Yeh Tzu-Lan, Leissing Thomas M, Abboud Martine I, Thinnes Cyrille C, Atasoylu Onur, Holt-Martyn James P, Zhang Dong, Tumber Anthony, Lippl Kerstin, Lohans Christopher T, Leung Ivanhoe K H, Morcrette Helen, Clifton Ian J, Claridge Timothy D W, Kawamura Akane, Flashman Emily, Lu Xin, Ratcliffe Peter J, Chowdhury Rasheduzzaman, Pugh Christopher W, Schofield Christopher J
Chemistry Research Laboratory , Department of Chemistry , University of Oxford , Oxford OX1 3TA , UK . Email:
Target Discovery Institute (TDI) , Nuffield Department of Medicine , University of Oxford , NDMRB Roosevelt Drive , Oxford OX3 7FZ , UK.
Chem Sci. 2017 Nov 1;8(11):7651-7668. doi: 10.1039/c7sc02103h. Epub 2017 Sep 11.
Inhibition of the human 2-oxoglutarate (2OG) dependent hypoxia inducible factor (HIF) prolyl hydroxylases (human PHD1-3) causes upregulation of HIF, thus promoting erythropoiesis and is therefore of therapeutic interest. We describe cellular, biophysical, and biochemical studies comparing four PHD inhibitors currently in clinical trials for anaemia treatment, that describe their mechanisms of action, potency against isolated enzymes and in cells, and selectivities representatives of other human 2OG oxygenase subfamilies. The 'clinical' PHD inhibitors are potent inhibitors of PHD catalyzed hydroxylation of the HIF-α oxygen dependent degradation domains (ODDs), and selective against most, but not all, representatives of other human 2OG dependent dioxygenase subfamilies. Crystallographic and NMR studies provide insights into the different active site binding modes of the inhibitors. Cell-based results reveal the inhibitors have similar effects on the upregulation of HIF target genes, but differ in the kinetics of their effects and in extent of inhibition of hydroxylation of the N- and C-terminal ODDs; the latter differences correlate with the biophysical observations.
抑制人源2-氧代戊二酸（2OG）依赖性缺氧诱导因子（HIF）脯氨酰羟化酶（人源PHD1-3）会导致HIF上调，从而促进红细胞生成，因此具有治疗意义。我们描述了细胞、生物物理和生化研究，比较了目前正在进行贫血治疗临床试验的四种PHD抑制剂，阐述了它们的作用机制、对分离酶和细胞的效力，以及对其他人类2OG加氧酶亚家族代表的选择性。“临床用”PHD抑制剂是PHD催化的HIF-α氧依赖性降解结构域（ODD）羟基化的有效抑制剂，对其他人类2OG依赖性双加氧酶亚家族的大多数（但不是全部）代表具有选择性。晶体学和核磁共振研究揭示了抑制剂不同的活性位点结合模式。基于细胞的结果表明，这些抑制剂对HIF靶基因的上调具有相似的作用，但在作用动力学以及对N端和C端ODD羟基化的抑制程度上存在差异；后一种差异与生物物理观察结果相关。
