Novel dual-phase gel microbead with hangover alleviating function and preparation method of novel dual-phase gel microbead

Abstract

The invention discloses a novel difunctional peptide fragment with antioxidant activity and alcohol metabolic enzyme activation activity. The invention also discloses a compound gel microbead containing the novel peptide fragment and having a hangover alleviating function and a preparation method of the compound gel microbead. The preparation method comprises the following steps: mixing curcumin and 2-hydroxypropyl-beta-cyclodextrin to prepare an inclusion compound; dissolving the clathrate compound and a novel peptide fragment in water to obtain a water phase; puerarin is dissolved in corn oil to obtain an oil phase, the oil phase and the water phase are mixed and homogenized at a high speed to obtain a primary emulsion, the primary emulsion and a sodium alginate solution are mixed and homogenized to form a uniform double emulsification system, and a calcium chloride solution is dropwise added to obtain the gel microbeads loaded with multiple active components. A double emulsification system is constructed by a specific method, the defect of poor water solubility of curcumin and puerarin is overcome, the novel peptide fragment is utilized to develop a novel hangover-alleviating and liver-protecting product, the three active ingredients have synergistic interaction, and meanwhile, the gel beads provide protection for the active ingredients and reduce degradation of gastric juice, so that the anti-alcoholism and liver-protecting effects are achieved, and the anti-alcoholism and liver-protecting effects are achieved. The alcoholic liver injury is effectively relieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of functional food, health products or drug delivery technology, in particular to a gel microsphere embedding multiple active ingredients and a preparation method thereof, which has the functions of alcohol elimination and liver protection. BACKGROUND

[0002] In China, the culture of drinking has existed since ancient times, but with the development of the times, the problem of diseases caused by excessive drinking has also come along, and cases of alcohol poisoning during holidays are not uncommon. Excessive intake of alcohol can stimulate the body to produce a stress response, leading to liver cell necrosis, lesions, etc. Alcoholic liver damage (ALD) refers to liver damage caused by long-term consumption of alcoholic beverages, and is the main cause of the increased incidence and mortality of liver-related diseases. The current clinical treatment methods for alcoholic liver damage mainly include alcohol withdrawal and drug therapy, but it is difficult to achieve alcohol abstinence, and long-term drug use has side effects on the body. Therefore, alcohol elimination and liver protection products with good alcohol elimination effect and no side effects have received widespread attention from the pharmaceutical and food industries.

[0003] Bioactive peptides extracted from food proteins have received increasing attention due to their various pharmacological activities. Bioactive peptides are a class of peptides that can not only scavenge oxidative free radicals in the human body, but also activate liver enzyme detoxification, and have small molecular weight, easy absorption, and health care effects of alcohol elimination. Studies have shown that peptides with biological activity extracted from food have strong antioxidant activity and can effectively alleviate alcoholic liver damage. Among the natural active ingredients for alcohol elimination and liver protection, curcumin and puerarin have been widely studied. Curcumin is the main active ingredient of turmeric, and its core function is its good antioxidant activity, which can effectively scavenge excess free radicals produced during alcohol metabolism, thereby directly reducing the oxidative damage of alcohol to liver cells; Puerarin, as the key extract of pueraria, mainly plays a role by regulating key enzymes in alcohol metabolism, which can significantly increase the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, accelerate the decomposition of ethanol into acetaldehyde, and further convert it into non-toxic acetic acid, thereby reducing the concentration of ethanol and its toxic intermediate acetaldehyde in the blood, alleviating the drunk state from the source and reducing the metabolic burden on the liver. However, the physicochemical properties of both limit their direct application effect.

[0004] In the field of oral delivery, the application of active ingredients for hangover relief and liver protection faces significant biopharmaceutical challenges. First, the acidic environment of the stomach and the various enzyme systems in the digestive tract readily cause the degradation or isomerization of phenolic substances such as curcumin and puerarin, while bioactive peptides are more easily inactivated by protease hydrolysis, leading to a significant reduction in effective dosage. Second, these ingredients often exhibit poor solubility and permeability; for example, curcumin is extremely difficult to dissolve in water, and puerarin is absorbed slowly and irregularly orally, severely limiting its bioavailability. Therefore, developing novel composite gel microbeads capable of integrating, protecting, and precisely delivering multiple functional ingredients with different physicochemical properties is key to achieving highly effective hangover relief and liver protection products.

