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 reported by Bernstein, et al.' OXIDATION OF STEROIDS BY MICROORGANISMS. 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,
Sept. ,5, 1037
COMMUNIC.\TIONS TO THE
platinum catalyst for five days after reduction was complete, an equilibrium was reached in which the initially formed lactone (11) containing a C3 ahydrogen had been transformed almost completely into its more stable C3 P-hydrogen isomer (III).* The lactones (I1 and 111) were shown to be CSepimers possessing the same configurations a t (215, C16 and C ~as O a-yohimbine3 by transformation of both (IVc). compounds to d,Z-17-desoxy-a-yohimbine Thus, hydrogen bromide in acetic acid at 100' converted the two isomers to the same bromo acid (IVa), isolated as the hydrobromide, m.p. 278-279" (found: C, 49.90; H, 4.97; Br, 32.78). Compound
I
120
,
>/A
' I 1y
I
o=c-0
111
o=c-0
l8
v
/v\
CHzOOC OR Va, R = H Vb, R OC(CsH?)(OCH3)8
I1
o=c-0
/V\
