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Different natural and synthetic organic hydroperoxides have been found to stimulate TBARS formation in human term placental mitochondria. The levels of TBARS were lower than arising from NADPH-dependent lipid peroxidation. BHT, Mn2+ and DMPO counteracted TBARS formation in the presence of cumene hydroperoxide implicating involvement of free radicals in this process. On the other hand superoxide dismutase, catalase and EDTA while being inhibitory in NADPH-dependent lipid peroxidation did not inhibit cumene hydroperoxide-dependent TBARS formation. Amphenone B and SKF-525A, inhibitors of cytochrome P-450, strongly inhibit both NADPH- and cumene hydroperoxide-dependent lipid peroxidation. These data provide evidence that cytochrome P-450SCC is involved in both these processes. However NADPH-dependent lipid peroxidation and the cumene hydroperoxide have been found to inactivate placental mitochondrial cytochrome P-450SCC. The presence of cumene hydroperoxide resulted in a more rapid inactivation of cytochrome P-450SCC and consequently inhibited NADPH-dependent lipid peroxidation. It has been observed for the first time that progesterone biosynthesis can be inhibited by cumene hydroperoxide. Protective effect of Mn2+ and DMPO on progesterone biosynthesis indicates the importance of free radicals as transient products of cytochrome P-450SCC-dependent cumene hydroperoxide metabolism. In contrast to progesterone formation from cholesterol, the conversion of pregnenolone to progesterone was not affected by cumene hydroperoxide. This suggests that inhibition of progesterone synthesis from cholesterol by hydroperoxide may be ascribed to its effect on the desmolase activity of cytochrome P-450SCC in placental mitochondria. On the basis of the results obtained, we propose that the inhibition of progesterone biosynthesis by naturally occurring hydroperoxides may contribute to the development of preeclampsia.

When paraquat was incubated with mouse brain microsomes in the presence of NADPH, a Nash-reagent-reactive substance (NRRS) (but not formalin) was produced. It was found that NRRS production was decreased in a dose-dependent manner by N-ethylmaleimide, a sulfhydryl reagent, which also inhibited NADPH-cytochrome P-450 reductase in parallel with the decrease in NRRS production. NRRS production was reduced by radical scavengers (catechin, glutathione, mannitol, Superoxide dismutase and catalase). or under anaerobic conditions. In addition, inhibitors of adrenal cortex mitochondrial cytochrome P-450 (metyrapone, aminoglutethimide and amphenone B) inhibited NRRS production without causing a significant decrease in NADPH-cytochrome P-450 reductase activity. These findings suggest that active oxygen species and the mixed-function oxidase system may play important roles in NRRS production from paraquat in brain microsomes.

The NADPH-dependent lipid peroxidation in human placental mitochondria has been found to be inhibited strongly by amphenone B, aminoglutethimide and carbon monoxide, inhibitors of cytochrome P-450-mediated reactions, but was hardly affected by respiratory chain inhibitors. Cytochrome c, an exogenous electron acceptor which is known to compete with cytochrome P-450 for the reducing equivalents, showed an inhibitory effect on NADPH-dependent lipid peroxidation. The observed NADPH-dependent superoxide generation was also strongly inhibited by amphenone B and aminoglutethimide. Moreover, the lipid peroxidation in placental mitochondria was demonstrated to be stimulated by xanthine/xanthine oxidase added as superoxide generating system. This peroxidation was not affected by amphenone B and aminoglutethimide. On the other hand, the superoxide dismutase was found to inhibit both the xanthine oxidase- and NADPH-dependent lipid peroxidation. These data provide evidence that cytochrome P-450 is involved in NADPH-dependent mitochondrial lipid peroxidation. It is suggested that superoxide liberated from cytochrome P-450, in combination with iron, may be responsible for initiation of NADPH-dependent lipid peroxidation in human placental mitochondria.

