0.1% - 5%

[0025] Isopropyl myristate 2% - 10%

[0026] The balance is medium-chain triglyceride.

[0027] Also provided is a preparation method of the above compound ivermectin juglone liniment, comprising the following steps:

[0028] Step 1: Mix the formula amounts of dibutylhydroxytoluene, juglone, ivermectin, and medium-chain triglyceride and stir evenly, and place them at 20°C - 30°C for ultrasonic dissolution to obtain a mixed solution;

[0029] Step 2: Add the formula amounts of triglyceride, azone, menthol, isopropyl myristate, and Tween-80 to the above mixed solution, and vortex and mix well to obtain the compound ivermectin juglone liniment.

[0030] In some embodiments, in Step 1, the temperature of ultrasonic dissolution is 25°C.

[0031] In some embodiments, the obtained compound ivermectin juglone liniment is stored at 40°C for at least 10 days. Also provided is the application of the above compound ivermectin juglone liniment, applying the compound ivermectin juglone liniment to the position of the animal skin surface infected with parasitic diseases.

[0032] In some embodiments, the animals include cattle, sheep, and rabbits.

[0033] In some embodiments, the parasites include sarcoptic mites and scab mites.

[0034] The beneficial effects of a compound ivermectin juglone liniment of the present invention:

[0035] The compound ivermectin juglone liniment of the present invention. Juglone and ivermectin in this compound liniment have good pesticidal effects on internal and external parasites in animals, especially nematodes and arthropods such as Otodectes cynotis and Sarcoptes scabiei in dogs. When used in combination, they can effectively improve the pesticidal effect on parasites. Moreover, the present invention overcomes the traditional method of using toxic solvents to dissolve the poorly soluble substance juglone, effectively ensuring the effect of the combined use of juglone and ivermectin. That is, the present application improves the combined use effect of juglone and ivermectin through the method of breathable penetration enhancement. Specifically: Tween-80, which can be used as a food and drug, is used as a non-ionic surfactant to achieve the functions of emulsifier, dispersant, solubilizer or stabilizer; dibutylhydroxytoluene is a food and drug grade chemical. As a phenolic antioxidant, it has the advantages of strong antioxidant property, high stability, low volatility and good durability; propylene glycol has the functions of solubilization and uniform mixing of each component, can penetrate into the stratum corneum and accumulate therein, increasing the solubility and partitioning of the drug in the stratum corneum, thereby promoting the transdermal penetration of the drug; azone has an obvious transdermal penetration enhancement effect on lipophilic, hydrophilic drugs and active ingredients and is a highly efficient transdermal penetration enhancer; menthol is a skin absorption promoter, which has the advantages of low price, wide source and small toxic and side effects; isopropyl myristate belongs to fatty acid ester transdermal absorption promoters and is the most widely used fatty acid ester in transdermal drug delivery systems; medium-chain triglycerides are the simplest fatty alcohols containing phenyl groups as non-toxic non-aqueous solvents. Therefore, all components of the present invention are environmentally friendly and non-toxic medicaments, and overcome the problem that juglone needs to use dimethyl sulfoxide to achieve synergistic use with ivermectin, effectively reducing the usage amount of toxic chemically synthesized drugs, and being more conducive to the combined action of compound ivermectin and juglone in treating parasites on animal skin. Brief Description of the Drawings

[0036] Figure 1 It is a comparison diagram of the mixed liquid of the ivermectin and juglone liniment prepared in Example 1 and Comparative Example 1 of the present invention. Detailed Description of the Embodiments

[0037] The preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the preferred embodiments of the present invention are shown in the drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to convey the scope of the present invention fully to those skilled in the art.

[0038] Example 1

[0039] This example discloses a compound ivermectin juglone liniment, with juglone being 0.2% - 0.7%, preferably 0.5%.

[0040] Ivermectin 0.3% - 0.7%, preferably 0.5%, Tween - 80 is preferably 1%, dibutylhydroxytoluene 0.1 - 0.15%, preferably 0.12%, azone 0.2% - 5%, preferably 0.5% - 5%, menthol 0.05% - 5%, preferably 0.1% - 5%,

[0041] Isopropyl myristate 1% - 12%, and the balance is medium - chain triglyceride.

[0042] Juglone and ivermectin in the above - mentioned compound ivermectin juglone liniment have good pesticidal effects on internal and external parasites in animals, especially nematodes and arthropods such as Otodectes cynotis and Sarcoptes scabiei in dogs. When the two are used in combination, the pesticidal effect on parasites can be effectively improved. Moreover, the present invention overcomes the traditional method of using toxic solvents to dissolve the poorly soluble substance juglone, effectively ensuring the effect of the combined use of juglone and ivermectin. That is, the present application improves the combined use effect of juglone and ivermectin through the way of breathable penetration enhancement. Specifically: Tween - 80, which can be used as a food and drug, is used as a non - ionic surfactant to realize the functions of emulsifier, dispersant, solubilizer or stabilizer; Dibutylhydroxytoluene is a food and drug - grade chemical. As a phenolic antioxidant, it has the advantages of strong antioxidant property, high stability, low volatility and good durability; Propylene glycol has the functions of solubilization and uniform mixing of each component, can penetrate into the stratum corneum and accumulate therein, increasing the solubility and partition of drugs in the stratum corneum, thus promoting the transdermal penetration of drugs; Azone has obvious transdermal penetration - enhancing effects on lipophilic, hydrophilic drugs and active ingredients and is a highly efficient transdermal penetration enhancer; Menthol is a skin absorption enhancer, which has the advantages of low price, wide source and small toxic and side effects; Isopropyl myristate belongs to fatty acid ester - type transdermal absorption enhancers and is the most widely used fatty acid ester in transdermal drug delivery systems; Medium - chain triglyceride is the simplest fatty alcohol containing phenyl as a non - toxic non - aqueous solvent. Therefore, all components of the present invention are environmentally friendly and non - toxic medicaments, and overcome the problem that juglone needs to use dimethyl sulfoxide to achieve synergistic use with ivermectin, effectively reducing the usage amount of toxic chemical synthetic drugs, and being more conducive to the combined action of compound ivermectin and juglone in treating parasites on the skin of animals.

