阅读说明：本技术 蒙艾中黄酮类物质的微囊包合物的制备方法 (Preparation method of microcapsule inclusion compound of flavonoids in mugwort ) 是由 洪海龙 胡毛乾 石俊庭 竺宁 郝金莹 梁秀雪 于 2021-07-28 设计创作，主要内容包括：本发明涉及一种蒙艾中黄酮类物质的微囊包合物的制备方法,其包括：S1、将从蒙艾中提取纯化的黄酮类药物原料和载体混合,加入有机溶剂,经搅拌及超声处理等方式,得到透明澄清的溶液；S2、将溶液转移至超临界反应釜中,通入CO-(2)使超临界反应釜内压力保持在12MPa以上,加热使超临界反应釜内的温度保持在40-55℃,压力和温度达设定值后,对该超临界反应釜内的溶液进行搅拌；S3、搅拌结束后,缓速释放掉超临界反应釜内的溶剂,释放完毕后,采用干净气流吹干留在所述超临界反应釜内的固体物,吹干后,制得黄酮类药物的微囊包合物。该方法制备过程简单、温度低、可保持药物活性、环境友好,产物有机残留少,溶出速度等优点。(The invention relates to a preparation method of a microcapsule inclusion compound of flavonoids in mugwort, which comprises the following steps: s1, mixing the flavonoid drug raw material extracted and purified from the mugwort with a carrier, adding an organic solvent, stirring, carrying out ultrasonic treatment and the like to obtain a transparent and clear solution; s2, transferring the solution into a supercritical reaction kettle, and introducing CO 2 Keeping the pressure in the supercritical reaction kettle above 12MPa, heating to keep the temperature in the supercritical reaction kettle at 40-55 ℃, and stirring the solution in the supercritical reaction kettle after the pressure and the temperature reach set values; s3, after stirring, slowly releasing the solvent in the supercritical reaction kettle, and after releasing, blowing and drying the solvent by clean air flow to remain in the supercritical reaction kettleDrying the solid in the critical reaction kettle by blowing, and obtaining the microcapsule inclusion compound of the flavonoid drug. The method has the advantages of simple preparation process, low temperature, capability of maintaining the activity of the medicine, environmental friendliness, less organic residue of the product, high dissolution speed and the like.)

1. A preparation method of a microcapsule inclusion compound of flavonoids in mugwort is characterized by comprising the following steps:

s1, mixing the flavonoid drug material extracted and purified from the mugwort with a certain amount of carrier, adding an organic solvent, and performing one or more of stirring, oscillation and ultrasonic treatment to obtain a transparent and clear solution;

s2, transferring the solution into a supercritical reaction kettle, and introducing CO₂Keeping the pressure in the supercritical reaction kettle above 12MPa, heating to keep the temperature in the supercritical reaction kettle at 40-55 ℃, and stirring the solution in the supercritical reaction kettle after the pressure and the temperature reach set values;

and S3, after stirring, slowly releasing the solvent in the supercritical reaction kettle, after the solvent is released, blowing the solid remained in the supercritical reaction kettle by using clean air flow, and blowing to dry to obtain the microcapsule inclusion compound of the flavonoid drug.

2. The method of claim 1, wherein in S1, the flavonoid drug substance extracted and purified from Artemisia montana is quercetin, naringenin, luteolin, or apigenin.

3. The method according to claim 1, wherein the carrier is one or more selected from HPMC, HP- β -CD, PVP K-30 and SBE- β -CD in S1.

4. The preparation method according to claim 2, wherein in S1, the mass ratio of the flavonoid drug raw material to the carrier is 1: 1-3.

5. The preparation method according to claim 1, wherein in S1, when the flavonoid drug substance is quercetin, naringenin or luteolin, the organic solvent is anhydrous methanol or acetone;

when the flavonoid medicine raw material is apigenin, the organic solvent is absolute ethyl alcohol/absolute methyl alcohol and dichloromethane in a volume ratio of 1: 1.

6. The preparation method according to claim 1, wherein 1mL of the organic solvent dissolves 5-30mg of the flavonoid pharmaceutical raw material and the carrier in total weight.

7. The method according to claim 1, wherein in S2, CO is introduced into the supercritical reactor₂The pressure value is 12-18 MPa; the stirring time is 30-60min, and the rotating speed is 400-900 rpm.

8. The method according to claim 1, wherein a filter membrane capable of retaining the microencapsulated clathrate to be prepared is provided at a solvent discharge valve port of the supercritical reaction tank in S3.

9. The method of claim 1, wherein the clean gas stream is CO at S3₂Supercritical CO provided by gas source₂。

Technical Field

The invention relates to the technical field of drug sustained release, in particular to a preparation method of a microcapsule inclusion compound of flavonoids in mugwort based on a supercritical anti-solvent method.