1. By spectrophotometric determination the content of cytochrome P450 in the microsomal preparation from Tetrahymena pyriformis (strain WH-14) was estimated to be less than 0.014 nmol/mg protein. Optical absorption spectroscopy at 77°K revealed that the microsomal cytochromes mainly consist of b-type heme protein. 2. The microsomal NADPH-oxidase activity was inhibited by hydroxylase inhibitors such as metyrapone, amphenone B and elipten phosphate to a much lesser extent compared with rat liver microsomal activity. 3. No significant hydroxylation activity of xenobiotics by the protozoan microsomal sample was detected. The activity was not induced by administration of phenobarbital or 3-methylcholanthrene. 4. Nonparasitic protozoon Tetrahymena pyriformis appears to have very low if any cytochrome P450-dependent detoxication mechanism. These observations are in contrast to the observation by Agosin et al., 1976, who reports a weak but detectable activity of drug hydroxylation in parasitic protozoon Trypanosoma cruzi.

Metyrapone and SKF-525A, together with amphenone B, a structural analogue of metyrapone, which are all inhibitors of cytochrome P-450-mediated reactiors, were shown to inhibit the arachidonic acid-induced aggregation of human platelets. Amphenone B, like metyrapone, exhibited a type II (ligand) binding spectrum with rat liver microsomal cytochrome P-450, in contrast to SKF 525A which is a type I (substrate) binding agent. Independently of their type of binding spectra and of their maximum spectral change, however, the affinity of the three compounds for rat liver cytochrome P-450 showed a close proportional correlation with their platelet aggregation inhibitory potency. All three compounds inhibited the formation of [1−14C]thromboxane B2 from [1−14C]arachidonic acid by human platelets aggregated with collagen. The effect of metyrapone on the remaining labelled products suggested that it is a selective thromboxane synthesis inhibitor, while amphenone B exhibited activity reminiscent of cyclo-oxygenase inhibitors. SKF 525A produced complex effects possibly attributable to cyclo-oxygenase inhibition and enhanced lipid peroxidation, since it also enhanced platelet malonaldehyde formation, which the other two compounds inhibited. These data provide further support for a role of cytochrome P-450 in thromboxane synthesis and platelet aggregation.

(1) In order to study the relationship between aromatization (estrogen biosynthesis) and 1beta-hydroxylation, the effects of a variety of factors on these processes were evaluated. (2) Using the C18 substrate, 4-estrene-3,17-dione, it was found that carbon monoxide, SU-4885, amphenone B, potassium cyanide, 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione inhibited the above transformations significantly and to varying degrees. However, within a given experiment the inhibition of each process was similar. (3) SKF-525A did not inhibit either transformation. In addition, phosphate, Tris and barbital buffers, as well as pH changes from 6.9 to 7.7, had no stimulatory or inhibitory effect on the production of estrogen and 1beta-hydroxy compounds. (4) In contrast, several inhibitors affected the aromatization of C19 and C18 steroids differently. These include carbon monoxide, SU-4885 and amphenone B. (5) When a mixture of 4-[7beta-3Hi1estrene-3,17-dione and 19-[4-14C]nortestosterone were incubated together the former was preferentially converted to estrogen. This preference for the 17-keto steroidal form mimics results observed for C19 substrates. (6) We conclude that while estrogen biosynthesis and 1beta-hydroxylation appear to be mediated by the same enzyme system, the same conclusion cannot be drawn for the aromatization of C19 and C18 substrates.

The effect of Amphenone B, an inhibitor of corticoid synthesis, on thyroxine (T4)-induced metamorphosis was studied in toad tadpoles kept in thiourea. Amphenone injections retarded T4-induced tail resorption markedly. The effect of Amphenone was nullified by aldosterone and corticosterone added to the water in which tadpoles were kept. Steroidogenic cells of adrenals in Amphenone-injected animals were enlarged markedly as compared with those in the saline-injected tadpoles or the Amphenone-injected tadpoles which were supplemented with corticoids. The results strongly suggest that endogenous corticoids act together with thyroid hormone to accelerate metamorphosis.

