A composition and its application in inhibiting non-enzymatic glycosylation reaction

Abstract

The present invention provides a composition and its application in inhibiting non-enzymatic glycosylation reaction. The composition provided by the present invention comprises sialic acid and Poria cocos extract, which are active ingredients and can inhibit non-enzymatic glycosylation reaction and the formation and accumulation of advanced glycosylation end products AGEs, thereby preventing skin dullness and darkening and maintaining skin transparency. The combination can be applied to the preparation of anti-glycation compositions, and can also be used as an additive component for products such as medicines, cosmetics, and health foods.

Description

Technical Field

[0001] The present invention relates to the biotechnology field of inhibiting protein glycation in tissues, in particular to a composition and application thereof in inhibiting non-enzymatic glycation reactions. Background Art

[0002] Advanced glycation end products (AGEs) are stable covalent adducts formed when macromolecules such as proteins, lipids, or nucleic acids spontaneously react with glucose or other reducing monosaccharides without the involvement of enzymes. They are the end products of non-enzymatic glycosylation reactions (Maillard reactions) and are the product of excess sugars and proteins.

[0003] AGEs can cause pigmentation. Skin pigmentation is a phenomenon caused by damage to the skin barrier due to both internal stressors (e.g., changes in hormone levels) and external stressors (e.g., ultraviolet radiation). It, along with wrinkles, is considered a manifestation of skin photoaging. UV exposure to the skin induces oxidative stress, increasing the release of proinflammatory cytokines in the skin and ultimately producing AGEs. Crisan et al. demonstrated the distribution of AGEs in dermal fibroblasts and endothelial cells using immunohistochemistry. AGEs can form pigment adducts with proteins, leading to skin pigmentation.

[0004] AGEs can also contribute to skin aging. Human skin tissue contains abundant collagen and elastin, which primarily exist in a fibrous form and form the skin's skeletal structure, giving it a certain strength and elasticity. AGEs can form cross-links with these two proteins, impairing not only their normal function but also the adhesion and growth of skin cells. Furthermore, the accumulation of cross-links between AGEs and these two proteins in the cellular matrix reduces the permeability of connective tissue and impairs the diffusion of nutrients and metabolic waste within the body. This increases the firmness of skin tissue, resulting in a decrease in skin elasticity and ultimately leading to skin aging and wrinkles.

[0005] In addition, AGEs contribute to the development of many chronic degenerative diseases, such as diabetes, atherosclerosis, chronic kidney disease, liver disease, and neurodegenerative diseases. Reducing AGEs can help fight aging and prevent various chronic degenerative diseases.

[0006] Although some of the anti-glycation agents disclosed in the prior art, such as aminoguanidine, have been proven to be glycation inhibitors, aminoguanidine belongs to the pharmaceutical range and has adverse reactions. Other anti-glycation ingredients, such as vitamin C and tea polyphenols, are easily oxidized, which limits their anti-glycation effects and application forms. Therefore, the development or discovery of better anti-glycation active ingredients is a problem that needs to be solved at present. Summary of the Invention

[0007] The purpose of the present invention is to address the above-mentioned deficiencies in the prior art and to provide a composition containing sialic acid and Poria cocos extract and its application.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] The invention provides a composition comprising sialic acid and Poria cocos extract as active ingredients, wherein the weight percentage of sialic acid is 0.1-50%.

[0010] Furthermore, the mass ratio of the sialic acid to the Poria cocos extract is no greater than 1.

[0011] The composition of the present invention may further include carnosine, and the mass ratio of the sialic acid to the carnosine is not greater than 10.

[0012] Furthermore, the sialic acid includes N-acetylneuraminic acid and / or its salt.

[0013] Furthermore, the Poria extract includes Poria polysaccharide and its derivatives.

[0014] Furthermore, the pachymaran includes carboxymethyl pachymaran.

[0015] In one application case, the pachymaran is carboxymethyl pachymaran.

[0016] Furthermore, the degree of substitution of the carboxymethyl pachymaran is 0.2 to 0.9.

[0017] Furthermore, the composition is used in the preparation of advanced glycation end products (AGEs) inhibitors.

[0018] Furthermore, the addition amount of the composition in the preparation of advanced glycation end products (AGEs) inhibitor is 10 to 100 wt%.

[0019] The present invention also provides a cosmetic comprising the above composition.

