Synthesis and gas chromatographic-mass spectrometric analysis of reduced compounds derived from 6α- and 6β-hydroxy-11-deoxycorticosterone

Bournot, Paulette; Ramirez, Leyla C.; Pitoizet, Nicole; Maume, B.F.; Padieu, P

doi:10.1016/0039-128x(89)90151-7

Cited by 8 publications

(2 citation statements)

References 26 publications

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“…As the steroid was not 11b-hydroxylated, the most likely position would be 6a as hydroxylation at this position is common for ring-A saturated corticosteroids. The spectrum is in fact identical to the reported spectrum of 6a-hydroxytetrahydroDOC (Bournot et al, 1989), and we propose that the isolated steroids are epimers of 6-hydroxytetrahydroDOC. It is not possible to distinguish the 6a from the 6b form by MS because of an unavailability of reference standards.…”

Section: Resultssupporting

confidence: 84%

“…After HPLC separation a total of nine fractions were collected and analyzed by GC-MS. Steroids giving the mass spectrum of tetrahydroDOC(Bournot et al, 1989) were found in HPLC fraction 6 (Fig2). The steroid with a retention time of 18.36 min is 3a,21-dihydroxy-5a-pregnan-20-one (3a,5aTHDOC) based on its having a retention time identical to the authentic standard steroid.…”

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confidence: 99%

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Gas Chromatography/Mass Spectrometry Characterization of Corticosteroid Metabolism in Human Immortalized Keratinocytes

Slominski

Wortsman

Foecking

et al. 2002

Journal of Investigative Dermatology

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To continue our studies on the cutaneous expression of a proopiomelanocortin/corticotropin-releasing hormone system, we investigated whether this is accompanied by adrenal-type enzymatic activity. Immortalized cultured human keratinocytes were incubated with radiolabeled corticosteroids. Analysis by thin-layer chromatography showed rapid transformation of both progesterone and deoxycorticosterone; one of the progesterone metabolites migrated at the same rate as deoxycorticosterone. Gas chromatography/mass spectrometry further identified as major species of deoxycorticosterone metabolites 3beta,6alpha,21-trihydroxy-5alpha-pregnan-20-one, 3alpha,6alpha,21-trihydroxy-5alpha-pregnan-20-one, and 3alpha5alpha- and 3beta5alpha-tetrahydrodeoxycorticosterone. Minor metabolites were 3alpha,21-dihydroxy-5-pregnen-20- one (3alphaDelta5-21-OHpregnenolone), 3beta,21-dihydroxy-5-pregnen-20-one (3betaDelta5-21-OHpregnenolone), 3alpha,21-dihydroxy-4-pregnen-20-one (3alphaDelta4-21-OHpregnenolone), 6-hydroxy-dihydrodeoxycorticosterone, and two 5-dihydrodeoxycorticosterone species. Thus, in addition to sex steroids keratinocytes also actively metabolize corticosteroids along similar enzymatic pathways. The surprising detection of 3alphaDelta5-21-OHpregnenolone and 3 betaDelta5-21-OHpregnenolone, indicating Delta4-ketosteroids to Delta5-hydroxysteroids conversion, provides strong evidence for the occurrence, at least in human keratinocytes, of isomerase activity that allows the reaction to proceed in reverse of its usual direction. As skin expresses 3alpha/beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerases, cutaneous reactions catalyzed by these enzymes must be reversible. In conclusion, besides elements of the corticotropin-releasing hormone/proopiomelanocortin system human keratinocytes show high levels of corticosteroid metabolizing activity. Moreover, the wide array of steroid products generated from a single substrate indicates serial progressive conversion involving 5alpha-reductase, 6alpha-hydroxylase, 3alpha/beta-hydroxysteroid dehydrogenase, and reverse Delta4minus signDelta5 isomerase enzymes. As distinct from the adrenal cortex, production of A, B, Aldo, 18OHdeoxycorticosterone, or F in keratinocytes was absent or below limits of detectability.

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confidence: 84%

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confidence: 99%