,TIONS COMMUNICA
Sept. 20, 1054
TO THE
EDITOR
4751
phenylalanyl-L-glutaminyl-L-asparaginyl3 - benzylL-cysteinyl-L-prolyl-L-arginylglycinamide(11) , debenzylation of this compound with sodium in liquid ammonia and cyclization of the resulting sulfhydryl nonapeptide to the disulfide. Synthesis of I1 was accomplished by coupling N-carbobenzoxyS-benzyl-~-cysteinyl-~tyrosyl- phenylal alanyl-^XIV xv glutaminyl-L-asparagine (111) with S-benzyl-L-cysbinol (XV) (hydrochloride, m.p. 195-205O dec., teinyl-L-prolyl-L-arginylglycinamide monohydrocalcd. for CzlHz~OZNzC1.2Hz0: C, 61.97; H, 6.69; bromide (IV) . CsHiOH CsHs N, 6.89. Found: C, 61.87; H, 6.97; N, 6.97), I I which was rearranged by hydrogen bromide in NHz 0 CHz 0 CHz acetic acid, followed by boiling aqueous sulfuric I II I It I acid, to isostrychnine I (XVI)2 (m.p. [evacuated CHz-CH-C-NH-CH-C-NH-CH I I tube] 229-230°, [ c Y ] ~ ~ +23 D =k 4" (c = 2.54 c=o [EtOH])), identical in all respects with an authen- S I I 0 iVH
0
S
I
II
CH2-CH-NH-C-CH-NH-C-CH-( I I
I c=o
I CHzN
I
I
XVI
-
tic sample (m.p. [evacuated tube] 228-230°, [CY]*~D $25 f 4" ( c = 2.44 [EtOH])), and further characterized through isomerization by ethanolic potash3 to strychnine (I), identical in infrared spectrum, melting point, and chromatographic behavior with the natural alkaloid.
I
CHz)z-CONHz
CHr
I
COXHz 0
0 II \ " It ,CH-C-NH-CH-C-NH-CHZ--CONH~
I I
CHz-CHz
OH
It
CHZ CHzCHtNH-C-NHz
I
/I
NH
~-Phenylalanyl-~-glutaminyl-~-asparagine~ was coupled with N-carbobenzoxy-S-benzyl-L-cysteinylt tyrosine^ by the isobutyl chlorocarbonate mixed R. B. WOODWARDanhydride procedure5 to give 111, m.p. 208-209", CONVERSE MEMORIAL LABORATORY MICHAELP. CAVA [CY]"D -26" (c 1, dimethylformamide) (calcd. for HARVARD UNIVERSITY W. D. OLLIS~ CAMBRIDGE 38, MASSACHUSETTS A . HUNGER C45H~1011N7S:C, 60.2; H, 5.72; N, 10.9; S,3.57. H . U. DAENIKER Found: C, 59.9; H, 5.77; N, 10.4; S,3.57). K. SCHENKER N"-p- Nitrobenzyloxycarbonyl-L-arginylglycinRECEIVED AUGUST23, 1954 amide was prepared by the procedure of Gish and Carpenter6 and isolated as the picrate (V), m.p. (2) H. Leuchs and H. Schulte, Bcr.. 76, 1522 (1942). 168-171", [CYIz2D-3.8" (c 1, acetone-water ( 4 : l ) ) (3) V. Prelog, J. Battegay and W. I . Taylor, Helv. Chim. Acta, S1, 2244 (1948). (calcd. for cl6Hz3O6N7.C6H307N3: C, 41.4; H, 4.10; (4) On leave of absence from the University, Bristol, England. N, 21.9. Found: C, 41.4; H, 4.17; N, 213). V was treated with HBr-acetic acid and the A SYNTHETIC PREPARATIONPOSSESSJNG BIOLOGI- reaction product isolated as the monohydrobromide CAL PROPERTIES ASSOCIATED WITH ARGININE- (VI) VASOPRESSIN S-Benzyl-N-p-nitrobenzyloxycarbonyl-L-cysSir: teine, m.p. 132.5-133", [LUl2'D -47.0' ( c 1, 95% I n a recent Communication' on the structures of ethanol) (calcd. for ClsH1806N2S: N, 7.18; S, 8.21. arginine-vasopressin and lysine-vasopressin i t was Found: N, 7.05; S, 8.03), as its acid chloride was recorded in a footnote that a synthesis by du Vig- condensed with proline benzyl ester. Saponificaneaud, Popenoe and Roeske of the octapeptide tion of the peptide ester gave S-benzyl-N-p-nitrostructure proposed for lysine-vasopressin