Kintos Dionysios-Panagiotis, Salagiannis Konstantinos, Vazoura Vasiliki, Wittrien Theresa, Papakyriakou Athanasios, Nikolaropoulos Sotiris S, Behrends Soenke, Topouzis Stavros, Fousteris Manolis A
Laboratory of Medicinal Chemistry, Department of Pharmacy, University of Patras, Patras, GR-26500, Greece.
Laboratory of Molecular Pharmacology, Department of Pharmacy, University of Patras, Patras, GR-26500, Greece.
Heliyon. 2022 Nov 6;8(11):e11438. doi: 10.1016/j.heliyon.2022.e11438. eCollection 2022 Nov.
Herein, we present the structure-based design, synthesis and biological evaluation of novel mono- and di-carboxylic 3,4-dihydroquinoxalin-2(1)-one derivatives as potential heme-independent activators of soluble guanylate cyclase (sGC). Docking calculations of several known sGC agonists by utilizing both a homology model of human sGC Η-ΝΟΧ domain and a recent cryo-EM structure of the same domain guided the structural optimization of various designed compounds. Among these, mono- and di-carboxylic 3,4-dihydroquinoxalin-2(1)-one derivatives arose as promising candidate sGC activators. A series of such compounds was synthesized and assessed for their effect on sGC activity. None of them was able to trigger any detectable activation of native sGC in prostate cancer (LnCaP) or rat aortic smooth muscle (A7r5) cells, even after loss of heme by treatment with the heme oxidant ODQ. Furthermore, selected derivatives did not exhibit any antagonistic effect against the known heme-independent sGC activator BAY 60-2770 nor any additive or synergistic effect with the heme-dependent NO donor sodium nitroprusside (SNP) on heme-associated sGC in A7r5 cells. However, when tested using purified recombinant sGC enzyme, the dicarboxylic 3,4-dihydroquinoxalin-2(1)-one derivative was able to increase the enzymatic activity of both the wild-type α1/β1 sGC dimer (by 4.4-fold, EC = 0.77 μΜ) as well as the heme-free α1/β1 His105Ala mutant sGC (by 4.8-fold, EC = 1.8 μΜ). Notably, the activity of compound towards the mutant α1/β1 Η105A enzyme was comparable with that previously reported by us for the activator BAY 60-2770, using the functionally equivalent wild-type sGC preparation treated with ODQ. These results indicate that compound can indeed act as a promising sGC activator and may serve as a basic structure in the design of novel, optimized analogues with enhanced sGC agonistic activity and improved efficiency in cell-based and systems.
在此，我们展示了新型单羧酸和二羧酸3,4 - 二氢喹喔啉 - 2(1) - 酮衍生物作为可溶性鸟苷酸环化酶（sGC）潜在的非血红素依赖性激活剂的基于结构的设计、合成及生物学评价。利用人类sGC Η - ΝΟΧ结构域的同源模型和该结构域最近的冷冻电镜结构对几种已知的sGC激动剂进行对接计算，指导了各种设计化合物的结构优化。其中，单羧酸和二羧酸3,4 - 二氢喹喔啉 - 2(1) - 酮衍生物成为有前景的sGC激活剂候选物。合成了一系列此类化合物，并评估了它们对sGC活性的影响。即使在用血红素氧化剂ODQ处理使血红素丧失后，它们中没有一个能够在前列腺癌（LnCaP）或大鼠主动脉平滑肌（A7r5）细胞中引发天然sGC的任何可检测到的激活。此外，所选衍生物对已知的非血红素依赖性sGC激活剂BAY 60 - 2770没有表现出任何拮抗作用，在A7r5细胞中对与血红素相关的sGC也没有与血红素依赖性一氧化氮供体硝普钠（SNP）产生任何相加或协同作用。然而，当使用纯化的重组sGC酶进行测试时，二羧酸3,4 - 二氢喹喔啉 - 2(1) - 酮衍生物能够增加野生型α1/β1 sGC二聚体的酶活性（增加4.4倍，EC = 0.77 μΜ）以及无血红素的α1/β1 His105Ala突变型sGC的酶活性（增加4.8倍，EC = 1.8 μΜ）。值得注意的是，该化合物对突变型α1/β1 Η105A酶的活性与我们之前使用用ODQ处理的功能等效的野生型sGC制剂报道的激活剂BAY 60 - 2770相当。这些结果表明该化合物确实可以作为一种有前景的sGC激活剂，并可能作为设计具有增强的sGC激动活性和在基于细胞及系统中提高效率的新型优化类似物的基本结构。
