Long-acting pellet suitable for stomach absorption and preparation method thereof

Abstract

The invention discloses a long-acting pellet suitable for gastric absorption and a preparation method of the long-acting pellet. The long-acting pellet is prepared from the following components in parts by mass: 0.01-1 part of a medicine, 1-2.5 parts of a filler, 0.2-0.7 part of a modified pore-foaming agent mixture, 1-2.5 parts of an improved adhesive mixture and 0.1-0.7 part of an improved sonokinetic response agent. Wherein the modified pore-foaming agent mixture is prepared from a modified pore-foaming agent and glycosylated saponin, the modified pore-foaming agent is prepared by mixing a pore-foaming agent and seleno-amino acid according to a mass ratio of 4: 1, the improved adhesive is mixed with the adhesive through BHP-SH, and ionic crosslinking is performed by using CaCl2 to form a covalent ion dual network, so that the stomach adhesiveness is enhanced; the sonokinetic response agent is improved by the coated poria cocos extract, and the long-acting pellet is prepared by adopting a wet granulation method or a fluidized bed method, has the characteristics of slow release, long acting, targeted stomach absorption and responsive drug release, and is suitable for treating stomach diseases such as gastritis.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical technology, specifically relating to a long-acting microsphere suitable for gastric absorption and its preparation method. Background Technology

[0002] Microcapsules are spherical or near-spherical solid dosage forms with a diameter of 0.5 to 1.0 mm. They can also be encapsulated, compressed into tablets, or made into other formulations. Microcapsules are a multi-unit oral dosage form, and the amount of drug administered at one time usually consists of dozens to hundreds of microcapsules. They can be made into different types of microcapsules such as sustained-release, enteric-coated, etc.

[0003] Microparticles have unique characteristics, including: aesthetically pleasing appearance, good flowability, wide drug loading range, can be used to make sustained-release and controlled-release formulations, stable drug release, high bioavailability, avoidance of incompatibilities between multiple drugs, reduction of drug irritation to the digestive tract, and masking of unpleasant tastes of certain drugs.

[0004] Treatment of stomach diseases (gastritis, gastric ulcer, gastroesophageal reflux disease, etc.) usually requires drugs to exert their effects locally in the stomach. However, traditional microcapsules, after oral administration, are widely distributed in the gastrointestinal tract and cannot be localized to the stomach to exert a long-term effect.

[0005] Compared to floating gastric microspheres, gastric adhesive microspheres offer several advantages, particularly in that their gastric retention time is not affected by body position. The adhesiveness of gastric adhesive microspheres primarily relies on bioadhesives or mucosal adhesives, which enable drug formulations to firmly adhere to the gastric mucosa, thereby prolonging the drug's residence time in the stomach.

[0006] Currently, traditional gastric adhesive microspheres still face several unresolved issues. One of the most prominent problems is the poor adhesion between gastrointestinal adhesive microspheres and the stomach wall. One of the main reasons for this is the barrier effect of biological tissues and the limited penetration and adhesion of bioadhesives by the moist gastric mucosa, which is difficult to overcome for bioadhesives based on physical interactions. Bioadhesives that can adapt to the complex human environment have always been an important direction for scientific research. Although some chemical strategies have achieved high adhesion, the reagents contained in the adhesives and the chemical reactions with biological tissues can generate new problems, such as interference with drug delivery, residues after degradation, and inflammatory reactions that may be triggered by reaction byproducts. Summary of the Invention

[0007] To address the issue of poor adhesion between gastric adhesive microspheres and the gastric wall, this application provides a long-acting microsphere suitable for gastric absorption and its preparation method. Through the synergistic effect of three functional modification mixtures (porogen, adhesive, and acoustic responsive agent), a microsphere system with long-acting gastric absorption characteristics is constructed: The modified porogen mixture consists of a 1:1 compounding of a selenoamino acid-modified porogen and a glycosylated saponin (β-glucosidase hydrolysis product) to form a mixed micelle structure, regulating the drug release rate; the modified adhesive mixture... 2+ Ionic crosslinking and thiol oxidative covalent bonds (-SS-) form a dual network, enhancing gastric mucosal adhesion (based on the combination of winter melon extract BHP-SH and conventional adhesives); modified acoustic response agents: carbonates combined with ethyl cellulose-coated Poria cocos extract achieve ultrasound-triggered response release.

[0008] This application provides, on the one hand, long-acting microspheres suitable for gastric absorption, wherein the long-acting microspheres are made of the following components in the following mass ratio: 0.01-1 parts of drug, 1-2.5 parts of filler, 0.2-0.7 parts of modified porogen mixture, 1-2.5 parts of modified binder mixture, and 0.1-0.7 parts of modified acoustic response agent, wherein the modified porogen mixture is prepared by mixing the modified porogen with glycosylated saponin at a mass ratio of 1:1; the modified binder mixture is formed by ionic cross-linking of the binder mixture with CaCl2 to form a covalent ionic double network; and the modified acoustic response agent is prepared by mixing the acoustic response agent with coated Poria cocos extract at a mass ratio of 9:1.

[0009] Preferably, the drug is a gastritis drug, wherein the gastritis drug is any one of omeprazole, rabeprazole, clarithromycin, and amoxicillin; and the filler is any one of microcrystalline cellulose, lactose, and pregelatinized starch.

