A gastric acid resistant multi-layer core-shell microcapsule composition for targeted eradication of helicobacter pylori and its preparation dosage form and process
Patent Information
- Application Number
- CN202610912535.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-06-24
- Publication Date
- 2026-09-25
AI Technical Summary
构建五层固定顺序核壳结构微囊，耦合 GSNO 定点硅烷化 + Zn²⁺螯合、噬菌体抗酶解交联 + 保守靶点靶向修饰、竹多糖钙离子交联缓冲、PHB 阈值型 pH 响应控释、外层安全解毒多重技术特征，解决胃酸降解、酶解失效、靶向性差、休眠菌残留、亚硝胺安全隐患问题，提供可工业化量产胶囊、饮料剂型及稳定可重复制备工艺
本专利所有量化参数 ±20% 浮动区间，由 GSNO 键能特性、PHB 分子响应阈值、多糖交联网络力学性能、噬菌体蛋白耐受范围、人体胃内 pH 动态波动范围共同确定，区间内调整仅为常规工艺优化，不产生新的技术效果，适用等同侵权判定。
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Abstract
Description
Technical Field
[0001] This invention belongs to the technical fields of biological antibacterial compositions, targeted sustained-release formulations, and functional food preparation, specifically involving a five-layer acid-resistant core-shell microcapsule composition, molecular modification methods, pH adaptive controlled-release process, Helicobacter pylori targeted eradication formulation, and industrial preparation scheme. Background Technology
[0002] Helicobacter pylori is a Gram-negative pathogen that colonizes the mucus layer of the human gastric mucosa and is a high-risk factor for chronic gastritis, peptic ulcers, and gastric cancer. Clinical quadruple antibiotic therapy has drawbacks such as the increasing rate of bacterial resistance, intestinal flora imbalance, heavy metabolic burden on the liver and kidneys, and high relapse rate after drug withdrawal. Existing GSNO anti-Helicobacter pylori preparations mostly use ordinary polymer physical encapsulation without site-specific molecular structure reinforcement. Their decomposition rate exceeds 80% within 2 hours at pH 1.5, and they are rapidly oxidized and inactivated by NO. Existing phage anti-Hp preparations only perform conventional capsid cross-linking, making them easily degraded by pepsin, dependent on the LPS receptor on the outer membrane of easily mutable Hp, with a narrow serotype compatibility range, and unable to permanently inhibit urease. Direct use of natural fresh bamboo sap polysaccharides in combination results in poor resistance to mechanical shearing, easy destruction by food components, and low bamboo flavonoid activity, easily inducing single-point drug resistance in strains. Single PHB pH-responsive microcapsules lack a multi-layered, hierarchical protective structure, cannot simultaneously resist gastric acid, pepsin, and gastric peristalsis shearing, and lack the ability to eliminate dormant bacteria and block nitrosamines. Existing technologies are all single-step improvements and cannot simultaneously meet the five requirements of stable strong acid environment, targeted drug release at lesions, broad-spectrum coverage of clinical strains, eradication of dormant strains, and safety for long-term use. The various technical aspects cannot achieve a synergistic effect. Comparison of existing technologies in terms of quantitative effects: Ordinary GSNO-encapsulated formulations at pH 1.5 showed a decomposition rate of >80% in gastric juice over 2 hours; conventional phage formulations showed a capsid integrity rate of less than 40% over 3 hours in the stomach; and monolayer pH-responsive microcapsules showed an in vivo effective utilization rate of <15%. Summary of the Invention
[0003] 3.1 Purpose of the Invention We constructed a five-layered, fixed-sequence core-shell microcapsule, coupled with multiple technical features including GSNO site-directed silanization + Zn²⁺ chelation, phage anti-enzymatic crosslinking + conserved target-targeting modification, bamboo polysaccharide calcium ion crosslinking buffer, PHB threshold-type pH-responsive controlled release, and outer layer safety detoxification. This addresses issues such as gastric acid degradation, enzymatic failure, poor targeting, dormant bacteria residue, and nitrosamine safety hazards, providing industrially scalable capsule and beverage dosage forms, as well as a stable and reproducible preparation process. 