[0005] Compared with existing technologies, this invention achieves significant innovation in functional positioning and delivery system design. For example, the currently disclosed patent "A method for preparing a liver-protecting drug composition containing dihydromyricetin" combines multiple components, and its technical focus is on the component combination itself, rather than a targeted delivery and protection system; the authorized patent "A curcumin colon-targeted drug formulation and its preparation method" achieves colon-targeted release of curcumin through advanced formulation technology, and its technical scope is limited to curcumin as a single active ingredient, with the targeting purpose being the treatment of colonic diseases; the authorized patent "A hangover relief composition and its preparation method" discloses a composition with hangover relief effect and a hangover relief beverage. In the prior art, there are no reports of combining and encapsulating curcumin, puerarin, and novel corn peptides through the "encapsulation-double emulsification-gelation" process, nor are there reports of a method for preparing composite gel microbeads with good alcohol metabolism enzyme activation effects. Meanwhile, after extensive searching, no existing technologies have been found to apply the peptide with the amino acid sequence Pro-Phe-Tyr-Pro (PFYP) to hangover relief and liver protection products, especially not reports of its use in combination with curcumin and puerarin using the aforementioned specific delivery system.

[0006] This invention addresses the specific need for "oral hangover relief and liver protection" by proposing a systematic solution: First, precise formulation, using curcumin (antioxidant), puerarin (promotes alcohol metabolism), and the validated novel hangover-relieving peptide PFYP to directly target multiple stages of hangover relief; Second, innovative delivery, employing a three-step process of "encapsulation-double emulsification-gelation" to construct a double emulsification system and immobilize it within calcium alginate gel microbeads. This structure effectively resists gastric acid and enzymatic degradation, providing comprehensive protection for the three components with different physicochemical properties (especially the easily inactivated PFYP peptide), while simultaneously addressing the differences in hydrophilicity and hydrophobicity between the wall material and the active ingredient, and achieving sustained intestinal release; Third, synergistic effect, this design ensures the synergistic release of each component in both time and space, covering the entire process from alcohol metabolism to liver damage repair, and achieving a synergistic effect of "1+1+1>3" (where 1+1+1>3). Figure 8 ).

[0007] In view of this, the present invention is proposed. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a composite gel microbead with hangover relief and liver protection functions, as well as its preparation method. These microbeads can effectively improve the solubility and stability of poorly soluble components, protect active peptides from pepsin degradation, and achieve synergistic sustained release of multiple components through dual emulsification and gelation technologies, thereby enhancing bioavailability and overall efficacy.

[0009] To achieve the above objectives, the technical solution of the present invention is as follows:

[0010] A method for preparing composite gel microbeads with hangover relief and liver protection functions, characterized by comprising the following steps: 1. Preparation of curcumin-encapsulated lyophilized powder: Curcumin was mixed with an aqueous solution of 2-hydroxypropyl-β-cyclodextrin to carry out an inclusion reaction. After the reaction was completed, the mixture was centrifuged, and the supernatant was freeze-dried to obtain the curcumin-cyclodextrin inclusion complex lyophilized powder. 2. Preparation of puerarin oil phase: Puerarin was dissolved in corn oil, and lecithin was added as an emulsifier. The mixture was heated and stirred until completely dissolved and mixed evenly to obtain a clear oil phase. 3. Preparation of the aqueous phase of the novel active peptide: The lyophilized powder obtained in step 1 and a specific peptide with hangover-relieving function are dissolved together in an aqueous solution and mixed evenly to obtain an aqueous phase; the peptide is a novel hangover-relieving active peptide prepared by enzymatic hydrolysis, identified by sequencing, chemically synthesized, and verified for activity. 4. Preparation of primary emulsion: The aqueous phase obtained in step 3 is mixed with the oil phase obtained in step 2 and homogenized at high speed to form a primary emulsion. 5. Formation of gel microspheres: The primary emulsion obtained in step 4 is premixed with sodium alginate solution and then homogenized and emulsified again to form a uniform mixed emulsion. This mixed emulsion is then dropped into calcium chloride solution, where it crosslinks and solidifies to form gel microspheres. The microspheres are collected and washed with deionized water to obtain the composite gel microspheres.