[0010] Preferably, the modified porogen is a mixture of a porogen and a selenoamino acid at a mass ratio of 4:1. The porogen is any one of polyethylene glycol 2000, sodium dodecyl sulfate, and polyethylene glycol 3000. The glycosylated saponin is a product of β-glycosidase hydrolysis of Fritillaria cirrhosa saponin. The preparation method of the modified porogen mixture is as follows: (1) Preparation of glycosylated saponins: 1-5 mg / mL of substrate fritillaria saponin, 5-10 eq of p-nitrophenyl-β-D-glucoside, β-glucosidase (10-20 U / mg substrate), and tert-butanol-water (4:1 v / v) were dissolved in sodium acetate buffer and incubated in a water bath at 50°C in the dark for 2 h. TLC monitoring was performed, and an equal volume of methanol was added. The mixture was in an ice bath for 10 min, at 10000 rpm for 10 min, at 4°C. The supernatant was collected and concentrated. Ethyl acetate was added for extraction to obtain crude glycosylated saponins. The purified glycosylated saponins were obtained by silica gel column purification. (2) Preparation of modified porogen: any one of the porogens polyethylene glycol 2000, sodium dodecyl sulfate, and polyethylene glycol 3000 is mixed with selenoamino acid at a mass ratio of 4:1 and dissolved in deionized water until completely dissolved to obtain the modified porogen. (3) Mixing: The modified porogen and glycosylated saponin were dissolved in 30% ethanol solution at a mass ratio of 1:1 and dissolved by ultrasonic assistance to form a clear solution; the parameters were 40 kHz, 30 min, and 25 ℃. (4) Self-assembly of mixed micelles: The solution was kept under magnetic stirring (500 rpm, 25°C) for 2 h to allow the modified porogen to combine with the hydrophobic portion of the glycosylated saponin to spontaneously form mixed micelles; (5) Purification and drying: The mixture was placed in a dialysis bag and dialyzed with deionized water for 24 hours (water was changed every 4 hours) to remove unbound modified porogens and free glycosylated saponins. The mixture was then freeze-dried (pre-frozen at -80°C and freeze-dried for 24 hours) to obtain a solid mixed micelle powder modified porogen mixture. (6) Characterization: The micelles were observed to be spherical or nearly spherical by radio electron microscopy.

[0011] Preferably, the adhesive mixture is prepared by mixing any one of the adhesives polyacrylic acid, chitosan, carrageenan, sodium alginate, carbomer, and hydroxypropyl methylcellulose with BHP-SH at a mass ratio of 4:1. The BHP-SH is obtained by grafting winter melon extract (BHP) with L-cysteine.

[0012] Preferably, the method for preparing the modified adhesive mixture and the adhesive mixture is as follows: (1) Preparation of winter melon extract: 1) Grinding and homogenization: Chop, homogenize or ultrafine the winter melon pulp to break down cell walls and release more pectin and polysaccharides; 2) Heating extraction: High-temperature cooking (80~100℃) promotes the dissolution of pectin and polysaccharides and enhances hydration; a weak gel may form after cooling. 3) Freeze-thaw cycle: The winter melon pulp is repeatedly frozen (-20℃) and thawed (room temperature) to destroy the cell structure. The formation of ice crystals makes the polysaccharide network looser and increases the porosity after absorbing water. 4) Compound enzymatic hydrolysis: Add pectinase (0.5% w / v) + cellulase (0.2% w / v), pH 4.5, hydrolyze at 50℃ for 2 h, inactivate enzyme at 90℃ for 10 min, and then dialysis (molecular weight cutoff 10 kDa) to purify BHP.

[0013] (2) Preparation of adhesive mixture: 1) BHP pretreatment: Dissolve the purified BHP in 0.1M PBS buffer (pH=6.0) to prepare a 5% (w / v) solution. Stir overnight at 4°C to completely dissolve the BHP. Centrifuge (8000 rpm, 15 min) to remove insoluble matter. 2) Carboxyl activation: Add EDC·HCl (2 times the molar amount of BHP carboxyl group) and NHS (1.2 times the molar amount of EDC) to step (1), and stir the reaction at 25°C in the dark for 2 hours under nitrogen protection, so that the pH is maintained at 6.0±0.2; 3) Thiohydration reaction: Add L-cysteine ​​(1.5 times the molar amount of BHP carboxyl group), react in a 40℃ water bath under nitrogen protection for 6 hours, and maintain the pH at 5.0±0.2; 4) Purification: Transfer the reaction solution from step (3) to a dialysis bag and dialyze it: 0.1M NaCl solution containing 1mM EDTA (24h), deionized water (48h, water changed 6 times), freeze-dry to obtain BHP-SH white powder; 5) Dissolve any one of the adhesives polyacrylic acid, chitosan, carrageenan, sodium alginate, carbomer, and hydroxypropyl methylcellulose in deionized water at a mass ratio of 4:1 with BHP-SH in step (4), mix, and dry to obtain an adhesive mixture.

[0014] (3) Preparation of modified adhesive mixtures: 1) Ca 2+ Network formation: Dissolve the adhesive mixture in 5 mL of deionized water, add 1 mL of 2% CaCl2 solution, and gently stir (200 rpm) for 30 min under nitrogen protection, avoiding bubble formation. Let stand for 1 h to allow the CaCl2 to form. 2+ Uniform diffusion crosslinking; 2) Covalent crosslinking: The mixture was transferred to a silica gel petri dish and placed in a 37°C incubator for 6 hours to promote the spontaneous oxidation of -SH to form -SS-. After gel formation, unreacted Ca was removed by rinsing with deionized water. 2+ ; 3) Drying: freeze-dry (pre-freeze at -80℃, vacuum dry for 24h) to obtain a porous sponge-like modified adhesive mixture.

[0015] Preferably, the acoustic response agent is any one of calcium carbonate, sodium carbonate, or sodium bicarbonate, and the coated Poria cocos extract is prepared by mixing Poria cocos extract and ethyl cellulose at a mass ratio of 3:1.

[0016] Preferably, the method for preparing the modified acoustic response agent is as follows: (1) Dissolving ethyl cellulose: Mix ethyl cellulose with anhydrous ethanol at a ratio of 1:10 (w / v) and stir magnetically for 2 hours until completely dissolved to obtain EC ethanol solution (10% w / v). (2) Coating process: Place the Poria cocos extract (300g) in a fluidized bed coating machine, preheat the material to 40°C, spray in EC ethanol solution (100g EC dissolved in 1L ethanol), inlet air temperature 50°C, atomization pressure 0.8 bar, flow rate 5mL / min, after coating, dry at 40°C for 2h, pass through an 80-mesh sieve to obtain coated Poria cocos extract; (3) Improved acoustic response agent: Take 900g of acoustic response agent and 100g of coated Poria cocos extract in a 9:1 mass ratio and place them in a three-dimensional mixer. Mix for 30min (20rpm) to homogenize. Dry under vacuum at 40℃ for 4h to obtain the improved acoustic response agent.

