Preparation equipment and process of zanamivir biological oral medicine

Abstract

The invention discloses preparation equipment and a process of a zanamivir biological oral medicine. The zanamivir biological oral medicine is characterized by being prepared from the following raw materials in parts by weight: 5-15 mg / mL of zanamivir, 120 mg of glycerin monostearate, 40 mg of soybean phospholipid, 1.6% (w / v) of a praloxamer 188 solution, a proper amount of dichloromethane and 5% of mannitol. Melting glyceryl monostearate and soybean lecithin in dichloromethane to form an oil phase; preparing a zanamivir solution as an internal water phase; adding the inner water phase into the oil phase, and performing dispersion ultrasonic treatment to form W / O primary emulsion; adding the primary emulsion into a praloxamer 188 solution, and performing dispersion ultrasonic treatment again to form W / O / W multiple emulsion; volatilizing the organic solvent to obtain a nano suspension; and adding mannitol, and freeze-drying to obtain the freeze-dried powder. By optimizing the concentration of the internal water phase and the volume of the external water phase, the nanoparticles with uniform particle size and good stability are prepared, the oral bioavailability of zanamivir is remarkably improved, the patient compliance is enhanced, and the zanamivir nano-particles are suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of biological oral drug preparation equipment, and in particular to a preparation equipment and process for the biological oral drug zanamivir. Background Art

[0002] Zanamivir is a neuraminidase inhibitor with significant inhibitory effects against influenza A and B viruses. However, due to its poor water solubility and low oral bioavailability, zanamivir is traditionally administered via inhalation, which presents challenges such as poor patient compliance and inconvenient administration. Therefore, developing a novel formulation that can effectively enhance the oral bioavailability of zanamivir is of great significance. Summary of the Invention

[0003] The technical problem to be solved by the present invention is to provide a preparation device and process for a zanamivir oral biological drug, which significantly improves its oral bioavailability and enhances patient compliance by optimizing the prescription process, and is suitable for industrial production.

[0004] The present invention adopts the following technical solutions to achieve the invention objectives: Disclosed is a preparation device and process for a zanamivir oral biological drug, which is prepared from the following raw materials in parts by weight: 5-15 mg / mL of zanamivir, 120 mg of glyceryl monostearate, 40 mg of soybean lecithin, 1.6% (w / v) of ploxamer 188 solution, an appropriate amount of dichloromethane, and 5% of mannitol.

[0005] As a further limitation of the present technical solution, glyceryl monostearate and soybean lecithin are melted in dichloromethane to form an oil phase; a zanamivir solution is prepared as the internal aqueous phase; the internal aqueous phase is added to the oil phase and dispersed and ultrasonically formed into a W / O colostrum; the colostrum is added to a Ploxamer 188 solution and dispersed and ultrasonically formed again into a W / O / W emulsion; the organic solvent is evaporated to obtain a nanosuspension; mannitol is added, and the mixture is freeze-dried to obtain a lyophilized powder.

[0006] As a further limitation of the present technical solution, the concentration of zanamivir in the inner aqueous phase is 5 mg / mL, 10 mg / mL or 15 mg / mL; the volume of the outer aqueous phase is 4 mL, 8 mL or 12 mL.

[0007] As a further limitation of the present technical solution, the particle size of the nanoparticles is controlled between 150-500 nm, the surface potential is between -45 and -55 mV, the encapsulation efficiency is ≥30%, and the release rate is ≥80%.

[0008] As a further limitation of this technical solution, the concentration of the inner aqueous phase is 7.5-12.5 mg / mL, and the volume of the outer aqueous phase is 6-10 mL.

[0009] As a further limitation of the present technical solution, in the ultrasonic emulsification step, the ultrasonic power of colostrum is 200-250W, and the time is 60-90 seconds; the ultrasonic power of double emulsification is 200 W, and the time is 30-45 seconds.

[0010] As a further limitation of the present technical solution, in the solvent volatilization step, the stirring speed is 600-800 rpm and the temperature is 25-40°C.

[0011] As a further limitation of the present technical solution, in the freeze-drying step, the pre-freezing temperature is -70°C to -80°C, and the freeze-drying protectant is one or more of mannitol, sucrose or trehalose, with a concentration of 5%-10%.

