Compound amino acid composition for enhancing telomerase activity and preparation method thereof

Abstract

The invention relates to the technical field of amino acid compositions, and discloses a compound amino acid composition for enhancing telomerase activity, which comprises the following components (by weight): 20-25 parts of L-arginine; 15 to 20 parts of L-lysine; 25 to 30 parts of L-glutamine; 10 to 15 parts of L-serine; 5 to 8 parts of L-glycine; 3 to 5 parts of L-proline; 2 to 4 parts of N-acetyl-L-cysteine; 1 to 2 parts of L-tryptophan; 1 to 2 parts of beta-alanine; 2 to 3 parts of L-carnitine; 1-2 parts of taurine; through the synergistic effect of multiple mechanisms, the compound amino acid composition not only significantly enhances the telomerase activity (up to 195%), but also can maintain the telomere length, inhibit cell aging and provide antioxidant protection. The comprehensive regulation effect is derived from a unique metabolic network formed among the components, and mutual promotion and regulation are realized. Experimental results show that the long telomere length (8.3 kb vs control group 7.1 kb) can be maintained in 30-day long-term culture.

Description

Technical Field

[0001] The invention relates to the technical field of composite amino acid compositions, in particular to a composite amino acid composition for enhancing telomerase activity and a preparation method thereof. Background Art

[0002] As a special reverse transcriptase, telomerase plays a key role in maintaining cell telomere length and delaying cell aging. In recent years, with the intensification of the problem of population aging, the regulation of telomerase activity has become a hot area in anti-aging research. At present, the research on telomerase activity enhancers is mainly focused on the following directions:

[0003] 1. Small molecule activators: such as TA-65 and cycloastragenol, these compounds can enhance telomerase activity by activating the expression of the telomerase gene hTERT. However, these small molecule compounds often have problems such as low bioavailability and unknown safety for long-term use.

[0004] 2. Gene therapy: The activity of telomerase is enhanced by introducing the hTERT gene. However, this method has the problems of complicated operation and potential carcinogenicity.

[0005] 3. Proteins and peptides: such as LPTS protein and its derivatives, which can directly bind to telomerase and regulate its activity. However, large molecular proteins are difficult to enter cells and are easily degraded by the body, which limits their application.

[0006] 4. Single amino acid supplementation: such as arginine, lysine, etc. Although some amino acids can affect telomerase activity to a certain extent, the effect is limited and unstable.

[0007] The main problems of the existing technology:

[0008] 1. Single effect: Most existing technologies only enhance telomerase activity from a single perspective, ignoring the complexity and multifactorial nature of cell aging.

[0009] 2. Safety risks: Some telomerase activators may cause excessive extension of telomeres and increase the risk of cancer.

[0010] 3. Low bioavailability: Many active substances are difficult to be absorbed and utilized by the human body, which limits their actual effects.

[0011] 4. Insufficient long-term effectiveness: Some telomerase activators can only increase telomerase activity in the short term and it is difficult to maintain long-term effects.

[0012] 5. Lack of synergistic effect: Single-ingredient telomerase activators are difficult to fully regulate cell metabolism and anti-aging processes.

[0013] 6. Improper preparation process: Certain preparation methods may cause the inactivation of active ingredients or produce harmful by-products.

[0014] In view of the above problems, there is an urgent need to develop a safe, efficient and long-term effective telomerase activity enhancer to meet the growing anti-aging needs. Summary of the invention

[0015] 1. Technical issues to be resolved

[0016] In view of the deficiencies of the prior art, the present invention provides a composite amino acid composition for enhancing telomerase activity and a preparation method thereof.

[0017] (II) Technical solution

[0018] To achieve the above object, the present invention provides the following technical solution: a composite amino acid composition for enhancing telomerase activity and a preparation method thereof.

[0019] A composite amino acid composition for enhancing telomerase activity, the composition comprising the following components (in parts by weight):

[0020] L-Arginine 20-25 parts

[0021] 15-20 parts of L-lysine

[0022] L-Glutamine 25-30 parts

[0023] L-serine 10-15 parts

[0024] 5-8 parts of L-glycine

[0025] 3-5 parts of L-proline

[0026] 2-4 parts of N-acetyl-L-cysteine

[0027] 1-2 servings of L-Tryptophan

[0028] 1-2 parts of β-alanine

[0029] 2-3 servings of L-Carnitine

[0030] 1-2 servings of Taurine

[0031] Preferably, the amount of L-arginine is 22-24 parts.

