CN-122011107-A - Polypeptide, preparation method thereof and application thereof in delaying muscle attenuation

CN122011107ACN 122011107 ACN122011107 ACN 122011107ACN-122011107-A

Abstract

The invention discloses a polypeptide, a preparation method and application thereof in delaying muscle attenuation, and the amino acid sequence is Ile-Arg-Leu-Ser-Phe-Asn-Pro-Thr. The polypeptide of the amino acid sequence can effectively relieve the aging-related markers, improve lipid metabolism abnormality, promote muscle function, improve muscle attenuation, has the advantages of safety, no side effect and the like, and can be applied to products for improving muscle attenuation and related symptoms thereof.

Inventors

REN JIAOYAN
MI LIJUAN
XU YONGZHAO
PANG JINZHU
ZHAO ZIKUAN
ZHOU QIMENG

Assignees

内蒙古蒙牛乳业（集团）股份有限公司

Dates

Publication Date: 20260512
Application Date: 20260409

Claims (10)

1. A polypeptide is characterized in that the amino acid sequence is shown as SEQ ID No. 1, and is Ile-Arg-Leu-Ser-Phe-Asn-Pro-Thr; Wherein Ile is the amino acid corresponding residue of isoleucine, arg is the amino acid corresponding residue of arginine, leu is the amino acid corresponding residue of leucine, ser is the amino acid corresponding residue of serine, phe is the amino acid corresponding residue of phenylalanine, asn is the amino acid corresponding residue of asparagine, pro is the amino acid corresponding residue of proline, and Thr is the amino acid corresponding residue of threonine.
2. The method for producing a polypeptide according to claim 1, wherein the polypeptide is produced by a chemical solid-phase synthesis method or an enzymatic method.
3. The method according to claim 2, wherein the chemical solid phase synthesis method comprises coupling corresponding amino acids one by one on a solid phase carrier to obtain peptide resin, and performing cleavage to obtain target polypeptide; The enzymolysis method comprises the steps of mixing pretreated milk protein-containing raw materials with distilled water, regulating pH, heating to an enzymolysis temperature, respectively adding a first protease and a second protease for enzymolysis, inactivating enzymes after enzymolysis to obtain an enzymolysis solution, centrifuging the enzymolysis solution, and passing through a membrane to obtain a preliminary enzymolysis product, wherein the preliminary enzymolysis product contains polypeptides with amino acid sequences shown in SEQ ID No. 1.
4. The method according to claim 3, wherein the solid phase carrier is resin, and the peptide resin is obtained by respectively obtaining a solid phase carrier preloaded with a first amino acid and a protective amino acid, and coupling the protective amino acid and the resin one by one according to the amino acid sequence from the C end to the N end of the polypeptide; In the enzymolysis process, the distilled water is mixed with the milk protein in the milk protein-containing raw material according to the proportion of (10-20), the pH is regulated to 6.0-8.5, the enzymolysis temperature is 52-58 ℃, the total enzyme bottom ratio is 1 (20-100) in the enzymolysis process, the total enzyme bottom ratio is 4-6 hours, the first protease comprises one or more of serine protease, plant protease, animal protease, bacillus licheniformis protease and thermophilic bacillus protease, and the second protease comprises one or more of bacillus subtilis protease, bacillus licheniformis protease, plant protease, animal protease, alkaline protease, streptomycete protease and black mold protease fermentation protease.
5. The method according to claim 4, wherein the protected amino acid is a single amino acid N-terminal to the Fmoc-protected amino acid, and comprises Fmoc-Pro-OH, fmoc-Asn (Trt) -OH, fmoc-Phe-OH, fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Arg (Pbf) -OH and Fmoc-Ile-OH; The solid phase carrier preloaded with the first amino acid is Fmoc-Thr (tBu) -WANG RESIN, wherein WANG RESIN is a solid phase carrier Wang resin.
6. The method according to claim 4, wherein the coupling reagent used in the coupling process comprises a condensing agent and a base.
7. The method of claim 6, wherein the condensing agent comprises tetrafluoroboric acid-O- (benzotriazol-1-yl) -N, N' -tetramethylurea; the base comprises N, N-diisopropylethylamine.
8. Use of a polypeptide in the preparation of a product for delaying muscle attenuation, wherein the polypeptide is a polypeptide according to claim 1 or a polypeptide prepared by a method according to any one of claims 2 to 7.
9. The use according to claim 8, wherein the product delays muscle decline by reducing the accumulation of age-related markers, including lipofuscin and lipid droplets, improving lipid metabolism abnormalities, and significantly enhancing motor function of the body.
10. The use according to claim 8, wherein the dosage form of the product comprises solutions, oral liquids, capsules, microcapsule powders, tablets, granules, emulsions.