[0017] Another aspect of this application provides a method for preparing long-acting microspheres suitable for gastric absorption, characterized in that the preparation method is a wet particle preparation method and a fluidized bed preparation method.

[0018] Preferably, the wet granule preparation method comprises the following steps: (1) Grind the drug and filler into powder separately, pass them through a 60-150 mesh sieve, and mix them thoroughly. (2) The drug, filler powder, pore-forming agent mixture, modified adhesive mixture, and modified acoustic response agent described in step (1) are mixed in the mass ratio described in claim 1; (3) Spray in a 5%~50% ethanol solution to make a soft material; (4) The soft material is made into wet granules through a sieve with a pore size of 2~3mm; (5) The wet granules are placed in a drying equipment for drying to remove moisture; (6) After drying, the granules are sized by passing them through a 1-2 mm sieve and then sealed and packaged.

[0019] Preferably, the fluidized bed preparation method comprises the following steps: (1) Grind the drug and filler into powder separately, pass them through a 60-150 mesh sieve, and mix them thoroughly. (2) The drug, filler powder, pore-forming agent mixture, modified adhesive mixture, and modified acoustic response agent described in step (1) are mixed in the mass ratio described in claim 1; (3) Place it in a fluidized bed to complete the granulation-drying-sizing process, and then seal and package it.

[0020] The beneficial effects of the embodiments in this application are as follows: (1) A modified porogen mixture (porogen + glycosylated saponin) is used to improve the controllability of drug release. Glycosylated saponin is modified by enzymatic method to enhance biocompatibility, improve hydrophilicity, and optimize swelling behavior, so that the drug is slowly released in gastric juice, interacts with gastric mucus glycoproteins, enhances mucosal adhesion, and prolongs retention time. The porogen can regulate the porosity of microspheres and affect the drug diffusion rate. Selenized amino acids are modified to enhance antioxidant properties and protect the drug from gastric acid degradation. (2) The adhesive provides basic adhesion, electrostatically adsorbs onto the gastric mucosa, and improves the adhesive through Ca... 2+ Ionic crosslinking and -SS-covalent bonds form a dual network structure, which significantly prolongs the gastric retention time (experiments show that the adhesion force is increased by 3.2 times). (3) The acoustic response agent generates CO2 microbubbles in the gastric acid environment, which promotes drug diffusion. It combines with coated Poria cocos extract to form a modified acoustic response agent, which delays the acid reaction of carbonate and achieves ultrasound-triggered release. (4) The microspheres adhere firmly to the stomach wall: The modified acoustic response agent promotes the generation of a large number of microbubbles near the microspheres, which are quickly applied to the stomach wall. At the same time, the acoustic effect produced by the modified acoustic effect not only helps to overcome the mucosal barrier, but also enhances the adhesion between the adhesive and the stomach wall, avoiding the problem of short retention time in the stomach due to insufficient adhesion, and ensuring the effective release of the drug in the stomach. (5) Enhance the therapeutic effect of gastric diseases: Microparticles can form a large surface area in the stomach, thereby more effectively contacting the gastric mucosa, increasing the local concentration of the drug, and enhancing the therapeutic effect of the drug. This is of great clinical significance for the treatment of gastric diseases such as gastric ulcers and gastritis. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the in vitro release data of Example 1 of this application.

[0022] Figure 2 This is a schematic diagram of the in vitro release data for Example 1 and Example 2 of this application.

[0023] Figure 3 This is a schematic diagram of the in vitro release data for Example 1, Example 3 of this application.

[0024] Figure 4 This is a schematic diagram of the in vitro release data for Example 1, Example 4 of this application.

[0025] Figure 5 This is a schematic diagram of the in vitro release data for Example 1, Example 5 of this application.

[0026] Figure 6 This is a schematic diagram of the in vitro release data for Example 1, Example 6 of this application.

[0027] Figure 7 This is a schematic diagram of the in vitro release data of the comparative example 1 of this application.

[0028] Figure 8 This is a schematic diagram of the adhesion force data for Test Example 2 of this application. Detailed Implementation

[0029] The technical solutions of this application will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0030] The reagents and equipment used in the embodiments of this disclosure are all conventional and commercially available.

[0031] Preparation Example 1 Preparation of modified porogen mixture (1) Extraction of Fritillaria cirrhosa saponins: 1) Preparation before extraction: Select dry, mold-free Fritillaria cirrhosa medicinal material, remove impurities, crush into coarse powder and pass through a 30-mesh sieve to increase the contact area and improve the extraction efficiency to obtain Fritillaria cirrhosa coarse powder. 2) Extraction: Take 100g of crude Fritillaria cirrhosa powder, add 1000mL of deionized water, boil three times, filter and extract each time, each boiling time is 1.5h, and combine the filtrates. 3) Concentration: The combined filtrate from step (2) is concentrated at 60°C and -0.08 MPa until it becomes a thick, viscous paste. Add 95% ethanol and monitor with an alcohol meter until the alcohol content reaches 60%. Stop adding ethanol and continue stirring for 30 minutes to allow the components to fully contact the ethanol. Transfer to a clean container, seal, and let stand at 20°C for 20 hours. 4) Filtration and recovery: The supernatant and precipitated impurities are separated by filter paper. The supernatant is used to recover ethanol at 60°C using a rotary evaporator until there is no obvious ethanol odor, thus obtaining Fritillaria cirrhosa saponin extract. 5) Acid hydrolysis: Dissolve the Fritillaria cirrhosa saponin extract in deionized water at 50°C (solid-to-liquid ratio 1:10), then add 2 mol / L hydrochloric acid to adjust the pH of the solution to 1-2; transfer to a reflux apparatus and hydrolyze under boiling water bath conditions at 100°C for 3 hours. After hydrolysis, cool the solution to room temperature and extract 5 times with ethyl acetate at a volume ratio of 1:1 to the hydrolysate. Transfer the saponins produced by hydrolysis to the organic phase, combine the organic phases, and concentrate using a rotary evaporator to obtain crude Fritillaria cirrhosa saponins. After purification, obtain high-purity Fritillaria cirrhosa saponins.

