Long-acting hydrogen-releasing microspheres in the stomach and their use as medicaments in the treatment of gastritis

Abstract

The application discloses a long-acting hydrogen-releasing microsphere in the stomach and use of the microsphere as a drug in treatment of gastritis, and belongs to the field of pharmaceutical preparations and hydrogen treatment. The composition provided by the application is composed of a hydrogen production micro-particle as a core and a sustained-release agent composed of hydroxypropyl methyl cellulose and ethyl cellulose. By selecting specific components and processes, the microsphere has good floatability and long-acting hydrogen-releasing capacity. The microsphere can float in the gastric juice environment and gradually release hydrogen, so that the sustained anti-oxidation and anti-inflammatory effects are realized, and the microsphere is suitable for treatment of gastric inflammation.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparations and hydrogen therapy, specifically relating to a method for preparing long-acting hydrogen-releasing microspheres that float in gastric juice and their application in the treatment of gastritis. Background Technology

[0002] Gastritis is a common inflammation of the gastric mucosa, usually caused by Helicobacter pylori infection, overuse of medications (such as NSAIDs), irregular eating habits, and stress. These pathogenic factors lead to oxidative stress and inflammatory responses, thereby damaging the gastric mucosa, causing excessive gastric acid secretion and impaired mucosal barrier function. Traditional drug treatments have certain side effects and drug resistance problems. Therefore, the development of low-toxicity anti-inflammatory drugs holds promise for achieving highly effective treatment of gastritis. Hydrogen, as a safe and non-toxic therapeutic gas molecule, can play an important role in inflammatory-related diseases.

[0003] Currently, the main ways to ingest hydrogen include inhaling hydrogen, taking hydrogen-rich water orally, and injecting hydrogen-containing saline. However, inhaling hydrogen makes it difficult to accumulate hydrogen at a therapeutic concentration in the lesion site in the stomach, and taking hydrogen-rich water orally limits the dosage due to the low solubility of hydrogen molecules. Therefore, the development of a method that can slowly release hydrogen in the stomach and stay there for a long time can solve the problem of efficient treatment of gastric inflammation.

[0004] Therefore, designing a formulation that can release hydrogen gas over a long period of time and relieve gastritis symptoms has important clinical value. Summary of the Invention

[0005] This invention relates to a method for preparing long-lasting hydrogen-releasing microspheres that can float in the stomach and their application in gastritis, belonging to the pharmaceutical field. Addressing the problem that existing technologies cannot achieve long-lasting hydrogen release in the stomach, the composition provided by this invention uses hydrolyzed hydrogen-producing microparticles as the core and is coated with a sustained-release agent composed of hydroxypropyl methylcellulose and ethyl cellulose. Due to the different solubility properties of hydroxypropyl methylcellulose and ethyl cellulose in gastric juice, hydroxypropyl methylcellulose can be rapidly dissolved by gastric acid to form porous microspheres. With the infiltration of gastric acid, hydrogen is released and escaped, enabling the microspheres to float in the stomach. This prolonged retention time in the stomach extends the action time of hydrogen molecules, resulting in good biocompatibility and a high hydrogen loading capacity.

[0006] One of the objectives of this invention is to provide a method for preparing long-lasting hydrogen-releasing microspheres that can float in the stomach and their application in gastritis. The hydrogen-releasing microspheres are composed of hydrolyzable hydrogen-producing micro / nano materials and an encapsulation layer. The mass percentage of the hydrolyzable hydrogen-producing micro / nano materials is 25-75 wt%, preferably 45 wt%, and the mass percentage of the encapsulation layer is 25-75 wt%. The size of the micro / nano materials is less than 40 micrometers.

[0007] The aforementioned hydrolysis-based hydrogen production micro / nano materials include, but are not limited to, materials that can react with water to produce hydrogen, such as magnesium hydride, calcium hydride, silicon hydride, magnesium silicide, calcium silicide, cerium silicide, titanium silicide, magnesium, zinc, iron, and silicon.

