cortodoxone and 11-dehydrocorticosterone

A recombinant expression vector containing Japanese eel (Anguilla japonica) testis cytochrome P450(11 beta) (11beta-hydroxylase) cDNA was introduced into COS-1 cells. Enzymatic activity of the expressed P450(11 beta) for corticosteroid synthesis was analysed by incubating transfected cells with 14C-labelled 11-deoxycorticosterone or 3H-labelled deoxycortisol as substrates. Thin layer chromatography of incubation medium revealed that a high percentage of 11-deoxycorticosterone was converted into corticosterone and 11-dehydrocorticosterone but no aldosterone was detected. Similarly, deoxycortisol was converted into cortisol and cortisone. These results show that eel P450(11beta) does not possess significant aldosterone synthesizing activity. Northern blot analysis detected a 1.8 kb transcript of P450(11beta) using RNA extracted from interrenals of untreated Japanese eel but no hybridization signal was apparent using RNA extracted from brain, spleen, heart, muscle or testis. Immunohistochemistry using an antiserum against P450(11beta) also revealed strong immunostaining in interrenal cells.

Topics: Aldosterone; Anguilla; Animals; Corticosterone; Cortisone; Cortodoxone; COS Cells; Desoxycorticosterone; Gene Expression; Hydrocortisone; Interrenal Gland; Male; Recombinant Proteins; Steroid 11-beta-Hydroxylase; Testis; Transfection

The synthesis and identification of 12 A-ring reduced 6 alpha-(and 6 beta-)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 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-5 alpha-(and 5 beta)pregnane-3,20-diones. Enzymatic reduction of these compounds with NADH and 3 alpha-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 1H-NMR. 6 beta OH-THS and 6 beta OH-5 alpha THS were identified by 1H-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 1H-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-20 beta HHS to the corresponding 20-ketosteroid. The other steroids, e.g. 6 alpha OH-THB and 6 alpha OH-5 alpha THB were identified by using the proved sequence of elution of each of the epimer pairs on the normal phase HPLC column (5 alpha < 5 beta), and by the reversed order of elution of the same epimer pair as the methoxime-trimethylsilyl ethers on the GC column (5 alpha > 5 beta) and by the mass spectra, with the exception of 6 beta OH-THA.

Topics: 3-alpha-Hydroxysteroid Dehydrogenase (B-Specific); 3-Hydroxysteroid Dehydrogenases; Chromatography, High Pressure Liquid; Corticosterone; Cortodoxone; Gas Chromatography-Mass Spectrometry; Hydroxylation; Magnetic Resonance Spectroscopy; NAD; Oxidation-Reduction; Steroids

Mineralocorticoid receptors (MR) are protected from the effects of endogenous glucocorticoids (GC) in mineralocorticoid (MC) target tissues such as the kidney and the parotid gland. This protection is thought to be provided by 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD). 11 beta-OHSD metabolizes cortisol (in humans) and corticosterone (B) (in the rat) to cortisone and 11-dehydro-B, their respective inactive dehydro products. We have previously shown that the antinatriuretic actions of the MC deoxycorticosterone (DOC) are amplified in carbenoxolone (CBX) treated adrenalectomized (ADX) rats. CBX is believed to inhibit 11 beta-OHSD activity; DOC, however, is not a substrate for this enzyme. We now report on 11-desoxycortisol (11-desoxy-F) and 2 alpha-methylcortisone, substances which possess no intrinsic antinatriuretic activity, are not metabolized by 11 beta-OHSD and yet cause Na+ retention in CBX-treated ADX rats. Given that none of the above steroids are substrates for 11 beta-OHSD it is unlikely that the inhibition of this enzyme is involved in the unmasking of the Na+ retention observed when these substances are given to CBX-treated animals. These results provide further evidence for an additional protective mechanism, that protects MR from the inappropriate binding of excessive amounts of endogenous MCs.