[0032] (2) Preparation of glycosylated saponins: 1) System preparation: Measure 6 mL of sodium acetate buffer (pH 5.0), add 2.4 mL of tert-butanol + 0.6 mL of water (to prepare a 4:1 v / v mixed solvent), stir well, and use it as the basic reaction solvent system; 2) Enzyme-initiated reaction: Weigh 2 mg of Fritillaria cirrhosa saponin and add it to the above mixed solvent. Then weigh 9.48 mg of p-nitrophenyl-β-D-glucoside and add it to the system. Stir until dissolved, add 20 U of β-glucoside enzyme, and bring the system to a final volume of 10 mL. 3) Enzymatic glycosylation reaction: Transfer the reaction system to a stoppered test tube, place it in a 50℃ constant temperature water bath in the dark, and react for 2 hours. Gently shake the tube every 15 minutes during the reaction to make the reaction more uniform. At 0.5h, 1h, and 2h, take 10μL of the reaction solution and spot it onto a TLC plate. Develop the plate with the developing solvent, spray with the color reagent, and heat to develop the color (bake at 105℃ for 3 minutes). Observe: The substrate (Fritillaria cirrhosaponin) spots gradually fade over time, while the product (glycosylated saponin) spots gradually darken, indicating that the reaction is proceeding. 4) Termination of reaction and separation: Add 10 mL of methanol, incubate on ice for 10 min, at 10000 rpm for 10 min, at 4℃. Transfer the supernatant to a rotary evaporator flask and concentrate at 50℃ and -0.08 MPa to 1 mL. Add 3 mL of ethyl acetate for extraction for 15 min (glycosylated saponins are readily soluble in ethyl acetate, and most impurities remain in the aqueous phase). Transfer to a separatory funnel, let stand for 10 min to separate the layers, discard the lower aqueous phase, and transfer the upper ethyl acetate phase to a new flask. Concentrate under reduced pressure (under the same conditions) to obtain the crude glycosylated saponin extract. 5) Silica gel column purification: The purified glycosylated saponins were obtained by silica gel column purification.

[0033] (3) Preparation of modified porogens: Take 4g of pore-forming agent and 1g of selenoamino acid in a clean and dry beaker, add deionized water to 30mL, heat to 35℃ to dissolve, and spray dry. The inlet air temperature is 150℃, the outlet air temperature is 80℃, and the peristaltic pump speed is 10mL / min. After drying is completed, turn off the equipment, take out the dried particles from the collection device, and sieve through a 100-mesh sieve to remove fine powder and clumps, so as to obtain uniform spray-dried particles.

[0034] (4) Preparation of modified porogen mixture: The modified porogen and glycosylated saponin were dissolved in 30% ethanol solution at a mass ratio of 1:1 and dissolved with ultrasonic assistance to form a clear solution; the parameters were 40 kHz, 30 min, 25 °C; the solution was maintained under magnetic stirring (500 rpm, 25 °C) for 2 h to allow the modified porogen and the hydrophobic part of the glycosylated saponin to combine spontaneously to form mixed micelles. The mixture was put into a dialysis bag and dialyzed with deionized water for 24 h (water was changed every 4 h) to remove unbound modified porogen and free glycosylated saponin. The mixture was freeze-dried (pre-frozen at -80 °C, freeze-dried for 24 h) to obtain solid mixed micelle powder and modified porogen mixture. The micelle morphology was observed to be spherical or nearly spherical by radio electron microscopy.

[0035] It should be noted that the pore-forming agent is any one of polyethylene glycol 2000, sodium dodecyl sulfate, and polyethylene glycol 3000, ultimately yielding a mixture of modified polyethylene glycol 2000, a mixture of modified sodium dodecyl sulfate, and a mixture of modified polyethylene glycol 3000.

[0036] Preparation Example 2 Preparation of modified adhesive mixtures (1) Preparation of winter melon extract: 1) Grinding and homogenization: Chop, homogenize or ultrafine the winter melon pulp to break down cell walls and release more pectin and polysaccharides; 2) Heating extraction: High-temperature cooking at 90℃ promotes the dissolution of pectin and polysaccharides and enhances hydration; a weak gel may form after cooling. 3) Freeze-thaw cycle: The winter melon pulp is repeatedly frozen (-20℃) and thawed (room temperature) to destroy the cell structure. The formation of ice crystals makes the polysaccharide network looser and increases the porosity after absorbing water. 4) Compound enzymatic hydrolysis: Add pectinase (0.5% w / v) + cellulase (0.2% w / v), pH 4.5, hydrolyze at 50℃ for 2 h, inactivate enzyme at 90℃ for 10 min, and then dialysis (molecular weight cutoff 10 kDa) to purify BHP.

[0037] (2) Preparation of adhesive mixture: 1) BHP pretreatment: Dissolve the purified BHP in 0.1M PBS buffer (pH=6.0) to prepare a 5% (w / v) solution. Stir overnight at 4℃ to completely dissolve the BHP. Centrifuge (8000 rpm, 15 min) to remove insoluble matter. 2) Carboxyl activation: Add EDC·HCl (2 times the molar amount of BHP carboxyl group) and NHS (1.2 times the molar amount of EDC) to step (1), and stir the reaction at 25°C in the dark for 2 hours under nitrogen protection, so that the pH is maintained at 6.0±0.2; 3) Thiohydration reaction: Add L-cysteine ​​(1.5 times the molar amount of BHP carboxyl group), react in a 40℃ water bath under nitrogen protection for 6 hours, and maintain the pH at 5.0±0.2; 4) Purification: Transfer the reaction solution from step (3) to a dialysis bag and dialyze it: 0.1M NaCl solution containing 1mM EDTA (24h), deionized water (48h, water changed 6 times), freeze-dry to obtain BHP-SH white powder; 5) Dissolve the adhesive and BHP-SH from step (4) in deionized water at a mass ratio of 4:1, mix and dry to obtain the adhesive mixture.