[0043] The preparation method of the above - mentioned compound ivermectin juglone liniment includes the following steps:

[0044] Step 1: Mix the formula amounts of dibutylhydroxytoluene, juglone, ivermectin and medium - chain triglyceride and stir evenly. Place it at 20 - 30°C, preferably at 25°C, and obtain a mixed solution after ultrasonic dissolution. The color of the mixed solution is as Figure 1 shown.

[0045] Under ultrasonic-assisted pulverization at a certain temperature, the dissolution of juglone can be effectively improved, enhancing the medicinal effect and laying a foundation for avoiding the use of toxic solvents.

[0046] Step 2: Add the formulated amounts of triglyceride, azone, menthol, isopropyl myristate, and Tween-80 to the above-mentioned mixture, and vortex thoroughly to obtain the compound ivermectin juglone liniment.

[0047] In this example, the obtained compound ivermectin juglone liniment was stored at 40 °C for at least 10 days.

[0048] The presence of the compound ivermectin juglone liniment at a certain temperature can increase the drug content of the entire compound ivermectin juglone liniment and achieve efficient insecticidal effects.

[0049] Apply the compound ivermectin juglone liniment to the positions on the animal skin surface infected with parasites.

[0050] Among them, the animals include cattle, sheep, and rabbits.

[0051] The parasites include sarcoptic mites and scab mites.

[0052] Comparative Example 1

[0053] To illustrate that the way of mixing ivermectin and juglone in the corresponding solvent in the present invention is different from the traditional way of mixing ivermectin and juglone using dimethyl sulfoxide, the following comparative experiment was carried out:

[0054] Dissolve ivermectin and juglone in a dimethyl sulfoxide solution to obtain Figure 1 the shown color.

[0055] Among them, since the color of dimethyl sulfoxide itself is colorless, from Figure 1 comparison, it can be seen that the color of the solution A after dissolving ivermectin and juglone in dimethyl sulfoxide becomes black, and the mixture B obtained by mixing ivermectin and juglone in Example 1 is a light yellow mixture. This shows that the method of treating ivermectin and juglone in the present invention is not the same as the traditional method of dissolving ivermectin and juglone using dimethyl sulfoxide. The present invention improves the synergistic effect of ivermectin and juglone through the transdermal penetration method.

[0056] Experimental Example

[0057] To further illustrate the test process of the present invention and the resulting effects, the following experiment was carried out.

[0058] Experimental Example 1 In vitro transdermal test

[0059] This experiment will specifically demonstrate the in vitro transdermal test method used in the single-factor test and orthogonal test for screening the prescription.

[0060] Before the in vitro transdermal test, healthy Kunming mice were killed by cervical dislocation, and the abdominal hair was removed with a depilatory knife. The abdominal skin was peeled off, and the remaining hair was removed with a depilatory cream, and the mouse skin was washed with saline. The muscles and blood vessels and nerves on the inside of the skin were wiped with an absorbent cotton ball, and the fat in the skin was removed. The treated mouse skin was stored in a -20℃ refrigerator and used within 24 hours. It was thawed before use.

[0061] The experimental method used for screening the prescription of the compound ivermectin juglone ointment of the present invention is an in vitro transdermal test, and the device used is an in vitro transdermal diffusion device. The transdermal diffusion test simulates the skin system through an in vitro diffusion cell device. The experiment uses a modified Franz diffusion cell device to fix the treated in vitro mouse back skin between the sample cell and the receiving cell, with the stratum corneum facing the sample cell and the dermis facing the receiving cell. During the experiment, the homemade juglone ointment was added to the sample cell, and the sample cell and the receiving cell were sealed with a sealing film. The magnetic stirrer speed was set to 500r / min and the temperature was 35°C. 1mL was sampled at 12h and 24h respectively. After each sampling, an equal volume of receiving liquid at the same temperature was added to the receiving cell. The sampled liquid was filtered through a 0.22μm microporous filter membrane, and the sample was tested according to the above detection method. The content of juglone and ivermectin was calculated, and the cumulative permeation per unit area (Qn, μg / cm 2 ) is calculated as

[0062]

[0063] Where Cn is the drug concentration in the receiving solution at each sampling point (μg / mL), V is the volume of the receiving solution, Ci is the drug concentration in the receiving solution at time i (i≤n-1) (μg / mL), Vi is the sampling volume, and A is the effective diffusion area (cm 2 ).

[0064] Experimental Example 2 Screening of Receptor Fluid

[0065] This experimental example will specifically demonstrate the specific steps of screening the in vitro transdermal receiving solution before performing single factor screening.

[0066] The experiment investigated the saturated solubility of juglone and ivermectin in eight receiving solutions: normal saline, PBS, 10% ethanol-normal saline, 10% ethanol-PBS, 30% ethanol-normal saline, 30% ethanol-PBS, 60% ethanol-normal saline, and 60% ethanol-PBS.

[0067] Excess juglone and ivermectin bulk drugs were separately taken and added into 15 mL centrifuge tubes. 5 mL of normal saline, PBS, 10% ethanol - normal saline, 10% ethanol - PBS, 30% ethanol - normal saline, 30% ethanol - PBS, 60% ethanol - normal saline, and 60% ethanol - PBS were separately added into different centrifuge tubes, with 3 parallels set for each group. After adding the drugs and the receiving solutions, the centrifuge tubes were vortexed thoroughly and then ultrasonicated for 10 min to fully mix and dissolve the drugs and the receiving solutions. After observing that no more drugs dissolved, the centrifuge tubes were placed on an oscillator and shaken thoroughly at 25 °C and 200 r·min -1 under the condition for 24 h. After shaking, each receiving solution was taken out to observe whether there was precipitation. If precipitation existed, it was taken out and centrifuged at 10000 r·min -1 for 10 min. The supernatant was aspirated, diluted with the mobile phase, filtered through a membrane, and then analyzed by HPLC to screen out the receiving solutions with relatively high saturated solubility of the drugs for subsequent experiments.