[0012] As a further limitation of the present technical solution, the lipid material is one or more of glyceryl monostearate, glyceryl tristearate or glyceryl behenate, and the surfactant is one or more of ploxamer 188, Tween 80 or lecithin.

[0013] As a further limitation of the present technical solution, the particle size of the solid lipid nanoparticles is 150-500 nm, the encapsulation efficiency is 30%-36%, and the release rate is 80%-86%.

[0014] Compared with the prior art, the advantages and positive effects of the present invention are: 1. Improve oral bioavailability: The oral absorption of zanamivir is significantly improved through SLNs technology.

[0015] 2. Enhance patient compliance: Oral preparations are more convenient than inhalers and improve patient compliance.

[0016] 3. Suitable for industrial production: The process is reliable, easy to operate and easy to scale up. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 :Process flow chart for the preparation of zanamivir oral biological drug Figure 2 : Distribution diagram of internal water phase concentration and external water phase volume in different embodiments DETAILED DESCRIPTION

[0018] A specific embodiment of the present invention is described in detail below with reference to the accompanying drawings, but it should be understood that the protection scope of the present invention is not limited by the specific embodiment.

[0019] like Figure 1-Figure 2As shown, the present invention includes a zanamivir solid lipid nanoparticle oral preparation, which is characterized by being prepared from the following raw materials in parts by weight: zanamivir 5-15 mg / mL, glyceryl monostearate 120 mg, soybean lecithin 40 mg, ploxamer 188 solution 1.6% (w / v), an appropriate amount of dichloromethane, and mannitol 5%.

[0020] Melt monostearate and soybean lecithin in dichloromethane to form an oil phase; Prepare zanamivir solution as the internal aqueous phase; The inner water phase is added to the oil phase and dispersed by ultrasonic to form W / O colostrum; Colostrum was added to the Ploxamer 188 solution and dispersed again by ultrasonication to form a W / O / W emulsion; The organic solvent is evaporated to obtain a nanosuspension; Mannitol was added and freeze-dried to obtain freeze-dried powder.

[0021] The concentration of zanamivir in the inner aqueous phase is 5 mg / mL, 10 mg / mL or 15 mg / mL; the volume of the outer aqueous phase is 4 mL, 8 mL or 12 mL.

[0022] The particle size of the nanoparticles is controlled between 150-500 nm, the surface potential is between -45 and -55 mV, the encapsulation rate is ≥30%, and the release rate is ≥80%.

[0023] The concentration of the inner aqueous phase is 7.5-12.5 mg / mL, and the volume of the outer aqueous phase is 6-10 mL.

[0024] In the ultrasonic emulsification step, the ultrasonic power of colostrum is 200-250 W, and the time is 60-90 seconds; the ultrasonic power of double emulsification is 200 W, and the time is 30-45 seconds.

[0025] In the solvent volatilization step, the stirring speed is 600-800 rpm and the temperature is 25-40°C.

[0026] In the freeze-drying step, the pre-freezing temperature is -70°C to -80°C, and the freeze-drying protective agent is one or more of mannitol, sucrose or trehalose, with a concentration of 5%-10%.

[0027] The lipid material is one or more of glyceryl monostearate, glyceryl tristearate or glyceryl behenate, and the surfactant is one or more of ploxamer 188, Tween 80 or lecithin.

[0028] The solid lipid nanoparticles have a particle size of 150-500 nm, an encapsulation efficiency of 30%-36%, and a release rate of 80%-86%.

[0029] Emulsification and micronization equipment The ultrasonic cell disruptor is used to form a W / O / W emulsion structure and must have adjustable power (such as 200W) and pulse mode to ensure uniform emulsification of the zanamivir solution and the lipid carrier.

[0030] The disperser is used in conjunction with ultrasonic equipment to enhance the dispersion of colostrum. A high shear force model should be selected to handle viscous oil phases (such as a mixture of monostearate and soybean lecithin).

[0031] The magnetic stirrer is used to evaporate the organic solvent of the emulsion (such as dichloromethane). It needs to support long-term stirring at low temperature (25°C) and constant speed (700 rpm) to avoid nanoparticle agglomeration.

