Ralph I. Dorfman

Etioallocholanol-3(β)-17-One (Isoandrosterone) as a Metabolite of Testosterone in the Human Male.∗

Continuing the study of the metabolic fate of testosterone in the human male, we have now been successful in demonstrating that etioallocholanol-3(β)-17-one (isoandrosterone) is a possible metabolite of testosterone. Previous communications have established androsterone and etiocholanol-3(α)-17-one as metabolic products of testosterone in the human male. 1 , 2 , 3 When testosterone was administered to the adult male guinea pig, etioallocholanol-3(β)-17-one was recovered in the urine. 4 The presence of etioallocholanol-3(β)-17-one has been established in the urine of a patient suffering from adrenal hyperplasia. 5 , 7 Recently, this substance has been isolated from the urine of normal women and also from the urines of both men and women suffering from cancer. 6 The urine was collected from a 30-year-old hypogonadal male during the time the individual was being injected intramuscularly with 30 mg of testosterone propionate† per day. During this period the daily excretion of androgenic substances exceeded 75 International Units (I.U.) per day compared to the value of 14 I.U. per day in the absence of any treatment. A total of 12 days of urine during this treatment was collected. The urine was extracted in the usual manner and the neutral compounds separated into the ketonic and nonketonic fractions by means of the Girard-Sandulesco reagent. 2 The neutral ketonic compounds were fractionated by means of chromatographic analysis using aluminum oxide (Brockmann) as the absorbent and carbon tetrachloride as the solvent. That fraction which was eluted with carbon tetrachloride containing 0.1% ethanol yielded a crystalline residue on evaporation of the solvent. This crystalline residue was dissolved in 4 cc of hot 90% ethanol, and 170 mg of digitonin in 8 cc of hot 90% ethanol was added.

The Fate of Testosterone in the Human.

Summary The conversion of testosterone to etiocholan-3(α)-ol-17-one in addition to androsterone in the human has been confirmed. It has been demonstrated that the conversion of testosterone to androsterone may take place in the complete absence of the testis.

Use of radiopotassium for detection of minute amounts of desoxycorticosterone

Conclusion Thus far only desoxycorticosterone has been studied with respect to potassium metabolism in the adrenalectomized animal. Under similar conditions it has been previously demonstrated1 that as little as one microgram produces a significant retention of sodium in the adrenalectomized rat. With radiopotassium 10 micrograms can be detected. This method affords a convenient way to evaluate the relative activity of various adrenal cortical steroids and adrenotrophic hormone. Further studies are needed to define the method as a quantitative assay for adrenal cortical steroids. Summary. By the use of radiopotassium in the adrenalectomized male rat, it is possible to detect as little as 10 μg of desoxycorticosterone. The method affords a convenient means of evaluating the relative activities of adrenal cortical steroids, as well as the influence of adrenotrophic hormone on potassium metabolism.

Comparative Activity of Naturally Occurring Estrogens on the Infantile Rat Uterus and Vagina.

In a previous report it was shown that there is a distinct difference between estrone and estriol in their respective abilities to cause hypertrophy of the infantile rat uterus and vaginal introitus. 1 These observations have been confirmed and extended by other workers to include estradiol. 2 , 3 In this communication we have further extended these comparative studies to include, in addition to estrone, α-estra-diol, and estriol, the benzoates of estrone and estradiol and also equilen and equilenin. The estrogens employed in this study were dissolved in olive oil so that the daily amount administered was always contained in 0.2 cc of oil. Twenty-five-day old female rats were injected subcutaneously once daily for 5 days. Twenty-four hours after the last injection, the animals were sacrificed. Body weight, uterine weight, and vaginal introitus were determined. At each concentration for each estrogen 4 or more animals were used, except in one instance (estriol at 55γ per day) when 3 animals were employed. The comparative uterine and vaginal activities of the estrogens studied are presented in Table I and Fig. I. α-Estradiol ranked as the most active estrogen in its ability to hypertrophy the uterus and produce vaginal introitus. As small a daily dose as 0.01 μg produced a 30% increase in uterine weight and 40% vaginal introitus. The descending order of uterine activity was α-estradiol, estrone, equilin, equilenin, and estriol. The descending order of activity on the vagina was α-estradiol, estrone, equilin, with estriol and equilenin being the least active of the group and of about the same magnitude. As previously described, the activity of estriol on the uterus is unique. Though a small dose stimulates the uterus, even a large dose (100γ) still does not hypertrophy the gland to its maximum size.

