PC18, a specific aminopeptidase N inhibitor, induces vasopressin release by increasing the half-life of brain angiotensin III.
Angiotensin III (AngIII), which is metabolized in vivo by aminopeptidase N (APN), was previously shown to be one of the main effector peptides of the brain renin-angiotensin system (RAS) in the control of vasopressin release. Recently, a potent APN inhibitor, PC18 (2-amino-4-methylsulfonyl butane thiol, methionine thiol), has been developed. In this study, we first checked the in vitro selectivity of PC18 towards APN, aminopeptidase A (APA) and aminopeptidase B (APB), three zinc metalloproteases with significant identity between their amino acid sequences. The Ki values of this compound on APN were found to be in the nanomolar range (Ki = 8.0 +/- 1.7 nM) but it was 2,150 and 125 times less active on APA and APB, respectively. Secondly, we evaluated in vivo the effect of brain APN inhibition with PC18 on the inactivation of brain AngIII and on vasopressin secretion in mice. For this purpose, mice received [3H]AngII intracerebroventricularly in the presence or absence of the APN inhibitor PC18 (30 microg). At different times after the injection, [3H]AngIII levels were evaluated from hypothalamus homogenates after separation by cation-exchange chromatography. PC18 induced an accumulation of [3H]AngIII, increasing its half-life 3.9 times as compared with control values. In addition, the effect of PC18 on vasopressin release was studied in mice. PC18 (10-100 microgram) was injected intracerebroventricularly, and plasma vasopressin levels were estimated by radioimmunoassay. PC18 increased vasopressin levels in a dose-dependent manner. The maximal increase in vasopressin release (+220%) is observed for a dose of PC18 of 100 microgram and was inhibited 75% by the coadministration of the AngII receptor antagonist (Sar1-Ala8)-AngII (0.5 microgram). These results indicate that in vivo, in the mouse brain, APN inhibition by PC18 increases the half-life of endogenous AngIII, resulting in an enhanced vasopressin release.
Réaux
A
A
Institut National de la Santé et de la Recherche Médicale, Unité 36, Collège de France, Paris, France.
de Mota
N
N
Zini
S
S
Cadel
S
S
Fournié-Zaluski
M C
MC
Roques
B P
BP
Corvol
P
P
Llorens-Cortès
C
C
D000626:Aminopeptidases / Q000037:antagonists & inhibitors; D000804:Angiotensin II / Q000096:biosynthesis / Q000302:isolation & purification; D000805:Angiotensin III / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001923:Brain Chemistry / Q000187:drug effects*; D018826:CD13 Antigens / Q000037:antagonists & inhibitors*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D043384:Glutamyl Aminopeptidase; D006207:Half-Life; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism; D007276:Injections, Intraventricular; D008297:Male; D051379:Mice; D011480:Protease Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D014667:Vasopressins / Q000097:blood / Q000378:metabolism*
D011480:Protease Inhibitors; D014667:Vasopressins; D000804:Angiotensin II; D000805:Angiotensin III; D000626:Aminopeptidases; D018826:CD13 Antigens; C019721:aminopeptidase B; D043384:Glutamyl Aminopeptidase