[0032] Preferably, the amount of L-glycine used is 6-7 parts.

[0033] Preferably, the amount of N-acetyl-L-cysteine ​​is 2.5-3.5 parts.

[0034] Preferably, the composition further comprises one or more vitamins.

[0035] A method for preparing the composition comprises the following steps:

[0036] (1) Mix all amino acids in proportion and add 10 times the weight of purified water;

[0037] (2) adjusting the pH to 7.2-7.4 and adding compound protease;

[0038] (3) mild enzymatic hydrolysis at 37°C for 6-8 hours, keeping the pH constant;

[0039] (4) heating the enzymatic solution to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature;

[0040] (5) ultrafiltration of the obtained solution to remove macromolecular proteins and enzymes;

[0041] (6) The ultrafiltered solution is freeze-dried to obtain the final product.

[0042] Preferably, the composite protease comprises trypsin, papain and proteinase K, wherein the weight ratio of trypsin:papain:proteinase K is 2:1:1.

[0043] Preferably, the enzymatic hydrolysis time in step (3) is 7 hours.

[0044] Preferably, the ultrafiltration in step (5) uses an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons.

[0045] Preferably, the conditions for low-temperature freeze drying in step (6) are: pre-freezing temperature -40°C, vacuum degree less than 10 Pa, and drying temperature not exceeding 25°C.

[0046] (III) Beneficial technical effects

[0047] Compared with the prior art, the present invention has the following beneficial technical effects:

[0048] 1. Multiple synergistic effects: The composite amino acid composition of the present invention works synergistically through multiple mechanisms, not only significantly enhancing telomerase activity (up to 195%), but also maintaining telomere length, inhibiting cell aging and providing antioxidant protection. This comprehensive regulatory effect stems from the unique metabolic network formed between the components, which promote and regulate each other.

[0049] 2. Long-term telomere protection: Experimental results show that the present invention can maintain a longer telomere length (8.3kb vs. 7.1kb in the control group) in a 30-day long-term culture. This sustained protective effect may be the result of the combined effects of L-proline regulating telomerase activity through the p53 signaling pathway and N-acetyl-L-cysteine ​​providing antioxidant protection.

[0050] 3. Significant anti-aging effect: The present invention significantly reduced the proportion of SA-β-gal positive cells (12.8% vs. 28.6% in the control group), indicating that it has strong anti-aging ability. This effect may be due to the fact that the present invention not only enhances telomerase activity, but also comprehensively improves the cell state through multiple pathways such as L-glycine providing methyl donors and L-tryptophan regulating NAD+ metabolism.

[0051] 4. Excellent antioxidant capacity: The present invention exhibits a DPPH free radical scavenging rate of 52.3%, which is much higher than that of single amino acids or traditional antioxidants. This strong antioxidant capacity may be produced by the synergistic effect of L-glycine, N-acetyl-L-cysteine ​​and L-tryptophan, which not only directly scavenges free radicals, but also may play a role by regulating the intracellular antioxidant enzyme system.

[0052] 5. High bioavailability: The present invention adopts a mild enzymatic hydrolysis process, which not only retains the biological activity of amino acids, but also may produce small molecule peptides with unique biological functions. This process innovation significantly improves the bioavailability of the product, making it easier for the human body to absorb and utilize it.

[0053] 6. Safety advantage: The present invention uses a combination of natural amino acids, avoiding the potential safety hazards of a single powerful activator. It gently regulates telomerase activity through multiple mechanisms, reducing the risk of over-activation.

[0054] 7. Personalized adjustment potential: The multi-component formula of the present invention provides a basis for the future development of personalized anti-aging solutions, and the amino acid ratio can be adjusted according to different individual needs.

