Albendazole-ivermectin nanoemulsion and preparation method thereof

Abstract

The invention discloses an albendazole-ivermectin compound nanoemulsion and a preparation method of the albendazole-ivermectin compound nanoemulsion. The albendazole-ivermectin nanoemulsion is prepared from albendazole, ivermectin, an oil phase, a surfactant, a cosurfactant and a water phase. The preparation method comprises the following steps: dissolving the medicines in a solvent, extracting to obtain an oil phase, removing the solvent, adding the oil phase into a water phase consisting of the surfactant, the cosurfactant and water under stirring, and spontaneously forming the nano-emulsion through low-energy emulsification. The prepared nanoemulsion is small in particle size and good in stability, the solubility and the drug loading capacity of two insoluble drugs can be remarkably improved, the synergistic parasite expelling effect is achieved, and the cure rate is increased. The preparation process is simple, mild in condition and suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of veterinary preparations, in particular to a compound nanoemulsion preparation containing albendazole and ivermectin and a preparation method thereof. BACKGROUND

[0002] Albendazole is a highly effective broad-spectrum benzimidazole anthelmintic drug, which has good killing effect on nematodes, tapeworms and flukes. Ivermectin is a macrolide antiparasitic drug, which has excellent activity against endo- and ectoparasites, especially arthropods and nematodes. The combination of the two can broaden the spectrum of activity, produce synergistic effects, improve the anthelmintic effect, and help delay the development of parasitic drug resistance.

[0003] However, albendazole and ivermectin are both poorly soluble drugs, and their low bioavailability restricts the clinical efficacy. Conventional preparations such as tablets, powders and ordinary solutions have problems such as poor solubility, poor stability, incomplete absorption and inconvenience of use.

[0004] Nanoemulsion is a thermodynamically stable colloidal dispersion system composed of oil phase, water phase, surfactant and co-surfactant, with a particle size of usually 10-200 nm. As a non-equilibrium system, nanoemulsion cannot form spontaneously. Therefore, the input of energy from mechanical devices or chemical reactions is indispensable. It can be divided into high-energy emulsification and low-energy emulsification by different energy supply methods. As a drug carrier, it can significantly improve the solubility and bioavailability of poorly soluble drugs, improve drug stability, delay drug release, reduce drug side effects, and achieve various administration routes such as oral and injection.

[0005] At present, there is no report on the preparation of a compound nanoemulsion preparation of albendazole and ivermectin. SUMMARY

[0006] The present application aims to overcome the shortcomings of the prior art and provide a compound nanoemulsion preparation capable of significantly improving the solubility, bioavailability and stability of albendazole and ivermectin.

[0007] Technical solution: A albendazole-ivermectin compound nanoemulsion is prepared from the following components by weight percentage: Albendazole: 0.1%-5% Ivermectin: 0.1%-2% Oil phase: 5%-30% Surfactant: 15%-40% Co-surfactant: 5%-25% The balance is water phase, supplemented to 100% Preferably, the oil phase is selected from one of isopropyl myristate, isopropyl palmitate, ethyl oleate, and 0.5%-5% cetyl alcohol by weight of the oil phase.

[0008] Preferably, the surfactant is selected from one of polysorbate-80 (Tween-80), polyoxyethylene castor oil (Cremophor EL), polyethylene glycol-8-glyceryl caprylate / caprate (Labrasol).

[0009] Preferably, the co-surfactant is selected from one of anhydrous ethanol, polyethylene glycol-400 (PEG-400), 1,2-propanediol.