[0020] The beneficial effects brought about by the technical solution provided by the present invention are:

[0021] (1) The composition provided by the present invention contains sialic acid and Poria extract. First, sialic acid has free amino groups that can compete with carbohydrates for proteins, acetylate the amino end, and reduce the amount of AGEs. Second, the free amino groups in sialic acid can react with the reducing carbonyl groups in the Amadori product to form inactive substances, eliminate the carbonyl groups, and thus prevent the Amadori product from further rearrangement. Therefore, sialic acid inhibits the formation and accumulation of advanced glycation end products AGEs, thereby inhibiting non-enzymatic glycosylation reactions. When sialic acid is compounded with Poria extract, the antioxidant effect of Poria extract inhibits the formation of early glycated protein products, and synergistically with sialic acid to prevent or delay the non-enzymatic glycosylation reaction of proteins and the formation of glycation end products, which can effectively inhibit the formation and accumulation of advanced glycation end products AGEs, inhibit non-enzymatic glycosylation reactions, help prevent skin dullness and darkening, and maintain skin transparency. It can be used to prepare anti-glycation products.

[0022] (2) The composition containing sialic acid and Poria cocos extract provided by the present invention can also improve product stability by inhibiting the glycosylation reaction (Maillard reaction) of amino acids and proteins contained in the product. By inhibiting the glycosylation reaction, the color tone of the product can be stabilized. It can be used as an additive in products such as medicines, cosmetics, and health foods. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a diagram showing the effect of the non-enzymatic glycosylation inhibition test of different concentrations of sialic acid provided by the present invention;

[0024] Figure 2 This is a graph showing the effect of a non-enzymatic glycosylation inhibition test of sialic acid compounded with Poria cocos extracts of different concentrations provided by the present invention;

[0025] Figure 3 This is a graph showing the effect of a non-enzymatic glycosylation inhibition test of a compound of sialic acid and Poria cocos extracts with different degrees of substitution provided by the present invention;

[0026] Figure 4 This is a diagram showing the effect of a non-enzymatic glycosylation inhibition test of a combination of sialic acid, Poria cocos extract and carnosine provided by the present invention. DETAILED DESCRIPTION

[0027] In order to make the objectives, technical solutions and advantages of the present invention more clear, the embodiments of the present invention will be further described below with reference to the accompanying drawings and examples.

[0028] The invention provides a composition capable of inhibiting non-enzymatic glycosylation reaction, comprising sialic acid and Poria cocos extract as active ingredients, wherein the weight percentage of sialic acid is 0.1-50%.

[0029] In order to fully utilize the efficacy of the composition containing sialic acid and Poria cocos extract and obtain a high saccharification effect, the mass ratio of sialic acid as an active ingredient to Poria cocos extract is not greater than 1.

[0030] That is, the mass ratio of sialic acid: Poria extract can be 1:1, 1:2, 1:4, 1:8, 1:16. The upper limit ratio can be determined according to actual usage, such as dissolution state, recommended intake, production cost and other factors, and is not limited here.

[0031] Sialic acid, used in the present invention, is a naturally occurring, biologically active nine-carbon sugar molecule, encompassing over 80 different species. It contains a pyranose structure and is a type of acidic amino sugar. It is widely present in animal tissues, typically in a mucus-bound form on the upper surface of human eukaryotic cells. It binds to glycoproteins and lipopolysaccharides in the outermost layer of the cell membrane, forming a tight protective barrier on the cell surface. It possesses potential anti-glycation activity. Research by the applicant has shown that sialic acid can effectively inhibit non-enzymatic glycosylation reactions.

[0032] The Poria cocos used in this invention is a member of the Polyporaceae family and lives at the roots of pine trees. Poria cocos is a fungus approximately 30-50 cm in diameter, with a rough exterior similar to pine bark and a white or pink interior. Poria cocos has a sweet, mild flavor and a neutral nature. It has been recognized as a top-quality beauty product, known for its benefits of strengthening the spleen and lungs, dispelling dampness, removing freckles, whitening, and moisturizing the skin. Its triterpenes have been reported to inhibit tyrosinase activity, thereby inhibiting melanin production in the body. Poria cocos sclerotia contain β-poria polysaccharide, equivalent to 93% of the dry weight of the Poria cocos. Poria cocos also contains triterpenoids, pachymic acid, pyrimethicone, and pinene acid, as well as keratin, sterols, lecithin, adenine, histidine, choline, lipase, and protease. Furthermore, modern pharmacological research has shown that the triterpenoids and polysaccharides in Poria cocos, as natural active substances, possess a variety of physiological activities, such as enhancing immunity, antibacterial, hypoglycemic, and anti-tumor effects.

