productc is provided by the action of lithium aluminurn hydride on this intermediate \r.7 Acknowledgment.--The authors are grateful to Dr. H. T. Openshaw for a sample of isoemetine hydrobromide. Financial support was provided by the research committee of the Graduate School.
genus." The flasks were incubated at 25' for 4 to 7 days with rotatory mechanical shaking a t ?SO r.p.in. in a 2 in. radius, the niyceliutn filtered off arid washed with water. The filtrate was extracted with methyl isobutyl ketone (three 800-ml. portions), the extract concentrated to small volume, cooled and the resulting crystals recrystallized ( 7 ) I t is pertinent t h a t in t h e case of t h e A3-dehydroyohimbane (D-E f r o n s ) a n d similar ( c j . ref. 4) systems, catalytic reduction also from ethyl alcohol. The properties of IT1 (m.1'. afford? t h e more stable 3,15,20-cis-fvans product (E. E . v a n Tarnelen, %0-252', [ a ] +97' ~ (c 0.99 in pyridine); x$iSX 11.S h a m m a and P. E. Aldrich, TIIISJCXJRXAI., 78, 4628 (195G). 238 rnp (15,000); '?;:A 2.79, 2.98, 5.82, 6.00, I h P A R i X I E N T O F CHEMISTRY EI'GESE E.Y A X TABIELES6 . 1 7 ~ ~ Anal. ; Found: C, G3.7i; H, '7.32) and 117-SIVERSITY O F ~ v I S C O N S I N PAUL E. ALDRICH (m.p. 248-250'; [ c u ] ? ~ D+ 7 l o (c 0.35 in acetone); ir.\nIs(lh-, W I S C O S S ~ ~ Jxcxsos R . HEWER,J R . A $ " . : 2.95, 5.85, 6.02, 6.16, 6.24, 11.26 p ; RECEIVED J r S E 17, 1%'T diacetate: n1.p. 170-180" (with gas evolution) ; [ a I z 3 +28" ~ (c 0.38 in CHC13); Aizal. Found: C, G2.34; H, 6 . i 4 ) are in harmony with those reOXIDATION OF STEROIDS BY MICROORGANISMS. ported by Bernstein, et al.' Oxidation of the diacetates of I11 and I V with IV. 16a-HYDROXYLATION OF 9a-FLUOROHYDROCORTISONE AND 9a-FLUOROPREDNISOLONE BY chromic and sulfuric acids in acetone gave, respecStrepfomyces roseochromogenus tively, the diacetates of 16a-hydroxy-9a-fluorocorSir : tisone, m.p. 213-217'; [ a I z 3 +94" ~ (c 0.35 in 235 mp (15,100); A?:' 2.M, The introduction of a 1Ga-hydroxyl group into CHCl,); &A: 0a-fluorohydrocortisone (I) and Sa-fluoropredniso- 5.75, G.00, (5.19p ; Anal. Found: C , 63.03; H, (j.53, lone (11) has been shown by Bernstein, et aZ.,' to re- and of 1G a-hydroxy-9a-fluoroprednisone, 111.p. 205[a]"D +go" (c 0.41 in CHC1,); A:!& za3 sult in complete suppression of the salt-retaining 206'; 2.9i, 5.i5, 5.09, 6.15, G.22, properties of these steroids without appreciably mp (lG,200); A%?' impairing their glucocorticoid activity. More- 11.28p ; Anal. Found: 63.13; H, 6.23. The biosynthetic conversion of I11 into I V by over, preliminary studies in man2 have demonstrated the anti-arthritic activity of Sa-fluoro-1Ga- means of Corynebacterium simplex has been rehydroxyprednisolone (triamcinolone) and have p0rted.l \Ye have performed this reaction in 65 confirmed its lack of salt-retaining activity. An yield with a strain of Jfycobacterirrna rliodocliroii eKicient synthesis of this complex steroid is there- adding the steroid (0.5 g. / l . ) t o a 24-hour old culture in a medium containing yeast extract, tryptone, pcnfore of considerable practical interest. tane, glucose calcium, and incubati CHzOH I
C=O
\/:\/ 1 :
IY,S
=
O H ; AI
*
o,,~'\." Alpplyingthe microbiological 1Ga-hydroxylation reaction first reported from this laboratory3b4t o I arid I1 we have succeeded in preparing 1Ga-hydrosy-9a-fluorohydrocortisone (111) and 1Ga-hyc!roxy-9 a-fluoroprednisolone (IV) in yields of 50'/c and 200c, respectively. -1 fermentation medium containing soybean meal (30 g.), glucose (20 g.), soybean oil (4.4 g.) and calcium carbonate (0.050 g.) in distilled water (2 1.) was distributed over 40 250-ml. erlenmeyer flasks, steam-sterilized for 30 minutes a t 120' and after addition of the steroid (1 g. dissolved or suspended in 40 ml. of methanol) inoculated with vegetative growth of Streptomyces roseochromo(1) S. Bernstein, R . H. L e n h a r d , W. S. Allen, M. Heller, R . Littell, S. id. Stolar, L . I. Feldman, and R . H. Blank, THISJ O U R N A L , 78, 5693
( I iizii:. ( 2 ) L. Hellman, B. Zumoff, RE. K. Schmartz, T. F. Gallagher. C . A. Berntsen a n d R. H. Freyberg, Abstract of Papers presented a t t h e 3 r d Interim Meeting of t h e American Rheumatism Sssociation, Bethesda, M d . , Nov. 30, 1956. ( 3 ) D . Perlman, R. 0 . T i t u s and J . Fried, 'THIS JOT:RNAL, 7 4 , 3126 (1952). (4) J . Fried, R. W. Thorna, D . Perlman, J. E.: Herz, and .A Rnrmnn, R . r i ' i : f P m w . ITo~niriir Rpvr.oTrh, 9, 140 (1955).
( 5 ) TVaksman Collectirrn Numher 3689, Iln Rriinsn-ick, h-.J . (U) Occasionally a polymorphic f o r m of 111 \ v a s ohtaincrl, n h i c h lacked t h e 2.79 band and showed an entirely different picture in t h e fingerprint region ( 7 ) T h i s culture, isolated b y Dr. J . 0. Lnmpen of our laboratories, is identified as SC 2318 i n our collection. W'ith regard t o its classification cf, R . Gordon a n d J M. X i h m , J Rncl., 73, 15 (1957). I t s dehydrogenating prriperties were discovered in these laboratories I,? Dr. H. Kroll.
THE SQUIBB IXSTITUTE FOR
KICH.4RD 1%'. THOMA JOSEP FRIED
MEDICAL RESEARCH SALTATORE Bosa~so PATU, GRAIKI\VICA NEWRRUNSIVICK,hT.J. RECEIVED JULY 31, 19~57'
TOTAL SYNTHESIS OF
17-DESMETHOXYDESERPIDINE
Sir: I n a previous communication' we described the synthesis of the unsaturated lactone (I). \Ye now wish to report the conversion of this intermediate to 17-desmethoxydeserpidine. Reduction of the unsaturated lactone (I) with hydrogen and a highly active platinum catalyst for sixteen hours gave predominantly the saturated lactone (11),m.p. 287-269" (found: C, 74.81; H, 6.98; AN"l0l 5.71 p) as well as a small amount of an isomeric lactone (111), m.p. 30'7-309' (found: C. 74.25; H . 6.86; X%:F1 5 . G p). When the product was allowed t o remain in contact with the (1) F I. Weismhorn a n d 11 I: 20021 (19.56)
Applegate, THIS Tlirrsu 11
78,
