Urinary Alpha-Ketolic Steroid Metabolites of Cortical Hormones Administered to Subjects With Adrenal Cortical Insufficiency 1

Abstract

The analysis of urinary excretory products following the administration of known compounds is one of the classical methods for the study of their metabolism. The developments in paper chromatographic (1) and micro-analytic (2) technics and the use of glucuronidase hydrolysis (3) have considerably decreased the difficulties of investigating steroid metabolism by this method. Using these newer technics, we have studied the urinary excretion pattern of the major alpha-ketolic steroids after the administration of the six "active" adrenal cortical steroids to individuals with severe adrenal cortical insufficiency. We have found, as has been demonstrated for other steroids (4), that the major alpha-ketolic metabolites result from the reduction in ring A of the steroid nucleus to the 3-alpha-hydroxy pregnane derivatives (the socalled "tetrahydro" form) and that the hydroxyl and ketone groups on carbon atom 11 are interconvertible for corticosterone and 1 1-dehydrocorticosterone as well as for hydrocortisone and cortisone. METHODS I. TerminologySince steroid terminology is varied and the proper chemical names consume considerable space, the terms and abbreviations more commonly used in physiological research will be employed in this paper. Definitions and abbreviations are listed below.Alpha-ketolic steroids: Pregnane derivatives having 20-keto and 21-hydroxy substituents (i.e., alpha-ketol group). A4-pregnene-21-ol-3, 11, 20-trione.DOC; li-desoxycorticosterone; A'-pregnene-21-ol-3, 20-dione. These are listed in order of decreasing polarity. The acetate is indicated by the abbreviation ac.Alpha-ketolic steroid metabolites (AKSM): Steroids with the alpha-ketolic side chain found in the urine. These are detectable by reaction of the alpha-ketol side chain with alkaline blue tetrazolium to produce a blue color. The unmodified abbreviation H4 indicates reduction in ring A to form the 3a normal (i.e., pregnane) reduction product. In addition to the alpha-ketolic cortical hormones, the major metabolites are: H4F; tetrahydro-hydrocortisone; pregnane-3a, 11P, 17a, 21-tetrol-20-one. H4E; tetrahydro-cortisone; pregnane-3a, 17a, 21-triol-11, 20-dione. H4S; tetrahydro-11-desoxy-17-hydroxycorticosterone; pregnane-3a, 17a, 21-triol-20-one. H,B; tetrahydro-corticosterone; pregnane-3a, Ilfi, 21-triol-20-one. H4A; tetrahydro-dehydrocorticosterone; pregnane-3a, 21-diol-11, 20-dione. H4DOC; tetrahydro-l1-desoxycorticosterone; pregnane-3ca, 21-diol-20-one. Polarity: In the chromatograms, the most polar steroids (i.e., those with the most hydroxyl and keto substituents) appear in the slowest moving fraction, X, and the least polar in the fastest moving fraction, Z. (See III, B Paper chromatographic and micro-analytic technics.) 285

The Effect of Acth and Pathological Increases in Adrenal Cortical Function on Urinary Alphaketolic Steroid Metabolites 1

“…after ACTH. It is evident from this and previous studies (1) as well as the position and relatively small absorption at 245 m,u (not shown) in these regions that the twin alphaketolic peaks in the first portion of fraction X in the other subjects are probably formed by H4F and H4E. The increased excretion of hydrocortisone (F) after administration of ACTH is also indicated.…”

“…Methods employed and their limitations have been presented previously (1). Terminology has been previously discussed (1).…”

“…After development of the chromatogram, the absorption of ultra-violet light of the paper strip at 245 mn was determined directly in a Beckman DU spectrophotometer ' This investigation was supported in part by research grant H-922 from the National Heart Institute, of the Heard, Sobel, and Venning (4) after acid hydrolysis (Ac. R-L) and after glucuronidase followed by acid hydrolysis (G'ase R-L) as previously described (1). The total neutral 17-ketosteroids (17-KS) were determined by a modification of the method of Holtorff and Koch (5).…”

“…The most polar (X), "mid-polar" (Y), and least polar (Z) fractions are charted. The percentage figures in each fraction indicate the proportion of the 24-hour urine represented by the amount of extract used tration of F or E (1,(8)(9)(10)(11) and the last four are those found in large amounts after administration of compounds B or A (1,12). Cortisone (E), although indicated (Table I), was not positively identified after administration of ACTH but has been positively identified by others (Table III).…”

“…The ratio of the estimated quantities of the various metabolites have been examined and compared to the ratios found after oral administration of F or E and B or A in the previously reported experiments (1). It is recognized that dosage and route of administration may influence the absolute values and the ratios.…”