Sept. 5, 1960
COMMUNICATIONS TO THE EDITOR
temperature,6 we suggest t h a t in dilute solution and in the molten state the isotactic and block polymers of optically active a-olefins are a t least in part spiralized and that helices of a single screw sense largely prevail. ( G ) For t h e highly stereoregular fractions of poly-(S)-3-methyl-lpentene which have a very low solubility and high melting point it is possible t h a t t h e solutions of t h e polymers still cuntain crystalline molecular aggregates which undergo dissociation by increasing t h e temperature. I n this case, t h e decrease of the optical activity by increasing the temperature, could be attributed t o the dissociation of the crystalline aggregates ha\.ing high optical activity t o dissolved less optically active macromolecules, which may change or eventuall) loose their spiralized conformation. G. S a t t a . RI. Farina, RI. Peraldo, P. Corradini, G. Bressan a n d P. Ganis (Reird. Acc. h ' n s . L i n c e i , April, 1960) have found crystalline molecular aggregates in solutions of some di-isotactic polymers. We are particularly indebted t o Prof. N a t t a and his co-workers for t h e discussion on this point. ISTITUTO D I CHIMICA ORGASICA ISDUSTRIALE UNIVERSITA DI PISA
VIA RISORGIMENTO 19 PISA,ITALY RECEIVED JUNE 4, 1960
P.P I N 0 G. P. LORENZI
4747
III'O and the hitherto unknown amino acid IV designated by us as thiostreptoic acid. NH2 S-CH I I I/ CHZ-CH-Cb /C-COOH
S-CH I I R-C+ /C-COOH
i
h'
N CH,-CH-W 'C-COOH I I NH: S-iH
N I, R = CH2NH2 11,R CH&H*CHNH, 111,R=CH,CH:CO
IV
S-CH I I1 CH?-CO-Cb /C-COOH
N N\C-COOH CH9-CO-CP I I/
I
S-CH
v
The crystalline acid IV was isolated from the hydrolysate after removal of impurities by butanol DEGRADATION OF THIOSTREPTON. extraction. The dihydrate melts a t 235-237" THIOSTREPTOIC ACID (dec.); * O (c, 1.0 in 1 N HC1); XitxHc' Sir: 236 rnp ( e = 15,000); Anal. weight loss a t 110": Earlier publications from this Laboratory have 9.7; calcd. for 2 H 2 0 : 9.5; calcd. for C12H1404N4S2: described the isolation and characterization1 as C, 42.1; H , 4.12; N , 16.4; S, 18.17. Found: C, well as the biological proper tie^^,^ of the antibiotic 42.3; H, 4.21; N , 16.4; S, 18.6. On Whatman thiostrepton. Kenner, et u Z . , ~ have isolated the No. 1 paper in a system of 1-butanol-acetic acid4-thiazolecarboxylic acids I and I1 from acid hy- water (4 : 1: 1) thiostreptoic acid moves somewhat drolysates of the antibiotic. This Communication faster than cystine, Rf = 0.08-0.10. In 1 N presents our own degradative studies with thio- HCl it forms a crystalline dihydrochloride, which, strep-ton. when dissolved in water, deposits the free acid. Separation of water-soluble components of acid Treatment of IV in water with acetic anhydride in hydrolysates (6 NHC1,lG hours a t 105') by counter- the presence of triethylamine yields the correspondcurrent distribution, then partition chromatog- ing N,N'-diacetyl derivative, m.p. 275-277' (dec.). raphy, led to the isolation of L-threonine, L- Anal. Weight loss a t l l O a , 10.4; calcd. for 3 H20: isoleucine, L-alanine, and of c cystine.