A ferulaic acid levamisole nano oral liquid, a preparation method and application thereof

Abstract

The application belongs to the technical field of medicine, and particularly relates to a ferulaic acid levamisole nano oral liquid, a preparation method thereof and application. The oral liquid is a water-in-oil-in-water (W1 / O / W2) type double nano emulsion, which is composed of an inner water phase, an intermediate oil phase and an outer water phase. The inner water phase is ferulaic acid levamisole, a first surfactant, a first antioxidant and water for injection. The intermediate oil phase is corn oil, a second surfactant, a third surfactant and a second antioxidant. The outer water phase is a fourth surfactant, a preservative and water for injection. In the application, the inner water phase and the intermediate oil phase are mixed to prepare a W1 / O type primary emulsion, and then the W1 / O type primary emulsion and the outer water phase are mixed to prepare a W1 / O / W2 type multiple emulsion. The method is simple and easy to be industrialized and mass-produced. The ferulaic acid levamisole nano oral liquid has the characteristics of small particle size, high encapsulation rate, good sustained-release performance, good antioxidant property, strong stability and excellent preservative efficacy, and can be used for deworming of pets through a direct oral route, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a ferulic acid levamisole nano-oral solution, its preparation method, and its application. Background Technology

[0002] Parasites such as roundworms, hookworms, and Giardia lamblia that infest the intestines of pets are recognized as important zoonotic pathogens. Their infective eggs, cysts, or larvae can contaminate the environment and spread across hosts through feces, soil, and water, causing diseases such as visceral larval migration syndrome and chronic diarrhea in humans, posing a significant health threat to children and immunocompromised individuals. Therefore, developing highly effective drugs and formulations for the prevention and treatment of pet intestinal parasites is a key intervention strategy to break the pet-to-human transmission chain, and has significant clinical and public health value for protecting the public health and safety of pet owners and promoting the healthy and sustainable development of the pet industry.

[0003] Levamisole ferulic acid is a novel anthelmintic candidate drug. Ferulic acid induces oxidative stress damage in parasites and promotes apoptosis, while simultaneously regulating the host's immune response, enhancing the body's ability to clear pathogens, and alleviating histopathological damage caused by excessive inflammatory activation. Levamisole combines anthelmintic activity with immunomodulatory functions. Its mechanism of action involves selectively inhibiting succinate dehydrogenase activity in parasite muscle tissue, blocking energy metabolism pathways, thereby causing flaccid paralysis and loss of intestinal adhesion ability in the parasite. This compound has good inhibitory and killing effects on various zoonotic parasites such as Ascaris lumbricoides, hookworms, and Toxoplasma gondii, and has been widely used in human and veterinary clinical practice. However, levamisole has poor stability in alkaline environments, is easily degraded and inactivated, and has poor water solubility. Conventional formulations (such as tablets and ordinary oral solutions) cannot overcome these shortcomings, easily leading to problems such as drug precipitation, formulation layering, and insufficient efficacy, thus limiting its clinical application.

[0004] Preparing levamisole ferulic acid nanoparticles can improve drug dissolution and oral bioavailability. Currently, levamisole ferulic acid nanoformulations are mostly produced using oil-in-water (O / W) nano-injection solutions. While this yields nanoemulsions with small particle sizes, it suffers from drawbacks such as demanding preparation conditions, insufficient stability, and invasive delivery methods. Therefore, developing a novel, stable levamisole ferulic acid nanoparticle oral solution capable of achieving sustained-release effects has both clinical significance and market value. Summary of the Invention

[0005] The purpose of this invention is to provide a levamisole ferulic acid nano-oral solution to solve the problem of poor stability of existing levamisole ferulic acid nano-formulations.