[0011] Preferably, in step 1, the molar ratio of curcumin to hydroxypropyl-β-cyclodextrin is 1:1 to 1:6; the freeze-drying temperature is -40°C to -80°C, and the vacuum degree is less than 10 Pa.

[0012] Preferably, in step 2, the amount of lecithin added is 1%-5% (w:v) of the volume of corn oil.

[0013] Preferably, in step 3, the sequence of the novel alcohol-detoxifying active peptide is a short peptide that has been prepared by enzymatic hydrolysis, sequencing and chemical synthesis, and verified by cell experimental models to have the activation function of alcohol dehydrogenase and acetaldehyde dehydrogenase, and its amino acid sequence is PFYP (Pro-Phe-Tyr-Pro).

[0014] Preferably, in step 4, the volume ratio of the aqueous phase to the oil phase is 1:1 to 1:4.

[0015] Preferably, in step 4, the homogenization speed is 8000-12000 rpm and the time is 1-3 min.

[0016] Preferably, in step 5, the concentration of sodium alginate solution is 1%-3%; the concentration of calcium chloride solution is 1%-5%; and the crosslinking time is 10-60 minutes.

[0017] This invention also claims protection for the composite gel microbeads obtained by the above method.

[0018] This invention also claims protection for the use of the composite gel microbeads in the preparation of food, health products or pharmaceuticals with hangover relief and / or liver protection functions.

[0019] Compared with the prior art, the present invention has the following beneficial effects: (1) Synergistic effect of multiple components: This invention integrates curcumin, puerarin and specific novel active peptides to produce a synergistic effect on the entire chain of "metabolism-damage-repair". The three components achieve dynamic coordination of functions through composite gel microbeads, and the overall effect is significantly better than that of single components or physical mixtures. (2) Significantly improve bioavailability and stability: This invention adopts a step-by-step delivery strategy to overcome the difficulties of each component in application. After curcumin is encapsulated by cyclodextrin, its water solubility and stability are improved. Puerarin is absorbed through the lymphatic system with the help of lipid carriers, effectively avoiding the first-pass effect and increasing the concentration of the original drug in the whole body and liver targets. The active peptide is protected by the outer gel layer, which can resist gastrointestinal degradation and ensure active delivery. (3) Achieving programmed protection and synergistic sustained release: The "aqueous phase / oil phase / hydrogel" triple delivery system constructed in this invention achieves programmed delivery of active ingredients in the digestive tract. The outer calcium alginate gel network can effectively reduce the burst release of active ingredients in the acidic environment of the stomach and provide a physical barrier, significantly protecting novel peptides, especially those that are easily degraded, from pepsin hydrolysis; after entering the neutral environment of the intestine, the gel network gradually dissolves, and the internal droplets are released. (4) Controllable process and uniform and reliable product: This invention adopts a step-by-step process of "encapsulation-emulsification-gelation" to prepare freeze-dried powder containing curcumin in advance to ensure dispersion stability; a uniform emulsion is formed first to prevent phase separation; and mild ionic cross-linking is used to protect the activity. The process is easy to scale up, and the resulting microbeads have high encapsulation efficiency and uniform particle size, ensuring stable and reproducible efficacy. Attached Figure Description

[0020] Figure 1 This is a flowchart of the preparation process for novel peptides.

[0021] Figure 2 These are flowcharts of the preparation process of composite gel microbeads and intestinal digestion diagrams.

[0022] Figure 3 This is an HPLC chromatogram of the synthesized novel peptide.

[0023] Figure 4 This is a schematic diagram of a dual emulsification system.

[0024] Figure 5 This is a schematic diagram of composite gel microbeads loaded with multiple active ingredients.

[0025] Figure 6 This is a schematic diagram of simulated digestion and release of composite gel microbeads.

[0026] Figure 7 This is the infrared spectrum of composite gel microbeads loaded with multiple active ingredients.

[0027] Figure 8 This is a graph showing the activation rate of alcohol dehydrogenase in three single components and composite gel microbeads.

[0028] Figure 9 This is a graph showing the aspartate aminotransferase (AST) activity results of three single components and composite gel microbeads. Detailed Implementation

[0029] It should be understood that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. To enable those skilled in the art to more clearly understand the technical solutions of the present invention, the technical solutions of the present invention will be described in detail below with reference to specific embodiments. The experimental materials used in the embodiments of the present invention are all conventional experimental materials in the art and can be purchased through commercial channels.

