Probiotic gastric floating sustained-release tablet capable of dispelling effects of alcohol and preparation method of probiotic gastric floating sustained-release tablet

Abstract

The invention relates to an anti-alcoholism probiotic gastric floating sustained-release tablet and a preparation method thereof in the technical field of pharmaceutical preparation preparation. The anti-alcoholism probiotic gastric floating sustained-release tablet comprises a mucous membrane attaching layer and an acid-sensitive release layer, the mucosa attaching layer is used for realizing targeted adhesion and retention of the tablet on the surface of gastric mucosa and providing a gastric acid blocking function; the acid-sensitive release layer is used for quickly responding and releasing probiotics in a gastric acid environment so as to realize local efficient alcoholism relieving in the stomach. Through the multi-layer structure of the floating sustained-release tablet, the tablet floats after being used before meals and is firmly fixed on the surface of gastric mucosa through the mucosa attaching layer, and the release holes ensure that probiotics are quickly released in the stomach, so that the probiotics are quickly released in a gastric acid environment and locally act in the stomach, the activity of the probiotics is protected, and the effect of preventing and treating the gastric mucosa is achieved. Meanwhile, the sustained release effect is achieved by means of retention of the tablet in gastric mucosa, so that the local efficient, rapid and continuous hangover alleviating effect in the stomach is achieved, and interference to intestinal flora is reduced.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, specifically a probiotic gastric floating sustained-release tablet for relieving hangovers and its preparation method. Background Technology

[0002] After alcohol is consumed, it is absorbed by the stomach but primarily metabolized in the liver through an enzymatic system, converted into acetaldehyde, then into acetic acid, and finally broken down into carbon dioxide and water and excreted from the body. Many hangover remedies contain traditional Chinese medicine ingredients, which may have effects such as strengthening the spleen and removing dampness, aiding digestion and resolving phlegm, clearing heat and detoxifying, refreshing the mind, and neutralizing alcohol. Some hangover products contain peptides or proteins, which may help promote alcohol metabolism. For hangover remedies to function, ingredients that help accelerate alcohol breakdown, alleviate post-drinking discomfort, or protect the body from alcohol damage are required.

[0003] Existing technologies include the preparation of hangover remedies using freeze-dried powders formulated with different types of probiotics, as shown in patent number CN115944087A. These primarily utilize strains that efficiently express alcohol dehydrogenase and aldehyde dehydrogenase, maximizing the protection of the freeze-dried probiotics' activity through freeze-drying protectants. This allows the probiotics to convert unabsorbed ethanol into acetaldehyde, which in turn is converted into acetic acid, thus achieving a hangover-relieving and sobering effect. However, these freeze-dried probiotic powder preparations rely on intestinal colonization, leading to a delayed onset of action. This not only carries the risk of disrupting the intestinal flora balance but also fails to rapidly exert its effects in the peak alcohol absorption areas of the stomach and upper small intestine.

[0004] Therefore, it is necessary to propose a gastric floating sustained-release probiotic tablet for hangover relief and its preparation method that can overcome the problems of slow onset of action, insufficient protection of bacterial activity, and inability of immediate-release formulations to provide continuous protection. Summary of the Invention

[0005] To address the aforementioned problems, the present invention aims to provide a gastric floating sustained-release tablet for relieving hangovers and its preparation method. Through the multi-layered structure of the floating sustained-release tablet, the mucosal adhesion layer allows the tablet to float and firmly adhere to the gastric mucosa surface after pre-meal use. The release pores ensure rapid release of probiotics within the stomach, allowing the probiotics to be rapidly released in the acidic gastric environment and act locally within the stomach, protecting the activity of the probiotics. Simultaneously, the sustained-release effect is achieved through the tablet's retention on the gastric mucosa, thereby achieving a highly efficient, rapid, and sustained hangover-relieving effect locally within the stomach, while reducing interference with the intestinal flora.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: a probiotic gastric floating sustained-release tablet for relieving hangovers, comprising a mucosal adhesion layer and an acid-sensitive release layer; The mucosal adhesion layer is used to achieve targeted adhesion and retention of tablets on the gastric mucosa surface and to provide gastric acid barrier function; The acid-sensitive release layer is designed to respond quickly to and release probiotics in the acidic environment of the stomach, thereby achieving highly efficient local detoxification of alcohol in the stomach.

[0007] The probiotic gastric floating sustained-release tablets for hangover relief achieve targeted and efficient hangover relief in the stomach through the synergistic effect of the mucosal adhesion layer and the acid-sensitive release layer: The mucosal adhesion layer uses adhesive materials to target and adhere the tablet to the gastric mucosa surface, prolonging the retention time in the stomach and blocking gastric acid, thus protecting the internal components; The acid-sensitive release layer responds quickly in the acidic environment of the stomach and releases active probiotics, ensuring that the probiotics are released in a concentrated manner in the stomach; After release, the probiotics form a high concentration distribution on the gastric mucosa surface, accelerating alcohol metabolism by producing acetaldehyde dehydrogenase and inhibiting abnormal metabolism of harmful bacteria to reduce the stimulation of toxic products. Ultimately, it achieves the effect of local degradation of alcohol in the stomach and reduction of damage before alcohol absorption, solving the pain points of probiotics being easily killed by gastric acid, easily lost with gastric emptying, and difficult to take effect locally.

[0008] Furthermore, the mucosal adhesion layer comprises the following materials in parts by weight: 6-15 parts of carbomer 974P, 15-16 parts of poloxamer 407, 43-45 parts of hydrophobically modified light microcrystalline cellulose, 3-4 parts of sodium bicarbonate, 5-6 parts of calcium carbonate, 6-8 parts of crospovidone, and 2.5-3.5 parts of sodium stearate fumarate.

