Stiripentol nanosuspension, nanomicelles, and preparation method and application thereof

Abstract

The present invention provides a stiripentol nanosuspension, a nano-micelle, and a preparation method and application thereof. The stiripentol nano-suspension is prepared by taking a stabilizer, dissolving it in water by ultrasonication, and obtaining an aqueous phase; taking a stiripentol raw material drug, dissolving it in anhydrous ethanol, and obtaining an organic phase; injecting the organic phase into the aqueous phase at a preset speed according to a preset ratio at a certain stirring speed, stirring evenly, and obtaining a stiripentol nano-coarse suspension, and then high-pressure homogenization for multiple times, standing, and volatilizing the organic phase to obtain a stiripentol nano-micelle suspension; the stiripentol nano-micelles are prepared by adding a freeze-drying protective agent to the stiripentol nano-coarse suspension, and then high-pressure homogenization for multiple times, standing, and volatilizing the organic phase to obtain a stiripentol nano-micelle suspension; and solidifying by freeze-drying process to obtain stiripentol nano-micelles. The nano-micelles have good redispersibility and good stability, and the average particle size after redissolution is 100-300nm, and the dispersion coefficient PDI does not exceed 0.5.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to stiripentol nanosuspension, nanomicelles, and preparation methods and applications thereof. Background Art

[0002] Epilepsy is caused by abnormal high-frequency discharges of local neurons in the brain due to various reasons and spread to the surrounding normal brain tissue, resulting in abnormal brain function behaviors such as movement, sensation, consciousness or spirit, and presents as chronic neurological symptoms with intermittent and unpredictable attacks. Most of the patients are adolescents, and the onset is characterized by suddenness and persistence. If the onset cannot be prevented and effectively controlled, it will have a great impact on the patients and the people around them. There are many ways to treat epilepsy, including lifestyle and dietary changes, drug therapy, medical devices and surgical intervention. At present, the use of anti-epileptic drugs is still the first choice for epilepsy patients. About 30% of patients' conditions are not satisfactorily controlled. In addition, many patients have significant adverse reactions. Therefore, there is still unmet clinical demand in the anti-epileptic drug market, including refractory epilepsy and rare specific epilepsy. Therefore, it is urgent to find a drug that has good efficacy against epilepsy and few side effects.

[0003] Stiripentol (Molecular formula C 14 H 18 O3; molecular weight: 234.29; chemical name: 1-(benzo[D][1,3]dioxol-5-yl-4,4-dimethylpent-1-en-3-ol). It was developed by Biocodex in France and later authorized to Meiji Seika Pharmaceutical Co., Ltd. of Japan for joint development. In 2018, the US FDA approved the marketing of stiripentol capsules and stiripentol powder for oral suspension, with the trade name Diacomit. Biocodex is responsible for sales in Europe and the United States, and Meiji Seika Pharmaceutical Co., Ltd. is responsible for sales in Japan. Antiepileptic drugs are the main method for clinical epilepsy treatment. Since its launch, the clinical response has been good. Its mechanism of action is to inhibit cytochrome P450. It has a good therapeutic effect on patients with severe myoclonic epilepsy. In addition, the drug can reduce the side effects of antiepileptic drugs used in combination with it. Long-term use has little effect on patients and is an ideal drug for the treatment of epilepsy.

[0004] However, stiripentol (STP) is a BCS II drug with low water solubility and high permeability. It dissolves slowly and incompletely in the gastrointestinal tract and has poor stability in an acidic environment. Due to the above-mentioned limitations of oral absorption, the daily oral dose of the drug is relatively high. At present, the preparations available in the market at home and abroad include capsules 250 mg / 500 mg; oral dry suspension 250 mg / 500 mg, the site of action is the γGABA-α3 subunit, the dosage is 50 mg / kg / day, 2 to 3 times a day, the recommended maximum total dose is 3000 mg / day, and the blood concentration reaches the peak (4 to 22 μg / mL) 2 to 3 hours after a single dose, and it is generally necessary to take it with meals to achieve the best effect. This seriously limits its further clinical application, so it is necessary to adopt formulation technology to increase its solubility, improve its dissolution rate, promote its absorption in the body, and prepare stiripentol preparations with fast dissolution, high bioavailability and good stability.