[0008] The coating layer is prepared by mixing a water-insoluble sustained-release matrix material and a water-soluble sustained-release matrix material. The preferred mass ratio of the water-insoluble sustained-release matrix material to the water-soluble sustained-release matrix material is 60-100:1-40, and more preferably 80:20.

[0009] The water-insoluble sustained-release matrix materials include, but are not limited to, ethyl cellulose, polyethylene, polymethyl methacrylate Eudragit RS and Eudragit RL; the water-soluble sustained-release matrix materials include, but are not limited to, hydroxypropyl methylcellulose and polyethylene glycol.

[0010] A second objective of this invention is to provide a method for preparing long-lasting hydrogen-releasing microspheres that can float in the stomach, the method comprising the following steps: (1) Dissolve the water-insoluble and water-soluble sustained-release framework materials in non-polar and polar organic solvents, respectively, and then blend them. The solvents are water-insoluble (EC), water-soluble (HPMC), non-polar organic solvent (dichloromethane), and polar organic solvent (ethanol). The water-insoluble materials are dissolved in the non-polar organic solvent, and the water-soluble materials are dissolved in the polar organic solvent. The mass ratio requirement is 0-1:0-1:2000-4000:2000-4000.

[0011] The polar solvents include, but are not limited to, methanol, ethanol, and acetone, with ethanol being preferred; the non-polar organic solvents include, but are not limited to, chloroform, dichloromethane, benzene, and toluene, with dichloromethane being preferred.

[0012] (2) Next, add the hydrolysis hydrogen-producing micro / nano materials and mix well; The content of the hydrolysis hydrogen-producing micro / nano materials is 25-75 wt%, preferably 45 wt%.

[0013] (3) Using liquid paraffin as the oil phase, add emulsifier and mix, while stirring, slowly add the mixed solution from step (2); The emulsifier may be a Span emulsifier, a Tween emulsifier, or a poloxamer, preferably Span 83. The emulsifier accounts for 1% to 5% of the mass of the liquid paraffin, and the stirring speed is 300 to 800 rpm.

[0014] Liquid paraffin, as an oil phase, is not easily volatile and is immiscible with the aforementioned polar and non-polar organic solvents. The mixed solution in step (2) is an aqueous phase. The emulsifier has an lipophilic group on one end and a hydrophilic group on the other, which allows the mixed solution in step (2) to maintain a spherical shape in the liquid paraffin.

[0015] (4) In a well-ventilated environment, continue stirring until the solvent has completely evaporated; (5) Centrifuge to obtain solid microspheres, wash thoroughly to remove liquid paraffin and excess emulsifier, and finally vacuum dry to obtain the final product.

[0016] The third objective of this invention is the use of the aforementioned long-acting hydrogen-releasing microspheres in the stomach as a drug in the treatment of gastritis. The hydrogen-releasing microspheres are tiny particles similar to capsules, which can be made into capsules, granules, pills, or other suitable shapes.

[0017] The hydrogen-releasing microspheres (such as MgH2@EC / HPMC microcapsules) provided by this invention float for more than 6 hours in a simulated gastric fluid environment.

[0018] The hydrogen-releasing microspheres (such as MgH2@EC / HPMC microcapsules) provided by this invention can continuously release hydrogen gas for about 24 hours in a simulated gastric juice environment.

[0019] The hydrogen-releasing microspheres (such as MgH2@EC / HPMC microcapsules) provided by this invention can effectively improve gastric inflammation and repair gastric mucosal damage.

[0020] The water-insoluble sustained-release matrix material (ethyl cellulose EC) is insoluble at any pH in vivo, but it swells in the presence of gastric juice. The water-soluble sustained-release matrix material (hydroxypropyl methylcellulose HPMC) is unaffected by pH in water. When microspheres are added to simulated gastric juice, HPMC slowly dissolves, forming pores in the microspheres. Simultaneously, the gastric juice permeates into the microspheres and interacts with the hydrolytic hydrogen-decomposing material MgH within the microspheres. ２ The reaction generates hydrogen gas, which is slowly released within the swollen microspheres. Since the overall density of (hydrogen + microspheres) is less than the density of simulated gastric juice, the microspheres can float. In addition, a water-insoluble slow-release framework material retains the hydrogen; it is almost undissolved, only swollen. The presence of hydrogen reduces the overall density of the microspheres, allowing them to float. This increases the residence time of the microspheres in the stomach, prolonging the interaction time of hydrogen molecules, resulting in good biocompatibility and a high hydrogen loading capacity. The penetration of gastric acid and the hydrolysis of the hydrogen-producing micro / nanomaterials cause hydrogen molecules to escape, allowing the hydrogen-releasing microspheres to float in the stomach. The prolonged residence time in the stomach improves the bioavailability of hydrogen molecules.

