A complex peptide nutrient having a fat-reducing effect

Abstract

This invention discloses a compound peptide nutrient with fat-reducing effects, belonging to the field of health food technology. Specifically, it relates to a VQPIPY peptide or a product containing VQPIPY peptide in the preparation of lipid-lowering drugs or health products, or in the preparation of drugs for treating and / or alleviating alcoholic liver damage. This invention also provides a meal replacement powder containing VQPIPY peptide, shown as a compound of whey powder, milk powder, oats, spinach powder, allulose, and camel milk peptide in a specific ratio. Through the synergistic effect of its components, this product achieves multiple effects such as providing satiety, inhibiting blood sugar, and lowering blood lipids. It is characterized by balanced nutrition, safety, effectiveness, and good taste, and can be used for weight management and daily health maintenance.

Description

Technical Field

[0001] This invention belongs to the field of health food technology, specifically relating to a compound peptide nutrient with fat-reducing effects. Background Technology

[0002] With the improvement of living standards, obesity and its related metabolic diseases (such as hyperlipidemia and diabetes) have become a global health problem. Currently, there are various weight-loss products on the market, mainly including chemically synthesized drugs, meal replacement foods, and health supplements. While chemical drugs are fast-acting, they often have significant side effects and are not suitable for long-term use. Many meal replacement foods or health supplements have limited functions, either only providing a feeling of fullness without nutritional balance, or focusing only on one aspect of metabolic regulation, resulting in limited effectiveness and poor taste, leading to poor user compliance. Therefore, there is an urgent need in this field for a comprehensive weight management product that combines safety, effectiveness, and good taste.

[0003] Food protein bioactive peptides are rich in nutritional value and possess various physiological functions such as antioxidation, anti-fatigue, and immune regulation, leading to their widespread application in food. They hold promise as a potential replacement for nutrients such as proteins and amino acids in the future. Summary of the Invention

[0004] The purpose of this invention is to screen and prepare an active peptide with strong lipid-lowering activity, and to prepare a compound nutritional meal replacement powder.

[0005] This invention provides the use of VQPIPY peptide or products containing VQPIPY peptide in the preparation of lipid-lowering drugs or health products, or in the preparation of drugs for treating and / or alleviating alcoholic liver injury, wherein the amino acid sequence of the VQPIPY peptide is shown in SEQ ID NO.1 (Val-Gln-Pro-Ile-Pro-Tyr).

[0006] In one embodiment of the present invention, the product containing VQPIPY peptide is a meal replacement powder containing VQPIPY peptide.

[0007] In one embodiment of the present invention, the meal replacement powder is formulated as follows: 20-25 parts whey powder, 15-20 parts milk powder, 15-25 parts oats, 5-15 parts spinach powder, 5-15 parts allulose, 15-25 parts coix seed powder, 0.5-2 parts white kidney bean extract, 1-5 parts inulin, and 1-2 parts VQPIPY peptide.

[0008] In one embodiment of the present invention, the pharmaceutical product further contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field.

[0009] In one embodiment of the present invention, the excipients include one or more of the following: binders such as cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, and mannitol; fillers such as starch and sucrose; humectants such as glycerin; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, fatty acid sorbitan, and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, and pharmaceutical iron oxide red; lubricants such as hydrogenated vegetable oil, talc, and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, and cellulose acetate; and other excipients such as flavoring agents and sweeteners may also be added to the composition.

[0010] In one embodiment of the present invention, the dosage form of the drug includes, but is not limited to, oral dosage form, injection dosage form, and inhalation dosage form.

[0011] In one embodiment of the present invention, the oral dosage form includes, but is not limited to, tablets, capsules, granules, oral liquids, and oral suspensions; In one embodiment of the present invention, the injectable dosage form includes, but is not limited to, injectable liquid and injectable powder.

[0012] In one embodiment of the present invention, the inhalation dosage form includes, but is not limited to, aerosols and powder inhalers.

[0013] In one embodiment of the present invention, the health product also contains acceptable excipients.

[0014] The present invention also provides a drug or health product for lowering blood lipids, or a drug for treating and / or alleviating alcoholic liver damage, wherein the active ingredient is the active peptide VQPIPY peptide.

[0015] In one embodiment of the present invention, the pharmaceutical product further contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field.

[0016] In one embodiment of the present invention, the excipients include one or more of the following: binders such as cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, and mannitol; fillers such as starch and sucrose; humectants such as glycerin; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, fatty acid sorbitan, and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, and pharmaceutical iron oxide red; lubricants such as hydrogenated vegetable oil, talc, and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, and cellulose acetate; and other excipients such as flavoring agents and sweeteners may also be added to the composition.