[0033] In the present invention, research has found that one of the sialic acids, N-acetylneuraminic acid, exhibits a high anti-glycation effect and can effectively inhibit the formation and accumulation of advanced glycation end products (AGEs). This is because N-acetylneuraminic acid has special structural characteristics: a carboxyl group at position 1 and an amino group at position 5. The carboxyl group at position 1 competes with carbohydrates for the free amino group of proteins, acetylating the amino end and reducing the amount of AGEs; the amino group at position 5 undergoes an addition reaction with the reducing carbonyl group in the Amadori product to form an inactive substance, eliminating the carbonyl group, thereby preventing further rearrangement of the Amadori product; and the α-carbon on the amide group at position 5 can also undergo an aldol condensation reaction with an active α-dicarbonyl intermediate to form an aldol compound, thereby blocking the conversion of the Amadori product into AGEs. In the present invention, the source of N-acetylneuraminic acid or N-acetylneuraminic acid salt is not limited. The production method of N-acetylneuraminic acid includes synthesis (chemical synthesis, enzymatic synthesis), natural extraction (from raw materials, egg white, submandibular gland mucin, placental extract, fermentation; wherein the fermentation method can produce sialic acid monomer or polysialic acid, and its source does not affect the use. In this embodiment, N-acetylneuraminic acid is separated and purified from a material containing polysialic acid. The preparation process includes: using a material containing polysialic acid as a raw material, through sterilization and solids, protein removal, decolorization, hydrolysis, desalting, crystallization and other steps, a high-purity N-acetylneuraminic acid product can be obtained, the purity of which is not less than 98%, which can meet the requirements in the fields of food, health care, medicine and cosmetics. For details, reference may be made to the preparation method of N-acetylneuraminic acid of Chinese patent CN201811364040.9. N-acetylneuraminic acid salt can be prepared as sodium salt, magnesium salt, potassium salt, amino acid salt or amine salt according to production needs.

[0034] The study also found that when sialic acid and Poria extract are used as a composition, the effect of inhibiting the formation and accumulation of advanced glycation end products AGEs is significantly improved compared to the effect of single sialic acid. In the present invention, the source of Poria extract is not limited. Poria extract includes Poria polysaccharide and its derivatives. Preferably, Poria polysaccharide can be carboxymethyl Poria polysaccharide, and more preferably carboxymethyl Poria polysaccharide with a degree of substitution of 0.2 to 0.9. In the present embodiment, Poria extract can be prepared by separation and extraction from dry Poria block or Poria fermentation liquid. The preparation process includes: using dry Poria block or Poria fermentation liquid as raw material, and obtaining Poria extract by steps of alcohol extraction, filtration, concentration, drying, etc. The obtained Poria extract is subjected to deproteinization and column chromatography treatment to obtain high-purity Poria polysaccharide, which has good antioxidant and antibacterial capabilities. In order to better apply Poria cocos polysaccharide, the hydroxyl groups on the Poria cocos polysaccharide structure can be replaced with carboxymethyl groups. Carboxymethylated Poria cocos polysaccharide is carboxymethyl Poria cocos polysaccharide, which has good solubility and can better utilize its antioxidant and antibacterial biological activities. The applicant's research found that in a composition containing sialic acid and Poria cocos extract, carboxymethyl Poria cocos polysaccharide of different concentrations can significantly inhibit non-enzymatic glycation reactions, and the inhibitory effect increases with increasing concentration; carboxymethyl Poria cocos polysaccharide of different substitution degrees can significantly inhibit non-enzymatic glycation reactions, and the inhibitory effect increases with increasing substitution degrees, indicating that carboxymethyl Poria cocos polysaccharide is an effective AGEs inhibitor, mainly because the high antioxidant activity of carboxymethyl Poria cocos polysaccharide can inhibit non-enzymatic glycation protein reactions, synergize with sialic acid, and inhibit the formation of glycation protein products by early inhibition, thereby preventing or delaying the non-enzymatic glycation reactions of proteins and the formation of glycation end products.