[0021] Therefore, the present invention can also control the hydrogen release time and release rate by changing the amount of water-soluble sustained-release matrix material (hydroxypropyl methylcellulose HPMC). The hydrogen release time and release rate can be controlled by changing the amount of water-soluble sustained-release matrix material, and the suitable mass concentration of water-soluble sustained-release matrix material (hydroxypropyl methylcellulose HPMC) is 1~40%.

[0022] The hydrogen-releasing microspheres for treating gastritis prepared in this invention have the following advantages: 1) It exhibits good biocompatibility both in vitro and at the cellular level; 2) It has a good spherical shape, uniform size and high hydrogen loading capacity; 3) It continuously and slowly releases hydrogen gas in the acidic environment of the stomach, which prolongs the time that the drug works in the stomach. It can float and remain in the stomach for a long time. It can release hydrogen molecules in simulated gastric juice for up to 24 hours. Therefore, it can be used for long-term treatment of gastritis. Its antioxidant effect protects the gastric mucosa and relieves gastritis symptoms through its anti-inflammatory effect.

[0023] 4) Compared with methods such as inhaling hydrogen gas, drinking hydrogen water, and injecting hydrogen-rich saline, this invention can achieve a long-term sustained release of a large amount of hydrogen gas in the stomach through oral administration, thereby increasing the hydrogen dose in the stomach and the treatment time. This invention provides a method for preparing floating, long-lasting hydrogen-releasing microspheres. By selecting specific components and processes, the microspheres possess excellent buoyancy and long-lasting hydrogen release capability. The microspheres of this invention can float in the gastric juice environment and gradually release hydrogen gas, thereby achieving sustained antioxidant and anti-inflammatory effects, making them suitable for the treatment of gastric inflammation. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the structure of the microsphere composition of the present invention; Figure 2 This is a scanning electron microscope image of the microspheres of this invention; Figure 3 These are digital electronic images and particle size distribution diagrams of the microspheres of this invention; Figure 4 This is the XRD pattern of the microspheres of this invention; Figure 5 This is the infrared spectrum of the microspheres of this invention; Figure 6 This is a graph showing the hydrogen release curves of different samples; Figure 7 This is a GC chromatogram (hydrogen loading test) of the hydrogen production of the microspheres of this invention: 5 mg of floating long-acting hydrogen-releasing microspheres were added to 5 ml of artificial gastric fluid and sealed in a 20 ml vial. The result was that 5 mg of microspheres released 0.02 μmol of hydrogen gas, with a hydrogen release amount of 0.004 μmol / mg; Figure 8 This is a diagram showing the buoyancy properties of the microspheres of this invention in a simulated in vivo environment using artificial gastric fluid; Figure 9 This is a schematic diagram of cell viability and death staining after co-culturing gastric mucosal epithelial cells (GES-1) with different drug concentrations of the microspheres of the present invention for 24 hours; Figure 10 This is a graph showing the anti-inflammatory effect of different concentrations of the microspheres of the present invention on gastric mucosal epithelial cells under lipopolysaccharide (LPS)-induced inflammation. Detailed Implementation

[0025] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0026] Example 1 The floating hydrogen-releasing microspheres provided by this invention are as follows: Figure 1 As shown, the preparation steps of the floating hydrogen-releasing microspheres are as follows: (1) First, dissolve 100 mg of ethyl cellulose (EC) in 15 mL of dichloromethane and 20 mg of hydroxypropyl methylcellulose (HPMC) in 15 mL of ethanol, and then mix them together.

