Anti-aging composition containing plant polypeptide and preparation method thereof

Abstract

The invention relates to an anti-aging composition containing plant polypeptide and a preparation method thereof. The tartary buckwheat polypeptide with the anti-oxidation characteristic is obtained from tartary buckwheat grains through enzymolysis and purification, the polypeptide has the good anti-oxidation characteristic and biological safety, the swimming score of a mouse can be remarkably improved, the activity of antioxidant enzyme in serum can be remarkably affected, and therefore the anti-aging effect is improved. The anti-aging pharmaceutical composition prepared from the polypeptide has a relatively wide application value.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and specifically to an anti-aging composition containing plant polypeptides and a preparation method thereof. Background Art

[0002] Anti-aging research is one of the core directions in the fields of biomedicine and cosmetics. In recent years, plant polypeptides have gradually become a hot topic in the development of anti-aging compositions due to their natural origin, high biological activity, and low toxicity. Plant polypeptides are usually extracted from plants such as soybeans, oats, and rosemary, and their anti-aging mechanisms cover multiple aspects such as antioxidant, promoting collagen production, and regulating cell metabolism. Combining the synergistic effects of natural plant extracts, such compositions show significant potential in delaying skin aging and improving body functions.

[0003] Plant polypeptides are widely present in plant seeds, leaves, stems, and processing by-products, and polypeptides from different plant sources have unique biological activities. Soybeans, peas, broad beans, etc. are important sources of plant polypeptides. For example, antibacterial peptides can be obtained by enzymatic hydrolysis of broad bean protein, with a molecular weight in the range of 1-3 kDa, which has an inhibitory effect on a variety of pathogenic bacteria. Soy peptides are rich in glutamine and arginine, and can promote skin barrier repair and collagen synthesis. Proteins in cereal bran such as oats, wheat, and corn can release antioxidant peptides after hydrolysis. Oat polypeptides are often used in the development of moisturizing anti-aging products due to their high hydrophilicity and low allergenicity. Polypeptides extracted from medicinal plants such as rosemary, green tea, and ginseng have both antioxidant and anti-inflammatory functions. Rosmarinic acid peptides delay the process of skin photoaging by scavenging free radicals and inhibiting the activity of matrix metalloproteinases (MMPs). In recent years, microalgae such as Spirulina and Chlorella have become emerging polypeptide sources due to their high protein content (60%-70%). For example, Spirulina peptides can enhance cell antioxidant capacity by activating the Nrf2 pathway. CN111544333B also discloses that the polypeptide composition consists of oat polypeptide and rice polypeptide, and the mass ratio of oat polypeptide to rice polypeptide is (5-6):(4-5). After compounding oat polypeptide and rice polypeptide according to a specific ratio, a significant synergistic effect is produced in terms of antioxidant efficacy.

[0004] The anti-aging mechanism of plant polypeptides mainly includes the following aspects: (1) Antioxidation and free radical scavenging. Plant polypeptides delay cellular oxidative damage by scavenging reactive oxygen species (ROS) and inhibiting lipid peroxidation. For example, the scavenging rate of rosemary extract on DPPH free radicals exceeds 90% at a concentration of 2 mg / mL. Polyphenolic plant components (such as resveratrol) enhance the expression of antioxidant enzymes by activating the Nrf2 pathway, forming a synergistic effect with polypeptides. (2) Promotion of collagen synthesis. Plant polypeptides directly promote the production of collagen and elastin by stimulating the proliferation of fibroblasts. For example, certain polypeptide compositions can increase the yield of collagen fibers by more than 30%. (3) Regulation of the cellular senescence signaling pathway. Polypeptides can delay cellular senescence by inhibiting the p16 / p53 pathway or activate autophagy-related genes (such as mTOR) to eliminate senescent cells. (4) Enhancement of the skin barrier function. Liposome encapsulation technology (such as cholesterol-phosphatidylcholine composite carriers) can improve the transdermal absorption rate of polypeptides and enhance their repair effect in the epidermal layer.

