P7脂肽的合成工艺与生物活性研究

doi:10.3969/j.issn.2097-2806.2024.09.005

摘要/Abstract

摘要：

对P7脂肽的合成工艺及生物活性进行研究。采用固相多肽法（SPPS）和疏水载体辅助液相法（LPPS）人工合成P7脂肽；利用分子对接技术将合成的化合物与受体蛋白（3OCG）进行对接；采用MTT法与Griess试剂法检测P7脂肽的生物毒性和抗炎活性。成功合成P7脂肽并对其进行质谱表征；SPPS法与LPPS法的收率分别为34.45%和66.31%，物料消耗量分别为129.12和83.08 g，废液产生量分别为7.12和3.38 L，PMI分别为30.81，10.29，E-因子分别为1 700.62，419.70；分子对接结果表明，P7脂肽与p38蛋白激酶（3OCG）的对接结合能最小，GHK分子与残基ARG67、LYS53和GLY36形成氢键，P7肽与残基ALAA34、TYRA35、THRA185形成氢键，P7脂肽与残基ARGA70、ARGA189、TYRA35、ARGA67、ASNA155、ASPA168形成氢键并与残基存在范德华力、pi-烷基、C-H键等相互作用力；P7脂肽在32 μmol/L浓度以内无生物毒性；不同浓度的P7脂肽对LPS诱导RAW264.7细胞炎症模型中NO的释放具有很好的下调作用。通过对P7脂肽的合成研究，表明LPPS法的总体收率比SPPS法收率高且物料消耗量及“三废”产生量少，更适合于工业化生产；分子对接与细胞实验结果表明设计合成的P7脂肽具有良好的抗炎抗骨质疏松活性。

关键词: 多肽, LPPS法, SPPS法, 分子对接

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

中图分类号:

TQ658

张文杰, 石峰, 咸瑞卿, 董玉香, 郭凯, 姬胜利. P7脂肽的合成工艺与生物活性研究[J]. 日用化学工业（中英文）, 2024, 54(9): 1050-1058.

Wenjie Zhang, Feng Shi, Ruiqing Xian, Yuxiang Dong, Kai Guo, Shengli Ji. Study on the synthesis and biological activity of P7 lipopeptide[J]. China Surfactant Detergent & Cosmetics, 2024, 54(9): 1050-1058.

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方法	PMI	E-因子	收率/%	物料消耗量/g	AA消耗量/g	m （HOBT） / g	m （DIC） / g	V （废液） /L	V （THF） /L	V （CAN） /L	V （DCM） /L	V （DMF） /L	V （MeOH） /mL
SPPS	30.81	1 700.62	34.45	129.12	79.63	16.31	13.74	7.12	—	—	2.24	3.58	400
LPPS	10.29	419.70	66.31	83.08	47.79	9.72	8.22	3.38	1.74	0.95	0.12	0.06	80

化合物	Docking Score	Confidence Score
GHK	-140.45	0.452 4
P7肽	-233.92	0.842 7
P7脂肽	-244.87	0.869 6