[0006] To achieve the above objectives, the present invention provides a levamisole ferulic acid nano-oral solution, wherein the levamisole ferulic acid oral solution comprises an inner aqueous phase, an intermediate oil phase, and an outer aqueous phase, and is composed of the following components by weight percentage: 0.5-1.5% levamisole ferulic acid salt, 4.0-6.0% first surfactant, 0.2-1.0% first antioxidant, 10.0-20.0% corn oil, 0.6-1.0% second surfactant, 0.6-1.0% third surfactant, 0.2-0.8% second antioxidant, 18.0-22.0% fourth surfactant, 0.15-0.25% preservative, and the balance being water for injection; wherein the inner aqueous phase comprises levamisole ferulic acid, the first surfactant, the first antioxidant, and water for injection; the intermediate oil phase comprises corn oil, the second surfactant, the third surfactant, and the second antioxidant; and the outer aqueous phase comprises the fourth surfactant, the preservative, and water for injection.

[0007] This invention also provides a method for preparing the above-mentioned levamisole ferulic acid nano-oral solution, comprising the following steps: S1. Preparation of the internal aqueous phase: Dissolve the first surfactant, levamisole ferulic acid salt and the first antioxidant in water for injection to obtain the internal aqueous phase; S2. Preparation of the oil phase: Mix corn oil, second surfactant, third surfactant and second antioxidant evenly to obtain the oil phase; S3. Preparation of primary emulsion: Under high-speed shearing or ultrasonic conditions, the internal aqueous phase obtained in step S1 is added to the oil phase obtained in step S2, and emulsified for 10-15 min to obtain W1 / O type primary emulsion. S4. Preparation of external aqueous phase: Dissolve the fourth surfactant and preservative in water for injection to obtain the external aqueous phase; S5. Preparation of double emulsion: Under stirring conditions, the W1 / O type primary emulsion obtained in step S3 is added to the external aqueous phase obtained in step S4, and stirred for 15-30 min to obtain the W1 / O / W2 type double emulsion, namely ferulic acid levamisole nano oral solution.

[0008] Optionally, the particle size of the levamisole ferulic acid oral solution is 100–300 nm.

[0009] Optionally, the first and fourth surfactants are one or more of Loxamer 188, Poloxamer 407, and Poloxamer 407; the second surfactant is one or more of Tween 20, Tween 40, and Tween 80; the third surfactant is one or more of Span 40, Span 60, and Span 80; the first antioxidant is vitamin C; the second antioxidant is vitamin E; and the preservative is sodium benzoate. In this formulation, vitamin C serves as the first antioxidant, and vitamin E serves as the second antioxidant, forming a composite antioxidant system. Vitamin C, as a water-soluble antioxidant, preferentially distributes in the aqueous phase, scavenging free radicals in the aqueous phase; vitamin E, as a fat-soluble antioxidant, distributes in the oil phase, protecting corn oil from oxidation. Vitamin C and vitamin E work together to exert antioxidant effects, while simultaneously protecting the drug in the aqueous phase and the oil-water interface film.

[0010] Optionally, in step S3, the high-speed shearing conditions are: rotation speed 8000-12000 rpm, shearing time 10-15 min; the ultrasonic conditions are: probe-type ultrasonic power 200-400 W, ultrasonic time 10-15 min; step S3 is performed in an ice bath.

[0011] Optionally, in step S5, magnetic stirring is used, and the speed is 300-500 rpm.

[0012] Optionally, in step S3, the volume ratio of the internal aqueous phase to the oil phase is 1:1 to 10.

[0013] Optionally, in step S5, the volume ratio of W1 / O type colostrum to external aqueous phase is 1:1 to 10.

[0014] Optionally, the method further includes the following steps: S6, post-processing: filtering the W1 / O / W2 type double emulsion obtained in step S5 using a microporous membrane, adjusting the pH of the filtrate to 3.5-4.5, filling it into a light-proof container, and storing it at 4°C.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. This invention employs a W / O / W dual emulsification method to effectively encapsulate levamisole ferulic acid salt in the inner aqueous phase, reducing the contact between the drug and the external environment, significantly improving the stability of the drug, and preventing drug oxidative degradation. At the same time, nano-sized emulsion droplets can increase the specific surface area of ​​the drug, improve the solubility of the drug, enhance the bioavailability in vivo, achieve slow release of the drug, and prolong the duration of drug efficacy.

