WO2023216952A1

WO2023216952A1 - Anti-ageing use of uridylic acid, adenylic acid, and yeast peptide composition

Info

Publication number: WO2023216952A1
Application number: PCT/CN2023/091948
Authority: WO
Inventors: 李勇; 徐美虹; 陈玉松; 曾峥
Original assignee: 陈玉松
Priority date: 2022-05-09
Filing date: 2023-05-04
Publication date: 2023-11-16
Also published as: CN114931625A

Abstract

Provided is an anti-ageing use of a uridylic acid, adenylic acid, and yeast peptide composition, relating to the technical field of medicine and healthcare. The composition comprises 5'-uridylic acid, 5'-adenylic acid, and yeast peptide, and has an anti-ageing function. Provided is the anti-ageing use of a composition comprising the three components of 5'-uridylic acid, 5'- adenylic acid, and yeast peptide, wherein the three components can promote the absorption of one another in the intestinal tract, and the 5'-uridylic acid and 5'-adenylic acid and the yeast peptide can play a synergistic role in cells and tissues, effectively removing DPPH and OH-free radicals, repairing damaged cells, and increasing the activity of antioxidant enzymes, thereby achieving the purpose of delaying and preventing ageing.

Description

Anti-aging applications of uridylic acid, adenylic acid and yeast peptide compositions

Technical field

The invention belongs to the technical field of medicine and health care, and specifically relates to the application of uridylic acid, adenylic acid and yeast peptide compositions in anti-aging.

Background technique

my country has entered a society with an aging population since the beginning of the 21st century. More than 20 years have passed. The aging of the population in our country continues to deepen, and it is about to enter a moderately aging society. Data from the seventh national census show that as of 0:00 on November 1, 2020, my country's elderly population aged 60 and above reached 264 million, accounting for approximately 18.70% of the total population, and the population aged 65 and above reached 190 million. Accounting for about 13.50% of the total population. Compared with the fifth national census data in 2000, the proportion of the population aged 60 and above and the proportion of the population aged 65 and above increased by 8.6 percentage points and 6.5 percentage points respectively. And it is expected to be by 2035. Our country is about to enter a severely aging society. With the deepening of population aging, delaying aging has gradually become one of the urgent solutions under the current situation.

Delaying aging can literally be understood as delaying the aging of the body, including the aging of cells, tissues, organs, etc. However, with the development of people's cognition, we have a more comprehensive understanding of delaying aging. Anything that is beneficial to human health and improves the quality of life can be regarded as delaying aging. The main ways to delay aging are non-drug and drug. The non-drug method is to adhere to scientific life rules and maintain good living habits. There are many categories of drug methods, most of which are health-care drugs or foods, and their effects are also the most significant.

Nucleotides are the basic building blocks of ribonucleic acid and deoxyribonucleic acid, and are the precursors of nucleic acid synthesis in the body. Nucleotides are distributed along with nucleic acids in the nuclei and cytoplasm of various organs, tissues, and cells in organisms, and as components of nucleic acids, they participate in basic life activities such as inheritance, development, and growth of organisms.

Contents of the invention

The object of the present invention is to provide an anti-aging application of a composition of uridylic acid, adenylic acid and yeast peptide. After long-term research, this invention has found that different nucleotides have different functions. When combined with yeast peptide, it can effectively remove free radicals in the body, delay cell aging, and restore youthfulness. The present invention provides a new idea for the application of nucleotide nutritional products in improving human health and people's quality of life. The obtained finished product has the effect of delaying the aging of the body and restoring youthfulness.

In order to achieve the above objects, the present invention is achieved through the following technical solutions:

A composition of uridylic acid, adenylic acid and yeast peptide, the composition includes 5'-uridylic acid, 5'-adenylic acid and yeast peptide, and has anti-aging function.

Further, in the above technical solution, the composition includes components in the following weight ratio: 5'-uridylic acid 14-63%, 5'-adenosine 9-50%, yeast peptide 1.5-77%.

Further, in the above technical solution, the composition includes components in the following weight ratio: 5’-uridylic acid 40-60%, 5’-adenylic acid 25-50%, and yeast peptide 10-30%.

Further, in the above technical solution, the composition includes components in the following weight ratio: 5'-uridylic acid 41.5%, 5'-adenylic acid 41.5%, yeast peptide 17%; or 5'-uridine Acid 52%, 5'-adenylate 26%, yeast peptide 22%.