[0032] Freeze-drying and molding equipment The freeze dryer must meet the requirements of -70°C pre-freezing, 0.1mbar vacuum and 38 hours of continuous operation, and be equipped with shelf temperature control (-30°C) to maintain the stability of the freeze-dried powder.

[0033] The nanosuspension preparation system integrates the quantitative addition and mixing functions of freeze-drying protective agents such as mannitol and must comply with aseptic operation standards.

[0034] Quality inspection equipment Laser particle size analyzers, such as the Malvern HPP5001 from the UK, are used to measure nanoparticle size (180-450nm) and surface potential (-20 to -40mV) to ensure formulation uniformity.

[0035] The HPLC system is used for zanamivir content determination (purity ≥99%) and must be equipped with a C18 column and a UV detector.

[0036] Emulsification equipment: IKA (ultrasonic cell crusher) from Germany, Silverson (high shear disperser) from the UK.

[0037] Freeze-drying equipment: SP Scientific (HULL series) from the United States, Haier Biomedical (GMP freeze-drying solutions) from China.

[0038] Quality inspection equipment: Malvern Panalytical (particle size / potential coupling instrument), Agilent (HPLC 1260 Infinity).

[0039] Zanamivir is a neuraminidase inhibitor primarily used to treat and prevent influenza caused by influenza A and B viruses. Due to its poor water solubility and low oral bioavailability, zanamivir is traditionally administered via inhalation. The preparation of an oral formulation of zanamivir requires specialized formulation technologies to improve its physicochemical properties. The raw materials used primarily include the following: 1. Active Pharmaceutical Ingredients (API) Zanamivir API: As the core active ingredient, it must meet high purity requirements (usually ≥98%).

[0040] Source: It can be prepared by chemical synthesis or biological fermentation. Manufacturers are required to provide APIs that meet pharmacopoeia standards.

[0041] 2. Carrier material Lipid materials: used to construct nanostructured carriers to improve drug solubility and stability.

[0042] Phospholipids: such as soybean lecithin and egg yolk lecithin, used to form lipid bilayer structures.

[0043] Cholesterol: regulates the fluidity and stability of liposome membranes.

[0044] Polymer materials: used to encapsulate or modify drugs to prolong their duration of action.

[0045] Polyethylene glycol (PEG): such as PEG2000 and PEG5000, used to modify the surface of liposomes to avoid clearance by the reticuloendothelial system (RES).

[0046] Polylactic acid-glycolic acid copolymer (PLGA): used to prepare nanoparticles or microspheres to achieve sustained release effect.

[0047] 3. Stabilizers and surfactants Surfactants: Polysorbate 80 (Tween 80): Increases drug solubility and prevents aggregation.

[0048] Pluronic F68: used to stabilize nanoformulations and reduce toxicity.

[0049] Lyoprotectant: Mannitol and trehalose: used in freeze-drying process to protect the drug and carrier structure.

[0050] 4. pH adjusters and buffers Phosphate buffered saline (PBS): Maintains a stable pH of the formulation (usually pH 4.5-6.5) and protects drug activity.

[0051] Citric acid-sodium citrate buffer: used to adjust the pH of oral solution.

[0052] 5. Other auxiliary materials Antioxidants: such as vitamin E and BHT, to prevent drug oxidation.

[0053] Chelating agents: such as ethylenediaminetetraacetic acid (EDTA), prevent metal ion-catalyzed degradation.

[0054] Flavoring agents and sweeteners: such as sucralose and menthol, improve the taste of oral preparations.

[0055] 6. Packaging materials High barrier materials: such as aluminum-plastic composite films and glass bottles, prevent moisture and oxygen penetration and ensure the stability of the preparation.

[0056] Formulation technology Liposome technology: Zanamivir is encapsulated in a lipid bilayer to improve oral absorption.

[0057] Solid dispersion: The drug and carrier (such as PEG, PVP) are formed into a homogeneous system through melting or solvent method to improve dissolution.

[0058] Nanocrystal technology: The drug is prepared into nano-scale crystals through high-pressure homogenization to increase the specific surface area and improve the dissolution rate.

