The ability to assess the importance of VIP in different physiological processes is limited by the lack of specific potent antagonists. In the present study, we have adopted two different approaches used successfully with other peptides in an attempt to identify new VIP receptor antagonists. One involves the formation of pseudopeptides by insertion of reduced peptide bonds in the NH2-terminus from position 2 to 8 of VIP. The other methodology involves the formation of a COOH-terminal chimeric analogue by combining VIP(6-28) and PACAP(28-38). The ability of each of these peptides to function as an antagonist was compared with reported VIP antagonists. All of the peptides inhibited [125I]VIP binding to VIP receptors on guinea pig pancreatic acini. For the pseudopeptides the affinities were: [psi 3-4]VIP (0.2 microM) = 4 x [psi 4-5]VIP = 8 x [psi 8-9]VIP = 14 x [psi 6-7]VIP, [psi 2-3]VIP = 25 x [psi 5-6]VIP. Each nonpseudopeptide analogue also inhibited VIP binding with relative potencies of VIP(6-28)-PACAP(28-38) (1 microM) = 2.5 x [4-Cl-D-Phe6,Leu17]VIP, VIP(10-28), neurotensin(6-11)-VIP(7-28) = 6 x [Ac-Tyr1,D-Phe2]GRF. All pseudopeptides were agonists with relative potencies: [psi 3-4]VIP > [psi 6-7], [psi 4-5]VIP > [psi 5-6] > [psi 8- 9]VIP > [psi 2-3]VIP. The reported VIP receptor antagonist, neurotensin(6-11)-VIP(7-28), was also an agonist.(ABSTRACT TRUNCATED AT 250 WORDS)