[0005] Studies have shown that when poorly soluble drugs are made into nanosuspensions, their particle size reaches the nanometer level. The drug particles are suspended in the solution by relying on the charge effect or (and) steric effect of the surfactant, which can increase their dissolution rate and solubility, thereby improving the bioavailability of the preparation, reducing the dosage of the drug, and reducing toxic and side effects. At present, there are two methods for preparing nanosuspensions: bottom up and top down. The former includes precipitation, supercritical fluid technology, emulsification, etc., and the latter includes media grinding and high-pressure homogenization. In addition, the two technologies can be used in combination. At the same time, it should be noted that nanosuspensions, as highly dispersed nanocolloid systems, have high system free energy and are prone to stability problems such as aggregation, sedimentation, and maturation in aqueous media, which has become an important factor limiting their application.

[0006] So far, there is no nano-scale stiripentol preparation on the market at home and abroad. Therefore, designing a safe, effective and easy-to-use new stiripentol preparation has important clinical value. Summary of the invention

[0007] The purpose of the present invention is to provide a stiripentol nanosuspension, a nanomicelle, and a preparation method and application thereof in view of the above-mentioned deficiencies in the prior art.

[0008] To achieve the above object, the present invention adopts the following technical solution:

[0009] The first object of the present invention is to provide a stiripentol nanosuspension, which consists of stiripentol, a stabilizer and water for injection, wherein the mass ratio of stiripentol to the stabilizer is 1:(0.3-1.2), the concentration of stiripentol in the stiripentol nanosuspension is 1 mg / mL-5 mg / mL, and the stabilizer is any one of a nonionic surfactant, an anionic surfactant and a cationic surfactant.

[0010] Furthermore, the nonionic surfactant is any one of Tween 20, Tween 60 and Tween 80, the anionic surfactant is any one of sodium dodecyl sulfonate and sodium dodecyl sulfate, and the cationic surfactant is hexadecyltrimethylammonium bromide.

[0011] Furthermore, the particle size of the stiripentol particles in the stiripentol nanosuspension is 100 nm to 300 nm.

[0012] Furthermore, the stabilizer is sodium dodecyl sulfonate or sodium dodecyl sulfate, and the particle size of the stiripentol particles in the stiripentol nanosuspension is 100nm~200nm.

[0013] The second object of the present invention is to provide a method for preparing a stiripentol nanosuspension, comprising the following specific steps:

[0014] Step S1, preparation of stiripentol nanocoarse suspension

[0015] A stabilizer is measured and dissolved in water by ultrasound to obtain an aqueous phase; a stiripentol raw material is measured and dissolved in an organic solvent to obtain an organic phase; at a certain stirring speed, the organic phase is injected into the aqueous phase at a preset speed according to a preset ratio, and stirred evenly to obtain a stiripentol nanocoarse suspension;

[0016] Step S2, preparation of stiripentol nanosuspension

[0017] The crude suspension obtained in step S1 is subjected to high pressure homogenization for multiple times, and then allowed to stand to volatilize the organic phase, thereby obtaining a stiripentol nanomicelle suspension.

[0018] Furthermore, in step S1, the ultrasonic dissolution power of the stabilizer is 200w~450w, the volume ratio of the organic phase to the aqueous phase is 1:5~1:20, the organic phase injection speed is 2~10 mL / min, the stirring speed is 200~1000 r / min, and in step S2, the power of the high-pressure homogenization is 800~1200bar, and the homogenization cycle number of the high-pressure homogenization ranges from 6 to 30 cycles.

[0019] Furthermore, the organic solvent is one or a combination of methanol, ethanol, acetone, isopropanol, acetonitrile, DMSO, DMF and ethyl acetate.

[0020] The third object of the present invention is to provide a stiripentol nano micelle, which is prepared by adding a lyophilization protective agent to the above stiripentol nano coarse suspension, homogenizing under high pressure, and freeze-drying and solidifying.

[0021] Furthermore, the lyophilization protective agent is any one of sucrose, mannose and lactose, and the mass concentration of the lyophilization protective agent is 2%~10%; the power of high-pressure homogenization is 800~1200bar, and the homogenization cycle number of high-pressure homogenization ranges from 6 to 30 weeks.

[0022] The fourth object of the present invention is to provide an anti-epileptic drug composition, which is prepared by using the above-mentioned stiripentol nanosuspension, or prepared by using the above-mentioned stiripentol nanomicelles.