[0035] To obtain a composition that better inhibits non-enzymatic glycation reactions, the composition also includes carnosine, with the mass ratio of the sialic acid to the carnosine being no greater than 10. Modern pharmacologically active carnosine primarily functions to reduce free radical levels (carnosine is an effective electron donor with metal ion chelating ability, forming complexes with metals and limiting their reactivity); and reacts with protein carbonyl groups (amino groups on carnosine cross-link with amino acid carbonyls or clear modified proteins on macrophages through receptors such as RAGE). Because multiple antioxidant and anti-glycation complex pathways are employed, a better anti-glycation effect can be achieved. The composition may also include excipients such as water, thickeners, and stabilizers that can alter or optimize the composition's morphology.

[0036] To fully utilize the efficacy of the composition containing sialic acid and Poria cocos extract, it can be used to prepare various products that inhibit advanced glycation end products (AGEs), such as anti-glycation products. The products are not limited to pharmaceutical compositions, cosmetics, or special-purpose foods. The dosage forms of the products may include lotions, baths, liniments, tinctures, oils, creams, ointments, plasters, pastes, patches, coatings, films, gels, cataplasms, compresses, sprays, aerosols, implants, emulsions, suppositories, tablets, capsules, pills, powders, and the like.

[0037] In order to ensure the application of the composition containing sialic acid and Poria cocos extract in anti-glycation products, pharmaceutically acceptable excipients and / or carriers, cosmetics or special-purpose foods acceptable excipients and / or carriers may also be included. In this embodiment, acceptable excipients include added surfactants, transdermal absorption enhancers, preservatives, antioxidants, moisturizers, essences, spices and other excipients; acceptable carriers include oily bases or water-soluble bases, oily bases include vaseline, vegetable oils, paraffin, silicone oil, hydrogenated vegetable oils, triglycerides, stearic acid, hexadecanol, acetone, petroleum ether, ethyl oleate, etc.; water-soluble bases include water, distilled water, glycerin, gelatin, xanthan gum, chitosan, methylcellulose, polyethylene glycol, etc.

[0038] Moreover, the effective dosage of the composition containing sialic acid and Poria cocos extract in the preparation of an inhibitor of advanced glycation end products (AGEs) can be 1 wt % to 100 wt %.

[0039] The prepared composition can be used directly as an inhibitor of advanced glycation end products, or it can be compounded with various excipients. The total dose of the advanced glycation end product inhibitor is determined by factors such as cost, regulations, dosage form, and application field, and is not limited here. When the composition containing sialic acid and Poria extract of the present invention is used as an oral anti-glycation product, its form includes, for example, tablets, capsules, powders, granules, liquids, granules, sticks, plates, blocks. Substances, solids, rounds, pastes, ointments, capsules, gelling agents, chewing agents, and adhesives. Among them, tablets, capsules, powders, granules and liquid dosage forms are particularly preferred; among them, liquids can also be PET bottles, cans, bottled beverages; water (hot water), milk, fruit juice, green juice, etc. When the composition containing sialic acid and Poria extract of the present invention is used as an external anti-glycation product, it can be used by applying it to the skin, scalp, etc. Any form can be used, including ointments, creams, gels, lotions, emulsions, soaps, masks, pastes, etc.

[0040] Based on the research results of the anti-glycation effect of single sialic acid and the anti-glycation effect of sialic acid combined with Poria cocos extract, the applicant speculates that, first, sialic acid has free amino groups that can compete with carbohydrates for proteins, acetylate the amino end, and reduce the amount of AGEs; second, the free amino groups in sialic acid can react with the reducing carbonyl groups in Amadori products to form inactive substances, eliminate carbonyl groups, and thus prevent further rearrangement of Amadori products. Therefore, sialic acid inhibits the formation and accumulation of advanced glycation end products (AGEs), thereby inhibiting non-enzymatic glycation reactions. When sialic acid is combined with Poria cocos extract, the antioxidant effect of Poria cocos extract inhibits the formation of early glycated protein products, and synergistically with sialic acid to prevent or delay non-enzymatic glycation reactions and the formation of glycation end products of proteins. Prevents skin dullness and darkening and maintains skin transparency.

[0041] Furthermore, the composition containing sialic acid and pachymaran can improve product stability by inhibiting the glycation reaction of amino acids and proteins contained in the product. By inhibiting glycation reactions, the product's color tone can be stabilized. It can be used as an additive in products such as medicines, cosmetics, and health foods.