^ The iden - 11.2; calcd. for C16H1806N4S2: c, 45.3; H , 4.26; s, tity of these components was ascertained by com- 15.1. Found: C,45.3; H,4.38; S,l5.l. Onbotha parison with authentic samples on paper chromato- reaction with dinitrofluorobenzene I V forms monograms, by infrared spectra and by measurement of and bis-dinitrophenyl derivative." The former their optical rotation. No other conventional amino is ninhydrin positive. From the specific absorpacids were found. From a partial hydrolysate tion of the monodinitrophenyl derivative a t 35,5 (6 N HCI a t room temperature) L-isoleucyl-L- mp a molecular weight of about 500 was calculated. alanine6 was obtained in crystalline form and iden- Titration of monodinitrophenylthiostreptoic acid tified by paper chromatographic separation of the with alkali and acid gave neutralization equivacomponents liberated by hydrolysis before and lents of 2 5 i and 539, respectively (mol. wt., 508). after dinitrophen ylation. Reduction of IV with sodium in liquid ammonia Hydrolysis of thiostrepton with a 1:1 mixture and then acid hydrolysis12 led to a mixture, in of concentrated hydrochloric and formic acids7 which alanine and cystine were identified by paper furnished in addition to the previously isolated4 chromatography. Oxidation of thiostreptoic acid thiazolecarboxylic acidss I and IT,9the keto acid (8) T h e fact t h a t acid hydrolysis of t h e product obtained from thio (1) J. Vandeputte and J . D. Dutcher, "Antibiotics Annual, 19551Y56," Medical Encyclopedia, Inc., New York. N. Y., p . 5 6 0 . (2) J . F. Pagano, M. J . Weinstein, H . A. S t o u t a n d R. Donovick, i b i d . , 1955-1950, p. 554. ( 3 ) B . A. Steinberg, a '. P. Jambor and Lyda 0. Suydam, i b i d . , 19551956, p . 5 6 2 . ( 4 ) G. W. Kenner, R. C. Sheppard a n d C. E. Stehr, Tcbohcdron Lclfcrs, 23 (1960). (5) I n thiostrepton this D-amino acid occurs in t h e reduced form, a s shown b y the absence of a reaction for disulfides in t h e antibiotic and t h e positive test for sulfhydryl after hydrolysis. Moreover, t h e behavior of this amino acid during fractionation by countercurrent distribution or b y partition chromatography was characteristic of cysteine rather t h a n cystine. (6) Isoleucine and alanine were also found in a diketopiperazine formed during pyrolysis of t h e antibiotic a t 250' i n vacuo. (7) G .L. Miller and V. du Vigneaud, J. B i d . Chcm., 118, 101 (1937).
strepton by reduction with sodium in liquid ammonia yields glycine and a-aminohutyric acid indicates t h a t t h e thiazolecarboxylic acids I and 11, and probably also IV, are present a s such in t h e parent molecule and not, for instance, as thiazolines. (9) I n these studies, I 1 was isolated a s a crystalline salt with Phydroxyaeohenzene-p'sulfonic acid. Anal. Calcd. for CioHmOaS&: C,49.1; H , 4.34; S, 13.8. Found: C, 48.9; H, 4.46; S, 13.7. ( I O ) T h e acid 111,probably formed a s a secondaly degradation product from 11, was found t o be identical with t h e acid isolated by P. Brookes, A. T. Fuller and J . Walker (J. Chem. Sac., 689 (1957)) from Micrococcin P . A homologous keto acid was reported a s a secondary degradation product from bacitracin A (J. R. Weisiger, W. Hausmann a n d L. C. Craig, THISJOURNAL, 17, 3123 119551). (11) A. R. Battershy and L. C. Craig, THIS JOURNAL, 1 3 , 1887 (1951); 74, 4023 (1952). (12) P. Brookes, R . J. Clark, B. Majhofer, M. P. V. Mijovic and J. LX'alker, J. Chevi. S o c . , 925 (IQCO).