Background

The wild mugwort in inner Mongolia area is called as "Mongolia". Mugwort contains various bioactive components including various flavonoids. Typical flavonol substances obtained by separating and purifying the Artemisia princeps comprise quercetin, luteolin, naringenin, apigenin and the like, and the substances have high medicinal values. However, the flavonoid has a C6-C3-C6 planar mother ring structure of 2-phenylchromone, so that the flavonoid has poor water solubility, has low bioavailability after entering a biological organism, and has certain toxicity to organism cells when the flavonoid is excessively accumulated in the biological organism. Therefore, the improvement of the water solubility of the flavonoids in the mugwort has important significance for improving the bioavailability, effectively utilizing the medicinal value and preventing toxicity. The microencapsulation technology is an important means for improving the water solubility of the medicine and improving the bioavailability.

In recent years, scholars at home and abroad also put forward some technical schemes for microencapsulation of flavonoids. For example, romboiss et al (CN105086002B) disclose spirochete dextrin quercetin inclusion complexes, prepared by a solvent evaporation method to improve the water solubility of quercetin; shangjingchuan et al (CN105031663B) disclose that SBE-beta-CD (sulfobutyl ether beta-cyclodextrin) is used as a carrier, and a physical grinding method is adopted to prepare a microcapsule inclusion compound of luteolin so as to improve water solubility; the Lushengmen et al (CN111493250A) disclose that the solubility and bioavailability of naringin can be improved by using octenyl succinic waxy corn starch ester (OSAS) as a wrapping material and adopting a solvent evaporation method to carry out inclusion on the naringin. Wang Zhongni et al (CN109010271A) disclose that apigenin is prepared into a microemulsion by mixing polyoxyethylene sorbitan monooleate, 1, 2-propylene glycol, geraniol and water, and the solubility of the apigenin is improved. Quercetin beta-cyclodextrin inclusion compound-chitosan microspheres prepared by solvent evaporation method, with drug loading of 12.3% and average particle size of 3.327 μm, were proposed by Hu Ying et al (10.6039/j.issn.1001-0408.2012.33.13). Guo Tonglin et al (10.3969/j.issn.1008-0805.2020.11.021) suggested that the physicochemical properties of the quercetin solid lipid nanoparticles prepared by the same were: the average particle size is 191.2nm, the drug loading is 4.33 percent, and the cumulative dissolution is 87.6 percent in 72 hours. Miao Ye et al (1006) -3765(2020) -07-0009-04) propose to prepare quercetin sustained release tablets by tabletting method with HPMC as a matrix and CMC-Na, talcum powder and starch as auxiliary materials. The slow release experiment is carried out according to the national standard, and the cumulative dissolution reaches about 80 percent in 12 hours. Dianling hui et al (1004-. The traditional method has the defects that the temperature is too high, so that the flavonoid compound is easy to inactivate, the residual quantity of the organic solvent is large, the accumulative dissolution is slow and the like.

Disclosure of Invention

Technical problem to be solved

In view of the defects and shortcomings of the prior art, the invention provides a preparation method of a micro-capsule inclusion compound of flavonoids in mugwort, which mainly adopts a supercritical anti-solvent method (SAS) to prepare flavonoid compounds in the mugwort such as quercetin, and the method has the advantages of simple preparation process, low temperature, no influence on the activity of the medicine, environmental friendliness, less organic residue and the like, and can improve the dissolution rate of the medicine and the bioavailability of the medicine.

(II) technical scheme

In order to achieve the purpose, the invention adopts the main technical scheme that:

a method for preparing microcapsule inclusion compound of flavonoid in Artemisia Mongolica comprises:

s1, mixing the flavonoid drug material extracted and purified from the mugwort with a certain amount of carrier, adding an organic solvent, and performing one or more of stirring, oscillation and ultrasonic treatment to obtain a transparent and clear solution;

s2, transferring the solution into a supercritical reaction kettle, and introducing CO₂Keeping the pressure in the supercritical reaction kettle above 12MPa, heating to keep the temperature in the supercritical reaction kettle at 40-55 ℃, and stirring the solution in the supercritical reaction kettle after the pressure and the temperature reach set values;

and S3, after stirring, slowly releasing the solvent in the supercritical reaction kettle, after the solvent is released, blowing the solid remained in the supercritical reaction kettle by using clean air flow, and blowing to dry to obtain the microcapsule inclusion compound of the flavonoid drug.

According to a preferred embodiment of the present invention, in S1, the flavonoid drug material extracted and purified from artemisia mongolica is quercetin, naringenin, luteolin or apigenin.

According to a preferred embodiment of the present invention, in S1, the carrier is one or a combination of HPMC, HP-beta-CD, PVP K-30 and SBE-beta-CD. For example, in the preparation of the quercetin microcapsule compound, the carrier may be hydroxypropyl methylcellulose (HPMC), polyvinylpyrrolidone (PVP K-30) in a ratio of 1.5: 1.