[0009] Carbomer 974P and poloxamer 407 enhance adhesion. Combined with the lightweight properties of hydrophobically modified lightweight microcrystalline cellulose and the gas buoyancy of sodium bicarbonate and calcium carbonate reacting with gastric acid, they counteract gastric emptying, prolonging the tablet's residence time in the stomach and laying the foundation for its action. Sodium bicarbonate and calcium carbonate neutralize local gastric acid, working in conjunction with the physical barrier of hydrophobically modified lightweight microcrystalline cellulose to provide dual protection for the acid-sensitive release layer and probiotics, reducing the risk of their inactivation by gastric acid. Cross-linked povidone improves tablet structural stability and disintegration efficiency, while sodium stearate fumarate improves material flowability and compressibility, enhancing formulation quality, production feasibility, and product stability.

[0010] Furthermore, the probiotic freeze-dried powder in the acid-sensitive release layer is encapsulated by polyvinyl acetate phthalate to form acid-sensitive microcapsules. The acid-sensitive release layer includes the following materials in parts by weight: 28-32 parts probiotic freeze-dried powder, 19-23 parts mannitol, 8-11 parts trehalose, 30-45 parts hydroxypropyl methylcellulose K4M, and 8-10 parts polyvinyl acetate phthalate.

[0011] The lyophilized probiotic powder is encapsulated in acid-sensitive microcapsules using polyvinyl acetate phthalate. Combined with the protective effects of mannitol and trehalose, this process resists the influence of preparation, storage, and the acidic environment of the stomach, reducing the probiotic inactivation rate and ensuring its activity. Polyvinyl acetate phthalate rapidly dissolves in stomach acid, releasing the probiotics. Hydroxypropyl methylcellulose (K4M) regulates the release rate, achieving a stable and continuous release within the stomach, forming a localized high-concentration distribution to act on alcohol. K4M enhances the material's binding properties, improving tablet forming quality. Mannitol improves the material's flowability and compressibility, reducing production problems and ensuring the stability of the formulation's physical properties.

[0012] Furthermore, the probiotic freeze-dried powder contains the following probiotics in parts by weight: Lactobacillus fermentum CBT LF1 45-50 parts, Lactobacillus acidophilus CBT LA1 20-30 parts, Bifidobacterium bifidum CBT BB1 15-25 parts and Saccharomyces boulardii CBT SB1 15-15 parts.

[0013] While existing probiotic preparations for relieving hangovers can degrade alcohol by expressing ADH and ALDH enzymes, they still have significant drawbacks. In addition to relieving hangovers, it is necessary to improve the survival rate of probiotics, reduce their impact on the balance of intestinal flora, and alleviate the burden on the stomach, intestines, and liver.

[0014] The *Lactobacillus* strain selected in this invention is CBT LF1, which has excellent alcohol dehydrogenase (ADH) activity, enabling it to efficiently convert ethanol into acetaldehyde; *Lactobacillus acidophilus* strain is CBT LA1, which has extremely strong gastric acid tolerance, significantly improving the survival rate of the entire bacterial community in the stomach; *Bifidobacterium bifidum* strain is CBT BB1, which has high acetaldehyde dehydrogenase (ALDH) activity, responsible for rapidly converting acetaldehyde (the main source of toxicity) into non-toxic acetic acid; and *Saccharomyces boulardii* strain is CBT SB1, which can secrete multiple enzymes (including ADH and ALDH) to assist in the degradation of alcohol, significantly enhancing intestinal barrier function, reducing intestinal leakage and endotoxin entry into the bloodstream caused by alcohol, alleviating systemic inflammatory response and liver burden, effectively neutralizing bacterial toxins, and improving gastrointestinal discomfort symptoms that may occur after drinking alcohol.

[0015] The above four probiotics are combined in a specific ratio. Lactobacillus fermentum CBT LF1 is the core producer of metabolic enzymes, while other strains assist in enhancing enzyme activity, optimizing degradation pathways, and broadening the scope of action, synergistically improving the rate of alcohol degradation in the stomach. Lactobacillus fermentum and Lactobacillus acidophilus form a physical barrier, Bifidobacterium bifidum regulates inflammation, and Saccharomyces boulardii adsorbs toxins and promotes repair, protecting the gastric mucosa on multiple levels and reducing the risk of alcohol-induced damage. The probiotic combination competitively inhibits the growth of harmful bacteria, regulates the bacterial community structure, and inhibits toxin release, restoring the balance of the gastric microecology and reducing secondary damage from alcohol metabolism. The strains form an acid-resistant alliance, working in conjunction with the mucosal adhesion layer and acid-sensitive release layer to improve the retention rate of activity during storage and gastric transport, ensuring a high live bacteria count to exert the effects of detoxification and stomach protection.

[0016] Furthermore, the acid-sensitive release layer is wrapped in the mucosal adhesion layer, and several release holes are opened in the center of the mucosal adhesion layer on both the upper and lower surfaces. The release holes correspond to the acid-sensitive release layer, and the outer wall of the mucosal adhesion layer has wavy patterns that match the folds of the gastric mucosa.

[0017] This invention fixes the mucosal adhesion layer and the acid-sensitive release layer together, resisting the impact of gastric peristalsis, avoiding separation failure, and ensuring a stable combination of retention barrier and precise release. The release pores in the center of the mucosal adhesion layers on both the upper and lower surfaces provide a directional channel for probiotics, ensuring that they quickly and concentratedly reach the gastric mucosal surface and form a high concentration distribution. The wavy texture adapts to the folds of the gastric mucosa, increasing the contact area and fitting into gaps, strengthening adhesion, and prolonging the residence time in the stomach. The texture fits together to form a closed space, strengthening the gastric acid barrier. The release pores are directly aligned with the mucosa, reducing the loss of probiotics and enhancing the local targeting of alcohol detoxification and gastric protection.