[0005] The preparation of plant polypeptides needs to balance the requirements of activity retention and large-scale production. Currently, the mainstream technologies include enzymatic hydrolysis, genetic engineering synthesis, and physical-assisted extraction. Enzymatic hydrolysis: This is the most commonly used method. Specific proteases (such as alkaline protease, trypsin) are selected to hydrolyze plant proteins to generate target polypeptides. For example, using a composite enzyme (alkaline protease + flavor protease) to hydrolyze soybean protein step by step can increase the polypeptide yield to more than 85% while reducing the production of bitter peptides. Optimizing conditions (such as temperature 45 - 55 °C, pH 8.0 - 9.0) can further improve the enzymatic hydrolysis efficiency. Genetic engineering synthesis: For polypeptides with specific functions (such as collagen-promoting peptides), they can be designed and synthesized through recombinant DNA technology. For example, using the Escherichia coli expression system to produce elastin-like polypeptides (ELPs) containing repetitive sequences, whose thermoresponsive properties can be used for the construction of intelligent delivery systems. Physical-assisted technologies: Physical means such as ultrasonic waves and high-pressure homogenization can disrupt plant cell walls and improve the protein release rate. Research shows that ultrasonic pretreatment (20 kHz, 300 W) can increase the enzymatic hydrolysis efficiency of wheat bran protein by 30%. In addition, supercritical CO 2 extraction technology is used for the separation of lipophilic polypeptides to avoid the problem of organic solvent residues. Purification and stabilization: Membrane filtration (ultrafiltration, nanofiltration) and chromatographic techniques (ion exchange, affinity chromatography) are key steps in polypeptide purification. For example, using a 10 kDa ultrafiltration membrane can separate anti-aging active peptide 6 with an appropriate molecular weight. To improve the stability of polypeptides, microencapsulation (such as liposomes, chitosan nanoparticles) and freeze-drying technology are widely used, which can extend the shelf life and enhance the transdermal absorption efficiency.

[0006] Although there have been certain achievements in the research on anti-aging polypeptides, multi-functional anti-aging compositions with high activity and no toxic side effects still need to be further developed and studied. Summary of the Invention

[0007] The present invention overcomes the deficiencies of the prior art and provides an anti-aging composition containing plant polypeptide and a preparation method thereof.

[0008] Specifically, the plant polypeptide is tartary buckwheat polypeptide hydrolyzed from tartary buckwheat seed protein and has antioxidant biological activity.

[0009] More specifically, the plant polypeptide of the present invention is tartary buckwheat polypeptide TB-3-B, and its amino acid sequence is as shown in SEQ ID NO: 1.

[0010] Specifically, the present invention also provides an anti-aging composition, and the composition contains tartary buckwheat polypeptide TB-3-B, and its amino acid sequence is as shown in SEQ ID NO: 1.

[0011] Furthermore, the present invention also provides the use of tartary buckwheat polypeptide TB-3-B in the preparation of an anti-aging composition, wherein the tartary buckwheat polypeptide TB-3-B has an amino acid sequence as shown in SEQ ID NO: 1.

[0012] More specifically, the composition further contains a pharmaceutically acceptable carrier.

[0013] The pharmaceutical composition of the present invention or the pharmaceutical composition in the drug package can be made into various dosage forms suitable for administration routes such as oral, inhalation, rectal, topical, parenteral, etc., including but not limited to: powders, tablets (including various coated tablets, sustained-release or controlled-release tablets), lozenges, capsules (including soft capsules and hard capsules), granules, pills, dispersible powders, aqueous or oily suspensions, aqueous or oily solutions, emulsions, elixirs, syrups, etc. suitable for oral administration; powder or liquid aerosols suitable for inhalation; suppositories suitable for rectal administration; creams, ointments, gels, aqueous or oily solutions, aqueous or oily suspensions, etc. suitable for topical administration; sterile aqueous or oily injections or lyophilized powder injections for intravenous, subcutaneous or intramuscular injection suitable for parenteral administration. The present invention can also administer the independent drugs in the form of a drug package, administer the independent components in different dosage forms or at different dosing intervals.