[0042] The application of a composition containing sialic acid and Poria cocos extract provided by the present invention in inhibiting non-enzymatic glycosylation reaction is described in detail below with reference to the examples.

[0043] Example 1:

[0044] At room temperature, weigh an appropriate amount of N-acetylneuraminic acid, dissolve it in water, and stir to dissolve evenly to prepare a 0.1% N-acetylneuraminic acid solution.

[0045] Example 2:

[0046] At room temperature, weigh an appropriate amount of N-acetylneuraminic acid, dissolve it in water, and stir to dissolve evenly to prepare a 0.5% N-acetylneuraminic acid solution.

[0047] Example 3:

[0048] At room temperature, weigh an appropriate amount of N-acetylneuraminic acid, dissolve it in water, and stir to dissolve evenly to prepare a 1.0% N-acetylneuraminic acid solution.

[0049] Example 4:

[0050] At room temperature, weigh an appropriate amount of N-acetylneuraminic acid, dissolve it in water, and stir to dissolve evenly to prepare a 5.0% N-acetylneuraminic acid solution.

[0051] Example 5:

[0052] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution 0.7) were weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 0.5% carboxymethyl pachymaran, thereby obtaining composition 5 for inhibiting non-enzymatic glycosylation reaction.

[0053] Example 6:

[0054] At room temperature, weigh appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.7), dissolve them in water, and stir to dissolve them evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 1% carboxymethyl pachymaran, thereby obtaining composition 6 for inhibiting non-enzymatic glycosylation reaction.

[0055] Example 7:

[0056] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.7) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 2% carboxymethyl pachymaran, thereby obtaining composition 7 for inhibiting non-enzymatic glycosylation reaction.

[0057] Example 8:

[0058] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.7) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 4% carboxymethyl pachymaran, thereby obtaining composition 8 for inhibiting non-enzymatic glycosylation reaction.

[0059] Example 9:

[0060] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution 0.7) were weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 8% carboxymethyl pachymaran, thereby obtaining composition 9 for inhibiting non-enzymatic glycosylation reaction.

[0061] Example 10

[0062] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.2) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 4% carboxymethyl pachymaran (degree of substitution is 0.2), thereby obtaining a composition 10 for inhibiting non-enzymatic glycosylation reactions.

[0063] Example 11:

[0064] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.3) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 4% carboxymethyl pachymaran (degree of substitution is 0.3), thereby obtaining a composition 11 for inhibiting non-enzymatic glycosylation reactions.

[0065] Example 12:

[0066] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.5) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 4% carboxymethyl pachymaran (degree of substitution is 0.5), thereby obtaining a composition 12 for inhibiting non-enzymatic glycosylation reactions.

[0067] Example 13:

[0068] At room temperature, appropriate amounts of N-acetylneuraminic acid and carboxymethyl pachymaran (degree of substitution is 0.7) are weighed, dissolved in water, and stirred to dissolve evenly to obtain a mixed solution with a concentration of 1% N-acetylneuraminic acid and 4% carboxymethyl pachymaran (degree of substitution is 0.7), thereby obtaining a composition 13 for inhibiting non-enzymatic glycosylation reactions.

[0069] Example 14:

[0070] At room temperature, appropriate amounts of N-acetylneuraminic acid, carboxymethyl tuckahoe polysaccharide (degree of substitution is 0.7) and carnosine are weighed, dissolved in water, and stirred to dissolve evenly to prepare a mixed solution with a concentration of 1% N-acetylneuraminic acid, 2% carboxymethyl tuckahoe polysaccharide (degree of substitution is 0.7) and 0.5% carnosine, thereby obtaining a composition 14 for inhibiting non-enzymatic glycosylation reactions.

[0071] Examples 1 to 14 were subjected to non-enzymatic glycosylation inhibition tests:

[0072] The specific contents of the test are:

[0073] 1.1 Test samples: N-acetylneuraminic acid crystals, white powder, batch number 2011231, provided by Wuhan Zhongke Optical Valley Green Biotechnology Co., Ltd.; carboxymethyl pachymaran, white powder, purchased from outside.

