CN114652850B

CN114652850B - Polypeptide coupled NMN compound with anti-aging effect and application thereof

Info

Publication number: CN114652850B
Application number: CN202210329513.1A
Authority: CN
Inventors: 戴政清; 张凌云; 王宇恩; 马亚平
Original assignee: Shenzhen Shenchuang Biopharmaceutical Co ltd
Current assignee: Shenzhen Shenchuang Biopharmaceutical Co ltd
Priority date: 2022-03-31
Filing date: 2022-03-31
Publication date: 2023-12-29
Anticipated expiration: 2042-03-31
Also published as: CN114652850A

Abstract

A polypeptide coupling NMN compound with anti-aging effect and application thereof belong to the technical field of biological medicine. The structural general formula of the polypeptide coupled NMN compound is X1-NMN, wherein X1 represents a polypeptide sequence, and NMN is beta-nicotinamide mononucleotide. Polypeptide conjugated NMN compounds are linked by the formation of an ester bond between a hydroxyl group on NMN and a carboxyl group on the polypeptide sequence. According to the invention, through coupling of the polypeptide and NMN, the functional polypeptide carries NMN to permeate the skin surface layer to release NMN molecules, so that the defect that NMN cannot be absorbed through skin is overcome, and through the effect of NMN on skin, the content of NMN in skin tissues can be greatly increased, thereby enhancing the skin resistance, improving the skin immunity, protecting the skin from environmental invasion, inhibiting skin aging, and having obvious anti-wrinkling effect and anti-aging effect.

Description

Polypeptide coupled NMN compound with anti-aging effect and application thereof

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to polypeptide coupling NMN compounds with an anti-aging effect, and application of the compounds.

Technical Field

Representative theories concerning the aging mechanism include the free radical theory, mitochondrial DNA damage theory, and the like. Among them, the free radical theory is one of the representative aging theory, which considers that free radicals are generated in the body at any time, but an effective free radical scavenging system (such as superoxide dismutase) exists at the same time, so that the free radicals in the body are maintained at normal level. Many substances with oxidation resistance in organisms belong to peptides, and the peptides have excellent emulsifying property and can play a role in mediating an oil-water interface, so that the peptides have important significance in removing excessive free radicals in the body and inhibiting peroxidation of membrane lipid.

Meanwhile, the aging process of human beings is always accompanied with disturbance of immune function, and improvement of immune function is one of the most effective methods for resisting aging. The polypeptide is the most obvious way to regulate the immune function as the most basic substance for maintaining the cell activity and supplying the cell energy. The polypeptide can delay the aging of specific immune organs and immune cells, delay the aging of nonspecific immune tissues and immune cells, repair damaged specific immune organs and immune cells and activate the immune function of human body.

NMN, β -nicotinamide mononucleotide, is an inherent substance in the human body and is also enriched in some fruits and vegetables, but is present in very small amounts. In humans, NMN is converted to nad+ (nojia factor), which is involved in intracellular material, energy metabolism, providing 95% of the energy required by the human body. NAD+ exerts a fundamental influence on human health, but as the content of NAD+ in a human body gradually decreases with age, communication between mitochondria and nuclei is impaired, and the reduction of NAD+ also impairs the ability of cells to produce energy, resulting in damage to DNA replication, inability of cells to divide and operate normally, and further transformation into senescent cells. To a certain extent, NMN can delay the aging of human bodies, especially in skin tissues, NMN can slow down and promote the metabolism of keratinocytes to a certain extent, accelerate the decomposition of melanin and make the skin more white and clear. However, NMN itself has poor absorption rate at the skin surface layer, and it is difficult to obtain a satisfactory effect by direct application.