[0068] As can be seen from Table 1, the saturated solubilities of the two drugs were relatively high in 30% ethanol - normal saline, 30% ethanol - PBS, 60% ethanol - normal saline, and 60% ethanol - PBS. Therefore, these four receiving solutions were selected for in vitro transdermal experiments, and the receiving solutions with the highest cumulative permeation amounts at 12 h and 24 h were screened out for subsequent experiments. As can be seen from Table 2, when the receiving solution was 60% ethanol - normal saline, the cumulative permeation amounts of the two drugs at 12 h and 24 h were the highest. Therefore, 60% ethanol - normal saline was selected as the receiving solution.

[0069] Table 1 Saturated Solubilities of Drugs in Different Receiving Solutions

[0070]

[0071] Table 2 24H Cumulative Permeation Amounts of Drugs in Different Receiving Solutions (n = 3)

[0072]

[0073]

[0074] Experimental Example 3 Preparation Method of Compound Ivermectin Juglone Liniment

[0075] This experimental example will specifically demonstrate the preparation method of compound ivermectin juglone liniment. The specific experiments and preparation methods are as follows:

[0076] Juglone is insoluble in cold water but slightly soluble in hot water, and can be dissolved in ethanol, ether, and miscible with chloroform and benzene. These information indicate that juglone has poor solubility in water, but has a certain solubility in hot water; ivermectin is almost insoluble in water, soluble in methanol, ethyl acetate or chloroform, and soluble in ethanol or acetone. In order to enable the drug to better penetrate into the animal body to exert its efficacy for better therapeutic effects, the penetration enhancers azone, menthol, and isopropyl myristate are selected as penetration enhancers, the solvent is medium-chain triglyceride, the preservative is dibutylhydroxytoluene, and the solubilizer is Tween-80.

[0077] Excess amounts of juglone and ivermectin raw materials were respectively taken and added to 10 mL centrifuge tubes. 2 mL of medium-chain triglyceride, isopropyl myristate, azone, and Tween-80 were respectively added. After sufficient vortexing, ultrasonic treatment was carried out for 10 min to fully mix and dissolve the drug with the receiving solution. After observing that no more drug continued to dissolve, the centrifuge tubes were placed on an oscillator and shaken thoroughly at 25 °C and 200 r·min-1 for 24 h. After shaking, observe whether there is precipitation. If there is precipitation, centrifuge at 10000 r·min-1 for 10 min after taking it out, aspirate the supernatant, dilute it with the mobile phase and filter through a membrane and then load it onto the machine.

[0078] The test results show that the saturated solubility of juglone in medium-chain triglyceride is 10.16 mg / mL, in isopropyl myristate is 7.64 mg / mL, in azone is 0.33 mg / mL, and in Tween-80 is 3.90 mg / mL; the saturated solubility of ivermectin in medium-chain triglyceride is 20.58 mg / mL, in isopropyl myristate is 8.30 mg / mL, in azone is 179.14 mg / mL, and in Tween-80 is 62.63 mg / mL.

[0079] The solubility of juglone and ivermectin in each excipient can provide a reference basis for the addition sequence of each excipient in the preparation process of the preparation. The preparation process of the compound liniment is as follows:

[0080] Step S1: Weigh the following components according to the percentage content, juglone 0.5%, ivermectin 0.5%, dibutylhydroxytoluene 0.12%, azone 0.5%-5%, menthol 0.1%-5%, isopropyl myristate 2%-10%, Tween-80 1%-2%, medium-chain triglyceride 50%-70%;

[0081] Step S2: Stir the dibutylhydroxytoluene, juglone, ivermectin, and medium-chain triglyceride weighed in Step S1 evenly according to the above percentages, and place them at 25 °C for ultrasonic dissolution to obtain a mixed solution;

[0082] Step S3: Add the medium-chain triglycerides, azone, menthol, isopropyl myristate, and Tween-80 weighed in Step S1 into the mixed solution obtained in Step S2, and vortex thoroughly to obtain the compound liniment.

[0083] Experimental Example 4 Single-factor Test of Compound Ivermectin Juglone Liniment

[0084] In this experimental example, the in vitro transdermal test will be used to screen out the optimal dosages of each excipient.

[0085] Using the in vitro transdermal receiving solution screened in Experimental Example 2 above, the dosages of each excipient screened are screened with the cumulative drug permeation amount in 24 h as the index. Five concentrations are set for each of the three penetration-enhancing components, azone, menthol, and isopropyl myristate, and three optimized concentrations with better penetration-enhancing effects are screened from each component through experiments. The obtained optimized concentrations are used for the orthogonal test to screen the best formulation. The five concentrations of azone are set as a1, a2, a3, a4, and a5 respectively, the five concentrations of menthol are b1, b2, b3, b4, and b5 respectively, and the five concentrations of isopropyl myristate are c1, c2, c3, c4, and c5 respectively.

[0086] (1) Investigation of the single factor of azone

[0087] Fix the dosages of menthol and isopropyl myristate as b3 and c3 respectively, set the dosages of azone as a1, a2, a3, a4, and a5 respectively, prepare liniment samples according to the liniment preparation method in Experimental Example 3 above, use the in vitro transdermal receiving solution screened in Experimental Example 2 above, sample the samples at 12 h and 24 h respectively, with a total of 5 groups of experiments, 3 parallels in each group, and a total of 15 groups of experiments. Filter the obtained samples through a 0.22 μm filter membrane, and detect the contents of juglone and ivermectin respectively.

[0088] It can be seen from Table 3 that when the azone concentration is a2, a4, and a5, the cumulative permeation amounts of ivermectin and juglone at 12 h and 24 h are relatively higher than those of other groups. Therefore, the three concentrations of a2, a4, and a5 are selected as the relatively optimal concentrations of azone for the orthogonal test.