[0055] In summary, the present invention achieves significant enhancement of telomerase activity and provides comprehensive cell protection through innovative composite amino acid combination and optimized preparation process. This multiple synergistic effect not only solves many problems existing in the prior art, but also shows excellent anti-aging and antioxidant effects, providing innovative ideas and scientific basis for the development of a new generation of anti-aging products. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 is a bar graph of telomerase activity assay results;

[0057] Figure 2 is a bar graph of telomerase activity assay results;

[0058] Figure 3 is a bar graph of cell senescence measurement results;

[0059] Figure 4 It is a bar graph of the antioxidant capacity test results. DETAILED DESCRIPTION

[0060] Please refer to Figure 1-4 To achieve the above purpose, the present invention provides the following implementation modes and embodiments:

[0061] Example 1

[0062] A composite amino acid composition for enhancing telomerase activity, the components of which are as follows (by weight):

[0063] L-Arginine 20 servings

[0064] 15 parts of L-Lysine

[0065] L-Glutamine 25 parts

[0066] 10 parts of L-serine

[0067] 5 parts of L-Glycine

[0068] 3 parts of L-proline

[0069] 2 parts N-acetyl-L-cysteine

[0070] 1 portion of L-Tryptophan

[0071] 1 part of β-alanine

[0072] 2 parts L-Carnitine

[0073] 1 part Taurine

[0074] The preparation method of the composite amino acid composition comprises the following steps:

[0075] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1000 parts) was added;

[0076] (2) adjusting the pH to 7.2 with 1 mol / L sodium hydroxide solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:100;

[0077] (3) Perform mild enzymatic hydrolysis at 37°C for 6 h, check the pH every 30 min, and adjust the pH to 7.2 with 1 mol / L sodium hydroxide solution if necessary;

[0078] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 20°C) in an ice water bath;

[0079] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0080] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 5 Pa, the drying temperature is 20°C, and the drying time is 48 hours, thereby obtaining the final product.

[0081] Example 2

[0082] A composite amino acid composition for enhancing telomerase activity, the components of which are as follows (by weight):

[0083] 25 servings of L-arginine

[0084] 20 servings of L-Lysine

[0085] L-Glutamine 30 servings

[0086] 15 parts of L-serine

[0087] 8 parts of L-Glycine

[0088] 5 parts of L-proline

[0089] 4 parts of N-acetyl-L-cysteine

[0090] 2 parts of L-Tryptophan

[0091] 2 parts of β-alanine

[0092] 3 parts of L-Carnitine

[0093] 2 parts Taurine

[0094] The preparation method of the composite amino acid composition comprises the following steps:

[0095] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1160 parts) was added;

[0096] (2) adjusting the pH to 7.4 with 1 mol / L hydrochloric acid solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:50;

[0097] (3) Perform mild enzymatic hydrolysis at 37°C for 8 h, check the pH every 60 min, and adjust the pH to 7.4 with 1 mol / L hydrochloric acid solution if necessary;

[0098] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 22°C) in an ice water bath;

[0099] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0100] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 10 Pa, the drying temperature is 25°C, and the drying time is 72 hours, thereby obtaining the final product.

[0101] Example 3

[0102] A composite amino acid composition for enhancing telomerase activity, the components of which are as follows (by weight):

[0103] 22 servings of L-arginine

[0104] 18 parts of L-Lysine

[0105] L-Glutamine 28 parts

[0106] 12 servings of L-serine

[0107] 6 parts of L-Glycine

[0108] 4 parts of L-proline

[0109] 3 parts of N-acetyl-L-cysteine

[0110] 1.5 parts of L-Tryptophan

[0111] 1.5 parts of β-alanine

[0112] 2.5 parts of L-Carnitine

[0113] 1.5 parts of Taurine

[0114] The preparation method of the composite amino acid composition comprises the following steps:

[0115] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1000 parts) was added;

[0116] (2) adjusting the pH to 7.3 with 0.5 mol / L sodium hydroxide solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:75;

[0117] (3) Perform mild enzymatic hydrolysis at 37°C for 7 h, check the pH every 45 min, and adjust the pH to 7.3 with 0.5 mol / L sodium hydroxide solution if necessary;

[0118] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 21°C) in an ice water bath;

[0119] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0120] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 7 Pa, the drying temperature is 22°C, and the drying time is 60 hours, thereby obtaining the final product.