At present, in terms of anti-aging, human beings mainly comprise anti-aging drugs and functional polypeptide anti-aging cosmetics, such as vitamin E, aspirin, metformin and the like. However, these drugs have side effects on organs of the human body such as the stomach for a long period of time, and drug resistance also affects the effect of the drug use. Other soybean polypeptides, carnosine, and short peptides such as argireline are also commonly used in cosmetics, mainly for anti-wrinkle and anti-aging of skin. The products of these polypeptides are of various varieties, but the specific mechanism of action is not clear and the anti-aging effect is not very effective. The invention patent with publication number of CN 111419784A discloses an anti-wrinkle eye cream containing NMN, and a preparation method and application thereof. However, the formulation is complex and the formation of a skin care product is not a synthetic new anti-aging compound, which is essentially different from the present invention.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to design and provide a polypeptide compound with an anti-aging effect and application thereof, has good anti-aging and anti-wrinkling effects, and has great development prospects in medicines and cosmetics.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

a polypeptide coupled NMN compound with an anti-aging effect is characterized in that the structural general formula of the polypeptide coupled NMN compound is X1-NMN, wherein X1 represents a polypeptide sequence, and NMN is beta-nicotinamide mononucleotide.

The polypeptide coupling NMN compound with the anti-aging effect is characterized in that the polypeptide coupling NMN compound is connected through an ester bond formed by a hydroxyl group on NMN and a carboxyl group on a polypeptide sequence, and the structure of the polypeptide coupling NMN compound is shown as a formula I or a formula II:

the polypeptide coupling NMN compound with the anti-aging effect is characterized in that the polypeptide sequence X1 is acetyl tetrapeptide-11, and the amino acid sequence of the acetyl tetrapeptide-11 is Ac-Pro-Pro-Tyr-Leu-OH.

The polypeptide sequence X1 is palmitoyl hexapeptide-12, and the amino acid sequence of palmitoyl hexapeptide-12 is Pal-Val-Gly-Val-Ala-Pro-Gly-OH.

The polypeptide sequence X1 is palmitoyl tripeptide-1, and the amino acid sequence of the palmitoyl tripeptide-1 is Pal-Gly-His-Lys-OH.

The polypeptide sequence X1 is palmitoyl pentapeptide-4, and the amino acid sequence of palmitoyl pentapeptide-4 is Pal-Lys-Thr-Thr-Lys-Ser-OH.

The polypeptide sequence X1 is tetrapeptide-3, and the amino acid sequence of the tetrapeptide-3 is H-Lys-Gly-His-Lys-OH.

The polypeptide sequence X1 is carnosine, and the amino acid sequence of the carnosine is H-beta-Ala-His-OH.

The polypeptide sequence X1 is argireline, and the amino acid sequence of the argireline is Ac-Glu ¹ -Glu-Met-Gln-Arg-Arg-NH ₂ 。

The polypeptide coupled NMN compound with the anti-aging effect is characterized in that Glu on the argireline ¹ The side chain carboxyl is connected with the hydroxyl on NMN through ester bond.

The application of any polypeptide coupled NMN compound or pharmaceutically acceptable salt thereof in preparing anti-aging drugs.

The application is characterized in that the anti-aging medicine preparation comprises injection, freeze-dried powder injection, capsule, tablet, granule, paste, spray and oral liquid.

An anti-aging agent comprising any one of the polypeptide-conjugated NMN compounds or a pharmaceutically acceptable salt thereof.

The application of any polypeptide coupled NMN compound or pharmaceutically acceptable salt thereof in preparing anti-aging or anti-wrinkling cosmetics.

Compared with the prior art, the invention has the following beneficial effects:

1. according to the invention, through coupling of the polypeptide and NMN, a novel compound is obtained, NMN molecules are introduced while the polypeptide has an anti-aging function, and the NMN can be converted into NAD+ (norganin) in a human body, so that the novel compound has an obvious anti-aging function.