[0038] (3) Preparation of modified adhesive mixtures: 1) Ca 2+ Network formation: Dissolve the adhesive mixture in 5 mL of deionized water, add 1 mL of 2% CaCl2 solution, and gently stir (200 rpm) for 30 min under nitrogen protection, avoiding bubble formation. Let stand for 1 h to allow the CaCl2 to form a network. 2+ Uniform diffusion crosslinking; 2) Covalent crosslinking: The mixture was transferred to a silica gel petri dish and placed in a 37°C incubator for 6 hours to promote the spontaneous oxidation of -SH to form -SS-. After gel formation, unreacted Ca was removed by rinsing with deionized water. 2+ ; 3) Drying: freeze-dry (pre-freeze at -80℃, vacuum dry for 24h) to obtain a porous sponge-like modified adhesive mixture.

[0039] It should be noted that the adhesive is any one of polyacrylic acid, chitosan, carrageenan, sodium alginate, carbomer, and hydroxypropyl methylcellulose, ultimately yielding a modified polyacrylic acid mixture, a modified chitosan mixture, a modified carrageenan mixture, a modified sodium alginate mixture, a modified carbomer mixture, and a modified hydroxypropyl methylcellulose mixture.

[0040] Preparation Example 3 Preparation of improved acoustic response agents (1) Dissolving ethyl cellulose: Dissolve 1g of ethyl cellulose in 10mL of anhydrous ethanol and stir magnetically at 200rpm for 2h until completely dissolved to obtain EC ethanol solution; (2) Coating process: Place 300g of Poria cocos extract in a fluidized bed coating machine and preheat to 40°C to ensure material flowability; spray in EC ethanol solution (100g EC dissolved in 1L ethanol), inlet air temperature 50°C, atomization pressure 0.8 bar, flow rate 5mL / min. After coating, dry at 40°C for 2 hours and pass through an 80-mesh sieve to obtain coated Poria cocos extract. (3) Improved acoustic response agent: Take 900g of acoustic response agent and 100g of coated Poria cocos extract in a 9:1 mass ratio and place them in a three-dimensional mixer. Mix for 30min (20rpm) to homogenize. Dry at 40℃ under vacuum of -0.08MPa for 4h to obtain the improved acoustic response agent.

[0041] It should be noted that the acoustic response agent is any one of calcium carbonate, sodium carbonate, or sodium bicarbonate, ultimately yielding modified calcium carbonate, modified sodium carbonate, or modified sodium bicarbonate.

[0042] Example 1 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: After crushing the filler, pass it through a 100-mesh sieve. The relative humidity of the environment is ≤40% (to prevent the raw material from absorbing moisture). The modified pore-forming agent mixture is prepared in the same way as in Preparation Example 1, the modified adhesive mixture is prepared in the same way as in Preparation Example 2, and the modified acoustic response agent is prepared in the same way as in Preparation Example 3. (2) Calculation of addition amount: The preparation was carried out according to the following mass ratio of each component: 0.5 parts barium sulfate drug, 1.5 parts filler, 0.5 parts modified pore-forming agent mixture, 1.5 parts modified adhesive mixture, and 0.5 parts modified acoustic response agent. The calculation formula is as follows: The specific addition amounts are shown in Table 1: Table 1: Amounts of each component added in Example 1

[0043] (3) Preparation: Wet granule preparation: Put the above components into the mixer according to the data in the table, first premix at a low speed of 15 rpm for 5 min, then adjust to a medium speed of 25 rpm for 15 min to ensure that the materials are mixed evenly, and pass through an 80-mesh sieve to remove lumps or coarse particles; spray in a 10% ethanol solution at a spray rate of 8 mL / min, and at the same time turn on the mixer to stir at a speed of 15 rpm to make a soft material; pass the soft material through a sieve with a pore size of 2~3 mm to make wet granules; place the wet granules in a vacuum drying oven to dry at a temperature of 50℃ and a vacuum degree of ≤-0.08MPa for 3 hours to remove moisture, and then pass the dried granules through a 2 mm sieve to granulate and seal them for packaging.

[0044] Example 2 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: Same as in Example 1; (2) Calculation of addition amount: Same as in Example 1, the specific addition amount data is shown in Table 2: Table 2: Amounts of each component added in Example 2

[0045] (3) Preparation: Fluidized bed preparation: Add the above components to the mixer according to the data in the table, premix at a low speed of 15 rpm for 5 min, then adjust to a medium speed of 25 rpm for 15 min to ensure uniform mixing. Remove lumps or coarse particles by passing through an 80-mesh sieve, spray in a 10% ethanol solution for binding, and place in a fluidized bed; the fluidized bed granulation parameters are set as follows: inlet air temperature 55℃, atomization pressure 1.5 bar, spray rate 3-5 mL / min, fluidizing air volume 30 m³ / min. 3 / h, material temperature 30~35℃, add premixed powder to fluidized bed hopper, initial fluidization (air volume 30m³ / h) for 5min preheating, turn on spray, spray liquid evenly with concentric spray gun, spray distance 15~20cm, first at low speed (3mL / min) to wet the surface, then adjust to 5mL / min to the endpoint, after spraying stops, maintain fluidization and drying for 15min, wait for the particles to cool down to 25℃, granulate through 2mm sieve, seal and package to obtain the product.

[0046] Example 3 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: Same as in Example 1; (2) Calculation of addition amount: Same as in Example 1, the specific addition amount data is shown in Table 3: Table 3: Amounts of each component added in Example 3

[0047] (3) Preparation: Same as in Example 1.

[0048] Example 4 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: Same as in Example 1; (2) Calculation of addition amount: Same as in Example 1, the specific addition amount data is shown in Table 4: Table 4: Amounts of each component added in Example 4

[0049] (3) Preparation: Same as in Example 2.

[0050] Example 5 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: Same as in Example 1; (2) Calculation of addition amount: Same as in Example 1, the specific addition amount data is shown in Table 5: Table 5: Amounts of each component added in Example 5

[0051] (3) Preparation: Same as in Example 1.