[0014] Pharmaceutically acceptable carriers include, but are not limited to, fillers (or diluents), binders, disintegrants, lubricants, wetting agents, auxiliary lipids, glidants, sweeteners, flavoring agents, solvents, solubilizing aids, suspending agents, isotonic agents, buffers, preservatives, antioxidants, coloring agents, foaming agents, etc. Those skilled in the art can select the above-mentioned pharmaceutically acceptable carriers according to actual needs. For example, fillers (diluents) that can be used include, but are not limited to, lactose, sucrose, microcrystalline cellulose, starch, mannitol, mannitol-starch, etc.; binders that can be used include, but are not limited to, microcrystalline cellulose, polyvinylpyrrolidone, tragacanth gum, glucose solution, gum arabic paste, gelatin solution, sucrose, starch paste, etc.; disintegrants that can be used include, but are not limited to, croscarmellose sodium, low-substituted hydroxypropyl cellulose, crospovidone, sodium starch glycolate, alginic acid, dry starch, bentonite, methyl cellulose, agar, carboxymethyl cellulose, etc.; lubricants that can be used include, but are not limited to, talc, magnesium or calcium stearate, lycopodium, etc.; wetting agents that can be used include, but are not limited to, propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether, etc.; auxiliary lipids that can be used include, but are not limited to, phosphatidylethanolamine, phosphatidylcholine, cholesterol, etc.; glidants that can be used include, but are not limited to, colloidal silica, etc.; sweeteners that can be used include, but are not limited to, sucrose, lactose mannitol, artificial sweeteners (such as sodium cyclamate and saccharin), etc.; flavoring agents that can be used include, but are not limited to, mint and methyl salicylate, etc.

[0015] More specifically, the composition further contains other antioxidant substances.

[0016] Examples of the other antioxidant substances include, but are not limited to, hydrolyzed collagen, hydrolyzed elastin, yeast extract, oryzanol, tetrahydrocurcumin, ellagic acid, whey protein, salicyl phytosphingosine, silymarin, sericin, sodium tocopheryl phosphate, allantoin, ribonucleic acid (RNA), grape (VITIS VINIFERA) seed extract, ormosia henryi bark extract, tea polyphenols, wine extract, apple seed extract, fagus sylvatica bud extract, hydrolyzed baobab extract, artemia (ARTEMIA) extract, iris germanica root extract, hesperidin, ginsenoside, salvia miltiorrhiza extract, niacinamide, ursolic acid, lycopene, coffee extract, lactic acid, superoxide dismutase (SOD), evening primrose oil, ceramide, dipalmitoyl hydroxyproline, hydroxystearic acid, salicylic acid, ergothioneine, lysophosphatidylcholine, lipoic acid, glycogen, resveratrol, ferulic acid, lysate of bifida ferment lysate, lysate of lactic acid bacteria fermentation, etc. The content of the other antioxidant substances in the composition is known in the art. For example, it usually accounts for 0.01-10% of the total weight of component (C).

[0017] More specifically, the pharmaceutically acceptable carrier includes at least one of a stabilizer, a suspending agent, a pH regulator, an osmotic pressure regulator, and a lyoprotectant.