[0017] In one embodiment of the present invention, the dosage form of the drug includes, but is not limited to, oral dosage form, injection dosage form, and inhalation dosage form.

[0018] In one embodiment of the present invention, the oral dosage form includes, but is not limited to, tablets, capsules, granules, oral liquids, and oral suspensions; In one embodiment of the present invention, the injectable dosage form includes, but is not limited to, an injection solution or an injection powder for injection; In one embodiment of the present invention, the inhalation dosage form includes, but is not limited to, aerosols and powder inhalers.

[0019] In one embodiment of the present invention, the health product further contains acceptable excipients. The present invention provides a method for preparing camel milk active peptides and meal replacement powder. Camel milk peptides are used as the main raw material, and camel milk protein polypeptides are prepared by bromelain hydrolysis. After purification, purified peptides are obtained and used to produce meal replacement powder.

[0020] This study used camel milk as a substrate and employed bromelain for enzymatic hydrolysis. Specific hydrolysis conditions were as follows: substrate concentration 1040 g / L, enzyme dosage 500-10000 U / g, hydrolysis temperature 35-55 ℃, hydrolysis time 1-5 h, and hydrolysis pH 5.5-7.5. The hydrolysates were further separated using a Sephadex G-15 dextran gel column. The column was pre-equilibrated with Tris-acetate buffer (pH 7.0), followed by elution with pure water at a flow rate of 0.5 mL / min. UV absorption was monitored at 280 nm, and the eluted fractions were collected based on the absorption peaks. Lipase inhibitory activity was determined for each fraction. The protein concentrations of fractions M1, M2, and M3 were 7.9 mg / mL, 23.6 mg / mL, and 15.1 mg / mL, respectively. Activity assessment showed that fraction M2 exhibited good lipase inhibitory activity, with an IC50 value of [missing value]. 50 The value was 0.34 mg / mL, so it was named Camel Milk Active Peptide M2.

[0021] For further purification, the M2 fraction was prepared as a 20 mg / mL aqueous solution and separated by preparative reversed-phase high-performance liquid chromatography (RP-HPLC). A C18 column was used, with a mobile phase of 2-85% (v / v) methanol containing 0.02-0.15% (v / v) trifluoroacetic acid at a flow rate of 0.5 mL / min. The fraction was detected at 280 nm and collected in conjunction with mass spectrometry. After 4-6 rounds of repeated purification, high-purity fraction K3 was finally obtained, with a peptide purity of 95.6 g / 100 g and significantly improved lipase inhibitory activity. 50The value was 0.08 mg / mL. LC-ESI-Orbitrap MS / MS analysis revealed that the major peptide sequence in the K3 fraction was VQPIPY, with an abundance of 88.67% in the sample.

[0022] This invention provides a method for preparing camel milk peptide meal replacement powder, which is composed of the following raw materials by weight percentage: whey powder 15%-25%, milk powder 10%-20%, oats 20%-30%, spinach powder 3%-15%, allulose 5%-10%, coix seed powder 20%-30%, white kidney bean extract 0-16%, inulin 0-10%, and active peptide M2 ​​added at 1%-3%.

[0023] Preferably, the whey powder is whey protein concentrate or whey protein isolate; the milk powder is whole milk powder or skim milk powder; and the oats are pregelatinized oat flour.

[0024] This invention provides a method for preparing the above-mentioned compound nutritional meal replacement powder, comprising the following steps: Raw material pretreatment: Whey powder, milk powder, oat powder, Job's tears powder, spinach powder, etc. are passed through an 80-100 mesh sieve to ensure uniform powder fineness.

[0025] Weighing and mixing: Accurately weigh each raw material according to the above ratio, and gradually mix the trace components with the large amount of base powder using the equal increment method.

[0026] Final mixing and sterilization: Place the pre-mixed materials into a three-dimensional motion mixer and mix for 30-45 minutes to ensure complete homogeneity. Sterilize using cobalt-60 irradiation or microwave low-temperature sterilization.

[0027] Packaging: The finished product is obtained by quantitative dispensing and sealing in a clean room.

[0028] The present invention provides the application of the compound nutritional meal replacement powder in the preparation of health food for weight management, improving intestinal health and providing comprehensive nutrition.

[0029] This invention also provides a peptide and weight-loss compound nutritional meal replacement powder with lipid-lowering effects, using camel milk peptide as the main raw material, and preparing camel milk protein polypeptide by bromelain hydrolysis, and then purifying the peptide to make meal replacement powder.

[0030] Preferably, camel milk is used as the substrate, and bromelain is selected for enzymatic hydrolysis; the substrate concentration is 1040 g / L, the enzyme addition amount is 500-10000 U / g, the hydrolysis temperature is 35-55 ℃, the hydrolysis time is 1-5 h, and the hydrolysis pH is 5.5-7.5.

