Decarboxylation of Ang-(1–7) to Ala¹-Ang-(1–7) leads to significant changes in pharmacodynamics

The heptapeptide angiotensin (Ang)-(1–7) is part of the beneficial arm of the renin-angiotensin system. Ang-(1–7) has cardiovascular protective effects, stimulates regeneration, and opposes the often detrimental effects of AngII. We recently identified the G protein-coupled receptors Mas and MrgD as receptors for the heptapeptide. Ala¹-Ang-(1–7) (Alamandine), a decarboxylated form of Ang-(1–7), has similar vasorelaxant effects, but has been described as only stimulating MrgD. Therefore, this study aimed to characterise the consequences of the lack of the carboxyl group in amino acid 1 on intracellular signalling and to identify the receptor fingerprint for Ala¹-Ang-(1–7). In primary endothelial and mesangial cells, Ala¹-Ang-(1–7) elevated cAMP concentration. Dose response curves generated with Ang-(1–7) and Ala¹-Ang-(1–7) significantly differed from each other, with a much lower EC₅₀ and a bell-shape curve for Ala¹-Ang-(1–7). We provided pharmacological proof that both, Mas and MrgD, are functional receptors for Ala¹-Ang-(1–7). Consequently, in primary mesangial cells with genetic deficiency in both receptors, the heptapeptide failed to increase cAMP concentration. As we previously described for Ang-(1–7), the Ala¹-Ang-(1–7)-mediated cAMP increase in Mas/MrgD-transfected HEK293 cells and primary cells was blocked by the AT2 receptor blocker, PD123319. The very distinct dose-response curves for both heptapeptides could be explained by in silico modelling, electrostatic potential calculations, and an involvement of G_{alpha i} for higher concentrations of Ala¹-Ang-(1–7). Our results identify Ala¹-Ang-(1–7) as a peptide with specific pharmacodynamic properties and builds the basis for the design of more potent and efficient Ang-(1–7) analogues for therapeutic intervention in a rapidly growing number of diseases.