Functional studies but not receptor binding can distinguish surmountable from insurmountable AT1 antagonism.
Our study demonstrated that inhibition of angiotensin II- (Ang II) mediated contractions of rabbit aorta by structurally diverse nonpeptide AT1 antagonists could distinguish surmountable from insurmountable AT1 antagonism. CI-996, L158809, EXP 3174 and SKF 108834 produced concentration-related rightward shifts in Ang II response curves and reduced the maximal contraction to Ang II, characteristic of insurmountable antagonism. In contrast, DuP 753 and SKF 108566, produced parallel rightward shifts in Ang II contractile curves without affecting the maximal response which is consistent with the definition of surmountable or competitive antagonism. In addition, CI-996 demonstrated potent inhibition of Ang II-stimulated inositol phosphate accumulation in rat aortic smooth muscle cells, behaving as an insurmountable antagonist. However, DuP 753 was a surmountable antagonist of Ang II-stimulated inositol phosphate accumulation. Repeated washing of rabbit aorta preincubated with either CI-996 or EXP 3174 did not restore the blunted Ang II contractions. In contrast, both DuP 753 and the structurally dissimilar SKF 108566 at a concentration of 100 nM showed complete recovery of Ang II responses within 2 hr of repeated washing. Surprisingly, repeated rinsing of rabbit aorta for up to 5 hr after incubation with 1 microM DuP 753 failed to restore responses to Ang II. In addition, Scatchard analysis of [125I] Ang II saturation binding experiments revealed a competitive and rapidly reversible nature of AT1 receptor antagonism for all the AT1 antagonists examined. Taken together, the results of this study provide evidence for a competitive and rapidly reversible binding interaction of structurally diverse non-peptide antagonists at the AT1 receptor.(ABSTRACT TRUNCATED AT 250 WORDS)
Panek
R L
RL
Department of Cardiovascular Therapeutics, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan, USA.
Lu
G H
GH
Overhiser
R W
RW
Major
T C
TC
Hodges
J C
JC
Taylor
D G
DG
D000803:Angiotensin I / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000378:metabolism; D002478:Cells, Cultured; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D011945:Receptors, Angiotensin / Q000378:metabolism*
D011945:Receptors, Angiotensin; D000803:Angiotensin I