had led benzyloxycarbonyl-L-cysteinyl-L-proline(VII) as to biologically active material. As this work con- an oil (neut. equiv., calcd., 487.5. Found, 457). VI was condensed with VI1 by the pyrophosphite tinued, the synthesis of arginine-vasopressin was method' to give S-benzyl-N-p-nitrobenzyloxycarundertaken. hyThe present Communication is concerned with bonyl-L-cysteinyl-L-prolyl-L-arginylglycinamide the preparation and partial purification of a product drobromide (VIII) characterized as the picrate, ~ D (c 1, acetone-water synthesized according to the structure (I) proposed m.p. 182-185", [ c Y ] ~-47.7" C, 47.8; for arginine-vasopressin. The synthesis was (4: 1)) (calcd. for C31H410sNaS.C~H30,N3: approached through the preparation of the appro(3) E. A. Popenoe and V. du Vigneaud, THISJOURNAL, in press. (4) C . R . Harington and R . V. Pitt Rivers, Biochem. J.. 38, 417 priate benzylated cysteine-containing nonapeptide, N-carbobenzoxy-S-benzyl-L- cysteinyl- L - tyrosyl - L- (1944). 'v2
( 5 ) J. R. Vaughan, Jr., and J. A. Eichler, THIS JOURNAL, 76, 5556
(1) V. du Vigneaud, H. 76, 4880 (1983).
C.Lawler and E. A. Popenoe, THISJOURNAL, (1953).
(2) R. Archer and J. Chauvet. Biochim. el Biophys. Acta. (1853).
la,
487
(6) D. T . Gish and F. H. Carpenter, ibid., 76, 5872 (1953). (7) G. W. Anderson, J. Blodinger and A. D. Welcher, ibid., 72, 5309 (1952).
BOOK REVIEW
473%
H, 4.77; K , 18.1. Found: C , 47.8; H, 4.95; N, 17.7). VI11 was treated with HBr-acetic acid and the reaction product isolated as the monohydrobromide (IV). Compounds I11 and IV were coupled by the pyrophosphite method and ether added to the reaction mixture. The resulting precipitate was dried and treated with sodium in liquid ammonia. The product in dilute aqueous solution a t fiH 6.7 was aerated and the solution concentrated and lyophilized. The material from several runs upon assays gave a total of SS,OOO units of pressor activity. After purification by countercurrent distribution, the active material [IS = 0.84 (sa-butyl alcohol-ptoluenesulfonic acid)] was subjected to electrophoresis in a pyridine-acetic acid buffer (pH4) on a cellulose-supporting medium9 The solution from the segment with peak activity assayed 15,500 pressor units and when lyophilized yielded a powder weighing 37 mg. which indicated a specific activity in solution of 400 units/mg. However, the material suffered partial inactivationlo upon lyophilization, assaying 175 units/mg. In countercurrent distribution, electrophoresis and chromatography on partition and ion-exchange (8) J. Dekanski, B r i f . J . Phavinacol., 7 , 567 (1952). (9) H. G . Runkel in “hIethods of Biochemical Analysis,” Vol. I, D. Glick, Ed., Interscience Publishers, Inc., Kern York, N. Y . , p. 141. (10) Samples of highly purified natural vasopressin have also on some occasions shown a loss in activity on concentration a n d lyophilization [R.A. Turner, J. G. Pierce a n d V. d u Vigneaud, J. B i d Chem., 191, 21 (1951); E. A. Popenoe a n d V. d u Vigneaud, J . B i d . Chem., 206, 133 (1953)l. Studies are underway to determine t h e cause as well as t o find means of avoiding such inactivation.
