Abstract:

P7 lipopeptide was synthesized through solid-phase peptide synthesis （SPPS） and hydrophobic-support-assisted liquid-phase peptide synthesis （LPPS）. Molecular docking technique was used for docking the product with 3OCG protein. MTT assay and Griess reagent were used to detect the biological toxicity and anti-inflammatory activity of P7 lipopeptide. The synthesized P7 lipopeptide was characterized by mass spectrometry. For SPPS and LPPS methods, the yields were 34.45% and 66.31%, respectively; the material consumptions were 129.12 g and 83.08 g, respectively; the waste liquid volumes were 7.12 L and 3.38 L, respectively; PMI were 30.81 and 10.29, respectively; E factors were 1 700.62 and 419.70, respectively. The results of molecular docking showed that the docking binding energy between P7 lipopeptide and p38 protein kinase （3OCG） was the smallest. P7 lipopeptide formed hydrogen bonds with residues ARGA70, ARGA189, TYRA35, ARGA67, ASNA155, and ASPA168, and interacted with residues by van der Waals forces, π-alkyl groups, and carbon hydrogen bonds. P7 lipopeptide had no biological toxicity within the concentration of 32 μmol/L. Different concentrations of P7 lipopeptide could significantly downregulating the release of NO in LPS-induced RAW264.7 cell inflammatory model. In summary, the LPPS method had higher yield and lower raw material consumption compared with SPPS method, making it suitable for industrial production. The results of molecular docking and cell experiments indicated that the designed P7 lipopeptide had good anti-inflammatory and antiosteoporosis activities.

Key words: polypeptide, LPPS method, SPPS method, molecular docking