[0018] The concentration range of the stabilizer can be 5.0 mg / mL - 48.0 mg / mL. In some embodiments, the concentration of the stabilizer is 10.0 mg / mL - 35.0 mg / mL. In some examples, the concentration range of the stabilizer is 6.0 mg / mL - 10.0 mg / mL; in some examples, the concentration range of the stabilizer is 6.0 mg / mL - 15.0 mg / mL; in some examples, the concentration range of the stabilizer is 6.0 mg / mL - 20.0 mg / mL; in some examples, the concentration range of the stabilizer is 6.0 mg / mL - 30.0 mg / mL; in some examples, the concentration range of the stabilizer is 6.0 mg / mL - 35.0 mg / mL; in some examples, the concentration range of the stabilizer is 10.0 mg / mL - 15.0 mg / mL; in some examples, the concentration range of the stabilizer is 10.0 mg / mL - 20.0 mg / mL; in some examples, the concentration range of the stabilizer is 10.0 mg / mL - 30.0 mg / mL; in some examples, the concentration range of the stabilizer is 10.0 mg / mL - 35.0 mg / mL; in some examples, the concentration range of the stabilizer is 10.0 mg / mL - 48.0 mg / mL; in some examples, the concentration range of the stabilizer is 15.0 mg / mL - 20.0 mg / mL; in some examples, the concentration range of the stabilizer is 15.0 mg / mL - 30.0 mg / mL; in some examples, the concentration range of the stabilizer is 15.0 mg / mL - 35.0 mg / mL; in some examples, the concentration range of the stabilizer is 15.0 mg / mL - 48.0 mg / mL; in some examples, the concentration range of the stabilizer is 20.0 mg / mL - 30.0 mg / mL; in some examples, the concentration range of the stabilizer is 20.0 mg / mL - 35.0 mg / mL; in some examples, the concentration range of the stabilizer is 20.0 mg / mL - 48.0 mg / mL; in some examples, the concentration range of the stabilizer is 30.0 mg / mL - 35.0 mg / mL; in some examples, the concentration range of the stabilizer is 30.0 mg / mL - 48.0 mg / mL; in some examples, the concentration range of the stabilizer is 35.0 mg / mL - 48.0 mg / mL; in some examples, the concentration range of the stabilizer is 5.0 mg / mL - 25.0 mg / mL. In some examples, the concentration of the stabilizer is 7.9 mg / mL, 14.5 mg / mL, 15.0 mg / mL, 17.0 mg / mL, 20.0 mg / mL, 30.0 mg / mL, or 35.0 mg / mL.

[0019] The carrier further includes a suspending agent. The suspending agent includes at least one selected from dextran, gelatin, hypromellose, methylcellulose, gum arabic, polyethylene glycol 3350, polyethylene glycol 4000, polyethylene glycol 6000, sodium carboxymethyl cellulose, and polyvinylpyrrolidone. In some embodiments, the suspending agent is polyethylene glycol 4000; in some embodiments, the suspending agent is polyethylene glycol 3350; in some embodiments, the suspending agent is polyethylene glycol 6000; in some embodiments, the suspending agent is sodium carboxymethyl cellulose; in some embodiments, the suspending agent is polyvinylpyrrolidone K12; in some embodiments, the suspending agent is polyvinylpyrrolidone K30.

[0020] The carrier also includes an acid-base regulator. In one embodiment, the acid-base regulator is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, or a combination thereof. In one embodiment, the acid-base regulator is a hydrochloric acid solution or a sodium hydroxide solution, such as a 1M hydrochloric acid solution or a 1M sodium hydroxide solution.

[0021] The pharmaceutical composition further contains a lyoprotectant; wherein, the lyoprotectant includes at least one selected from lactose, mannitol, glycine, sucrose, trehalose, maltose, xylitol, fructose, galactose, polyvinylpyrrolidone, polyethylene glycol, dextran, albumin, L-serine, sodium glutamate, alanine, sarcosine, arginine, and histidine. According to some embodiments of the present invention, the lyoprotectant is preferably lactose. Thus, by using the lyoprotectant of the embodiments of the present invention, the drug activity of the suspension containing ICS and LABA can be protected during the freeze-drying process, and the stability of the freeze-dried powder during storage at room temperature can be further improved, and the suspension after reconstitution of the freeze-dried powder can still be atomized.