According to the preferred embodiment of the invention, in S1, the mass ratio of the flavonoid drug raw material to the carrier is 1: 1-3; preferably 1:2 to 2.5.

According to the preferred embodiment of the present invention, in S1, when the flavonoid drug material is quercetin, naringenin or luteolin, the organic solvent is anhydrous methanol or acetone; when the flavonoid medicine raw material is apigenin, the organic solvent is absolute ethyl alcohol/absolute methyl alcohol and dichloromethane in a volume ratio of 1: 1.

According to a preferred embodiment of the present invention, in S1, the organic solvent is used in an amount of: 1mL of organic solvent is used for dissolving flavonoid drug raw materials and carriers with the total weight of 5-30mg, so that transparent and clear solution can be obtained, and the concentration of the solution is not too low, thereby causing low preparation efficiency.

Preferably, after the organic solvent is added in S1, a transparent and clear solution is obtained through stirring and ultrasonic treatment; the ultrasonic power is 125 kW/h.

According to the preferred embodiment of the present invention, in S2, CO is introduced into the supercritical reactor₂The pressure value is 12-18MPa, preferably 14 MPa; heating to keep the temperature in the supercritical reaction kettle at 45 ℃; the stirring time is 30-60min, and the rotating speed is 400-900 rpm. Preferably, the stirring time is 30min and the rotation speed is 500 rpm.

Preferably, CO is arranged outside the supercritical reaction kettle₂Source of gas, CO₂The gas source is connected with the supercritical reaction kettle through a pressurizer (such as a diaphragm pressurizer), and the pressure in the supercritical reaction kettle reaches a set value by adjusting the output pressure of the pressurizer; and a heating device is arranged outside or at the bottom of the supercritical reaction kettle, and the heating device can accurately control the temperature in the supercritical reaction kettle. In order to facilitate observation, an observation window can be arranged on the supercritical reaction kettle, or the supercritical reaction kettle is made of a pressure-resistant transparent material. Quadrant change in the reaction kettle can be observed through a window and the like, and the solvent is emptied.

According to the preferred embodiment of the present invention, in S3, the solvent in the supercritical reactor is slowly released, so as to avoid blowing out the microcapsule encapsulated substance due to too fast releasing speed of the solvent. Or a filter membrane is arranged at the solvent application valve of the supercritical reaction kettle and can intercept the prepared microcapsule inclusion compound.

According to the preferred embodiment of the present invention, in S3, the clean gas stream is CO₂Supercritical CO provided by gas source₂。

(III) advantageous effects

Compared with the prior art, the invention has the following technical effects:

(1) when the microcapsule inclusion compound of flavonoids in mugwort is prepared, the temperature is not more than 55 ℃ at most, and the flavonoid drugs are not inactivated due to high-temperature heating.

(2) The dosage of the organic solvent used in the invention is very low, and each 1mL of the organic solvent dissolves the flavonoid drug raw materials and the carrier with the total weight of 5-30mg, so the dosage of the organic solvent is small. After the reaction in the supercritical reaction kettle is finished, when the organic solvent is released, supercritical CO is adopted₂Most of the organic solvent in the microcapsule inclusion compound is taken away. Finally passing through supercritical CO₂And further drying the substances left in the reaction kettle to remove the solvent. Therefore, the residual solvent quantity in the flavonoid microcapsule inclusion compound prepared by the invention is extremely low and can be controlled to be below 10 ppm.

(3) The product prepared by the method of the invention is subjected to infrared spectrum testing. The test results demonstrate that the drug has indeed been loaded on the carrier material. Through the analysis of the combination of a Scanning Electron Microscope (SEM) and particle size analysis, the microcapsule inclusion compound prepared by the method has the average particle size of 23.67-71.20 microns, the drug loading rate of 7.06-70.13 percent and the drug loading rate is obviously higher than that of nano particles prepared by a film dispersion method and a solid lipid method in the prior art.

(4) Further testing the drug dissolution release rate of the microcapsule inclusion compound, the maximum cumulative dissolution of quercetin at 140min can reach 53.95%, the maximum cumulative dissolution of naringenin at 140min can reach 81.4%, the maximum cumulative dissolution of luteolin at 140min can reach 51.93%, and the maximum cumulative dissolution of apigenin at 140min can reach 29.68%. Therefore, the medicine dissolution speed of the microcapsule inclusion compound prepared by the invention is very high, so that the solubility of the total flavonoid compounds in the mugwort is greatly improved, and the bioavailability of the total flavonoid compounds is improved.

Drawings

FIG. 1 is a schematic diagram of an apparatus for preparing microcapsule inclusion compound of flavonoid in Artemisia Mongolica.

FIG. 2 is a comparison graph of infrared spectra of quercetin-HPMC-PVP ternary microcapsule inclusion compound, HPMC, PVP and quercetin.

Fig. 3 is an SEM image of the quercetin-HPMC-PVP ternary microcapsule inclusion compound of example 1.