3.2 Mechanism of Action GSNO γ-position silanization creates a steric barrier, preventing H⁺ from approaching the sulfur atom, while Zn²⁺ coordination reduces the electron cloud shift of the S-NO bond, thus doubly reducing the probability of strong acid breakage. Using materials with a molecular weight of 8-100,000 PHB and a film thickness of 80-120 nm achieves dense barrier at pH < 3.0 and loose, open-pore drug release at pH ≥ 5.2. Bamboo polysaccharide and sodium alginate cross-linking form a three-dimensional network to resist mechanical shearing, while internal sodium bicarbonate constructs a local micro-pH buffer environment. Esterified bamboo flavonoids enhance Hp proton pump inhibitory activity. Phage capsid cross-linking blocks pepsin hydrolysis sites, resulting in an Hp0379 protein mutation rate of <0.1%, enabling broad-spectrum targeting via scFv. Ni chelating agent permanently blocks the nickel active site of urease, preventing UreH repair protein dissociation. NO damages the biofilm EPS. The matrix utilizes bacteriophage nucleases to degrade the genome of dormant bacteria, modified phenolic acids to disrupt cell wall peptidoglycan, and a three-tiered pathway to clear dormant bacteria. Excessive molar ratios of tea polyphenol derivatives quench nitrosamines, and low-dose smooth muscle modulators locally antagonize bloating without affecting overall gastrointestinal motility. All five layers are indispensable; removal of any one layer results in a functional decrease of over 30%. The coupling of these components produces unexpected synergistic effects. 3.3 Scientific Basis for Parameter Floating Range The ±20% fluctuation range of all quantitative parameters in this patent is determined by the bond energy characteristics of GSNO, the PHB molecular response threshold, the mechanical properties of the polysaccharide cross-linking network, the tolerance range of phage proteins, and the dynamic fluctuation range of pH in the human stomach. Adjustments within this range are merely routine process optimizations and do not produce any new technical effects, thus qualifying for equivalent infringement. Attached Figure Description Figure 1 This is a schematic diagram of the radial cross-sectional structure of the five-layer core-shell microcapsule of the present invention; 1 is the modified GSNO sustained-release core, 2 is the PHB hydrophobic pH-responsive sealing layer, 3 is the calcium ion cross-linked fresh bamboo sap polysaccharide gel layer, 4 is the scFv targeted glacial phage functional layer, and 5 is the safe detoxification sustained-release outer layer. Figure 2 This is a schematic diagram of the microcapsule's adaptive and targeted drug release principle in the stomach based on pH gradient in the gastric cavity; 1 represents the strongly acidic environment region of the gastric cavity, 2 represents the weakly acidic environment region of the gastric mucosa, 3 represents the multi-layered core-shell microcapsule as a whole, 4 represents the pH-responsive sealed layer, 5 represents the drug core, and 6 represents the gastric mucosal tissue. Figure 3 This is a schematic diagram of the three-stage process by which the composition of the present invention removes dormant Helicobacter pylori encapsulated in a biofilm; 1 is the biofilm outer layer destruction process, 2 is the dormant Helicobacter pylori targeted removal process, and 3 is the biofilm regeneration inhibition and consolidation process. Figure 4This is a schematic diagram of the industrial preparation process of the multilayer core-shell microcapsule composition of the present invention; 1 is the raw material pretreatment process, 2 is the chemical reaction process, 3 is the separation and purification process, 4 is the finished product drying process, and 5 is the finished product packaging process.
Claims
1. A multilayered core-shell microcapsule composition for targeted eradication of Helicobacter pylori with gastric acid resistance, characterized in that, The composition comprises a fixed, irreversible core-shell structure with five layers from the inside out. The order of the layers cannot be changed. The layers are: a modified GSNO sustained-release core, a PHB hydrophobic pH-responsive sealed middle layer, a calcium ion cross-linked fresh bamboo sap polysaccharide gel shear-resistant layer, a scFv targeted modified glacial phage functional layer, and a safe, detoxifying, and sustained-release outer layer. The five-layer structure synergistically achieves: a stable gastric environment with no significant leakage at pH 1.2–2.5; targeted drug release at pH ≥ 5.2 in the Hp colonization area; irreversible inactivation of Helicobacter pylori urease; breaking down the ammonia-neutral microenvironment barrier of the bacteria; lysing and encapsulating dormant Helicobacter pylori in the proliferative and biofilm states; and in-situ quenching of nitrosamine ions to block the formation of nitrosamines. The five-layer structure is a necessary technical feature to achieve all the above technical effects. Removing any layer will result in a decrease of at least 30% or more in at least one core performance. Adjustments to the quantitative parameters within ±20% of this patent and replacement with equivalent materials that meet the performance indicators are all within the scope of equivalent protection.
