Lyophilized buagafuran series compound flashtab and method for preparing same

Abstract

Provided are a lyophilized buagafuran flashtab and a method for preparing same. The provided lyophilized buagafuran flashtab comprises the following raw materials in percentage by mass: 10%-90% of a buagafuran-mEV substance solution, 1%-5% of a binder, 0.1%-2% of a suspending agent, 0.1%-1% of a surfactant, 0.1%-1% of a sweetener, 0.1%-1% of a flavoring agent, and the balance of water. The surfactant is selected from polysorbate 80. The buagafuran is one or more of buagafuran and in-vivo metabolites thereof. Also provided is a method for preparing the lyophilized buagafuran flashtab. The lyophilized flashtab is prepared by lyophilization technology, thus possessing good stability, dissolution performance, and desirability for the oral absorption of buagafuran. Furthermore, the method features simplification and ease to implement.

Description

Freeze-dried orally disintegrating tablets of buguefuran series compounds and preparation method thereof

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on October 31, 2023, with application number 2023114364089 and invention name “Freeze-dried orally disintegrating tablets of buagafuran series compounds and preparation method thereof”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a freeze-dried orally disintegrating tablet of a buagafuran series compound and a preparation method thereof. Background Art

[0003] Agarwood is a traditional and precious Chinese medicinal herb. It is the resin-containing heartwood of the Aquilaria plant, a member of the Thymelaeaceae family, infected by fungi. The "Shennong Bencao Jing" (Classic of Materia Medica) lists agarwood as a top-grade medicinal herb. The "Compendium of Materia Medica" (Compendium of Materia Medica) states that agarwood has the effects of clearing the mind, invigorating qi, and harmonizing the spirit. Numerous reports have been published on the chemical composition of agarwood, primarily focusing on its essential oil. Chemists both domestically and internationally have reported on five classes of compounds: benzyl acetone, hydrocinnamic acid, linalools, agarwood spirocycles, and agarwood furans.

[0004] Buagafuran (BFN), chemically known as (1R,6S,9R)6,10,10-trimethyl-2-butyl-oxatricyclo[7.2.1.0 1,6 Dodeca-2-ene is a derivative of agarwood furan, a chemical component found in agarwood, and is prepared by chemical synthesis. Bugafuran has demonstrated anxiolytic effects in various animal studies with minimal toxic side effects. Bugafuran is an oily liquid at room temperature, highly lipophilic, poorly water-soluble, and unstable. Significant degradation and a decrease in content occur after three months at room temperature, necessitating long-term storage in a refrigerator (2-8°C). Traditional oral formulations of bugafuran suffer from low oral bioavailability and high clinical dosage requirements.

[0005] To make it into a solid oral preparation, the key issues that need to be solved include: solidifying the liquid drug to facilitate the design of solid oral preparations; improving drug stability to facilitate long-term storage of the preparation; and improving oral bioavailability to reduce the dosage.

[0006] Existing solid preparations of buagafuran are primarily capsules, containing dispersants and antioxidants, resulting in complex ingredients. Furthermore, the production process for these preparations requires multiple high-temperature steps, which is detrimental to maintaining the structure and content stability of the active ingredient and makes scale-up difficult. Existing buagafuran formulations also exhibit poor storage stability and are susceptible to oxidation when exposed to light or air.

[0007] Summary of the Invention

[0008] In view of this, the technical problem to be solved by the present invention is to provide a freeze-dried orally disintegrating tablet of a buagafuran series compound and a preparation method thereof. The prepared freeze-dried orally disintegrating tablet of a buagafuran series compound has high drug stability and dissolution efficiency, and is conducive to oral sublingual absorption.

[0009] On one hand, the present invention provides mEV-encapsulated bougainvulan compound nanoparticles, which are nanoparticles formed by using mEV as a carrier to encapsulate bougainvulan compounds.

[0010] The present invention adopts mEV as a carrier to encapsulate bougainvulan compounds.