Vol. ‘76
columns, the position of the activity of this final synthetic product was the same as that of the natural arginine-vasopressin. I n addition to the assays against the U.S.P. Standard Powder for pressor activity, the product was assayed for antidiuretic” and avian vasodepressoP activities. The ratios between the pressor, antidiuretic and avian vasodepressor activities were the same as those found for natural vasopressin (1 : 1 :0.15). Further studies on the purification of the synthetic product are underway and i t is hoped that an extensive comparison of its chemical and physical properties with those of natural arginine-vasopressin may eventually be carried out. Since the difficulties are considerable in reaching and maintaining maximum activity, it was felt that a report was warranted at the present time that a synthetic product synthesized according t o the structure proposetl for arginine-vasopressin (I) does possess the expected biological properties. DEPARTMEXT O F BIOCHEMISTRY \TIICE.%?‘ D C ~ I G N E A L ‘ I ) ’ ’ ’ DUASET. G I S H CORNELL UNIV.MEDICALCOLLEGE PANAYOTIS G. KATSOYANSIS’~ NEX YORK,N. Y . RECEIVED ACGUST5, 1954 (11) T h e antidiuretic assays utilizing t h e hydrated normal dog nerc carried o u t by Professor H . B. van Dyke, Dr. IC. Xdamsons, Jr., and Mr. S. L. Engel, t o whom we express our appreciation. (12) J . M . Coon, Arch. i n l e r n . pharmncodynamie, 62, 79 (1939). (13) Appreciation is expressed t o the Lederle Laboratories Division. American Cyanamid Company, for a research grant which has aided greatly in this study. 114) Lilly Postdoctoral Fellow in the Natural Sciences, Kational Research Council (15) Fellow of State Scholarships Foundation of Greece.
BOOK REVIEWS Dictionary of Organic Compounds, 4 Volumes. By SIR biguities which might be anticipated with an alphabetical IANHEILBROX,D.S.O., D S c . , LL.Il., F.R.I.C., F.R.S.,classification have been minimized by the liberal use of cross and H. M. BUNBURY, M.Sc., F.R.I.C. Oxford University references and by carefully stating the nomcriclature rules Press, 114 Fifth Avenue, New York 11, X. Y . 1954, in the introduction. Although Heilbroti’s “Dictionary of Organic C o t n p o u ~ ~ t l s ” T‘olume I, xvi 654 p p . ; Volume IT, xvi 815 p p . ; Yolume 111, xvi 838 pp.; Volume 1ir,xvi 694 p p . ; is not as Comprehensive as Beilstein or Elsevier’s “Ericyclopedia of Organic Chemistry,” and by no means is intended Each Volume--20 X 27 cm. Price, $78.00 a set. to be, i t offers the advantage of being more up to date and During the past twenty years Heilbron’s “Dictionary of convenient to use. Organic Compounds” has become a standard reference work The editors deserve praise for their efforts in making this for chemists concerned with organic compounds. This dicrevised edition available. tionary has proved especially valuable in connection with DEPARTXENr O F CHEMISTRY research and courses involving the characterization of organic I I A K I A N 1,. < > o E R I N G OF WISCONSIN UNIVERSITY compounds. For these reasons i t is fortunate that this MADISON 6, \VISCONSIN \ .iluable reference work has been revised and brought u p t o tlate. O\rer 2500 new entries are included in this revised cclitioti. I n addition, the original entries h a w been brought up to date to the end of 1950 atid in some cases t o 1953. General Biochemistry. By \VILLIAM H. PRTERSON, I>h.lj., After a random check of some of the compounds with which Emeritus Professor of Biochemistry, University of it‘jscorithis reviewer is familiar i t appears t h a t the revision has insin, Madison, and FRASKM. STROSG, Ph.D., Professor (leed been comprehensive. The earlier d a t a have in many of Biochemistry, University of Wisconsin, Madison. instances been replaced or supplemented by more accurate Prentice-Hall, Inc., Englewood Cliffs, New Jersey. 1953. and recent data. v 469 pp. 15.5 X 23.5 cm. Price, $8.65. T h e style and format are essentially the same as in the Modern biochemistry owes its origins to two sources, original edition. The compounds are arranged in alphabetical order and each entry contains the following informa- agriculture and medicine, and has been nourished and sustained by numerous developments in the fundamental tion: structural and molecular formulas, physical propersciences of biology, chemistry and physics. Few scientific t / e s including solubilities, characteristic chemical properareas in recent times include so broad a scope of activity ties, functional derivatives and principal references. Am-
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