State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu, PR China.
PLoS One. 2014 Apr 22;9(4):e95692. doi: 10.1371/journal.pone.0095692. eCollection 2014.
Hypoxia inducible factor 1 (HIF-1) plays a pivotal role in cellular responses to hypoxia. Prolyl hydroxylase 3 (PHD3) degrades HIF-1α under normoxic conditions through the hydroxylation of HIF-1α for proteolysis. Inhibiting PHD3 activity is crucial for up-regulating HIF-1α, thereby acting as a potential target for treating hypoxia-related diseases. In this study, two proline analogues (PA1 and PA2) were screened as PHD3 inhibitors with apparent EC50 values of 1.53 and 3.17 µM respectively, indicating good inhibition potency. Nine proteins, significantly regulated by PA1, were identified using 2-DE coupled with MALDI-TOF/TOF MS. Pyruvate kinase isozymes M1/M2 (PKM) and alpha-enolase 1 (ENO1), which are key modulators of glycolysis, are directly regulated by HIF-1α. Moreover, VEGF, a signal protein stimulating angiogenesis, was strongly promoted by PA1. Our findings suggest that PA1 stabilized HIF-1α as well as up-regulated glycolysis and angiogenesis proteins. Herein, for the first time, we systematically studied proline analogue PA1 as a PHD3 inhibitor, which provides innovative evidence for the treatment of HIF-related diseases.
缺氧诱导因子 1（HIF-1）在细胞对缺氧的反应中起着关键作用。脯氨酰羟化酶 3（PHD3）通过对 HIF-1α 的羟化作用使其发生蛋白水解，在常氧条件下降解 HIF-1α。抑制 PHD3 的活性对于上调 HIF-1α至关重要，因此它是治疗缺氧相关疾病的潜在靶标。在这项研究中，两种脯氨酸类似物（PA1 和 PA2）被筛选为 PHD3 抑制剂，其 EC50 值分别为 1.53 和 3.17 μM，表明具有良好的抑制能力。使用 2-DE 结合 MALDI-TOF/TOF MS 鉴定了 9 种明显受 PA1 调控的蛋白质。丙酮酸激酶同工酶 M1/M2（PKM）和α-烯醇酶 1（ENO1）是糖酵解的关键调节剂，它们直接受到 HIF-1α的调节。此外，PA1 强烈促进了血管生成的信号蛋白 VEGF。我们的研究结果表明，PA1 稳定了 HIF-1α，同时上调了糖酵解和血管生成蛋白。在此，我们首次系统地研究了脯氨酸类似物 PA1 作为 PHD3 抑制剂，为 HIF 相关疾病的治疗提供了创新性的证据。