[0121] Example 4

[0122] A composite amino acid composition for enhancing telomerase activity, the components of which are as follows (by weight):

[0123] 24 servings of L-arginine

[0124] 17 parts of L-Lysine

[0125] L-Glutamine 27 parts

[0126] 14 parts of L-serine

[0127] 7 parts of L-Glycine

[0128] 3.5 parts of L-proline

[0129] 3.5 parts of N-acetyl-L-cysteine

[0130] 1.8 parts of L-Tryptophan

[0131] 1.2 parts of β-alanine

[0132] 2.8 parts of L-Carnitine

[0133] Taurine 1.2 parts

[0134] The preparation method of the composite amino acid composition comprises the following steps:

[0135] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1030 parts) was added;

[0136] (2) adjusting the pH to 7.3 with 0.1 mol / L phosphate buffer, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:60;

[0137] (3) Perform mild enzymatic hydrolysis at 37°C for 7.5 hours, check the pH every 40 minutes, and adjust the pH to 7.3 with 0.1 mol / L phosphate buffer if necessary;

[0138] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 23°C) in an ice water bath;

[0139] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0140] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 8 Pa, the drying temperature is 23°C, and the drying time is 54 hours, thereby obtaining the final product.

[0141] Comparative Example 1 (corresponding to Example 1, verifying the importance of L-glycine):

[0142] Components (by weight):

[0143] L-Arginine 20 servings

[0144] 15 parts of L-Lysine

[0145] L-Glutamine 25 parts

[0146] 10 parts of L-serine

[0147] 3 parts of L-proline

[0148] 2 parts N-acetyl-L-cysteine

[0149] 1 portion of L-Tryptophan

[0150] 1 part of β-alanine

[0151] 2 parts L-Carnitine

[0152] 1 part Taurine

[0153] Preparation method:

[0154] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 800 parts) was added;

[0155] (2) adjusting the pH to 7.2 with 1 mol / L sodium hydroxide solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:100;

[0156] (3) Perform mild enzymatic hydrolysis at 37°C for 6 h, check the pH every 30 min, and adjust the pH to 7.2 with 1 mol / L sodium hydroxide solution if necessary;

[0157] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 20°C) in an ice water bath;

[0158] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0159] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 5 Pa, the drying temperature is 20°C, and the drying time is 48 hours, thereby obtaining the final product.

[0160] Comparative Example 2 (corresponding to Example 2, verifying the importance of N-acetyl-L-cysteine):

[0161] Components (by weight):

[0162] 25 servings of L-arginine

[0163] 20 servings of L-Lysine

[0164] L-Glutamine 30 servings

[0165] 15 parts of L-serine

[0166] 8 parts of L-Glycine

[0167] 5 parts of L-proline

[0168] 2 parts of L-Tryptophan

[0169] 2 parts of β-alanine

[0170] 3 parts of L-Carnitine

[0171] 2 parts Taurine

[0172] Preparation method:

[0173] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1120 parts) was added;

[0174] (2) adjusting the pH to 7.4 with 1 mol / L hydrochloric acid solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:50;

[0175] (3) Perform mild enzymatic hydrolysis at 37°C for 8 h, check the pH every 60 min, and adjust the pH to 7.4 with 1 mol / L hydrochloric acid solution if necessary;

[0176] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 22°C) in an ice water bath;

[0177] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0178] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 10 Pa, the drying temperature is 25°C, and the drying time is 72 hours, thereby obtaining the final product.

[0179] Comparative Example 3 (corresponding to Example 3, verifying the importance of L-arginine content):

[0180] Components (by weight):

[0181] L-Arginine 10 parts (reduced content)

[0182] 18 parts of L-Lysine

[0183] L-Glutamine 28 parts

[0184] 12 servings of L-serine

[0185] 6 parts of L-Glycine

[0186] 4 parts of L-proline

[0187] 3 parts N-acetyl-L-cysteine

[0188] 1.5 parts of L-Tryptophan

[0189] 1.5 parts of β-alanine

[0190] 2.5 parts of L-Carnitine

[0191] 1.5 parts of Taurine

[0192] Preparation method:

[0193] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 880 parts) was added;

[0194] (2) adjusting the pH to 7.3 with 0.5 mol / L sodium hydroxide solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:75;

[0195] (3) Perform mild enzymatic hydrolysis at 37°C for 7 h, check the pH every 45 min, and adjust the pH to 7.3 with 0.5 mol / L sodium hydroxide solution if necessary;

[0196] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 21°C) in an ice water bath;

[0197] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0198] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 7 Pa, the drying temperature is 22°C, and the drying time is 60 hours, thereby obtaining the final product.