[0030] The first embodiment of this invention provides a method and process for preparing the discovered novel peptides. First, a mixture containing multiple peptides is prepared by enzymatic hydrolysis. This mixture is then separated and purified using a Sephadex G-15 dextran gel column, and absorbance-different collection liquid curves are generated to obtain different components. The optimal components are screened by detecting the enzyme activities of alanine aminotransferase and aspartate aminotransferase. The separation results are identified by LC-MS / MS technology, and the amino acid sequences are obtained by database retrieval. The activity of peptides with different amino acid sequences is screened using bioinformatics. Simultaneously, the obtained peptides are docked with two receptor proteins, ADH1B and ALDH2, using Autodock software. Peptides with the strongest binding affinity to ADH1B and ALDH2 are selected based on binding energy. The screened peptides are synthesized using a solid-phase synthesis method, and their synthetic purity is identified by chromatography. Their activity is verified by detecting the activation rates of alcohol dehydrogenase and acetaldehyde dehydrogenase.

[0031] The second embodiment of this invention provides a method for preparing composite gel microbeads loaded with multiple active ingredients. First, curcumin and 2-hydroxypropyl-β-cyclodextrin are mixed in an aqueous solution for an inclusion reaction. After 4 hours of reaction, the mixture is centrifuged, and the supernatant is freeze-dried to obtain a curcumin-cyclodextrin inclusion complex lyophilized powder. The obtained lyophilized powder is dissolved together with the specific peptide fragment with hangover-relieving function obtained above in an aqueous solution and mixed evenly to obtain an aqueous phase. Puerarin is dissolved in corn oil, and lecithin is added as an emulsifier. The mixture is heated and stirred until completely dissolved and mixed evenly to obtain a clear oil phase. The aqueous phase and oil phase are mixed and homogenized at high speed to form a primary emulsion. This primary emulsion is then premixed with sodium alginate solution and homogenized again to form a homogeneous mixed emulsion. This mixed emulsion is dropped into a calcium chloride solution for cross-linking and solidification to form gel microbeads. The microbeads are collected and washed with deionized water to obtain the composite gel microbeads.

[0032] To make the technical solution of the present invention clearer, the following detailed description is provided through several specific embodiments.

[0033] Example 1: Preparation of Novel Peptides (1) Preparation of enzyme hydrolysate Take 10g of corn gluten powder, add 100mL of water, heat to dissolve at 50℃, cool to room temperature after high-temperature pretreatment, adjust pH to 7, add protease after preheating, enzymatic hydrolysis at 55℃ for 2 hours, inactivate enzyme at 95℃ for 20 minutes, centrifuge at 4000rpm for 20 minutes, take the supernatant for ultrafiltration, place in a 100Da dialysis bag and dialyze in deionized water for 24 hours, freeze dry to obtain corn peptides. (2) Isolation and purification of corn peptides Sephadex G-15 gel particles were soaked overnight at room temperature until fully swollen. The gel was then packed into a 1.6 × 80 cm glass chromatography column, and eluted with ultrapure water to reach column equilibration. A lyophilized corn peptide solution was prepared at 20 mg / mL, filtered through a 0.22 μm filter, and eluted with ultrapure water at a flow rate of 1 mL / min, collecting 5 mL samples every 5 min. The eluent was analyzed at 220 nm using a UV-Vis spectrophotometer, and peak shapes were plotted. Peaks from the same elution peak were combined and lyophilized for later use. (3) Amino acid sequence identification and bioinformatics screening of corn peptides The sample was treated with ultrapure water, followed by reductive alkylation and acid adjustment. Desalting was performed using a C18 StageTip, and the sample was vacuum-dried at 45°C before LC-MS / MS analysis to identify the amino acid sequence. Bioinformatics screening was conducted using databases. The obtained peptides were docked with the ADH1B and ALDH2 receptor proteins using Autodock software. Lower binding energies indicated stronger affinity and a tighter binding between the two molecules. The bioinformatics results were then used to screen peptides. (4) Synthesis and activity verification of corn peptides Polypeptides were synthesized using a solid-phase synthesis method, and their activity was verified through cell experiments. The specific procedures were as follows: HepG2 cells were cultured at 3.0*10⁻⁶ cells / cells. 5 Cells were seeded at a density of [number] cells / mL in 6-well plates. After 24 h of culture, the sample group was treated with 1 mg / mL of sample solution, while the control and model groups were treated with the same amount of culture medium. After another 24 h of culture, the sample and model groups were treated with ethanol solution, while the control group was treated with the same amount of culture medium. The plates were then incubated for 24 h. Subsequently, RNA was extracted using Trizol reagent and cDNA was formed by reverse transcription. The relative expression levels of alcohol dehydrogenase and aldehyde dehydrogenase were measured using real-time quantitative PCR to verify that corn peptide enhances the activity of these two alcohol-metabolizing enzymes. The purity of the synthesized peptide is determined by HPLC. The purity can be calculated by determining the ratio of the main peak area to the total peak area. Figure 3 It can be seen that the peak elution time of the synthesized peptide was 9.675 min, and the purity was identified as 98.072%.