In short termexperimentsinthedog2-methyl–l,2-bis(3-pyridyl)-l-propanonc (SU 4885) in high dosage totally suppressed the secretion of steroids by the adrenal gland. In low dosage there was a shift from cortisol and corticosterone to 11-desoxycortisol (Reichstein's Compound S) and desoxycorticosterone suggesting a selective inhibition of hydroxylation at C11. In contrast amphenone suppressed total steroid secretion (i.e., of cortisol and corticosterone) with no evidence for a selective action on 11-hydroxylation.

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The effect of Amphenone B on development of renal hypertension was studied in rats whose kidneys were encapsulated with latex envelopes. The compound was administered in the diet for 13 weeks at dose levels of 0.1 and 0.3% by weight. Both dose levels of Amphenone B were equally effective in preventing rise of systolic blood pressure to the level of untreated, renal-encapsulated controls although both doses of the drug allowed modest rises of pressure to occur. Growth was inhibited by the drug. Amphenone B depressed thyroid activity as assessed by increased release rate of radioactive iodide from the thyroid gland; decreased resting metabolic rate; decreased growth; decreased food intake; and increased thyroid size. The protection against development of hypertension afforded by this compound may be related, at least in part, to its antithyroid activity.

SummaryWhen compared with appropriate controls, it appears as if rats treated with Amphenone “B” show a decrease in function of the hypertrophied thyroid gland as measured by oxygen consumption but a probable increase in cortisone-like function of the hypertrophied adrenal gland as measured by electroshock seizure threshold, thymus involution, reaction to 24-hour fast? and histological study of the adrenal.

The preparation of 13 p-substituted α,α-dimethyldesoxybenzoins is given. In view of their adrenocortical activities the diamino compound II (isomer of Amphenone B) and the two monoamino derivatives IV and XI are of special interest.

Amphenone B was added in concentrations of 0.1 to 0.5% to a number of synthetic diets and to a fox chow diet. These were then fed to young male rats to determine the effects on adrenal weight and on the cholesterol content of the adrenals, liver, and plasma. The rats fed amphenone mixed with synthetic diets showed a greater increase in adrenal size and adrenal cholesterol than those fed amphenone mixed with the chow diet. The liver and plasma cholesterol values of rats fed amphenone on synthetic diets were also increased appreciably while those of rats fed amphenone in the chow diet were near normal. Synthetic diets containing erucic acid and amphenone appeared to have the greatest effect on adrenal size and cholesterol content but other synthetic diets in some cases caused a greater increase in liver and plasma cholesterol concentrations. An attempt to examine the effect of injected amphenone on cholesterol levels in rats fed different diets was limited by the fact that high doses of amphenone cause prolonged anesthesia.

SummarySuppression of corticosteroid production by inhibitors effected a tenfold increase in susceptibility of mice to death from endotoxin shock. Amphenone B and metyrapone caused some increase in susceptibility, but if given more than 1 hr before the endotoxin afforded protection to the test animals. Aminoglutethimide exerted a deleterious effect for at least 24 hr. The effect of the aminoglutethimide was reversed by pregnenolone, progesterone and deoxycorticosterone, although the LD50 remained lower than in the control animals. Corticosterone, cortisone and hydrocortisone afforded marked protection in both the endotoxin treated mice as well as the endotoxin-inhibitor treated mice. Andros-tenedione caused a decrease in the LD50 value in both the control and aminoglutethimidetreated mice. By contrast 17-a-hydroxyprogesterone afforded no protection against endotoxin in the control mice and a significant, but miniscule, degree of protection in the inhibitor-treated mice.This investigation was supported by ...

The possibility is discussed of inhibiting the adrenocortical functions either selectively or preferentially by blocking the hydroxylations in positions 11β, 17α, 18 and 19 of the steroid nucleus or in 20α, 21 and 22 of the side chain, which are necessary for the formation of the individual corticosteroids from cholesterol. Many compounds with or without other biological activities, such as o,p′-DDD, Amphenone B, various blockers of cholesterol biosynthesis, heparinoids and various heterocyclic compounds, are shown to be devoid of a specific blocking activity on the adrenal cortex in vitro. However, amongst a large group of glutaric imides a few compounds proved to be active, mainly by blocking the 20α-hydroxylation of cholesterol, the most active being a cyclic hydrazide; the secretion in vivo of corticosteroids greatly decreased in response to 3–20 mg/kg. A new class of compounds active in vitro was found among simple imidazole derivatives, which block 17α, 18 and 19-hydroxylation.