Further, in the above technical solution, 5'-uridylic acid and 5'-adenylic acid exist in the form of 5'-monophosphate nucleotide or sodium salt in the composition, and 5'-adenylic acid and 5' -The weight ratio of uridylic acid is 1:1-1:2.

Further, in the above technical solution, the finished product state of the composition includes powder, granule, tablet, pill, oral liquid, and capsule.

The present invention also provides the use of the above-mentioned composition of uridylic acid, adenylic acid and yeast peptide in the preparation of anti-aging drugs, health products or nutritious foods.

Furthermore, in the above technical solution, the medicines, health products or nutritional foods have the functions of scavenging free radicals in the body, anti-oxidation, good protection and repair of damaged cells, delaying the aging of the body, and restoring youthfulness.

The beneficial effects of the present invention compared with the prior art are as follows:

The invention provides the application of a composition containing three components: 5'-uridylic acid, 5'-adenylic acid, and yeast peptide in anti-aging. First of all, the yeast peptide used in the present invention is a fully nutritional yeast fermentation product. The main component is short peptides of 500-2000Da or less (peptides composed of 5-20 amino acids), and also contains polysaccharides, various amino acids and multiple vitamin minerals. Substances and dietary fiber supplement various nutrients lacking in the aging body. Among them, in the present invention, yeast peptide contains more than 40% of short peptides of 500-2000Da or less (peptides composed of 5-20 amino acids), and contains amino acids that meet the requirements of essential amino acids/total amino acids of more than 40wt.% and branched-chain amino acids/total amino acids. If it is greater than 15wt.%, the effect of the present invention can be achieved. There are no specific restrictions on other nutrients such as polysaccharides, multivitamins, minerals, and dietary fiber, as long as the fermentation products are fermented by fully nutritional yeast using conventional methods. The yeast used in the present invention is preferably Saccharomyces cerevisiae, and the fermentation substrate is preferably molasses, including sugar cane molasses, beet molasses, etc., in which phosphoric acid, zinc sulfate, ammonium sulfate, magnesium sulfate, etc. are added as fermentation nutrients. Research has found that 5'-uridylic acid and 5'-adenylic acid can increase the survival rate of gastrointestinal probiotics, improve the intestinal environment, promote the decomposition of macromolecule proteins, and accelerate the absorption and transportation of short peptides. At the same time, short peptides It can also accelerate the absorption of nucleotides, amino acids, and minerals, and transport them to where they are needed in human tissues. The two promote each other. According to the experimental results, it can be speculated that nucleotides have a regulating effect on the antioxidant effect of amino acids and polypeptides in yeast peptides, so the composition strengthens the antioxidant effect and reduces aging caused by oxidative damage.

Description of the drawings

Figure 1 shows the effect of the composition of uridylic acid, adenylic acid and yeast peptide on glutathione excess in the serum of D-galactose mice according to the present invention. Effect of oxidase (GSH-PX) content. In the figure: ^# indicates that the difference is statistically significant (P<0.05) compared with the model group; ^* indicates that the difference is statistically significant (P<0.05) compared with the blank group.

Figure 2 shows the effect of the composition of uridylic acid, adenylic acid and yeast peptide of the present invention on the superoxide dismutase (SOD) content in the serum of D-galactose mice. In the figure: ^# indicates that the difference is statistically significant (P<0.05) compared with the model group; ^* indicates that the difference is statistically significant (P<0.05) compared with the blank group.

Figure 3 shows the effect of the composition of uridylic acid, adenylic acid and yeast peptide of the present invention on the content of malondialdehyde (MDA) in the serum of D-galactose mice. In the figure: ^# indicates that compared with the model group, the difference is statistically significant (P<0.05).

Figure 4 shows the effect of the composition of uridylic acid, adenylic acid and yeast peptide of the present invention on the lifespan of aging mice.

Detailed ways

The present invention will be further explained below with reference to specific examples. These examples should be understood as illustrative only and not to limit the scope of the present invention in any way. In the following examples, unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used can be purchased from biological or chemical companies.

Example 1

Weigh the raw materials in a clean area according to the weight ratio of 14% 5'-uridylic acid, 14% 5'-adenylic acid, and 72% yeast peptide, and mix evenly with a mixer to obtain a powder. Finished products are sampled and inspected for various indicators in accordance with the quality standards. After passing the inspection, they are stored for future use.

Example 2

Weigh the raw materials in a clean area according to the weight ratio of 41.5% 5'-uridylic acid, 41.5% 5'-adenylic acid, and 17% yeast peptide, and mix evenly with a mixer to obtain a powder. Finished products are sampled and inspected for various indicators in accordance with the quality standards. After passing the inspection, they are stored for future use.

