Michael acceptor containing drugs are a novel class of 5-lipoxygenase inhibitor targeting the surface cysteines C416 and C418

详细信息    查看全文

• 作者：Isabelle V. Maucher^a ; Michael Rü ; hl^a ; Simon B.M. Kretschmer^a ; Bettina Hofmann^a ; Benjamin Kü ; hn^a ; Jasmin Fettel^a ; Anja Vogel^a ; ^b ; Karsten T. Flü ; gel^a ; Georg Manolikakes^c ; Nadine Hellmuth^a ; Ann-Kathrin Hä ; fner^a ; Vahid Golghalyani^a ; Ann-Katrin Ball^a ; Matthias Piesche^d ; ^g ; Carmela Matrone^d ; Gerd Geisslinger^b ; ^e ; Michael J. Parnham^b ; Michael Karas^a ; Dieter Steinhilber^a ; Jessica Roos^a ; ¹ ; ^{roos@pharmchem.uni-frankfurt.de} ; Thorsten J. Maier^d ; ^f ; ¹ ; ^{maier@biomed.au.dk}
• 关键词：AA ; arachidonic acid ; methyl-BQ ; 2 ; 5-dimethyl-1 ; 4-benzoquinone ; CDDO ; CDDO-methylester ; methoxy-BQ ; 2 ; 3-dimethoxy-5-methyl-p-benzoquinone ; methoxy-NS ; 3-methoxy-β-nitrostyrene ; DMF ; dimethyl fumarate ; GSH ; glutathione ; OH-BQ ; 2 ; 5-dihydroxy-1 ; 4-benzoquinone ; HPLC ; high-performance liquid chromatography ; H(P)ETE ; hydro (peroxy-) eicosatetraenoic acid ; LT ; leukotrienes ; MS ; mass spectrometry ; NAC ; N-acetylcysteine ; NAPQI ; N-acetyl-p-benzoquinone imine ; NEM ; N-ethylmaleimide ; NFA ; nitro-fatty acid
• 刊名：Biochemical Pharmacology
• 出版年：2017
• 出版时间：1 February 2017
• 年：2017
• 卷：125
• 期：Complete
• 页码：55-74
• 全文大小：3425 K
• 卷排序：125

文摘

Recently, we published that nitro-fatty acids (NFA) are potent electrophilic molecules which inhibit 5-lipoxygenase (5-LO) by interacting catalytically with cysteine residues next to a substrate entry channel. The electrophilicity is derived from an intramolecular Michael acceptor moiety consisting of an electron-withdrawing group in close proximity to a double bond. The potential of the Michael acceptor moiety to interact with functionally relevant cysteines of proteins potentially renders them effective and sustained enzyme activity modulators. We screened a large library of naturally derived and synthetic electrophilic compounds to investigate whether other types of Michael acceptor containing drugs suppress 5-LO enzyme activity. The activity was measured by assessing the effect on the 5-LO product formation of intact human polymorphonuclear leukocytes. We demonstrated that a number of structurally different compounds were suppressive in the activity assays and showed that Michael acceptors of the quinone and nitro-alkene group produced the strongest inhibition of 5-LO product formation. Reactivity with the catalytically relevant cysteines 416 and 418 was confirmed using mutated recombinant 5-LO and mass spectrometric analysis (MALDI-MS). In the present study, we show for the first time that a number of well-recognized naturally occurring or synthetic anti-inflammatory compounds carrying a Michael acceptor, such as thymoquinone (TQ), the paracetamol metabolite NAPQI, the 5-LO inhibitor AA-861, and bardoxolone methyl (also known as RTA 402 or CDDO-methyl ester) are direct covalent 5-LO enzyme inhibitors that target the catalytically relevant cysteines 416 and 418.