Pentapeptide with HMGCR inhibitory activity

Abstract

The invention discloses a pentapeptide with HMGCR inhibitory activity. The pentapeptide is characterized in that the sequence of the pentapeptide is amino terminal-PGDYP-carboxyl terminal. The peptide has the advantages of good water solubility, no toxicity, no sensitization, good gastrointestinal tract simulation stability and the like, has a remarkable inhibition effect on HMGCR, and can be used as an auxiliary component to develop a medicine or functional food with the characteristic of lowering cholesterol.

Description

Technical Field

[0001] The present invention belongs to the field of peptide biotechnology, and particularly relates to a pentapeptide with HMGCR inhibitory activity. Background Art

[0002] Cardiovascular disease (CVD) has become the most major disease endangering human health and causing human death in today's society. Hypercholesterolemia is one of the most major risk factors leading to the occurrence and development of CVD, which is mainly characterized by the increase in the content of total cholesterol and low-density lipoprotein cholesterol in the blood. 3-Hydroxy-3-methyl glutaryl coenzyme A reductase (HMGCR) is the key rate-limiting enzyme involved in the initial stage of the endogenous cholesterol synthesis reaction. Interrupting the production of mevalonic acid by inhibiting the activity of HMGCR to reduce cholesterol content is still the most popular and mainstream method in the field of cholesterol reduction. Currently, statins (such as pravastatin, simvastatin, atorvastatin, etc.), the most widely used cholesterol-lowering drugs clinically, inhibit the activity of HMGCR competitively and block the endogenous cholesterol synthesis to reduce the cholesterol content in the body. However, studies have found that long-term use of statins will produce toxic and side effects such as muscle weakness, rhabdomyolysis and liver injury. Therefore, developing new active ingredients or functional factors that are safer, non-toxic and side effects and can specifically target HMGCR to improve cholesterol metabolism has become an important direction and effective way to prevent and improve hypercholesterolemia and reduce the risk of the occurrence and development of CVD. Bioactive peptides have attracted wide attention due to their advantages of safety, non-toxic and side effects, high tissue permeability, easy synthesis and modification. Summary of the Invention

[0003] The present invention discloses a pentapeptide with HMGCR inhibitory activity, and its amino acid sequence is: amino terminus - PGDYP - carboxyl terminus, which can be tried to be used as an auxiliary component to develop drugs or functional foods with cholesterol-lowering characteristics. Brief Description of the Drawings

[0004] Figure 1 It is the inhibitory rate of HMGCR activity of the peptide PGDYP of the invention. Detailed Embodiments

[0005] Based on the single-letter sequences of 20 natural amino acids, 400 dipeptides, 8000 tripeptides, 160000 tetrapeptides, and 3200000 pentapeptides were exhaustively constructed using a Python program to obtain the peptide sequence names. Based on the 3368400 peptide sequences constructed above, various peptide properties were predicted (including biological activity, water solubility, toxicity, allergenicity, and simulated gastrointestinal stability). Further, peptides with resistance to simulated gastrointestinal enzymatic digestion, good water solubility, non-toxicity, high biological activity, and non-allergenicity were used for HMGCR target fishing, and peptides containing the HMGCR target with a Probability value ranking in the top five among the target rankings were screened out. Further, molecular docking was performed to evaluate the docking energy value, and molecular dynamics simulation was used to calculate the binding free energy to screen for the peptide segment that binds most tightly to HMGCR. The HMGCR inhibitory activity was measured using an HMGCR detection kit, and the specific operation steps are as follows:

[0006] 1) Blank group: 184 μL of 1× assay buffer, 4 μL of NADPH, and 12 μL of HMG-CoA. 2) Control group: 182 μL of 1× assay buffer, 4 μL of NADPH, 12 μL of HMG-CoA, and 2 μL of HMGCR. 3) Experimental group: 181 μL of 1× assay buffer, 1 μL of the sample, 4 μL of NADPH, 12 μL of HMG-CoA, and 2 μL of HMGCR. At 37 °C and 340 nm, the absorbance value was measured every 20 s for 10 min. The calculation formula for the HMGCR activity inhibition rate is as follows:

[0007]

[0008] Calculated that the IC 50 value of the peptide of the present invention for HMGCR inhibition is 82.95 μM, and its inhibitory activity is significantly stronger than that of multiple reported HMGCR inhibitory peptides, indicating that the peptide of the present invention has good HMGCR inhibitory activity, and its sequence is amino-terminal -PGDYP- carboxyl-terminal.

[0009] In actual production, the peptide of the present invention can be prepared by the Fmoc solid-phase polypeptide chemical synthesis method and added to health products or drugs in the form of granules, powders, or liquids dissolved in water.

[0010] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. A pentapeptide having HMGCR inhibitory activity, characterized in that Its sequence is: amino terminus-PGDYP-carboxyl terminus.

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