The metabolism of 20-hydroxyecdysone in mice: Relevance to pharmacological effects and gene switch applications of ecdysteroids

Ecdysteroids exert many pharmacological effects in mammals (including humans), most of which appear beneficial, but their mechanism of action is far from understood. Whether they act directly and/or after the formation of metabolites is still an open question. The need to investigate this question has gained extra impetus because of the recent development of ecdysteroid-based gene-therapy systems for mammals.In order to investigate the metabolic fate of ecdysteroids in mice, [1α,2α-3H]20-hydroxyecdysone was prepared and injected intraperitoneally to mice. Their excretory products (urine + faeces) were collected and the different tritiated metabolites were isolated and identified. The pattern of ecdysteroid metabolites is very complex, but no conjugates were found, in contrast to the classical fate of the (less polar) endogenous vertebrate steroid hormones. Primary reactions involve dehydroxylation at C-14 and side-chain cleavage between C-20 and C-22, thereby yielding 14-deoxy-20-hydroxyecdysone, poststerone and 14-deoxypoststerone. These metabolites then undergo several reactions of reduction involving, in particular, the 6-keto-group.A novel major metabolite has been identified as 2β,3β,6α,22R,25-pentahydroxy-5β-cholest-8(14)-ene. The formation of this and the other major metabolites is discussed in relation to the various effects of ecdysteroids already demonstrated on vertebrates.

► Ecdysteroids display a wide range of pharmacological effects on mammals, but their targets have not been identified. They undergo extensive metabolism in mammals. ► 20-hydroxyecdysone labelled on the steroid nucleus has been prepared and administered to mice. ► The major metabolites have been isolated and fully identified by MS and NMR and comparison with synthetic molecules. ► This opens new ways of thinking for identifying the mechanism of action of ecdysteroids on mammals.