[0011] Mesoporin (mEV) is a type of extracellular vesicle (ECV) (mEV), a type of biological nanoparticle formed from a phospholipid bilayer membrane that can be extracted in large quantities from milk. Its natural function includes delivering bioactive molecules (proteins, small nucleic acids, etc.) from the mother (cow) to the offspring (calf) through the gastrointestinal tract. mEVs are resistant to damage by digestive enzymes and low pH in the digestive tract, and are absorbed into the bloodstream through the digestive tract. Due to the widespread consumption of milk, long-term exposure to mEVs (through milk consumption) in most populations suggests that oral administration of mEVs is safe and tolerable in humans. Because mEVs are formed from a lipid bilayer membrane, their structure is ideal for encapsulating lipophilic small molecule drugs. Encapsulated buguafuran in mEVs isolates the active ingredient from air, effectively preventing oxidation, eliminating the need for antioxidants, and avoiding sublimation losses during the freeze-drying process. Furthermore, upon dissolution in the mouth, mEVs emit a natural milky aroma that effectively masks the bitter taste of the active drug. Therefore, mEV-encapsulated buguafuran drug formulations offer advantages over traditional solid dosage forms in terms of bioavailability, safety, and clinical compliance.

[0012] The present invention has no particular limitation on bugafuran compounds, including but not limited to bugafuran APIs and one or more of their metabolites in vivo.

[0013] The present invention provides a method for preparing the above-mentioned mEV-loaded bougainvulan compound nanoparticles, comprising the following steps:

[0014] The mEV particle solution and the bougainvulan compound solution are mixed, and then heated and ultrasonically treated to obtain mEV-encapsulated bougainvulan compound nanoparticles.

[0015] Optionally, the solvent in the solution of mEV particles is 1×PBS buffer.

[0016] Optionally, in the solution of the bugarfuran compound, the solvent is anhydrous ethanol.

[0017] The present invention does not particularly limit the mixing order of the mEV particle solution and the bugafuran compound solution. The mEV particle solution can be added to the bugafuran compound solution, or the bugafuran compound solution can be added to the mEV particle solution.

[0018] After the solution of the mEV particles is mixed with the solution of the buprenorphine compound, the final concentration of anhydrous ethanol is preferably no more than 30% v / v.

[0019] The heating temperature is preferably 60° C., and the heating time is preferably 5 h.

[0020] The power of the ultrasonic treatment is preferably 99% power, the temperature of the ultrasonic treatment is preferably 20° C., and the time of the ultrasonic treatment is preferably 18 hours.

[0021] The ratio of the bugrafuran compound to the mEV particles is preferably: 1×10 10 ~1×10 13 mEV particles.

[0022] The present invention uses the above-mentioned mEV-encapsulated bugafuran compound nanoparticles as raw materials to prepare freeze-dried orally disintegrating tablets, which can improve the stability and dissolution efficiency of bugafuran drugs and facilitate oral sublingual absorption.

[0023] The present invention also provides a bugrafuran freeze-dried orally disintegrating tablet comprising the following raw materials in the following mass percentages:

[0024] 10%-90% bugrafuran-mEV stock solution, 1%-5% binder, 0.1%-2% suspending agent, 0.1%-1% surfactant, 0.1%-1% sweetener, 0.1%-1% flavoring agent, and the balance is water;

[0025] The surfactant is selected from polysorbate 80.

[0026] Compared to other tablet or capsule dosage forms, mEV-encapsulated buagafuran in freeze-dried orally disintegrating tablets offers significant advantages. These tablets do not require water to be taken; saliva causes them to rapidly disintegrate or dissolve in the mouth. Buagafuran is rapidly metabolized by the liver, and a significant portion of the drug is absorbed orally after administration as a freeze-dried orally disintegrating tablet. This reduces the first-pass effect in the liver, effectively prolonging the drug's duration of action and increasing bioavailability. Furthermore, mEV encapsulation avoids the sublimation loss of low-melting-point compounds like buagafuran during the vacuum process, while also improving the formulation's flavor, making it more palatable for patients.

[0027] Furthermore, the mass content of the bucuronium-mEV stock solution is preferably 10% to 90%, more preferably 80% to 90%.

[0028] Furthermore, the mass content of the adhesive is preferably 1% to 5%, more preferably 3% to 5%.

[0029] Furthermore, the adhesive is selected from one of pullulan and gelatin; preferably, the adhesive is pullulan.

[0030] Furthermore, the mass content of the suspending agent is preferably 0.1% to 2%, more preferably 0.1% to 0.5%.

[0031] Furthermore, the suspending agent is selected from one of gum arabic, gum tragacanth, peach gum and xanthan gum; preferably, the suspending agent is xanthan gum.