Die Hoden geschlechtsreifer Rattenbocke wurden ohne Vorbehandlung nach Hypophysektomie, nach Verabfolgung von Choriongonadotropin (Primogonyl) und von Amphenon B elektronenmikroskopisch untersucht. Das Cytoplasma normaler Zwischenzellen besitzt ein blasiges, glattes ER, zahlreiche Mitochondrien vom gemischten Typ mit elektronendichten Einschlussen, freie Ribosomen, einzelne Fetttropfen und eine typische, gut ausgebildete Golgizone. Das blaschenformige ER wird offenbar von den Golgihohlraumen abgeschnurt. Nach Hypophysektomie sind kaum Veranderungen wahrzunehmen. Nach Choriongonadotropin nimmt die Zahl und Grose der Blaschen zu, die Mitochondrien einzelner Zellen schwellen und die Mitochondrieneinschlusse verschwinden. Nach Amphenon B-Gabe geht das vesikulare ER in ein tubulares uber. Die Golgihohlraume zeigen keine Erweiterungen. Diese Veranderungen werden mit den biochemisch fasbaren Hemmungen oder Stimulierung einzelner Syntheseschritte des Testosterons in Zusammenhang gebracht.

A testicular microsomal fraction containing the enzyme systems necessary for the production of testosterone from pregnenolone was divided in two subfractions by sucrose density gradient centrifugation in the presence of CsCl. When the subfractions were examined by electron microscope, one consisted mainly of smooth-surfaced particles, the other of rough-surfaced particles with ribosomes on their outer surface. Enzymic and spectrometric investigation of these subfractions revealed that most of the enzyme activities related to the androgen synthesis, which remained after the gradient centrifugation, and also of the cytochrome P-450, which would be a direct site of activation of molecular oxygen for the 17α-hydroxylation and the side chain cleavage, were localized in the smooth-surfaced submicrosomal fraction. The enzyme activities of the submicrosomal fraction which were diminished by the gradient centrifugation were largely restored by addition of the heated 105,000 × g supernatant fluid. Enzyme inhibitors such as Amphenone B (3,3-bis ( p -aminophenyl)-2-butanone dihydrochloride) and SKF-525A (2-diethylaminoethyl 2.2-diphenylvalerate hydrochloride) inhibited the testicular microsomal 17α-hydroxylase and the C 17 –C 20 lyase competitively. The apparent inhibition constants of Amphenone B were 9.17 × 10 −5 M against 17 α-hydroxylase and 2.45 × 10 −4 M against C 17 -C 20 lyase, and those of SKF-525 A were 1.61 × 10 14 M against 17α-hydroxylase and 1.28 × 10 −5 M against the C 17 –C 20 lyase.

AMPHENONE “B”3 (Allen and Corwin, 1951), a substituted desoxy- benzoin, given orally or subcutaneously, produces marked enlargement of rat thyroid and adrenal glands (Hertz, Allen and Tullner, 1950; Hertz, Tullner and Allen, 1951; Hogness, Williams and Lance, 1952). The thyroid enlargement is associated with reduction in I131 concentration (Hogness, Williams and Lance, 1952) and a lowered metabolic rate (Holtkamp, Heming and Sax, 1952). The thyromegalic effects are not found in hypophysectomized rats (Hertz, Allen and Tullner, 1950) or in those given supplementary thyroid extract (Hertz, Tullner and Allen, 1951). The adrenomegaly produced by Amphenone “B”is associated with increase in cholesterol concentration (Hogness et al., 1952) without evidence of decreased adrenal function (Hertz et al., 1951; Hogness et al., 1952; Heming et al., 1952). As in the case of the thyroid, adrenal enlargement is not found in hypophysectomized animals (Hertz et al., 1950) or in those given cortisone (Hertz et al., 1951).