[0027] (2) Next, add 100 mg of magnesium hydride powder and mix well; (3) Add 1.5wt% of Span-83 to 120mL of liquid paraffin and slowly add the mixed solution from step (2) while stirring.

[0028] (4) Stir continuously for 24 hours in a well-ventilated environment until the solvent has completely evaporated; (5) The solidified MgH2@EC / HPMC microspheres were collected by centrifugation, washed three times with n-hexane, and dried under vacuum to obtain the final product.

[0029] Using the same synthesis method, microspheres with HPMC contents of 0% (as a control), 15%, and 20% were obtained.

[0030] 1. Particle size distribution of floating, long-lasting hydrogen-releasing microspheres The floating, long-lasting hydrogen-releasing microspheres (HPMC mass percentage 20%) prepared in Example 1 were photographed using an electron microscope. The microsphere diameter distribution was statistically analyzed using ImageJ software, and plotted using Origin software. Figure 3 As shown in the digital electronic diagram (left) and particle size distribution diagram (right), the floating long-lasting hydrogen-releasing microspheres prepared by this invention have a good spherical morphology, uniform size, and good dispersibility.

[0031] Figure 2 This is a schematic diagram of a floating long-lasting hydrogen-releasing microsphere provided by the present invention using a scanning electron microscope (SEM). It can be seen that its morphology is microsphere-shaped.

[0032] Figure 4 The image shows the XRD pattern of the microspheres of this invention. As can be seen from the image, the MgH2 particles generated by hydrolysis in the microspheres retain their original crystal structure, indicating that no significant structural changes occurred during the preparation process.

[0033] Figure 5 This is the infrared spectrum of the microspheres of the present invention. It can be seen from the figure that the absorption peaks of the prepared microspheres are consistent with those of the element, indicating that no impurities were introduced during the preparation process.

[0034] 2. Hydrogen production rate of floating long-lasting hydrogen-releasing microspheres 5 mg of the floating, long-acting hydrogen-releasing microspheres prepared in Example 1 were added to 5 mL of artificial gastric fluid and sealed. The hydrogen release curve of the microspheres was measured using a hydrogen electrode. Figure 6 It can be observed that from 0 to 5 hours, the hydrogen production rate of microspheres with 20% HPMC is faster, after which the hydrogen production rate gradually decreases and is released completely in a short time; while microspheres with 0% HPMC only enter a slightly faster hydrogen production rate after 12 hours, and microspheres with 15% HPMC enter a slightly faster hydrogen production rate in about 1 hour, with a long hydrogen release duration. It can be seen that by changing the amount of HPMC added, the hydrogen release rate and hydrogen release time of the microspheres can be changed accordingly.

[0035] 3. Continuous hydrogen production capacity of floating, long-lasting hydrogen-releasing microspheres Add 5 mg of the floating long-acting hydrogen-releasing microspheres prepared in Example 1 to 5 mL of artificial gastric fluid, seal the bottle, and periodically use a sampling needle to collect 50 μL of gas from the bottle. Measure the gas chromatography to obtain the GC chromatogram of the floating long-acting hydrogen-releasing microspheres prepared in Example 1. Figure 7 The result showed that 5 mg of microspheres released 0.02 micromoles of hydrogen gas, with a hydrogen release amount of 0.004 micromoles / mg. It can be seen that the amount of 20% HPMC microspheres in the bottle increases rapidly and reaches a high value in about 3 hours.

[0036] 4. Flotation performance of long-lasting hydrogen-releasing microspheres Adding 5 mg of the floating long-acting hydrogen-releasing microspheres prepared in Example 1 to 5 mL of simulated gastric fluid, and simulating intragastric buoyancy in a shaker at 37 degrees Celsius and 80 rpm, the floating long-acting hydrogen-releasing microspheres floated immediately upon being added to the simulated gastric fluid, and the floating time was greater than 5 hours. Figure 8 ).

[0037] 5. Cell-level biosafety experiments Figure 9 This diagram illustrates cell viability and mortality staining of gastric mucosal epithelial cells co-cultured at different drug concentrations (100, 200, 400, and 800 μg / ml) of the microspheres provided in this invention for 24 hours. It can be observed that the microspheres do not harm the gastric mucosal epithelial cells, and the floating, long-lasting hydrogen-releasing microspheres exhibit good biocompatibility.

