Dehydroepiandrosterone isolation

An improved route has been described for the degradation of cholesterol to dehydroepiandrosterone,68,69 in which cholesterol is first acetylated and then bromi-nated to give the 5a,6/8-dibromide. This protected cholestane is then isomerized to the 5/3,6a-dibromide prior to chromium trioxide oxidation and zinc debromination. The yield of dehydroepiandrosterone (isolated from the reaction mixture as a 5a,6/3-dibromide) is 20%. 

Papaverine dehydroepiandrosterone-3-monophosphate (151) had a much more rapid coronary vasodilator action on the isolated cat heart than papaverine the effect of papaverine was, however, of longer duration. Both compounds inhibited histamine bronchospasms. The spasmolytic effect of papaverine dehydroepiandrosterone-3-mono-phosphate on histamine-induced contractions of the small intestine of guinea pigs was twice as strong as that of papaverine but less toxic than papaverine. 

Formation of sulfates [328] is relatively reversible. This may be due to the fact that the metabolic clearance rate [318] of sulfates is relatively low [389] and their renal excretion inefficient [4]. The conversion of dehydroepiandrosterone sulfate in vivo to androsterone and etiocholanolone glucuronide was demonstrated by Lieberman and co-workers, who administered isotopically labeled dehydroepiandrosterone sulfate and isolated labeled androsterone and etiocholanolone from glucuronicase-hydrolyzed urine [303]. Since in another study by Lieberman [174], the transconjugation from dehydroepiandrosterone sulfate to dehydroepiandrosterone glucuronide without free dehydroepiandrosterone intermediate was declared to be improbable, in vivo equilibrium between dehydroepiandrosterone sulfate and dehydroepiandrosterone seems probable [303]. 

According to the classical view of metabolism the hormones are synthesized as free steroids in endocrine tissues and prepared for excretion in urine by peripheral metabolism and conjugation. This view had to be modified upon the isolation of dehydroepiandrosterone sulfate from adrenal tumor [307]. Thus dehydroepiandrosterone sulfate, a steroid conjugate, was shown to be secreted by the adrenal tissue. Isotopic methods also pointed in the same direction. Lieberman et al. [304], using... 

Simmer, H. H., Frankland, M. V., Greipel, M., and Zlotnik, G., Isolation and identification of unconjugated dehydroepiandrosterone and 16a-hydroxydehydro-epiandrosterone from pooled cord blood. Steroids 11, 13-26 (1968). 

Androgens, the male sex hormones, proved far more elusive that either the estrogens and progestins since they occur at much lower concentrations in biological fluids. The bioassay used to track the isolation in this case comprised the capon unit . This was the amount of extract that produced a 20% increase in the surface of a rooster s comb. The 15 mg of pure crystalline testosterone isolated in 1931 came from about 15 0001 of urine. The structural investigations of this series relied on the then newly discovered side chain oxidations of cholestanol (13-1) (Scheme 1.13). This method in essence comprised fairly drastic oxidation of reduced cholesterols of known stereochemistry at the A-B junction to afford in fairly low yield products in which the side chain at Cn had been consumed to leave behind a carbonyl group. One of these products proved to be identical with androsterone (13-2). That compound had in turn been obtained from a sequence of reactions starting from dehydroepiandrosterone (13-3) that had been isolated from male urine. 

The earliest report of an isolated androgen was presented by Butenandt (10) in 1931. He isolated 15 mg of crystalline androsterone from 15,000 L of human male urine. A second crystalline compound, dehydroepiandrosterone, which has weak androgenic activity, was isolated by Butenandt and Dannenberg (11) in 1934. During the following year, testosterone was isolated from bull testes by David et al. (12). This hormone was shown to be 6 to 10 times as active as androsterone. 

The discovery of oestrone (VII) in 1929 by Doisy and independently by Butenandt was followed in 1934 by the isolation of progesterone (II) from corpus luteum tissue and of dehydroepiandrosterone (IV) from urine (see review by Petrow [16]). Although their chemical structures were not fully elucidated, a relationship between them and cholesterol was assumed. 

Fractionation of non-histone proteins 219 from rat leukaemia cells Isolation of dehydroepiandrosterone 220 sulphotransferase by affinity chromatography... 

Roberts et ah (1964) answered part of the question by injecting doubly labeled cholesterol sulfate into the splenic artery (supplying 90% of the blood to an adrenal carcinoma) of a female patient and isolating various steroid sulfates in the urine of the first 24 hours. Of the recovered radioactivity 0.46% was dehydroepiandrosterone sulfate, bearing the same H/ S ratio as the injected compound. Urinary androstenediol-3-monosulfate and 5-pregnene-3/3,17a,20a-triol-3-monosulfate as well as 16a-hydroxydehydroepiandrosterone-3-monosulfate also had the same ratio as the injected cholesterol sulfate, demonstrating that all these compounds have a common sulfated precursor, which could possibly be cholesterol sulfate. 

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