Example 3

Weigh the raw materials in a clean area according to the weight ratio of 49.25% 5'-uridylic acid, 49.25% 5'-adenylic acid, and 1.5% yeast peptide, and mix evenly with a mixer to obtain a powder. Finished products are sampled and inspected for various indicators in accordance with the quality standards. After passing the inspection, they are stored for future use.

Example 4

According to the weight ratio of 18% 5'-uridylic acid, 9% 5'-adenylic acid, and 73% yeast peptide, weigh the raw materials in a clean area and mix them evenly with a mixer to obtain a powder. Finished products are sampled and inspected for various indicators in accordance with the quality standards. After passing the inspection, they are stored for future use.

Example 5

Weigh the raw materials in the clean area according to the weight ratio of 5'-uridylic acid 52%, 5'-adenylic acid 26%, and yeast peptide 22%. Mix evenly with a mixer to obtain powder. Finished products are sampled and inspected for various indicators in accordance with the quality standards. After passing the inspection, they are stored for future use.

Example 6 Antioxidant properties of the composition of uridylic acid, adenylic acid and yeast peptide

The powder prepared in Example 1 was prepared and weighed and then dissolved in a mixed solution of deionized water and ethanol (volume ratio was 1:1) to prepare a mother solution with a concentration of 1 mg/mL, and the DPPH radical scavenging rate was measured. and determination of OH ^- radical scavenging rates.

1. Determination of DPPH free radical scavenging rate:

Dilute the stock solution 100, 50, 25, 10, and 1 times respectively to prepare gradient dilutions with concentrations of 10 μg/mL, 20 μg/mL, 40 μg/mL, 100 μg/mL, and 500 μg/mL.

Take 150 μL of the sample with the above concentration and 150 μL of the DPPH solution with a concentration of 60 mg/L (diluted with absolute ethanol) and place them in a 96-well plate. Mix evenly and store in the dark at 30°C. After 30 minutes, use a microplate reader to measure the absorbance value at 519 nm. The DPPH free radical scavenging rate formula is as follows:

In the formula, A blank is the absorbance of 150 μL deionized water ethanol solution and 150 μL DPPH solution, A sample is the absorbance of 150 μL sample and 150 μL DPPH solution; IC ₅₀ is the concentration of the sample when the DPPH free radical scavenging rate reaches 50%.

2. Determination of OH ^- free radical scavenging rate:

(1) Dilute the mother solution 100, 50, 25, 10, and 1 times respectively to prepare gradient dilutions with concentrations of 10 μg/mL, 20 μg/mL, 40 μg/mL, 100 μg/mL, and 500 μg/mL.

(2) Take 50 μL sample of the above concentration and 50 μL FeSO ₄ solution with a concentration of 6 mmol/L, add 50 μL ethanol-salicylic acid solution with a concentration of 6 mmol/L, place it in a 96-well plate, mix evenly, and add 50 μL concentration It is a 6mmol/L H ₂ O ₂ solution and mix evenly.

(3) After reacting at 37°C for 30 minutes, place the 96-well plate into a microplate reader and measure the absorbance value at 510nm (sample A). The OH ^- free radical scavenging rate formula is as follows:

In the formula, A blank is the absorbance of the sample using DMSO instead of the system, and A sample is the absorbance of the 50 μL sample. IC ₅₀ is the concentration of the sample at which the OH ^- radical scavenging rate reaches 50%.

The powder samples prepared in Examples 2-5 were tested for DPPH free radical scavenging rate and OH ^- free radical scavenging rate according to the above methods. Control group 1: single component 5'-uridylic acid, control group 2: single component 5'-adenylic acid, control group 3: single component yeast peptide, Control group 4: 5'-uridylic acid and 5'-adenylic acid are compounded in a weight ratio of 1:1, control group 5: 5'-guanylic acid 41.5%, 5'-cytidylic acid 41.5%, yeast peptide 17% formulated composition. The experimental data are expressed as mean ± standard deviation, and the experimental results are shown in Table 1.

Table 1 Antioxidant properties of compositions of uridylic acid, adenylic acid and yeast peptides

The results show that compared with the control group 1-5, the compositions of Examples 1-5 all have higher antioxidant properties and show stronger ability to scavenge DPPH free radicals and OH ^- free radicals. Uridylic acid, The antioxidant properties of adenylic acid and yeast peptide are significantly enhanced after compounding. The compositions of Examples 1-5 can more effectively scavenge free radicals in the body, among which Example 2 has the best effect.