[0031] Camel milk protein hydrolysates were purified by Sephadex column chromatography using dextran gel. The column was equilibrated with Tris-acetate buffer (pH 7.0), eluted with water at a flow rate of 0.5 mL / min, and fractions were collected based on UV absorption (280 nm). The lipase inhibitory activity of different fractions was determined. The protein concentrations of fractions M1, M2, and M3 were 7.9 mg / mL, 23.6 mg / mL, and 15.1 mg / mL, respectively. Fraction M2 showed better lipase inhibitory activity, with an IC50 value of 0.34 mg / mL.

[0032] The M2 component was further separated and purified. The sample solution was injected into a preparative liquid chromatography reverse-phase column (C18) with a mobile phase of 2-85% (v / v) methanol containing 0.02-0.15% (v / v) trifluoroacetic acid. The eluent was collected based on UV absorption intensity. The component with the best lipase inhibitory activity was obtained and then purified again using the same method. Through 4-6 purification cycles, a high-purity active component K3 was finally obtained. The peptide content (purity) of K3 reached 95.6 g / 100 g. The K3 component exhibited strong lipase inhibitory activity, with an IC50 value of 0.08 mg / mL. LC-ESI-Orbitrap MS / MS identification showed that the major peptide sequence in the K3 component was VQPIPY, with an abundance of 88.67% in the sample.

[0033] The present invention also provides a method for preparing camel milk peptide and its meal replacement powder, which is composed of the following raw materials in weight percentage: whey powder 15%-25%, milk powder 10%-20%, oats 10%-15%, spinach powder 3%-6%, allulose 5%-10%, coix seed powder 5%-10%, white kidney bean extract 0-8%, inulin 0-10%, and active peptide M2 ​​added at 1%-3%. In one embodiment of the present invention, the whey powder is whey protein concentrate or whey protein isolate.

[0034] In one embodiment of the present invention, the milk powder is skimmed milk powder.

[0035] The present invention also provides a method for preparing the above-mentioned compound nutritional meal replacement powder, comprising the steps of: raw material pretreatment, weighing and mixing, total mixing and sterilization, and packaging.

[0036] The present invention also provides the application of the above-mentioned compound nutritional meal replacement powder in the preparation of health food for weight management.

[0037] This invention also provides the application of the above-mentioned compound nutritional meal replacement powder in the preparation of health food for improving intestinal health.

[0038] Beneficial effects (1) The present invention has good effects and high safety: the prepared active peptides have strong lipid-lowering activity, and all components are derived from natural foods or traditional medicinal and food herbs, avoiding the side effects of chemical drugs and suitable for long-term use.

[0039] (2) The meal replacement powder of the present invention has good taste and compliance: by improving the flavor and sweetness through whey powder, milk powder, allulose, etc., the unpleasant taste of herbs is masked, which greatly increases the user's willingness to continue using it.

[0040] (3) The peptides of the present invention have significant weight control effects, and the peptides obtained by the present invention have significant liver protection, lipid-lowering effects and antioxidant activity, showing promising applications in these aspects. Furthermore, they can better exert the effects of lipid-lowering and weight control. Detailed Implementation

[0041] The present invention will be further described below with reference to embodiments and comparative embodiments, but the scope of protection of the present invention is not limited thereto. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0042] The basal feed and high-fat feed involved in the following examples were purchased from Beijing Boaigang Biotechnology Co., Ltd. The high-fat feed formula added 20% lard, 1.2% cholesterol and 0.2% sodium cholate to the basal feed.

[0043] The detection methods involved in the following embodiments are as follows: Pancreatic lipase activity assay Prepare PBS with a pH of 7.0 and a concentration of 50 mmol / L for later use.

[0044] Accurately weigh p -NP 94.5941 mg, dissolved in 100 mL N , N Dimethylformamide was used to obtain a stock solution of 3.4 mmol / L, which was then diluted to 0, 0.43, 0.85, 1.70, and 2.55 mmol / L. 500 μL of PBS, 1000 μL of 10% DMSO (v / v), and 200 μL of different concentrations were added. p -NP solution is mixed evenly to make the final p -NP concentrations were 0, 0.05, 0.10, 0.20, and 0.40 mmol / L. 200 μL was added to each well of a 96-well plate, and the absorbance was measured at 405 nm. p Plot a graph with NP concentration (mmol / L) on the x-axis and absorbance on the y-axis. p- The absorbance standard curve of NP was fitted to obtain a linear equation.