[0199] Comparative Example 4 (corresponding to Example 4, verifying the importance of L-glutamine content):

[0200] Components (by weight):

[0201] 24 servings of L-arginine

[0202] 17 parts of L-Lysine

[0203] L-Glutamine 40 parts (to increase content)

[0204] 14 parts of L-serine

[0205] 7 parts of L-Glycine

[0206] 3.5 parts of L-proline

[0207] 3.5 parts of N-acetyl-L-cysteine

[0208] 1.8 parts of L-Tryptophan

[0209] 1.2 parts of β-alanine

[0210] 2.8 parts of L-Carnitine

[0211] Taurine 1.2 parts

[0212] Preparation method:

[0213] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1160 parts) was added;

[0214] (2) adjusting the pH to 7.3 with 0.1 mol / L phosphate buffer, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:60;

[0215] (3) Perform mild enzymatic hydrolysis at 37°C for 7.5 hours, check the pH every 40 minutes, and adjust the pH to 7.3 with 0.1 mol / L phosphate buffer if necessary;

[0216] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 23°C) in an ice water bath;

[0217] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0218] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 8 Pa, the drying temperature is 23°C, and the drying time is 54 hours, thereby obtaining the final product.

[0219] Comparative Example 5 (corresponding to Example 1, verifying the importance of L-proline):

[0220] Components (by weight):

[0221] L-Arginine 20 servings

[0222] 15 parts of L-Lysine

[0223] L-Glutamine 25 parts

[0224] 10 parts of L-serine

[0225] 5 parts of L-Glycine

[0226] 3 servings of L-Leucine (substitute for L-Proline)

[0227] 2 parts N-acetyl-L-cysteine

[0228] 1 portion of L-Tryptophan

[0229] 1 part of β-alanine

[0230] 2 parts L-Carnitine

[0231] 1 part Taurine

[0232] Preparation method:

[0233] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 850 parts) was added;

[0234] (2) adjusting the pH to 7.2 with 1 mol / L sodium hydroxide solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:100;

[0235] (3) Perform mild enzymatic hydrolysis at 37°C for 6 h, check the pH every 30 min, and adjust the pH to 7.2 with 1 mol / L sodium hydroxide solution if necessary;

[0236] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 20°C) in an ice water bath;

[0237] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0238] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 5 Pa, the drying temperature is 20°C, and the drying time is 48 hours, thereby obtaining the final product.

[0239] Comparative Example 6 (corresponding to Example 2, verifying the importance of enzymolysis time):

[0240] Components (by weight):

[0241] 25 servings of L-arginine

[0242] 20 servings of L-Lysine

[0243] L-Glutamine 30 servings

[0244] 15 parts of L-serine

[0245] 8 parts of L-Glycine

[0246] 5 parts of L-proline

[0247] 4 parts of N-acetyl-L-cysteine

[0248] 2 parts of L-Tryptophan

[0249] 2 parts of β-alanine

[0250] 3 parts of L-Carnitine

[0251] 2 parts Taurine

[0252] Preparation method:

[0253] (1) All amino acids were mixed in proportion and 10 times the weight of purified water (i.e. 1160 parts) was added;

[0254] (2) adjusting the pH to 7.4 with 1 mol / L hydrochloric acid solution, adding a composite protease (trypsin: papain: proteinase K = 2:1:1), wherein the ratio of enzyme to substrate is 1:50;

[0255] (3) Perform mild enzymatic hydrolysis at 37°C for 4 h (shorten the enzymatic hydrolysis time), check the pH every 30 min, and adjust the pH to 7.4 with 1 mol / L hydrochloric acid solution if necessary;

[0256] (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature (about 22°C) in an ice water bath;

[0257] (5) ultrafiltration of the obtained solution using an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons to remove macromolecular proteins and enzymes;

[0258] (6) The ultrafiltered solution is subjected to low-temperature freeze drying, wherein the pre-freezing temperature is -40°C, the vacuum degree is 10 Pa, the drying temperature is 25°C, and the drying time is 72 hours, thereby obtaining the final product.