Topics: 11-beta-Hydroxysteroid Dehydrogenases; Aldosterone; Animals; Carbenoxolone; Corticosterone; Cortisone; Cortodoxone; Hydroxysteroid Dehydrogenases; Male; Mineralocorticoids; Natriuresis; Potassium; Rats; Rats, Sprague-Dawley

This report describes the synthesis of 6 alpha, 17,21- and 6 beta, 17,21-trihydroxypregn-4-ene-3,20-dione, 6 alpha, 7,21- and 6 beta, 11 beta, 21-trihydroxypregn-4-ene-3,20-dione, and--for the first time--that of 6 alpha, 21- and 6 beta, 21-dihydroxypregn-4-ene-3,11,20-trione. The former four compounds were prepared by 6-hydroxylation of 17,21-trihydroxypregn-4-ene-3,20-dione and 11 beta, 21-dihydroxypregn-4-ene-3,20-dione, respectively. This was achieved by autoxidation or by oxidation with 3-chloroperbenzoic acid, of the 3-methoxy-pregna-3,5-dienes of the latter two steroids. The yield of the 6 beta-hydroxylated steroids, but not of their corresponding 6 alpha-epimers, was higher using autoxidation than the peracid. The two 6-hydroxylated pregnenetriones were prepared from 6 alpha, 21-diacetoxy-11 beta-hydroxypregn-4-ene-3,20-dione and 6 beta, 21-diacetoxy-11 beta-hydroxypregn-4-ene-3,20-dione, respectively, by oxidation with pyridinium chlorochromate. The above-mentioned six steroids were identified and characterized by nuclear magnetic resonance, infrared, ultraviolet, high performance liquid chromatography, gas chromatography, and mass spectrometry.

Topics: Chlorobenzoates; Chromatography, Gas; Chromatography, High Pressure Liquid; Corticosterone; Cortodoxone; Gas Chromatography-Mass Spectrometry; Hydroxylation; Magnetic Resonance Spectroscopy; Mass Spectrometry; Oxidation-Reduction; Spectrophotometry, Infrared; Spectrophotometry, Ultraviolet

Corticosterone (B), aldosterone (Aldo), 11-deoxycorticosterone (DOC), 11-dehydrocorticosterone (A), 18-hydroxycorticosterone (18 OH B) and cortisol (F) are identified after incubation of interrenal of Bufo bufo formosus with radioactive progesterone. Yields of radioactive B and Aldo are larger than those of radioactive DOC, A and 18 OH B; yield of radioactive F is the smallest one.

Topics: 18-Hydroxycorticosterone; Adrenal Glands; Aldosterone; Animals; Bufo bufo; Corticosterone; Cortodoxone; Female; Hydrocortisone; In Vitro Techniques; Interrenal Gland; Male

Topics: Adrenocorticotropic Hormone; Animals; Chromatography, Thin Layer; Corticosterone; Cortodoxone; Female; Fishes; Hagfishes; Hydrocortisone; Hydroxycorticosteroids; Hydroxyprogesterones; Lampreys; Male; Testosterone

The urinary excretion of 17-hydroxycorticosteroids and the relations between the glucocorticoids in urine and their precursors as well as between 17-hydroxycorticoids and 17-hydroxycorticoids and 17-dehydroxycorticosteroids was measured in two subjects before and after 8 days flight in spaceship "Soyuz-22". During a readaptation period after the space flight activation of the glucocorticoid function of adrenals was observed which was accompanied by signs of stress and relative deficiency of 11-hydroxylation in glucocorticoid synthesis. The assumptions on possible causes of observed changes are discussed.

Topics: Adrenal Cortex Hormones; Aerospace Medicine; Corticosterone; Cortisone; Cortodoxone; Humans; Hydrocortisone; Male; Space Flight; Tetrahydrocortisol; Tetrahydrocortisone

A high-performance liquid chromatographic method is described for the determination of seven steroids in adrenocortical extracts showing a delta 4-3-ketonic conjugated system. The seven steroids (cortisol, cortisone, 11-dehydrocorticosterone, corticosterone, 11-deoxycortisol, aldosterone and 11-deoxycorticosterone) were separated with a chloroform-methanol gradient on a 5-micron silica column and with a water-acetonitrile gradient on a 10-micron RP-8 column. Effluents were monitored by UV absorption at 242 nm. Quantitative analysis was performed by comparing peak areas, which are proportional to the amounts of the individual substances (external standard method). The method is rapid, sensitive, easy to perform and reproducible.

Topics: Adrenal Cortex Hormones; Aldosterone; Chromatography, High Pressure Liquid; Corticosterone; Cortisone; Cortodoxone; Desoxycorticosterone; Hydrocortisone; Ketosteroids; Tissue Extracts

Topics: Adrenal Cortex; Adrenal Cortex Hormones; Adrenocorticotropic Hormone; Body Fluids; Corticosterone; Cortodoxone; Pregnanes; Urine

Topics: Acetates; Adrenal Cortex; Adrenal Cortex Hormones; Cardiovascular System; Corticosterone; Cortodoxone; Desoxycorticosterone; Desoxycorticosterone Acetate; Hypertension