[0045] The method for determining PL activity uses spectrophotometry. p -NPB was used as the substrate. 20 mg of PL was weighed and added to an appropriate amount of PBS, then diluted to a 10 mL volumetric flask. The flask was then centrifuged at 8000 r / min for 15 min at 10 °C. The supernatant was collected to obtain a 2 mg / mL PL solution. 10 μL of this solution was then... p Add an appropriate amount of NPB N , N Dimethylformamide was added and diluted to a 10 mL volumetric flask to obtain a 6 mmol / L substrate solution. 500 μL of PL solution and 1000 μL of 10% DMSO solution (v / v) were thoroughly mixed in a centrifuge tube and incubated at 37 °C for 1 h. Then, 200 μL of substrate solution was added, and the mixture was shaken well and incubated for another 20 min. The reaction was terminated in a boiling water bath, cooled, and centrifuged at 10000 r / min for 10 min at 25 °C. 200 μL of the supernatant was quickly spotted onto a TLC plate, and the absorbance at 405 nm was measured. p Substrate content is calculated using the NP absorbance standard curve. Enzyme activity is calculated using the following formula (2-1): (2-1) In the formula: enzyme activity, U / g; n is the dilution factor; V is the volume of the sample solution to be tested, mL; C is the amount of enzyme activity carried over to the target sample. p -NP standard curve calculation p -NP concentration, mmol / L; t is the reaction time, min; 0.001 is the mass of PL in the reaction system, g.

[0046] Example 1: Preparation method of bioactive peptides This study used camel milk as a substrate and employed bromelain for enzymatic hydrolysis.

[0047] The specific enzymatic hydrolysis conditions are as follows: (1) Add bromelain (2000 U / g) to camel milk (1040 g / L substrate concentration) and react at 50℃ and pH 7.0 for 4 hours to obtain the enzymatic hydrolysis product.

[0048] (2) The enzymatic hydrolysis products were further separated by Sephadex G-15 dextran gel column: the column was pre-equilibrated with Tris-acetate buffer (pH 7.0), and then eluted with pure water at a flow rate of 0.5 mL / min. The UV absorption was monitored at a wavelength of 280 nm, and the eluted components were collected according to the absorption peak. The M1, M2 and M3 components were obtained at elution times of 95, 171 and 236 min, respectively.

[0049] (3) The lipase inhibitory activity of each component was determined. The protein concentrations of components M1, M2, and M3 were 7.9 mg / mL, 23.6 mg / mL, and 15.1 mg / mL, respectively. Activity assessment showed that component M2 had better lipase inhibitory activity, with an IC50 value of [missing value]. 50 The concentration was 0.34 mg / mL, therefore it was named camel milk bioactive peptide M2; the IC50 values ​​of components M1 and M3 were... 50 The values ​​were 4.36 mg / mL and 13.62 mg / mL, respectively.

[0050] (4) The M2 component was purified by preparing a 20 mg / mL aqueous solution and then subjected to preparative reversed-phase high-performance liquid chromatography (RP-HPLC). A C18 column was used. Mobile phase A was ultrapure water containing 0.2% trifluoroacetic acid, and mobile phase B was acetonitrile containing 0.1% trifluoroacetic acid. The mobile phases were: 0% mobile phase B, 0-6 min; 60-10% (6-30 min); 10-0% (30-35 min).

[0051] The flow rate was 0.5 mL / min, and the fraction was collected by detection at 280 nm combined with mass spectrometry. After six rounds of repeated purification, high-purity fraction K3 was finally obtained, with a peptide purity of 95.6 g / 100 g, significantly improved lipase inhibitory activity, and an IC50 concentration of [missing value]. 50 The value was 0.08 mg / mL. LC-ESI-Orbitrap MS / MS analysis revealed that the major peptide sequence in the K3 fraction was VQPIPY, with an abundance of 88.67% in the sample.

[0052] Example 2: Preparation of meal replacement powder containing active peptide M2 ​​(M2 component obtained in Example 1) 1. Preparation of meal replacement powder Meal replacement powder formula one: The meal replacement powder is composed of the following ingredients by weight percentage: 20% whey powder, 15% milk powder, 20% oats, 10% spinach powder, 10% allulose, 20% Job's tears powder, 1% white kidney bean extract, 3% inulin, and 1% active peptide M2.

[0053] The preparation method includes the following steps: 1) Raw material pretreatment: Whey powder, milk powder, oat powder, Job's tears powder, spinach powder, etc. are passed through an 80-100 mesh sieve to ensure uniform powder fineness.

[0054] 2) Weighing and mixing: Weigh each raw material accurately according to the above ratio, and use the equal increment method to gradually mix the trace components with the large amount of base powder evenly.

