16α-Hydroxysteroids. X.¹ 2β-Hydroxylation of 9α-Fluorohydrocortisone by Streptomyces roseochromogenus

Abstract

2859described18 using a Neurospora sp. (M 714). The methylene chloride extracts were concentrated to dryness, dissolved in 10% ethyl acetate in benzene and chromatographed on 1400 g. of silica gel. The chromatographic column was eluted successively with 10%, 12%, 15%, 25%, 35%, 40%, and 50% ethyl acetate in benzene. The 50% eluates were concentrated to dryness and the residue was crystallized from acetone,cyclohexane, then from methanol, and finally from acetonecyclohexane again to yield 394 mg. of 7a-hy droxy-4-androstene-3,17-dione, m.p. 255-256.5*, C°°2 41 m", « 16,000.The maximum in the ultraviolet spectrum in 0.101Y methanolic potassium hydroxide shifted on standing from 241 mg to 283 µ.

Hormones

“…Selective hydroxylation of the double bond at position 16 with osmium tetroxide in pyridinebenzene gives compound 127. The 9,11-double bond of this compound is converted to a 9α-fluoro-11β-hydroxyl group (128) by the method described for 9α-fluorocortisol (123) The 16α-hydroxyl group can also be introduced directly into 9α-fluoroprednisolone (124) by using a microbiological method originally developed by Squibb [253,254]. Triamcinolone 16α,17α-acetonide (130) is prepared by stirring a suspension of triamcinolone (129) in acetone in the presence of catalytic amounts of perchloric acid at approximately 20…”

16α-Hydroxysteroids. XIII.* Carbonyl Reduction by Streptomyces roseochromogenus

“…the aldehyde IV does not appear to be a biological intermediate in the formation of 21-acetylamino-9 -fluoro -11,3,17 -dihydroxy -4 -pregnene-3,20-dione from I by S. roseochromogenus (Smith et al, 1962). The aldehyde IV is not hydroxylated by S. roseochromogenus; neither 16 a-nor 2/3hydroxylated derivatives could be detected.3 Experimental 9a-Fluoro-l 1ß,17 a,203,21 -tetrahydroxy-4-pregnen-3-one (II).-The steroid-rich broth extract concentrate obtained from S. roseochromogenus fermentations of 9a-fluorohydrocortisone already described (Smith et al, 1961) was used for the isolation of 20/3-dihydro-9a-fluorohydrocortisone. An initial separation of some non-steroidal materials was accomplished by adsorption of the extract concentrate onto silica gel from ethyl acetate.…”

“…In our studies of the 16 -hydroxy latió n of 9afluorohydrocortisone (I) by Streptomyces roseochromogenus a variety of reducing (Smith et al, 1960b;Smith et al, 1961) and non-reducing (Smith et al, 1962) steroids have been isolated and identified.…”

“…Isolation of II from a selected steroid-rich broth extract concentrate previously described (Smith et al, 1961) was accomplished by paper chromatography, and the steroid thus obtained was identified as 20/3-dihydro-9a-fluorohydrocortisone1 by comparison of infrared spectra, chromatographic behavior, and color tests. Attempts at isolation of the very polar III did not give a satisfactory preparation for characterization.…”

16α-Hydroxy Steroids

“…We have found that Streptomyces roseochromogenes, in addition to hydroxylating 9a-fluorohydrocortisone in the 16a-position (Thoma et al, 1957), is able to affect hydroxylation in the 2,3-position to yield both 2(3-hydroxy-9a-fluorohydrocortisone and the 2#3, 16adihydroxy-9a-fluorohydrocortisone. The isolation and structure proof of the 2,B-hydroxylated products are presented elsewhere (Smith et al, 1961). The degree of 2,B-hydroxylation was found to be highly strain dependent.…”