[0034] Example 2: Preparation of composite gel microbeads loaded with multiple active ingredients (1) Preparation of freeze-dried powder containing curcumin Weigh 1.5 g of 2-hydroxypropyl-β-cyclodextrin and dissolve it in 15 mL of pure water; dissolve 100 mg of curcumin in 10 mL of anhydrous ethanol to prepare a 10 mg / mL solution. Under magnetic stirring at 60 °C, slowly add the curcumin solution dropwise to the 2-hydroxypropyl-β-cyclodextrin solution, and allow the inclusion reaction to proceed in the dark for 4 hours. After cooling, centrifuge at 12000 rpm for 15 minutes, collect the supernatant, pre-freeze at -80 °C, and then freeze-dry under a vacuum of 5 Pa for 48 hours to obtain curcumin-encapsulated lyophilized powder. Dissolve the lyophilized powder in water, and determine the encapsulation efficiency of curcumin using solvent extraction-HPLC. (2) Preparation of dual emulsification system Weigh 200 mg of puerarin and dissolve it in 30 mL of corn oil. Add 100 mg of lecithin and stir in a 50 °C water bath until homogeneous to obtain the oil phase. Weigh 100 mg of a novel hangover-relieving peptide (sequence: Pro-Phe-Tyr-Pro) and 150 mg of curcumin-encapsulated lyophilized powder and dissolve them in 10 mL of phosphate buffer to obtain the aqueous phase. Mix the aqueous and oil phases and homogenize at 10,000 rpm for 2 min to obtain the primary emulsion. The primary emulsion was premixed with 50 mL of 2% sodium alginate solution and homogenized again at 8000 rpm for 50 s to obtain a pale yellow emulsion, such as... Figure 4 As shown. (3) Preparation of composite gel microbeads and determination of the encapsulation efficiency of three active ingredients The mixed emulsion was added dropwise to a 2% calcium chloride solution using a 2mL syringe, and cross-linked for 30 minutes. The microbeads were collected by filtration and washed three times with pure water to obtain pale yellow gel microbeads. Figure 5 As shown. The encapsulation efficiency of curcumin, puerarin, and peptides in composite gel microbeads was determined using a combined extraction and chromatography (PEC) scheme. Appropriate amounts of gel microbeads were weighed and ground. High-efficiency extraction was performed using a methanol-water solution containing a small amount of trifluoroacetic acid, via vortexing and ice-bath sonication. After centrifugation, the supernatants were combined to obtain the sample solution. Simultaneous quantitative analysis was performed using reversed-phase high-performance liquid chromatography (RP-HPLC). A gradient elution program enabled effective separation of the three components in a single chromatographic run. Data were acquired using a diode array detector at characteristic wavelengths of 220 nm (peptide), 250 nm (puerarin), and 430 nm (curcumin), and the content was calculated based on their respective standard curves. Finally, the encapsulation efficiency and drug loading of each component were calculated using the following formula. The encapsulation rates of curcumin-encapsulated lyophilized powder in the composite gel microbeads were 81.62±0.19%, puerarin 78.59±0.37%, and novel peptides 90.17±0.62%. This indicates that the three active ingredients loaded onto the composite gel microbeads all have high loading rates and good performance.