[0089] Table 3 Results of the single-factor screening of azone

[0090]

[0091]

[0092] (2) Investigation of the single factor of menthol

[0093] The dosages of azone and isopropyl myristate are a3 and c3 respectively. The dosages of menthol are set as b1, b2, b3, b4, and b5. According to the preparation method of the liniment in Experimental Example 3 above, liniment samples are prepared respectively. Using the in vitro percutaneous absorption receptor solution screened in Experimental Example 2 above, samples are sampled at 12 h and 24 h respectively. There are a total of 5 groups of experiments, with 3 parallels in each group, for a total of 15 groups of experiments. The collected samples are filtered through a 0.22 μm filter membrane, and the contents of juglone and ivermectin are detected respectively according to the above sample detection method.

[0094] As can be seen from Table 4, when the menthol concentration is b1, b3, or b4, the cumulative permeation amounts of ivermectin and juglone are relatively higher than those of other groups at 24 h. At 12 h, the effects of menthol at concentrations b1, b2, b3, and b4 on the cumulative permeation amounts of ivermectin and juglone are not significantly different. Considering comprehensively, menthol at concentrations b1, b3, and b4 is selected as the preferred concentration for orthogonal experiment to screen the formulation.

[0095] Table 4 Results of single-factor screening of menthol

[0096]

[0097]

[0098] (3) Investigation of single factor of isopropyl myristate

[0099] The dosages of azone and menthol are a3 and b3 respectively. The dosages of isopropyl myristate are set as c1, c2, c3, c4, and c5. According to the preparation method of the liniment in Experimental Example 3 above, liniment samples are prepared respectively. Using the in vitro percutaneous absorption receptor solution screened in Experimental Example 2 above, samples are sampled at 12 h and 24 h respectively. There are a total of 5 groups of experiments, with 3 parallels in each group, for a total of 15 groups of experiments. The collected samples are filtered through a 0.22 μm filter membrane, and the contents of juglone and ivermectin are detected respectively according to the above sample detection method.

[0100] As can be seen from Table 5, when the concentration of isopropyl myristate is c1, c4, or c5, the cumulative permeation amounts of ivermectin and juglone are relatively higher than those of other groups at 12 h and 24 h. Therefore, three concentrations of c1, c4, and c5 are selected as the preferred concentrations of isopropyl myristate for orthogonal experiment.

[0101] Table 5 Results of single-factor screening of isopropyl myristate

[0102]

[0103] Experimental Example 5 Orthogonal experiment of compound ivermectin juglone liniment

[0104] This example will conduct an orthogonal experiment based on the single-factor experiment results of Example 4 to obtain the optimal formulation.

[0105] From the single-factor screening results of the above 4 penetration enhancers, when the azone concentrations are a2, a4, and a5, the cumulative permeation amounts of ivermectin and juglone at 12 h and 24 h are relatively higher than those of other groups. Therefore, the three concentrations of a2, a4, and a5 are selected as the relatively optimal concentrations of azone for the orthogonal experiment; when the menthol concentrations are b1, b3, and b4, the cumulative permeation amounts of ivermectin and juglone at 24 h are relatively higher than those of other groups. At 12 h, the menthol at the concentrations of b1, b2, b3, and b4 has little difference in the cumulative permeation amount of ivermectin and juglone. Considering comprehensively, the menthol at the concentrations of b1, b3, and b4 is selected as the relatively optimal concentration for the orthogonal experiment to screen the formulation; when the isopropyl myristate concentrations are c1, c4, and c5, the cumulative permeation amounts of ivermectin and juglone at 12 h and 24 h are relatively higher than those of other groups. Therefore, the three concentrations of c1, c4, and c5 are selected as the relatively optimal concentrations of isopropyl myristate for the orthogonal experiment.

[0106] Using the above in vitro transdermal experiment, the relatively optimal dosages of the selected excipients are screened with the 24-h cumulative drug permeation amount as the index. The three optimal dosages of each excipient screened by the single-factor experiment are selected for the orthogonal experiment. The orthogonal experiment uses the L25(3 4 ) experimental table, with 4 factors and 3 levels. Among them, 1, 2, and 3 represent the 3 levels of each factor respectively, and there are a total of 9 groups of experiments. The orthogonal experiment table is shown in Table 6 below, and the orthogonal experiment results are shown in Table 7 below.

[0107] Table 6 Orthogonal Experiment Table

[0108]

[0109] Table 7 Orthogonal Experiment Results

[0110]

[0111]

[0112] It can be seen from the orthogonal experiment table that the cumulative permeation amounts of ivermectin and juglone at 12 h and 24 h in the 3rd group of experiments are both relatively high. Therefore, the 3rd group of formulation is selected as the optimal formulation of the compound liniment, that is, the optimal formulation of the liniment is azone at a5, menthol at b4, and isopropyl myristate at c5.

[0113] Experimental Example 6 Influence Factor Test of Compound Juglone and Ivermectin

[0114] This experimental example uses the compound ivermectin juglone liniment prepared with the formulation components of the above experimental example 1 for experiments. According to the guiding principles for the stability test of raw drugs and preparations, this experiment will conduct stress tests on the prepared compound liniment to investigate the stability of the drug preparation.

[0115] (1) High-intensity light irradiation test

[0116] After determining the content of the best-formulated preparation sample screened out on the day of preparation, take an appropriate amount and place it in an open 50 mL glass bottle. Place it under a high-intensity light condition of (4500 ± 500) LX for 10 days. The total illuminance of the light source is not less than 1.2 * 10 6 lx·h, and sample on the 0th, 5th, and 10th days. Detect the content by HPLC, and at the same time observe the color change of the preparation. The test results are shown in Table 8 below.

