Experimental and Clinical Pharmacology Research Group, Department of Medical Biology, UiT, The Arctic University of Norway, 9037 Tromsø, Norway.
Experimental and Clinical Pharmacology Research Group, Department of Medical Biology, UiT, The Arctic University of Norway, 9037 Tromsø, Norway; Department of Clinical Pharmacology, Division of Diagnostic Services, University Hospital of North Norway, 9038 Tromsø, Norway.
Biomed Pharmacother. 2020 Jun;126:110109. doi: 10.1016/j.biopha.2020.110109. Epub 2020 Mar 28.
Clinical studies have reported overexpression of PDE5 and elevation of intracellular cyclic GMP in various types of cancer cells. ABCC5 transports cGMP out of the cells with high affinity. PDE5 inhibitors prevent both cellular metabolism and cGMP efflux by inhibiting ABCC5 as well as PDE5. Increasing intracellular cGMP is hypothesized to promote apoptosis and growth restriction in tumor cells and also has potential for clinical use in treatment of cardiovascular disease and erectile dysfunction. Vardenafil is a potent inhibitor of both PDE5 and ABCC5-mediated cGMP cellular efflux. Nineteen novel vardenafil analogs that have been predicted as potent inhibitors by VLS were chosen for tests of their ability to inhibit ATP- dependent transport of cGMP by measuring the accumulation of cyclic GMP in inside-out vesicles.
In this study, we investigated the ability of nineteen new compounds to inhibit ABCC5- mediated cGMP transport. We also determined the Ki values of the six most potent compounds.
Preparation of human erythrocyte inside out vesicles and transport assay.
Ki values for six of nineteen compounds that showed more than 50 % inhibition of cGMP transport in the screening test were determined and ranged from 1.1 to 23.1 μM. One compound was significantly more potent than the positive control, sildenafil.
Our findings show that computational screening correctly identified vardenafil-analogues that potently inhibit cGMP efflux-pumps from cytosol and could have substantial clinical potential in treatment of patients with diverse disorders.
临床研究报告称，在各种类型的癌细胞中 PDE5 的表达过度和细胞内环鸟苷酸 (cGMP) 的升高。ABCC5 以高亲和力将 cGMP 从细胞内转运出去。PDE5 抑制剂通过抑制 ABCC5 和 PDE5 来防止细胞内代谢和 cGMP 外排。增加细胞内 cGMP 被假设为促进肿瘤细胞凋亡和生长受限，并且也有可能在治疗心血管疾病和勃起功能障碍方面的临床应用。伐地那非是一种强效的 PDE5 和 ABCC5 介导的 cGMP 细胞外排抑制剂。通过 VLS 预测为强效抑制剂的 19 种新型伐地那非类似物被选择用于测试其抑制由 ATP 依赖性 cGMP 转运的能力，方法是测量无细胞囊泡中环鸟苷酸的积累。
在这项研究中，我们研究了 19 种新化合物抑制 ABCC5 介导的 cGMP 转运的能力。我们还确定了 6 种最有效化合物的 Ki 值。
人红细胞外翻小体的制备和转运测定。
在筛选试验中显示 cGMP 转运抑制超过 50%的 19 种化合物中的 6 种的 Ki 值被确定，范围从 1.1 到 23.1 μM。一种化合物比阳性对照西地那非明显更有效。
我们的发现表明，计算筛选正确地鉴定了伐地那非类似物，它们能有效地抑制细胞溶质中的 cGMP 外排泵，并且在治疗患有各种疾病的患者方面可能具有重要的临床潜力。