The composition prepared by compounding uridylic acid, adenylic acid and yeast peptide in the present invention is compared with the compound compound of single-component uridylic acid, single-component adenylic acid, single-component yeast peptide and two kinds of nucleotides ( 5'-uridylic acid and 5'-adenylic acid compounded at a weight ratio of 1:1) or a mixture of 5'-guanylic acid, 5'-cytidylic acid and yeast peptide all have significant free radical scavenging properties The ability to scavenge DPPH free radicals and scavenge OH ^- free radicals has been doubled. At the same time, it can enhance the activity of antioxidant enzymes in the organism and achieve the purpose of delaying aging.

Example 7 Anti-aging properties of the combination of uridylic acid, adenylic acid and yeast peptide in vitro

Human umbilical vein endothelial cells and PC-12 cells (rat adrenal pheochromocytoma cells) were used as models, and H ₂ O ₂ was used to create the model, and the intracellular ROS levels of the compositions of Examples 1-5 were evaluated. Control group 1: single component 5'-uridylic acid, control group 2: single component 5'-adenylic acid, control group 3: single component yeast peptide, control group 4: 5'-uridylic acid and 5 '-Adenylic acid is compounded at a weight ratio of 1:1, control group 5: a compound compounded of 41.5% 5'-guanylic acid, 41.5% 5'-cytidylic acid, and 17% yeast peptide.

H ₂ O ₂ produces a large amount of ROS after acting on cells, attacking cell membranes, proteins and nucleic acids, causing cell damage. Cells in the logarithmic growth phase were digested into a single cell suspension, adjusted to a cell concentration of 10 ⁵ cells/mL, and seeded in a 96-well cell culture plate, with 100 μL per well and 6 parallel wells in each group. 24 hours after inoculation, the cells grew adherently. The supernatant was discarded, a H ₂ O ₂ medium with a final concentration of 200 μmol/L was added and cultured for 4 hours, and then replaced with the compositions of Examples 1-5 with a final concentration of 50 μg/mL. culture medium Continue culturing for 24 hours. The experiment also set up a blank control without adding H ₂ O ₂ and a model group with only H ₂ O ₂ added. Use an active oxygen detection kit to evaluate the status of intracellular active oxygen: add 10 μmol/L DCFH-DA stock solution to each well ( DMSO dissolved), the final concentration is 1 μmol/L. After continuing to culture for 2 hours, discard the supernatant, add 100 μL of PBS to each well and wash twice. After centrifugation, resuspend the cells in 200 μL of PBS. Use a fluorescent microplate reader to measure the fluorescence of each well under the conditions of excitation wavelength 485 nm and emission wavelength 525 nm. Intensity, calculate the reactive oxygen species content of cells. The experimental data are expressed as mean ± standard deviation, and the experimental results are shown in Table 2.

Table 2 Reactive oxygen species content in human umbilical vein endothelial cells and PC-12 cells

The above results show that compared with the compositions of the model group and the control group 1-5, the compositions of Examples 1-5 can significantly reduce the reactive oxygen species in the model cells. It has good protection and repair effects, among which the effect of Example 2 is the best.

The composition prepared by compounding uridylic acid, adenylic acid and yeast peptide according to the present invention is compared with a single-component uridylic acid, a single-component adenylic acid, a single-component yeast peptide or a combination of two nucleotides ( 5'-uridylic acid and 5'-adenylic acid (compounded at a weight ratio of 1:1) both have the ability to significantly reduce the content of reactive oxygen species in cells damaged by H ₂ O ₂ , and help to reduce aging and oxidatively damaged cells. It can be repaired and its ability to reduce the content of reactive oxygen species in cells has been doubled.

Example 8 Effect of the combination of uridylic acid, adenylic acid and yeast peptide on D-galactose-induced mice