Fig. 4 is an SEM image of the quercetin-HPMC-PVP ternary microcapsule inclusion compound.

FIG. 5 is an SEM photograph of the luteolin-HPMC-PVP ternary microencapsulation complex of example 4.

Fig. 6 is an SEM image of naringenin-HPMC-PVP ternary microencapsulation complex of example 5.

Fig. 7 is an SEM image of the apigenin-HPMC-PVP ternary microencapsulation complex of example 6.

Fig. 8 is an SEM image of the quercetin-HP- β -CD binary microencapsulation inclusion compound of example 10.

Fig. 9 is a comparison of the cumulative dissolution release profiles of the physical mixtures of quercetin-HPMC binary microcapsule inclusion compound, pure quercetin, and quercetin-HPMC prepared in example 12.

FIG. 10 is a graph of particle size analysis of the Quercetin-HP- β -CD binary clathrate prepared in example 10.

Fig. 11 is a cumulative dissolution profile of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 1.

Fig. 12 is a cumulative dissolution profile of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 2.

Fig. 13 is a cumulative dissolution profile of the luteolin-HPMC-PVP ternary microencapsulation complex prepared in example 4.

Fig. 14 is a cumulative dissolution profile of the apigenin-HPMC-PVP ternary microencapsulation inclusion compound prepared in example 6.

Fig. 15 is a cumulative dissolution profile of the naringenin-HPMC-PVP ternary microencapsulation complex prepared in example 7.

Fig. 16 is a cumulative dissolution profile of the quercetin HP- β -CD binary micro-capsule clathrate prepared in example 10.

Fig. 17 is a graph of particle size analysis of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 2.

Fig. 18 is a particle size analysis diagram of the luteolin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 4.

Fig. 19 is a particle size analysis diagram of the naringenin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 5.

Fig. 20 is a particle size analysis diagram of the apigenin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 6.

Detailed Description

For the purpose of better explaining the present invention and to facilitate understanding, the present invention will be described in detail by way of specific embodiments with reference to the accompanying drawings.

As shown in fig. 1, the visible reaction kettle 10 is provided with three valves 1,2, 3. The valve 1 is connected with a carbon dioxide gas source. Carbon dioxide is introduced into the visible reaction kettle 10 from a carbon dioxide gas source through a diaphragm compressor, so that the pressure of the visible reaction kettle 10 reaches more than 12MPa (preferably between 12 and 18 MPa). A magnetic stirring device and a heating device are arranged below the visual reaction kettle 10 and are respectively used for stirring and heating the materials in the visual reaction kettle 10 and maintaining a proper reaction temperature. The valve 2 is mainly used for blowing clean air flow such as supercritical carbon dioxide to blow dry materials in the visible reaction kettle 10 after the solvent is applied when the microcapsule compound is prepared. The valve 3 is used for dispensing solvent.

The following examples will use the apparatus shown in figure 1 to prepare the microencapsulated inclusion compound.

Example 1

Weighing 501.5mg of quercetin, 750.7mg of hydroxypropyl methylcellulose (HPMC) and 500.5mg of polyvinylpyrrolidone (PVP K-30), adding all the materials into a beaker, adding 80mL of anhydrous methanol, stirring uniformly, dispersing by using ultrasound to obtain a uniform and transparent solution, adding the solution into a supercritical visible reaction kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, and setting 500rpm to start timing for 30 min. After the stirring is finished, the stirring is stopped and the solvent is released. And observing quadrant change in the window until the solvent is completely released. Introducing supercritical carbon dioxide (SC-CO)₂) And (4) drying for 1h, and obtaining 842.9mg of the quercetin microcapsule inclusion compound from the reaction kettle, namely the quercetin-HPMC-PVP ternary microcapsule inclusion compound.

The prepared microcapsule inclusion compound was detected by infrared spectroscopy as shown in fig. 2, and the spectrum shown in fig. 2 was obtained. Meanwhile, pure HPMC, pure PVP and pure quercetin were subjected to infrared detection, and the infrared spectrograms of the microcapsule inclusion compound, HPMC, PVP and quercetin prepared in example 1 were compared. The infrared ray of the HPMC, PVP, quercetin and-HPMC-PVP ternary microcapsule inclusion compound is sequentially corresponding to the infrared ray of the HPMC, PVP, quercetin and-HPMC-PVP ternary microcapsule inclusion compound from bottom to top in the figureSpectrum of light. By comparison, in the infrared spectrogram of the carrier (hydroxypropyl methylcellulose HPMC, polyvinylpyrrolidone (PVP K-30)) and the bulk drug quercetin, the phenolic hydroxyl group of the bulk drug quercetin is red-shifted and the peak shape thereof is widened, which indicates that the phenolic hydroxyl group is associated, and at the same time, the peak length is 1650cm^-1The ketone carbonyl characteristic peak disappears, which proves that the drug is bonded with the carrier, and the carrier wraps the drug quercetin.