[0023] Antiepileptic drugs refer to compositions comprising stiripentol nanosuspensions or stiripentol nanomicelles and at least one selected from the following pharmaceutically acceptable and pharmacologically compatible components: fillers, solvents, diluents, carriers, excipients, distributing agents and acceptors, delivery agents such as preservatives, stabilizers, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, thickeners, sweeteners, flavoring agents, aromatics, antibacterial agents, fungicides, lubricants and extended delivery control agents, the selection and proportion of which depend on the nature and route of administration and the dosage. Examples of suitable suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ethers, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth gum and mixtures thereof. Various antibacterial and antifungal agents such as parabens, chlorobutanol, sorbic acid, etc. can be used to provide protection against microorganisms. Antiepileptic drugs may also include isotonic agents such as sugars, sodium chloride, etc. The sustained action of the composition can be achieved by using agents that slow down the absorption of the active ingredient, such as aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery agents include water, ethanol, polyols and mixtures thereof, natural oils (such as olive oil) and organic esters (such as ethyl oleate) for injection. Examples of fillers are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate, etc. Examples of disintegrants and dispensing agents are starch, alginic acid and its salts and silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talcum and high molecular weight polyethylene glycol. Pharmaceutical compositions for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous and topical or rectal administration of active ingredients alone or in combination with another active compound can be used as a mixture with traditional pharmaceutical carriers to be applied to animals and humans in standard administration forms. Suitable standard administration forms include oral forms, such as tablets, capsules, pills, powders, granules, chewing gum and oral solutions or suspensions; sublingual and buccal administration forms; aerosols; implants; topical, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular forms; and rectal administration forms.

[0024] Compared with the prior art, the technical solution provided by the present invention has the following beneficial effects:

[0025] (1) The stiripentol nanosuspension prepared by the present invention is a nanoscale drug particle made of stiripentol, which has a small particle size, a narrow particle size distribution, good stability, and is not easy to precipitate; a small amount of safe, injectable stabilizer is added to overcome the insolubility problem of stiripentol, improve the solubility of the drug, increase the accumulation of stiripentol in the target area, reduce its toxic and side effects on normal tissues, and improve the in vivo distribution of the drug; reduce the dosage of the drug; improve the safety of the drug and increase the compliance of the subjects; improve the bioavailability of the drug in the body; reduce the adverse reactions and toxic and side effects of the drug. It has a better therapeutic effect and has a wider applicability. The stiripentol nanosuspension prepared by the present invention has a simple prescription process, a surfactant as a pharmaceutical excipient, a small amount, high safety, no irritation, no physiological toxicity, good biocompatibility, low cost, and easy industrialization.

[0026] (2) By freeze-drying the stiripentol nanosuspension into a solid powder, the particle agglomeration and particle size increase caused by long-term storage of the suspension can be avoided, and the stiripentol nanomicelles have good redispersibility and stability. The average particle size after reconstitution is 100-300nm, and the dispersion coefficient PDI does not exceed 0.5. It has the characteristics of fast dissolution rate, good stability, and high drug content.

[0027] (3) The pharmaceutical composition containing stiripentol nanosuspension or stiripentol nanomicelles of the present invention can be used to treat and / or prevent epilepsy. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The figure is a comparison of the appearance of stiripentol nanosuspensions prepared using different stabilizers;

[0029] Figure 2 is the XRD pattern of stiripentol nanomicelles;

[0030] Figure 3 This is the in vitro dissolution curve of stiripentol nanomicelles, stiripentol physical mixture, and stiripentol raw material in water. DETAILED DESCRIPTION

[0031] If no specific technology or conditions are specified in the present invention, the technology or conditions described in the literature in the field or the product instructions shall be followed. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0032] The reagent information used in this implementation is as follows

[0033] Tween 20, abbreviated as T20, CAS number: 9005-64-5; Tween 60, abbreviated as T60, CAS number: 9005-67-8; sodium dodecyl sulfonate, CAS number: 2386-53-0; sodium dodecyl sulfate, abbreviated as SDS, CAS number: 151-21-3; hexadecyltrimethylammonium bromide, abbreviated as CTAB, CAS number: 57-09-0; poloxamer 188, abbreviated as F68, CAS number: 9003-11-6, poloxamer 407, abbreviated as F127, CAS number: 9003-11-6.

[0034] The stiripentol dry suspension used in the present invention is provided by Biocodex Company and is commercially available.

