2-Amino-nonyl-6-methoxyl-tetralin muriate inhibits sterol C-14 reductase in the ergosterol biosynthetic pathway  

作　　者：Rong-mei LIANG Yong-bing CAO Kai-hua FAN Yi XU Ping-hui GAO You-jun ZHOU Bao-di DAI Yong-hong TAN Shi-hua WANG Hui TANG Hong-tao LIU Yuan-ying JIANG 

机构地区：[1]Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China [2]Department of Pharmacy, General Hospital of Chengdu Military Command Region, Chengdu 610083, China [3]Department of Medicinal Chemistry, School of Pharmacy, Second Military Medical University, Shanghai 200433, China [4]Department of Pharmacology, School of Life Sci- ence and Biopharmacology, Shenyang Pharmaceutical University, Shenyang 110016, China [5]Department of Pharmacy, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, China

出　　处：《Acta Pharmacologica Sinica》2009年第12期1709-1716,共8页中国药理学报（英文版）

基　　金：Acknowledgments Project was supported by the National Natural Science Foundation of China （No 30672626 and 30572257）. We thank Dr Joseph HEITMAN for kindly providing the isolate C albicans strain BWP17 and NJ52-1 and Dr William A FONZI for kindly offering the isolate C albicans SC5314 in this study.

摘　　要：Aim： To investigate the action mechanism of a novel chemical structural aminotetralin derivate, 2-Amino-Nonyl-6-MethoxyI-Tetralin Muriate （lOb）, against Candida albicans （C albicans） in the ergosterol biosynthetic pathway. Methods： Antifungal susceptibility test of lOb was carried out using broth microdilution method, the action mechanism of lOb against C albicans was investigated by GC-MS spectrometry and real-time RT-PCR assay, and cytotoxicity of lOb in vitro was assessed by MTS/ PMS reduction assay. Results： 10b reduced the ergosterol content markedly, and the 50% ergosterol content inhibitory concentration （ECIC50 value） was 0.08 μg/mL. Although the sterol composition of 10b-grown cells was completely identical with that of erg24 strain, the content of ergosta- 8,14,22-trienol in 10b-grown cells was much higher than that in erg24 strain. Real-time RT-PCR assay revealed a global upregulation of sterol metabolism genes. In addition, the 50% inhibitory concentration （IC50 value） of lOb was 11.30 μg/mL for murine embryonic fibroblasts and 35.70 μg/mL for human normal liver cells. Conclusion： lOb possessed a mode of action different from that of azoles and morpholines, whose targets were sterol C-14 reductase （encoded by ERG24 gene） and sterol C-5 desaturase （encoded by ERG3） related enzyme. Although lOb seemed to reduce MTS/PMS reduction in a dose dependent manner, IC50 value for mammalian cells was much higher than 50% minimum inhibitory concentration （MIC50） value for C albicans. This indicates that the formulation is preliminarily safe and warrants further study for possible human applications.Aim： To investigate the action mechanism of a novel chemical structural aminotetralin derivate, 2-Amino-Nonyl-6-MethoxyI-Tetralin Muriate （lOb）, against Candida albicans （C albicans） in the ergosterol biosynthetic pathway. Methods： Antifungal susceptibility test of lOb was carried out using broth microdilution method, the action mechanism of lOb against C albicans was investigated by GC-MS spectrometry and real-time RT-PCR assay, and cytotoxicity of lOb in vitro was assessed by MTS/ PMS reduction assay. Results： 10b reduced the ergosterol content markedly, and the 50% ergosterol content inhibitory concentration （ECIC50 value） was 0.08 μg/mL. Although the sterol composition of 10b-grown cells was completely identical with that of erg24 strain, the content of ergosta- 8,14,22-trienol in 10b-grown cells was much higher than that in erg24 strain. Real-time RT-PCR assay revealed a global upregulation of sterol metabolism genes. In addition, the 50% inhibitory concentration （IC50 value） of lOb was 11.30 μg/mL for murine embryonic fibroblasts and 35.70 μg/mL for human normal liver cells. Conclusion： lOb possessed a mode of action different from that of azoles and morpholines, whose targets were sterol C-14 reductase （encoded by ERG24 gene） and sterol C-5 desaturase （encoded by ERG3） related enzyme. Although lOb seemed to reduce MTS/PMS reduction in a dose dependent manner, IC50 value for mammalian cells was much higher than 50% minimum inhibitory concentration （MIC50） value for C albicans. This indicates that the formulation is preliminarily safe and warrants further study for possible human applications.

关 键 词：Candida albicans sterol C-14 reducase ERGOSTEROL 2-amino-nonyl-6-methoxyl-tetralin muriate 

分 类 号：Q946.88[生物学—植物学] TQ923[轻工技术与工程—发酵工程]