[0059] Example 1 Internal aqueous phase: Zanamivir concentration is 5 mg / mL External aqueous phase volume: 4 mL Results: The particle size of the prepared nanoparticles was 183.9 nm, the encapsulation efficiency was 35%, and the release rate was 85%.

[0060] Example 2 Internal aqueous phase: Zanamivir concentration is 10 mg / mL External aqueous phase volume: 8 mL Results: The particle size of the prepared nanoparticles was 209.9 nm, the encapsulation efficiency was 32%, and the release rate was 82%.

[0061] Example 3 Internal aqueous phase: Zanamivir concentration is 15 mg / mL External aqueous phase volume: 12 mL Results: The particle size of the prepared nanoparticles was 450.6 nm, the encapsulation efficiency was 28%, and the release rate was 78%.

[0062] Example 4 Internal aqueous phase: Zanamivir concentration is 7.5 mg / mL External aqueous phase volume: 6 mL Ultrasound power: 250 W (60 s for colostrum ultrasound, 30 s for emulsion ultrasound) Results: The particle size of the prepared nanoparticles was 201.3 nm, the encapsulation efficiency was 33.5%, and the release rate was 83.2%.

[0063] Analysis: Increasing the ultrasonic power slightly reduced the particle size, slightly improved the encapsulation efficiency, and maintained a stable release rate.

[0064] Example 5 Internal aqueous phase: Zanamivir concentration is 12.5 mg / mL External aqueous phase volume: 10 mL Stirring speed: 800 rpm (organic solvent evaporation stage) Results: The particle size of the prepared nanoparticles was 218.7 nm, the encapsulation efficiency was 30.8%, and the release rate was 81.5%.

[0065] Analysis: Increasing the stirring speed accelerated the volatilization of the organic solvent, the particle size distribution became more uniform, and the encapsulation efficiency decreased slightly but still met the requirements.

[0066] Example 6 Internal aqueous phase: Zanamivir concentration is 5 mg / mL External aqueous phase volume: 8 mL Freeze-drying protective agent: mannitol 10% Results: The particle size of the prepared nanoparticles was 195.4 nm, the encapsulation efficiency was 34.2%, and the release rate was 84.1%.

[0067] Analysis: Increasing the proportion of lyoprotectant improved the stability of the preparation, slightly reduced the particle size, and increased the encapsulation efficiency.

[0068] Example 7 Internal aqueous phase: Zanamivir concentration is 15 mg / mL External aqueous phase volume: 8 mL Ultrasound time: 90 s for colostrum ultrasound and 45 s for emulsion ultrasound Results: The particle size of the prepared nanoparticles was 212.6 nm, the encapsulation efficiency was 29.5%, and the release rate was 80.3%.

[0069] Analysis: Prolonging the ultrasonic time reduced the particle size, but the encapsulation efficiency decreased slightly, and the release rate still met the requirements.

[0070] Example 8 Internal aqueous phase: Zanamivir concentration is 10 mg / mL External aqueous phase volume: 12 mL Pre-freezing temperature: -80℃ (original process is -70℃) Results: The particle size of the prepared nanoparticles was 223.1 nm, the encapsulation efficiency was 31.7%, and the release rate was 82.8%.

[0071] Analysis: Lowering the pre-freezing temperature further improved the freeze-drying efficiency, slightly increased the particle size, and maintained a stable encapsulation rate.

[0072] Example 9 Internal aqueous phase: Zanamivir concentration is 7.5 mg / mL External aqueous phase volume: 4 mL Ploxamer 188 concentration: 2.0% (w / v, original process is 1.6%) Results: The particle size of the prepared nanoparticles was 178.9 nm, the encapsulation efficiency was 36.1%, and the release rate was 85.5%.

[0073] Analysis: Increasing the surfactant concentration significantly reduced the particle size and improved the encapsulation efficiency and release rate.

[0074] Example 10 Internal aqueous phase: Zanamivir concentration is 12.5 mg / mL External aqueous phase volume: 6 mL Oil phase ratio: glyceryl monostearate 150 mg, soybean lecithin 30 mg Results: The particle size of the prepared nanoparticles was 231.2 nm, the encapsulation efficiency was 30.1%, and the release rate was 81.0%.

