G. Kraan, J. Hartstra, B. Wolthers
Jun 1, 1994
Citations
1
Influential Citations
3
Citations
Journal
The Journal of Steroid Biochemistry and Molecular Biology
Abstract
Abstract The synthesis and identification of 12 A-ring reduced 6α-(and 6β-)hydroxylated compounds derived from 11-deoxycortisol (S), corticosterone (B) and 11-dehydrocorticosterone (A) are reported here. These steroids were prepared in two steps from the corresponding 6 6α-(and 6β-)hydroxy-4-pregnene-3-ones. Selective reduction of the 4,5 double bond yielded 12 6α-(and 6β)hydroxy-5α-(and 5β)pregnane-3,20-diones. Enzymatic reduction of these compounds with NADH and 3α-hydroxysteroid dehydrogenase yielded the corresponding tetrahydro steroids. The steroids were characterized by high performance liquid chromatography (HPLC), gas chromatography mass spectrometry (GC and GC/MS) and in part by 1 H-NMR. 6βOH-THS and 6βOH-5αTHS were identified by 1 H-NMR. The structures of the two precursors, i.e. 6βOH-5βDHS and 6βOH-5αDHS were confirmed by 1 H-NMR using two-dimensional spectra. 6αOH-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 6αOH-20βHHS to the corresponding 20-ketosteroid. The other steroids, e.g. 6αOH-THB and 6αOH-5αTHB were identified by using the proved sequence of elution of each of the epimer pairs on the normal phase HPLC column (5 α β ), and by the reversed order of elution of the same epimer pair as the methoxime-trimethylsilyl ethers on the GC column (5α > 5β) and by the mass spectra, with the exception of 6βOH-THA.