[0055] 3) Final mixing and sterilization: The pre-mixed materials are fed into a three-dimensional motion mixer and mixed for 30 minutes to ensure complete homogeneity. Sterilization is performed using cobalt-60 irradiation.

[0056] 4) Packaging: Packed into 25g bags in a cleanroom.

[0057] Meal replacement powder formula two: The preparation method for the meal replacement powder is the same as Formula 1, except that Formula 2 consists of the following ingredients by weight percentage: The formula contains 25% whey powder, 20% milk powder, 20% oats, 5% spinach powder, 5% allulose, 20% Job's tears powder, 2% white kidney bean extract, 1% inulin, and 2% active peptide M2.

[0058] Meal replacement powder formula three: The preparation method for the meal replacement powder is the same as that of Formula 1, except that Formula 3 consists of the following ingredients by weight percentage: The formula contains 25% whey powder, 15% milk powder, 15% oats, 10% spinach powder, 10% allulose, 20% Job's tears powder, 2% white kidney bean extract, 1% inulin, and 2% active peptide M2.

[0059] Comparative Example 1 The preparation method of the meal replacement powder is the same as that of Formula 1, except that it does not contain active peptides and is replaced by an equal amount of maltodextrin. The other ingredients and contents are exactly the same as those in Example 1.

[0060] Comparative Example 2 The preparation method of the meal replacement powder is the same as that of Formula 1, except that it does not contain allulose and white kidney bean extract, but is replaced with an equal amount of maltodextrin. The other ingredients and contents are exactly the same as those in Example 1.

[0061] 2. Effect Verification Experiment To verify the weight loss effect of the product of this invention, the following animal experiments were conducted in accordance with the research methods for the weight loss effects of traditional Chinese medicine compound prescriptions provided in the background art: Experimental methods Seventy healthy SPF-grade male C57BL / 6J mice were randomly divided into seven groups: blank control group, model control group, formula group 1, formula group 2, formula group 3, comparative group 1, and comparative group 2, with 10 mice in each group. Except for the blank control group, which was fed a normal diet, the other groups were fed a high-fat diet. Eight weeks after modeling, formula groups 1, 2, and 3, and comparative groups 1 and 2 were given the corresponding meal replacement powder (calculated at 10 times the recommended human dose, specifically 20 mg / gday), while the blank control group and model control group received an equal volume of physiological saline. This intervention continued for four weeks. Body weight was measured weekly, and the epididymal fat coefficient was determined at the end of the experiment.

[0062] Experimental results: Table 1: Changes in body weight of mice in each group (g, x±s)

[0063] Table 2: Comparison of epididymal fat coefficients in different groups of mice after the experiment (%, x±s)

[0064] Note: Compared with the model control group, *P<0.05, **P<0.01; compared with Example 1 group, #P<0.05, ##P<0.01.

[0065] The results show: (1) As can be seen from Table 1 and Table 2, the formula group 1 has the most significant effect in terms of weight loss and fat coefficient reduction, which is significantly better than each pair of ratio groups (P<0.01).

[0066] (2) Although the comparative group 1 (without active peptides) had a certain weight loss effect, it was significantly different from the formula group 1 (P<0.01), indicating that the active peptides played an important role in the weight loss process.

[0067] (3) The weight loss effect of the comparative group 2 (without allulose and white kidney bean extract) was also significantly lower than that of the formula group 1 (P<0.01), which proves the necessity of sugar-blocking and fat-controlling ingredients in the weight loss process.

[0068] Example 3: Application of the bioactive peptide VQPIPY in lowering blood lipids 1. Verification of lipid-lowering activity (in vivo mouse experiment) The animal model was established and grouped as follows. The high-fat diet formula involved added 20% lard, 1.2% cholesterol, and 0.2% sodium cholate to the basal diet; the specific steps are as follows: 1. Solution preparation Active peptide solution: The active peptide was dissolved in pure water to prepare VQPIPY active peptide aqueous solutions with concentrations of 5, 10, and 20 mg / ml.