We have investigated the effect of paraquat (methyl viologen) on lipid peroxidation in bovine adrenal cortex mitochondria. Incubation of a buffered aerobic mixture of mitochondria in the presence of Fe2+ or NADPH resulted in the formation of lipid peroxides whose accumulation could be followed at 532 nm as malondialdehyde. Fe2+ stimulates lipid peroxidation in normal mitochondria and those in which enzymes have been inactivated with heat. In contrast, NADPH has a stimulatory effect only in normal mitochondria, but not in heat-treated mitochondria. These results indicate that NADPH-dependent lipid peroxidation is an enzymatic process. Paraquat strongly inhibits this enzymatic lipid peroxidation, but has no effect on the non-enzymatic Fe2+-dependent process. The chemiluminescence that accompanies the NADPH-dependent lipid peroxidation is also markedly decreased in the presence of paraquat. Superoxide dismutase, which removes Superoxide anion efficiently, does not inhibit malondialdehyde production. The mechanism of the inhibition of the lipid peroxidation by paraquat has been examined. Paraquat has no effect on NADPH-2,6-dichlorophenolindophenol reductase and on NADPH-cytochrome c reductase activities in bovine adrenal cortex mitochondria. However, paraquat strongly inhibits the NADPH-dependent reduction of cytochrome P-450. These results suggest that the inhibitory effect of paraquat on NADPH-dependent lipid peroxidation in adrenal cortex mitochondria is due to a decrease in the level of reduced cytochrome P-450 probably by diverting electrons from cytochrome P-450. Cytochrome c, which can compete with P-450 for available electrons from adrenodoxin, like paraquat had an inhibitory effect on NADPH-dependent lipid peroxidation. Lipid peroxidation was also strongly inhibited by steroid hydroxylase inhibitors e.g., amphenone B, aminoglutethimide and metyrapone.

Using sonicated mitochondria fraction prepared from bovine adrenal glomerulosa cells, aldosterone biosynthesis from 18-hydroxycorticosterone was examined as its final step, as production of [ 3 H]-aldosterone from [ 3 H]-corticosterone was strongly reduced by addition of non-radioactive 18-hydroxycorticosterone during the incubation. Significant conversion of 18-hydroxycorticosterone to aldosterone by the mitochondria sonicate was observed in the presence of NADPH, but not NADP + . This reaction was almost completely inhibited in the atmosphere of 100% carbon monoxide in the presence of either NADP + , or NAD + , and significantly reduced in the mixture of carbon monoxide and oxygen (90:10) in the presence of NADPH. Several drugs, such as SU compounds, spironolactone, amphenone B and SKF 525A which affect cytochrome P-450 blocked production of aldosterone from 18-hydroxycorticosterone. From these results, we conclude that a mixed function oxidase involving a cytochrome P-450 is engaged in the final course of aldosterone biosynthesis.

The in vitro and in vivo methods are described that are used in the investigation of the effects of drugs on the biosynthetic pathways of steroid hormones from cholesterol. The sub-cellular locations of the enzymes involved in steroidogenesis are also. summarized and the effects on their activity of the following compounds are explained: o,p'-DDD, Amphenone B, Metyrapone. SU-8000, SU-9055, SU-10'603, Doriden ® , o-aminoglutethimide, Elipten ® , SKF-525A, SK & F-12185, Ba-40'028 and cyanotrimethylandrostenolone. Some attempts are made to correlate their histological effects on mitochondria and endoplasmic reticulum with their inhibitory action on enzymes that are associated with these sub-cellular sites. The evidence for the binding of Metyrapone to cytochrome P-450, involved in the action of the “mixed function oxidases”, is reviewed. Finally, the uses of the various inhibitors of steroid hormone biosynthesis in patients suffering from endocrine diseases are assessed.