[0016] 2. This invention is the first to use corn oil instead of traditional soybean oil as the oil phase to prepare ferulic acid levamisole salt nanoemulsion oral liquid. Corn oil is rich in unsaturated fatty acids and natural vitamin E, has good biocompatibility and safety, and has good applicability in W / O / W dual emulsion systems. It can provide an excellent dissolving carrier for vitamin E, which facilitates the uniform dispersion of vitamin E and the full exertion of its antioxidant function.

[0017] 3. In this invention, poloxamer at different concentrations is used as a surfactant in the inner and outer aqueous phases, and Tween and Span are used as a composite emulsifier in the intermediate oil phase. The hydrophilic-lipophilic balance between the phases can be optimized, thereby forming a stable W / O / W type dual nanoemulsion structure. This solves the problems of poor emulsification effect of a single surface or top layer and easy layering of emulsions. At the same time, poloxamer, Tween and Span are both pharmaceutical grade materials with good biocompatibility and are safe and non-irritating when taken orally.

[0018] 4. The preparation method of the levamisole ferulic acid nano-oral solution in this invention is simple, with controllable parameters and good repeatability. It requires no complex equipment and is suitable for large-scale industrial production. The raw material ratio is scientifically reasonable, and the prepared nano-oral solution has uniform particle size, strong stability, low irritation, and is convenient to take, effectively solving the defects of existing levamisole ferulic acid salt nano-preparations. It can be used to deworm pets through direct oral administration to treat and prevent intestinal roundworms, hookworms, whipworms, coccidia, Toxoplasma gondii, and strongyloides stercoralis, etc. It is convenient to use, simple in process, suitable for large-scale industrial production, and has a very broad application prospect. Attached Figure Description

[0019] Figure 1 This is a particle size distribution diagram of the nanoemulsion in Example 4 of the present invention; Figure 2 This is a microscope image of the nanoemulsion particles in Example 4 of the present invention. Detailed Implementation

[0020] The present invention will now be described in further detail with reference to the accompanying drawings, so that those skilled in the art can implement it based on the description. The following description is intended to disclose the present invention and enable those skilled in the art to implement it. The preferred embodiments described below are merely examples, and those skilled in the art can conceive of other obvious modifications. The basic principles of the invention defined in the following description can be applied to other embodiments, modifications, improvements, equivalents, and other technical solutions that do not depart from the spirit and scope of the invention. In this invention, unless otherwise specified, the raw materials and equipment used are commercially available or commonly used in the art. The methods in the following embodiments, unless otherwise specified, are conventional methods in the art.

[0021] This invention provides a levamisole ferulic acid nano-oral solution, which comprises an inner aqueous phase, an intermediate oil phase and an outer aqueous phase, with a particle size of 100-300 nm, as shown in Table 1 by weight percentage.

[0022] Table 1 Composition of Ferulic Acid Levamisole Nano Oral Solution

[0023] In Table 1, "to bring to 100%" means: the weight percentage of the inner aqueous phase + the weight percentage of the intermediate oil phase + the weight percentage of the outer aqueous phase = 100%. Any deficiency is made up with water for injection in the outer aqueous phase. The first and fourth surfactants are one or more of poloxamer 188, poloxamer 407, and poloxamer 407; the second surfactant is one or more of Tween 20, Tween 40, and Tween 80; and the third surfactant is one or more of Span 40, Span 60, and Span 80. The first antioxidant is vitamin C, and the second antioxidant is vitamin E. The preservative is sodium benzoate, preferably used at a concentration of 0.1–0.25%. Sodium benzoate, as a preservative, is distributed in the outer aqueous phase. Under acidic conditions (below pH 4), it can be converted into benzoic acid, which has antibacterial activity, inhibiting molds, yeasts, and some bacteria, effectively preventing microbial contamination of the formulation during storage.