2. According to the invention, through coupling of the polypeptide and NMN, the functional polypeptide carries NMN to permeate the skin surface layer to release NMN molecules, so that the defect that NMN cannot be absorbed through skin is overcome, and through the effect of NMN on skin, the content of NMN in skin tissues can be greatly increased, thereby enhancing the skin resistance, improving the skin immunity, protecting the skin from environmental invasion, inhibiting skin aging, and having obvious anti-wrinkling effect and anti-aging effect.

Drawings

FIG. 1 is a HPLC analysis chart of crude compound 1 in example 1;

FIG. 2 is a MS analysis chart of peak 1 of the crude HPLC analysis result of compound 1 in example 1;

FIG. 3 is a MS analysis chart of peak 2 of the crude HPLC analysis result of compound 1 in example 1;

FIG. 4 is a molecular structure diagram of peak 1 in example 1;

FIG. 5 is a molecular structure diagram of peak 1 in example 1;

FIG. 6 is a molecular structure diagram of two molecular structure compounds 2-1 and 2-2 of carnosine-NMN compound in example 2;

FIG. 7 is a molecular structure diagram of two molecular structure compounds 3-1 and 3-2 of the acetyltetrapeptide-11-NMN polypeptide compound of example 3.

Detailed Description

The invention will be further described with reference to examples and figures.

Example 1: synthesis of Alorelin-NMN polypeptide compounds

Using Fmoc solid-phase polypeptide synthesis technique, sieberAmide Resin (sub=0.6 mmol/g) was selected as starting resin, and according to standard coupling method for Fmoc solid-phase polypeptide synthesis (3 times of amino acid addition, coupling system was HOBt/DIC), fmoc-Arg (Pbf) -OH, fmoc-gin (Trt) -OH, fmoc-Met-OH, fmoc-Glu (OtBu) -OH, fmoc-Glu (OAll) -OH were sequentially coupled according to the peptide sequence of argireline, and after the coupling was completed, acetylation was completed on the resin with acetic anhydride/pyridine (1:1, 30 times resin equivalent) to obtain Ac-Glu (OAll) -Met-gin (Pbf) -Arg (Pbf) -SieberAmide Resin.

For the above peptide resin, phenylsilane (40 times resin equivalent) and Pd were used ⁰ (Ph ³ P) ₄ (0.1 times resin equivalent), the OAll protecting group was removed by reaction in DCM for 2 hours to give Ac-Glu-Glu (OtBu) -Met-Gln (Trt) -Arg (Pbf) -Arg (Pbf) -SieberAmide Resin.

The above peptide resin with OAll protecting groups removed was washed with DCM containing 1% tfa for 30 minutes, suction filtered and the filtrate was collected. This step completes cleavage of the fully protected peptide from the resin to give Ac-Glu-Glu (OtBu) -Met-Gln (Trt) -Arg (Pbf) -Arg (Pbf) -NH ₂ 。

The fully protected peptide fragment obtained above was added to THF, HOSu (1.2 eq), DCC (1.2 eq) was added under ice water bath, reacted for 8 hours at room temperature, DCM was added, the solution was washed 3 times with saturated sodium bicarbonate, purified water was 2 times, the organic phase was dried over anhydrous sodium carbonate, and spin-dried to give fully protected active OSu ester: ac-Glu (OSu) -Glu (OtBu) -Met-Gln (Trt) -Arg (Pbf) -Arg (Pbf) -NH ₂ 。

The fully protected active OSu ester described above was dissolved in DCM, DMAP (0.2 eq) was added, DIEA (1.5 eq) was added, then NMN (1.0 eq) was added dropwise and reacted for 8 hours. Adding acetic acid to neutralize the reaction solution to be neutral, and then spin-drying the reaction solution to obtain oily matter for later use.

TFA was added to the oil: TIS: EDT: phOH: h ₂ O=90:3:3:2:2 (volume ratio) lysate, reacted at room temperature for 1.5 hours, then poured into anhydrous diethyl ether to precipitate a white solid, centrifuged, the solid washed with anhydrous diethyl ether, and dried in vacuo to give crude polypeptide coupled NMN as a white solid: ac-Glu (NMN) -Glu-Met-Gln-Arg-Arg-NH ₂ (molecular weight: 1223.2), labeled compound 1.

