Immunoneutralization of human angiotensin-(1-12) with a monoclonal antibody in a humanized model of hypertension

Abstract

We engineered a monoclonal antibody (mAb) against the human C-terminus of angiotensin-(1–12) [h-Ang-(1–12)] and performed a biochemical characterization in concert with direct in vivo and ex vivo (carotid artery strips) assessments of h-Ang-(1–12) vasoconstrictor activity in 78 (36 females) transgenic rats expressing the human angiotensinogen gene [TGR(hAGT)L1623] and 26 (10 female) Sprague Dawley (SD) controls. The mAb shows high specificity in neutralizing angiotensin II formation from h-Ang-(1–12) and did not cross-react with human and rat angiotensins. Changes in arterial pressure and heart rate in Inactin® hydrate anesthetized rats were measured before and after h-Ang-(1–12) injections [dose range: 75–300 pmol/kg i.v.] prior to and 30–60 minutes after administration of the h-Ang-(1–12) mAb. Neutralization of circulating Ang-(1–12) inhibited the pressor action of h-Ang-(1–12), prevented Ang-(1–12) constrictor responses in carotid artery rings in both SD and TGR(hAGT)L1623 rats, and caused a fall in the arterial pressure of male and female transgenic rats. The Ang-(1–12) mAb did not affect the response of comparable dose-related pressor responses to Ang II, pre-immune IgG, or the rat sequence of Ang-(1–12). This h-Ang-(1–12) mAb can effectively suppress the pressor actions of the substrate in the circulation of hypertensive rats or in carotid artery strips from both SD and transgenic rats. The demonstration that this Ang-(1–12) mAb by itself, induced a fall in arterial pressure in transgenic hypertensive rats supports further exploring the potential abilities of Ang-(1–12) mAb in the treatment of hypertension.