[0032] Furthermore, the mass content of the surfactant is preferably 0.1% to 1%, more preferably 0.1% to 0.5%.

[0033] Furthermore, the mass content of the sweetener is preferably 0.1% to 1%, more preferably 0.1% to 0.5%.

[0034] Furthermore, the sweetener is selected from one of sucralose, sucrose, steviol glycoside, and saccharin sodium; preferably, the sweetener is sucralose.

[0035] Furthermore, the weight content of the flavoring agent is preferably 0.1% to 1%, more preferably 0.1% to 0.5%.

[0036] Furthermore, the flavoring agent is selected from one of sweet orange flavor, strawberry flavor, mEV, cherry flavor, and orange flavor; preferably, the flavoring agent is mEV.

[0037] Furthermore, the raw materials for preparing the bugrafuran-mEV stock solution are: 1×10 10 ~1×10 13 mEV particles.

[0038] In the present invention, the stability of bugrafuran after mEV nanoencapsulation is improved because most of the air is isolated. No antioxidant needs to be added when preparing freeze-dried orally disintegrating tablets, and the prepared drug can also obtain high stability.

[0039] In some specific embodiments of the present invention, the bugrafuran freeze-dried orally disintegrating tablets provided herein contain the following raw materials in percentage by weight: 82% bugrafuran-mEV stock solution, 4.6% binder, 0.2% suspending agent, 0.4% surfactant, 0.4% sweetener, 0.4% flavoring agent, and the balance water. In these embodiments, the raw materials are calculated based on their weight prior to lyophilization.

[0040] In some specific embodiments of the present invention, the bugrafuran freeze-dried orally disintegrating tablet comprises: bugrafuran-mEV stock solution, pullulan, xanthan gum, polysorbate 80, sucralose, mEV, and water.

[0041] In some specific embodiments of the present invention, the bugrafuran freeze-dried orally disintegrating tablets provided by the present invention contain the following raw materials in percentage by weight: 10%-90% bugrafuran-mEV stock solution, 1%-5% pullulan, 0.1%-2% xanthan gum, 0.1%-1% polysorbate 80, 0.1%-1% sucralose, 0.1%-1% mEV, and the balance is water.

[0042] The orally disintegrating tablets provided by the present invention utilize a well-chosen selection of excipients, including surfactants, such as binders or suspending agents, resulting in superior disintegration and sublingual absorption. Preliminary experiments have shown that, compared with other excipients, the combination of pullulan, xanthan gum, and polysorbate 80 is more effective in maintaining the stability of bugrafuran-mEV, improving oral release and sublingual absorption.

[0043] The present invention provides a method for preparing the above-mentioned bugrafuran freeze-dried orally disintegrating tablets, comprising the following steps:

[0044] (1) Loading buprenorphine onto mEVs to obtain buprenorphine-mEV stock solution;

[0045] (2) The buprenorphine-mEV stock solution was concentrated by ultrafiltration to obtain a concentrated solution;

[0046] (3) mixing the concentrated solution with a surfactant in a solution to obtain an inclusion solution; mixing the inclusion solution with a binder, a suspending agent, a sweetener, and a flavoring agent to obtain a drug solution;

[0047] (4) shearing the liquid medicine in step (3);

[0048] (5) degassing the liquid medicine after shearing in step (4);

[0049] (6) filling the degassed liquid medicine in step (5) into the cavity;

[0050] (7) Pre-freeze the filled liquid medicine;

[0051] (8) Freeze-dry the pre-frozen drug, setting the initial temperature to -30°C. The specific freeze-drying process is as follows:

[0052] Cold soaking temperature: ≤-60℃; Vacuum control: ≤200ubar

[0053] Plate temperature: from -30℃ to -15℃, 15 minutes; maintain at -15℃, 60 minutes; from -15℃ to -5℃, 10 minutes; maintain at -5℃, 120 minutes; from -5℃ to 0℃, 5 minutes; maintain at 0℃, 240 minutes.