As shown in fig. 3, which is an SEM image of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in example 1, fig. 4 is an SEM image of the quercetin-HPMC-PVP ternary microcapsule inclusion compound at a higher magnification. The figure shows that the porous structure formed by the disordered winding of the micro fibers has high specific surface area. The drug loading rate of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in this example was 25.5%, and the cumulative dissolution curve is shown in FIG. 11. The 130min cumulative dissolution rate is 32.0%.

Example 2

Weighing 103.5mg of quercetin, 150.9mg of hydroxypropyl methylcellulose (HPMC) and 100.1mg of polyvinylpyrrolidone (PVP K-30), adding into a beaker, adding 40mL of anhydrous methanol, stirring, and dispersing with ultrasound to obtain a uniform and transparent solution. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring after timing is finished. Releasing solvent, observing quadrant change in the window, introducing supercritical carbon dioxide (SC-CO) after the solvent is completely released₂) Blow-drying, and after timing for 1h, obtaining 210.7mg of the quercetin microcapsule inclusion compound, namely the quercetin-HPMC-PVP ternary microcapsule inclusion compound, from the kettle. Scanning by an electron microscope is carried out, and the scanning result is shown in figures 3-4. The drug loading rate of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in the embodiment is 25.98%, and the cumulative dissolution curve is shown in figure 12; the 130min cumulative dissolution rate is 34.75%; the average particle size of the inclusion compound was 71.27 μm (see FIG. 17 for a particle size analysis chart).

Example 3

Weighing quercetin 100.6mg, hydroxypropyl methylcellulose (HPMC)150.4mg, and polyvinylpyrrolidone (PVP K-30)100.5mg, adding into beaker, and adding40mL of acetone was stirred well and dispersed into a uniform clear solution using sonication. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 17MPa, starting stirring after the temperature and the pressure reach set values, and setting 500rpm to start timing for 30 min. After the timing was completed, the stirring was stopped and the solvent was released. Observing quadrant change in the window, releasing solvent completely, and introducing supercritical carbon dioxide (SC-CO)₂) Blow-drying, and after timing for 1h, obtaining 210.3mg of quercetin microcapsule inclusion compound from the kettle. The drug loading rate of the quercetin-HPMC-PVP ternary microcapsule inclusion compound prepared in the embodiment is 27.46%.

Example 4

100.6mg of luteolin, 150.7mg of hydroxypropyl methylcellulose (HPMC) and 100.1mg of polyvinylpyrrolidone (PVP K-30) were weighed into a beaker, added with 40mL of anhydrous methanol, stirred and dispersed with ultrasound to form a uniform and transparent solution. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm, starting timing for 30min, stopping stirring after timing is finished, and starting to release the solvent. Observing quadrant change in the window, releasing solvent completely, and introducing supercritical carbon dioxide (SC-CO)₂) Blow-drying, and obtaining 136.6mg of microcapsule inclusion compound from the kettle after timing 1 h.

As shown in fig. 5, is an SEM image of the luteolin-HPMC-PVP ternary microencapsulation complex prepared in this example. As can be seen from the figure, the luteolin and the HPMC and other carriers form a coralline porous three-dimensional structure, and form a microcapsule inclusion compound which is good and has a large specific surface area. The drug loading of the luteolin in the luteolin-HPMC-PVP ternary microcapsule inclusion compound is detected to be 17.93%.

The luteolin-HPMC-PVP ternary microcapsule inclusion compound is tested for dissolution, and the test result shows that (shown as a cumulative dissolution curve in figure 13): at any moment, the dissolution rate of the luteolin-HPMC-PVP ternary microcapsule inclusion compound is far higher than that of a physical mixture of the luteolin and the luteolin/HPMC/PVP, and the cumulative dissolution can reach 51.93% in 140 min. As shown in figure 18, the luteolin-HPMC-PVP ternary microcapsule clathrate has a particle size analysis chart, and the average particle size DAV is 71.19 μm.

Example 5

Naringenin 100.7mg, Hydroxypropylmethylcellulose (HPMC)150.2mg, polyvinylpyrrolidone (PVP K-30)100.5mg were weighed and added to a beaker with 40mL of anhydrous methanol. Stirred well and dispersed into a uniform transparent solution using ultrasound. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring and starting to release the solvent after timing is finished. Observing quadrant change in the window, releasing solvent completely, and introducing supercritical carbon dioxide (SC-CO)₂) After the blow-drying time is over for 1h, 105.4mg of microcapsule inclusion compound is obtained from the kettle.