4748
COMRIUNICXTIONS TO THE EDITOR
Vol. 82
with K M n 0 4 in dilute NaOH13 gave the diketo intermediate produced by a ring cleavage, underacid V, m.p. 280-290' (dec.); X",'ix 293 mM ( E = goes decarboxylation and/or intramolecular cycliza9,000); anal. Calcd. for C ~ ~ H ~ O G N ~C,S Z42.3; : tion. Chemical conversions of benzene derivaH, 2.37; S, 18.8. Found: C , 12.4; H , 2.19; tives to pyridine derivatives by cleavage of the S, 18.3, in addition to a small amount of 4-thiazole- carbocyclic ring and recyclization have not been carboxylic acid. The above data permit the assign- developed. ment of structure IV to thiostreptoic acid. Unambiguous proof was provided by a synthesis of v. Succindialdehyde was treated with potassium cyanide and the resulting cyanhydrin benzoylated I I1 I11 in s i t z ~ . ' ~ , . 'A~s expected, two isomers, the meso form and the racemate of a,G-dibenzoyloxyadipic We wish to report a new and general method for acid dinitrile were isolated, n1.p. 135-137" ; and the chemical conversion of 3-aminocatechols (IV) m.p. 212-216'; anal. Calctl. for C20H1604N2:to 6-hydroxypicolinic acids (VII), Unstable %amiC, 69.0; H, 4.63; N, 8.94. Found: C, GS.9; nocatechol (I\-a), 4 3,4-dihydroxyanthranilicacid H, 4.60; N, 7.93 for the lower melting and C, 69.0; (IVb),5 m.p. 175" (dec.), unstable 2,3-diaminoH , 4.81; N , 8.10 for the higher melting isomer. 4,3-dihydroxytoluene (IVc),e 1,2-dihydroxy-3-amiThe lower melting isomer was used in the subse- nonaphthalene (IVd), m . p 164' (dec.), arid unstable quent steps. Treatment with hydrogen sulfide 1,2-dihyclroxy-3-aniino-4-anilinonaphthalene in dimethylformamide in the presence of triethanol- are transformed respectively to 6-hydroxypicolinic aminel3>l4furnished a,&dibenzoyloxyadipic acid acid (VIIa), m.p. 261-263", 6-hydroxyyuinolinic dithioamide, m.p. 203-205'; anal. Calcd. for acid (VIIb), m.p. 253-2545', 3-axnino-4-methyl-6S, 15.4. Found: N, hydroxypicolinic acid (VIIc), m.p. 285-288' (dec.), C Z O H ~ U O ~ NN ~ ,S ~6.73; : 6.94; S, 15.4, which was condensed with ethyl isocarbostyril-3-carboxylicacid (L'IId), m.p. i11Sbromopyruvate to give the diethyl ester of 2,2'- 320" and I-anilinoisocarbostyril-3-carboxylicacid (1,-klibenzoyloxytetraniethylene) - bis - [I-thiazole- (VIIe), m.p. 250-256O (dec.).8 carboxylic acid], m.p. l 4 4 - 1 G o ; anal. Calcd. Silver oxide in anhydrous ethyl acetate oxidizes for C,,oH280,NaS2:N , 4.60; S, 10.5. Found: N, each dihydroxyamine (IV) to a blue-black unstable 4.68; S, 10.6. Hydrolysis with potash in ethanol aminoquinone (V) .9 In an extension of the applicayielded the corresponding dihydroxy acid, m.p. tion of the Baeyer-Villiger reaction to o-quinones, lo 240-242' ; Anal. Calcd. for C ~ Z H ~ ~ O G N C, Z S ~ each : aminoquinone, after separation from inorganic 41.9; H, 3.51; N, 8.23. Found: C, 42.0; H, material, is oxidized without isolation from solvent 3.05; N, 5.35, which on oxidation with NazCrzOi with a peroxy organic acid apparently to an ungave 2,2'-succinylbis- (I-thiazolecarboxylic acid), isolated derivative of muconic acid anhydride (VI). m.p. 284-2S6" (dec.); A"':, 5.84 sh, 5.90; anal. On treatment of the anhydride with water a subFound: C, 42.5; H , 2.90, identical in all respects stituted muconic acid presumably is formed. with the diketo acid V derived from thiostreptoic Hydrolysis of monocyclic muconic acid anhydrides acid. in the presence of strong ultraviolet light gives only The sulfur content of the antibiotic (9.4%) con- intractable tars. Apparently cis-cis muconic acid sidered in conjunction with the four sulfur-contain- derivatives are produced initially and isomerize to ing fragments isolated so far require for thiostrepton trans-trans modifications.'l In the absence of a minimum molecular weight of 1700.15