Incubation of 18-hydroxycorticosterone with the sonicated mitochondrial preparation of bovine adrenal glomerulosa tissue leads to the production of aldosterone, as measured by radioimmunoassay. The in vitro production of aldosterone from 18-hydroxycorticosterone requires both molecular oxygen and NADPH, and is inhibited by carbon monoxide. Cytochrome P-450 inhibitors such as metyrapone, SU 8000. SU 10603, SKF 525A, amphenone B and spironolactone decrease the biosynthesis of aldosterone from 18-hydroxycorticosterone. These results support the conclusion that the final reaction in aldosterone synthesis from 18-hydroxycorticosterone is catalyzed by an oxygenase, but not by 18-hydroxysteroid dehydrogenase. By the same preparation, the production of [3H]aldosterone but not [3H]18-hydroxycorticosterone from [1,2-3H ]corticosterone is decreased in a dose-dependent manner by addition of non-radioactive 18-hydroxycorticosterone.

In concert with studies of the effects of various pharmacologic inhibitors of corticosteroidogenesis on adrenocortical morphology, U-8113, an analog of amphenone B, was administered daily to Sprague-Dawley rats for 7, 14, 21 or 30 day. The primary morphological responses involved increases in adrenal weight, width of zona fasciculata, width of zona reticularis, intracellular lipids, mitochondrial size, mitochondrial vacuolation and crystalline-like inclusions, small coated vesicles, lysosomes, autophagic vacuoles and cholesterol ester clefts. In particular, the increases in lysosomes, coated vesicles and autophagic vacuoles containing morphologically altered mitochondria were considered reflective of mechanisms designed to maintain cellular integrity amidst functional impairment. Lipid analysis revealed marked increases in cholesterol esters and phospholipids, supportive of morphological observations. When permitted a 14 day recovery period following either 14 or 30 days of inhibitor therapy, most fine structural alterations and lipid derangements were diminished, and the cells approximated normal parameters.

PREVIOUS publications have described the chemical synthesis (Allen and Corwin, 1950) and some of the biological properties of Amphenone-B.* This substance has progestational activity in the rabbit (Allen, Hertz and Tullner, 1950) and has an anesthetic effect in the rat, rabbit and dog (Hertz, Allen, Tullner and Westfall, 1952). Amphenone-B also has folliculoid activity (Hertz, Allen and Tullner, 1950) in the castrated female rat. It has been reported, in addition, that the rat responds to the compound with a marked hyperplasia of the thyroid, a goitrogenic action which is probably related to its aromatic amino groups (Hertz et al., 1951). But of all the effects in its bizarre spectrum of endocrine action, the most interesting and unexpected is the profound hypertrophy of the adrenal glands. Just as the goitrogenic activity can be suppressed by the simultaneous administration of thyroxine, the hypertrophy of the adrenals can be suppressed by the administration of cortisone (Hertz, Tullner, and Allen, 1951).

Western painted turtles ( Chrysemis picta belli ) were adrenalectomized to show if adrenocortical steroids are necessary for electrolyte homeostasis. Two weeks after removal of the adrenal glands plasma Na had decreased by 13.7% whereas plasma K had increased by 22.1%. There were no significant changes in plasma Ca, Mg, or Cl. Plasma osmolarity increased by 4.4% while the blood hematocrit decreased by 31.9%. Muscle Na and Cl decreased by 27.1% and 26.7%, respectively, but there was no change in muscle K or in percentage of muscle water. Electrolyte changes observed 3 weeks after hypophysectomy were similar to those observed 2 weeks after adrenalectomy Treatment with Amphenone B or betamethasone was followed by some electrolyte changes, but injection of Metopirone failed to impair electrolyte balance. The only measurable response with aldactone was a significant increase in plasma calcium. Hepatic glycogen concentrations decreased after hypophysectomy and adrenalectomy and increased after treatment of intact turtles with betamethasone; Amphenone B had no effect.

The polarographic behaviour of the compounds Amphenone B (3,3-bis(p-aminophenyl)-butan-2-one) and Su 4885 (1,1-dimethyl-2-oxo-ethylenebis-3-pyridine) in aqueous medium is described. Both substances are reduced at the dropping mercury electrode by two electrons and produce well-developed diffusion cathodic waves of a height proportional to the concentration of the depolariser. Su 4885 can be estimated polarographically also in adrenal homogenates in Ringer-Krebs phosphate buffer.

