Harold Levy

Cortisone metabolism in liver. II. Isolation of certain cortisone metabolites

Abstract 1. 1. A method of systematic analysis by paper chromatography of metabolites formed during cortisone perfusion through rat livers is presented. 2. 2. The isolation of allopregnan-3α, 17α,21-triol-11,20-dione, allopregnan-3β, 17α,21-triol-11,20-dione, allopregnan-3α, 11β,17α,21-tetrol-20-one, allopregnan-3β, 11β,17α, 21-tetrol-20-one, Δ 4 -pregnen-11β,17α,21-triol-3,20-dione, Δ 4 -pregnen-17α,20β,21-triol-3, 11-dione, and adrenosterone from cortisone perfusates is described. 3. 3. The relationship of these findings to an understanding of pathways of corticosteroid metabolism in liver is briefly discussed.

The isolation of 18-hydroxy-3-oxo-androst-4-ene-17β-carboxylic acid-20, 18-lactone, its 11β-hydroxy analog and other substances from bovine adrenal perfusions of deoxycorttcosterone

Abstract The nature of the substances formed from deoxycorticosterone in bovine adrenal perfusion has been reinvestlgated. In addition to corticosterone, the following substances were isolated in crystalline form and identified: 11β,21-dihydroxy-5α-pregnan-3,20-dione; 3β, 11β,21-trihydroxy-5α-pregnan-20-one; 11β,20β,21-trihydroxypregn-4-en-3-one; 11β-hydroxy-3-oxoandrost-4-en-17β-carboxylic acid; 18-hydroxy-3-oxoandrost-4-en-17β-carboxylic acid-20,18-lactone; 11β, 18-dihydroxy-3-oxoandrost-4-en-17β-carboxylic acid 20,18-lactone; “dimer” of 18-hydroxy-deoxycorticosterone; and 19-hydroxy-deoxycorticosterone. Sixteen other metabolites were crystallized but not identified.

The inhibition of 11β-hydroxylation of deoxycorticosterone by metopirone in bovine adrenal perfusions

Abstract Deoxycorticosterone (DOC), 125 mg/l, 3.8 × 10−4 M, in bovine blood was perfused, through bovine adrenal glands in the presence of a varying amount of Metopirone (100-3.15 mg/l, 4.4−0.14 × 10−4M) and for a varying number of cycles (5–35) to determine whether 11β-hydroxylation to corticosterone would be inhibited by Metopirone in this system and whether DOC would be shunted into other metabolic pathways. With Metopirone at 100-25 mg/l, the inhibition of 11β-hydroxylation was essentially complete in the standard 5-cycle perfusion and was almost complete even after 30 cycles. At 3.15 mg/l,the inhibition was still about 65% effective after 5 cycles and about 30% effective after 22 cycles. The general metabolism of DOC in other directions was slowed. With Metopirone at 100 mg/l, 84% of the DOC was recovered after 5 cycles. At 3.15 mg/l, the recovery was still about 40%. However, 13 metabolites other than corticosterone were isolated from the various perfusates, all in conversions of 3% or less. Five substances were identified. Of these, 6β-hydroxy-DOC was formed even within 5 cycles with Metopirone at 100 mg/l. Neither this substance nor the 8 unidentified compounds have been isolated from non-inhibited perfusions.

The monohydroxylation of 17β-estradiol at the 15α-, 16α-, and 16β-positions by bovine adrenal perfusion

Abstract 17β-Estradiol-4-14C was perfused through bovine adrenal glands. The bulk (65%) of the estradiol was recovered, but several metabolites, all in low rates of conversion, were isolated. Four of these were identified as esterone, estriol, 16β-epiestriol and 15-αhydroxy-17β- estradiol.

The inhibition by metopirone of 11β- and 19-hydroxylations of 11-deoxycortisol in bovine adrenal perfusion

Abstract When 11-deoxycortisol, 3.6 × 10−4M, was perfused through cow adrenals for 5 cycles, cortisol (55.0% conversion) and 19-hydroxy-11-deoxycortisol (2.1%) were isolated. No precursor was recovered. When this perfusion was repeated in the presence of Metopirone, 1.1 × 10−4M, cortisol (0.2%) was obtained and deoxycortisol (63.5%) was recovered. An extension of the latter perfusion to 29 cycles led to the isolation of 17,21-dihydroxy-5α-pregnane-3,20-dione (3.5%), deoxycortisol (31.5%), cortisol (2.5%), 6β-hydroxy-11-deoxycortisol (2.1%) and 19-hydroxy-11-deoxycortisol (0.8%). Four other metabolites were isolated from inhibited perfusions but not identified. Conclusions: Metopirone inhibited 11β- and, probably, 19-hydroxylation but permitted 6β-hydroxylation.

The selective inhibition by metopirone of 11β-hydroxylation in bovine adrenal perfusion of 17-hydroxyprogesterone (1)

Abstract When progesterone-4-14C, 2.2–2.4 × 10−4M, was perfused through cow adrenal glands for 10 cycles in the presence of Metopirone, 1.1 × 10−4M, the substances isolated included 11-deoxycortisol (18.5% and 22.6% conversion) and deoxycorticosterone (2% and 4.7%) but not cortisol or corticosterone. Without Metopirone, the first two metabolites would not be end-products and the conversions into the last two metabolites would be about 18% and 11%. Therefore, Metopirone inhibited 11β-hydroxylation but permitted 17α- and 2l-hydroxylations. When the perfusion was extended to 20-cycles, the isolated compounds included 11-deoxycortisol (41.1%) and cortisol (2.8%) but not deoxycorticosterone or corticosterone. Thus, the inhibition of 11β-hydroxylation of deoxycortisol but not of deoxycorticosterone was overcome slightly under continued cycling.

Conversion of 17-hydroxyprogesterone into 3α,17-dihydroxypregn-4-en-20-one, 17,20β-dihydroxypregn-4-en-3-one and other substances by bovine adrenal perfusion (1)

Abstract 3α,17-Dihydroxypregn-4-en-20-one, 17,20β-dihydroxypregn-4-en-3-one, 17-hydroxy-5α-pregnane-3,20-dione and 3β,17-dihydroxy-5α-pregnan-20-one were isolated from bovine adrenal perfusates of 17-hydroxyprogesterone.

The inhibition by metopirone of 11β- and 19-hydroxylations of androst-4-ene-3,17-dione in bovine adrenal perfusion (1)

Abstract Androst-4-en-3,17-dione-4-14C, 350 μM, was perfused through cow adrenals for 5–6 and 30 cycles in the absence of Metopirone and in its presence at 110 μM and 55 μM. Hydroxylations at 11β and 19, the chief reactions in the unmodified perfusion, were both completely inhibited (or virtually so) by Metopirone at the higher concentration and were reduced to about 6% of the control values at the lower concentration, even after 30 cycles. Reductions of the C4-double bond and the C3-carbonyl group were not inhibited. Oxidation to 6-oxoandrostenedione occured to a small extent in both types of perfusion but no conclusion about the influence of Metopirone on this reaction can be drawn. Nonradioactive 3β, 17-dihydroxypregn-5-en-20-one was isolated from one of the inhibited perfusions.