[0018] Furthermore, the hardness of the sustained-release tablet is 75-85 N.

[0019] This hardness range balances the physical stability and functionality of tablets: ensuring they are not easily broken during production and transportation, maintaining structural integrity to protect the internal probiotics; while avoiding excessive hardness that hinders normal disintegration and release in the gastric environment, ensuring the adhesion performance of the mucosal adhesion layer and the release efficiency of probiotics in the acid-sensitive release layer, while also meeting the needs of industrial production, improving product qualification rate and reliability.

[0020] This application also provides a method for preparing the above-mentioned probiotic gastric floating sustained-release tablets for relieving hangovers, including the following steps: Step 1: Preparation of acid-sensitive release layer particles: a. Mix the bacterial sludge of Lactobacillus fermentum CBT LF1, Lactobacillus acidophilus CBT LA1, Bifidobacterium bifidum CBT BB1 and Saccharomyces boulardii CBT SB1 in a certain proportion, mix it evenly with mannitol and trehalose, and freeze-dry to obtain compound probiotic freeze-dried powder; b. Using the emulsification-solvent evaporation method, the composite probiotic freeze-dried powder obtained in step a is encapsulated in polyvinyl acetate phthalate to prepare acid-sensitive microcapsules; c. Mix the acid-sensitive microcapsules obtained in step b with hydroxypropyl methylcellulose K4M evenly, add the first binder solution to make a soft material, granulate, dry, and granulate to obtain inner layer particles. Step 2: Preparation of mucosal adhesion layer particles: Carbomer 974P, hydrophobically modified lightweight microcrystalline cellulose, sodium bicarbonate, calcium carbonate, and cross-linked polyvinyl ketone were mixed evenly, and a second binder solution was added to form a soft material. The material was then granulated, dried, and sized. Sodium stearate fumarate was added and mixed evenly to obtain the outer layer particles. Step 3: Tableting: Using a tablet press, the outer layer particles are first filled into the mold cavity and lightly pressed to form the base and release hole. Then, the inner layer particles are filled onto the mucosal adhesion layer base and lightly pressed to shape. Finally, the outer layer particles are added again and pressed into shape under a pressure of 8-10kN to obtain the sobering probiotic gastric floating sustained-release tablet.

[0021] Furthermore, the specific parameters of the emulsification-solvent evaporation method described in step 1b are as follows: polyvinyl acetate phthalate is dissolved in anhydrous ethanol as the oil phase, and 0.5% polyvinyl alcohol aqueous solution is used as the aqueous phase. The mixture is ultrasonically emulsified at 300W power for 5 minutes, the solvent is evaporated at 30℃, and the acid-sensitive microcapsules are collected by centrifugation.

[0022] Furthermore, in step 1c, the first adhesive is an ethanol solution of povidone K30, and its amount is 4-6% of the total weight of the inner layer dry particles.

[0023] Furthermore, in step two, the second adhesive is an ethanol solution of hydroxypropyl methylcellulose E50, and its amount is 2-4% of the total weight of the outer dry particles.

[0024] Furthermore, the hardness of the sustained-release tablet is 75-85 N.

[0025] The above methods can produce acid-sensitive microcapsules with uniform particle size and high encapsulation rate, which can not only avoid mechanical damage to probiotics during subsequent granulation and tableting, but also achieve precise release in the gastric acid environment, solving the problem of easy inactivation of live bacteria in existing formulations. Layered filling and gradient tableting not only ensure that the mucosal adhesion layer and the acid-sensitive release layer are symmetrically fixed and structurally stable, but also make the release pores precisely correspond to the acid-sensitive release layer, avoiding the outer layer from hindering the release of probiotics. From the ultrasonic power and temperature of the emulsification-solvent evaporation method, to the type and amount of adhesive, and to the tableting pressure and filling sequence, there are specific standards, which can reduce the impact of human operation differences on product quality. Granulation and tableting are carried out using conventional formulation equipment, without the need for special customized equipment, which lowers the production threshold and is conducive to achieving large-scale mass production, providing process support for the actual promotion and application of this hangover relief sustained-release tablet.

[0026] The technical solution of the present invention has at least the following advantages and beneficial effects: (1) This invention uses gastric floating sustained-release technology to allow the tablets to remain in the stomach and continuously release active probiotics, thereby achieving rapid and direct metabolism of alcohol during the peak period of alcohol absorption, fundamentally overcoming the disadvantage of slow onset of action.

[0027] (2) This invention significantly improves the survival rate and colonization ability of live bacteria under gastric acid through a dual protection strategy of complex microbial community + intelligent gel barrier (HPMC / poloxam / carbomer), ensuring that a sufficient amount of live bacteria reach the site of action.

[0028] (3) This invention, through the synergistic combination of multiple strains, not only accelerates the complete metabolism of alcohol, but also takes into account liver protection and gastrointestinal health, providing a comprehensive and fundamental solution for detoxification and body protection.

[0029] (4) The present invention provides a continuous drug release for more than 12 hours through an optimized gastric floating sustained-release system, achieving long-term protection with a single dose, and meeting the needs of actual drinking scenarios.

[0030] (5) This invention improves the stability of the formulation and the feasibility of room temperature storage and transportation by optimizing the freeze-drying process and powder direct pressing technology, and removing moisture and temperature-sensitive excipients from the formulation, which is more conducive to commercial application. Attached Figure Description

[0031] Figure 1 This is a growth curve of compound probiotics in ethanol at different concentrations.

[0032] Figure 2 This is a standard curve of ethanol concentration.

[0033] Figure 3 The curves show the degradation rate of compound probiotics in ethanol at different concentrations over time.