[0117] Table 8 Results of content determination in the light irradiation experiment

[0118]

[0119] (2) High-temperature test

[0120] After determining the content of the best-formulated preparation sample screened out on the day of preparation, take an appropriate amount and place it in an open 50 mL glass bottle. Place it at 60 °C for 10 days, and sample on the 0th, 5th, and 10th days. Detect the content by HPLC, and at the same time observe the color change of the preparation. If the content of the test sample is lower than the specified limit, conduct the test in the same manner at 40 °C. It was found that it was unstable at 60 °C, and a high-temperature test was carried out at 40 °C. The test results are shown in Table 9 below.

[0121] Table 9 Results of content determination in the high-temperature test

[0122]

[0123] (3) High-humidity test

[0124] After determining the content of the best-formulated preparation sample screened out on the day of preparation, take an appropriate amount and place it in an open 50 mL glass bottle. Place it at 25 °C under conditions of relative humidity of 90% ± 5% for 10 days, and sample on the 0th, 5th, and 10th days. Detect the content by HPLC, and at the same time observe the color change of the preparation and accurately weigh the weight of the test sample before and after the experiment to investigate the hygroscopic and desorption properties of the test sample. If the moisture absorption weight gain is more than 5%, conduct the test in the same manner under conditions of relative humidity of 75% ± 5%. If the moisture absorption weight gain is less than 5% and other inspection items meet the requirements, this test will no longer be carried out.

[0125] It was found in the experiment that the content of juglone decreased significantly and was unstable under the condition of 95%±5% humidity. Therefore, the test sample was placed in an appropriate clean container with the opening exposed under the condition of 75%±5% humidity for 10 days. Samples were taken on the 5th and 10th days, and monitored according to the key stability investigation items. The results are shown in Table 10 below, indicating that the compound preparation has good stability under the condition of 75%±5% humidity.

[0126] Table 10 Results of content determination in high humidity experiment

[0127]

[0128] Experimental Example 7 Skin irritation test of compound juglone ivermectin liniment

[0129] In this experiment, the preparations prepared from the formula components of the above Experimental Example 1 (compound preparation), Experimental Example 2 (juglone single preparation) and Experimental Example 3 (blank excipient) were subjected to skin irritation tests, and the specific tests were divided into single-dose skin irritation test and multiple-dose skin irritation test.

[0130] (1) Single-dose skin irritation test

[0131] Twelve healthy New Zealand rabbits, half male and half female, about 3 months old, were purchased from Guangdong Experimental Animal Center, with a body weight of 1.7 - 2.2 kg, and divided into two groups, Group A and Group B, namely the intact skin group and the damaged skin group. After shaving the hair on the back of the animals, it was divided into four drug administration areas, a, b, c, and d. Among them, the bilateral drug administration sites of Group B were scratched with a sterile surgical blade in a "well" shape until epidermal bleeding occurred, with the degree of damage to the stratum corneum and epidermal bleeding, and disinfected with iodophor to avoid infection, as the damaged skin group.

[0132] On the day of the experiment, the rabbits were treated according to the four areas on the back, that is, a was not treated, b was given the blank excipient, c was given the juglone single liniment, and d was given the compound liniment. The administration dose was 0.2 mL. After administration, it was covered with a dressing for at least 4 h. After administration once, the residual test drug was removed with warm water. The application sites of the drug were observed for erythema and edema at 1, 24, 48, and 72 h after removing the test drug.

[0133] Referring to the experimental design and scoring criteria in the "Technical Guidelines for the Study of Irritation, Hypersensitivity and Hemolysis of Chemical Drugs", for the single-dose skin irritation test, the average value of the skin reaction scores at each time point of each group was calculated, and the irritation intensity was evaluated according to Table 11 below. The skin irritation intensity evaluation criteria are shown in Table 12.

[0134] Table 11 Skin irritation reaction scoring criteria

[0135]

[0136]

[0137] Table 12 Skin irritation intensity evaluation criteria

[0138]

[0139] The test results are shown in Tables 13 and 14. It can be seen from the tables that within 72 hours, the skin irritation scores of each area in the intact skin group were between 0 and 0.49, and the intensity was non-irritating; in the damaged skin group, individual test rabbits showed transient mild erythema in 4 areas 1 hour after removing the drug, and the skin irritation scores were between 0 and 2.99, with mild irritation, but it basically disappeared after 24 hours, and the skin irritation scores were between 0 and 0.49, and the skin irritation intensity was non-irritating. In summary, compound ivermectin juglone liniment and juglone single liniment have no single-dose skin irritation to the skin of test rabbits.

[0140] Table 13 Results of single-dose skin irritation test of compound liniment (n = 6)

[0141]

[0142] Table 14 Evaluation of the intensity of single-dose skin irritation test of compound liniment

[0143]

[0144] (2) Repeated-dose skin irritation test

[0145] Twelve New Zealand rabbits, half male and half female, about 3 months old, were purchased from Guangdong Laboratory Animal Center, with a body weight of 1.6 - 2.3 kg. They were divided into two groups, A and B, namely the intact skin group and the damaged skin group. After shaving the hair on the back of the animals, there were four drug administration areas, a, b, c, and d. Among them, in group B, the bilateral drug administration sites were scratched with a sterile surgical blade in a cross shape until epidermal bleeding occurred, with the damage limited to the stratum corneum and epidermal bleeding, and then disinfected with iodophor to avoid infection, serving as the damaged skin group.

[0146] On the day of the experiment, according to the four areas on the back of the rabbits, the following treatments were given: a was not treated, b was given blank excipients, c was given juglone single liniment, and d was given compound liniment. The administration dose was 0.2 mL. After administration, it was covered with a dressing for at least 4 hours. During the test period, the drugs were administered on the 1st and 3rd days. After the last administration, the residual test drugs were removed with warm water. The erythema and edema conditions of the drug-applied sites were observed 1, 24, 48, and 72 hours after removing the test drugs, and the irritation intensity was evaluated according to Tables 11 and 12.

