April 20, 1957
COMMUNICATIONS TO THE EDITOR
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These compounds exhibit an order of inhibition for 7 days, harvested and extracted.' The exof carbonic anhydrase previously observed only tract was saponified, chromatographed and the among the heterocyclic sulfonamides and in carotene crystallized without carrier, with yields animals promote the renal excretion of sodium. 7 and 9 mg., respectively. Both were recrystalI n addition, however, they produce also a marked lized with resulting yields of 5.8 and 6.0 mg. The activities of medium and carotene were deincrease in chloride excretion and cause a diuresis not unlike that observed with organic mercurial termined in triplicate. Samples were burned in a compounds. One compound of this type, 6-chloro- Pregl furnace and the C02 was precipitated from 7 - sulfamyl - 1,2,4- benzothiadiazine-l,1 -dioxide (11, sodium hydroxide as BaC03. The carbonate was R = H) has been selected for clinical trial and plated on 1-inch copper planchets and counted in a assigned the generic name of chlorothiazide. gas-flow counter. Counts were recorded on a Preliminary results3 in man substantiate the im- Tracer-Lab Autoscaler. Activities were corrected portant pharmacological findings observed in our for self-absorption. The presence of leucine in the culture medium laboratories. Chlorosulfonation of m-chloroaniline a t 150' in visibly stimulates production of @-caroteneby the the presence of sodium chloride4yielded 6-amino-4- mold, but the incorporation of leucine carbon into chlorobenzene-l,3-disulfonylchloride (111) which carotene is insignificant for 1-C and limited for with ammonium hydroxide gave 6-amino-4-chloro- 2-C.' The loss of the carboxyl carbon was to be benzene-1,3-disulfonamide (IV) (m.p. 251-252' ; expected, but the limited incorporation of 2-C Anal. Calcd. for CsHsClN304S~: C, 25.22; H, made it impossible for the leucine to provide the 2.82; N, 14.71. Found: C, 25.48; H I 2.81; N, whole of the requisite Cb repeating unit in the caro14.68). Reaction of 111 with acetic anhydride tene molecule. afforded 6-acetylamino-4-chlorobenzene-l,3-disul~- From media in which 6.4y0 of the total carbon is fonyl chloride (m.p. 137-139'; Anal. Calcd. for represented by leucine carbon, we find an enrichC8He,C&NO&: C, 26.21; H, 1.65; N, 3.82. ment of 3.6-fold for the 3-C, and of 8.7 for 4-C, in Found: C, 26.39; H, 1.77; N, 3.79) which with the carotene. The specific activities of the leualcoholic ammonia gave a mixture of 6-acetyl- cines and carotenes are shown below, in cts. per amino-4-chlorobenzene-l,3-disulfonamide (I, R = min. per mg. BaC03, together with the activities CH3) (m.p. 261-262' dec.; Anal. Calcd. for CS- of the media determined by independent combusHl&1N30&: C, 29.31; H, 3.08; N, 12.82.. tion and by calculation based on combustion of the Found: C, 29.49; H , 3.25; N, 12.85) and 6-chlo- leucine alone. The activities are the averages for l-, r0-3-methyl-7-sulfamyl- 1,2,4-benzothiadiazine-1, Specific activity 3-Cl4-Leucine 4-Cl4-Leucine dioxide (11, R = CH3) (m.p. 332' dec.; Anal. Calcd. for C8HsClN304S2: C, 31.02; H, 2.60; N, Leucine 1442 2350 13.57. Found: C, 31.27; H, 2.53; N, 13.50) that Medium calcd. 94.0 149 was separated by recrystallization from water. Medium exptl. 96.8 159 When heated with formic acid under reflux, IV was Carotene 343 1392 converted to 6-chloro-7-sulfamyl-l12,4-benzothiadiazine-1,l-dioxide (m.p. 342.5-343' dec.; Anal. the six carbons of leucine and for the forty of caroCalcd. for C ~ H ~ C ~ N ~ O C,~ 28.43; S Z : H , 2.05; N, tene. The true activities of carbons 3 and 4 are 14.21. Found: C, 28.65; H, 2.23; N, 14.11). six times greater and the extent to which they are incorporated is given by the expressions 343/1442 (3) R . V. Ford and C. L. Spurr, "Electrolyte Excretion Patterns X 40/6 or 1.59, and 1392/2350 X 40/6 or 3.96, Due to Chlorothiazide, A New Orally Effective Diuretic Agent," Presented a t the Southern Society for Clinical Research, h-ew Orleans, respectively. January 2F, 1957. Up to this point, we have introduced no assump(4) 0. Lustig and E. Katscher. Monotsh., 48, 87 (1927). tions. In all our work, the primary carbon source MERCK,SHARP AND DOHMERESEARCH LABORATORIES has been