[0024] The preparation method of the above-mentioned levamisole ferulic acid nano-oral solution is as follows: S1. Preparation of the internal aqueous phase: Dissolve the first surfactant, levamisole ferulic acid salt and the first antioxidant in water for injection to obtain the internal aqueous phase.

[0025] S2. Preparation of the oil phase: Mix corn oil, second surfactant, third surfactant and second antioxidant evenly to obtain the oil phase.

[0026] S3. Preparation of primary emulsion: Under high-speed shearing or ultrasonic conditions, the internal aqueous phase obtained in step S1 is added to the oil phase obtained in step S2, with a volume ratio of internal aqueous phase to oil phase of 1:1 to 10. Emulsification is carried out for 10 to 15 min to obtain W1 / O type primary emulsion. Among them, the high-speed shearing conditions are: rotation speed 8000 to 12000 rpm, shearing time 10 to 15 min; ultrasonic conditions are: probe ultrasonic power 200 to 400 W, ultrasonic time 10 to 15 min; the whole process is carried out in an ice bath.

[0027] S4. Preparation of external aqueous phase: Dissolve the fourth surfactant and preservative in water for injection to obtain the external aqueous phase.

[0028] S5. Preparation of double emulsion: Under stirring conditions, the W1 / O type primary emulsion obtained in step S3 is added to the external aqueous phase obtained in step S4. The volume ratio of W1 / O type primary emulsion to external aqueous phase is 1:1 to 10. The mixture is magnetically stirred for 15 to 30 minutes at a speed of 300 to 500 rpm to obtain W1 / O / W2 type double emulsion, namely ferulic acid levamisole nano oral solution.

[0029] S6. Post-processing: Filter the W1 / O / W2 type double emulsion obtained in step S5 using a microporous membrane, adjust the pH of the filtrate to 3.5-4.5, fill it into a light-proof container, and store it at 4°C.

[0030] Example 1 This embodiment provides a levamisole ferulic acid nano-oral solution, the preparation method of which includes the following steps: (1) Preparation of internal aqueous phase: Weigh 4.0 g poloxamer 188, 0.5 g levamisole ferulic acid salt and 0.2 g vitamin C, add water for injection to a final volume of 50 mL, stir and dissolve at 40 °C to obtain internal aqueous phase (W1).

[0031] (2) Preparation of oil phase: Weigh 20.0 g corn oil, add 1.0 g Tween 80, 1.0 g Span 80 and 0.2 g vitamin E, stir and mix evenly to obtain oil phase (O).

[0032] (3) Preparation of colostrum: Under probe-type ultrasound (power 300 W) conditions, keep the temperature low in an ice bath, slowly add the inner aqueous phase (W1) to the oil phase (O), and ultrasonically emulsify for 15 min to obtain W1 / O type colostrum.

[0033] (4) Preparation of external aqueous phase: Weigh 20.0 g of poloxamer 188 and 0.2 g of sodium benzoate, add an appropriate amount of water for injection, stir to dissolve, and then dilute to a total of 100 g with water for injection (this refers to the total of 100 g of poloxamer 188, sodium benzoate and water for injection in the internal aqueous phase, intermediate oil phase and external aqueous phase), to obtain the external aqueous phase (W2).

[0034] (5) Preparation of double emulsion: Under magnetic stirring (200 rpm), the W1 / 0 type primary emulsion was slowly added dropwise to the external aqueous phase (W2), and stirring was continued for 20 min to obtain the W1 / 0 / W2 type double emulsion.

[0035] (6) Homogenization and granulation: The temperature was controlled at ≤40℃ in an ice-water bath, and the pressure was 90 MPa. The homogenization was repeated 8 times to obtain a light yellow semi-transparent nanoemulsion.

[0036] (7) Post-processing: The pale yellow semi-transparent nano-emulsion is cooled to room temperature, filtered through a 0.45μm filter membrane, and the pH is adjusted to 4.0 with an appropriate amount of hydrochloric acid solution. It is then aseptically filled into brown bottles and sealed to obtain the ferulic acid levamisole nano oral liquid product.

