Stereoselective distribution of the teratogenic thalidomide analogue EM12 in the early embryo of marmoset monkey, Wistar rat and NMRI mouse

Abstract

 Thalidomide administration during early gestation results in specific and dramatic limb defects in primates, but not in laboratory rodents such as the rat and mouse. The thalidomide analogue EM12 [2-(2, 6- dioxopiperidine-3-yl)-phthalimidine] was used in the present study because this compound is metabolically more stable and teratogenically more potent than thalidomide in the monkey. We have administered the pure enantiomers, since we have previously shown that S-EM12 proved to be much more teratogenic in the monkey than R-EM12. In maternal plasma, placenta and embryo of the pregnant marmoset monkey (Callithrix jacchus) and Wistar rat, the concentrations were investigated of the enantiomers and their metabolites after administration of R- and S-EM12. With whole body autoradiography the distribution in the embryo, including the target tissue, the embryonic limb bud was examined in the NMRI mouse and marmoset monkey. Our investigations showed that both the R- and the S-enantiomers were transferred to the embryo during organogenesis [monkey, gestation day (GD) 61; rat, GD 12; mouse, GD 10]. The gestation period chosen was toward the end of the thalidomide-sensitive stage, but yielded sufficient gestational material for analysis. Considerable amounts of the enantiomers were produced via racemization of the administered pure enantiomers and were present in maternal plasma as well as in placenta and embryo. In the monkey, the racemization were stereoselective: the S-enantiomer was eliminated more slowly in the monkey than the R-enantiomer, possibly because of stereospecific binding and metabolism. In the plasma and embryo of both rat and monkey, the metabolites were detected in considerably lower concentrations than EM12, emphasizing the importance of the parent drug in regard to the teratogenic effect. The whole-body autoradiography in marmoset and mouse showed high radioactivity in the embryonic CNS, the branchial apparatus and in the limb buds. The S-enantiomer of EM12 was more strongly concentrated than the R-enantiomer in these areas. In the limb buds, the highest concentrations of radioactivity were observed in the periphery, sometimes at the very tip of the buds. Accumulation of radioactivity in limb buds and neural epithelium relative to other areas of the embryo was much more pronounced in the monkey than in the mouse. Future studies must demonstrate if this accumulation has implications for the mechanism of thalidomide teratogenesis in primate species.