Synthesis of 3-hydroxyestra-1,3,5(10)-trien-17-one and 3,17 beta-dihydroxyestra-1,3,5(10)-triene 6 alpha-N-(epsilon-biotinyl) caproamide, tracer substances for developing immunoassays for estrone and estradiol.
We describe the synthesis of 3-hydroxyestra-1,3,5(10)-trien-17-one 6 alpha-N-(epsilon-biotinyl)caproamide and 3,17 beta-dihydroxyestra-1,3,5(10)-triene 6 alpha-N-(epsilon-biotinyl) caproamide from 3-hydroxyestra-1,3,5(10)-trien-17-one and 3,17 beta-dihydroxyestra-1,3,5(10)-triene, via the 6-keto estrogenic derivatives. The reductive amination of these compounds is an effective step toward an epimeric mixture of the respective amines, which are easily biotinylated by use of N-(epsilon-biotinylcaproyl)-N- hydroxysuccinimide ester. The 6 alpha-epimers could be isolated from the alpha/beta-composition by application of isocratic HPLC, and overall yields were about 20% for the epimeric end products. The structures of the stereoisomers could clearly be assigned through 1H NMR studies. The ratios of the respective isomers obtained from the reductive amination were found to be 3(alpha):2(beta). The biotinylated estrogens can be used as tracers in a novel immunoassay concept for the determination of these analytes in human serum. Ring position 6 was selected for derivatization because of its distance from the functionalized positions 3 and 17 and, therefore, of a negligible alteration of the tracer's structure in comparison to underivatized estrone or estradiol.
Luppa
P
P
Institute for Clinical Chemistry, Technical University Munich, Germany.
Birkmayer
C
C
Hauptmann
H
H
D001667:Binding, Competitive; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D004958:Estradiol / Q000031:analogs & derivatives* / Q000032:analysis* / Q000138:chemical synthesis / Q000276:immunology; D004970:Estrone / Q000031:analogs & derivatives* / Q000032:analysis* / Q000138:chemical synthesis / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D009682:Magnetic Resonance Spectroscopy; D013237:Stereoisomerism
C088376:3,17-dihydroxyestra-1,3,5(10)-triene 6-N-(epsilon-biotinyl)caproamide; C088353:3-hydroxyestra-1,3,5(10)-trien-17-one 6-N-(epsilon-biotinyl)caproamide; D004970:Estrone; D004958:Estradiol; D001710:Biotin