Collect. Czech. Chem. Commun. 1996, 61, 799-807

Radioimmunological and Chromatographic Properties of Tyrosine Methyl Ester Conjugates with Stereoisomeric Steroid Carboxy Derivatives

Oldřich Lapčíka, Richard Hampla, Martin Hilla, Luboslav Stárkaa, Alexander Kasalb, Vladimír Pouzarb and Zdeněk Putzc

a Institute of Endocrinology, 116 94 Prague 1, Czech Republic
b Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, 166 10 Prague 6, Czech Republic
c Institute of Endocrinology, 034 91 Ľubochňa, Slovak Republic


Pure 3Z (syn) and 3E (anti) stereoisomers of testosterone 3-[O-(2-carboxyethyl)]oxime were synthesized, separated by HPLC or TLC, and used for preparation of tyrosine methyl ester (TME) conjugates by using mixed anhydride or carbodiimide-N-hydroxysuccinimide methods. While the latter method provided more than 96% of product with retained configuration, the mixed anhydride method yielded a mixture containing 26-40% of the opposite stereoisomer. The stereoisomers were used as model compounds, to which the other steroid TMEs and the corresponding radioiodinated products could be aligned according to their chromatographic properties. The TME conjugates of 3-(O-carboxymethyl)oximes of seven 4-en-3-oxo steroids were further prepared by carbodiimide-N-hydroxysuccinimide method. With exception of cortisol, the stereoisomeric (Z and E) radioiodinated TME conjugates could be separated by TLC. In addition, the conjugates with TME and consequently radioiodinated tracers were synthesized from hemisuccinates of cortisol and its 11α-isomer, via 11β- and 11α-hydroxy group. The radioiodinated conjugates were tested as radioligands with rabbit polyclonal antisera raised by using position-homologous conjugates of the respective steroid carboxy derivatives with bovine serum albumin as immunogens. With the exception of 11-deoxycorticosterone, the stereoisomeric Z and E radioiodinated TMEs did not differ in their binding properties. In the case of isomeric cortisol tracers conjugated through position 11 the antisera recognized only the sterically homologous radioligands, but the specificity of the system was poor.

Keywords: Steroid carboxymethyl derivatives; Tyrosine methyl esters; Radioiodination; Radioimmunoassay.