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COMMUNICATIONS TO THE EDITOR ULTRAVIOLET IRRADIATION OF l,&DIMETHYLURACIL'
Sir: 1,3-Dirnethyluracil ( I ) z was used to study the interesting but poorly understood ultraviolet irradiation effects on nucleic acids.3 (I) (m.p. 121-122'; : : A 267 mp, E 8.67 X lo3: :;A: 235 rnp, E 1.68 X lo3; A250/A260, A280/A260; 0.62, 0.62 for p H 2-12; vggl 1701 cm-l) in aqueous solution ( O . O G 7 M ) was irradiated with ultraviolet light until SO-SO% of the optical density a t 260 mp had disappeared. The irradiated product (60-75y0 yield) was recrystallized from chloroform and petroleum ether and gave 6-hydroxy-1,3-dimethylhydrouracil (11),4m.p. 105-106' (found: C, 45.61; 1704 H, 6.49; N, 18.00; vggl 3344 cm.-', '::v cm.-l). On treating (11) with acid (pH 2 ) ) alkali (pH 9, 10, 11, 12) or heat, respectively, the ultraviolet spectrum of (I) was reconstituted (A250/ X260, A2SO;A2GO; 0.62, 0.62; 0.61, 0.62; 0.60, 0.62; 0.62, 0.63; 0.61, 0.62). Over 90% yield of ( I ) was obtained in the acidic and alkaline reconstitutions.
Vol. 78
At: 246 mp, e 1.57 X lo3; YE;' 1684 cm.-l). When (I) was treated with excess bromine water, followed by boiling with lead oxide, 5-hydroxy-1,3dimethyluracil (V) m.p. 198-199' (found: C, 46.15; H, 5.38; N, 17.83; A,": 285 r n p , E 7.35 X l o 3 . :A;: 248 mp, e 1.78 X lo3; &' 3215 cm.-l, v?;',"' 1669 cm.-l) was obtained. Hydrogenation of (V) a t two atmospheric pressures gave 5-hydroxy1,3-dimethylhydrouracil (VI), m.p. 109-11 0' (found: C, 45.57; H, 6.40; N, 17.92; ';:v 3390 cm.-', YE:' 1704 cm.-l). No reconstitution of (VI) under similar conditions. When (VI) was irradiated, then treated with acid or alkali, there was no change in ultraviolet spectrum, indicating that (VI) was not converted to (11) on irradiation. These observations suggested strongly that (VI) was not an intermediate. When (I) was treated with one mole equivalent of bromine water trans 5-bromo-6-hydroxy-l,3dimethylhydrouracil (111) was obtained in solution. There was no detectable change in ultraviolet spectrum after standing for two hours. However, when the solution was allowed to stand overnight or crystallized from the CHC13 extract, only (IV) was obtained. This dehydration probably went by way of the "mutarotation" (IIIA) or (IIIB). Hydrogenolysis of (111) in a non-buffered solution gave only (I) as the product. However, hydrogenolysis of (111) in a phosphate buffered solution, pH 7, gave (11), m.p. 105-106' in over 50% yield. This was identical with the irradiated product as determined by m.m.p. 105-106', infrared and analysis (found: C, 45.56; H, 6.49; N, 17.71). (11) was thus identified by synthesis as 6-hydroxy1,3-dimethylhydrouracilI suggesting a 1,4-addition as the mechanism of the photo-reaction. The kinetics of the photo-addition were zero order as shown by a plot of optical density vs. time. The acid catalyzed dehydration (pH 1.98) (IIA), and the base catalyzed @elimination (pH 9.01) (IIB) were both of the first order as shown by a plot of log C"/C, es. time. DEPARTMENT OF PHYSIOLOGY SHIHYI WANG TUFTSUNIVERSITY SCHOOL OF MEDICINE M. APICELLA 136 HARRISON AVENUE B. R. STONE BOSTON11, MASSACHUSETTS RECEIVEDJUNE 22, 1956
sir :
THE BASIC RING SYSTEM OF CRININE
Two recent communications have suggested the likelihood that several alkaloids of the Amaryllidaceue possess structures containing a spiro ring system.'e2 The following results establish a new On treating (I) with an excess of bromine water, spiro ring system for ~ r i n i n e . ~ 5-bromo-1,3-dirnethyluracil (IV), m.p. 184-1 85', Manganese dioxide oxidation of crinine, C15H14was obtained (found: C, 33.03; H, 3.40; N, N(OzCH2) (OH), m.p. 210') gave an a,p-unsatu12.66; Br, 36.20; 283 mp, E 8.57 X lo3; rated ketone, oxocrinine (m.D. 183-185' : found: (1) This work was done under the terms of Contract No. AT(30-1)C, 71.15; H,. 5.52; N, 5.21; "*: : :A 5.98'fi; XEt,"," 911 of the Physiology Department, Tufts University School of Medi227 mfi (4.20). 296 mu (3.59)). Lithium aluminum cine with the Atomic Energy Commission. hydride'reduction o i oiocrinine afforded epicrinine (2) A. M. Moore and C. H. Thomson, Science, 128, 594 (1955). and references therein. (3) E. Chargaff and J. N. Davidson, "The Nucleic Acids," Vol. I, p. 123, Academic Press, Inc., New York, N. Y. (4) The product of Moore and Thomson, m.p. 102'. Following the nomenclature used in Chemical Abstracts (11) should be called 1,3dimethyl-5,6-dihydro-6-hydroxy-2,4-pyri~dinedione.