The antifertility activity of dl-cis-bisdehydrodoisynolic acid methyl ether☆

Abstract dl-cis-Bisdehydrodoisynolic acid methyl ether proved a highly effective antifertility agent when administered postcoitally in the rabbit. Like other estrogens, the compound lowered the carbonic anhydrase levels in the pregnant rabbit uterus. The lowered values appeared tobe dose related. Administration of 2 mg/day orally for 6 days following midcycle mating in the macaque monkey resulted in only one pregnancy in 98 positive matings. When given in a dose of 25 mg/day for 3 days starting on the 24–33 day of the cycle there were two pregnancies out of 41 matings. These represent significantly lower pregnancy rates than encountered in untreated matings in the primate colony.

Can Strophanthin Maintain Adrenalectomized Mice

In a recent report by Zwemer and Lowenstein it was suggested that adrenalectomized cats can be maintained in good condition by the administration of strophanthin. 1 This substance was administered daily in the concentration of 15 μg per kg of body weight. We have attempted to extend these findings to the mouse. For these experiments 21-day-old mice were bilaterally adrenalectomized. A total of 140 operated animals were employed, 40 serving as unin-jected controls, while the remaining 100 animals received various concentrations of strophanthin. The strophanthin was dissolved in olive oil so that the daily dose was contained in 0.1 cc of oil. Injections were made subcutaneously, beginning 24 hours after the operation and continued for 8 days. Table I summarizes the results of the administration of strophanthin in daily concentrations of 0.02, 0.2, 2.0, 5.0, 10.0, 20.0, 50.0, and 100.0 μg, respectively. Of 40 uninjected control animals, only 4 were alive on the 10th day after adrenalectomy. The average survival of the remaining 36 animals was 4.7 days. The administration of desoxycorticosterone acetate† in a concentration of 0.2 mg per day protected 15 of a group of 17 adrenalectomized mice. Strophanthin proved to be ineffective in the protection of the adrenalectomized mice in any of the 8 concentrations tested. No significant difference could be noted between the strophanthin injected and uninjected animals on either of the two criteria used; that is, number of animals alive 10 days post-operative, or the average survival of those animals dying before the tenth day. Strophanthin proved to be toxic in doses of 50 and 100 μg, respectively. All the animals were dead within 24 hours after the first injection of strophanthin at these concentrations.

Excretion of Sex Hormones in Urine of Adult Male Monkeys.4

The concentration of androgenic activity found in the urine of human males has been shown to be high in comparison to that found in the urine of stallions, bulls and rams. 1 It becomes of interest, therefore, to study the quantitative excretion of androgenic substances in the urine of the adult male monkey. Three normal adult rhesus monkeys of known fertility weighing between 6.9 and 10.0 kg were chosen for the urine collections. Four samples from each monkey were collected over periods of 72 or 96 hours. The method employed for the quantitative extraction of androgenic and estrogenic substances from the urines has been previously described. 2 The androgenic activity was assayed by means of the day-old chicks comb, 3 while the estrogenic activity was determined by means of the estrus reaction in the adult spayed female mouse. The results of the assays of both estrogenic and androgenic activity are listed in Table I. It is shown that the adult male monkey excretes both estrogenic and androgenic substances. The estrogenic activity varied from 11.1 to 2.5 I.U. per 24 hours, while the anidrogenic activity varied from 1.0 to 4.7 I.U. per 24 hours.

A NEW ESTROGENIC SUBSTANCE PRESENT IN THE NEUTRAL FRACTION OF HUMAN PREGNANCY URINE

In addition to the well-defined phenolic estrogens in human pregnancy urine there appears to be another estrogenic substance which is a neutral non-ketonic compound.