(2) H. S. Mason, "Mechanisms of Oxygen Xletabolism" in "Advances in Enzymology," Vol. 19, Interscience Publishers, I n c . , S e w York, N. Y., 1957, p. 92. (3) Ozonolysis of a substituted aminomethylcatechol ether and then cyclization accounts for t h e construction of ring I11 as a pyridone in t h e synthesis of strychnine (R. B. Woodward, M. P Cava, W. r). Ollis, -4.Hunger, H. U. Daenker and K . Scheinker, T H I S JOURKAL 76, 4749 (1954)). Pyridine is reported t o be one of t h e products from benzene and active nitrogen (P. XI. Aronovich, N. K. Bel'skii, and THESQUIBBINSTITUTE MIKLOSBODASSZKY B. hl. hlikhailov, Izeesf. A k a d . L - a ~ h S.S.S.R., Oldel. Khim. NalLk, FOR hfEDICAL RESEARCH JOHS TIMOTHY SHEEHAN 696 (1956), C . A . 51, 1893 (1957)). (4) Identified as its hydrochloride, m.p. 196-202' (dec.). SEK BRUSSWICK, XEWJERSEY JOSEFFRIED ( 3 ) At one time this compound was considered t o be a n enzymatic NINAJ. WILLIAMS CAROLYN A. BIRKHIMER intermediate in the conversion of I t o 111 (K. Makino, F. Itoh and K. S i s h i , .Yaluue, 167, 1 1 2 (1951)). RECEIVED JULY 28, 1960 (6) Isolated a s its dihydrobromide salt, m.p. 162-176' (dec.) (7) Identified as its hydrochloride salt, m . p . 211-213O (dec.). ( 8 ) Satisfactory analytical d a t a have been obtained for all comCONVERSION OF 3-AMINOCATECHOLS TO pounds and/or their derivatives reported in this Communication. 6-HYDROXYPICOLINIC ACIDS' (9) Through condensation with +phenylenediamine each aminoquinone is transformed into the corresponding derivative of l-aminophenazine which was analyzed. Arninoquinones, tautomeric hyIn the enzymatic conversion of 3-hydroxyanthrs- dri,xyquinrine-irionoimines nnd imino dihydroquinones are regarded as nilic acid (I) to quinolinic (111, K = R ' = C O J I ) , eciuivalent structures f-ir the present purpose. = C0.H) and picolinic nicotinic (111, R = H, (10) P. Karrer. R . Schwyzer and A . S e u w i r t h , Helv. Chim. A c t a , 31, acid (111, R = C02H, R' = H ) , a-amino-&car- 1210 (1948). I n boiling :rater exposed t o an ultraviolet lamp, cis,cis-muconic boxymuconic acid semialdehyde (11) an established acidi l lis) changed quantitatively t o t h e tvnss,luans-isomer (J. A . Elvidge, R . P. I i n s t e a d , B H. Orkin, P. Simn. W. Raer and 17. R , Pattison, J . (1) Financial assistance from Sational Institutes of Health Grants p i o s . H-1295 and C Y , 2 8 9 5 is gratefully acknowledged. Cheiii. SOC. 2225 (1020)). (13) P. Brookes, A. T. Fuller and J. Walker, J . Chein. Soc. 689 (1957). (14) J. F. Olin and T. B. Johnson, Rcc. Tyau. Chim., 60, 72 (1931). (15) Ether extraction of acid hydrolysates (1 S HCI, 24 hours, a t 105') furnished a ycllow volatile crystalline compound. .4?zol. C1?HsOaN: C , 6 2 . 6 ; H, 4.3; S , 6.14; A m e x 265 mp ( c , 25.000), 375 mp (I, 2,000). An additional fragment from t h e same hydrolysate contains alanine acylated by the yellow chromophore.
5%:
R'
,