[0147] The test results are shown in Tables 15 and 16. As can be seen from the tables, within 72 hours, the skin irritation scores of each area in the intact skin group were between 0 and 0.49, and the intensity was non-irritating; in the damaged skin group, individual test rabbits showed transient mild erythema in 4 areas 1 hour after removing the drug, and the skin irritation scores were between 0 and 2.99, belonging to mild irritation. However, it gradually disappeared after 24 hours, and the skin irritation scores were between 0 and 0.49, and the skin irritation intensity was non-irritating. In summary, the compound ivermectin juglone liniment and the juglone single liniment had no skin irritation after multiple administrations to the test rabbits.

[0148] Table 15 Results of multiple skin irritation tests of compound liniment (n = 6)

[0149]

[0150] Table 16 Evaluation of the intensity of multiple skin irritation tests of compound liniment

[0151]

[0152] Experimental Example 8 Pharmacodynamic test of compound liniment

[0153] (1) In vitro acaricidal test

[0154] In this experimental example, the liniments prepared from the formula components of the above-mentioned Example 1 (compound preparation), Example 2 (juglone single preparation), Example 3 (blank excipient), Example 4 (ivermectin single preparation), the commercially available ivermectin transdermal solution, and medium-chain triglycerides were used for the experiment. The sarcoptes scabiei used in the test were derived from rabbits naturally infected with sarcoptes scabiei. The diseased rabbits were purchased from a farm in Nanning, 3 months old, weighing about 2 kg, and were raised in a cool and ventilated room. The water and food were changed daily, and the bottom tray was cleaned and disinfected. The test was divided into 6 groups, namely the medium-chain triglyceride group, the blank excipient control group, the positive drug control group, the self-made juglone preparation group, the self-made ivermectin preparation group, and the self-made compound preparation group. Before each test, an appropriate amount of scab was taken from the ear of the sarcoptes scabiei-infected rabbit, and 10 scabies mites of similar size and good vitality were picked out with a mite-picking needle and placed in a 35 mm cell culture dish for in vitro acaricidal test.

[0155] The experiment was divided into 6 groups, namely the medium-chain triglyceride group, the blank excipient control group, the positive drug control group, the self-made juglone preparation group, the self-made ivermectin preparation group, and the self-made compound preparation group. Before the experiment, an appropriate amount of scab was taken from the ears of rabbits with sarcoptic mange, and 10 rabbit sarcoptes with similar sizes and better vitality were picked out with a mite-picking needle and placed in a 35 mm cell culture dish for the experiment. During the experiment, 10 mites were placed in each cell culture dish, and 100 μL of the drug solution was added to each dish. During the experiment, the 10 sarcoptes were all immersed in the drug solution. Each test group and control group were repeated 6 times. After adding the drug solution to the cell culture dish, the number of dead mites was observed and recorded every 3 min under a stereomicroscope until all the mites in the drug group died. The experiment was observed and recorded for 45 min in total. The following are the photos of the 6 experimental groups during the experiment. The criteria for determining the death of the insect body: Under the stereomicroscope, use a mite-picking needle to touch the limbs of the motionless mite. After touching, observe for 2 min. If the insect body does not move and its intestine does not show peristalsis, it can be determined as dead.

[0156] During the experiment, the number of dead mites in each group at each time point was observed and recorded every 3 min under a stereomicroscope. The experimental data were analyzed using IBM SPSS Statistics 26.0 software, and the median lethal time (LT50) of each drug group was calculated using Probit regression analysis. The in vitro acaricidal effects of each drug group are shown in Table 17 below. The time for the compound group to cause 100% death of sarcoptes was 9 - 12 min, the time for the single juglone group to cause 100% death of sarcoptes was 12 - 15 min, the time for the single ivermectin group to cause 100% death of sarcoptes was 21 - 24 min, the time for the commercially available drug group to cause the death of sarcoptes was 24 - 27 min, the blank excipient group had a certain acaricidal effect on sarcoptes during the experiment, and medium-chain triglycerides had no acaricidal effect on sarcoptes during the experiment.

[0157] Table 17 In vitro acaricidal effects (n = 6)

[0158]

[0159] The experimental data were analyzed using IBM SPSS Statistics 26.0 software, and the median lethal time (Medianlethal time, LT 50 ) was calculated using Probit regression analysis. The results are shown in Table 18 below. Based on the analysis of the median lethal time of the drug on sarcoptes, it was found that the LT 50 of the compound group, the single juglone group, and the single ivermectin group were 4.716 min, 6.750 min, and 12.837 min respectively. The commercially available drug group needed 36 - 39 min to achieve 100% mortality of sarcoptes, and its LT 50It is 24.157 min.

[0160] Table 18 Median lethal time (LT 50 )

[0161]

[0162] From the above data, it can be seen that the in vitro acaricidal effect of the compound preparation group is better than that of the juglone single - agent group and the ivermectin single - agent group, and the acaricidal effect of the combination of juglone and ivermectin is better than that of their individual use.

[0163] (2) In - vivo acaricidal test on rabbits

[0164] In this experiment, compound ivermectin juglone liniment, juglone single - agent liniment, and commercially available ivermectin transdermal solution were used for the test. The animals used in the test were 40 naturally ear - mite - infected rabbits (purchased from a rabbit farm in Shandong Province), including New Zealand rabbits, Belgian rabbits, and Himalayan rabbits, with weights ranging from 1.88 - 4.55 kg. During the test period, the experimental animals were raised in a cool and ventilated indoor environment, with water and food changed daily, and the bottom trays were cleaned and disinfected. After the experiment, all unhealed rabbits were treated.

[0165] The in - vivo experiment was divided into 4 groups, namely the infected non - medicated group (A), the self - made compound liniment group (B), the self - made juglone single - agent group (C), and the commercially available ivermectin transdermal solution group (D), with 10 diseased rabbits in each group, a total of 40 diseased rabbits. The groups were raised separately, and the diseased rabbits were raised in a ventilated rabbit house. The plastic pads on the bottom of the rabbit cages were changed every day, and the replaced plastic pads were soaked in a 2% ronnel aqueous solution for washing, and then dried after washing.