(1) S. Kobayashi, T. Shingu and S . Uyeo, Chemistry and Induslry 177 (1956). (2) T. Ikeda, W. I. Taylor, Y. Tsuda and S. Uyeo, i b i d . , p. 411. (3) Crinine was reported by L. H. Mason, E. R. Puschett and W. C. Wildman, THISJOURNAL, 77, 1253 (1955); in Chem. Ber., 87, 1704 (1954), H.-G. Boit reported the same alkaloid as "crinidine."
Xug. 20, 1OX
COMMUNICATIONS TO THE
EDITOR
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(m.p. 209-209.5'; found: C, 70.89; H, 6.31; oxocrinine to base and its methohydroxide t o the N, 5.35). Dihydrooxocrinine (m.p. 157-159' ; Hofmann degradation favors structure V. found: C, 70.96; H, 6.17; N, 5.12; al:::, 5.84; Consistent with the morphine-like skeleto~iof "::A: 237 mp (3.56),294 mp (3.72)), obtained by the these alkaloids, several exhibit significant analgesic catalytic hydrogenation of oxocrinine, formed a activity. dibenzylidene derivative (m.p. 125'; found: C, TABLE I 80.31 ; H, 5.88). Under mild WoW-Kishner conPRELIMISARY RESULTS O N ANALGESIC ACTIVITYOF AMMARTI.ditioiis,? dihydrooxocrinine gave 1-crinane (5, lobLIDACEAE ALKALOIDSASD THEIR DERIVAIIVES ethano-8,9-methylenedioxy - 1,2,.7,4,5,6,10b,lOcAlkaloid ED:o LDjo Duration octahydrophenanthridine), (b.p. 130' (1 1); [ o ( ] * ~ D min. -12.7'; found: C, 74.57; H, 7.63; N, 6.37); Buphanidrine 6.2 8.9 106 picrate (m.p. 206-207'; found: C, 54.40; H, dl-Crinane 20 40 ... 4.52; N, 11.51). I-Crinane also was obtained Crinine 45 -100 ,.. from the catalytic hydrogenation-hydrogenolysis Galanthamine 2.7 11.3 141 of crinine. Lycoramine 21.3 89 133 dl-Crinane was synthesized in the following manMorphine 2.1 576 129 ner. 2-(3,4-Methylenedioxyphenyl)-cyclohexa(9) I a m indebted t o Dr. N a t h a n B. E d d y , hTational I n s t i t u t e of none (m.p. 93-94'; found: .C, 71.33; H, 6.55) was obtained from 5-(3,4-methylenedioxyphenyl)-4-ni-Arthritis a n d Metabolic Diseases. for t h e pharmacological data. OF CHEMISTRY OF ~ - A T U R A LPRODUCTS trocy~lohexene~ by the Nef reaction.6 Cyano- LABORATORY ethylation of I and subsequent niethanolysis of XATIOSALHEARTINSTITUTE INSTITUTES OF HEALTH the nitrile gave I1 (m.p. 88-89'; found: C, 67.15; SATIONAL DEPARTMENT OF HEALTH, EDUCATION ASD WELFARE H, 6.51). The action of nitrous acid on the hydra- BETHESDA14, MARYLAND W. C. WILDMAN zone hydrazide' of I1 gave 2,3,4,5,6,7-hexahydroRECEIVED JULY 16, 1966 3a-(3,4-methylenedioxyphenyl)-indolewhich was isolated as the hydroperchlorate (111), (m.p. 227229'; found: C, 52.51; H, 5.27; C1, 10.14). STUDIES ON ADRENOCORTICOTROPIN. XIII. THE ISOLATION OF TWO HIGHLY ACTIVE BOVINE TYPES Catalytic hydrogenation of 111 and Pictet-Spengler cyclization of the resultant base afforded dl- Sir : By means of chromatography on the carboxylic crinane (IV), (m.p. 97-99'; found: C, 74.69; H, 7.27; N, 5.41);picrate ( m p . 