[0259] Based on the above embodiments and comparative examples, the applicant designed multiple pharmacological models to evaluate the effectiveness of the scheme and conducted a detailed analysis of the experimental results.

[0260] 1. Telomerase activity assay model (TRAP method)

[0261] Experimental conditions: Telomerase activity was determined using the TRAP (Telomeric Repeat Amplification Protocol) method.

[0262] Experimental steps:

[0263] (1) Telomerase was extracted from HEK293 cells.

[0264] (2) Each component and comparative example sample is reacted with telomerase.

[0265] (3) Amplify the reaction product by PCR.

[0266] (4) Separate the products by polyacrylamide gel electrophoresis.

[0267] (5) Telomerase activity was quantitatively analyzed using a phosphor imaging system.

[0268] 2. Telomere length determination model (TRF method)

[0269] Experimental conditions: Telomere length was determined using telomere restriction fragment (TRF) analysis.

[0270] Experimental steps:

[0271] (1) Human fibroblasts were cultured and subcultured every 3 days for a total of 30 days.

[0272] (2) Extract cell DNA and digest it with restriction endonucleases.

[0273] (3) Telomere length was detected by Southern blot method.

[0274] (4) Telomere length was analyzed using ImageJ software.

[0275] 3. Cell senescence model (SA-β-gal staining method)

[0276] Experimental conditions: SA-β-galactosidase staining was used to detect the degree of cell senescence. Experimental steps:

[0277] (1) Human fibroblasts were cultured and passaged every 3 days for 30 days. (2) The cells were fixed and stained with SA-β-gal.

[0278] (3) Count the proportion of positive cells under a microscope.

[0279] 4. Antioxidant capacity determination model (DPPH free radical scavenging method)

[0280] Experimental conditions: The antioxidant capacity was determined using the DPPH free radical scavenging method.

[0281] Experimental steps:

[0282] (1) Mix the sample with DPPH solution.

[0283] (2) Incubate in dark place for 30 minutes.

[0284] (3) Measure the absorbance at 517 nm.

[0285] (4) Calculate the DPPH free radical scavenging rate.

[0286] The experimental results are as follows:

[0287] Table 1. Telomerase activity assay results (relative activity, with the control group as 100%)

[0288]

[0289]

[0290] Table 2. Telomere length measurement results (kb)

[0291]

[0292]

[0293] Table 3. Cell senescence assay results (percentage of SA-β-gal positive cells)

[0294]

[0295]

[0296] Table 4. Antioxidant capacity determination results (DPPH free radical scavenging rate)

[0297] sample DPPH free radical scavenging rate (%) Control group 12.5±1.2 Example 1 45.6±3.5 Example 2 52.3±4.0 Example 3 48.7±3.8 Example 4 50.9±3.9 Comparative Example 1 32.1±2.8 Comparative Example 2 28.4±2.5 Comparative Example 3 35.6±3.0 Comparative Example 4 38.2±3.2 Comparative Example 5 30.8±2.7 Comparative Example 6 34.5±3.0

[0298] Best embodiment: According to the comprehensive test results, Example 2 shows the best telomerase activity enhancement effect, telomere length maintenance ability, cell aging inhibition effect and antioxidant ability.

[0299] Unexpected technical effects:

[0300] 1. Synergistic effect: The composite amino acid composition of the present invention not only significantly enhances telomerase activity (up to 195%), but also shows excellent effects in maintaining telomere length and inhibiting cell aging. This synergistic effect may be due to the formation of a unique metabolic network between the components, which promote and regulate each other.

[0301] 2. Long-term telomere protection effect: Example 2 can still maintain a longer telomere length (8.3 kb vs. 7.1 kb in the control group) during 30 days of long-term culture, which shows that the present invention can not only activate telomerase in the short term, but also protect telomeres in the long term, which may be related to the synergistic effect of L-proline and N-acetyl-L-cysteine.

[0302] 3. Significant anti-aging effect: The proportion of SA-β-gal positive cells in Example 2 after 30 days was only 12.8%, far lower than 28.6% of the control group. This strong anti-aging effect may be due to the fact that the present invention not only enhances telomerase activity, but also provides a comprehensive cell protection mechanism.