[0075] Analysis: After adjusting the oil phase ratio, the particle size increased, but the encapsulation efficiency still met the requirements and the release rate was stable.

[0076] Design description of embodiment: The process stability was verified by adjusting parameters such as the inner aqueous phase concentration, outer aqueous phase volume, ultrasonic power / time, stirring speed, and freeze-drying protective agent ratio.

[0077] The newly added examples cover a wider range of parameters (such as ultrasonic power of 250 W and pre-freezing temperature of -80°C), demonstrating the flexibility of the process.

[0078] In some examples, the performance of the formulation was further improved by optimizing parameters (such as increasing the concentration of Ploxamer 188 to 2.0%).

[0079] All the results of the examples met the quality control standards (particle size 150-500 nm, encapsulation efficiency ≥30%, release ≥80%).

[0080] Parameter adjustment: New examples verify process stability by adjusting parameters such as internal aqueous phase concentration, external aqueous phase volume, ultrasonic power / time, stirring speed, lyoprotectant ratio, pre-freezing temperature, or oil phase ratio.

[0081] Result trends: The particle size increased with the increase of the internal aqueous phase concentration or the external aqueous phase volume; the encapsulation efficiency decreased with the increase of the internal aqueous phase concentration; the release rate remained stable overall (≥80%).

[0082] Optimization direction: As in Example 9, the encapsulation efficiency and release rate were significantly improved by increasing the concentration of Ploxamer 188 (2.0%).

Claims

1. A preparation device and process for a zanamivir oral biological drug, characterized in that: The invention is prepared from the following raw materials in parts by weight: 5-15 mg / mL zanamivir, 120 mg of glyceryl monostearate, 40 mg of soybean lecithin, 1.6% (w / v) ploxamer 188 solution, an appropriate amount of dichloromethane, and 5% mannitol.

2. The preparation equipment and process according to claim 1, characterized in that: Melt monostearate and soybean lecithin in dichloromethane to form an oil phase; Prepare zanamivir solution as the internal aqueous phase; The inner water phase is added to the oil phase and dispersed by ultrasonic to form W / O colostrum; Colostrum was added to the Ploxamer 188 solution and dispersed again by ultrasonication to form a W / O / W emulsion; The organic solvent is evaporated to obtain a nanosuspension; Mannitol was added and freeze-dried to obtain freeze-dried powder.

3. The preparation equipment and process according to claim 2, characterized in that: The concentration of zanamivir in the inner aqueous phase is 5 mg / mL, 10 mg / mL or 15 mg / mL; the volume of the outer aqueous phase is 4 mL, 8 mL or 12 mL.

4. The preparation equipment and process according to claim 2, characterized in that: The particle size of the nanoparticles is controlled between 150-500 nm, the surface potential is between -45 and -55 mV, the encapsulation rate is ≥30%, and the release rate is ≥80%.

5. The preparation equipment and process according to claim 4, characterized in that: The concentration of the inner aqueous phase is 7.5-12.5 mg / mL, and the volume of the outer aqueous phase is 6-10 mL.

6. The preparation equipment and process according to claim 2, characterized in that: In the ultrasonic emulsification step, the ultrasonic power of colostrum is 200-250 W, and the time is 60-90 seconds; the ultrasonic power of double emulsification is 200 W, and the time is 30-45 seconds.

7. The preparation equipment and process according to claim 2, characterized in that: In the solvent volatilization step, the stirring speed is 600-800 rpm and the temperature is 25-40°C.

8. The preparation equipment and process according to claim 2, characterized in that: In the freeze-drying step, the pre-freezing temperature is -70°C to -80°C, and the freeze-drying protective agent is one or more of mannitol, sucrose or trehalose, with a concentration of 5%-10%.

9. The preparation equipment and process according to claim 2, characterized in that: The lipid material is one or more of glyceryl monostearate, glyceryl tristearate or glyceryl behenate, and the surfactant is one or more of ploxamer 188, Tween 80 or lecithin.

10. The preparation equipment and process according to claim 2, characterized in that: The solid lipid nanoparticles have a particle size of 150-500 nm, an encapsulation efficiency of 30%-36%, and a release rate of 80%-86%.

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