[0074] 1.2 Test materials: bovine serum albumin (BSA), fructose, disodium hydrogen phosphate dodecahydrate (Na2HPO4·12H2O), sodium dihydrogen phosphate dihydrate (NaH2PO4·2H2O), and vitamin C ethyl ether (Vc ethyl ether).

[0075] 1.3 Experimental equipment: microplate reader, analytical balance, pipette, vortex mixer.

[0076] 1.4 Test method:

[0077] 1.4.1 0.2 mol / L phosphate buffer saline (PBS, pH = 7.3-7.4): Dissolve 7.16 g of Na2HPO4·12H2O in deionized water. Add an appropriate amount of penicillin-streptomycin (for antibacterial purposes) and dilute to 100 mL with deionized water to obtain a 0.2 mol / L Na2HPO4 solution. Dissolve 3.12 g of NaH2PO4·2H2O in deionized water and dilute to 100 mL to obtain a 0.2 mol / L NaH2PO4 solution. Mix 81 mL of the 0.2 mol / L Na2HPO4 solution with 19 mL of the 0.2 mol / L Na2HPO4 solution to obtain a 0.2 mol / L Na2HPO4 solution.

[0078] 1.4.2 Bovine serum albumin solution (BSA solution): Weigh a certain amount of BSA, add PBS to make up to 25 mL, and filter through a 0.45 μm water filter membrane;

[0079] 1.4.3 Fructose solution: Weigh a certain amount of fructose and dilute to 25 mL with PBS;

[0080] 1.4.4 BSA-fructose reaction solution: Mix 20 mL of BSA solution with 20 mL of fructose solution;

[0081] 1.4.5 Test solution: Dilute the sample to 10% with deionized water, and then dilute to the corresponding concentration for testing.

[0082] 1.5 Testing

[0083] The test solutions (test, blank) and controls (test, blank) were incubated at 37°C for 5 days. The fluorescence intensity at 440 nm when excited at 370 nm was measured using a fluorescence spectrophotometer. The AGE formation inhibition rate (%) was calculated according to the following formula.

[0084] The calculation formula of inhibition rate (IR) is: IR (%) = [1-(RFU (sample)-RFU (blank group)) / RFU (negative group)] × 100

[0085] RFU stands for Relative Fluorescence Unit

[0086] The results of the non-enzymatic glycosylation inhibition test were:

[0087] Examples 1-4 are non-enzymatic glycosylation inhibition tests of a single component, N-acetylneuraminic acid. The inhibition rate of 0.1% N-acetylneuraminic acid is 1%; the inhibition rate of 0.5% N-acetylneuraminic acid is 10%; the inhibition rate of 1% N-acetylneuraminic acid is 20%; and the inhibition rate of 5% N-acetylneuraminic acid is 74%.

[0088] Examples 5-9 are combinations of carboxymethyl tuckahoe polysaccharide with a degree of substitution of 0.7 and N-acetylneuraminic acid, with the addition of different mass fractions. The inhibition rate of composition 5 is 23%; the inhibition rate of composition 6 is 30%; the inhibition rate of composition 7 is 38%; the inhibition rate of composition 8 is 44% and the inhibition rate of composition 9 is 48%.

[0089] Examples 10-13 are combinations of carboxymethyl pachymaran and N-acetylneuraminic acid at the same mass fraction with different degrees of substitution. The inhibition rate of composition 10 is 28%; the inhibition rate of composition 11 is 38%; the inhibition rate of composition 12 is 40%; and the inhibition rate of composition 13 is 44%.

[0090] Example 14 is a combination of carnosine, carboxymethyl pachymaran and N-acetylneuraminic acid. The inhibition rate of composition 14 is 52%.

[0091] like Figure 1 As shown, this is a non-enzymatic glycation inhibition test of a single ingredient, N-acetylneuraminic acid. The production of AGEs is significantly inhibited. The results show that N-acetylneuraminic acid has a significant non-enzymatic glycation inhibition effect. As its concentration increases, the effect of inhibiting the production of AGEs becomes more obvious. When the concentration is 5%, the inhibition rate is as high as 56%.

[0092] like Figure 2 As shown, the non-enzymatic glycation inhibition test of the combination of N-acetylneuraminic acid and different concentrations of carboxymethyl tuckahoe polysaccharide, compared with the non-enzymatic glycation inhibition effect of a single component N-acetylneuraminic acid, as the concentration of carboxymethyl tuckahoe polysaccharide increases, the effect of inhibiting the formation of AGEs is significantly improved.

