Wiemer Gabriele, Dobrucki Lawrence W, Louka Febee R, Malinski Tadeusz, Heitsch Holger
DG Cardiovascular Diseases/Medicinal Chemistry, Aventis Pharma Deutschland GmbH, Frankfurt/Main, Germany.
Hypertension. 2002 Dec;40(6):847-52. doi: 10.1161/01.hyp.0000037979.53963.8f.
Recently, we demonstrated that the heptapeptide angiotensin-(1-7) (Ang-[1-7]) exhibits a favorable kinetic of nitric oxide (NO) release accompanied by extremely low superoxide (O2-) production. In this report we describe AVE 0991, a novel nonpeptide compound that evoked effects similar to Ang-(1-7) on the endothelium. AVE 0991 and unlabeled Ang-(1-7) competed for high-affinity binding of [125I]-Ang-(1-7) to bovine aortic endothelial cell membranes with IC50 values of 21+/-35 and 220+/-280 nmol/L, respectively. Stimulated NO and O2- release from bovine aortic endothelial cells was directly and simultaneously measured on the cell surface by selective electrochemical nanosensors. Peak concentrations of NO and O2- release by AVE 0991 and Ang-(1-7) (both 10 micromol/L) were not significantly different (NO: 295+/-20 and 270+/-25 nmol/L; O2-: 18+/-2 and 20+/-4 nmol/L). However, the released amount of bioactive NO was approximately 5 times higher for AVE 0991 in comparison to Ang-(1-7). The selective Ang-(1-7) antagonist [D-Ala(7)]-Ang-(1-7) inhibited the AVE 0991-induced NO and O2- production by approximately 50%. A similar inhibition level was observed for the Ang II AT1 receptor antagonist EXP 3174. In contrast, the Ang II AT2 receptor antagonist PD 123,177 inhibited the AVE 0991-stimulated NO production by approximately 90% but without any inhibitory effect on O2- production. Both NO and O2- production were inhibited by NO synthase inhibition ( approximately 70%) and by bradykinin B2 receptor blockade (approximately 80%). AVE 0991 efficiently mimics the effects of Ang-(1-7) on the endothelium, most probably through stimulation of a specific, endothelial Ang-(1-7)-sensitive binding site causing kinin-mediated activation of endothelial NO synthase.
最近，我们证明七肽血管紧张素 -（1 - 7）（Ang - [1 - 7]）表现出有利的一氧化氮（NO）释放动力学，同时超氧化物（O₂⁻）产生极低。在本报告中，我们描述了AVE 0991，一种新型非肽化合物，它在内皮细胞上引发了与Ang -（1 - 7）相似的效应。AVE 0991和未标记的Ang -（1 - 7）竞争[¹²⁵I]-Ang -（1 - 7）与牛主动脉内皮细胞膜的高亲和力结合，IC₅₀值分别为21±35和220±280 nmol/L。通过选择性电化学纳米传感器在细胞表面直接同时测量牛主动脉内皮细胞刺激后的NO和O₂⁻释放。AVE 0991和Ang -（1 - 7）（均为10 μmol/L）释放的NO和O₂⁻的峰值浓度无显著差异（NO：295±20和270±25 nmol/L；O₂⁻：18±2和20±4 nmol/L）。然而，与Ang -（1 - 7）相比，AVE 0991释放的生物活性NO量大约高5倍。选择性Ang -（1 - 7）拮抗剂[D - Ala(7)]-Ang -（1 - 7）将AVE 0991诱导的NO和O₂⁻产生抑制了约50%。血管紧张素II AT1受体拮抗剂EXP 3174也观察到类似的抑制水平。相比之下，血管紧张素II AT2受体拮抗剂PD 123,177将AVE 0991刺激的NO产生抑制了约90%，但对O₂⁻产生没有任何抑制作用。NO合酶抑制（约70%）和缓激肽B2受体阻断（约80%）均抑制了NO和O₂⁻的产生。AVE 0991有效地模拟了Ang -（1 - 7）对内皮细胞的作用，很可能是通过刺激一个特定的、对内皮细胞Ang -（1 - 7）敏感的结合位点，导致激肽介导的内皮型一氧化氮合酶激活。