[0303] 4. Excellent antioxidant capacity: Example 2 exhibited a DPPH free radical scavenging rate of 52.3%, which was much higher than that of single amino acids or traditional antioxidants. This strong antioxidant capacity may be produced by the synergistic effect of L-glycine, N-acetyl-L-cysteine ​​and L-tryptophan.

[0304] 5. The key role of the preparation process: The results of Comparative Example 6 show that even if the components are the same, shortening the enzymatic hydrolysis time will significantly reduce the effect of the product. This highlights the importance of the mild enzymatic hydrolysis process in the present invention, which may be because this process can better retain the biological activity of amino acids and produce beneficial small molecule peptides.

[0305] These unexpected technical effects are mainly due to the following reasons:

[0306] 1. Multiple mechanisms of action: The composite amino acid composition of the present invention acts synergistically through multiple pathways, including directly activating telomerase, providing the raw materials required for telomere DNA synthesis, regulating telomerase gene expression, enhancing antioxidant defense, etc.

[0307] 2. Synergistic effect of key components: L-glycine provides a methyl donor, L-proline regulates the p53 signaling pathway, N-acetyl-L-cysteine ​​enhances antioxidant capacity, and L-tryptophan affects NAD+ metabolism. These components work together to produce effects beyond expectations.

[0308] 3. Optimized component ratio: By precisely regulating the ratio of each amino acid, a microenvironment most suitable for telomerase to function is created, while balancing the overall metabolic state of the cells.

[0309] 4. Innovative preparation process: The mild enzymatic hydrolysis process not only retains the biological activity of amino acids, but also may produce small molecule peptides with unique biological functions, further enhancing the overall effect of the product.

[0310] In summary, the present invention achieves a significant enhancement of telomerase activity through the synergistic effect of multiple mechanisms, while providing comprehensive cell protection, showing excellent anti-aging and antioxidant effects, and providing innovative ideas for the development of a new generation of anti-aging products.

Claims

1. A composite amino acid composition for enhancing telomerase activity, characterized in that , the composition comprises the following components (in parts by weight): L-Arginine 20-25 parts 15-20 parts of L-lysine L-Glutamine 25-30 parts L-serine 10-15 parts 5-8 parts of L-glycine 3-5 parts of L-proline 2-4 parts of N-acetyl-L-cysteine 1-2 servings of L-Tryptophan 1-2 parts of β-alanine 2-3 servings of L-Carnitine Taurine 1-2 parts.

2. The composition according to claim 1, characterized in that , the amount of L-arginine used is 22-24 parts.

3. The composition according to claim 1, characterized in that , the amount of L-glycine used is 6-7 parts.

4. The composition according to claim 1, characterized in that , the dosage of the N-acetyl-L-cysteine ​​is 2.5-3.5 parts.

5. The composition according to any one of claims 1 to 4, characterized in that , the composition also includes one or more vitamins.

6. A method for preparing the composition according to any one of claims 1 to 5, characterized in that , including the following steps: (1) Mix all amino acids in proportion and add 10 times the weight of purified water; (2) Adjust the pH to 7.2-7.4 and add compound protease; (3) Perform mild enzymatic hydrolysis at 37°C for 6-8 hours, keeping the pH constant; (4) heating the enzymatic hydrolysate to 80°C and maintaining it for 10 minutes to inactivate the enzyme, and then rapidly cooling it to room temperature; (5) ultrafiltration of the obtained solution to remove macromolecular proteins and enzymes; (6) The ultrafiltered solution is freeze-dried to obtain the final product.

7. The method according to claim 6, characterized in that The composite protease comprises trypsin, papain and proteinase K, wherein the weight ratio of trypsin:papain:proteinase K is 2:1:

1.

8. The method according to claim 6, characterized in that , the enzymatic hydrolysis time in step (3) is 7 hours.

9. The method according to claim 6, characterized in that The ultrafiltration in step (5) uses an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons.

10. The method according to claim 6, characterized in that The conditions for low-temperature freeze-drying in step (6) are: pre-freezing temperature -40°C, vacuum degree less than 10Pa, and drying temperature not exceeding 25°C.

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