[0034] Figure 4 A bar chart showing the recovery time of the righting reflex in intoxicated rats.

[0035] Figure 5 Axonometric view of a probiotic gastric floating sustained-release tablet for relieving hangovers.

[0036] Figure 6 A top view of a probiotic gastric floating sustained-release tablet for relieving hangovers.

[0037] Reference numerals: 1. Mucosal adhesion layer; 2. Release pore; 3. Acid-sensitive release layer. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the present invention will be further described below in conjunction with specific embodiments. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents used, or any products for which the manufacturer is not specified, are all commercially available conventional products. All features disclosed in this specification, except for mutually exclusive features and / or steps, may be combined in any manner.

[0039] The following embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way.

[0040] The specific implementation method is described below with reference to the accompanying drawings.

[0041] Example 1 This embodiment provides a probiotic gastric floating sustained-release tablet for relieving hangovers, such as... Figure 5 and Figure 6 As shown, it includes a mucosal adhesion layer 1 and an acid-sensitive release layer 3 wrapped around the mucosal adhesion layer 1; a plurality of release holes 2 are provided on the upper and lower surfaces of the mucosal adhesion layer 1 at positions corresponding to the acid-sensitive release layer 3.

[0042] The mucosal adhesion layer 1 is used to achieve targeted adhesion and retention of tablets on the gastric mucosa surface and to provide gastric acid barrier function. The mucosal adhesion layer 1 comprises the following materials in parts by weight: 6 parts of carbomer 974P, 15 parts of poloxamer 407, 45 parts of hydrophobically modified light microcrystalline cellulose, 3 parts of sodium bicarbonate, 6 parts of calcium carbonate, 7 parts of crospovidone, and 3.5 parts of sodium stearate fumarate.

[0043] The acid-sensitive release layer 3 is used to rapidly respond to and release probiotics in the gastric acid environment to achieve localized and efficient alcohol detoxification in the stomach. The probiotic freeze-dried powder in the acid-sensitive release layer 3 is encapsulated by polyvinyl acetate phthalate to form acid-sensitive microcapsules. The acid-sensitive release layer 3 includes the following materials in parts by weight: 30 parts probiotic freeze-dried powder, 21 parts mannitol, 10 parts trehalose, 42 parts hydroxypropyl methylcellulose K4M and 8 parts polyvinyl acetate phthalate.

[0044] The probiotic freeze-dried powder contains the following probiotics in parts by weight: Lactobacillus fermentum CBT LF1 48 parts, Lactobacillus acidophilus CBT LA1 20 parts, Bifidobacterium bifidum CBT BB1 17 parts and Saccharomyces boulardii CBT SB1 15 parts.

[0045] This embodiment also provides a method for preparing the above-mentioned probiotic gastric floating sustained-release tablets for relieving hangovers, including the following steps: Step 1: Preparation of acid-sensitive release layer 3 particles: a. Mix the bacterial sludge of Lactobacillus fermentum CBT LF1, Lactobacillus acidophilus CBT LA1, Bifidobacterium bifidum CBT BB1 and Saccharomyces boulardii CBT SB1 in a certain proportion, mix it evenly with mannitol and trehalose, and freeze-dry to obtain compound probiotic freeze-dried powder; The culture conditions for the above-mentioned probiotics are as follows: (1) Lactobacillus fermentum CBT LF1 culture: culture medium: MRS medium; culture temperature: 37±1℃; culture method: anaerobic; culture time: 18-24 hours (until the early stage of stable growth); collection method: centrifugation, 6000×g, 10 minutes, 4℃. (2) Lactobacillus acidophilus CBT LA1 culture: culture medium: MRS medium; culture temperature: 37±1℃; culture method: anaerobic; culture time: 20-24 hours; collection method: centrifugation, 6000×g, 15 minutes, 4℃. (3) Bifidobacterium bifidum CBT BB1 culture: culture medium: modified MRS medium (0.05% cysteine ​​hydrochloride added to MRS base); culture temperature: 37±1℃; culture method: anaerobic; culture time: 24-48 hours; collection method: centrifugation, 5000×g, 15 minutes, 4℃. (4) Culture of Saccharomyces blazei CBT SB1: Culture medium: YPD medium; Culture temperature: 30±1℃; Culture method: aerobic (speed 150-200 rpm); Culture time: 36-48 hours (until the growth is stable); Collection method: centrifugation, 4000×g, 5 minutes, 4℃.

[0046] After cultivation, the supernatant was poured into a waste container in a sterile operating table, and the precipitated bacterial sludge was resuspended with physiological saline. An appropriate amount of 10% mannitol was added as a freeze-drying protectant, and the mixture was transferred to a freeze dryer at -80 ℃ for 6 hours. The powder in the sterile centrifuge tubes is the probiotic freeze-dried powder.