[0054] Furthermore, the above step (3) is specifically as follows:

[0055] Weigh BFN-mEV and pure water, and dissolve and dilute them by magnetic stirring; weigh xanthan gum and pure water, and dissolve them by magnetic stirring to prepare a solution containing 0.4% xanthan gum; weigh the prescribed amount of polysorbate 80 according to the prescription, place it in a beaker, and heat it in a water bath at 28°C until it becomes transparent (if necessary), add the diluted BFN-mEV to the prescribed amount, and stir magnetically for 2 hours at a speed of about 400 rpm (adjust the room temperature to below 28°C by air conditioning during stirring) to prepare an inclusion solution; slowly add the prescribed amount of pullulan to the inclusion solution while stirring, add 0.4% xanthan gum solution to the prescribed amount, continue to weigh the prescribed amount of sucralose and mEV, add the inclusion solution, and add the remaining water to make up to volume, and continue stirring at 400 rpm for 5 minutes.

[0056] Furthermore, the loading of bugrafuran into mEVs comprises:

[0057] The buguefuran solution was mixed with the milk exosome (mEV) solution and then heated and sonicated;

[0058] In the bugrafuran solution, the solvent is anhydrous ethanol;

[0059] In the mEV solution, the solvent is 1× PBS buffer;

[0060] After the mEV is mixed with the bugrafuran solution, the final ethanol concentration does not exceed 30% v / v;

[0061] The heating conditions include: 60° C., heating treatment for 5 hours;

[0062] The ultrasonic treatment conditions include: 99% power, 20° C., and ultrasonic treatment for 18 hours.

[0063] Furthermore, the ultrafiltration concentration of the bugrafuran-mEV stock solution includes:

[0064] After dissolving the bugrafuran-mEV stock solution in PBS buffer containing PEG3350, the solution was concentrated to a volume of 1 / 20 of the stock solution using a hollow fiber column with a molecular weight cut-off of 750kDa (flow rate of 405mL / min). Experiments have shown that ultrafiltration and concentration in this step have important positive significance for increasing the loading capacity of the drug. The present invention optimizes the parameters of ultrafiltration and concentration to obtain the best effect, thereby further improving the loading effect.

[0065] Furthermore, the shearing in step (2) is carried out in an emulsifier;

[0066] Furthermore, the degassing in step (3) is carried out in an emulsifier or a degassing bottle.

[0067] Furthermore, the shearing speed is 100-3000 rpm, and the shearing time is 3-20 minutes; preferably, the emulsifier speed is 2500 rpm, and the shearing time is 10 minutes.

[0068] Furthermore, the pre-freezing temperature is -60 to -80°C; preferably, the pre-freezing temperature is -60°C.

[0069] Furthermore, the pre-freezing time is 10-100 minutes; preferably, the pre-freezing time is 15 minutes.

[0070] The present invention also provides the use of the above-mentioned mEV-encapsulated bugafuran compound nanoparticles, or the above-mentioned bugafuran freeze-dried orally disintegrating tablets in the preparation of drugs for preventing, treating or alleviating anxiety disorders.

[0071] In the present invention, the mEV-encapsulated bugafuran compound nanoparticles or the bugafuran freeze-dried orally disintegrating tablets can be used alone as a drug for preventing, treating or alleviating anxiety disorders, or in combination with other drugs.

[0072] The present invention provides a freeze-dried orally disintegrating tablet of bugrafuran, which is prepared using freeze-drying technology. The resulting product has good stability, good dissolution effect, and high sublingual absorption efficiency. The preparation method of the orally disintegrating tablet is simple and easy to implement. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 shows the electron microscopy results of mEV without additives;

[0074] Figure 2 shows the electron microscopy results of mEVs in the presence of 0.5% PEG3350 additive;

[0075] Figure 3 shows the electron microscopy results of mEVs in the presence of 5% PEG3350 additive;

[0076] FIG4 is a full view of the bugrafuran-mEV freeze-dried orally disintegrating tablet;

[0077] FIG5 is a cross-sectional view of a bugrafuran-mEV freeze-dried orally disintegrating tablet;

[0078] Figure 6 shows the blood drug concentration-time curve of bugrafuran-mEV freeze-dried orally disintegrating tablets in beagle dogs. DETAILED DESCRIPTION

[0079] In order to more clearly understand the technical content of the present invention, the following embodiments are given in detail, the purpose of which is only to better understand the content of the present invention but not to limit the scope of protection of the present invention.

[0080] The reagents used in the examples of the present invention are all commercially available.

[0081] Example 1 Preparation of a high-concentration stock solution of mEV loaded with buprenorphine

[0082] Objective: After loading buguiarfuran (BFN) into mEVs, the mEV solution was concentrated by tangential flow ultrafiltration while maintaining the stability of mEV particle number and structure.