[0035] The performance characterization method of stiripentol nanosuspension and nanomicelles is as follows:

[0036] 1) Determination of Size, PDI and Zeta

[0037] The size, PDI and Zeta potential of the STP solution were measured using a Malvern particle size analyzer. Experimental conditions: solute is STP, refractive index is 1.578, solvent is water, refractive index is 1.330, temperature is 25°C, equilibrium time is 60 s, parallel measurement is performed 3 times, and the average value is calculated.

[0038] 2) X-ray diffraction (PXRD)

[0039] The STP nanomicelle powder, SDS powder and the powder of the physical mixture were evenly spread in the sample tank, and the surface was pressed flat with a glass slide, and the sample holder was fixed on the PXRD for testing. The measurement conditions were as follows: the anode target was a Cu-Kα target, the tube voltage was 40 kV, the tube current was 40 mA, the scanning speed was 0.1 s / step, the scanning step was 0.02° per step, and the scanning range was from 4° to 50° (2θ).

[0040] In order to make the objectives, technical solutions and advantages of the present invention more clear, the specific implementation methods of the present invention are further described in detail below in conjunction with specific embodiments and drawings.

[0041] STP: stiripentol.

[0042] PM: stiripentol physical mixture.

[0043] The instruments and equipment involved in the examples are described as follows: freeze dryer (FreeZone) from Labconco, USA, and high pressure homogenizer (AH-NANO) from ATS Industrial Systems Co., Ltd.

[0044] The invention adopts a reverse precipitation method combined with a high-pressure homogenization method to prepare a stiripentol nanosuspension, adopts freeze drying to solidify the stiripentol nanosuspension to prepare stiripentol nanomicelles, and takes the average particle size of the micelles and PDI as evaluation indicators.

[0045] Example 1

[0046] Preparation of stiripentol nanosuspension

[0047] Weigh an appropriate amount of stabilizer T20, dissolve them in water under the condition of ultrasonic power of 250w, and obtain an aqueous phase with a concentration of 2.5mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6 mL / min at a stirring speed of 1000 r / min according to the volume ratio of organic phase to aqueous phase of 1:10, to obtain a coarse suspension with a STP drug loading of 4mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and under the condition of a homogenization power of 1000bar in the high-pressure homogenizer, after 10 weeks of high-pressure homogenization, place it at room temperature for 24h, volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) are measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 200.4nm, a polydispersity index of 0.257, and a potential of -8.19mV. And the particle size has basically not changed after being placed for 24 hours.

[0048] Example 2

[0049] Preparation of stiripentol nanosuspension

[0050] Weigh an appropriate amount of stabilizer T60, dissolve them in water under the condition of ultrasonic power of 250w, and obtain an aqueous phase with a concentration of 2.5mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6 mL / min at a stirring speed of 1000 r / min according to the volume ratio of organic phase to aqueous phase of 1:10, to obtain a coarse suspension with a STP drug loading of 4mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and under the condition of a homogenization power of 1000bar in the high-pressure homogenizer, after 10 weeks of high-pressure homogenization, place it at room temperature for 24h, volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) were measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 151.8nm, a polydispersity index of 0.097, and a potential of -6.23mV. And the particle size has basically not changed after being placed for 24 hours.

[0051] Example 3

[0052] Preparation of stiripentol nanosuspension

[0053] Weigh an appropriate amount of stabilizer CTAB, dissolve them in water under the condition of ultrasonic power of 250w, and obtain an aqueous phase with a concentration of 2.5mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6 mL / min at a stirring speed of 1000 r / min according to the volume ratio of organic phase to aqueous phase of 1:10 to obtain a coarse suspension with a STP drug loading of 4mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24h after 6 weeks of high-pressure homogenization under the condition of a homogenization power of 1000bar, volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) are measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 287.6nm, a polydispersity index of 0.098, and a potential of -6.35mV. And the particle size has basically not changed after being placed for 24 hours.

[0054] Example 4

[0055] Preparation of stiripentol nanosuspension in this embodiment

[0056] Weigh an appropriate amount of stabilizer SDS, dissolve it in water under the condition of ultrasonic power of 200w, and obtain an aqueous phase with a concentration of 1.5mg / mL. Weigh an appropriate amount of STP raw material, dissolve it in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 2mL / min at a stirring speed of 200 r / min according to the volume ratio of organic phase to aqueous phase of 1:7.5 to obtain a coarse suspension with a STP drug loading of 3mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24h after 14 weeks of high-pressure homogenization under the condition of a homogenization power of 800bar, volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) were measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 103.5nm and a polydispersity index of 0.257. And the particle size has basically not changed after being placed for 24 hours.