HPLC analysis was performed on Compound 1, and the results are shown in FIG. 1. Considering that two bare hydroxyl groups of NMN do not react selectively during the reaction, and that fig. 1 shows two main peaks, both peak 1 and peak 2 in fig. 1 are analyzed by mass, and the results show that both are target molecular weights, see fig. 2 and 3 in particular. For the above peaks 1 and 2, we purified both peaks by HPLC to obtain pure peak 1 and pure peak 2 with purities greater than 99%. The structure of the pure product of peak 1 and the pure product of peak 2 were confirmed by hydrogen spectrum and carbon spectrum analysis of nuclear magnetic resonance, and the result shows that the molecular structure of peak 1 (we marked as compound 1-1) is shown in fig. 4, and the molecular structure of peak 2 (we marked as compound 1-2) is shown in fig. 5.

Example 2: synthesis of carnosine-NMN polypeptide compounds

2-CTC Resin (sub=0.8 mmol/g) is selected as a starting Resin by using Fmoc solid-phase polypeptide synthesis technology, fmoc-His (Trt) -OH and Boc-beta-Ala-OH are sequentially coupled according to the peptide sequence of carnosine according to a standard coupling method (amino acid feeding multiple is 3 times, a coupling system is HOBt/DIC) of Fmoc solid-phase polypeptide synthesis, and Boc-beta-Ala-His (Trt) -2-CTC Resin is obtained after the coupling is completed.

The cleavage of the fully protected peptide fragment from the resin was completed by the addition of DCM containing 1% TFA using the fully protected peptide resin described above in the same manner as in example 1 to give Boc-beta-Ala-His (Trt) -OH.

Then, for the above-mentioned full-protection peptide fragment, we used the same procedure as in the following example 1 to obtain two carnosine-NMN compounds of different structures after HPLC purification, and compound 2-1 and compound 2-2 with purity of more than 99% were obtained, the structures of which are shown in FIG. 6.

Example 3; synthesis of acetyl tetrapeptide-11-NMN polypeptide compounds

Using the same method as in example 2, compound 3-1 and Compound 3-2 having purities of more than 99% were obtained, the structures of which are shown in FIG. 7.

Example 4; influence of polypeptide Compounds on the longevity of caenorhabditis elegans

Culturing caenorhabditis elegans: reference (research on anti-aging of plant extracts by means of model organism caenorhabditis elegans) (Lv Ting, [ academic paper ], major, 2014) the culture method of caenorhabditis elegans was completed to obtain caenorhabditis elegans in stage L4.

Life test of caenorhabditis elegans: to the resulting L4-stage caenorhabditis elegans, 100. Mu.g/ml of 5-FUDR was added to suppress the oviposition of caenorhabditis elegans. Then the caenorhabditis elegans develop into adults, 60 caenorhabditis elegans with good activity are placed in each culture dish, and then the administration is started. Administration included control, 10nM of compound 1-1, compound 1-2, compound 2-1, compound 2-2, compound 3-1, compound 3-2 (all obtained by example 1-3). The first day of administration was recorded as the first day, followed by daily administration, and the number of caenorhabditis elegans surviving and dying was observed daily until the last caenorhabditis elegans dying. The basis for judging the death of caenorhabditis elegans is as follows: the caenorhabditis elegans does not react to strong light and knocking flat plates, the pharyngeal muscles under the high-power mirror do not act, finally, the caenorhabditis elegans collides with the head of the caenorhabditis elegans by using the insect picking needle, and the caenorhabditis elegans is still not reacted at any time, so that the caenorhabditis elegans is judged to have died. The dead caenorhabditis elegans will pick up the dishes in time and the experimental results are shown in Table 1.