[0166] Before the test, the weight of each animal was weighed and recorded, and the initial diseased area of the left and right ears of each animal was estimated. The dosage was calculated based on the diseased area of the animal, and the dosage was 0.1 mL / cm 2 / kg. Among them, 1 - 40 are the individual numbers of the animals, L and R represent the left and right ears of the experimental animals respectively. The actual dosage of the left and right ears of each animal was calculated according to the weight of each animal, and the statistical results are shown in Table 19 below.

[0167] Table 19 Dosage statistics table

[0168]

[0169]

[0170] Continued Table 19 Dosage statistics table

[0171]

[0172]

[0173] Continued Table 19 Statistical Table of Drug Dosage

[0174]

[0175] During the experimental treatment period, the diseased rabbits were randomly divided into 4 groups. Group A was the infected group without drug administration, given normal saline; Group B was the group with the self-made compound liniment; Group C was the group with the self-made juglone single agent; Group D was the control group with the commercially available ivermectin transdermal solution. After the animals were randomly grouped, the body weight of each animal and the diseased area of the left and right ears were statistically recorded. The drug was administered at a dose of 0.1 mL / cm 2 / kg on the 1st day and the 3rd day respectively, and the ear canal scabs were scraped on the 2nd day, 4th day, 7th day, and 14th day respectively for microscopic examination under a stereomicroscope. During the test period, the following efficacy judgments were made:

[0176] a. Parasite negative conversion rate: During the test period, the drug was administered on the 1st day and the 3rd day respectively. On the 2nd day, 4th day, 7th day, and 14th day, 0.1 g of ear canal scabs were scraped, and the number of live mites and total mites in each gram of scab before and after drug administration were recorded. Calculate the parasite reduction rate and live mite rate. Among them, the parasite reduction rate = (number of live mites before drug administration - number of live mites after drug administration) / number of live mites before drug administration × 100%, the live mite rate = number of live mites / (number of live mites + number of dead mites) × 100%, and the parasite negative conversion rate (%) = number of animals with negative parasite conversion after drug administration / number of animals with positive parasites before drug administration × 100%. Among the experimental animals, those with live parasites detected were judged as positive, while those without live parasites detected were judged as negative.

[0177] b. Lesion score: After drug administration, the itching condition of the diseased rabbits and the recovery degree of the affected skin were observed and recorded every day as the efficacy judgment index. The scoring criteria are shown in Table 20 below.

[0178] Table 20 Grading System for Clinical Cases of Ear Mites Infection in Rabbits and Recovery Status

[0179]

[0180]

[0181] c. Body weight gain: Each group of rabbits was weighed on an empty stomach on the day before the test and after the test ended, and the weight changes before and after drug administration were statistically recorded. (Body weight gain = body weight after the test - body weight before the test.)

[0182] The test results are as follows:

[0183] a. Parasite negative conversion rate

[0184] During the experiment, 0.1 g of the scab in the ear canal of the diseased rabbits was scraped on the 2nd day, 4th day, 7th day, and 14th day respectively to calculate the reduction rate and negative conversion rate of the mites per gram of scab before and after medication. Among them, the reduction rate of mites = (the number of live mites before medication - the number of live mites after medication) / the number of live mites before medication × 100%, the live mite rate = the number of live mites / (the number of live mites + the number of dead mites) × 100%, and the negative conversion rate of mites (%) = the number of mites turning negative after medication / the number of positive mites before medication × 100%. Among the experimental individuals, the experimental animals with live mites detected were judged as positive, while those without live mites detected were judged as negative. The reduction rate, live mite rate, negative conversion rate, etc. of each drug group for each administration site of the experimental animals and each diseased animal were calculated respectively, and the therapeutic effects of each group of drugs on the diseased animals were statistically analyzed. The record form is shown in Tables 21 - 22. Among them, Group A is the non - medicated group, Group B is the compound juglone ivermectin liniment group, Group C is the single - ingredient juglone group, and Group D is the commercially available drug group.

[0185] Table 21 Therapeutic effects on the left and right ears of diseased rabbits

[0186]

[0187]

[0188] As can be seen from the above table, in the compound juglone ivermectin liniment group, there were no live mites in the right ear (B - R) of the experimental rabbits on the 2nd day, and the reduction rate of mites reached 100%. There were still a small number of live mites in the left ear, and the live mite rate was 0.91%. After the second administration, there were no live mites detected by microscopy in the compound juglone liniment group on the 4th day, and there was no recurrence within 14 days. On the 2nd day, the reduction rates of mites in the left and right ears of the single - ingredient juglone drug group were 97.4% and 99.3% respectively, both higher than those of the commercially available drug group and lower than 98.9% and 100% of the compound liniment group. After the second administration, there were no live mites detected by microscopy in the single - ingredient juglone liniment group on the 4th day, and there was no recurrence within 14 days. In the commercially available drug group, the reduction rates of mites in the left and right ears reached 98.9% and 97.2% respectively after the first administration, and the live mite rates were 1.6% and 12.3% respectively. After the second administration, the reduction rates of mites in the left and right ears of the animals were both 99.7%, and the live mite rates were 0.4% and 0.6% respectively. However, on the 7th day and 14th day, the live mite rates in the commercially available group increased.

[0189] Table 22 Therapeutic effects on the body of diseased rabbits

[0190]

[0191] Note: Different lowercase letters in the same column indicate significant differences (0.01 < p < 0.05), and different capital letters indicate extremely significant differences (p < 0.01)

[0192] The obtained data were summarized and processed using one-way analysis of variance and nonparametric tests in IBM SPSS Statistics 26.0 software. It can be seen from Table 22 that there was no significant difference in the worm reduction rate among groups on the 2nd day; on the 4th day, there were significant differences in the worm reduction rate between the commercially available drug group and the compound juglone ivermectin liniment group and the single-component juglone group, and the P values were all 0.028 (0.01 < P < 0.05); on the 7th day, there were extremely significant differences in the worm reduction rate between the commercially available drug group and the compound juglone liniment group and the single-component juglone group (P < 0.01); on the 14th day, there were extremely significant differences in the worm reduction rate between the commercially available drug group and the compound juglone liniment group and the single-component juglone group, and the P values were all 0.003.