5-week-old BALB/c mice were raised for one week, and 20 of them were intraperitoneally injected with 50 mg/kg D-galactose for 30 days. After 30 days, 10 mice were divided into groups. Experimental group: The compositions of Examples 1-5 were orally administered for 28 days, and D-galactose was injected intraperitoneally at the same time. Control group 1: The same volume of 5'-uridylic acid as the composition was orally administered for 28 days, and D-galactose was injected intraperitoneally at the same time. Control group 2: The same volume of 5'-adenosine as the composition was orally administered for 28 days, and D-galactose was injected intraperitoneally at the same time. Control group 3: The same volume of yeast peptide as the composition was orally administered for 28 days, and D-galactose was injected intraperitoneally at the same time. Control group 4: The same volume of 5'-uridylic acid and 5'-adenylic acid as the composition was administered into the stomach at a weight ratio of 1:1, a total of 28 days, and D-galactose was injected intraperitoneally at the same time. Control group 5: The same volume of 5'-guanylic acid 41.5%, 5'-cytidylic acid 41.5%, and yeast peptide 17% was orally administered for 28 days, and D-half was injected intraperitoneally at the same time. lactose. Model group: intragastric administration of the same volume of PBS buffer as the composition. Blank group: intraperitoneally injected with the same dose of physiological saline as D-galactose, 30 days later, intragastrically administered the same volume of PBS buffer as the composition, and continued intraperitoneal injection for a total of 28 days. On the second day after gastric administration, blood was collected from the orbit and used for subsequent experiments.

The experimental results are shown in Figures 1 to 3. The levels of oxidase SOD and GSH-PX in the serum increased, and malondialdehyde decreased, indicating that the composition of the present invention can alleviate the oxidative damage caused by D-galactose in mice. Compared with the control groups 1-4, the compositions described in Examples 1-5 all showed stronger ability to resist oxidative damage, and uridylic acid and adenylic acid acted synergistically with yeast peptide, and the effect was significant.

Example 9 Delaying effect of the combination of uridylic acid, adenylic acid and yeast peptide on aging in naturally aging mice

Eight-week-old BALB/c mice were randomly divided into two groups, with 10 mice in each group. In the experimental group, the compositions of Examples 1-5 were administered by gavage until the mice died of natural aging. Blank group: The same amount of physiological saline as mentioned above was administered by gavage until the mice aged and died naturally. Control group 1: The same volume of 5’-uridylic acid as the composition was orally administered until the mice died of natural aging. Control group 2: The same volume of 5’-adenosine as the composition was administered by gavage until the mice aged and died naturally. Control group 3: The same volume of yeast peptide as the composition was orally administered until the mice aged and died naturally. Control group 4: The same volume of 5’-uridylic acid and 5’-adenylic acid compounded at a weight ratio of 1:1 was administered to the mice until the mice died of natural aging. Control group 5: A compound containing 41.5% 5'-guanylic acid, 41.5% 5'-cytidylic acid, and 17% yeast peptide in the same volume as the composition was orally administered until the mice aged and died naturally.

The growth and lifespan of mice in each group were calculated separately, and the results are shown in Figure 4. The composition of the present invention can effectively delay the natural aging of mice.

For any person familiar with the art, without departing from the scope of the technical solution of the present invention, they can use the technical content disclosed above to make many possible changes and modifications to the technical solution of the present invention, or modify it into equivalent changes. Example. Therefore, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention should still fall within the protection scope of the technical solution of the present invention.

Claims

A composition of uridylic acid, adenylic acid and yeast peptide, characterized in that the composition includes 5'-uridylic acid, 5'-adenylic acid and yeast peptide, and has anti-aging function.
The composition according to claim 1, characterized in that the composition includes components in the following weight ratio: 5'-uridylic acid 14-63%, 5'-adenylic acid 9-50%, yeast Peptides 1.5-77%.
The composition according to claim 1, characterized in that the composition includes components in the following weight ratio: 5'-uridylic acid 40-60%, 5'-adenylic acid 25-50%, yeast Peptides 10-30%.
The composition according to claim 1, characterized in that the composition includes components in the following weight ratio: 5'-uridylic acid 41.5%, 5'-adenylic acid 41.5%, and yeast peptide 17%; Or 5'-uridylic acid 52%, 5'-adenylic acid 26%, yeast peptide 22%.
The composition according to any one of claims 1 to 4, characterized in that in the composition, 5'-uridylic acid and 5'-adenylic acid are expressed as 5'-monophosphate nucleotides or sodium salts. It exists in the form, and the weight ratio of 5'-adenylic acid and 5'-uridylic acid is 1:1-1:2.
The composition according to any one of claims 1 to 5, characterized in that the finished product state of the composition includes powder, granule, tablet, pill, oral liquid and capsule.
The application of the composition of uridylic acid, adenylic acid and yeast peptide according to any one of claims 1 to 6 in the preparation of anti-aging drugs, health products or nutritional foods.
The application according to claim 7, characterized in that the medicine, health product or nutritional food has the functions of scavenging free radicals in the body, anti-oxidation, good protection and repair of damaged cells, delaying the aging of the body, and restoring youthfulness.