[0093] like Figure 3 The figure shows the non-enzymatic glycosylation inhibition test of the composition of N-acetylneuraminic acid and carboxymethyl pachymaran with different substitution degrees. The results show that as the substitution degree increases, the effect of inhibiting the formation of AGEs becomes better.

[0094] like Figure 4 As shown in the figure, the compound N-acetylneuraminic acid, carboxymethyl tuckahoe polysaccharide and carnosine can also effectively inhibit the formation of AGEs, and the multiple antioxidant and anti-glycation complex pathways can achieve better anti-glycation effects.

[0095] In summary, because non-enzymatic saccharification reaction refers to the process in which the aldehyde or ketone groups of reducing sugars and substances containing free amino groups such as proteins and lipids first form a reversible Schiff base under the conditions of no enzyme catalysis, and then through reactions such as rearrangement, oxidation and dehydration, stable characteristic fluorescent advanced glycation end products (AGEs) are finally formed. Proteins undergo cross-linking, browning and fluorescence generation through repeated glycosylation, which can promote protein function reduction and aging. N-acetylneuraminic acid provided by the present invention is a significant AGEs inhibitor. After compounding carboxymethyl tuckahoe polysaccharide, it is more effective in improving the inhibition of AGEs generation. This is because the antioxidant activity of carboxymethyl tuckahoe polysaccharide itself has a synergistic effect on N-acetylneuraminic acid, and the antioxidant activity of carboxymethyl tuckahoe polysaccharide can inhibit the formation of early fructosamine and dicarbonyl compounds, thereby collaborating with N-acetylneuraminic acid to further inhibit the formation of glycation end products.

[0096] 2. Using the composition containing sialic acid and Poria cocos extract to prepare anti-glycation products

[0097] 2.1 Tablets

[0098] Conventional excipients are added to the composition containing sialic acid and Poria cocos extract of the present invention, and tablets are prepared using conventional methods.

[0099] 2.2 Granules

[0100] Conventional auxiliary materials are added to the composition containing sialic acid and Poria cocos extract of the present invention, and granules are prepared using conventional methods.

[0101] 2.3 Capsules

[0102] Conventional excipients are added to the composition containing sialic acid and Poria cocos extract of the present invention, and capsules are prepared using conventional methods.

[0103] 2.4 Liquid

[0104] Conventional excipients are added to the composition containing sialic acid and Poria cocos extract of the present invention, and a liquid preparation is prepared by a conventional method.

[0105] 2.5 Lotion

[0106] Conventional auxiliary materials are added to the composition containing sialic acid and Poria cocos extract of the present invention, and an emulsion is prepared by a conventional method.

[0107] 2.6 Beauty Cream

[0108] The composition containing sialic acid and Poria cocos extract of the present invention is added with conventional auxiliary materials and a conventional method is used to prepare a beauty cream.

[0109] In the absence of conflict, the above embodiments and features in the embodiments may be combined with each other.

[0110] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition for inhibiting non-enzymatic glycosylation reaction, characterized in that: The composition comprises N-acetylneuraminic acid and carboxymethyl tuckahoe polysaccharide, wherein the mass concentration ratio of N-acetylneuraminic acid to carboxymethyl tuckahoe polysaccharide in the composition is 1:(4-8), and the degree of substitution of the carboxymethyl tuckahoe polysaccharide is 0.5-0.

7.

2. The composition according to claim 1, characterized in that Carnosine is also included in the composition.

3. The composition according to claim 2, characterized in that The mass concentration ratio of N-acetylneuraminic acid, carboxymethyl pachymaran and carnosine in the composition is 1:2:0.

5.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of an advanced glycosylation end product (AGE) inhibitor.

5. The use according to claim 4, characterized in that: The addition amount of the composition in the preparation of advanced glycosylation end products AGEs inhibitor is 1-100wt%.

6. A cosmetic, characterized in that: The cosmetic contains the composition according to any one of claims 1 to 3.

7. An anti-glycation product, characterized in that: The anti-glycation product contains the composition according to any one of claims 1 to 3.

8. The anti-glycation product according to claim 7, characterized in that Pharmaceutically acceptable excipients are also included.

9. The anti-glycation product according to claim 8, characterized in that The dosage form of the anti-glycation product includes any one of tablets, granules, capsules or liquids.

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