[0047] b. Using the emulsification-solvent evaporation method, the lyophilized compound probiotic powder obtained in step a was encapsulated in polyvinyl acetate phthalate to prepare acid-sensitive microcapsules. Polyvinyl acetate phthalate was dissolved in anhydrous ethanol as the oil phase and 0.5% polyvinyl alcohol aqueous solution was used as the aqueous phase. The mixture was ultrasonically emulsified at 300W for 5 min, the solvent was evaporated at 30℃, and the acid-sensitive microcapsules were collected by centrifugation. c. Mix the acid-sensitive microcapsules obtained in step b with hydroxypropyl methylcellulose K4M evenly, add the first binder solution to form a soft material, granulate, dry, and granulate to obtain inner layer particles. The first binder is an ethanol solution of povidone K30, and its amount is 4-6% of the total weight of the dry inner layer particles. Step 2: Preparation of mucosal adhesion layer 1 particles: Carbomer 974P, hydrophobically modified lightweight microcrystalline cellulose, sodium bicarbonate, calcium carbonate, and cross-linked polyvinyl ketone were mixed evenly. A second binder solution was added to form a soft mass, which was then granulated, dried, and sized. Sodium stearate fumarate was added and mixed evenly to obtain outer layer particles. The second binder was an ethanol solution of hydroxypropyl methylcellulose E50, and its amount was 2-4% of the total weight of the dry outer layer particles. Step 3: Tableting: Using a tablet press, the outer layer particles are first filled into the mold cavity and lightly pressed to form the base and release hole 2. Then, the inner layer particles are filled onto the base of the mucosal adhesion layer 1 and lightly pressed to set the shape. Finally, the outer layer particles are added again and pressed into shape under a pressure of 8-10kN to obtain the sobering probiotic gastric floating sustained-release tablet.

[0048] Examples 2-9 Examples 2-9 each provide a probiotic gastric floating sustained-release tablet for relieving hangovers and its preparation method. The only difference from Example 1 is that the dosages of HPMC K4M, LM-MCC, poloxamer, and carbomer 974P are different from those in Example 1. The dosages and preparation methods of the remaining components are the same as in Example 1.

[0049] The dosages of HPMC K4M, LM-MCC, poloxamer, and carbomer 974P in Examples 2-9 are shown in Table 1: Table 1. Distribution ratio of each group in Examples 2-9

[0050] Experimental Example 1: Probiotics' tolerance to ethanol Experimental methods: 1. Probiotic culture (1) Culture of Lactobacillus fermentum CBT LF1: Culture medium: MRS medium; Culture temperature: 37±1℃; Culture method: anaerobic; Culture time: 18-24 hours (until the early stage of stable growth); Collection method: centrifugation, 6000×g, 10 minutes, 4℃.

[0051] (2) Culture of Lactobacillus acidophilus CBT LA1: Culture medium: MRS medium; Culture temperature: 37±1℃; Culture method: anaerobic; Culture time: 20-24 hours; Collection method: centrifugation, 6000×g, 15 minutes, 4℃.

[0052] (3) Culture of Bifidobacterium bifidum CBT BB1: Culture medium: modified MRS medium (0.05% cysteine ​​hydrochloride added to MRS base); Culture temperature: 37±1℃; Culture method: anaerobic; Culture time: 24-48 hours; Collection method: centrifugation, 5000×g, 15 minutes, 4℃.

[0053] (4) Culture of Saccharomyces blazei CBT SB1: Culture medium: YPD medium; Culture temperature: 30±1℃; Culture method: aerobic (speed 150-200 rpm); Culture time: 36-48 hours (until the growth is stable); Collection method: centrifugation, 4000×g, 5 minutes, 4℃.

[0054] The probiotics were cultured under the above conditions until their OD600 reached approximately 0.1. The bacterial concentration in the culture medium was standardized, and a compound probiotic strain was prepared according to the specified ratio (Lactobacillus fermentum CBT LF1 (45-50%), Lactobacillus acidophilus CBT LA1 (20-30%), Bifidobacterium bifidum CBT BB1 (15-25%), and Saccharomyces boulardii CBT SB1 (5-15%)). Different doses of ethanol or acetaldehyde were added to the culture medium to achieve ethanol concentrations of 0%, 2%, 4%, 6%, 8%, and 10%, respectively. The OD600 values ​​of each culture medium were recorded every 2 hours to compare the growth of the compound probiotic strains under different ethanol concentrations.

[0055] Experimental results: The results are as follows Figure 1 As shown, by plotting the growth curves of the compound probiotics in culture media containing different concentrations of ethanol, it can be seen that although there is some inhibition at 4%–6%, it can still enter the exponential growth phase; even at high concentrations of 8%–10%, there is no complete inhibition, showing a certain degree of tolerance; demonstrating the tolerance of the compound probiotic strains to ethanol.

[0056] Experiment Example 2: Detection Experiment of the In Vitro Alcohol Detoxification Ability of Compound Probiotics Experimental methods: 1. Establishment of ethanol concentration standard curve (potassium dichromate-concentrated sulfuric acid method) (1) Preparation of 0.25 mM potassium dichromate solution: Accurately weigh 7.35 mg of potassium dichromate (K2Cr2O7) and dissolve it in 80 mL of ultrapure water, then make up to 100 mL to obtain a 0.25 mM potassium dichromate solution. Take 50 mL of this solution and slowly add 50 mL of concentrated sulfuric acid (98%) under ice bath conditions while stirring to prepare a potassium dichromate-concentrated sulfuric acid working solution (prepare immediately before use).

[0057] (2) Preparation of ethanol standard solutions: Prepare 0%, 5%, 10%, 15%, 20%, 25%, and 30% (v / v) ethanol gradient solutions using anhydrous ethanol and ultrapure water. The specific preparation method is as follows: 0% group (pure water), 5% group (5 mL ethanol + 95 mL water), 10% group (10 mL ethanol + 90 mL water), and so on to prepare each concentration gradient.

[0058] (3) Construction of standard curve reaction system: Take 1 mL centrifuge tube, add 900 μL of potassium dichromate-concentrated sulfuric acid working solution to each tube, and then add 100 μL of ethanol standard solution of different concentrations (0-30%). Mix well immediately and react in the dark for 10 minutes.

[0059] (4) Absorbance determination: Using a UV spectrophotometer, the absorbance value (OD600) of each reaction solution was measured at a wavelength of 600 nm with the 0% ethanol group (blank control) zeroed. Three replicates were set for each concentration group, and the average value was taken.