As shown in fig. 6, is SEM image of naringenin-HPMC-PVP ternary microencapsulation inclusion compound. It can be seen from the figure that the naringenin and the carrier form a particle-piled porous three-dimensional structure to form a good microcapsule inclusion compound with a large specific surface area. The medicine-loading rate of naringenin in the naringenin-HPMC-PVP ternary microcapsule inclusion compound prepared in this example is 21.53%. FIG. 19 shows the particle size analysis chart of naringenin-HPMC-PVP ternary microcapsule clathrate, with the average particle size DAV of 71.20 μm.

Example 6

50.8mg of apigenin, 75.5mg of hydroxypropyl methylcellulose (HPMC) and 50.3mg of polyvinylpyrrolidone (PVP K-30) are weighed, added into a beaker, added with 20mL of anhydrous methanol and 20mL of dichloromethane (175mg:40mL), stirred uniformly and dispersed by using ultrasound to form a uniform and transparent solution. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 deg.C and the pressure at 14MPa, starting stirring and setting 500rpm after the temperature and pressure reach the set value, starting timing for 30min, stopping stirring after timing, starting to release the solvent and observing quadrant change in a window, completely releasing the solvent, and introducing supercritical carbon dioxide (SC-CO)₂) Blow-drying, and obtaining 61.3mg of microcapsule inclusion compound from the kettle after timing 1 h.

As shown in fig. 7, is an SEM image of the apigenin-HPMC-PVP ternary microcapsule inclusion compound. The figure shows that the apigenin and the carrier form a good microcapsule inclusion compound with a large specific surface area. The drug loading rate of the apigenin in the apigenin-HPMC-PVP ternary microcapsule inclusion compound prepared in the embodiment is 11.79%.

The dissolution rate of the ternary microcapsule inclusion compound is tested, and the test result shows that: at any time, the dissolution rate of the apigenin-HPMC-PVP ternary microcapsule inclusion compound is higher than that of a physical mixture of apigenin and apigenin/HPMC/PVP, and the cumulative dissolution rate at 130min is 38.73% (as shown in figure 14). The particle size analysis of the apigenin microcapsule inclusion compound is shown in fig. 20, and the average particle size is Dav 68.76 μm.

Example 7

Weighing 50.3mg of naringenin, 75.3mg of hydroxypropyl methylcellulose (HPMC) and 50.3mg of polyvinylpyrrolidone (PVP K-30), adding all the naringenin, 75.3mg of hydroxypropyl methylcellulose (HPMC) and 50.3mg of polyvinylpyrrolidone (PVP K-30) into a beaker, adding 40mL of anhydrous methanol into the beaker, uniformly stirring, dispersing by using ultrasound to obtain a uniform and transparent solution, adding the solution into a supercritical visible kettle, closing the kettle, setting the temperature to be 45 ℃ and the pressure to be 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm to start timing for 30min, stopping stirring after the timing is finished, and starting to release the solvent. Observing quadrant change in the window, releasing solvent completely, and introducing supercritical carbon dioxide (SC-CO)₂) Blow-drying, and obtaining 56.1mg of microcapsule inclusion compound from the kettle after timing 1 h. The medicine-loading rate of naringenin in the naringenin-HPMC-PVP ternary microcapsule inclusion compound prepared in this example is 31.89%, the cumulative dissolution rate is shown in FIG. 15, and the maximum value of the cumulative dissolution rate reaches 82.8302%.

Example 8

Weighing 100.1mg of quercetin and 150.0mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD), and 100.4mg of polyvinylpyrrolidone (PVP K-30), adding all the materials into a beaker, adding 40mL of anhydrous methanol, uniformly stirring, dispersing by using ultrasound to obtain a uniform and transparent solution, adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature to be 45 ℃ and the pressure to be 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm to start timing for 30min, stopping stirring after the timing is finished, and starting to release the solvent. Observing quadrant change in the window, releasing solvent completely, and starting to use supercritical carbon dioxide (SC-CO)₂) Blow-dryAfter the time was 1 hour, 205.9mg of the microencapsulated product was obtained from the autoclave. The drug loading rate of the quercetin HP-beta-CD-HPMC-PVP ternary microcapsule inclusion compound prepared in the embodiment is 20.31%.

Example 9

Weighing 100.2mg of naringenin, 150.7mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD) and 100.3mg of polyvinylpyrrolidone (PVP K-30), adding all the materials into a beaker, adding 40mL of anhydrous methanol, uniformly stirring, dispersing by using ultrasonic to obtain a uniform and transparent solution, adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature to be 45 ℃ and the pressure to be 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm to start timing for 30min, stopping stirring after the timing is finished, and starting to release the solvent. Observing quadrant change in the window, releasing solvent completely, and starting to use supercritical carbon dioxide (SC-CO)₂) Blow-drying, and after timing for 1h, obtaining 95.9mg of microcapsule inclusion compound from the kettle. The medicine-loading rate of naringenin in the naringenin-HP-beta-CD binary microcapsule inclusion compound prepared in the embodiment is 70.13%, and the cumulative dissolution rate is 13.04% in 130 min.