glucose which comprizes ca. 90% of the DIVISIONOF MERCKA N D Co., Ixc. carbon of the medium. With labeled glucose, the WESTPOINT,PENNSYLVAXIA FREDERICK C. NOVELLO JAMES M. SPRAGUEspecific activity of the carotene is less than that of the glucose, and the extent to which the glucose carRECEIVED FEBRCARY 18, 1957 bon is discriminated against varies with the nonglucose carbon source, e.g., yeast autolysate, glycine.3 If the assumption is made that formation of caroCAROTENE, 3-C1*- AND 4-C1'-LEUCINE tene derivable from non-leucine sources is unafSir : 3-(2'"- and 4-C14-d,l-leucines were synthesized fected by the leucine, the labeled carotene derived by conventional methods from carboxyl labeled from leucine has been diluted by an equal quantity isobutyric acid and carbonyl labeled acetone, re- of inert endogenous carotene. The figures 1.59 and 3.96 may therefore be mulspectively, and incorporated into culture media in which Phycomyces blukesleeanus was grown. The tiplied by a factor of ca. two. This makes i t posculture medium consisted of glucose (2.5% w./v.), sible for the 4-C of leucine to appear eight times in asparagine (0.i25Y0), and 3-C14-leucine or 4-C14(1) C. 0. Chichester, T. Nakayama, G. Mackinney and T. W. Goodleucine (O.125y0), together with inorganic salts win, J . Biol. Chcm., 314, 515 (1955). C. 0. Chichester, P. S. W o n g and G. Mackinney, Plant Physiol., and thiamine.' Thirty petri dishes were inocu- 29,(2) 238 (1954). lated with an active spore culture of P . bluskesleea(3) C. 0. Chichester, T. Nakayama and G. Mackinney, J . B i d . nus. Cultures were grown a t room temperature c i ~ m . a21, , 819 (1956).
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BOOK REVIEWS
the carotene molecule, and the position of this key carbon in the Cg repeating unit of carotene can readily be envisaged. Whether leucine provides an is0 C B fragment cannot be determined as yet, but it is clear that the stimulatory effect of leuciue on carotene formation is due to the incorporation of leucine fragments into the molecule.
Vol. 7!1
A grant from the National Science l'uuiidation is gratefully acknowledged. DEPARTMENT OF FOODTECHXOLOGY €1. I'OKOYA\IA UNIVERSITYOF CALIFORNIA
e. 0 . CHICHTSTER
A N D BERKELEY*
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T.
NAKAYAMA
A. LUKTON G. MACKINNLY
RLCLIVLD M A R C H11, 1957
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BOOK REVIEWS Heterogeneous Equilibria and Phase Diagrams; Solutiolis Synthetic Polypeptides. Preparation, Structure, and Properties. Volume V. B Y C . H . BAMFORD,A. ELLIOTT of Konelectrolytes; Statistical Mechanics; Radiation and W. E. HANBY, Courtaulds Ltd., Research Laboratory, Chemistry; Quantum Theory; Ion Exchange; Polymers; Maidenhead, Berkshire, England. Physical Chemistry. Kinetics of Reactions in Solution; Kiuetics of Reactions in A Series of Monographs. Edited by Eric Hutchinson. Cases; Combustion and Flames; High Temperature Academic Press, Inc., Publishers, 111 Fifth Avenue, Chemistry; Thermochemistry and Thermodynamics o f New York 3, N.Y. 1956. xiii 445 pp. 16 X 23.5 Substances; T h e Solid State; Isotopes; Magnetic Kesociii. Price, $10.00. nance; Surface Chemistry and Catalysis; Molecular Electronic Spectroscopy; Vibration-llotdtion Spectroscopy; ExThe special interest in the synthetic polypeptides lies riot perimental Molecular Structure. only in their relation to proteins but in the fact that they The literature coverage is for the year 1055. Because { i f constitute a class of high polymers exhibiting with unusual space limitations and other reasons, none of t h e authors clarity t h e relation between properties and molecular claim complete coverage of the literaturc. Even with such structure. I n general this book can be read by all who have a basic restrictions almost 3300 references are cited. In some of the topics the coverage is more complete than in others. interest and training in t h e natural sciences and by all who In the article on quantum theory t h e reviewers liniitctl are concerned with high polymers. The polymers which are the subject of this book are the themselves to a timely and pertinent survcg of crystal fieltl synthetic linear polypeptides derived from amino acids. theory. Their discussions mainly