[0060] (5) Standard curve plotting: Using the final ethanol concentrations (0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%) as the ordinate and the corresponding OD600nm values ​​as the abscissa, a standard curve equation (y=ax+b) was established using linear regression analysis. The correlation coefficient R² ≥ 0.99 was required. The results are as follows: Figure 2 As shown.

[0061] 2. Evaluation of the in vitro hangover-relieving effect of probiotics (1) Preparation of experimental materials: Logarithmic growth phase probiotics (OD600=0.6-1.0) were used. 0%, 2%, 4%, 6%, 8%, and 10% ethanol gradient solutions were prepared according to the instructions. Simulated gastric juice with pH=2.0 (containing 0.3% pepsin) was prepared. Potassium dichromate working solution (0.25mM K2Cr2O7: concentrated sulfuric acid=1:1) was prepared. A 37℃ constant temperature shaker (100rpm), centrifuge, and spectrophotometer were prepared.

[0062] (2) Construction of incubation system: Add 3 mL of simulated gastric fluid and 1 mL of probiotic culture (final concentration approximately 10) to each 5 mL sterile centrifuge tube. 8 The experiment used 1 mL of ethanol solution with different concentrations (CFU / mL) to achieve final ethanol concentrations of 0%, 1%, 2%, and 4%, with three replicates for each group.

[0063] (3) Setting incubation conditions: Place the mixture in a constant temperature shaker at 37℃ and shake at 100rpm to simulate gastric peristalsis. Take samples at 0, 0.5, 1, 2 and 4 hours respectively. Immediately after sampling, stop the reaction with an ice bath.

[0064] (4) Sample processing procedure: Take 500 μL of the mixture at each time point, centrifuge at 10,000 rpm for 2 min at 4℃, take the supernatant and dilute it 5 times with pre-cooled PBS, store at -20℃ for testing (not exceeding 24 hours), and restore to room temperature before testing.

[0065] (5) Detection of ethanol concentration: Take 100 μL of diluted sample and add 900 μL of potassium dichromate working solution. After vortexing and mixing, react in the dark for 10 min and immediately measure the OD600 value. Calculate the ethanol concentration of each sample according to the pre-established standard curve.

[0066] (6) Data analysis method: Calculate the ethanol residual rate and degradation rate at each time point, plot the time-ethanol concentration change curve, and compare the degradation efficiency of different concentration groups.

[0067] Experimental results: The results are as follows Figure 3 As shown, by plotting the degradation rate of ethanol by the compound probiotics in different concentrations of ethanol over time, the degradation rate was 68-70% in 1-2% ethanol and 59% in 4% ethanol. The probiotics showed degradation ability in the 1-4% ethanol concentration range.

[0068] Based on the in vitro alcohol detoxification ability test of compound probiotics in Example 1, a mixed bacterial solution with a different proportion of probiotics than that in Example 1 was prepared, and a comparative experiment on the in vitro alcohol detoxification ability test of compound probiotics was conducted, as shown in Table 2 below: Table 2. Design of bacterial suspension ratios with different characteristics

[0069] The experimental results are shown in Table 3 below: Table 3. Comparison of alcohol degradation efficiency

[0070] Experimental results showed that the bacterial strain ratio groups in Example 1 exhibited the highest degradation rates at 2% and 4% ethanol concentrations, reaching 70.2% and 59.0%, respectively, significantly outperforming other ratio combinations. In the high-proportion single-strain group, although the excessively high proportion of Lactobacillus fermentum CBT LF1 possessed strong ADH activity, the lack of synergistic effects from other strains led to a decrease in overall acid resistance and acetaldehyde conversion capacity, resulting in a significant reduction in degradation rate.

[0071] In the group with an imbalanced bacterial strain ratio, the excessive proportion of Lactobacillus acidophilus, while improving the survival rate in the acidic gastric environment, resulted in insufficient enzymatic conversion of ethanol and acetaldehyde, limiting degradation efficiency. Although the group without Saccharomyces boulardii still exhibited some ability in ethanol degradation, the lack of Saccharomyces boulardii's coenzyme secretion and intestinal barrier protection function meant that its overall effect on alcohol detoxification and gastric mucosal protection was not as good as the control group in Example 1.

[0072] Experiment Example 3: Study on the effect of probiotic gastric floating sustained-release tablets for relieving hangovers on intoxicated rats Experimental methods: SD rats (220±20g) were randomly divided into 4 groups (n=8): blank group (administered by gavage with physiological saline), alcohol model group (administered by gavage with ethanol + blank floating tablet), free probiotic group (administered by gavage with ethanol + free probiotics (equal bacterial count)), and gastric floating sustained-release tablet group (administered by gavage with ethanol + the hangover-relieving probiotic sustained-release floating tablet (50mg / kg) from Example 1). An alcohol intoxication model was established 30 minutes after administration. The alcohol intoxication model was established by gavage with 5g / kg of 40% ethanol solution (diluted with physiological saline). The time required for the righting reflex to disappear and recover was observed and recorded. The time for the righting reflex to disappear was recorded as the intoxication time, and the time required for the righting reflex to recover was recorded as the sobering-up time. Analysis of variance was used to compare the administered group with the control group to obtain the results.

[0073] Experimental results: The results are as follows Figure 4 As shown, the effect of probiotic gastric floating sustained-release tablets on relieving alcohol poisoning was evaluated by establishing an acute alcohol poisoning model in SD rats. The experimental results showed that the righting reflex recovery time in the gastric floating sustained-release tablet group (68.3±5.5 min) was significantly shorter than that in the free probiotic group (98.3±6.5 min, *p*<0.01) and the alcohol model group (135.0±7.1 min, *p*<0.001), demonstrating that this formulation has a superior effect on relieving alcohol poisoning. The gastric floating sustained-release technology significantly improves the survival rate and duration of action of probiotics by prolonging the gastric retention time and protecting them from gastric acid damage, thereby accelerating alcohol metabolism. Compared with free probiotics, the probiotic gastric floating sustained-release tablets exhibit more stable sustained-release characteristics and higher bioavailability, verifying the feasibility and advantages of this formulation in the application of alcohol relieving.

