Pentapeptide with HMGCR inhibitory activity

Abstract

The invention discloses a peptide fragment with HMGCR inhibitory activity. The peptide fragment is characterized in that the amino acid sequence of the peptide fragment is amino terminal-EFPPF-carboxyl terminal. The peptide has the advantages of good water solubility, no toxicity, no sensitization, good gastrointestinal tract stability and the like. The compound shows a remarkable effect in the aspect of inhibiting HMGCR activity, and can be used as an auxiliary material component for developing a medicine or functional food with a cholesterol lowering effect.

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] Hypercholesterolemia is one of the most important risk factors leading to the occurrence and development of cardiovascular diseases, which is mainly characterized by elevated levels of total cholesterol and low-density lipoprotein cholesterol in the blood. 3-Hydroxy-3-methyl glutaryl coenzyme A reductase (HMGCR) is a key rate-limiting enzyme involved in the initial stage of endogenous cholesterol synthesis reaction. Interrupting the production of mevalonic acid by inhibiting HMGCR activity to reduce cholesterol content is still a relatively common and effective method in the field of cholesterol reduction. Currently, statins (such as pravastatin, simvastatin, atorvastatin, etc.), the most widely used cholesterol-lowering drugs in clinical practice, inhibit HMGCR activity competitively, block endogenous cholesterol synthesis, and thus reduce the cholesterol content in the body. However, studies have found that long-term use of statins may cause side effects such as muscle weakness, rhabdomyolysis, and liver damage. Therefore, developing new active ingredients or functional factors that are safe, non-toxic, can specifically target HMGCR, and improve cholesterol metabolism has become an important direction and effective approach for preventing and improving hypercholesterolemia and reducing the occurrence and development of cardiovascular diseases. Bioactive peptides have attracted extensive attention in recent years due to their advantages such as high safety, non-toxic side effects, good tissue permeability, and easy synthesis and modification, and have become important candidates for the development of new cholesterol-lowering drugs. Summary of the Invention

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

[0004] The attached drawings of the present invention Figure 1 are for the HMGCR activity inhibition rate of the inventive peptide EFPPF. Detailed Description of the Invention

[0005] Based on the single-letter sequences of 20 natural amino acids, 400 dipeptides, 8,000 tripeptides, 160,000 tetrapeptides, and 3,200,000 pentapeptides were exhaustively constructed using a Python program. Based on the 3,368,400 peptide sequences constructed above, various peptide property predictions were carried out, including evaluations of biological activity, water solubility, toxicity, allergenicity, and simulated gastrointestinal stability. Peptides with resistance to simulated gastrointestinal enzymatic hydrolysis, good water solubility, non-toxicity, high biological activity, and non-allergenicity were further screened for HMGCR target fishing, and peptides containing the HMGCR target with a Probability value ranking in the top five among the target rankings were selected. Subsequently, molecular docking technology was used to evaluate the binding energy of each peptide segment to HMGCR, and combined with molecular dynamics simulation calculations and free energy changes, the peptide segment EFPPF that tightly binds to HMGCR was screened out. Finally, the HMGCR inhibitory activity of the screened peptide segments was determined 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, 12 μL of HMG-CoA. 2) Control group: 182 μL of 1× assay buffer, 4 μL of NADPH, 12 μL of HMG-CoA, 2 μL of HMGCR. 3) Experimental group: 181 μL of 1× assay buffer, 1 μL of sample, 4 μL of NADPH, 12 μL of HMG-CoA, 2 μL of HMGCR. At 37 °C and 340 nm, measure the absorbance value every 20 s for 10 min. The calculation formula for the HMGCR activity inhibition rate is as follows:

[0007]

[0008] It was calculated that the IC 50 value of the peptide of the present invention for HMGCR inhibition was 116.44 μM, and its inhibitory activity was 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 - EFPPF - 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 modifications and variations 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 with HMGCR inhibitory activity, characterized in that Its sequence is: amino-terminal - EFPPF - carboxyl-terminal.

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