[0069] 2. Animal experiments: Sixty healthy male ICR mice (weighing 18-22 g) were randomly divided into 6 groups (n=10), as follows: (A) Normal control group (fed basal diet); from week 1 to week 8, the basal diet was fed daily, and 0.2 ml of physiological saline was administered by gavage once. (B) High-fat model group (fed high-fat diet); from week 1 to week 8, the high-fat diet was fed daily, and 0.2 ml of physiological saline was administered by gavage once. (C) Positive control group (high-fat diet + simvastatin, 10 mg / kg / d); from week 1 to week 8, the diet was high-fat and 0.2 ml of simvastatin was administered by gavage once a day. The final gavage dose was 10 mg / kg / d. (D) Low-dose peptide group (high-fat diet + active peptide VQPIPY, 50 mg / kg / d); from week 1 to week 8, the diet was high-fat and 0.2 ml of active peptide VQPIPY solution was administered by gavage once a day. The final gavage concentration was 50 mg / kg / d. (E) Medium-dose peptide group (high-fat diet + active peptide VQPIPY, 100 mg / kg / d); from week 1 to week 8, the diet was high-fat and 0.2 ml of active peptide VQPIPY solution was administered by gavage once a day. The final gavage concentration was 100 mg / kg / d. (F) High-dose peptide group (high-fat diet + active peptide VQPIPY, 200 mg / kg / d). From week 1 to week 8, the diet was high-fat, and 0.2 ml of active peptide VQPIPY solution was administered by gavage once a day. The final gavage concentration was 200 mg / kg / d. 3. Sample collection and indicator detection: After the experiment, the mice were fasted for 12 hours, and blood was collected from their eyeballs. After standing, the blood was centrifuged to separate the serum. The levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in the serum were measured using a fully automated biochemical analyzer.

[0070] The results are shown in Table 3 below: Table 3: Experimental Results

[0071] Note: ^# indicates P < 0.01 compared to the normal control group; * indicates P < 0.05 compared to the high-fat model group. As shown in Table 3 above, the bioactive peptide VQPIPY obtained from the hydrolysis and purification of camel milk can effectively reverse the abnormal blood lipids induced by a high-fat diet in mice, and has a clear lipid-lowering effect.

[0072] Compared with the high-fat model group, all dose groups of the active peptide VQPIPY significantly reduced serum TC, TG, and LDL-C levels (P<0.05 or P<0.01) in a dose-dependent manner. The high-dose group (200 mg / kg) showed the most significant effect, reducing TC, TG, and LDL-C by approximately 31.5%, 34.2%, and 40.1%, respectively, while increasing HDL-C levels by approximately 25.8%. Its lipid-lowering effect was comparable to that of the positive control drug simvastatin, and no significant abnormal behavior or abnormal weight loss was observed in mice throughout the experiment, indicating good safety.

[0073] Example 4: Application of the bioactive peptide VQPIPY in liver protection and study of its antioxidant properties The specific steps are as follows: 1. Determination of the antioxidant activity of the bioactive peptide VQPIPY (1) DPPH free radical scavenging ability The free radical scavenging capacity of DPPH (1,1-diphenyl-2-trinitrohydrazino-DPPH) was determined using a standard method. 2 mL of sample solution (gradual concentration) and 2 mL of 0.2 mmol / L DPPH solution were added to a test tube, mixed thoroughly, and reacted in the dark for 30 min. The mixture was then centrifuged at 3000 r / min for 10 min, and the OD 517 nm value was measured and recorded as A. s The blank group, where anhydrous ethanol was used instead of DPPH solution, was denoted as A0, and the control group, where distilled water was used instead of sample solution, was used. The DPPH free radical scavenging rate (%) was calculated, with each group repeated three times.

[0074] The results show: DPPH radicals are very stable free radicals, and DPPH scavenging ability is usually expressed as IC50. The results showed that the IC50 values ​​of (tert-butylhydroquinone)TBHQ and the bioactive peptide VQPIPY were 95.67 and 83.62 μg / mL, respectively. The bioactive peptide VQPIPY exhibited strong DPPH radical scavenging activity. (2) Determination of hydroxyl radical scavenging ability Prepare 100 mL each of FeSO4 solution (6 mmol / L), H2O2 solution (6 mmol / L), and salicylic acid-ethanol solution (9 mmol / L); prepare 25 mL of peptide solution. The total reaction volume is 4 mL, divided into 3 groups: Sample group (A1): 1 mL of pigment solution, 1 mL of H2O2, 1 mL of FeSO4, and 1 mL of salicylic acid-ethanol solution were added sequentially; Blank group (A0): 1 mL of distilled water was used instead of the pigment solution, and other components were the same as above; Control group (A2): 1 mL of distilled water was used instead of H2O2, and other components were the same as the sample group. After mixing, each group was placed in a 40 ℃ water bath for 1 h, and the absorbance was measured at 251 nm. Each experiment was repeated 3 times. The hydroxyl radical scavenging rate was calculated.

[0075] The results show: Hydroxyl radicals are important initiators of liposome peroxidation. Hydroxyl radicals are highly oxidizing free radicals; the higher the scavenging capacity of hydroxyl radicals, the stronger the antioxidant capacity. At 400 μg / mL, the bioactive peptide VQPIPY exhibited stronger hydroxyl radical scavenging activity (72.61%) compared to TBHQ (69.81%).