[0074] The specific implementation process of the gastric floating sustained-release tablet is as follows: After the user takes the gastric floating sustained-release tablet of the present invention, within 15 minutes, the tablet enters the stomach along with the stomach contents. At pH 1.2-3.5, the sodium bicarbonate and calcium carbonate in the mucosal adhesion layer 1 rapidly react with the gastric acid. NaHCO3+ HCl→NaCl + CO3↑ + H2O; CaCO3+ 2HCl→CaCl3+ CO3↑ + H2O; The generated CO2 forms tiny bubbles inside the tablet; at the same time, the low density of the hydrophobically modified lightweight microcrystalline cellulose, with a bulk density of <0.3g / cm³, further reduces the overall density of the tablet, allowing it to float quickly on the surface of the stomach contents and preventing it from entering the intestines with the stomach emptying.

[0075] Carbomer 974P in the mucosal adhesion layer 1 swells upon contact with gastric acid, forming an adhesive gel layer that binds to mucin on the surface of the gastric mucosa via hydrogen bonds. Simultaneously, the wavy texture on the outer wall of the tablet adapts to the folded structure of the gastric mucosa, thereby increasing the contact area and firmly fixing the tablet to the surface of the gastric mucosa, precisely covering the core area of ​​alcohol absorption. Furthermore, the dense structure of the mucosal adhesion layer 1 can physically block gastric acid from directly eroding the inner acid-sensitive release layer 3, providing the first protective barrier for probiotics.

[0076] Over the next 12 hours, the polyvinyl acetate phthalate in the acid-sensitive release layer 3 is a gastric acid-sensitive material that dissolves rapidly in the acidic environment of the stomach, exposing the encapsulated lyophilized probiotic powder. At the same time, the hexagonal 3mm diameter release hole 2 in the center of the mucosal adhesion layer 1 is precisely aligned with the acid-sensitive release layer 3, ensuring that the dissolved probiotics can directly act on the stomach through the release hole 2, avoiding obstruction by the outer particles and achieving rapid release of bacteria into the stomach.

[0077] The hydroxypropyl methylcellulose K4M in the acid-sensitive release layer 3 forms a viscous gel skeleton when it encounters gastric acid, which slowly dissolves and releases probiotics. By regulating the gel dissolution rate, the compound probiotics are continuously released within 12 hours, with a cumulative release rate of 72%. This process can match the alcohol absorption cycle of long-term drinking, avoiding the problem of diminishing effect after a single release of bacteria, and achieving continuous hangover protection.

[0078] The released probiotics maintain high activity in the stomach through the freeze-drying protectant and the acid-resistant properties of Lactobacillus acidophilus. Lactobacillus fermentum CBT LF1 efficiently secretes alcohol dehydrogenase (ADH), which catalyzes the conversion of unabsorbed ethanol in the stomach into acetaldehyde. C2H5OH + NAD + → CH3CHO + NADH + H + ; Bifidobacterium bifidum CBT BB1 secretes aldehyde dehydrogenase (ALDH), which further converts the more toxic acetaldehyde into non-toxic acetic acid. CH3CHO + NAD + + H2O → CH3COOH + NADH + H + , Acetic acid is eventually metabolized by the liver into CO2 and H2O and excreted from the body.

[0079] The acid-resistant properties of Lactobacillus acidophilus CBT LA1 can improve the survival rate of the entire bacterial community in the stomach, ensuring that the enzymatic reaction continues. Saccharomyces boulardii CBT SB1 secretes additional ADH and ALDH to help break down alcohol, while also enhancing intestinal barrier function, reducing alcohol-induced intestinal mucosal leakage, lowering the risk of endotoxins entering the bloodstream, reducing the metabolic burden on the liver, and relieving discomfort such as fatigue and bloating after drinking.

[0080] The compound probiotics are released and exert their effects directly in the peak area of ​​alcohol absorption in the stomach, avoiding the delayed onset of action of existing intestinal colonization preparations, and can significantly shorten the sobering time after drinking. The probiotics only act locally in the stomach and do not need to colonize the intestines, thus avoiding intestinal flora imbalance from the root. The preparation does not contain analgesics, diuretics, or other ingredients, and does not burden the liver and kidneys for metabolism. The freeze-drying process and acid-sensitive microencapsulation allow the probiotics to retain more than 85% of their activity during storage at room temperature and away from light for 6 months, eliminating the need for cold chain transportation and making it convenient for daily use.

[0081] Experiment Example 4: Performance Indicators of Probiotic Gastric Floating Tablets for Hangover Relief The following indicators of the probiotic gastric floating tablets prepared in Examples 1-9 were determined: floating lag time (min), total floating time (h), cumulative release rate over 12 hours (%), and tablet hardness (N).

[0082] Floating performance test: Place the tablet in 200mL of artificial gastric fluid at 37℃ and record the time required for complete floating and the continuous floating time. The qualified standard is: lag time ≤5min and total floating time ≥12h.

[0083] Release rate determination: A dissolution apparatus (USPII method, paddle plate) was used with 900 mL of artificial gastric fluid (pH 1.2) as the medium, at 50 rpm and 37 ± 0.5 °C. Samples of 5 mL were taken at 0.5, 1, 2, 4, 6, 8, and 12 h (HPLC analysis of probiotic release), and the solution was replenished. A cumulative release rate of 60-80% after 12 h was required.