[0038] 6. Anti-inflammatory effects Figure 10This image shows the anti-inflammatory effects of different concentrations of the microspheres provided by this invention on gastric mucosal epithelial cells under lipopolysaccharide-induced inflammation. Cellular inflammation was induced by co-culturing 10 ng / mL LPS and GES-1 cells for 24 h. Subsequently, inflammatory cells were co-cultured with different concentrations (100, 200, 400, and 800 μg / mL) of floating, long-acting hydrogen-releasing microspheres for 24 h. The level of cellular inflammation was detected using a ROS staining kit. It can be seen that the floating, long-acting hydrogen-releasing microspheres provided by this invention are effective in reducing inflammation.

Claims

1. A gastric long-acting hydrogen-releasing microsphere, characterized in that, The hydrogen-releasing microspheres comprise micro / nanomaterials for hydrogen production via hydrolysis and an encapsulation layer, wherein the mass percentage of the micro / nanomaterials for hydrogen production via hydrolysis is 25–75 wt%, and the mass percentage of the encapsulation layer is 25–75 wt%.

2. The gastric long-acting hydrogen-releasing microspheres as described in claim 1, characterized in that, The micro / nano materials include one or more of magnesium hydride, calcium hydride, silicon hydride, magnesium silicide, calcium silicide, cerium silicide, titanium silicide, magnesium, zinc, iron, or silicon.

3. The gastric long-acting hydrogen-releasing microspheres as described in claim 1, characterized in that, The coating layer comprises a water-insoluble sustained-release matrix material and a water-soluble sustained-release matrix material, wherein the mass ratio of the water-insoluble sustained-release matrix material to the water-soluble sustained-release matrix material is 60-100:1-40.

4. The gastric long-acting hydrogen-releasing microspheres as described in claim 3, characterized in that, The water-insoluble sustained-release matrix material is one or more of ethyl cellulose, polyethylene, polymethyl methacrylate Eudragit RS, or Eudragit RL.

5. The gastric long-acting hydrogen-releasing microspheres as described in claim 3, characterized in that, The water-soluble sustained-release matrix material is hydroxypropyl methylcellulose and / or polyethylene glycol.

6. The method for preparing gastric long-acting hydrogen-releasing microspheres according to any one of claims 1-5, characterized in that, The preparation method includes the following steps: (1) Dissolve the water-insoluble sustained-release matrix material and the water-soluble sustained-release matrix material in non-polar and polar organic solvents respectively, and then blend them together; (2) Next, add the hydrolysis hydrogen-producing micro / nano materials and mix well; (3) Using liquid paraffin as the oil phase, add emulsifier and mix, while stirring, slowly add the mixed solution from step (2); (4) In a well-ventilated environment, continue stirring until the solvent has completely evaporated; (5) Centrifuge to obtain solid microspheres, wash thoroughly to remove liquid paraffin and excess emulsifier, and finally vacuum dry to obtain the final product.

7. The method for preparing gastric long-acting hydrogen-releasing microspheres as described in claim 6, characterized in that, In step (1), the polar solvent is one or more of methanol, ethanol, or acetone, and the non-polar organic solvent is one or more of chloroform, dichloromethane, benzene, or toluene; or The emulsifier in step (3) is one or more of the following: Span emulsifier, Tween emulsifier, or poloxamer.

8. The method for preparing gastric long-acting hydrogen-releasing microspheres as described in claim 6, characterized in that, The water-soluble sustained-release matrix material in step (1) accounts for 1-40% of the total mass concentration of all sustained-release matrix materials.

9. The use of the gastric long-acting hydrogen-releasing microspheres as described in any one of claims 1-6 as a drug in the treatment of gastritis, characterized in that, The drug is an oral preparation, in the form of capsules, granules, or pills.

10. The use of the gastric long-acting hydrogen-releasing microspheres as described in claim 9 as a drug in the treatment of gastritis, characterized in that, The medication is used to treat gastritis.

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