[0057] Example 5

[0058] Preparation of stiripentol nanosuspension

[0059] Weigh an appropriate amount of stabilizer SDS, dissolve them in water under the condition of ultrasonic power of 350w, and obtain an aqueous phase with a concentration of 2.0mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6mL / min at a volume ratio of 1:15 between the organic phase and the aqueous phase under the condition of stirring speed of 600 r / min to obtain a coarse suspension with a STP drug loading of 4mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24h after 18 weeks of high-pressure homogenization under the condition of homogenization power of the high-pressure homogenizer of 1100bar to volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) were measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 130.3nm and a polydispersity index of 0.189. And the particle size has basically not changed after being placed for 24 hours.

[0060] Example 6

[0061] Preparation of stiripentol nanosuspension

[0062] Weigh an appropriate amount of stabilizer SDS, dissolve them in water under the condition of ultrasonic power of 450w, and obtain an aqueous phase with a concentration of 3.0mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 8mL / min at a volume ratio of 1:20 between the organic phase and the aqueous phase under the condition of stirring speed of 800 r / min to obtain a coarse suspension with a STP drug loading of 5mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24h after 30 cycles of high-pressure homogenization under the condition of homogenization power of the high-pressure homogenizer of 1200bar to volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) were measured by a laser particle size analyzer. The prepared stiripentol nanosuspension had an average particle size of 172.4nm and a polydispersity index of 0.352. And the particle size did not change substantially after being placed for 24 hours.

[0063] Comparative Example 1

[0064] Preparation of stiripentol nanosuspension

[0065] Weigh an appropriate amount of stabilizer poloxamer 188, dissolve them in water under ultrasonic power of 250w, and obtain an aqueous phase with a concentration of 2.5 mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6 mL / min at a stirring speed of 1000 r / min according to the volume ratio of organic phase to aqueous phase of 1:10 to obtain a coarse suspension with a STP drug loading of 4 mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24 hours after 10 weeks of high-pressure homogenization under the condition of a homogenization power of 1000bar, volatilize the organic phase, and obtain a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) are measured by a laser particle size analyzer. The prepared stiripentol nanosuspension has an average particle size of 2055nm and a polydispersity index of 1.002.

[0066] Comparative Example 2

[0067] Preparation of stiripentol nanosuspension

[0068] Weigh an appropriate amount of poloxamer 407, dissolve them in water under an ultrasonic power of 250w, and obtain an aqueous phase with a concentration of 2.5mg / mL. Weigh an appropriate amount of STP raw material, dissolve them in anhydrous ethanol, and inject the organic phase into the aqueous phase at a rate of 6 mL / min at a stirring speed of 1000 r / min according to the volume ratio of the organic phase to the aqueous phase of 1:10 to obtain a coarse suspension with a STP drug loading of 4mg / mL. Add the above coarse suspension to a high-pressure homogenizer (HPH), and place it at room temperature for 24h after 10 cycles of high-pressure homogenization under a homogenization power of 1000bar to volatilize the organic phase, thereby obtaining a stiripentol nanomicelle suspension. The particle size (Size) and dispersion index (PDI) were measured using a laser particle size analyzer. The prepared stiripentol nanosuspension had an average particle size of 1625nm and a polydispersity index of 0.970.

[0069] like Figure 1 As shown, it shows that: at the same concentration of the six stabilizers, F68 and F127 have precipitation and large particle size; the micelle particle size of T60 and CTAB is smaller, but they show a larger dispersion coefficient; the Size and PDI of SDS and T20 are not significantly different, but the smaller the particle size, the more violent the Brownian motion of the micelle in the system, and there will be agglomeration between particles, so electrostatic repulsion is needed to maintain the stability of the system. The potential of SDS (-40.73 mV) is greater than that of T20 (-8.19 mV), so SDS can be used as the best stabilizer for stiripentol nanomicelles.

[0070] Example 7

[0071] Preparation of stiripentol nanomicelles

[0072] The stiripentol nanocoarse suspension prepared in Example 6 was used to prepare stiripentol nanomicelles.