Table 1: life test experimental result of caenorhabditis elegans

From the statistics in the table above, caenorhabditis elegans death occurred on days later in the 6 groups than in the control group, except that the 3-1 group was probably one due to unexpected death on day 2. Meanwhile, the control group had a maximum survival time of 28 days, while the polypeptide compound group had a shortest survival time of 32 days and a longest survival time of 35 days. The above results demonstrate that all 6 polypeptide compounds have a significant effect in prolonging the life of caenorhabditis elegans. The application of the compounds in the fields of anti-aging medicines and functional cosmetics has wide research prospect.

Example 5: preparation method of polypeptide compound oral liquid

Weighing 1g of the 6 polypeptide compounds with the purity of more than 99.0% obtained in the examples 1-3, adding water for injection for dissolution, then adding 0.05g of sucralose, adding 0.02g of lemon flavor essence, adding purified solution to prepare 100ml, mixing uniformly, packaging, and carrying out damp-heat sterilization to obtain the corresponding polypeptide oral liquid.

Example 6: toxicity test of polypeptide compound oral liquid

The 6 polypeptide oral liquids prepared in example 5 were subjected to an acute toxicity test, a genotoxicity test, an Ames test, a mouse sperm abnormality test, a mouse testicle chromosome aberration test, and a mouse feeding test for 30 days according to the relevant requirements of food safety toxicology evaluation procedure. Aiming at the test, according to the related guiding principle and general test methods, the prepared 6 oral liquids are subjected to corresponding tests, and the specific test results are as follows:

acute toxicity test: after 6 oral solutions are administered, the administration is continuously observed for one week, and 120 mice have no obvious adverse reaction or death.

The tolerance of the 6 oral liquids to mice lavage is shown in Table 2:

table 2: lavage tolerance statistics table of 6 oral liquids for mice

Oral liquid	Gastric lavage tolerance (g/kg)	Equivalent to clinical multiple
			Compound 1-1 oral liquid	210	2100
Compound 1-2 oral liquid	210	2100
			Compound 2-1 oral liquid	180	1800
Compound 2-2 oral liquid	170	1700
			Compound 3-1 oral liquid	200	2000
Compound 3-2 oral liquid	190	1900

From the above analysis, all the compound oral liquids of the present invention belong to non-toxic substances according to the acute toxicity setting standard.

Ames et al test results: through experimental research of a general method, the results of the compound oral liquid Ames test, the mouse sperm malformation test and the mouse testicle chromosome aberration test are all negative, which indicates that the compound oral liquid Ames test has no genotoxicity.

Rats were fed for 30 days test results: the compound oral liquid of the invention with different dosages carries out group feeding tests on rats, and the growth and development of animals in experimental groups are good in the test period, and indexes such as weight, ingestion, blood convention, blood biochemistry, organ coefficients and the like are all in a normal range, so that no abnormality is seen in pathological histology examination, and the product has no toxicity.

Long-term toxicity test results: the compound oral liquid of the invention with different dosages is continuously infused into the stomach of a rat for 3 months, animals of an experimental group have no adverse reaction during the test period, all inspection indexes are in a normal range, and main organs of pathological inspection have no toxic pathological change, so that the compound oral liquid has no long-term toxicity.

Example 7: anti-aging test of polypeptide compound oral liquid

The method comprises the following steps: 70 old mice are taken and randomly divided into 7 groups, 10 mice in each group are respectively infused with 60ml/kg of the oral liquid of the invention, and the control group is infused with 0.5 ml/mouse of physiological saline. Under the same conditions, the feed is continuously fed for 30 days. The oral liquid medicine composition of the invention is administered for 2 hours for orbital blood sampling, and is used for measuring SOD and LPO, and the content of hydroxyproline is measured by taking the tail ciliary after the mice are sacrificed. The test results are shown in Table 3.