[0037] Example 2 This embodiment provides a ferulic acid levamisole nano-oral solution, which is prepared in the same way as the preparation method in Example 1, except that: in this embodiment, the amount of poloxamer 188 in the inner aqueous phase (W1) is 5 g, the amount of ferulic acid levamisole salt is 1 g, and the amount of vitamin C is 0.5 g; the amount of corn oil in the oil phase (O) is 15 g, the amount of Tween 80 is 0.8 g, the amount of Span 80 is 0.8 g, and the amount of vitamin E is 0.5 g; the ultrasound time of the probe is 12 min; the amount of poloxamer 188 in the outer aqueous phase (W2) is 18 g, the amount of sodium benzoate is 0.15 g, and the stirring time in step (5) is 25 min; the pH is adjusted to 3.8 in the post-treatment.

[0038] Example 3 This embodiment provides a ferulic acid levamisole nano-oral solution, which is prepared in the same way as the preparation method in Example 1, except that: in this embodiment, the amount of poloxamer 188 in the inner aqueous phase (W1) is 6.0 g, the amount of ferulic acid levamisole salt is 1.5 g, and the amount of vitamin C is 0.8 g; the amount of corn oil in the oil phase (O) is 10 g, the amount of Tween 80 is 0.6 g, the amount of Span 80 is 0.6 g, and the amount of vitamin E is 0.8 g; the ultrasound time of the probe is 10 min; the amount of poloxamer 188 in the outer aqueous phase (W2) is 22 g, the amount of sodium benzoate is 0.25 g, and the stirring time in step (5) is 30 min; the pH is adjusted to 4.2 in the post-treatment.

[0039] The composition of the ferulic acid levamisole nano oral solution in Examples 1-3 is shown in Table 2.

[0040] Table 2 Composition of Ferulic Acid Levamisole Nano Oral Solution

[0041] In Table 2, "total amount of 100 g" means: mass of internal aqueous phase + mass of intermediate oil phase + mass of external aqueous phase = 100 g, and any shortfall is made up by water for injection in the external aqueous phase.

[0042] Experimental Example The ferulic acid levamisole nano-oral solution prepared in Example 1 was subjected to particle size analysis using a dynamic light scattering (DLS) particle size analyzer and an optical microscope. The results are as follows: Figure 1 and Figure 2 As shown. By Figure 1 and Figure 2 It is known that the particle size of the W1 / 0 / W2 type nanoemulsion particles prepared by the present invention is 213±3.1 nm.

[0043] For the levamisole ferulic acid nano-oral solutions prepared in Examples 1-3, their particle size and drug loading were measured at different time periods, and the results are shown in Table 3.

[0044] Table 3. Particle size and drug loading of levamisole ferulic acid nano-oral solution products in Examples 1-3 at different time points.

[0045] As shown in Table 3, the levamisole ferulic acid nano-oral solution products in Examples 1-3 showed no aggregation or stratification within 90 days, and the nanoparticle size and drug loading did not change significantly. This indicates that the W1 / 0 / W2 type nanoemulsion prepared by this invention has good stability and water solubility, and the particle size is stable and uniform, which improves the effective storage time of levamisole ferulic acid and has a very broad application prospect.

