D-pGlu 1, D-Phe 2, D-Trp 3,6]-LRF. A potent luteinizing hormone releasing factor antagonist [formula omitted] and inhibitor of ovulation in the rat

Abstract

The decapeptide D-pGlu-D-Phe-D-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2 was synthesized in gram quantities by solid phase techniques on a methyl benzhydrylamine resin. This peptide was purified by ion exchange and partition chromatographies. It was fully characterized by amino acid analysis, tlc in four systems, optical rotation and high pressure liquid chromatography (reversed phase) using a triethyl ammonium phosphate buffer. This peptide was found to inhibit ovulation by 100% in the rat at a subcutaneous dose of 250μg in corn oil when administered at noon on the day of proestrus. Furthermore, it was found to be long acting since 1.5mg blocked ovulation in 9 out of 10 rats when administered as early as 19 hrs before the afternoon of proestrus. [D-pGlu 1, D-Phe 2, D-Trp 3,6]-LRF reduces the amount of LH normally secreted in response to LRF by 50% at a molar ratio (IDR 50) of 3/1 ([antagonist])/[LRF]). Three other analogs, equally well-characterized and with the D-pGlu 1 modification, i.e. [D-pGlu 1, D-Phe 2, Pro 3, D-Trp 6]-LRF, [D-pGlu 1, D-Phe 2, Phe 3, D-Trp 6]-LRF and [D-pGlu 1, D-Phe 2,6, D-Trp 3]-LRF were found to be somewhat less potent ( IDR 50 = 150 1 , 60 1 , 5 1 ) respectively.