2. The composition according to claim 1, characterized in that, The modified GSNO sustained-release core uses GSNO γ-glutamic acid side chain amino-site silanization modification combined with Zn²⁺ bidentate chelation modification, and the modification site does not touch the S-NO functional bond; The silanizing agent is methyltrimethoxysilane. The molar ratio of GSNO to the silanizing agent is 1:0.85, with a fluctuation range of 0.68–1.
02. The molar ratio of GSNO to zinc acetate chelation is 2:1, with a fluctuation range of 1.6–2.
4. After incubation in artificial gastric juice at pH 1.5 for 2 hours, the GSNO decomposition rate is ≤9.5%, with an upper limit of ≤12%, which is still within the protection range. Same-site modification with short-chain alkylsilanes and same-mode chelation of S-NO bonds with divalent metal ions are equivalent technical solutions.
3. The composition according to claim 1, characterized in that, The PHB hydrophobic pH-responsive sealed layer thickness is based on 80–120 nm, with a fluctuation range of 64–144 nm; when pH < 3.0, the hydrogen ion permeability is based on ≤3%, with an upper limit of ≤5%; when pH ≥ 5.2, it automatically forms microporous sustained-release drugs, triggering a pH of 5.2, with a fluctuation range of 4.96–5.44; polyhydroxyalkanoate materials can replace PHB and meet the above permeability and pH trigger threshold parameters, which are equivalent technical solutions.
4. The composition according to claim 1, characterized in that, The calcium ion crosslinked fresh bamboo sap polysaccharide gel layer is formed by blending low-temperature refined fresh bamboo sap polysaccharide and sodium alginate at a mass ratio of 1:0.3, and then crosslinking with calcium chloride. The ratio fluctuates between 0.8 and 1.2:0.24 and 0.
36. The gel is doped with 5% micron-sized sodium bicarbonate by mass, with a fluctuation of 4% to 6%, to create an in-situ micro-buffer environment with a pH of 3.6 to 5.
4. Bamboo flavonoids and bamboo phenolic acids are modified by acetic anhydride esterification to reduce Hp and proton pump inhibition IC50. Alginate and pectin-based natural polymers are compounded to enhance the shear resistance of the polysaccharide, which is an equivalent technical solution.
5. The composition according to claim 1, characterized in that, Preparation of scFv-targeted modified glacial phage functional layer: 0.05% glutaraldehyde was used to crosslink the β-sheet hydrolysis sites of the phage capsid at 4℃ in the dark for 45 min. The volume fraction of glutaraldehyde was 0.04%–0.06%, and the crosslinking time was 36–54 min. After crosslinking, the integrity rate of the phage after incubation with artificial gastric juice and pepsin for 3 h was ≥91% at the baseline and ≥80% at the lower limit. The phage capsid was coupled with scFv targeting Hp0379 in an EDC / NHS system, with a coupling rate of ≥85% at the baseline and ≥68% at the lower limit. Ni ion chelating urease irreversible inhibitors were simultaneously coupled. Highly conserved Hp outer membrane protein targeting peptides replaced scFv, and similar irreversible urease inhibitors replaced existing reagents. These are all equivalent technical solutions.
6. The composition according to claim 1, characterized in that, The safe, detoxifying, and sustained-release outer layer is composed of tea polyphenol derivatives and small molecules that regulate gastrointestinal smooth muscle at a mass ratio of 85:15, with a fluctuation range of 68:32 to 102:
18. The tea polyphenol derivatives quench free nitrosamine ions, and the smooth muscle regulator locally antagonizes the gastric emptying delay and bloating effect caused by NO. Natural polyphenolic antioxidants equivalently replace tea polyphenol derivatives, and similar mild gastrointestinal smooth muscle regulators replace existing small molecules, which is an equivalent technical solution.