[0083] Materials: Milk exosomes (mEVs) were prepared according to patent ZL202110097550.X.

[0084] method:

[0085] 1. Loading:

[0086] (1) Tween80 additive chemical loading method

[0087] a. 1E+13 mEV particles were diluted with Tween 80 solution to a final volume of 70% v / v with 1× PBS and preheated at 60°C for 15 min. The mEV-Tween 80 dilution was added to 150 mg of a prepared 20 mg / mL bugafuran solution (bugafuran is BFN, dissolved in anhydrous ethanol) to a final volume of 100% v / v. The sample was mixed by inversion to obtain a final ethanol concentration of 30% v / v, a final BFN concentration of 6 mg / mL, and a final Tween 80 concentration of 5% v / v. This BFN-mEV solution was obtained and designated as A.

[0088] b. At the same time, prepare a set of mEV blank controls without BFN using the same procedure.

[0089] c. After the above samples are prepared, place the samples in a 60°C water bath and heat continuously for 5 hours.

[0090] d. After heating, place the sample in a large ultrasonic cleaning machine, set the power to 99%, the temperature to 20°C, and the ultrasonic treatment time to 18 hours.

[0091] (2) BFN-mEV hybrid chemical loading method

[0092] a. Dilute the anhydrous ethanol solution with 1× PBS to a final volume of 86.5% v / v, resulting in an ethanol concentration of 20% v / v. Heat the solution at 60°C for 15 min. Add the 20% v / v ethanol dilution to 150 mg of the prepared 20 mg / mL bugafuran solution and mix thoroughly by inversion. Next, add the ethanol-BFN mixture to the mEV containing 1E+13 particles to a final volume of 100% v / v. This results in a final ethanol concentration of 30% v / v and a final BFN concentration of 2.54 mg / mL. This yields one set of BFN-mEV solution samples, designated B.

[0093] b. At the same time, prepare a set of mEV blank controls without BFN using the same procedure.

[0094] c. After the above samples are prepared, place the samples in a 60°C water bath and heat continuously for 5 hours.

[0095] d. After heating, place the sample in a large ultrasonic cleaning machine, set the power to 99%, the temperature to 20°C, and the ultrasonic treatment time to 18 hours.

[0096] (3) Hybrid chemical loading of mEVs into BFN

[0097] a. Dilute 1E+13 mEV particles with 1× PBS to a final volume of 70% v / v. Preheat at 60°C for 15 min. Add the mEV dilution to a 20 mg / mL bugafuran solution containing 150 mg of the prepared solution to a final volume of 100% v / v. Mix by inversion. The final ethanol concentration in the sample is 30% v / v, and the final BFN concentration is 6 mg / mL. This BFN-mEV solution is sample C.

[0098] b. At the same time, prepare a set of mEV blank controls without BFN using the same procedure.

[0099] c. After the above samples are prepared, place the samples in a 60°C water bath and heat continuously for 5 hours.

[0100] d. After heating, place the sample in a large ultrasonic cleaning machine, set the power to 99%, the temperature to 20°C, and the ultrasonic treatment time to 18 hours.

[0101] 2. Hollow fiber column tangential flow ultrafiltration to remove free BFN and concentrate it:

[0102] (1) Pre-experimental treatment:

[0103] Preparation of 10% m / v PEG3350: Take 60 mL of 50% PEG3350 stock solution and dilute to a final volume of 300 mL with water.

[0104] The loaded BFN-mEV stock solution sample (450 mL, particle concentration 1.6E+13 p / mL) was divided into three equal parts:

[0105] a. In a single aliquot designated 0% PEG3350-EV, take 150 mL of the BFN-mEV stock solution and add 1x PBS to a final volume of 400 mL.

[0106] b. A portion, designated 0.5% PEG3350-EV, was prepared by adding 20 mL of 10% m / v PEG3350 to 150 mL of the BFN-mEV stock solution and then calibrating the volume to a final volume of 400 mL with PBS. The final PEG3350 concentration was 0.5%.

[0107] c. In one aliquot, designated 5% PEG3350-EV, add 200 mL of 10% m / v PEG3350 to 150 mL of the BFN-mEV stock solution and bring the volume to a final volume of 400 mL with PBS. This gives a final PEG3350 concentration of 5%.