[0010] The preparation method of the albendazole-ivermectin compound nanoemulsion comprises the following steps: Dissolution of the drug: dissolve albendazole and ivermectin in dimethyl sulfoxide (DMSO) solution according to the prescription amount, heat to 40-60℃ and stir to dissolve, obtain an organic phase solution, the amount of DMSO is 2-4 times the mass of albendazole and ivermectin; Solubilization of the drug: add the oil phase according to the prescription amount to the prepared organic phase solution, keep the system temperature at 40-60℃, and obtain a mixed solution; Preparation of the oil phase solution: wash the prepared mixed solution with purified water at a weight ratio of 2-5 times, stir uniformly, then stand and separate, discard the water phase, and repeat the purified water washing operation for the remaining oil phase until the DMSO content in the oil phase is not more than 0.3%, and obtain the oil phase solution; Preparation of the water phase: mix the surfactant and co-surfactant according to the prescription amount, then add the water phase according to the prescription amount, stir uniformly, and obtain a uniform water phase solution; Initial formation of the nanoemulsion: slowly add the oil phase solution to the water phase solution under continuous stirring (for example, 300-600 rpm), and stir until a clear and transparent nanoemulsion is formed; Beneficial effects: (1) High drug loading and solubility: through the nanoemulsion system, the two poorly soluble drugs albendazole and ivermectin are simultaneously wrapped in the oil phase inner core, which significantly improves their apparent solubility and drug loading of the preparation. The solubility in the aqueous phase system increases from extremely low to nanometer solubility. In vivo tests have confirmed that under the same dosage, the prepared nanoemulsion has better in vivo deworming effect, and the albendazole-ivermectin nanoemulsion treatment group can achieve a worm egg reduction rate of 93.2% after 10 days of administration, which is much higher than the 53.4% of the albendazole-ivermectin premix treatment group.

[0011] (2) Good stability: the nanoemulsion prepared by the scheme is a thermodynamic stable system, and the stability is better than that of ordinary emulsion and premix. The nanoemulsion prepared by adding cetyl alcohol in the formula can eliminate austenitic aging, and the nanoemulsion is stable under centrifugation, and the properties do not change during long-term stability.

[0012] (3) Convenient to use: the preparation is a liquid preparation, which can be dissolved in water, and uniform administration can be achieved when administered to a group, thereby avoiding poor efficacy caused by uneven administration.

[0013] (4) Simple process: the preparation process does not require complex equipment, and a simple low-energy emulsification method can be used, which is mild and suitable for industrial large-scale production. BRIEF DESCRIPTION OF DRAWINGS The application will be further described in detail below with reference to the accompanying drawings and specific implementation methods Figure 1 The transmission electron microscope photo of the nanoemulsion prepared for implementation 1; Figure 2 The actual photo of the nanoemulsion prepared for implementation 1; DETAILED DESCRIPTION

[0014] The application will be further described in detail below with reference to the accompanying drawings and specific implementation methods

[0015] Example 1 An albendazole-ivermectin compound nanoemulsion is composed of the following components: Albendazole: 2.0 g Ivermectin: 0.5 g Isopropyl myristate (IPM): 15.0 g Cetyl alcohol: 0.15 g Polysorbate-80 (Tween-80): 30.0 g Anhydrous ethanol: 15.0 g Purified water: 37.35 g Total: 100.0 g Preparation method: Dissolve albendazole and ivermectin in 7.5 g DMSO solution according to the formula amount, heat to 50℃ and stir to dissolve, to obtain an organic phase solution.

[0016] Add IPM 15.0 g and cetyl alcohol 0.15 g to the above organic phase solution, keep the system temperature at 50℃, to obtain a mixed solution; To the above mixed solution, 55.5 g purified water was added, and stirred for about 10 min until uniform, then layered after standing, the water phase was discarded, and the oil phase was washed with purified water again for two times. After the end of washing, the content of DMSO in the oil phase was 0.06% detected by gas chromatography, and the oil phase solution was obtained; 30.0 g Tween-80 and 15.0 g anhydrous ethanol were mixed uniformly, then 37.35 g purified water was added, stirred and mixed uniformly to obtain a uniform aqueous phase solution; The oil phase solution was slowly added to the aqueous phase solution under stirring at 450 rpm, and stirring was continued for 40 min to form a clear and transparent nanoemulsion.

[0017] It was detected that the obtained nanoemulsion was clear and yellowish in appearance. The average particle size was 23.3 nm and the PDI was 0.152 as determined by a laser particle size instrument. Transmission electron microscopy observation showed that the particles were regular spherical and uniformly distributed, as shown in the following figure: Figure 1 It could be infinitely diluted with purified water, and the methylene blue diffusion speed was greater than that of Sudan red, indicating that the prepared nanoemulsion was O / W type.