[0076] (3) Determination of superoxide anion free radical scavenging ability To determine the superoxide anion radical scavenging capacity, 0.5 mL of sample solution and 1.5 mL of Tris-HCl solution were added to a test tube and mixed thoroughly in a 25°C water bath for 20 min. Then, 200 μL of 25 mmol / L pyrogallol was added, followed by 250 μL of HCl to stop the reaction. The mixture was then centrifuged at 3000 r / min for 10 min, and the OD was measured. 325 Measure and record as A 样本 The superoxide anion free radical scavenging rate was calculated, with each group repeated 3 times.

[0077] The results show: Studies have shown that peptide-separated fractions can effectively scavenge superoxide anion free radicals. When the concentration of the active peptide VQPIPY was 200 μg / mL, the scavenging rate of superoxide anions reached 81.30%. At a concentration of 200 μg / mL, the scavenging rate of TBHQ was 78.86%. The active peptide VQPIPY exhibits strong scavenging activity.

[0078] 2. Animal experiments on the hepatoprotective activity of the bioactive peptide VQPIPY (1) Grouping and feeding of mice Mice were housed in a laboratory animal center (SPF grade) at a temperature of 23±3℃ and a relative humidity of 55±15%, with a regular 12-hour light-dark cycle. Experiments and model establishment were conducted according to the methods outlined in the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)". After one week of acclimatization, the mice were randomly divided into six groups: blank control group (CON), model control group (MOD), positive control group (POS), low-dose group (FL), medium-dose group (FM), and high-dose group (FH).

[0079] The specific experiment is as follows: The first 7 days are the adaptation period. From day 8 to day 35, follow the instructions below: Blank control group (CON): CON group was given 0.1 ml of distilled water by gavage every day; Model control group (MOD): The MOD group was given 0.1 ml of distilled water by gavage daily; Positive control group (POS, biphenyl diester, 150 mg / kg BW): 0.1 mL of biphenyl diester aqueous solution was administered by gavage daily at a dose of 150 mg / kg BW; Low-dose group (FL): VQPIPY polypeptide aqueous solution was administered by gavage at a dose of 10 mL / kg BW daily, with the polypeptide dose being 5 mg / kg BW. Medium dose group (FM): VQPIPY polypeptide aqueous solution was administered by gavage at a dose of 10 mL / kg BW per day, with the polypeptide dose being 10 mg / kg BW. High-dose group (FH): VQPIPY polypeptide aqueous solution was administered by gavage at a dose of 10 mL / kg BW daily, with the polypeptide dose being 15 mg / kg BW. Six hours after the last administration, the CON group was treated by gavage with distilled water (gavage dose was the same as other groups), while the mice in the other groups were treated by gavage with 50% ethanol at a dose of 12 mL / kg BW.

[0080] (2) Determination of serum biochemical indicators and liver indicators After the animal experiments were completed (followed by gavage with alcohol or distilled water), the mice were fasted overnight and then anesthetized with a mixture of 1-1.5% ether and oxygen. Once fully anesthetized, blood was collected from the eyes. The mice were then euthanized by cervical resection. The blood sample was centrifuged at 3500 r / min for 15 min at 4°C to obtain mouse serum, and the levels of AST and ALT in the serum were measured. The liver tissue of the mice was removed and weighed to calculate the liver index. The calculation formula is as follows.

[0081] Liver index = liver mass / (mouse body weight) × 100%.

[0082] Experimental results 1) Effects of VQPIPY peptide on serum biochemical parameters in mice Serum biochemical markers ALT and AST levels were measured, and the results are shown in Table 4: Table 4: Experimental Results

[0083] The results show: Normally, ALT and AST are mainly found in the liver. However, when excessive alcohol is consumed, liver cell membranes are damaged, and ALT and AST are released into the bloodstream, leading to abnormally high levels of ALT and AST in the serum.

[0084] Compared with the CON group, the serum ALT and AST activities of mice in the alcohol MOD group were significantly increased (p<0.05), with ALT activity reaching 39.6 U / L and AST activity reaching 50.9 U / L. This indicates that the MOD model of acute alcoholic liver injury in mice has been successfully constructed.

[0085] Compared with the MOD group, the activities of ALT and AST in the FL, FM, and FH experimental groups were significantly decreased (p<0.05), indicating that VQPIPY peptides can effectively resist the damage of alcohol intake to hepatocytes, maintain the integrity and permeability of the cell membrane, and reduce the leakage of ALT and AST. It has a significant protective effect against alcoholic liver injury.

[0086] 2) Effects of peptides on liver indices in mice Compared with the CON group, the liver index of mice in the MOD group was significantly increased by 0.77% (p<0.05), preliminarily indicating that acute intragastric alcohol administration induced liver injury in mice, and the experimental model was successful. The liver index of the POS group and the peptide group was lower than that of the MOD group, indicating that the VQPIPY peptide alleviated acute alcoholic liver injury in mice.