Table 3: influence of 6 oral liquids on aged mice SOD, LPO, HO-pro (X.+ -.30)

Oral liquid	Number of mice	SOD(ug/gHb)	LPO(mol/ml)	Ho-pro(mg/5mg)
					Physiological saline	10	1298±327	7.13±1.82	3.55±0.28
Compound 1-1 oral liquid	10	2307±389**	4.58±1.33	4.62±0.33
					Compound 1-2 oral liquid	10	2287±403**	4.82±1.25	4.57±0.31
Compound 2-1 oral liquid	10	2458±470**	4.21±1.18	5.03±0.38
					Compound 2-2 oral liquid	10	2386±467**	4.42±1.09	4.94±0.32
Compound 3-1 oral liquid	10	2109±355**	4.80±1.32	4.39±0.26
					Compound 3-2 oral liquid	10	2066±360**	4.91±1.25	4.21±0.25

Note that: p is less than 0.001 compared to saline.

Conclusion of the test:

from the test results, the 6 polypeptide compound oral liquids can improve SOD activity in red blood cells of old mice, reduce the content of LPO in serum, and increase HO-PRO content of rat tails.

SOD is an important antioxidant enzyme in animals and can remove toxic action of free radicals on organisms. The polypeptide compound oral liquid can reduce the generation of LPO in vivo by increasing SOD activity, and remove the damage of free radicals in vivo to the body, thereby reducing the incidence of diseases and delaying aging.

HO-pro is correlated with the presence of aging in an animal, and the aging status of an animal can be detected by measuring HO-pro. Older people and older animals gradually decrease in protein solubility with age, and thus decrease in HO-pro content. The polypeptide compound oral liquid can increase HO-pro content of rat tail ciliary of a rat, so that aging of animals can be inhibited.

Example 8: preparation of polypeptide compound essence

The formulation of the polypeptide compound concentrate is shown in table 4 below:

table 4: polypeptide compound essence formula table

The ingredients of the active substances and the corresponding concentrate numbers in table 4 are shown in table 5:

table 5: active substance components in essence formula and corresponding numbers

Active substances	Essence liquid
		Blank space	Essence-1
NMN	Essence-2
		Compounds 1-1	Essence-3
Compounds 1-2	Essence-4
		Compound 2-1	Essence-5
Compound 2-2	Essence-6
		Compound 3-1	Essence-7
Compound 3-2	Essence-8

The preparation process comprises the following steps: according to the formula shown in Table 4, all components B are added into an emulsifying pot, heated to 80 ℃, kept at the temperature and stirred for 15 minutes, homogenized for 10 minutes, stirred and cooled to 40 ℃, the components A are added in sequence according to the sequence shown in the table, stirred and cooled to 35 ℃, and filtered and discharged to obtain the corresponding essence.

Example 9: experiment of Effect on collagen formation

The effect of polypeptide compounds on collagen formation was determined by measuring the hydroxyproline content in the skin by means of a hydroxyproline kit.

The experimental method comprises the following steps: nude mice that were 8 weeks old (about 20 g) were randomly selected and divided into 8 groups of 10 animals each. Then, the essence-1, the essence-2, the essence-3, the essence-4, the essence-5, the essence-6, the essence-7 and the essence-8 prepared in the example 8 are respectively smeared on the backs of the right ears of each group of nude mice, and each time is 1mL each time. All nude mice were then sacrificed after 30 days according to daily feeding conditions and the skin of the drug site was taken as test tissue.

Weighing 50mg of skin tissues for each test, placing the skin tissues into a corresponding sample test tube, adding a hydrolysis solution in a hydroxyproline determination kit, and hydrolyzing in an oven at 110 ℃ for 30 minutes. After cooling, 1 drop of indicator in the kit is added and shaken well. Then 1mL of essence in the kit is added and shaken well. Then, the pH-adjusted ethyl acetate was added dropwise to each sample tube until the solution in each tube turned yellow-green. 10mL of distilled water was added, 5mL of the mixture was mixed uniformly, 50mg of activated carbon was added, the mixture was centrifuged at a low speed for 10 minutes, and 1mL of the supernatant was obtained.