SummaryAmphenone “B” induces a marked hypertrophy of both the thyroid and adrenal in the rat. Simultaneous administration of thyroxin and Amphenone “B” prevents the thyroid hypertrophy without reducing the adrenal enlargement. Combined cortisone and Amphenone “B” administration prevents the adrenal enlargement without altering the thyroid hypertrophy. Thus, Amphenone “B” exerts its adrenal and thyroid effect by acting through two distinct end—points in the anterior pituitary.

Summary(1) Enlargement of rat adrenal and thyroid glands following the administration of Amphenone “B”, observed by Hertz, Allen, and Tullner, was confirmed. (2) The adrenal enlargement was associated with an increase in adrenal cholesterol concentration. At the same time, the adrenals remained capable of secreting sufficient steroid to produce a marked fall in circulating eosinophiles. Amphenone “B” may exert its adrenomegalic effect by direct stimulation of the adenohypo-physis to produce ACTH rather than indirectly as seen, for example, in thiouracil-induced goiter. (3) In addition to thyroid enlargement, a marked decrease in I131 concentration in the gland was observed. This could be due to a thiouracil-type of action, although the effects noted within the time relationships of our experiments were similar to those produced by TSH and hence possibly the result of increased secretion of radioiodine rather than a decreased uptake. (4) Decrease in size of the testes was observed in animals given Amphenon...

A simple aerobic and anaerobic (N 2 plus TTC) perfusion technique was employed to study the intermediary metabolism of intact calf adrenals and the effect of disparate chemical and physical agents on their metabolic activity and steroidogenic function. The results demonstrated that Meyerhof-Embden and Krebs-Johnson carbohydrate intermediates were rapidly oxidized in contrast to amino and fatty acids. Studies with various selective enzyme inhibitors confirmed the role of the glycolytic and terminal aerobic pathways. The nature and intensity of the effects of these agents on the steroidogenic processes paralleled those on the metabolic activity with the striking exception of Amphenone B, which produced only a minor decrease in the oxidative metabolism and one of 75% in corticoid output. A 24-hr. incubation at 8 °C. and exposure to γ-radiation (1900–2700 r.) revealed a similar dissociation between their effects on the metabolic and steroidogenic activities of the adrenal. Thus, while a pronounced decline in metabolic activity was invariably associated with a diminution in biosynthetic function, the latter can be markedly depressed without a corresponding depression in the former.

SummaryIntravenously administered Amphenone “B” produces surgical anesthesia in the rabbit and dog. The margin of safety is sufficient to permit complete anesthesia without major respiratory depression and with rapid recovery. The combination of endocrinological and anesthetic properties of Amphenone “B” parallels that previously observed for progesterone and other steroids.

The precise pathogenesis of ascites in cirrhosis of the liver remains obscure. Portal hypertension, 1 , 2 decreased plasma osmotic pressure due to hypoalbuminemia, 3 , 4 increased lymphatic exudation from the liver, 5 , 6 increase of antidiuretic hormone, 7 and increase of sodium-retaining adrenocortical hormone 8 are among the factors considered in the pathogenesis of ascites. The marked decrease of sodium excretion and the presence of increased amounts of aldosterone in the urine of patients with cirrhosis and ascites 9 point to this sodiumretaining hormone as the possible culprit in the pathogenesis of ascites. The complexity of the pathogenesis of ascites is demonstrated schematically in Figure 1. Amphenone B (3,3-bis[ p -dimethylaminophenyl]butanone-2-dihydrochloride) (Fig. 2) was synthesized by Allen and Curwin, 11 in 1950, and has been shown to have principally an inhibiting effect on thyroid and adreno-cortical function. 12,13 Among its other effects are a progesterone- or estrogen-like effect on the female genitalia, with uterine enlargement 12 and a depressing effect on the central