[0018] Example 2 An albendazole-ivermectin compound nanoemulsion was composed of the following components: Albendazole: 2.0 g Ivermectin: 0.5 g Isopropyl palmitate: 20.0 g Cetyl alcohol: 1.0 g Polyoxyethylene castor oil (Cremophor EL): 20.0 g Polyethylene glycol-400 (PEG-400): 15.0 g Purified water: 41.5 g Total: 100.0 g Preparation method: Albendazole and ivermectin were dissolved in 7.5 g DMSO solution according to the formula amount, and the temperature was increased to 50°C for stirring and dissolving until clear to obtain an organic phase solution.

[0019] To the above organic phase solution, 20.0 g isopropyl palmitate and 1.0 g cetyl alcohol were added to keep the system temperature at 60°C to obtain a mixed solution; To the above mixed solution, 105.0 g purified water was added, and stirred for about 12 min until uniform, then layered after standing, the water phase was discarded, and the oil phase was washed with purified water again for two times. After the end of washing, the content of DMSO in the oil phase was 0.03% detected by gas chromatography, and the oil phase solution was obtained; Mix 20.0 g Cremophor EL and 15.0 g PEG-400 homogeneously, then add 41.5 g purified water, stir to mix homogeneously to obtain a uniform aqueous phase solution; Slowly drop the oil phase solution into the aqueous phase solution under 600 rpm stirring, continue stirring for 30 min to form a clear and transparent nanoemulsion.

[0020] It is detected that the obtained nanoemulsion is clear in appearance, with an average particle size of 52.1 nm and a PDI of 0.168.

[0021] Example 3 An albendazole-ivermectin compound nanoemulsion is composed of the following components: Albendazole: 2.0 g Ivermectin: 0.5 g Ethyl oleate: 15.0 g Cetyl alcohol: 1.0 g Polyethylene glycol-8-glyceryl caprylate / caprate (Labrasol): 20.0 g 1,2-Propanediol: 15.0 g Purified water: 46.5 g Total: 100.0 g Preparation method Dissolve albendazole and ivermectin in 7.5 g DMSO solution according to the formulation amount, heat to 50°C and stir to dissolve, to obtain an organic phase solution.

[0022] Add ethyl oleate 15.0 g and cetyl alcohol 1.0 g to the above organic phase solution, keep the system temperature at 60°C to obtain a mixed solution; Add 36.0 g purified water to the above mixed solution, stir for about 14 min until uniform, then stand to separate into layers, discard the aqueous phase, and wash the oil phase with purified water twice. After washing, gas chromatography detects that the DMSO content in the oil phase is 0.11%, to obtain an oil phase solution; Mix 20.0 g Labrasol and 15.0 g 1,2-propanediol homogeneously, then add 46.5 g purified water, stir to mix homogeneously to obtain a uniform aqueous phase solution; Slowly drop the oil phase solution into the aqueous phase solution under 400 rpm stirring, continue stirring for 29 min to form a clear and transparent nanoemulsion.

[0023] It is detected that the obtained nanoemulsion is clear in appearance, with an average particle size of 68.5 nm and a PDI of 0.189.

[0024] Example 4 The difference between the formulation of this example and Example 1 is that no cetyl alcohol is added in the formulation, and the preparation method is the same as that of Example 1.

[0025] Example 5 An albendazole-ivermectin compound nanoemulsion consists of the following components: Albendazole: 5.0 g Ivermectin: 2.0 g Isopropyl myristate (IPM): 30.0 g Cetyl alcohol: 1.5 g Polyoxyethylene castor oil (Cremophor EL): 40.0 g Anhydrous ethanol: 5.0 g Purified water: 26.5 g Total: 100.0 g Preparation method: Dissolve albendazole and ivermectin in 28.0 g DMSO solution according to the formulation amount, and stir to dissolve at 60°C to obtain an organic phase solution.