[0108] (2) Hollow fiber column tangential flow ultrafiltration concentration:

[0109] a. Cleaning Cytiva AKTA FLUX: Wash the hollow fiber column (MWCO = 750 kDa) with purified water at a flow rate of 405 mL / min until the pH of the filtrate is around 7.4.

[0110] b. Start concentration at a flow rate of 405 mL / min.

[0111] c. Concentrate to 20 mL.

[0112] d. Add 100 mL of PBS buffer containing PEG3350 each time and concentrate and filter 10 times.

[0113] e. Concentrate to 20 mL and collect the concentrate.

[0114] 3. Particle detection:

[0115] The obtained mEV-BFN product samples were tested for particle number using the Xiamen Fuliu nanoFCM nanoflow cytometer.

[0116] 4. BFN detection:

[0117] The mEV-BFN stock solution was added to methanol (final methanol concentration: 90% v / v) and centrifuged at 12,000 g for 30 minutes. The supernatant was analyzed by high-performance liquid chromatography (HPLC). The column was an Agilent ZORBAX Extend-C18; the mobile phase was 90% (v / v) methanol in water. The detection wavelength was 202 nm. BFN concentrations in the samples were calculated using an area-normalized algorithm and compared to a standard.

[0118] result:

[0119] 1. Loading:

[0120] Comparing the single-particle loading capacity of mEVs onto BFN using different loading methods, the highest and similar loading capacity was achieved in Groups B / C (i.e., samples loaded with BFN onto mEVs using a mixed chemical loading method and mEV onto BFN using a mixed chemical loading method). Approximately 17,000,000 BFN molecules could be loaded onto a single mEV particle using both loading methods. In Group A (samples loaded using a Tween 80 additive chemical loading method), the single-particle loading capacity of mEVs onto BFN was an order of magnitude lower than that achieved using the other two loading methods. The specific results are shown in Table 1.

[0121] Table 1 Comparison of mEV loading capacity on BFN under different loading methods

[0122] 2. Results of tangential flow ultrafiltration using hollow fiber columns:

[0123] The BFN-mEV stock solution had a concentration of 6.0E+12 particles / mL and a total particle count of 2.4E+15 particles before concentration. After concentration by tangential flow ultrafiltration on a hollow fiber column in the presence of 0.5% and 5% PEG3350 additives, the total particle count was more than two-fold higher than that obtained with 0% PEG335. With 5% PEG3350, the particle count reached a maximum of 1.58E+15 particles, with a concentration of 7.88E+13 particles / mL. The resulting total particle count accounted for 65.7% of the total pre-concentration count, representing a 13-fold increase in concentration. Electron microscopy revealed that mEV dispersion was improved in the presence of 5% PEG3350. The results are shown in Table 2.

[0124] Table 2 Concentration effect of BFN-mEV at different concentrations of PEG3350

[0125] Figure 1 shows the electron microscopy results of mEV without additives; Figure 2 shows the electron microscopy results of mEV in the presence of 0.5% PEG3350 additive; Figure 3 shows the electron microscopy results of mEV in the presence of 5% PEG3350 additive.

[0126] Example 2 Preparation of Bugrafuran-mEV Freeze-dried Orally Disintegrating Tablets

[0127] 1. Preparation of BFN-mEV Lyophilized Orally Disintegrating Tablets

[0128] Using bugrafuran (BFN) loaded into mEV as raw material, BFN-mEV freeze-dried orally disintegrating tablets were prepared according to the formulation shown in Table 3 below.

[0129] Table 3 BFN-mEV freeze-dried orally disintegrating tablets formulation (calculated based on pre-freeze-drying mass)

[0130] Specific steps:

[0131] (1) Weigh BFN-mEV and pure water, and dissolve and dilute them by magnetic stirring; weigh xanthan gum and pure water, and dissolve them by magnetic stirring to prepare a solution containing 0.4% xanthan gum; weigh the prescribed amount of polysorbate 80 according to the prescription, place it in a beaker, heat it in a water bath at 28°C until it becomes transparent (if necessary), add the diluted BFN-mEV to the prescribed amount, and stir magnetically for 2 hours at a speed of about 400 rpm (adjust the room temperature to below 28°C by air conditioning during stirring) to prepare an inclusion solution; slowly add the prescribed amount of pullulan to the inclusion solution while stirring, add 0.4% xanthan gum solution to the prescribed amount, continue to weigh the prescribed amount of sucralose and mEV, add them to the inclusion solution, and add the remaining water to make up the volume, and continue stirring at 400 rpm for 5 minutes.

