Synthesis and structure–activity investigation of novel vasopressin hypotensive peptide agonists

We report the solid phase synthesis and vasodepressor potencies of the novel hypotensive peptide [1(‐β‐mercapto‐β , β‐pentamethylene propionic acid)‐2‐ O ‐ethyl‐ D ‐tyrosine, 3‐arginine, 4‐valine] arginine vasopressin, d(CH 2 ) 5 [ D ‐Tyr(Et) 2 , Arg 3 , Val 4 ]AVP ( A ), its related Lys 3 ( B ), Tyr‐NH 2 9 ( C ), [Lys 3 , Tyr‐NH 2 9 ] ( D ) analogs and in a preliminary structure–activity study of positions 2–4 and 7–9, 24 analogs ( 1 – 24 ) of A – C . Peptides 1 – 6 , 9 – 14 have the following single substituents at positions 2, 3, 4, 8 and 9 in ( A ): 1 , D ‐Tyr(Me) 2 ; 2 , L ‐Tyr(Et) 2 ; 3 , Orn 3 ; 4 , N‐Me‐Arg 3 ; 5 , Glu 3 ; 6 , Arg 4 ; 9 , D ‐Arg 8 ; 10 , Eda 9 ; 11 , Arg‐NH 2 9 ; 12 , Ala‐NH 2 9 ; 13 , desGly 9 ; 14 , desGly‐NH 2 9 . Peptides 15 and 16 are analogs of B which possess the following single modifications: 15 , Arg‐NH 2 9 ; 16 , desGly 9 . Peptides 7 and 8 are analogs of ( C ) with the following single modification: 7 , Gln 4 ; 8 , Lys 8 . Peptides 17 – 24 are analogs of A possessing the following multiple modifications: 17 , [Sar 7 , Eda 9 ]; 18 , [Arg 7 , Eda 9 ]; 19 , [Arg 7 , Eda 9 ←Tyr 10 ]; 20 , [Arg 4 , Arg‐NH 2 9 ]; 21 , [Ile 4 , desGly 9 ]; 22 , [Arg 4 , desGly 9 ]; 23 , [Arg 7 , desGly 9 ]; 24 , [Arg 7 , Lys 8 , desGly 9 ]. All 24 new peptides were evaluated for agonistic and antagonistic activities in in vivo antidiuretic (V 2 ‐receptor), vasopressor (V 1a ‐receptor) and in in vitro (no Mg 2+ ) oxytocic (OT‐receptor) assays and like the parent peptides ( A – D ) (Chan et al . Br. J. Pharmacol. 1998; 125 : 803–811) were found to exhibit no or negligible activities in these assays. Vasodepressor potencies were determined in anesthetized male rats with baseline mean arterial blood pressure maintained at 110–120 mmHg. The effective dose (ED), in μg 100 g −1 i.v., required to produce a vasodepressor response of 5 cm 2 , area under the vasodepressor response curve (AUC) during the 5‐min period following the injection of the test peptide, was determined. Therefore, the EDs measure the relative vasodepressor potencies of the hypotensive peptides. The following ED values were obtained for A – D and for peptides 1 – 24 : A , 4.66; B , 5.75; C , 10.56; D , 11.60; 1 , ∼20; 2 , ∼30; 3 , 6.78; 4 , non‐detectable (ND); 5 , ND; 6 , ∼32; 7 , ND; 8 , 8.67; 9 , ND; 10 , 2.43; 11 , 3.54; 12 , 10.57; 13 , 4.81; 14 , ND; 15 , 4.47; 16 , 9.78; 17 , 5.72; 18 , 1.10; 19 , 1.05; 20 , 10.41; 21 , 9.13; 22 , ∼33; 23 , 3.01; 24 , 1.71. A is clearly the most potent of the four original hypotensive peptides A – D . These data provide insights to which modification of A enhance, retain or abolish hypotensive potencies. Six of the new hypotensive peptides are significantly more potent than A . These are peptides 10, 11, 18, 19, 23 and 24. Peptide 19, a radioiodinatable ligand, is ten times more potent than C or D . The Gln 4 modification of C and the N‐Me‐Arg 3 , Glu 3 , D ‐Arg 8 and desGly‐NH 2 9 modifications of A abolished hypotensive potency. By contrast, the Eda 9 , Arg‐NH 2 9 , [Sar 7 , Eda 9 ], [Arg 7 , Eda 9 ←Tyr 10 ], [Arg 7 , desGly 9 ], [Arg 7 , Lys 8 , desGly 9 ] modifications of A all led to enhancements of hypotensive potency. This initial structure–activity exploration provides useful clues to the design of (a) more potent vasodepressor peptides and (b) high affinity radioiodinatable ligands for the putative AVP vasodilating receptor. Some of the peptides here may be of value as pharmacological tools for studies on the complex cardiovascular actions of AVP and may lead to the development of a new class of anti‐hypertensive agents. Copyright © 1999 European Peptide Society and John Wiley & Sons, Ltd.