[0035] Example 3 Characterization of composite gel microbeads loaded with multiple active ingredients Take appropriate amounts of curcumin, curcumin-encapsulated lyophilized powder, puerarin, novel peptides, and gel microbeads obtained in Example 2, and determine the particle size and Fourier transform infrared spectrum (FT-IR). (1) Particle size determination Each sample was uniformly dispersed in ultrapure water. 1 mL of each suspension was taken and measured using a nanoparticle size analyzer and a zeta potential analyzer. The temperature was set at 25℃, and the measurement angle was 90 degrees. The results are shown in the table below. (2) Fourier transform infrared spectroscopy determination The prepared gel microbeads were freeze-dried and ground, then mixed with potassium bromide at a ratio of 1:100. After thorough grinding, the mixture was compressed into tablets. Fourier transform infrared spectroscopy (FTIR) was used to analyze different samples, with the scanning range set at 4000-4000 cm⁻¹. -1 The infrared absorption spectrum was obtained by scanning 32 times. Depend on Figure 7 It can be seen that the composite gel microspheres are 3300 cm² -1 The weakening of the hydrogen bond peaks around 2851 cm⁻¹ indicates that the water-centered hydrogen bond network of the freeze-dried gel microspheres was completely removed. -1 and 1746cm -1 New peaks appeared at the microspheres, namely the symmetric stretching vibration peak of CH2 in the fatty chain and the characteristic peak of C=O stretching vibration of aliphatic esters. This indicates that the main components of corn oil are tightly attached to the surface or internal pore walls of the microspheres, and the lipid phase (corn oil) is exposed and concentrated. In the composite gel microspheres, puerarin and curcumin are embedded lyophilized powders at 800-900 cm⁻¹. -1 The absorption peaks between [a certain point] disappeared, and the peptide peak at 850 cm⁻¹ disappeared. -1 The absorption peaks around the left and right disappear; these peaks are typically due to the out-of-plane bending vibrations of the CH bonds on the aromatic ring. In the final lyophilized microbeads, these active molecules are deeply embedded, first physically encapsulated by a cyclodextrin / peptide / sodium alginate network, then immobilized by a calcium ion-crosslinked gel network, and finally interacting with lipids during lyophilization. This embedding and confinement leads to the suppression of local vibrational modes of the aromatic ring, resulting in significant broadening and attenuation of its characteristic peaks, thus manifesting as the disappearance of absorption peaks in the spectrum, indicating that the active ingredient has been successfully embedded.

[0036] Determination of the activation rate of alcohol-metabolizing enzymes in a comparative manner This application example uses the Waller-Hoch method to determine the alcohol dehydrogenase activation rate of the composite gel microbeads and three single active ingredients prepared in Example 2. The results are as follows: Figure 8 As shown, the alcohol dehydrogenase activation rates of curcumin-encapsulated lyophilized powder, puerarin, and novel peptides were 24.36±1.17%, 41.10±1.51%, and 72.24±0.96%, respectively. The alcohol dehydrogenase activation rate of the composite gel microbeads prepared in Example 2 was 151.33±1.19%. This indicates that the combination of puerarin, curcumin, and novel peptides achieved a synergistic effect of "1+1+1>3", effectively improving the alcohol dehydrogenase activation rate. Meanwhile, compared with current research, for example, the activation rate of ADH in the beverage with hangover-relieving and liver-protecting effects in the authorized patent "A composition for relieving hangovers and protecting liver, and its preparation method" is around 30%; the ADH activity in mice in the authorized patent "A composition for relieving hangover headaches and its preparation method" is about 80%-100% higher than that in the blank control; and the ADH level in the liver of rats in the complex group in the currently published paper "A Herbal Composition of Semen Hoveniae, Radix Puerariae. and Fructus Schisandrae Shows Potent Protective Effects on Acute Alcoholic Intoxication in Rodent Models" is about 40% higher than that in the control group. This indicates that the composite gel microbeads prepared in this invention have superior activation of alcohol-metabolizing enzymes, thereby achieving the purpose of relieving hangovers and protecting the liver. Simultaneously, the effects of the composite gel microbeads and three single active ingredients prepared in Example 2 on the activity of aspartate aminotransferase (AST) in alcohol-damaged hepatocytes were determined using an AST kit. Figure 9 As shown, the composite gel microbeads reduced AST activity to the greatest extent, decreasing it from 40.53±0.83 U / gprot in the model group to 12.49±1.13 U / gprot, indicating that the prepared composite gel microbeads have a good effect on relieving hangovers and protecting the liver.