[0132] (2) Shearing: Use an emulsifier for shearing, speed: 2500 rpm, time: 10 minutes.

[0133] (3) Degassing: Transfer the drug solution to a 125 ml degassing bottle and vacuum degas until there are no bubbles.

[0134] (4) Filling: Use a pipette to fill into a 0.4 ml aluminum container, 0.4 g / tablet.

[0135] (5) Prefreezing: Place the filled aluminum nests in a -60°C low-temperature refrigerator for prefreezing for 20 minutes. After prefreezing, store them in the refrigerator until they are placed in the freeze dryer.

[0136] (6) Freeze-drying process: transfer the pre-frozen sample to 1.7m 2 Freeze drying was performed in a freeze dryer, and the freeze drying curve is shown in Table 4 below:

[0137] Inlet temperature: -30℃; Cold trap temperature: ≤-40℃.

[0138] Table 4 Freeze-drying procedure

[0139] The full view and cross-sectional view of the prepared bugrafuran-mEV freeze-dried orally disintegrating tablets are shown in Figures 4 and 5, respectively.

[0140] The prepared bugrafuran-mEV freeze-dried orally disintegrating tablets were evaluated, and the results are shown in Table 5:

[0141] Table 5 Evaluation results

[0142] The full view and cross-sectional view of the film are shown in Figures 4 and 5.

[0143] 2. 6-month stability evaluation of active ingredients

[0144] Table 6 6-month stability evaluation of active ingredients

[0145] Example 3 Animal Pharmacokinetic Study of Bugafuran-mEV Lyophilized Orally Disintegrating Tablets

[0146] Objective: To evaluate the plasma concentration and exposure of bugrafuran-mEV freeze-dried orally disintegrating tablets in beagle dogs after oral administration.

[0147] Test method:

[0148] a. The test groups are shown in Table 7:

[0149] Table 7 Test groups

[0150] b. Blood collection time: Blood was collected at 0.5h, 1h, 2h, 4h, 8h, 12h, and 24h after administration.

[0151] c. Detection: The BFN concentration of blood samples was detected by LC-MS method.

[0152] result:

[0153] The plasma concentration-time curves of bugrafuran-mEV freeze-dried orally disintegrating tablets in beagle dogs are shown in Figure 6. Specific data are shown in Table 8.

[0154] Table 8 Animal pharmacokinetic test results

[0155] BLQ: below the detection limit

[0156] The animal blood drug concentration results showed that compared with the negative control without mEV, the oral bioavailability of BFN orally disintegrating tablets loaded with mEVs was significantly improved, and the peak time was delayed. A lower dose could reach or even exceed the blood drug concentration that could be achieved by the original traditional oral preparation, and it had better pharmacokinetic properties.

[0157] Example 4 Taste Evaluation of Bugafuran-mEV Freeze-Dried Orally Disintegrating Tablets

[0158] Objective: To evaluate the taste and mouthfeel of bugrafuran-mEV freeze-dried orally disintegrating tablets and bugrafuran freeze-dried orally disintegrating tablets.

[0159] Test method:

[0160] Five volunteers were recruited and administered the freeze-dried orally disintegrating tablets on day one and day five. Feedback was recorded regarding mouthfeel, flavor, and disintegration and absorption rates. Each volunteer was unaware of the composition of the freeze-dried orally disintegrating tablets they were given. The order of administration was randomized: either BFN-mEV or BFN freeze-dried orally disintegrating tablets were administered on day one, followed by the other on day five.

[0161] result:

[0162] Three of the five volunteers thought that the dissolution rate of BFN-mEV freeze-dried orally disintegrating tablets was slightly slower than that of BFN freeze-dried orally disintegrating tablets, and two thought there was no difference; four thought that BFN-mEV freeze-dried orally disintegrating tablets had no bitter taste and a slight milky aroma, and one thought that there was no obvious taste; all five volunteers thought that BFN freeze-dried orally disintegrating tablets tasted bitter.

[0163] The above results show that BFN-mEV freeze-dried orally disintegrating tablets are superior to BFN freeze-dried orally disintegrating tablets in terms of taste.