[0193] In the compound juglone ivermectin liniment group, the live mite rate of the experimental animals was 0.5% on the 2nd day, and the negative conversion rate reached 80%. After the second administration, there were no live mites, and there was no recurrence within 14 days, and the negative conversion rate reached 100%. In the single-component juglone group, the live mite rate of the animals was 1.0% after one administration, and the negative conversion rate was 60%. On the 4th day, there were no live mites in the experimental animals of the single-component juglone group, and there was no recurrence within 14 days, and the negative conversion rate reached 100%. In the commercially available drug group, the live mite rate of the experimental animals was 3.9% on the 2nd day. After the second administration, the live mite rate dropped to 0.4%. The live mite rates on the 7th and 14th days were 1.5% and 1% respectively, and the negative conversion rate within 14 days was 50%.

[0194] The results showed that both the compound juglone ivermectin liniment group and the single-component juglone group could effectively treat rabbits with ear mites, and the compound group had a better effect, both being superior to the commercially available drug group.

[0195] b. Lesion score

[0196] Table 23 Rabbit sarcoptic mange infection degree and score (n = 10)

[0197]

[0198] Note: Different lowercase letters indicate significant differences (0.01 < p < 0.05), and different capital letters indicate extremely significant differences p < 0.01

[0199] The experimental record data were analyzed by one-way analysis of variance in IBM SPSS Statistics 26.0 software, and the clinical symptom scores at different times were obtained as shown in Table 23 above. It can be seen from Table 23 that on the 2nd day, there was no significant difference in the animal scores among the compound juglone liniment group, the single-component juglone liniment group, and the control drug group, and the scores of the compound group, the single-component group, and the control drug group were all extremely significantly different from those of the non-drug group; on the 4th, 7th, and 14th days, there was no significant difference in the animal scores between the compound juglone liniment group and the single-component juglone liniment group, and both groups were extremely significantly different from the control drug group.

[0200] The results showed that on the 2nd day, there was no difference in the clinical symptom scores among the compound liniment group, juglone single-drug group and commercially available drug group, and there was a highly significant difference between the three groups and the non-drug group, that is, the three groups all had therapeutic effects on the diseased animals; on the 4th day, there were no live mites in the animals of the compound juglone liniment group and the single juglone group, and there were still live mites in some animals of the commercially available drug group, and there were differences in the clinical symptom scores of the animals in the commercially available group compared with other groups; on the 7th and 14th days, the clinical scores of the animals in the compound drug group, single drug group and commercially available drug group gradually decreased. It can be seen from the results that the compound juglone ivermectin liniment had the best therapeutic effect on rabbit sarcoptic mange, followed by the single juglone group, and the therapeutic effects of both on rabbit ear sarcoptic mange were better than those of the commercially available drug group.

[0201] c. Body weight gain

[0202] Table 24 Record form of body weight change

[0203]

[0204] Note: The same superscripts in the table represent no significant difference, and different superscript letters represent highly significant differences.

[0205] After processing and analyzing the data with IBM SPSS Statistics 26.0, it can be seen from Table 24 that the body weight of the non-drug group animals decreased, while the body weight of the animals in each treatment group showed an overall increasing trend. The body weight gain of the compound juglone ivermectin liniment group after drug treatment was 0.149 kg, the body weight gain of the single juglone liniment group after drug treatment was 0.131 kg, and the body weight gain of the commercially available drug group after treatment was 0.069 kg. There was no significant difference in the body weight gain of each treatment group during the treatment period, and there was a highly significant difference in the body weight gain compared with the non-drug group.

[0206] The above are only the preferred embodiments of the present invention and are not used to limit the present invention. For those skilled in the art, the present invention can have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A compound ivermectin juglone liniment, characterized in that, It consists of the following components by weight percentage: Juglone 0.2% - 0.7% Ivermectin 0.3% - 0.7% Tween - 80 0.8% - 1% Butylated hydroxytoluene 0.1 - 0.15% Azone 0.2% - 5% Menthol 0.05% - 5% Isopropyl myristate 1% - 12% The balance is medium - chain triglyceride.

2. The compound ivermectin juglone liniment according to claim 1, characterized in that, The weight percentages of each component are as follows: Juglone 0.5% Ivermectin 0.5% Tween - 80 1% Butylated hydroxytoluene 0.12% Azone 0.5% - 5% Menthol 0.1% - 5% Isopropyl myristate 2% - 10% The balance is medium - chain triglyceride.

3. The preparation method of the compound ivermectin juglone liniment according to any one of claims 1 to 2, characterized in that, It includes the following steps: Step 1: Mix and stir evenly the formula amounts of butylated hydroxytoluene, juglone, ivermectin, and medium - chain triglyceride, and place them in an ultrasonic bath at 20°C - 30°C for dissolution to obtain a mixed solution; Step 2: Add the formula amounts of triglyceride, azone, menthol, isopropyl myristate, and Tween - 80 to the above - mentioned mixed solution, and vortex thoroughly to obtain the compound ivermectin juglone liniment.

4. The preparation method of the compound ivermectin juglone liniment according to claim 3, characterized in that, In Step 1, the temperature of ultrasonic dissolution is 25°C.

5. The preparation method of the compound ivermectin juglone liniment according to claim 3, characterized in that, The obtained compound ivermectin juglone liniment is stored at 40°C for at least 10 days.

6. Use of the compound ivermectin juglone liniment according to any one of claims 1 - 2, characterized in that The compound ivermectin juglone liniment is applied to the position of the animal skin surface infected with parasites.

7. Use of the compound ivermectin juglone liniment according to claim 6, characterized in that, The animals include cattle, sheep, and rabbits.

8. Use of the compound ivermectin juglone liniment according to claim 6, characterized in that, The parasites include Psoroptes ovis and Sarcoptes scabiei.

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