Description

Polypeptide, preparation method thereof and application thereof in delaying muscle attenuation Technical Field The invention relates to the technical field of active peptides, in particular to a polypeptide, a preparation method thereof and application thereof in delaying muscle attenuation. Background Muscle wasting syndrome, abbreviated sarcopenia, refers to a progressive and systemic loss of muscle mass, strength or function associated with aging, with disease code ICD-10-CM (M62.84). Skeletal muscle is an important tissue in vertebrates, plays an important role in metabolism, daily physical activity, and maintaining posture and balance, and muscle attenuation of skeletal muscle causes a series of health problems such as fall, functional decline, dyslipidemia, chronic diseases, weakness, death, and the like. Muscle wasting syndrome increases with age. Currently, there are no approved drugs on the market for the treatment of muscle weakness, and the accepted intervention efforts are resistance training and nutritional supplements. The decrease in muscle protein mass is a result of an imbalance between protein synthesis and protein breakdown. Protein synthesis requires the availability of sufficient amino acids from the diet, with daily recommended protein intake of 1.0 to 1.2 g/kg body weight for elderly sarcopenia patients over 65 years of age, rather than 0.8 g/kg body weight for standard adults, but meeting higher dietary protein requirements is challenging. At the same time, the effective digestion and absorption of protein is a necessary condition for maintaining the synthesis of muscle protein, and the ubiquitous digestion and function decline of the old further limit the actual utilization rate of the protein. Bioactive peptides are specific amino acid sequences derived from proteins, and exhibit different biological activities due to the influence of factors such as molecular weight, amino acid composition and sequence, net charge, hydrophobicity, and the like. A large number of researches indicate that the polypeptide has various nutrition and health care functions. Currently, patent document CN119060126a discloses an oligopeptide with a function of delaying muscle attenuation, which comprises tetrapeptides with amino acid sequences of LWPF and/or LPGF, tripeptides with amino acid sequences of FEP, and the like, and the oligopeptides have the effect of improving muscle attenuation, but have limited effect through experiments. Disclosure of Invention Therefore, the invention aims to provide a polypeptide capable of better delaying muscle attenuation, a preparation method thereof and application thereof in delaying muscle attenuation. A polypeptide with an amino acid sequence shown as SEQ ID No. 1 is Ile-Arg-Leu-Ser-Phe-Asn-Pro-Thr, IRLSFNPT for short; Wherein Ile is the amino acid corresponding residue of isoleucine, arg is the amino acid corresponding residue of arginine, leu is the amino acid corresponding residue of leucine, ser is the amino acid corresponding residue of serine, phe is the amino acid corresponding residue of phenylalanine, asn is the amino acid corresponding residue of asparagine, pro is the amino acid corresponding residue of proline, and Thr is the amino acid corresponding residue of threonine. The polypeptide of the amino acid sequence is octapeptide, the molecular weight is 947.12 Da, and the chemical formula is shown as the formula I; formula I: 。 the preparation method of the polypeptide adopts a chemical solid-phase synthesis method or an enzymolysis method to prepare the polypeptide. Further, the chemical solid-phase synthesis method comprises coupling corresponding amino acids one by one on a solid-phase carrier according to the amino acid sequence shown in SEQ ID NO. 1 to obtain peptide resin, and splitting to obtain the target polypeptide. Furthermore, the solid phase carrier is resin, and the peptide resin is obtained by respectively obtaining the solid phase carrier preloaded with the first amino acid and the protective amino acid, and coupling the protective amino acid and the resin one by one according to the amino acid sequence from the C end to the N end of the polypeptide. The protective amino acid is a single amino acid adopting Fmoc protective amino acid N end, and comprises Fmoc-Pro-OH, fmoc-Asn (Trt) -OH, fmoc-Phe-OH, fmoc-Ser (tBu) -OH, fmoc-Leu-OH, fmoc-Arg (Pbf) -OH and Fmoc-Ile-OH; The solid phase carrier preloaded with the first amino acid is Fmoc-Thr (tBu) -WANG RESIN, wherein WANG RESIN is a solid phase carrier Wang resin. Specifically, when the polypeptide is synthesized by a solid-phase synthesis method, a standard Fmoc scheme is adopted, a solid-phase carrier is used as a resin, wang resin (CAS: 1365700-43-1) preloaded with first amino acid is used as the resin, namely Fmoc-Thr (tBu) -WANG RESIN, fmoc protected amino acid N-ends are adopted, each protected amino acid is Fmoc-Pro-OH, fmoc-Asn (Trt) -OH, fmoc-Phe-OH, fmoc-Ser (tBu) -OH, fmoc-Leu-OH