[0087] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. The use of VQPIPY peptide or products containing VQPIPY peptide in the preparation of lipid-lowering drugs or health products, or in the preparation of drugs for treating and / or alleviating alcoholic liver injury, wherein the amino acid sequence of the VQPIPY peptide is shown in SEQ ID NO.

1.

2. The application according to claim 1, characterized in that, The product containing VQPIPY peptide is a meal replacement powder containing VQPIPY peptide. Preferably, the meal replacement powder formula is as follows: 20-25 parts whey powder, 15-20 parts milk powder, 15-25 parts oats, 5-15 parts spinach powder, 5-15 parts allulose, 15-25 parts coix seed powder, 0.5-2 parts white kidney bean extract, 1-5 parts inulin, and 1-2 parts VQPIPY peptide.

3. The application according to claim 1, characterized in that, The product containing VQPIPY peptide is a meal replacement powder containing VQPIPY peptide. Preferably, the meal replacement powder formula is as follows: 20-25 parts whey powder, 15-20 parts milk powder, 15-25 parts oats, 5-15 parts spinach powder, 5-15 parts allulose, 15-25 parts coix seed powder, 0.5-2 parts white kidney bean extract, 1-5 parts inulin, and 1-2 parts M2 component containing VQPIPY peptide; The preparation method of the M2 component containing VQPIPY peptide is as follows: (1) Add bromelain to camel milk for enzymatic hydrolysis. The substrate concentration is 1040 g / L, the enzyme addition amount is 500-10000 U / g, the hydrolysis temperature is 35-55 ℃, the hydrolysis time is 1-5 h, and the hydrolysis pH is 5.5-7.5 to obtain the hydrolysis product. (2) The enzymatic hydrolysis product was further separated by Sephadex G-15 dextran gel column: the column was pre-equilibrated with Tris-acetate buffer, and then pure water was used as the mobile phase. The ultraviolet absorption was monitored at a wavelength of 280 nm, and the eluted fraction was collected according to the absorption peak. When the elution time was 171 min, the M2 fraction containing VQPIPY peptide was obtained.

4. The application according to claim 1, characterized in that, The drug also contains pharmaceutically acceptable excipients; the excipients refer to conventional drug carriers in the pharmaceutical field. Preferably, the excipients include one or more of the following: binders such as cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, and mannitol; fillers such as starch and sucrose; humectants such as glycerin; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, fatty acid sorbitan, and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, and pharmaceutical iron oxide red; lubricants such as hydrogenated vegetable oil, talc, and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, and cellulose acetate; and other excipients such as flavoring agents and sweeteners may also be added to the composition.

5. The application according to claim 4, characterized in that, The dosage forms of the medicine include, but are not limited to, oral dosage forms, injectable dosage forms, and inhaled dosage forms; Preferably, the oral dosage form includes, but is not limited to, tablets, capsules, granules, oral liquids, and oral suspensions; Preferably, the injectable dosage form includes, but is not limited to, injection solution and injection powder for injection; Preferably, the inhalation dosage form includes, but is not limited to, aerosols and powder inhalers.

6. The application according to claim 5, characterized in that, The health products also contain acceptable excipients.

7. A drug or health product for lowering blood lipids, or a drug for treating and / or alleviating alcoholic liver damage, characterized in that, Its active ingredient is the bioactive peptide VQPIPY peptide.

8. The pharmaceutical product according to claim 7, characterized in that, The drug also contains pharmaceutically acceptable excipients; the excipients refer to conventional drug carriers in the pharmaceutical field. Preferably, the excipients include one or more of the following: binders such as cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, and mannitol; fillers such as starch and sucrose; humectants such as glycerin; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, fatty acid sorbitan, and fatty acid glycerides; colorants such as titanium dioxide, sunset yellow, methylene blue, and pharmaceutical iron oxide red; lubricants such as hydrogenated vegetable oil, talc, and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, and cellulose acetate; and other excipients such as flavoring agents and sweeteners may also be added to the composition.

9. The pharmaceutical product according to claim 8, characterized in that, The dosage forms of the medicine include, but are not limited to, oral dosage forms, injectable dosage forms, and inhaled dosage forms; Preferably, the oral dosage form includes, but is not limited to, tablets, capsules, granules, oral liquids, and oral suspensions; Preferably, the injectable dosage form includes, but is not limited to, injection solution and injection powder for injection; Preferably, the inhalation dosage form includes, but is not limited to, aerosols and powder inhalers.

10. The health product according to claim 7, characterized in that, The health products also contain acceptable excipients.

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