Decoding the mechanism of P450-catalyzed aromatic hydroxylation:Uncovering the arene oxide pathway and insights into the regioselectivity  

解析P450催化芳环羟基化反应的机理:氧化芳烃途径以及区域选择性的探究

作　　者：Qun Huang Xuan Zhang Guangwu Sun Rui-ying Qiu Lan Luo Cuizhen Wang Longwei Gao Bing Gao Bo Chen Binju Wang Jian-bo Wang 黄群;张璇;孙光武;邱瑞英;罗兰;王翠珍;高龙威;高兵;陈波;王斌举;王健博(浙江大学医学院,“多脏器衰竭预警与干预”教育部重点实验室,浙江大学第二附属医院综合ICU及药物生物技术研究所,浙江杭州310058;湖南师范大学化学化工学院,化学生物学与中药研究教育部重点实验室,植化单体开发与利用湖南省重点实验室,湖南长沙410081;厦门大学化学化工学院,固体表面物理化学国家重点实验室,福建省理论与计算化学重点实验室,福建厦门361005;湖南大学化学化工学院,化学生物学与纳米医学研究所,化学/生物传感与化学计量学国家重点实验室,湖南长沙410082;重庆理工大学药学与生物工程学院,重庆400054;宁波大学新药技术研究院,天体化学与空间生命-钱学森空间科学协同研究中心,浙江宁波315211)

机构地区：[1]Key Laboratory of Multiple Organ Failure(Zhejiang University),Ministry of Education,Department of General Intensive Care Unit of the Second Affiliated Hospital and Institute of Pharmaceutical Biotechnology,Zhejiang University School of Medicine,Hangzhou 310058,Zhejiang,China [2]Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,Hunan,China [3]State Key Laboratory of Physical Chemistry of Solid Surfaces and Fujian Provincial Key Laboratory of Theoretical and Computational Chemistry,College of Chemistry and Chemical Engineering,Xiamen University,Xiamen361005,Fujian,China [4]State Key Laboratory of Chemo/Bio-Sensing and Chemometrics,College of Chemistry and Chemical Engineering,Institute of Chemical Biology and Nanomedicine,Hunan University,Changsha 410082,Hunan,China [5]School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,China [6]Institute of Drug Discovery Technology and Qian Xuesen Collaborative Research Center of Astrochemistry and Space Life Sciences,Ningbo University,Ningbo 315211,Zhejiang,China

出　　处：《Chinese Journal of Catalysis》2025年第3期420-430,共11页催化学报(英文)

基　　金：国家重点研发计划(2023YFA0914100/2023YFA0914102);国家自然科学基金(22077029,22477110,22034002,22276049,22073077,22001065);中央高校基本科研业务费(226202400061,226202300100);湖南省杰出青年科学基金(2021JJ10034);重庆市教委科学技术研究项目(KJQN202401148);植化单体开发与利用湖南省重点实验室开放基金(23010104).

摘　　要：P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process of aromatic hydroxylation and mechanism of regioselectivity catalyzed by cytochrome P450 monooxygenases remained ambiguous.Here,we have resolved these issues.With a stable chiral organofluorine probe,and especially with X-ray data of two isolated arene oxides derivatives,we demonstrate that an arene oxide pathway is definitely involved in P450-catalyzed aromatic hydroxylation.By the capture,isolation,identification and reactivity exploration of the arene 1,2-oxide and arene 2,3-oxide intermediates,together with advanced QM calculations,the mechanism of how two intermediates go to the same product has been elucidated.In addition to the model substrate,we also confirmed that an arene oxide intermediate is involved in the P450-catalyzed hydroxylation pathway of a natural product derivative methyl cinnamate,which indicates that this intermediate appears to be universal in P450-catalyzed aromatic hydroxylation.Our work not only provides the most direct evidence for the arene oxide pathway and new insights into the regioselectivity involved in P450-catalyzed aromatic hydroxylation,but also supplies a new synthetic approach to achieve the dearomatization of aromatic compounds.P450酶催化的芳环羟基化在芳香族化合物的解毒、生物合成和潜在致癌作用中均起着重要作用.虽然其催化机理与烷烃的羟基化一样,是由高反应活性的血红素-Fe(IV)=O(Cpd I)介导,但相比之下,Cpd I与芳香族π系统的反应更为复杂.尽管P450催化芳环羟基化的机理被研究了几十年,目前仍存在许多争议.此外,实验表明取代芳香族化合物的羟基化通常发生在邻位或对位,该区域选择性是否与已知或未知中间体的生成有关,还没有相关研究.因此,有关P450催化芳环羟基化的真实过程以及机理细节仍有待深入研究.有机分子中,氟元素的引入可以显著提高化合物的代谢稳定性,据此我们推测含氟芳香族化合物,可能是P450催化芳环羟基化反应中获得稳定中间体的合适底物.因此,本文以α-氟代苯乙酸乙酯为化学探针,P450-BM3为催化剂,以期检测出相应的关键中间体,详细阐明P450催化芳环羟基化的机理.通过对α-氟代苯乙酸乙酯转化体系进行反应时间尺度监测,

关 键 词：Aromatic hydroxylation Cytochrome P450 Arene oxide Regioselectivity DEAROMATIZATION 

分 类 号：O621.2[理学—有机化学] O62[理学—化学]