7. An anti-Helicobacter pylori enteric-coated capsule prepared from the composition of any one of claims 1 to 6, characterized in that, Each capsule contains 450mg and uses No. 0 HPMC enteric-coated capsule shells. The formula includes: 315mg of five-layer core-shell composite microcapsule powder, 80mg of microcrystalline cellulose (pH 102), 30mg of fructooligosaccharides, 17mg of croscarmellose sodium, and 8mg of magnesium stearate. The microcapsule powder accounts for 70% of the total mass of the capsules, with a fluctuation range of 56% to 84%. Pharmaceutical-grade fillers, lubricants, disintegrants, and enteric gelatin shells are used to replace HPMC enteric-coated shells. The capsule size is scaled up proportionally, all within the protected range. The capsules do not disintegrate in gastric juice at pH 1.2 to 2.5 for 3 hours, and the Hp colonization area rapidly disintegrates and releases the microcapsules.
8. A Helicobacter pylori maintenance functional beverage prepared from the composition of any one of claims 1 to 6, wherein the finished product formula per 500 mL is: 4.2 g of four-layer simplified acid-resistant microcapsule particles, 12 g of erythritol, 0.8 g of sodium citrate, 0.35 g of tragacanth gum, 0.15 g of vitamin C palmitate, and purified water to a final volume; the mass fluctuation of each component is ±20%, and the replacement excipients of similar food-grade stabilizers and sweeteners are all protected; the finished product has a pH of 3.7 to 3.9 and is aseptically cold-filled; any oral dosage form prepared using five-layer core-shell microcapsules as the core active ingredient, such as granules, suspensions, lozenges, chewable tablets, or oral emulsions, is included within the scope of protection of this patent.
9. The preparation process of the multilayer core-shell microcapsule composition according to any one of claims 1 to 6, comprising the following steps in sequence: 1) completing the site-directed silanization of GSNO and Zn²⁺ chelation reaction under nitrogen atmosphere at 0-4℃, and dialysis purification to prepare modified core microparticles; 2) homogenizing molten PHB with core microparticles, and spraying the PHB layer by fluidized bed atomization, controlling the film thickness to 80-120nm; 3) preparing calcium ion crosslinked fresh bamboo sap polysaccharide gel at low temperature, doping with sodium bicarbonate microparticles, and secondary coating in a fluidized bed; 4) completing phage capillary crosslinking, scFv coupling, and urease irreversible inhibitor linkage at low temperature; 5) spraying a composite outer layer of tea polyphenol derivative and smooth muscle regulator in a fluidized bed; 6) freeze drying, sieving microcapsules with a particle size of 200-250μm, and vacuum sealing in the dark; the above steps are necessary characteristics of the process, and the operation time ±20%, slight temperature fluctuations, and replacement of conventional industrial equipment do not change the essence of the process and fall within the scope of process protection. This process involves temperature and humidity control throughout. Except for the PHB melting section, the ambient temperature of the remaining processes is ≤8℃, with an upper limit of fluctuation of ≤9.6℃; the relative humidity of the production environment is ≤40%, with an upper limit of fluctuation of ≤48%, and the entire process is carried out in a light-proof and oxygen-free environment.
10. The application and patented equipment protection scope of the five-layer core-shell microcapsule composition according to claims 1 to 6 are as follows: This composition can be used in five scenarios: routine Helicobacter pylori eradication, intervention of multi-drug resistant Helicobacter pylori, clearance of dormant Helicobacter pylori biofilm strains, auxiliary repair of gastric mucosa in Hp-related chronic gastritis, and prevention of recurrence after Helicobacter pylori seroconversion. The application protection premise is that the product has the fixed five-layer core-shell structure and corresponding molecular modification features of this patent. At the same time, it is specially adapted to the customized fluidized bed, low-temperature reactor, and high-pressure homogenization equipment modified with the 80-120nm spraying parameters of the PHB layer, the GSNO fixed-point modification low-temperature reaction station, and the phage scFv low-temperature coupling chamber. Any use of this patented product for large-scale production constitutes indirect infringement.