[0073] A freeze-drying protective agent 5% lactose was added to the above stiripentol nanocoarse suspension, and then the high-pressure homogenization pressure was 1200 bar. After 10 weeks of high-pressure homogenization, the organic phase was volatilized and the stiripentol nanomicelle suspension was obtained; the stiripentol nanomicelle suspension was pre-frozen at -80 ° C and then transferred to a freeze dryer for 24 h to obtain stiripentol nanomicelles. The particle size (Size) and dispersion index (PDI) were measured by a laser particle size analyzer, and the microscopic morphology of the stiripentol nanomicelles was observed by a scanning electron microscope.

[0074] As shown in Table 1, the biological particle size of the STP nanomicelles prepared in this example after re-dissolution is 232.0 nm, the PDI is 0.465, the Zeta is -57.13 mV, and the Zeta is negatively charged, indicating that the outermost layer of the micelle is the electron layer spontaneously formed by SDS.

[0075] Table 1. Particle size, PDI and Zeta results of stiripentol nanomicelles (n=3)

[0076]

[0077] like Figure 2 As shown, the blank material has an obvious diffraction peak, and both the STP physical mixture and the STP micelle lyophilized powder have diffraction peaks at this position, indicating that this is the diffraction peak of the material itself, and physical mixing does not affect the diffraction peak of the material. The STP micelle lyophilized powder and spectrum are similar to the blank material and do not show the characteristic peaks of STP, indicating that the STP micelles can be perfectly wrapped by the material.

[0078] Example 8

[0079] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the freeze-drying protective agent is 10% lactose.

[0080] Example 9

[0081] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the freeze-drying protective agent is 5% mannose.

[0082] Example 10

[0083] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the freeze-drying protective agent is 10% mannose.

[0084] Embodiment 11

[0085] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the freeze-drying protective agent is 5% sucrose.

[0086] Example 12

[0087] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the freeze-drying protective agent is 10% sucrose.

[0088] The microscopic morphology of the stiripentol nanomicelles prepared in Examples 7-12 after reconstitution was investigated. It was found that the nanomicelles with 5% lactose and 10% sucrose showed a better separation state under a scanning electron microscope after reconstitution. The average particle size after reconstitution was 100-300 nm, and the dispersion coefficient PDI did not exceed 0.5.

[0089] Comparative Example 3

[0090] The preparation of stiripentol nanomicelles in this example is basically the same as that in Example 7, except that the pre-freezing temperature is -20°C. The particle size (Size) and the dispersion index (PDI) are measured using a laser particle size analyzer. The suspension of the prepared stiripentol nanomicelles after reconstitution has an average particle size of 2253 nm and a polydispersity index of 0.835.

[0091] Comparative Example 4

[0092] The preparation of stiripentol nano-micelles in this example is basically the same as that in comparative example 3, except that the freeze-dried protective agent is 10% sucrose. The particle size (Size) and the dispersion index (PDI) are measured by a laser particle size analyzer. The suspension of the prepared stiripentol nano-micelles after reconstitution has an average particle size of 718.3 nm and a polydispersity index of 0.581.

[0093] Comparative Example 5

[0094] The freeze-drying protective agent was added by external addition method. The test content was as follows: 5% lactose, a freeze-drying protective agent, was added to the stiripentol nanosuspension prepared in Example 1, mixed well, and then pre-frozen at -80°C and then transferred to a freeze dryer for drying for 24 h to obtain stiripentol nanomicelles. The particle size (Size) and the dispersion index (PDI) were measured. The suspension of the obtained stiripentol nanomicelles after reconstitution had an average particle size of 1022 nm and a polydispersity index of 0.750.

[0095] Comparative Example 6

[0096] The preparation of stiripentol nanomicelles in this example is basically the same as that in comparative example 5, except that 10% sucrose is used as a freeze-dried protective agent. The particle size (Size) and the dispersion index (PDI) are measured. The suspension of the prepared stiripentol nanomicelles after reconstitution has an average particle size of 478.2 nm and a polydispersity index of 0.586.

[0097] In order to better explore the bioavailability characteristics of stiripentol nanomicelles, an in vitro dissolution study was conducted. The specific contents are as follows:

[0098] STP nanomicelles (NP-STP): stiripentol nanomicelles prepared in Example 7;

[0099] STP physical mixture (PM): stiripentol, T20 and lactose used in Example 7 are mixed uniformly to obtain a STP physical mixture (PM).

