基于自组装体系研究双黄连主要成分与环丙沙星的分子互作

Molecular interactions between the main components of Shuanghuanglian injection and ciprofloxacin injection based on self-assembly

双黄连注射液(双黄连)与环丙沙星注射液(环丙沙星)在临床联合用药的情况时有发生,但联合用药合理性评价手段薄弱。本研究采用等温滴定量热技术和紫外光谱初步发现双黄连及其主要成分绿原酸、新绿原酸与环丙沙星均存在分子互作;采用扫描电镜、红外光谱、冷喷雾电离质谱技术证实这种分子互作与绿原酸、新绿原酸能和环丙沙星通过分子间弱键诱导形成自组装超分子体系有关;进一步利用铜绿假单胞菌评价其药物分子互作生物效应(抑菌活性),发现双黄连、绿原酸、新绿原酸与环丙沙星互作后对铜绿假单胞菌的抑菌能力均显著降低;通过分子对接研究发现,其抑菌能力降低与药物分子在铜绿假单胞菌DNA旋转酶B亚单位(DNA gyrase B,GyrB)有相同结合位点而产生竞争性结合密切相关。本研究从分子自组装角度,揭示了双黄连及其主要成分绿原酸、新绿原酸与环丙沙星存在分子间互作且会导致抑菌能力降低,为双黄连与环丙沙星临床联合用药合理性评价提供了参考依据。

The combination of Shuanghuanglian injection(SHLI)and ciprofloxacin injection(CIPI)is frequently prescribed in clinical practice,but the basis for the combination is weak.In this study,isothermal titration calorimetry and ultraviolet-visible absorption spectrometry were applied to identify the molecular interactions of SHLI and its main components,chlorogenic acid and neochlorogenic acid with CIPI.Scanning electron microscopy,Fouriertransform infrared spectroscopy,and cold-spray ionization mass spectrometry were performed to confirm that this molecular interaction was related to the formation of self-assembled supramolecular systems induced by chlorogenic acid and neochlorogenic acid with CIPI through weak intermolecular bonds.The antibacterial activity toward Pseudomonas aeruginosa(P.aeruginosa)was evaluated via molecular interactions,and the inhibitory ability of SHLI,chlorogenic acid and neochlorogenic acid against P.aeruginosa was significantly reduced after interaction with CIPI.A molecular docking study demonstrated that the reduced antibacterial ability was closely related to the competitive binding of drug molecules to the same binding site of the DNA gyrase B(GyrB)subunit of P.aeruginosa.The present study uncovered the intermolecular interactions of SHLI and its main components chlorogenic acid and neochlorogenic acid with CIPI from the perspective of molecular self-assembly and contribute to the reduction of its antibacterial ability,providing a basis for the clinical combination of SHLI and CIPI.