[0026] Add IPM 30.0 g and cetyl alcohol 1.5 g to the above organic phase solution, and keep the system temperature at 60°C to obtain a mixed solution; Add 157.5 g purified water to the above mixed solution, stir for about 20 min until uniform, then stand to separate the layers, discard the water phase, and wash the oil phase with purified water twice. After washing, the DMSO content in the oil phase is 0.29% detected by gas chromatography, and an oil phase solution is obtained; Mix 40.0 g polyoxyethylene castor oil (Cremophor EL) and 5.0 g anhydrous ethanol uniformly, then add 26.5 g purified water, and stir to mix uniformly to obtain a uniform aqueous phase solution; Slowly add the oil phase solution to the aqueous phase solution under stirring at 300 rpm, continue stirring for 50 min, and form a clear and transparent nanoemulsion.

[0027] The detected nanoemulsion has a clear appearance, an average particle size of 94.6 nm, and a PDI of 0.197.

[0028] Example 6 An albendazole-ivermectin compound nanoemulsion consists of the following components: Albendazole: 0.1 g Ivermectin: 0.1 g Isopropyl myristate (IPM): 5.0 g Cetyl alcohol: 25 mg Polysorbate-80 (Tween-80): 15.0 g Anhydrous ethanol: 5.0 g Purified water: 74.8 g Total: 100.0 g Preparation method: Dissolve albendazole and ivermectin in 0.4 g DMSO solution according to the formula, heat to 50°C and stir until dissolved, to obtain an organic phase solution.

[0029] Add 5.0 g IPM and 25 mg cetyl alcohol to the above organic phase solution, and keep the system temperature at 50°C to obtain a mixed solution; Add 10.0 g purified water to the above mixed solution, stir for about 10 min until uniform, then stand and separate the layers, discard the water phase, and repeat the washing with purified water twice. After washing, the DMSO content in the oil phase is 0.02% as detected by gas chromatography, to obtain an oil phase solution; Mix 15.0 g Tween-80 and 5.0 g anhydrous ethanol uniformly, then add 74.8 g purified water and stir to mix uniformly, to obtain a uniform aqueous phase solution; Slowly add the oil phase solution to the aqueous phase solution under stirring at 300 rpm, continue stirring for 50 min, and form a clear and transparent nanoemulsion.

[0030] The test shows that the obtained nanoemulsion is clear in appearance, with an average particle size of 42.8 nm and a PDI of 0.139. Transmission electron microscopy observation shows that the particles are regular spherical and uniformly distributed.

[0031] Stability evaluation Centrifugation test: centrifuge the prepared compound nanoemulsion at 4000 r / min for 30 min, and observe the appearance change of the compound nanoemulsion, whether turbidity, layering, and drug precipitation occur, etc. The results are shown in the table below:

[0032] As can be seen from the above table, the nanoemulsion prepared in Example 4 appears turbid after centrifugation, indicating that the prepared nanoemulsion preparation is unstable, and both the emulsion and the suspension are unstable. The difference between Example 4 and other examples is that cetyl alcohol is not added in the formula, which indicates that the use of the compound oil phase in the present formula can eliminate Ostwald ripening, and the prepared nanoemulsion is stable under centrifugation.

[0033] Long-term test: store the compound nanoemulsion prepared in Example 1 at a temperature of 25°C±2°C and a relative humidity of 60%±10% for 12 months, and take samples at 0, 3, 6, 9, and 12 months to observe whether the appearance of the nanoemulsion changes, and to measure the drug content change. The results are shown in the table below:

[0034] From the above table, it can be seen that the compound nanoemulsion prepared by the scheme of the present application has no change in appearance, is not layered and demulsified, and the drug components are stable and have no obvious change in content after being stored for 12 months under the condition of temperature 25℃±2℃ and relative humidity 60%±10%. The solubility of albendazole in water is <0.1 mg / L, and the solubility of ivermectin in water is about 4 μg / mL, which is far lower than the concentration in the nanoemulsion of the present scheme. By the nanoemulsion system, the two poorly soluble drugs are simultaneously wrapped in the oil phase inner core, which significantly improves their apparent solubility and drug loading capacity of the preparation.

[0035] In vivo test Healthy pigs that have not used anthelmintics are infected with the laboratory cultured infective stage eggs of Ascaris suum by oral gavage when the pigs are 2-3 weeks after weaning and have a body weight of 10-15 kg.