[0084] Hardness test: The radial crushing force of the tablets (n=10) is tested using a hardness tester. The acceptable range is 70-90N, ensuring that the tablets are not easily damaged during transportation and storage.

[0085] The experimental results are shown in Table 4. Table 4. Performance Indicators of Probiotic Gastric Floating Tablets for Hangover Relief

[0086] In summary, the probiotics for relieving hangovers prepared by this invention all have good buoyancy and ideal sustained-release characteristics, while also possessing suitable mechanical strength, meeting the quality requirements of gastric floating sustained-release tablets.

[0087] This invention is not limited to the above-described embodiments. Anyone should know that any structural changes made under the guidance of this invention, and any technical solutions that are the same as or similar to this invention, fall within the protection scope of this invention.

Claims

1. A probiotic gastric floating sustained-release tablet for relieving hangovers, characterized in that, It includes a mucosal adhesion layer (1) and an acid-sensitive release layer (3); The mucosal adhesion layer (1) is used to achieve targeted adhesion and retention of tablets on the gastric mucosa and to provide gastric acid barrier function; The acid-sensitive release layer (3) is used to quickly respond to and release probiotics in the gastric acid environment to achieve localized and efficient alcohol detoxification in the stomach.

2. The probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 1, characterized in that, The mucosal adhesion layer (1) comprises the following components in parts by weight: 6-15 parts of carbomer 974P, 15-16 parts of poloxamer 407, 43-45 parts of hydrophobically modified light microcrystalline cellulose, 3-4 parts of sodium bicarbonate, 5-6 parts of calcium carbonate, 6-8 parts of crospovidone, and 2.5-3.5 parts of sodium stearate fumarate.

3. The probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 1, characterized in that, The probiotic freeze-dried powder in the acid-sensitive release layer (3) is encapsulated by polyvinyl acetate phthalate to form acid-sensitive microcapsules. The acid-sensitive release layer (3) includes the following components in parts by weight: 28-32 parts of probiotic freeze-dried powder, 19-23 parts of mannitol, 8-11 parts of trehalose, 30-45 parts of hydroxypropyl methylcellulose K4M and 8-10 parts of polyvinyl acetate phthalate.

4. The probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 3, characterized in that, The probiotic freeze-dried powder contains the following probiotics in parts by weight: Lactobacillus fermentum CBT LF1 45-50 parts, Lactobacillus acidophilus CBT LA1 20-30 parts, Bifidobacterium bifidum CBT BB1 15-25 parts and Saccharomyces boulardii CBT SB1 5-15 parts.

5. The probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 2, characterized in that, The acid-sensitive release layer (3) is wrapped inside the mucosal attachment layer (1); several release holes (2) are opened in the center of the mucosal attachment layer (1) on both the upper and lower surfaces, and the release holes (2) correspond to the acid-sensitive release layer (3). The outer wall of the mucosal attachment layer (1) has a wavy pattern that matches the folds of the gastric mucosa.

6. The method for preparing the probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 5, characterized in that, The hardness of the sustained-release tablet is 75-85 N.

7. A method for preparing a gastric floating sustained-release probiotic tablet for relieving hangovers as described in any one of claims 1-6, characterized in that, Includes the following steps: Step 1: Preparation of acid-sensitive release layer (3) particles: a. Mix the bacterial sludge of Lactobacillus fermentum CBT LF1, Lactobacillus acidophilus CBT LA1, Bifidobacterium bifidum CBT BB1 and Saccharomyces boulardii CBT SB1 in a certain proportion, mix it evenly with mannitol and trehalose, and freeze-dry to obtain compound probiotic freeze-dried powder; b. Using the emulsification-solvent evaporation method, the composite probiotic freeze-dried powder obtained in step a is encapsulated in polyvinyl acetate phthalate to prepare acid-sensitive microcapsules; c. Mix the acid-sensitive microcapsules obtained in step b with hydroxypropyl methylcellulose K4M evenly, add the first binder solution to make a soft material, granulate, dry, and granulate to obtain inner layer particles. Step 2, Preparation of mucosal adhesion layer (1) particles: Carbomer 974P, hydrophobically modified lightweight microcrystalline cellulose, sodium bicarbonate, calcium carbonate, and cross-linked polyvinyl ketone were mixed evenly, and a second binder solution was added to form a soft material. The material was then granulated, dried, and sized. Sodium stearate fumarate was added and mixed evenly to obtain the outer layer particles. Step 3: Tableting: Using a tablet press, the outer layer particles are first filled into the mold hole and lightly pressed to form the base and release hole (2). Then, the inner layer particles are filled onto the base of the mucosal adhesion layer (1) and lightly pressed to shape. Finally, the outer layer particles are added again and pressed into shape under a pressure of 8-10kN to obtain the sobering probiotic gastric floating sustained-release tablet.

8. The method for preparing the probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 7, characterized in that, The specific parameters of the emulsification-solvent evaporation method described in step 1b are as follows: polyvinyl acetate phthalate is dissolved in anhydrous ethanol as the oil phase, and 0.5% polyvinyl alcohol aqueous solution is used as the aqueous phase. The mixture is ultrasonically emulsified at 300W power for 5 minutes, the solvent is evaporated at 30℃, and the acid-sensitive microcapsules are collected by centrifugation.

9. The method for preparing the probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 7, characterized in that, In step 1c, the first adhesive is an ethanol solution of polyvinyl ketone K30, and its amount is 4-6% of the total weight of the inner layer dry particles.

10. The method for preparing the probiotic gastric floating sustained-release tablet for relieving hangovers according to claim 7, characterized in that, In step two, the second adhesive is an ethanol solution of hydroxypropyl methylcellulose E50, and its amount is 2-4% of the total weight of the outer dry particles.

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