[0100] The dissolution curve of STP nanomicelles (NP-STP) was determined by the small cup method. The determination conditions were as follows: the dissolution medium was water, the volume of the dissolution medium was 250 mL, the rotation speed was 100 rpm / min, and the temperature was 37.0°C. Three portions of STP, physical mixture, and STP nanomicelle samples (equivalent to 10 mg of STP) were weighed respectively, accurately weighed, and put into the dissolution cup at the same time. The timing started when the API contacted the dissolution medium, and samples were taken at 5, 10, 20, 30, 45, 60, 90, and 120 min, 5 mL was drawn each time, and 5 mL of dissolution medium was replenished in time, filtered with a 0.45 μm pore size filter membrane, and the filtrate was taken as the test sample. The absorbance was measured at a wavelength of 270 nm by ultraviolet spectrophotometry.

[0101] The results are as follows Figure 3 As shown, compared with stiripentol raw materials, stiripentol nanomicelles have a faster in vitro release rate, and the cumulative in vitro dissolution in 30 minutes can reach more than 90%, achieving the effect of rapid dissolution.

[0102] In the absence of conflict, the above embodiments and features in the embodiments may be combined with each other.

[0103] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A stiripentol nanosuspension, characterized in that The stiripentol nanosuspension is composed of stiripentol, a stabilizer and water for injection, wherein the mass ratio of the stiripentol to the stabilizer is 1:(0.3-1.2), the concentration of stiripentol in the stiripentol nanosuspension is 1 mg / mL-5 mg / mL, and the stabilizer is sodium dodecyl sulfate; the stiripentol nanosuspension is prepared according to the following steps: Step S1, preparation of stiripentol nanocoarse suspension, A stabilizer is measured and dissolved in water by ultrasound to obtain an aqueous phase; a stiripentol raw material is measured and dissolved in an organic solvent to obtain an organic phase; at a certain stirring speed, the organic phase is injected into the aqueous phase at a preset speed according to a preset ratio, and stirred evenly to obtain a stiripentol nanocoarse suspension; The ultrasonic dissolution power of the stabilizer is 200w~450w, the volume ratio of the organic phase to the aqueous phase is 1:5~1:20, the organic phase injection speed is 2~10 mL / min, and the stirring speed is 200~1000 r / min; Step S2, preparation of stiripentol nanosuspension, The crude suspension obtained in step S1 is subjected to high-pressure homogenization for multiple times, and then allowed to stand to volatilize the organic phase, thereby obtaining a stiripentol nanomicelle suspension; the power of the high-pressure homogenization is 800-1500 bar, and the number of homogenization cycles of the high-pressure homogenization ranges from 6 to 30 cycles.

2. The stiripentol nanosuspension according to claim 1, wherein The particle size of the stiripentol particles in the stiripentol nanosuspension is 100-300 nm.

3. The stiripentol nanosuspension according to claim 2, characterized in that The particle size of the stiripentol particles in the stiripentol nanosuspension is 100nm-200nm.

4. The stiripentol nanosuspension according to claim 1, characterized in that The organic solvent is one or a combination of methanol, ethanol, propylene glycol, acetone, isopropanol, acetonitrile, DMSO, DMF and ethyl acetate.

5. A stiripentol nanomicelle, characterized in that: The stiripentol nanocoarse suspension according to any one of claims 1 to 4 is added with a freeze-drying protective agent, homogenized under high pressure, and freeze-dried and solidified to obtain the product.

6. A stiripentol nanomicelle according to claim 5, characterized in that: The lyoprotectant is any one of sucrose, mannose and lactose.

7. A stiripentol nanomicelle according to claim 6, characterized in that: The mass concentration of the lyophilization protectant is 2% to 10%.

8. A stiripentol nanomicelle according to claim 7, characterized in that: The power of high pressure homogenization is 800~1200bar.

9. A stiripentol nanomicelle according to claim 8, characterized in that: The homogenization cycles of high pressure homogenization range from 6 to 30 cycles.

10. An anti-epileptic drug composition, characterized in that: The stiripentol nanosuspension is prepared according to any one of claims 1 to 4, or the stiripentol nanomicelles are prepared according to any one of claims 5 to 9.

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