The dissolution rate of the naringenin-HP-beta-CD binary microcapsule inclusion compound is tested, and the test result shows that: at any moment, the dissolution rate of the naringenin-HP-beta-CD binary microcapsule inclusion compound is far higher than that of a physical mixture of naringenin and naringenin/HP-beta-CD, and the cumulative dissolution rate reaches 13.04% at 130 min.

Example 10

Weighing 100.2mg of quercetin and 150.1mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD), adding all the quercetin into a beaker, adding 40mL of anhydrous methanol, stirring uniformly, and dispersing by using ultrasonic to obtain a uniform and transparent solution. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring and starting to release the solvent after timing is finished. Observing quadrant change in the window, releasing solvent completely, and starting to use supercritical carbon dioxide (SC-CO)₂) Blow-drying, and obtaining 55.0mg of microcapsule inclusion compound from the kettle after timing for 1 h. The shaddock in the quercetin HP-beta-CD binary microcapsule inclusion compound prepared in the embodiment is detectedThe drug loading rate of the skin extract is 15.50%, and the cumulative dissolution rate after 130min can reach 54.57% (see figure 16)

FIG. 8 is a SEM image of the binary quercetin-HP-beta-CD microcapsule prepared in this example. The figure shows that the honeycomb porous structure formed by winding the micro-fibers has a large specific surface area.

FIG. 10 shows a particle size analysis chart of Quercetin-HP-beta-CD binary clathrate. Wherein, D10 is 5.51 μm; d50 ═ 24.48 μm; d90 ═ 33.93 μm; d99 ═ 45.13 μm; dav 23.67 μm. D10 represents particles having a particle size less than (or greater than) that of 10%. The meanings of D50, D90 and D99 are analogized.

Example 11

100.5mg of naringenin and 150.0mg of hydroxypropyl methylcellulose (HPMC) are weighed and added into a beaker, 40mL of anhydrous methanol is added, and the mixture is dispersed into a uniform and transparent solution by using ultrasonic after being stirred uniformly. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring and starting to release the solvent after timing is finished. Observing quadrant change in the window, completely releasing the solvent, starting to blow and dry with supercritical carbon dioxide, timing for 1h, and obtaining 89.6mg of microcapsule inclusion compound from the kettle. The medicine-loading rate of the naringenin-HPMC microcapsule inclusion compound prepared in the embodiment is 16.8834%.

Example 12

Weighing 100.4mg of quercetin and 150.1mg of hydroxypropyl methylcellulose (HPMC), completely adding into a beaker, adding 40mL of anhydrous methanol, stirring uniformly, and dispersing by using ultrasonic to obtain a uniform and transparent solution. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring at 500rpm for timing for 30min after the temperature and the pressure reach set values, stopping stirring after timing is finished, and starting to release the solvent. Observing quadrant change in the window, releasing solvent completely, and starting to use supercritical carbon dioxide (SC-CO)₂) Blow-drying, and obtaining 108.4mg of microcapsule inclusion compound from the kettle after timing 1 h. The drug loading rate of the quercetin-HPMC binary microcapsule inclusion compound prepared in this example was 47.41%.

As shown in fig. 9, the cumulative dissolution rate (tested by the conventional method) of the quercetin-HPMC binary microcapsule inclusion compound, quercetin, and the physical mixture of quercetin and HPMC in the present example is between 0-140 min. As can be seen from the figure, the test results show that: at any moment, the dissolution rate of the quercetin-HPMC binary microcapsule inclusion compound prepared by the invention is far higher than that of a physical mixture of quercetin and quercetin/HPMC, and the cumulative dissolution rate can reach 53.95% in 140 min.

Example 13

Weighing luteolin 100.2mg and hydroxypropyl methylcellulose (HPMC)150.4mg, adding into a beaker, adding 40mL anhydrous methanol, stirring well, and dispersing with ultrasound to obtain uniform and transparent solution. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm, starting timing for 30min, stopping stirring after timing is finished, and starting to release the solvent. Observing quadrant change in the window, completely releasing the solvent, drying with supercritical carbon dioxide, and timing for 1 hr to obtain microcapsule clathrate (103.5 mg). The drug loading rate of the luteolin-HPMC binary microcapsule inclusion compound prepared by the embodiment is 12.0134%.

Example 14

Weighing 100.1mg of luteolin, 150.7mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD) and 100.3mg of polyvinylpyrrolidone (PVP K-30), adding into a beaker, adding 40mL of anhydrous methanol, stirring well, and dispersing with ultrasound to obtain a uniform and transparent solution. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, stopping stirring after timing is finished, and starting to release the solvent. Observing quadrant change in the window, completely releasing the solvent, drying with supercritical carbon dioxide, and collecting 49.2mg microcapsule clathrate from the kettle after 1 hr. The luteolin-HP-beta-CD-PVP ternary microcapsule inclusion compound prepared by the embodiment has the drug loading rate of 14.4443%, and the maximum 130-min cumulative dissolution rate of 59.3641%.