[0046] The above are merely embodiments of the present invention. The invention is not limited to the fields covered by these embodiments. Commonly known structures and characteristics are not described in detail here. Those skilled in the art are aware of all common technical knowledge in the field prior to the application date or priority date, are aware of all existing technologies in that field, and have the ability to apply conventional experimental methods prior to that date. Those skilled in the art can improve and implement this solution based on the guidance provided in this application and their own capabilities. Some typical known structures or methods should not be obstacles for those skilled in the art to implement this application. It should be noted that those skilled in the art can make several modifications and improvements without departing from the structure of the present invention. These should also be considered within the scope of protection of the present invention, and will not affect the effectiveness or practicality of the invention. The scope of protection claimed in this application should be determined by the content of its claims. The specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A ferulic acid levamisole nano-oral solution, characterized in that: The levamisole ferulic acid oral solution comprises an inner aqueous phase, an intermediate oil phase, and an outer aqueous phase, and is composed of the following components by weight percentage: 0.5-1.5% levamisole ferulic acid salt, 4.0-6.0% first surfactant, 0.2-1.0% first antioxidant, 10.0-20.0% corn oil, 0.6-1.0% second surfactant, 0.6-1.0% third surfactant, 0.2-0.8% second antioxidant, 18.0-22.0% fourth surfactant, 0.15-0.25% preservative, and the balance being water for injection; wherein, the inner aqueous phase consists of levamisole ferulic acid, the first surfactant, the first antioxidant, and water for injection; the intermediate oil phase consists of corn oil, the second surfactant, the third surfactant, and the second antioxidant; and the outer aqueous phase consists of the fourth surfactant, the preservative, and water for injection.

2. The ferulic acid levamisole nano-oral solution according to claim 1, characterized in that: The particle size of the levamisole ferulic acid oral solution is 100–300 nm.

3. The ferulic acid levamisole nano-oral solution according to claim 1, characterized in that: The first and fourth surfactants are one or more of poloxamer 188, poloxamer 407, and poloxamer 407; the second surfactant is one or more of Tween 20, Tween 40, and Tween 80; the third surfactant is one or more of Span 40, Span 60, and Span 80; the first antioxidant is vitamin C; the second antioxidant is vitamin E; and the preservative is sodium benzoate.

4. The method for preparing the ferulic acid levamisole nano-oral solution according to any one of claims 1-3, characterized in that: Includes the following steps: S1. Preparation of the internal aqueous phase: Dissolve the first surfactant, levamisole ferulic acid salt and the first antioxidant in water for injection to obtain the internal aqueous phase; S2. Preparation of the oil phase: Mix corn oil, second surfactant, third surfactant and second antioxidant evenly to obtain the oil phase; S3. Preparation of primary emulsion: Under high-speed shearing or ultrasonic conditions, the internal aqueous phase obtained in step S1 is added to the oil phase obtained in step S2, and emulsified for 10-15 min to obtain W1 / O type primary emulsion. S4. Preparation of external aqueous phase: Dissolve the fourth surfactant and preservative in water for injection to obtain the external aqueous phase; S5. Preparation of double emulsion: Under stirring conditions, the W1 / O type primary emulsion obtained in step S3 is added to the external aqueous phase obtained in step S4, and stirred for 15-30 min to obtain the W1 / O / W2 type double emulsion, namely ferulic acid levamisole nano oral solution.

5. The method for preparing ferulic acid levamisole nano-oral solution according to claim 4, characterized in that: In step S3, the high-speed shearing conditions are: rotation speed 8000-12000 rpm, shearing time 10-15 min; the ultrasonic conditions are: probe-type ultrasonic power 200-400 W, ultrasonic time 10-15 min; step S3 is performed in an ice bath.

6. The method for preparing levamisole ferulic acid nano-oral solution according to claim 4, characterized in that: In step S5, magnetic stirring is used, with a speed of 300-500 rpm.

7. The method for preparing levamisole ferulic acid nano-oral solution according to claim 4, characterized in that: In step S3, the volume ratio of the internal aqueous phase to the oil phase is 1:1 to 10.

8. The method for preparing levamisole ferulic acid nano-oral solution according to claim 4, characterized in that: In step S5, the volume ratio of W1 / O type colostrum to external aqueous phase is 1:1 to 10.

9. The method for preparing ferulic acid levamisole nano-oral solution according to claim 4, characterized in that: The process also includes the following steps: S6, post-processing: the W1 / O / W2 type double emulsion obtained in step S5 is filtered through a microporous membrane, the pH of the filtrate is adjusted to 3.5-4.5, and the solution is filled into a light-proof container and stored at 4°C.

10. The use of levamisole ferulic acid nano-oral solution as described in any one of claims 1-3 in the preparation of immune-enhancing drugs or pet deworming drugs.

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