[0022] Advantageous Effects

[0023] In the present invention, tartary buckwheat polypeptide with antioxidant properties is obtained by enzymatic hydrolysis and purification from tartary buckwheat grains. The polypeptide has good antioxidant properties and biological safety, can significantly improve the swimming score of mice, and significantly affect the activity of antioxidant enzymes in serum, thereby improving the anti-aging effect. Preparing the polypeptide into an anti-aging pharmaceutical composition has relatively broad application value. Brief Description of the Drawings

[0024] Figure 1 Graph showing the effect of the polypeptide of the present invention on cell viability

[0025] Figure 2 Graph showing the effect of the polypeptide of the present invention on the swimming test score of mice Detailed Description of the Embodiments

[0026] The present invention can be further described by the following examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art can understand that various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention. The present invention generally and / or specifically describes the materials and test methods used in the experiments. Although many materials and operation methods used to achieve the purpose of the present invention are well known in the art, the present invention still describes them in as much detail as possible herein. The following examples further illustrate the present invention rather than limit it. Any formal but non-substantive equivalent transformation made based on the concept of the present invention should be regarded as falling within the scope of the technical solution of the present invention.

[0027] Example 1 Separation and Identification of Tartary Buckwheat Polypeptides

[0028] Using fresh tartary buckwheat grains as raw materials, 1 kg of tartary buckwheat grains were soaked in 10 L of citric acid aqueous solution (citric acid concentration: 0.025 mol / L) for 20 h, and then heated in pure water at 95 °C for 100 min. The treated tartary buckwheat grains were ground into a paste and placed in an enzymatic hydrolysis tank, and pure water, trypsin, and alkaline protease were added. The addition amounts of pure water, trypsin, and alkaline protease were 6 times the mass of the tartary buckwheat grains, 0.6%, and 0.4% respectively. After stirring and mixing evenly, the pH value of the solution was adjusted to 8.0, and enzymatic hydrolysis was carried out at 55 °C for 4 h. Then, papain was added to the enzymatic hydrolysate, and the amount of papain was 0.4% of the mass of the tartary buckwheat grains. The pH value was adjusted to 7.0, and enzymatic hydrolysis was continued for 3 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 100 °C for 10 min and then quickly cooled. The obtained enzymatic hydrolysate was filtered through a 50-nm ceramic membrane and the permeate was collected. Then, reverse osmosis membrane filtration was used to collect the concentrate, and the tartary buckwheat enzymatic hydrolysate powder was obtained by freeze-drying.

[0029] The tartary buckwheat enzymatic hydrolysate powder was formulated into an 8 mg / mL solution and filtered through a 0.45-μm filter membrane. Then, it was separated by a Sephadex G-15 (1.6 cm × 80 cm) gel column. The sample loading volume was 4 mL, the flow rate was set at 0.5 mL / min, and 1 tube was collected every 4 min. Ultra-pure water was used for elution, and the absorbance value was detected at a wavelength of 214 nm with a UV spectrophotometer. The eluate components with similar absorbance values were collected and combined. A total of 7 components with similar wave peaks were obtained, named TB-1, TB-2, TB-3, TB-4, TB-5, TB-6, and TB-7 respectively, and each component was freeze-dried. The antioxidant effects of the above 7 components were tested by the ABTS free radical scavenging ability at a concentration of 100 μg / mL. The results are shown in Table 1.

[0030] Table 1 Detection Results of the Antioxidant Capabilities of Each Peak

[0031] Group Clearance rate (%) TB-1 34.52±2.54 TB-2 47.13±3.06 TB-3 78.63±5.17 TB-4 50.43±4.31 TB-5 34.78±2.54 TB-6 20.27±1.59 TB-7 10.23±0.23

[0032] As can be seen from the results in Table 1, the TB-3 component has the strongest antioxidant ability, and this component was selected for further separation.