Example 15

100.7mg of apigenin and 150.2mg of hydroxypropyl methylcellulose (HPMC) are weighed and added into a beaker, 20mL of absolute ethyl alcohol and 20mL of dichloromethane are added, and after uniform stirring, the mixture is dispersed into a uniform and transparent solution by using ultrasound. Adding the solution into a supercritical visible kettle, closing the kettle body, setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach a set value, setting 500rpm, starting timing for 30min, stopping stirring after timing is finished, and starting to release the solvent. Observing quadrant change in the window, completely releasing the solvent, drying with supercritical carbon dioxide, and timing for 1 hr to obtain 161.6mg microcapsule clathrate. The drug loading rate of the apigenin-HPMC binary microcapsule inclusion compound prepared by the embodiment is 29.6792%, and the maximum 130-min cumulative dissolution rate is 39.2159%.

Example 16

200.1mg of apigenin and 300.1mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD) are weighed, all the apigenin and the 2- (hydroxypropyl) -beta-cyclodextrin are added into a beaker, 20mL of absolute ethyl alcohol and 20mL of dichloromethane are added, and the mixture is dispersed into a uniform and transparent solution by using ultrasound after being stirred uniformly. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring and starting to release the solvent after timing is finished. Observing quadrant change in a window, completely releasing the solvent, drying by using supercritical carbon dioxide, and after timing for 1h, obtaining 348.5mg of the microcapsule inclusion compound from the kettle to obtain the binary microcapsule inclusion compound of apigenin-HP-beta-CD.

Example 17

Weighing 200.0mg of apigenin, 300.1mg of 2- (hydroxypropyl) -beta-cyclodextrin (HP-beta-CD) and 200.2mg of polyvinylpyrrolidone (PVP K-30); the mixture was added to a beaker, 40mL of absolute ethanol and 20mL of methylene chloride were added thereto, and the mixture was uniformly stirred and then dispersed by ultrasonic waves to obtain a uniform and transparent solution. Adding the solution into a supercritical visible kettle, and closing the kettle body. Setting the temperature at 45 ℃ and the pressure at 14MPa, starting stirring after the temperature and the pressure reach set values, setting 500rpm, starting timing for 30min, and stopping stirring and starting to release the solvent after timing is finished. Observing quadrant change in a window, completely releasing the solvent, drying by using supercritical carbon dioxide, and after timing for 1h, obtaining 465.6mg of the microcapsule inclusion compound from the kettle to obtain the apigenin-HP-beta-CD-PVP ternary microcapsule inclusion compound.

The microcapsule formulations prepared in examples 1-17 were loaded, and the cumulative maximum dissolution at 130min (140 min in part) and average particle size statistics are given in the following table:

from examples 1-3, 8, 10, and 12, it is known that when two carriers of HPMC and PVP are used together in the preparation of the quercetin microcapsule inclusion compound, and the mass ratio of the raw material to the carrier is 1:2.5, the drug-loading rate of the prepared quercetin microcapsule inclusion compound is significantly higher. However, when cyclodextrin and quercetin were used to prepare binary inclusion compounds (as in example 10), although the drug loading was only 15.50%, the final release at 130min was much higher than that of the other inclusion compounds. The pH responsiveness thereof was proved to be superior to the other examples. Therefore, for quercetin, HP-beta-CD is used as a carrier for preparing the microcapsule inclusion compound, so that the carrier has advantages over other carriers.

Comparative examples 4, 13, 14; in the case of luteolin, although the drug loading of example 14 is lower than that of example 4, the final cumulative dissolution release after 130min can reach 59.36%, so that the HP-beta-CD-PVP dual carrier is considered to have better response to pH than the HPMC-PVP dual carrier, and the drug can be well and qualitatively released.

Comparative examples 5, 7, 9, 11; for naringenin, although the drug loading rate of example 7 is lower than that of example 9, the naringenin has more excellent pH responsiveness, and the cumulative dissolution rate at 130min can reach 82.83%. In all examples, the cumulative dissolution of naringenin-HPMC-PVP ternary microencapsulation inclusion complex prepared in example 7 was the highest, indicating that the HPMC-PVP binary vehicle is very suitable for preparing naringenin inclusion complex.

In comparison to examples 6, 15-17, example 15 was able to achieve better pH response for apigenin and its drug loading was 29.68% higher than that of the other examples. As can be seen, the apigenin-HPMC binary microcapsule inclusion compound is superior to the apigenin-HPMC-PVP (or HP-beta-CD-PVP) ternary microcapsule inclusion compound, can achieve a good dissolution effect, and further improves the bioavailability.

After the carrier is selected, good inclusion can be achieved when the mass ratio of the medicine to the carrier is 1: 2.5. In some embodiments, multiple carriers are coated with the same drug and stacked to form a microcapsule.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.