[0052] Example 6 Barium sulfate was used as a model drug to prepare 270mg retention tablets. (1) Raw material preparation: Same as in Example 1; (2) Calculation of addition amount: Same as in Example 1, the specific addition amount data is shown in Table 6: Table 6: Amounts of each component added in Example 6

[0053] (3) Preparation: Same as in Example 2.

[0054] Comparative Examples 1-6: Barium sulfate was used as a model drug to prepare 270mg retention tablets. The components of the comparative example are shown in Table 7. The preparation method parameters and component addition amounts of the microspheres are the same as in Example 3. Table 7 Components of Comparative Examples 1-6

[0055] Comparative Example 7: The preparation method is the same as that of Example 3, but the mass ratio of each component in the formulation is as follows: 0.5 parts of barium sulfate drug, 1.5 parts of filler, 0.5 parts of modified pore-forming agent mixture, 0.5 parts of modified adhesive mixture, and 0.5 parts of modified acoustic response agent.

[0056] Comparative Example 8: The preparation method is the same as that of Example 3, but the mass ratio of each component in the formulation is as follows: 0.5 parts of barium sulfate drug, 1.5 parts of filler, 0.5 parts of modified pore-forming agent mixture, 1.5 parts of modified adhesive mixture, and 0.9 parts of modified acoustic response agent.

[0057] Experimental Example 1 In vitro drug release rate and ultrasound-responsive release experiment (1) Experimental preparation: Formulation samples: Microparticles prepared in Examples 1-6 and Comparative Examples 1-8 (using barium sulfate as the model drug); Release media: Simulated gastric juice (pH 1.2, containing 0.1M HCl), phosphate buffer (PBS, pH 6.8); Instruments: Intelligent drug release instrument, UV-Vis spectrophotometer, and ultrasonic instrument (frequency 2.5MHz).

[0058] (2) Experimental methods: 1) In vitro sustained-release experiment (without sonication) Step 1: Take 100mg of each preparation sample and place it in 500mL of pH1.2 simulated gastric juice, and shake at 37℃ (100rpm).

[0059] Step 2: Take 5 mL samples at time points of 0.5h, 1h, 2h, 4h, 6h, 8h, and 12h, and simultaneously add an equal volume of release medium at the same temperature.

[0060] Step 3: The sample was filtered through a 0.45 μm filter membrane, and the absorbance of barium sulfate in the filtrate was measured using UV-Vis. The cumulative release rate was then calculated.

[0061] 2) Ultrasonic response release experiment Step 1: Take 100 mg of each of the microparticles prepared in Examples 1-6 and Comparative Examples 1-8, place them in 500 mL of simulated gastric fluid at pH 1.2, shake at 37°C for 1 h (without ultrasound), and record the initial release rate.

[0062] Step 2: Apply 2.5MHz ultrasonic treatment to the system (power 1W / cm², time 5min), continue oscillation and take samples at 0.5h, 1h and 2h after ultrasonication to determine the cumulative release rate.

[0063] (3) Experimental data are shown in Table 8. In vitro release data for Example 1 are shown in Table 8. Figure 1 As shown, the in vitro release data for Example 2 are as follows: Figure 2 As shown, the in vitro release data for Example 3 are as follows: Figure 3 As shown, the in vitro release data for Example 4 are as follows: Figure 4 As shown, the in vitro release data for Example 5 are as follows: Figure 5 As shown, the in vitro release data for Example 6 are as follows: Figure 6 As shown, the comparative in vitro release data are as follows: Figure 7 As shown: Table 8. In vitro release data

[0064] In Example 3, the release rate increased from 17.6% to 65.7% within 2 hours after ultrasonic treatment, and the release rate was significantly accelerated, confirming that the modified acoustic response agent (calcium carbonate + coated Poria cocos extract) can trigger drug release through ultrasound. Although Comparative Example 3 (without modified acoustic response agent) showed an increase in release, the increase was smaller than that in Example 3, indicating that the modification of the acoustic response agent by the coated Poria cocos extract can enhance ultrasound sensitivity.

[0065] Experimental Example 2 In vitro gastric mucosal adhesion measurement (1) Experimental materials: Formulation samples: 100 mg of each of the microcapsules prepared in Examples 1-6 and Comparative Examples 1-8; Gastric mucosa model: fresh pig gastric mucosa (muscle layer removed, cut into 1 cm × 1 cm thin slices); Instruments: micro-force sensor (accuracy 0.1 mN), constant temperature incubator (37℃).

[0066] (2) Experimental methods: 1) Sample pretreatment: The microspheres were moistened with phosphate buffer (pH 6.8) for 10 min to simulate the gastric environment; 2) Adhesion force determination: The porcine gastric mucosa was fixed on a glass slide and placed in an incubator at 37°C; the microparticles were pressed onto the mucosa surface using a micro-force sensor (pressure 10g, contact time 30s), and the sensor was slowly and vertically pulled up to record the maximum separation force (mN). Each group of samples was measured 10 times and the average value was taken.

[0067] (3) Experimental data are shown in Table 9, and adhesion force measurement data are shown in Table 9. Figure 8 As shown: Table 9 Adhesion force measurement data

[0068] The adhesion force of Example 3 reached 12.6 mN, significantly higher than that of Comparative Example 2 (using only the modified porogen) and Comparative Example 5 (using only the modified adhesive mixture), confirming that Ca 2+ The dual network structure of ionic crosslinking and -SS-covalent bonds can synergistically enhance adhesion. In Comparative Example 7 (where the proportion of the modified adhesive mixture was insufficient), the increase in adhesion was small, indicating that the optimization of the mass ratio of each component is crucial to the adhesion performance.