[0033] RP-HPLC was used to separate and purify the TB-3 component with good activity. An Agilent 1100 Series high performance liquid chromatograph was used, and an EXL-126-2546U ACE Excel 5AQ (4.6×250 mm) chromatographic column was selected. The mobile phase was acetonitrile and distilled water, and the elution conditions were as follows: the proportion of the organic phase (acetonitrile) increased from 0% to 35% within 0-25 min, the flow rate was 1 mL / min, the absorbance value at a wavelength of 280 nm was detected, and three peaks, TB-3-A, TB-3-B, and TB-3-C, were collected. The ABTS free radical scavenging ability test was further carried out on these three peaks for antioxidant effect, and the results showed that the TB-3-B component had the strongest antioxidant ability. The polypeptide sequence of the TB-3-B component was identified by LC-MS / MS, and its amino acid sequence is shown as SEQ ID NO: 1. The polypeptide was further entrusted to Peptide Valley for artificial synthesis and adjusted to a concentration of 5 mg / mL for standby.

[0034] Example 2 Identification of the antioxidant properties of the TB-3-B polypeptide

[0035] RAW 264.7 cells, purchased from Punosai Biotechnology. The culture medium for RAW264.7 macrophages was RPMI-1640 containing 10% (w / w) fetal bovine serum and 1% (w / w) double antibody, and the culture conditions were: 37 °C, 5% CO 2 and 95% air humidity. The culture medium was changed every 2 days and passaged every 3 days. Before the experiment, the cell morphology was observed with an inverted microscope, and cells in the logarithmic growth phase were used for the experiment, with 6 parallels for each experiment.

[0036] Cells were seeded in a 96-well cell culture plate at a density of 1.5×10 5 cells / mL. After culturing for 24 h, they were randomly divided into a blank control group (normal cells), a hydrogen peroxide damage group (cells + 200 μmol / L H 2 O 2 ), and sample determination groups a-d (cells + samples with mass concentrations of 50 (a), 100 (b), 200 (c), and 500 μg / mL (d) + 200 μmol / L H 2 O 2 ), and a positive control group (cells + Vc with a mass concentration of 200 μg / mL + 200 μmol / L H 2 O 2)。The sample addition group first added 100 μL of samples with different mass concentrations and cultured for 24 h. After rinsing 3 times with D-hanks buffer solution, all were added to serum-free culture medium, and then 200 μmol / L of H 2 O 2 was added. After continuing to culture for 60 min, the cell survival rate was detected by the MTT method, and the results were as Figure 1 shown.

[0037] As Figure 1 shown, there was a significant difference between the injury control group and the blank control group (P<0.05), indicating that H 2 O 2 successfully induced oxidative damage in RAW264.7 macrophages. As the polypeptide mass concentration increased, the cell survival rate gradually increased. Under the action of 200 μg / mL polypeptide, the cell survival rate was (98.32±1.95)%. The polypeptide also had slightly higher antioxidant properties compared with the positive control group at the same concentration, which fully indicated that the polypeptide of the present invention had an obvious protective effect on H 2 O 2 induced oxidative damage in RAW264.7 macrophages.

[0038] Example 3 Animal experiment of TB-3-B polypeptide

[0039] Kunming mice, SPF grade, body weight 20-25 g, male, provided by Vital River. The animals were grouped and raised in a mouse house with 12 h light-dark alternation. The temperature of the mouse house was 18-22 °C, and the relative humidity was (50±5)%. They were fed with standard feed and could freely eat and drink. Grouping treatment: 40 male mice were randomly divided into 4 groups, with 10 mice in each group, namely the blank group (NC), the oxidative damage model group (FC), the TB-3-B polypeptide group (TB-3-B), and the positive control group (Vc). The TB-3-B group and the positive control group were respectively gavaged at 5 mL / (kg·d) (concentration of 1 mg / mL), once a day for a total of 28 d. The NC group and the FC group drank water and ate normally. The oxidative damage model group (FC), the TB-3-B polypeptide group (TB-3-B), and the positive control group (Vc) were all subcutaneously injected with D-galactose at 400 mg / kg every day for 28 d.