In addition, a standard solution was added to the test tube as a standard tube. After adding the reagent 1 to each tube and then standing for 5 minutes, adding the reagent 2 and standing for 5 minutes, taking the supernatant of each tube, and measuring the absorbance value under the condition of 550nm wavelength.

Analysis of experimental results: hydroxyproline (HYP) is one of the main components of collagen in the body, so HYP content is an important index reflecting the degree of collagen tissue metabolism and fibrosis. In the experiment, spectrophotometry is adopted to measure the HYP content, and then the content of the HYP with 14% of collagen is used as a standard according to the average data of detection, and converted into the content of the collagen in the skin of the mice. The specific results are shown in Table 6.

Table 6: data sheet of influence of polypeptide compound on skin collagen generation of nude mice

Grouping	Skin collagen content (%), for 30 days
		Essence-1	12.06
Essence-2	16.17
		Essence-3	23.51
Essence-4	21.42
		Essence-5	22.09
Essence-6	20.23
		Essence-7	23.16
Essence-8	20.72

As can be seen from the data in table 6, the collagen content of skin tissue was significantly increased after 30 days of continuous administration of the essence-3, essence-4, essence-5, essence-6, essence-7, essence-8, which were prepared from the polypeptide compounds. Whereas the NMN formulated concentrate-2 had a relatively higher collagen content than the blank concentrate-1, but was significantly inferior to the concentrate with the addition of the polypeptide compound. This demonstrates that the polypeptide compound has a good effect in promoting the production of skin collagen. An increase in collagen will inhibit skin aging.

The foregoing description is only illustrative of the present invention and is not intended to limit the scope of the invention, and any modifications made by the teachings of the present invention, or used directly or indirectly in the related art, are intended to be included within the scope of the present invention.

Claims

1. A polypeptide coupling NMN compound with anti-aging effect or pharmaceutically acceptable salt thereof is characterized in that the structural general formula of the polypeptide coupling NMN compound is X1-NMN, wherein X1 represents a polypeptide sequence, and NMN is beta-nicotinamide mononucleotide;

the polypeptide sequence X1 is selected from one of acetyl tetrapeptide-11, carnosine or argireline;

the polypeptide coupling NMN compound is connected through an ester bond formed by hydroxyl on NMN and carboxyl on a polypeptide sequence, and the structure of the polypeptide coupling NMN compound is shown as a formula I or a formula II:

2. the polypeptide-conjugated NMN compound or a pharmaceutically acceptable salt thereof with anti-aging effect according to claim 1, wherein the amino acid sequence of the acetyl tetrapeptide-11 is Ac-Pro-Pro-Tyr-Leu-OH, the amino acid sequence of the carnosine is H-beta-Ala-His-OH, and the amino acid sequence of the argireline is Ac-Glu ¹ -Glu-Met-Gln-Arg-Arg-NH ₂ 。

3. The polypeptide-conjugated NMN compound or a pharmaceutically acceptable salt thereof having an anti-aging effect according to claim 1, wherein Glu on the Azithrone ¹ The side chain carboxyl is connected with the hydroxyl on NMN through ester bond.

4. Use of a polypeptide-conjugated NMN compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3 in the manufacture of an anti-aging medicament.

5. The use according to claim 4, wherein the anti-aging agent is selected from one of injection, capsule, tablet, granule, paste, spray, and oral liquid.

6. The use according to claim 5, wherein the injection is a lyophilized powder for injection.

7. An anti-aging agent comprising a polypeptide conjugated NMN compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

8. Use of a polypeptide conjugated NMN compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3 in the manufacture of an anti-ageing or anti-wrinkling cosmetic.