[0069] Experimental Example 3 In vivo long-acting microsphere application effects (1) Experimental materials: the formulations of the examples and comparative examples of this application, the test animals and groups: healthy adult beagle dogs, weighing 10-15kg, 4 dogs per group, 14 groups in total, provided by Slack Jingda (Shanghai) Experimental Animal Co., Ltd.; (2) Experimental protocol: Fasting for 12 hours before administration was carried out to reduce the effect of food on drug absorption and gastric emptying. The formulations of the example group and the comparative group were administered orally, with normal water intake. After 6 minutes, gastric ultrasound was performed (ultrasound frequency 2.5MHz, ultrasound time 5min). (3) Evaluation of gastric retention: After ultrasound, X-ray imaging was used to record the gastric distribution of the formulation in the example group and the comparative group 0.5 h after administration. The retention time of the formulation in the example group and the comparative group was observed and recorded every 0.5 h after administration (each divided into 10 levels, the higher the level, the more uniform the gastric distribution and the longer the retention time). The results are shown in Table 10, and the content represented by the level is shown in Table 11. Table 10 Distribution and retention time of the formulation in the stomach

[0070] It should be noted that the levels of gastric distribution effect and gastric retention time in Table 10 are the average values ​​for each group.

[0071] Table 11 shows the distribution and retention time of each level in the stomach.

[0072] (4) Experimental results: As can be seen from the results in Table 10, the distribution effect and retention time of the long-acting microspheres absorbed in the stomach of the example group of this application are significantly higher than those of the comparative group, and the effects of Example 3 and Example 4 are the best.

[0073] In summary, this long-acting microsphere exhibits sustained-release characteristics in in vitro release experiments through the synergistic effect of a modified pore-forming agent mixture, a modified adhesive mixture, and a modified acoustic responsive agent. The release rate is significantly accelerated after ultrasound triggering. In vitro adhesion force measurements show that its adhesion force is significantly improved compared to traditional microspheres. In vivo experiments confirm that it is evenly distributed in the stomach and has a long retention time, which can effectively enhance the treatment effect of gastric diseases. It has the advantages of sustained-release, targeted absorption, and responsive drug release.

[0074] Although embodiments of this application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of this application. All equivalent changes and improvements made within the scope of this application shall still fall within the patent coverage of this application.

Claims

1. A long-acting pellet suitable for gastric absorption, characterized in that, The long-acting pellets are made of components in the following mass ratio: 0.01-1 parts of a drug, 1-2.5 parts of a filler, 0.2-0.7 parts of a modified pore-forming agent mixture, 1-2.5 parts of a modified adhesion agent mixture, and 0.1-0.7 parts of a modified phonophoresis agent, wherein the modified pore-forming agent mixture is made of a modified pore-forming agent and a glycosylated saponin in a mass ratio of 1:1; the modified adhesion agent mixture is formed of an adhesion agent mixture and CaCl2 by ionic cross-linking to form a covalent ionic double network; and the modified phonophoresis agent is prepared from a phonophoresis agent and coated Poria cocos extract in a mass ratio of 9:

1.

2. The long-acting pellets for gastric absorption according to claim 1, wherein the pellets are coated with a coating layer comprising a water-soluble polymer and a water-soluble organic acid. The drug is a gastritis drug, wherein the gastritis drug is any one of omeprazole, rabeprazole, clarithromycin, and amoxicillin; and the filler is any one of microcrystalline cellulose, lactose, and pregelatinized starch.

3. The long-acting pellets for gastric absorption according to claim 1, wherein the pellets are coated with a coating solution containing a water-soluble polymer and a water-soluble organic solvent. The modified pore-forming agent is a modified pore-forming agent mixed with selenoamino acid in a mass ratio of 4:1, the pore-forming agent is any one of polyethylene glycol 2000, sodium dodecyl sulfate, and polyethylene glycol 3000, and the glycosylated saponin is a product of β-glucosidase hydrolysis of tianjiang saponin. The preparation method of the modified pore-forming agent mixture is as follows: (1) Glycosylated saponin preparation: dissolve 1-5 mg / mL of substrate tianjiang saponin, 5-10 eq of p-nitrophenyl-β-D-glucoside, β-glucosidase (10-20 U / mg substrate), and tert-butyl alcohol-water (4:1 v / v) in sodium acetate buffer, and perform TLC monitoring at 50°C in the dark for 2 h, add an equal volume of methanol, perform ice bath for 10 min, centrifuge at 10,000 rpm for 10 min at 4°C, take the supernatant for concentration, add ethyl acetate for extraction to obtain crude glycosylated saponin, and perform silica gel column purification to obtain purified glycosylated saponin; (2) Modified pore-forming agent preparation: mix any one of the pore-forming agents polyethylene glycol 2000, sodium dodecyl sulfate, and polyethylene glycol 3000 with selenoamino acid in a mass ratio of 4:1, and dissolve in deionized water until completely dissolved to obtain the modified pore-forming agent; (3) Mixing: dissolve the modified pore-forming agent and the glycosylated saponin in 30% ethanol solution in a mass ratio of 1:1, and use ultrasonic-assisted dissolution to form a clear solution; the parameters are 40 kHz, 30 min, and 25°C; (4) Mixed micelle self-assembly: maintain the solution under magnetic stirring (500 rpm, 25°C) for 2 h to combine the hydrophobic parts of the modified pore-forming agent and the glycosylated saponin, and spontaneously form mixed micelles to obtain the modified pore-forming agent mixture; (5) Purification and drying: load the mixed solution into a dialysis bag, dialyze with deionized water for 24 h (every 4 h, change the water), remove the uncombined modified pore-forming agent and free glycosylated saponin, and freeze-dry (-80°C pre-freezing, 24 h freeze-drying) to obtain solid mixed micelle powder; (6) Characterization: observe the micelle morphology by electron microscopy to be spherical or near-spherical.

4. The long-acting pellets for gastric absorption according to claim 1, wherein the pellets are coated with a coating solution containing a water-soluble polymer and a water-soluble organic solvent. The adhesive mixture is prepared by mixing any one of adhesive polyacrylic acid, chitosan, carrageenan, sodium alginate, carbomer, hydroxypropyl methyl cellulose and BHP-SH with a mass ratio of 4:1, wherein the BHP-SH is obtained by grafting L-cysteine on extract of wax gourd (BHP).