218-220'; found: type ion exchange resin XE-9i1 we have separated C, 34.49; H, 4.35; N, 11.GO). The infrared spec- two highly potent types of adrenocorticotropin tra (liquid film) of the I-crinane and the synthetic from beef pituitary extracts. As shown in Fig. 1, crinane were superimposable, as were the spectra the two active types (Al and A*) emerge from the of the natural and synthetic picrates in chloroform column as clearly separated retarded peaks. The distinction between the two peaks in terms of solution. hold-up volumes is maintained when the peptide material recovered from each peak is re-run separately through the same column. The salt-free peptide products recovered3 from the peaks show potencies in the USP test of about 85 units per milligram by the intravenous method. This value is slightly lower than those obtained for the purest porcine4s5and sheep6 preparations, but is several times higher than the best potencies yet obtained for fractions isolated from beef source^.^ The starting material used in our columns was an oxycellulose eluate fraction made from crude ex111 IV tracts of beef anterior glands by a process similar to Crinine must be represented by V or VI. h 3- that described by Astwood, et aL8 As noted by h y d r ~ x y - A ~ -structure ~" is unlikely since dihydro- other^,^ our oxycellulose eluates from beef extracts crinine ( P K a 8.70) is only slightly more basic than have lower potencies than those from pork by a crinine (pKa 7.95).* The stability of dihydroOH
V
VI
(4) 11. Gates a n d G. Tschudi, THIS JOURNAL, 78, 1380 (1956). ( 5 ) 1,. H. Mason a n d W. C. Wildman, ibid., 76, 6194 (1954). (ii! \V. C TVildman and R. R. Wildman. J . Org C h e m . , 17, 581
(l!r;9)
( 7 ) iY E. i%:t.chiiin~iii and I3 J . Pnrnefeld, THISJouan.\~,,73, 31 ( 1 !I.>
I
( 8 ) \.. I'relog aud 0. IIsiiiLrr, IIci?
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d c r j , 31, 1851 (1!149),
(1) Rohm a n d H a a s Co., Washington Square, Philadelphia 5, Pa. ( 2 ) Although we had previously (in our work with pork A C T H ) numbered our chromatographic peaks in t h e order of their emergence from t h e column, we are here adopting t h e reverse convention used b y Dixon and Stack-Dunne in their recent paper (cf. reference 10). (3) For de-salting t h e column fractions, we have used t w o methods with equal success: (a) t h r e e 45 minute dialyses against 10 volumes of 0.1 N acetic acid, o r (b) distribution t o phenol a n d back again t o water. Our procedure i n t h e l a t t e r method is similar t o t h a t of Dixon a n d Stack-Dunne (cf. reference 10) except t h a t we use 0.02 N acetic acid as t h e aqueous phase a n d lyophilize t h e final solution directly. (4) W. F. White, THIS J O U R N A L , 75, 503 (1953). (3) P. H.Bell, i b i d . , 76, 5665 (1964). ( 6 ) C. H. Li, I. I. Geschwind, J. S . Dixon, A. L . Levy a n d J. I. Harris, J. R i d Chrin., 213, 171 (1955). (7) \ V T. Koch a n d F. J . Wolf, THISJ O U R N A L , 77, 489 (I!l>,?). ( 8 ) E. €3. Astwood. R.1. S. Kaben, R . \Y. Payrie and i D (:rads, i b i d . , 73, 2YC,!) (19.51)