concerned t h e transition iiietal complexes. Such limitations are not serious an(1 I r i the earlier chapters difficulties in the preparation arid probably are desirable when one considers the matter frciiii study of the polypeptides are discussed together with the the point of view of the whole series of reviews, since in preparation froin various beginning materials and the beprevious years other topics in quantum theory have been liavior of the final products toward different solvents. Chapter I V has to do with chain configurations in the adequately covered. The s t y k and method of preseritatiriii of the reviewers of course vary from individual t o individual polypeptides, bond lengths and angles, folding of chains, but on the whole the material is well presented. The hydrogen bonding, etc. Chapters V and VI are concerned with Infrared Spectros- editors and authors are t o br complimented on a job well done. This review should be on the shelves of all scientists copy and the effect of various agents (Hydrogen bonds, interested in physical chemistry. Deuterium) on this method of examination. Chapters V I I , VI11 and IX deal with X-ray studies on I~EPARTMENT OF CHEMISTRY various types of polypeptides. \V,lLTER s. KOSKI JOHXS HOPIEINST;NIVERSI~TY In Chapter X are discussed the properties of synthetic THE polypeptides-molecular weights, carboxylic acid effects, BALTIMORE 18, R I A R Y L A N D solubilities of various types, dyeing properties, optical rotation and molecular orientation. Medicinal Chemistry. Volume 111. A Series of Reviews Chapter X I is a brief review of the biological properties Prepared under the Auspices of the American Chemical of the synthetic polypeptides-enzymic hydrolysis, jnterSociety. Editors: F. F. BLICKEand R . H. COX. Authors: action with viruses and bacteria and blood clotting. T. P. Carney, P. I,. deBenneville, V. Papesch, E. 1'. The final chapter takes up the fibrous proteins iii their Schroeder, A. Stempel, and J. 2. Aeschlimann. John relation to the synthetic polypeptides, the silks, the kcratins IViley and Sons,Inc., 440 Fourth Avenue, New York 16, of wool and hair, their amino acid residues with their N.Y. 1956. vi 346 pp. 15.5 X 23.5 cm. Price, sequence and the physical structure as revealed by X-ray 510.50. : ~ n dinfrared observations. There is a 16 page appendix wliich includes the discussion The four review monographs dealing with the subject\ of Dichroisni in partially oriented polymers, use of atomic Methadone and Related Analgesics, Quaternary Anlmoniuiri models, etc. Finally there is an excellent subject index of Germicides, Non-mercurial Diuretics, and Synthetic Arianineteen pages. Iogues of Physostigmine are written by experts in their ficltl for experts. This volume, like its precursors, should be of DEPARTMENT OP BIOCHEMISTRY iiiterest not only to the synthetic organic chemist, but also UNIVERSITY O F ROCHESTER SCIIOOI, OF the pharmacologist. The mass of detail is, for the most MEDICINEA N D DENTISTRY \v. R . II1,OOR to part, skillfully handled, with concise discussions of the tcxt ROCHESTER, NEW YORK upon which the special reader may amplify through rcference t o one of t h e numerous citations of the original literature. T h e task of thoroughly covering many homolAnnual Review of Physical Chemistry. Voluliie 7. I I . ogous series of compounds is skillfully handled by the use EYRING,Editor, University of Utah, C. J. CHRIsrEssEx, of detailed tables containing original literature references. Associate Editor, IJniversity of Utah, and H. S. JOHNSTON, l h e format, type style and freedom from errors are Associate Editor, Stanford University. Aniiual Revimvs, riotable. In t h e last connection, only a few typographical Inc., Palo Alto, Stanford, California. 1956. vii 50:3 errors were encountered. lip. 16 X 23 cin. Price, $7.00. In these tiuies of tremendous incrciise ill c o l u t i i c o f ~,iNIVERISrYO F ROCHESTER scientific literature, Volunie 7 of the Artiiual Review of SCHOOL OF MEDICINEA K D DNNUSIRY Physical Chemistry is very welcome even to a specialist i l l D O X 287, STA. ?: I.LON I. ~ l I l ~ l , I m a field. T h e topics covered in this edition are: Cryogenics; ROCHESTER 20, NEW YORK
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