[0036] The successfully infected pigs are randomly divided into the following groups, with 20 pigs in each group: Group 1: albendazole-ivermectin nanoemulsion treatment group Group 2: albendazole-ivermectin premix treatment group Group 3: infection without drug administration control group After 21 days of infection, the drugs are administered continuously for 7 days, and the McMaster counting method is used to quantitatively calculate the number of worm eggs per gram of feces on day 10, and the reduction rate of fecal worm eggs is calculated.

[0037] Worm egg reduction rate = [(average EPG before administration - average EPG after administration) / average EPG before administration] × 100% Note: PEG is the number of worm eggs per gram of feces

[0038] The results of the present test show that the albendazole-ivermectin nanoemulsion treatment group and the albendazole-ivermectin premix treatment group have anthelmintic effects on Ascaris suum in pigs, and the albendazole-ivermectin nanoemulsion treatment group can achieve a worm egg reduction rate of 93.2% after 10 days of administration, which is much higher than the worm egg reduction rate of 53.4% of the albendazole-ivermectin premix treatment group.

[0039] Of course, the above description is not a limitation on the present application, and the present application is not limited to the above examples only. Changes, modifications, additions or substitutions made by those of ordinary skill in the art within the essential scope of the present application also belong to the protection scope of the present application.

Claims

1. An albendazole-ivermectin compound nanoemulsion, characterized in that, It consists of the following components by weight percentage: albendazole 0.1%-5%, ivermectin 0.1%-2%, oil phase 5%-30%, surfactant 15%-40%, co-surfactant 5%-25%, and the balance being an aqueous phase.

2. The compound nanoemulsion according to claim 1, characterized in that, The oil phase is selected from one of isopropyl myristate, isopropyl palmitate, and ethyl oleate, and cetyl alcohol accounting for 0.5%-5% of the oil phase weight.

3. The compound nanoemulsion according to claim 1, characterized in that, The surfactant is selected from one of polysorbate-80, polyoxyethylene castor oil, and polyethylene glycol-8-glycerol caprylic / decanoic acid ester.

4. The compound nanoemulsion according to claim 1, characterized in that, The co-surfactant is selected from anhydrous ethanol, polyethylene glycol-400, and 1,2-propanediol.

5. The compound nanoemulsion according to any one of claims 1-4, characterized in that, The average particle size of the nanoemulsion is between 10 and 200 nm, and the polydispersity index (PDI) is less than 0.

3.

6. A method for preparing the albendazole-ivermectin compound nanoemulsion as described in any one of claims 1-5, characterized in that, Includes the following steps: a) Dissolve albendazole and ivermectin in dimethyl sulfoxide (DMSO) solution according to the formula amount, heat to 40-60℃ and stir until dissolved to obtain an organic phase solution; b) Add the formulated amount of oil phase to the organic phase solution prepared in step a), and maintain the system temperature at 40-60℃ to obtain a mixed solution; c) Add 2-5 times the weight of purified water to the mixed solution prepared in step b) and wash. After stirring evenly, let it stand to separate into layers. Discard the aqueous phase. The remaining oil phase can be washed with purified water repeatedly to obtain an oil phase solution. d) Mix the formulated amounts of surfactant and co-surfactant with the aqueous phase to obtain an aqueous solution; e) Under stirring conditions, slowly add the oil phase solution obtained in step c) to the aqueous phase solution obtained in step d), and continue stirring until a clear and transparent nanoemulsion is formed.

7. The method according to claim 6, characterized in that, In step a), the amount of DMSO used is 2-4 times the combined mass of albendazole and ivermectin.

8. The method according to claim 6, characterized in that, Step c) The DMSO content of the oil phase after washing does not exceed 0.3%.

9. The method according to claim 6, characterized in that, The stirring speed in step e) is 300-600 rpm.

10. The use of the albendazole-ivermectin compound nanoemulsion according to any one of claims 1-5 in the preparation of a drug for preventing and treating internal and external parasitic diseases in animals, wherein the animals are mammals, including cattle, sheep, pigs, dogs, and cats.

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