[0037] In summary, the technical solution of this invention prepares novel composite gel microbeads with hangover relief and liver protection functions by double emulsifying an innovative novel peptide segment with active ingredients of different physicochemical properties, thereby improving the bioavailability and stability of the active ingredients.

[0038] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A method for preparing composite gel microbeads with hangover relief and liver protection functions, characterized in that, The method comprises the following steps: a) Inclusion complex preparation: mixing curcumin and 2-hydroxypropyl-β-cyclodextrin in an aqueous solution to perform a molecular inclusion reaction, then centrifuging the inclusion reaction solution and taking the supernatant to perform freeze-drying, thereby obtaining a freeze-dried powder of curcumin-2-hydroxypropyl-β-cyclodextrin inclusion complex; b) Lipid oil phase construction: completely dissolving puerarin in corn oil, and adding lecithin as an emulsifier to uniformly disperse and form a stable puerarin lipid oil phase; c) Active water phase construction: dissolving the inclusion complex freeze-dried powder obtained in step a) and a novel peptide segment with alcohol-protecting and liver-protecting activity in an aqueous medium, and uniformly mixing to obtain a complex water phase containing curcumin inclusion complex and active peptides; d) Primary emulsification: mixing the complex water phase obtained in step c) and the puerarin lipid oil phase obtained in step b) at a volume ratio of 1:1 to 1:4, performing first emulsification by high-speed homogenization to form a pre-dispersion system for the next emulsification; e) Double emulsification and gelation: pre-mixing the primary emulsion obtained in step d) with a sodium alginate aqueous solution, and then performing second emulsification by homogenization treatment to form a uniform double emulsification system; f) Introducing the double emulsification system obtained in step e) into a calcium chloride solution by means of syringe dropwise addition, and using the ionic crosslinking effect of sodium alginate and calcium ions to solidify and form a complex gel microbead loaded with curcumin inclusion complex, puerarin lipid carrier and active peptides.

2. The production method according to claim 1, characterized by, In step a), the molar ratio of curcumin to 2-hydroxypropyl-β-cyclodextrin is 1:1 to 1:

6.

3. The preparation method according to claim 1, characterized in that, In step c), the novel peptide segment with alcohol-protecting and liver-protecting activity is a bioactive peptide prepared by enzymatic preparation, separation and sequencing, chemical synthesis, and verified by in vitro ethanol dehydrogenase activation experiment to have significant alcohol-protecting activity, and the amino acid sequence is Pro-Phe-Tyr-Pro (PFYP).

4. The production method according to claim 1, characterized by, In step d), the first emulsification adopts high-speed dispersion homogenization, the rotation speed is 8000-12000 rpm, and the treatment time is 1-5 minutes.

5. The method of claim 1, wherein, In step e), the mass concentration of the sodium alginate aqueous solution is 1% to 4%, the mass concentration of the calcium chloride solution is 1% to 5%, and the ionic crosslinking solidification time is 10-60 minutes.

6. The production method according to claim 1, characterized by, In step f), the structure of the complex gel microbead formed is characterized in that: The inner layer emulsion droplet is the dispersed phase, and the outer layer is coated and fixed by a sodium alginate-calcium ion crosslinked gel network, thereby forming a core-shell type double emulsification gel delivery system.

7. A complex gel microbead prepared by the preparation method of any one of claims 1 to 6.

8. The composite gel microbead of claim 7, wherein, It comprises a gel network formed by sodium alginate-calcium ion crosslinking, and a plurality of independent droplets encapsulated in the gel network; the oil phase in the independent droplets is corn oil containing puerarin and lecithin, the water phase contains curcumin-2-hydroxypropyl-β-cyclodextrin inclusion complex and active peptides with the amino acid sequence Pro-Phe-Tyr-Pro; and the gel matrix can provide protection in the gastrointestinal environment to achieve sustained release of active ingredients.

9. Use of the composite gel microbeads according to claim 7 or 8 for the preparation of a food, a health food or a drug for preventing or alleviating symptoms of drunkenness, reducing the risk of alcoholic liver damage.

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