Q79
山东省重点研发计划（重大科技创新工程）（编号：2021CXGC010506）
目的 对μ-CnIIIC芋螺多肽的3种二硫键异构体进行合成鉴定及生物安全性、活性评价。方法 通过固相多肽合成法（SPPS）人工合成μ-CnIIIC-Ⅰ、μ-nIIIC-Ⅱ和μ-CnIIIC-Ⅲ线性肽；采用氧化法对线性肽进行氧化折叠，获得3种折叠肽异构体（μ-CnIIIC-Ⅰ、μ-CnIIIC-Ⅱ和μ-CnIIIC-Ⅲ），并对其进行质谱表征和HPLC纯化；通过MTT法测试3种异构体的细胞毒性；采用双电极电压膜片钳技术测试3种异构体对NaV1.7的抑制作用。结果 质谱表征结果显示成功合成μ-nII?????????????????ǘ???????ǘ???????????????????????ǘ???????????????ǘ???????ǘ???????????????????????ǘ???????????????ǘ???????ǘ???????????????????????ǘ???????????????ǘ???????ǘ???????????????????????ǘ?????????
Objective To study the synthesis, identification and activity of three disulfide bond isomers of μ-CnIIIC. Methods The linear peptides of μ-CnIIIC-Ⅰ, μ-CnIIIC-Ⅱ and μ-CnIIIC-Ⅲ were synthesized by solid phase peptide synthesis (SPPS). Three-fold peptide isomers (μ-CnIIIC-Ⅰ, μ-CnIIIC-Ⅱ and μ-CnIIIC-Ⅲ) were obtained by oxidative folding of the linear peptides, which were characterized by mass spectrometry and purified by HPLC. The cytotoxicity of the three isomers was tested by MTT method. The inhibition effect of three isomers on NaV1.2 was tested by using double electrode voltage patch clamp technique. Results Mass spectrometry characterization results showed that μ-CnIIIC-Ⅰ, μ-CnIIIC-Ⅱ and μ-CnIIIC-Ⅲ were successfully synthesized. When the concentration of three isomers of μ-CnIIIC was in the range of 50 μmol/L, the cell survival rate was above 75% and no hemolysis occurred. When the concentration of μ-CnIIIC isomer was 20 μmol/L, the inhibition rates of μ-CnIIIC-Ⅰ, μ-CnIIIC-Ⅱ and μ-CnIIIC-Ⅲ on sodium channel were 23.64%, 52.43% and 81.21%, respectively. The average scores of skin irritation test were 0.107, 0.179 and 0.286 per animal per day. Conclusion The μ-CnIIIC isomers of conopeptides have good biosafety. The μ-CnIIIC isomers with disulfide bonds of Ⅰ-Ⅳ, Ⅱ-Ⅴ and Ⅲ-Ⅵ have better selectivity to sodium channels, stronger inhibitory effect and better biological activity.
姬胜利,李丹,吴伟东,等.μ-CnIIIC 芋螺多肽二硫键异构体的合成鉴定及活性评价[J].医学美学美容,2023,32(2):6-10.
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