[0164] The above embodiments are only intended to help understand the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by those skilled in the art, and such improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A bugrafuran drug freeze-dried orally disintegrating tablet, characterized in that: It contains the following raw materials in percentage by weight: 10%-90% bugafuran-mEV stock solution, 1%-5% binder, 0.1%-2% suspending agent, 0.1%-1% surfactant, 0.1%-1% sweetener, 0.1%-1% flavoring agent, and the balance is water; The surfactant is selected from polysorbate 80.

2. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The adhesive is selected from one of pullulan and gelatin; preferably, the adhesive is pullulan.

3. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The suspending agent is selected from one of gum arabic, gum tragacanth, peach gum and xanthan gum; preferably, the suspending agent is xanthan gum.

4. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The sweetener is selected from one of sucralose, sucrose, steviol glycoside, and saccharin sodium; preferably, the sweetener is sucralose.

5. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The flavoring agent is selected from one of sweet orange flavor, strawberry flavor, mEV, cherry flavor, and tangerine flavor; preferably, the flavoring agent is mEV.

6. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The raw materials for preparing the bugrafuran-mEV stock solution include: adding 1×10 10 ~1×10 13 mEV particles.

7. The bugrafuran drug freeze-dried orally disintegrating tablet according to claim 1, characterized in that: The bugrafuran is bugrafuran or its in vivo metabolite.

8. The bugrafuran drug freeze-dried orally disintegrating tablet according to any one of claims 1 to 7, characterized in that: The invention comprises the following raw materials in percentage by mass: 10%-90% bugafuran-mEV stock solution, 1%-5% pullulan, 0.1%-2% xanthan gum, 0.1%-1% polysorbate 80, 0.1%-1% sucralose, 0.1%-1% mEV, and the balance is water.

9. The method for preparing the freeze-dried orally disintegrating tablet of bugrafuran drug according to any one of claims 1 to 8, characterized in that: The steps include: (1) Loading bucuran onto mEVs to obtain bucuran-mEV stock solution; (2) The bucuronium-mEV stock solution was concentrated by ultrafiltration to obtain a concentrated solution; (3) mixing the concentrated solution with a surfactant in a solution to obtain an inclusion solution; mixing the inclusion solution with a binder, a suspending agent, a sweetener, and a flavoring agent to obtain a drug solution; (4) shearing the liquid medicine in step (3); (5) degassing the liquid medicine after shearing in step (4); (6) filling the degassed liquid medicine in step (5) into the cavity; (7) Pre-freeze the filled liquid medicine; (8) Freeze-dry the pre-frozen drug, setting the initial temperature to -30°C. The specific freeze-drying process is as follows: Cold soaking temperature: ≤-60℃; Vacuum control: ≤200ubar Plate temperature: from -30℃ to -15℃, 15 minutes; maintained at -15℃, 60 minutes; from -15℃ to -5℃, 10 minutes; maintained at -5℃, 120 minutes; from -5℃ to 0℃, 5 minutes; maintained at 0℃, 240 minutes.

10. The preparation method according to claim 9, characterized in that: The loading of bugrafuran into mEVs comprises: The buguafuran solution and the milk exosome solution were mixed and then heated and sonicated; In the bugafuran solution, the solvent is anhydrous ethanol; In the milk exosome solution, the solvent is 1×PBS buffer; After the milk exosomes are mixed with the bugrafuran solution, the final ethanol concentration does not exceed 30% v / v; The heating conditions include: 60° C., heating treatment for 5 hours; The ultrasonic treatment conditions include: 99% power, 20° C., and ultrasonic treatment for 18 h.

11. The preparation method according to claim 9, characterized in that: The ultrafiltration concentration of the bugrafuran-mEV stock solution comprises: The bucuronium-mEV stock solution was dissolved in PBS buffer containing PEG3350 and concentrated on a hollow fiber column with a molecular weight cutoff of 750 kDa at a flow rate of 405 mL / min to a volume of 1 / 20 of the stock solution.

12. The preparation method according to claim 9, characterized in that: The shearing speed is 100-3000 rpm, and the shearing time is 3-20 minutes; preferably, the emulsifier speed is 2500 rpm, and the shearing time is 10 minutes.

13. The preparation method according to claim 9, characterized in that: The pre-freezing temperature is -60 to -80°C; preferably, the pre-freezing temperature is -60°C.

14. The preparation method according to claim 9, characterized in that: The pre-freezing time is 10-100 minutes; preferably, the pre-freezing time is 15 minutes.

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