[0040] Anti-fatigue detection: Swimming experiment method: 30 min after gavage on the 28th day, the mice were put into a square swimming tank with a water depth of about 15 cm and a water temperature of about 25 °C, and allowed to swim freely. If the mice could swim continuously within 1 min, 300 points were recorded; if they swam continuously for more than 30 s and only floated occasionally, 2.5 points were recorded; if they floated for more than 30 s, 2.0 points were recorded; if they only swam occasionally, 1.5 points were recorded; if they floated continuously, 1.0 points were recorded. The average value of 5 experimental results was taken as the final score of the mice. The results were as Figure 2 shown.

[0041] The swimming experiment examined the exercise endurance of mice. If the mice could swim continuously in water, the higher the score, the stronger the exercise endurance and the more significant the anti-fatigue effect. From Figure 2 It can be seen that the swimming score of the oxidative damage model group was significantly lower than that of the blank group, while the swimming score of the group treated with TB-3-B polypeptide reached (2.92±0.21) points, which was higher than that of the positive control group (2.50±0.11) points, indicating that the polypeptide could improve the exercise endurance of mice.

[0042] After intragastric administration on the 28th day, blood was collected from the orbital cavity, centrifuged at 5000 r / min for 10 min, the serum was separated, and the activities of serum SOD, GSH-Px and the content of MDA were detected. The content of MDA was determined by the thiobarbituric acid (TBA) colorimetric method; SOD was determined by the xanthine oxidase method; the activity of GSH-Px was determined by the DTNB method. The results are shown in Table 2.

[0043] Table 2 Effects of MDA, SOD and GSH-Px in the serum of mice in each group

[0044]

[0045] As can be seen from Table 2, compared with the control group and each treatment group, the content of MDA in the serum of mice in the model group increased extremely significantly, and the activities of SOD and GSH-Px decreased significantly (P<0.05). Compared with the model group, the content of MDA in the serum of mice in the TB-3-B polypeptide group decreased significantly, and the activities of GSH-Px and SOD increased significantly. This also fully indicates that the TB-3-B polypeptide of the present invention can effectively improve the activity of endogenous antioxidant enzymes in the body, scavenge free radicals and inhibit peroxidative damage as an antioxidant drug.

[0046] Example 4 Safety detection of polypeptide

[0047] Acute toxicity test, maximum single-dose method. Ten healthy Kunming mice, 5 males and 5 females, were selected. The mice were fasted overnight for 16 h before the test without water restriction. After intragastric administration at a dose of 1 g / kg / d, they were continuously observed for 1 week, and the toxic manifestations and death conditions of the animals were recorded. The results showed that no obvious toxic symptoms were observed in the 10 mice and no death occurred, indicating that the polypeptide is a non-toxic substance and has good safety.

[0048] Specific embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although specific embodiments of the present invention are shown in the drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided so that the present invention can be more thoroughly understood and the scope of the present invention can be fully conveyed to those skilled in the art.

[0049] Although specific terms are used herein, they are used for general and descriptive purposes only and not for purposes of limitation. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter described in this disclosure belongs.

Claims

1. A tartary buckwheat polypeptide TB-3-B with antioxidant and anti-aging functions, characterized in that The amino acid sequence is shown in SEQ ID NO:

1.

2. An anti-aging composition, characterized in that Contains the tartary buckwheat polypeptide TB-3-B with anti-oxidation and anti-aging functions as claimed in claim 1.

3. Use of the tartary buckwheat polypeptide TB-3-B with antioxidant and anti-aging functions according to claim 1 in the preparation of an anti-aging composition.

4. The use according to claim 3, wherein Relative to the total weight of the anti-aging composition, the content of the polypeptide is 0.001 wt % to 50 wt %.

5. The use according to claim 3, wherein The anti-aging composition is a pharmaceutical composition or a cosmetic composition.

6. The use according to claim 5, wherein The pharmaceutical composition is formulated into an emulsion, an ointment, a gel, a cream, a patch or a spray.

7. The use according to claim 5, wherein The cosmetic composition is formulated into a softening lotion, an astringent lotion, a nourishing lotion, an eye cream, a nourishing cream, a massage cream, a cleansing cream, a cleansing foam, a cleansing water, a powder, an essence or a mask.

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