Aug. 5, 1954
COMMUNICATIONS TO THE EDITOR
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droxyl groups into the substrate has been ob- Pratt, Jennie I. Mejeur and Henrietta Triemstra served.1~2~5~6~12 However, dihydroxylated products and Mr. J. R. Heald and G. Staffen for technical have been formed to any appreciable extent only assistance, to Dr. J. L. Johnson and associates for the spectra and to Mr. W. A. Struck and associwith p r o g e ~ t e r o n e ~or * ~ 3-ketobisnor-4-cholen.~~~~ ates for rotations and microanalyses. 22-all as a substrate. Since thus far no 17-hydroxylation by microbioP. D. MEISTER L. M. REINEKE logical means has been reported, i t is the purpose of LABORATORIES R. C. MEEKS this communication to describe the enzymatic RESEARCH H. C. MURRAY THEUPJOHNCOMPANY oxygenation of the 17a-position of Czl-steroids by KALAMAZOO, MICHIGAN S. H. EPPSTEIN Cephalothecium roseum Cda (A.T.C.C. 8685). Upon H. M. LEIGHOSBORN incubation of 11-desoxysteroids with C. roseum, A. WEINTRAUB D. H. PETERSON the 17a-hydroxyl group was introduced with or RECEIVED JULY 6, 1954 without concomitant hydroxylation of the Sp- or the 1la-position. When 11-keto or 1lp-hydroxy17-desoxysteroids were used as substrates only 17THE STEREOCHEMISTRY OF METAL hydroxylation took place. CHELATES OF A POLYDENTATE LIGAND When desoxycorticosterone was incubated with Sir: a 48-hour growth of C. roseum on a corn steep-gluMetal complexes with sexadentate ligands were cose medium for 48 hours, extraction of the fermentation liquor with methylene dichloride and chroma- first described by Dwyer and Li0ns.l Diehl and tography of the concentrates over Florisi13 yielded co-workers2 seemingly prepared some similar comtwo products: (a) 1la,l7a,21-Trihydroxy-4-preg-pounds with triethylenetetramine and salicylaldenene-3,2O-dione2t4t6+l4 (11-epi F). The identity of hyde (also substituted salicylaldehydes) by the this compound was established through these physi- reaction of cobalt salts during their investigation on cal constants: m.p. 206-211 O, m.p. in admixture with the oxygen-carrying synthetic chelates. They, howauthentic 11-epi F, 208-212'; [ a ] =+121" (metha- ever, did not isolate the pure compounds or study nol); ( [ a ] =+117O in methanol for authentic 11-epi their properties. A Schiff base (I)from triethyleneF); the infrared spectrum of the diacetate (in HOC6HaCHN(CH2)zNCHzCHzN(CHz)&:=CHCeH4OH chloroform solution) was identical to the spectrum \CH/ of the authentic 11-epi F diacetate. (b) 6P,17a,I 1 21-Trihydro~y-4-pregnene-3,20-dione~~~~~~ was also CsH4OH isolated; m.p. 234-236', [ a ] f~5 3 " ; A&. 238mp, E 12,900. Anal. Calcd. for CalHaoOs: C, 69.58; tetramine (11) and salicylaldehyde (111) has been ,~ gives a brown hyH, 8.34. Found: C, 69.58; H, 8.38. The infrared reported by M ~ k h e r j e e which spectrum of the diacetate, m.p. 190-193', was iden- groscopic substance with [Co(py)*Cl*]CI. This is soluble in water, insoluble in alcohol, and decomtical to the spectrum of an authentic specimen. Paper chromatography indicated that small poses a t 110'. The same substance is also said to amounts of Reichstein's compound S and epicorti- be obtained by the reaction of (111) on [Co trien C12]Cl. costerone had been formed. The nitrogen analog of Dwyer's Schiff base (IV) Fermentation of the following substrates with C. roseum gave the corresponding 17a-hydroxy HOCsH4CH=N( CHz)zNH(CHz)zNH(CHz)zN=CHCaH4OH derivatives : could not be prepared from (11) and (111),only (I) being obtained. We have, however, been able to Substrate Conversion Product Progesterone llor,l7or - Dihydroxyprogest- prepare