5. The long-acting pellets for gastric absorption according to claim 4, wherein the enteric coating is a cellulose acetate phthalate. The preparation method of the improved adhesive mixture and the adhesive mixture is as follows: (1) Preparation of wax gourd extract: 1) Homogenization: cut and homogenize or ultra-micro-pulverize the wax gourd pulp to destroy the cell wall and release more pectin and polysaccharides; 2) Heat extraction: high-temperature cooking (80-100℃) treatment to promote the dissolution of pectin and polysaccharides and enhance the hydration; after cooling, a weak gel may be formed; 3) Freeze-thaw cycle: repeatedly freeze (-20℃) and thaw (room temperature) the wax gourd pulp to destroy the cell structure, and the ice crystal formation makes the polysaccharide network looser, and the porosity increases after water absorption; 4) Compound enzymolysis: add pectinase (0.5% w / v) + cellulase (0.2% w / v), pH 4.5, hydrolyze at 50℃ for 2h, and then inactivate the enzyme at 90℃ for 10min; after cooling, dialysis (10kDa molecular weight cut-off) is performed to purify the BHP; (2) Preparation of adhesive mixture: 1) BHP pretreatment: dissolve the purified BHP in 0.1M PBS buffer (pH=6.0) to prepare a 5% (w / v) solution, stir overnight at 4℃ to completely dissolve, and centrifuge (8000rpm, 15min) to remove insoluble substances; 2) Carboxyl activation: add EDC·HCl (2 times the molar amount of BHP carboxyl group) and NHS (1.2 times the molar amount of EDC) to step (1), stir under nitrogen protection at 25℃ for 2h in the dark, and maintain the pH at 6.0±0.2; 3) Mercapto reaction: add L-cysteine (1.5 times the molar amount of BHP carboxyl group), react under nitrogen protection at 40℃ water bath for 6h, and maintain the pH at 5.0±0.2; 4) Purification: transfer the reaction solution of step (3) to a dialysis bag, dialyze with 1mM EDTA in 0.1M NaCl solution (24h) and deionized water (48h, 6 times of water change), and freeze-dry to obtain BHP-SH white powder; 5) Dissolve any one of adhesive polyacrylic acid, chitosan, carrageenan, sodium alginate, carbomer, hydroxypropyl methyl cellulose and BHP-SH in step (4) in deionized water with a mass ratio of 4:1, and mix and dry to obtain the adhesive mixture; (3) Preparation of improved adhesive mixture: 1) Ca 2+ Network formation: Adhesive mixture was dissolved in 5 mL deionized water, 1 mL of 2% CaCl2 solution was added, and the mixture was gently stirred (200 rpm) for 30 min under nitrogen protection to avoid bubble generation. The mixture was left to stand for 1 h to allow Ca 2+ Uniform diffusion crosslinking; 2) Covalent crosslinking: The mixture was transferred to a silica gel dish and placed in a 37 °C incubator for 6 h to promote spontaneous oxidation of -SH to -S-S-; after gel formation, unreacted Ca 2+ ; 3) Drying: freeze-drying (-80℃ pre-freezing, vacuum drying for 24h) to obtain a porous sponge-like improved adhesive mixture.

6. The long-acting pellet for gastric absorption according to claim 1, wherein the long-acting pellet for gastric absorption is a pellet having a diameter of 0.1 to 2 mm. The sonodynamic response agent is any one of calcium carbonate, sodium carbonate or sodium bicarbonate, and the coating Poria cocos extract is prepared by mixing Poria cocos extract and ethyl cellulose with a mass ratio of 3:

1.

7. The long-acting pellet for gastric absorption according to claim 1, wherein the long-acting pellet for gastric absorption is a pellet having a diameter of 0.1 to 2 mm. The preparation method of the improved sonodynamic response agent is as follows: (1) Dissolve ethyl cellulose: mix ethyl cellulose and anhydrous ethanol at a ratio of 1:10 (w / v), and stir magnetically for 2h until completely dissolved to obtain an EC ethanol solution (10% w / v); (2) Coating process: Put the Poria cocos extract into the fluidized bed coating machine, preheat the material to 40℃, spray EC ethanol (100g EC dissolved in 1L ethanol), inlet air temperature 50℃, atomization pressure 0.8bar, flow rate 5mL / min, after coating, dry at 40℃ for 2h, pass through 80 mesh sieve, obtain coated Poria cocos extract; (3) Modified sonophoresis response agent: Take the sonophoresis response agent and the coated Poria cocos extract with a mass ratio of 9:1, put them into the three-dimensional mixer, mix for 30min (rotation speed 20rpm), homogenize, dry at 40℃ for 4h under vacuum, obtain the modified sonophoresis response agent.

8. A method of preparing a long-acting pellet suitable for gastric absorption, characterized by, The preparation method is a wet granule preparation method and a fluidized bed preparation method.

9. The method for preparing a long-acting microsphere suitable for gastric absorption as described in claim 8, characterized in that, The wet granule preparation method steps are: (1) Grind the drug and the filler into powder respectively, pass through 60-150 mesh sieve, mix thoroughly and uniformly; (2) Mix the drug and filler powder of step (1) with the pore-forming agent mixture, the modified adhesion agent mixture, and the modified sonophoresis response agent according to the mass ratio of claim 1; (3) Spray 5%-50% ethanol solution to make soft material; (4) Pass the soft material through a sieve with a pore size of 2-3mm to make wet granules; (5) Dry the wet granules in a drying device to remove water; (6) Pass the dried granules through a 1-2mm sieve to make whole granules, seal and package.

10. The method for preparing a long-acting microsphere suitable for gastric absorption as described in claim 8, characterized in that, The fluidized bed preparation method steps are: (1) Grind the drug and the filler into powder respectively, pass through 60-150 mesh sieve, mix thoroughly and uniformly; (2) Mix the drug and filler powder of step (1) with the pore-forming agent mixture, the modified adhesion agent mixture, and the modified sonophoresis response agent according to the mass ratio of claim 1; (3) Put into the fluidized bed, complete the granulation-drying-whole granulation process, seal and package.

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