Alkhaldi Abdulsalam A M, Creek Darren J, Ibrahim Hasan, Kim Dong-Hyun, Quashie Neils B, Burgess Karl E, Changtam Chatchawan, Barrett Michael P, Suksamrarn Apichart, de Koning Harry P
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom (A.A.M.A, D.J.C., H.I., D.-H.K., N.B.Q., K.E.B., M.P.B., H.P.K.); Department of Biology, College of Science, Aljouf University, Skaka, Kingdom of Saudi Arabia (A.A.M.A); Department of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, Australia (D.J.C.); Faculty of Science, Department of Zoology, Sebha University, Libya (H.I.); Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, United Kingdom (D.-H.K.); Centre for Tropical Clinical Pharmacology and Therapeutics, University of Ghana Medical School, Accra, Ghana (N.B.Q.); Division of Physical Science, Faculty of Science and Technology, Huachiew Chalermprakiet University, Samutprakarn, Thailand (C.C.); Wellcome Trust Centre for Molecular Parasitology, University of Glasgow, Glasgow, United Kingdom (M.P.B.); and Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, Thailand (A.S.).
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom (A.A.M.A, D.J.C., H.I., D.-H.K., N.B.Q., K.E.B., M.P.B., H.P.K.); Department of Biology, College of Science, Aljouf University, Skaka, Kingdom of Saudi Arabia (A.A.M.A); Department of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, Australia (D.J.C.); Faculty of Science, Department of Zoology, Sebha University, Libya (H.I.); Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, United Kingdom (D.-H.K.); Centre for Tropical Clinical Pharmacology and Therapeutics, University of Ghana Medical School, Accra, Ghana (N.B.Q.); Division of Physical Science, Faculty of Science and Technology, Huachiew Chalermprakiet University, Samutprakarn, Thailand (C.C.); Wellcome Trust Centre for Molecular Parasitology, University of Glasgow, Glasgow, United Kingdom (M.P.B.); and Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, Thailand (A.S.)
Mol Pharmacol. 2015;87(3):451-64. doi: 10.1124/mol.114.096016. Epub 2014 Dec 19.
We have previously reported that curcumin analogs with a C7 linker bearing a C4-C5 olefinic linker with a single keto group at C3 (enone linker) display midnanomolar activity against the bloodstream form of Trypanosoma brucei. However, no clear indication of their mechanism of action or superior antiparasitic activity relative to analogs with the original di-ketone curcumin linker was apparent. To further investigate their utility as antiparasitic agents, we compare the cellular effects of curcumin and the enone linker lead compound 1,7-bis(4-hydroxy-3-methoxyphenyl)hept-4-en-3-one (AS-HK014) here. An AS-HK014-resitant line, trypanosomes adapted to AS-HK014 (TA014), was developed by in vitro exposure to the drug. Metabolomic analysis revealed that exposure to AS-HK014, but not curcumin, rapidly depleted glutathione and trypanothione in the wild-type line, although almost all other metabolites were unchanged relative to control. In TA014 cells, thiol levels were similar to untreated wild-type cells and not significantly depleted by AS-HK014. Adducts of AS-HK014 with both glutathione and trypanothione were identified in AS-HK014-exposed wild-type cells and reproduced by chemical reaction. However, adduct accumulation in sensitive cells was much lower than in resistant cells. TA014 cells did not exhibit any changes in sequence or protein levels of glutathione synthetase and γ-glutamylcysteine synthetase relative to wild-type cells. We conclude that monoenone curcuminoids have a different mode of action than curcumin, rapidly and specifically depleting thiol levels in trypanosomes by forming an adduct. This adduct may ultimately be responsible for the highly potent trypanocidal and antiparasitic activity of the monoenone curcuminoids.
我们之前报道过，带有C7连接子、C4-C5烯键连接子且在C3处有单个酮基（烯酮连接子）的姜黄素类似物，对布氏锥虫血液期形式显示出中纳摩尔活性。然而，相对于具有原始二酮姜黄素连接子的类似物，其作用机制或优越的抗寄生虫活性并无明确迹象。为了进一步研究它们作为抗寄生虫药物的效用，我们在此比较了姜黄素和烯酮连接子先导化合物1,7-双（4-羟基-3-甲氧基苯基）庚-4-烯-3-酮（AS-HK014）的细胞效应。通过体外暴露于该药物，开发出了一种对AS-HK014有抗性的细胞系，即适应了AS-HK014的锥虫（TA014）。代谢组学分析表明，暴露于AS-HK014而非姜黄素，会使野生型细胞系中的谷胱甘肽和锥虫硫醇迅速耗尽，尽管几乎所有其他代谢物相对于对照没有变化。在TA014细胞中，硫醇水平与未处理的野生型细胞相似，且不会被AS-HK014显著耗尽。在暴露于AS-HK014的野生型细胞中鉴定出了AS-HK014与谷胱甘肽和锥虫硫醇的加合物，并通过化学反应得以重现。然而，敏感细胞中的加合物积累远低于抗性细胞。相对于野生型细胞，TA014细胞在谷胱甘肽合成酶和γ-谷氨酰半胱氨酸合成酶的序列或蛋白质水平上没有表现出任何变化。我们得出结论，单烯酮姜黄素类化合物的作用模式与姜黄素不同，通过形成加合物迅速且特异性地降低锥虫中的硫醇水平。这种加合物可能最终是单烯酮姜黄素类化合物高效杀锥虫和抗寄生虫活性的原因。
