SYNTHESIS AND IDENTIFICATION OF 12 A-RING REDUCED 6-ALPHA-HYDROXYLATED AND 6-BETA-HYDROXYLATED COMPOUNDS DERIVED FROM 11-DEOXYCORTISOL, CORTICOSTERONE AND 11-DEHYDROCORTICOSTERONE

The synthesis and identification of 12 A-ring reduced 6 alpha-(and 6 b eta-)hydroxylated compounds derived from 11-deoxycortisol (S), cortico sterone (B) and 11-dehydrocorticosterone (A) are reported here. These steroids were prepared in two steps from the corresponding 6 6 alpha-( and 6 beta-)hydroxy-4-pregnene-3-ones. Selective reduction of the 4,5 double bond yielded 12 6 alpha-(and 6 beta)hydroxy-Scr(and 5 beta)preg nane-3,20-diones. Enzymatic reduction of these compounds with NADH and 3 alpha-hydroxysteroid dehydrogenase yielded the corresponding tetrah ydro steroids. The steroids were characterized by high performance liq uid chromatography (HPLC), gas chromatography mass spectrometry (GC an d GC/MS) and in part by H-1-NMR. 6 beta OH-THS and 6 beta OH-5 alpha T HS were identified by H-1-NMR. The structures of the two precursors, i .e. 6 beta OH-5 beta DHS and 6 beta OH-5 alpha DHS were confirmed by H -1-NMR using two-dimensional spectra. 6 alpha OH-THS was identified by comparing its HPLC, GC and MS data with those of the steroid obtained by enzymatic oxidation of the standard reference steroid 6 alpha OH-2 0 beta HHS to the corresponding 20-ketosteroid. The other steroids, e. g. GaOH-THB and 6 alpha OH-5 alpha THB were identified by using the pr oved sequence of elution of each of the epimer pairs on the normal pha se HPLC column (5 alpha < 5 beta), and by the reversed order of elutio n of the same epimer pair as the methoxime-trimethylsilyl ethers on th e GC column (5 alpha > 5 beta) and by the mass spectra, with the excep tion of 6 beta OH-THA.