a large number of compounds of the sexadentate ligand (IV) by the action of (11)on bis-salierone cylaldehyde compounds of cobalt, copper and other 11-Dehydrocorticosterone Kendall's Compound E Corticosterone Kendall's Compounds F and bivalent metals in about 80-85% yield. These can be prepared also by adding a mixture of (11) E and (111) in four equivalents of alkali (KOH) in a It is worthy of note that the enzyme systems of cold methanol solution to a solution of the metal salt Cephalothecium can introduce a hydroxyl group a t concerned. (I) also gives a poor yield (10-20%) carbon atom 17 without any interference from a of these compounds by careful manipulation. A hydroxyl group already present a t carbon atom 21 compound of the composition [Co trien CHOCa(as in desoxycorticosterone). This is in contrast t o H40]++ is obtained as the main product from [Co the performance of the mammalian adrenal which, trien C12]Cland (111). a t least under the in vitro conditions employed by When base (I) reacts with metal salts, one molethe Worcester group, l6 cannot oxygenate the 17- cule of salicylaldehyde is hydrolyzed from it, and position of desoxycorticosterone. derivatives of base (IV) result. For bivalent metal The authors are indebted to Misses Irene N. ions such as Fe'I and PdlI these have the composition intermediate between MTS2 and MTS3, (14) For chemical syntheses of this compound or its diacetate, cf. J. Romo, et al., Chcm. and Ind.. 783 (1952); H. L. Herzog, et al., THIS where TSP and TS3 represent the bi-negative ions JOURNAL, 74, 4470 (1952); A. Lardon and T. Reichstein, Phorm. of the bases (IV) and (I), respectively. Acta Hclu., 47, 287 (1952); J. Romo, e1 al., THISJOURNAL, 76, 1277 The complexes of (IV) will be asymmetric (1953). (16) For a chemical synthesis of this compound cf. I(.Florey end M. Ehrenstein, J . Org. Ckcm., in press. (In) H.T.evu, el a t , , I,Blal. Chrm., 108,4118 (196S),
(1) F. P. Dwyer and F. Lions, THISJOURNAL, 69,2917 (1947). (2) H. Diehl a n d co-workers, Iowa Slat# COIL1.Sci., 91, 108 (1947). (8) A. K.Mukbcrjee, Scicncr nnd CuNwrr, 107 (186s).
le,
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COMMUNICATIONS TO THE EDITOR
whether the base behaves as a tetradentate or hexadentate ligand. When (IV) behaves as a tetradentate ligand (as in the copper and palladium compounds obtained by the first two methods of preparation), the metal complexes have an asymmetric center in the metal ion, due to the two aromatic rings lying in different planes, irrespective of whether the arrangement be planar or tetrahedral. There is of course a possible meso-form. Some of the compounds prepared are tabulated: Compound
[CO"TSs] .0.5H20 [Co"'TS2] C1.2.5H20 [Fe1"TS2]I.1.BH20 [ Cu"TS21
[Al"'TS2] 1.0.5H20
Color and crystal form
Brownish yellow powder Dark black-brown hexagonal plates Dark purple rectangular prisms Bluish green prisniatic needles, green when anhydrous Colorless, hexagonal
lyzed against buffers of different FH, all a t 0.1 ionic strength, and were then ultracentrifuged a t about 2.5'. Between pH 7.5 and 4.6, the ultracentrifuge diagrams were essentially unchanged, closely resembling those of Fig. 2c of reference (1). From pH 4.6 to 3.1 progressively larger amounts of 3 component with sedimentation rate corresponding to free antibody y-globulin appeared in the diagrams, while the peaks due to complexes diminished in area. At pH 2.4, the diagram was that of a corSolubility Water E t O H
Magnetic moment
Melting point, OC.
insol.
sol.
4.37
dec. 226
sol.
sol.
0
210
sol.
sol.
1.83
117-118
sol.
sol.
2.01
78
sol.
sol.
...
