The Bio-Oxygenation of 11-Desoxycorticosterone at C-11

Abstract

Communications to the Editor 3261 Preliminary fractionation of the benzene-extractable alkaloids by the excellent procedure of Jacobs and Craig4 yielded the five known crystalline alkaloids and a large amorphous fraction; only the latter was active as a hypotensive agent at low dosage. Further fractionation of this material guided by assay3 in hypertensive patients yielded a highly active concentrate, which on 24-plate Craig distribution, using 2 M acetate buffer at pH 5.5 and benzene as the immiscible phases, exhibited two discrete peaks. The material having a peak at tube 15 (K = 1.67) showed activity when administered orally in doses of 0.6-0.8 mg. per patient, while the material from the second peak, at tube 6 (K = 0.35), was active at about 4 mg.These fractions crystallized readily from methanol-water and ethanol-water respectively, yielding germidine (m. p. 220-223°( cor.); [a]25D + 13°( c, 1.67 in chloroform)) and germitrine (m. p. 197-199°( cor.); [ct]25D + 11°( c, 1.54 in chloroform)).Room temperature hydrolysis of germidine with 0.1 N aqueous methanolic alkali afforded germine (C27H43O8N),6 acetic acid and a-methylbutyric acid. The former was identified by rotation, analysis and conversion into monoacetonylgermine hydrochloride.6 The acids, after conversion into the p-phenylphenacyl esters followed by chromatography on alumina, yielded p-phenylphenacyl -methylbutyrate7 (m. p. 71-72°( cor.)) and pphenylphenacyl acetate8 9(m. p. 109-110°( cor.)). Analysis of the free base and the crystalline thiocyanate (m. p.242-2440 dec. (cor.)) indicates germidine to be an ester of germine with one mole each of the above acids.Hydrolysis of germitrine yielded germine, amethylbutyric acid and methyl-ethylglycolic acid. The phenylphenacyl ester of the latter (m. p. 119-120°( cor.); [q]26d + 5°( c, 0.64 in chloroform))was identical with an authentic sample.Analysis of the free base and the thiocyanate (m. p. 228-231°dec. (cor.)) indicates that germitrine is probably a mono--methylbutyrate dimethylethylglycolate of germine.The specific rotations of the branched-chain acids (isolated from the total amorphous fraction because of an insufficient supply of the crystalline alkaloids) showed these materials to be l-a-methylbutyric and d-methylethylglycolic acids. Germidine and germitrine, injected intravenously in doses of 0.6-0.8 y per kg., markedly lowered the blood pressure of anesthetized dogs and cats.

New assumptions about oxidative processes involved in steroid hormone biosynthesis: Is the role of cytochrome P-450-activated dioxygen limited to hydroxylation reactions or are dioxygen insertion reactions also possible?

“…Cytochrome P-450's can catalyze the insertion of oxygen between a C atom and a hydrogen atom [1]. Insertion of • OOH into a C atom bearing a tertiary H atom (e.g.…”

The Chemistry and Metabolism of the Steroid Hormones

“….perfusion of the isolated beef gland with 11-desoxycorticos terone led to the production of corticosterone (113). This ability of the gland to effect l1-oxygenation is not confined to ll-desoxycorticosterone since 11-hydroxyandrosterone has been isolated after perfusion with androsterone and 17 -hydroxycorticosterone after perfusion with 17a-hydroxy-ll-desoxy corticosterone; l1-oxygenation of other steroids (e.g., progesterone, 17a hydroxyprogesterone) is indicated (114). Since this l1-oxygenation is effected in the absence of ACTH, and is not materially enhanced by the addition of ACTH to the perfusate, this step in the biosynthesis of typical adrenocor ticosteroid is obviously beyond ACTH control.…”

“…Employing a perfusion apparatus recently described (111), Hechter has ob served the increased production of formaldehydogenic neutral lipid and con comitant glycogenic activity after the addition of ACTH to the perfusion medium (112) . .perfusion of the isolated beef gland with 11-desoxycorticos terone led to the production of corticosterone (113). This ability of the gland to effect l1-oxygenation is not confined to ll-desoxycorticosterone since 11-hydroxyandrosterone has been isolated after perfusion with androsterone and 17 -hydroxycorticosterone after perfusion with 17a-hydroxy-ll-desoxy corticosterone; l1-oxygenation of other steroids (e.g., progesterone, 17a hydroxyprogesterone) is indicated (114).…”

“…Dobriner and co-workers (89), have considered l1-oxygenated steroids, such as ll-hydroxyandrosterone, as specific adrenocorticosteroid catabolites, and it is noteworthy that this particular compound appears to be excreted largely unchanged after administration to an Addisonian patient (127). Nonetheless, the finding of Hechter et al (114) that the adrenal may effect l1-oxygenation of androsterone and other nonadrenal steroids reopens the question of the probable origin of l1-oxygenated compounds.…”

The Conversion of Pregnenolone to a More Active Progestational Substance by Incubation With Endocrine Tissues in Vitro

“…The successful experiments reported by a number of workers [Hechter, Jacobsen, Jeanloz, Levy, Marshall, Pincus & Schenker, 1949, 1950Hayano, Dorfman & Prins, 1949;Nissim, 1952], demonstrating the ability of the adrenals to convert mineralocorticoids into gluco-corticoids, suggested that similar enzymic transformations of steroids might take place in other endocrine organs. Therefore, an investigation of the capacity of the corpus luteum to synthesize progesterone from other steroids was undertaken.…”