SHORT PAPER
Synthesis and Characterization of Model Ultimate Carcinogens/Metabolites
1461
8 h at 60–65 °C. The reaction mixture was diluted with H2O (50 mL)
and the EtOH was evaporated. The aqueous phase was extracted
successively with Et2O (2 10 mL), butanol (3 10 mL) and EtOAc
(2 10 mL). The combined extracts of Et2O, butanol and EtOAc
were dried (Na2SO4). The crude product obtained on removal of sol-
vent was first purified over a silica gel column with MeOH–CHCl3
(7:2) and then chromatographed on sephadex G-15 with EtOH–
CHCl3 (7:3) to give the product 5a (6 mg, 9%). Analysis of the
aqueous solution by HPLC with H2O–MeCN (6.1) showed that the
product 5a was 99% pure.
Acknowledgement
We are grateful to the Department of Science and Technology
(DST), Government of India, New Delhi, for financial support un-
der project No. SP/S1/GO5/97.
References
(1) (a) Bartsch, H.; Dworkin, M.; Miller, J. A.; Miller, E. C.
Biochim. Biophys. Acta. 1972, 286, 272. (b) Miller, E. C.;
Sandin, R. B.; Miller, J. A.; Rusch, H. P. Cancer Res. 1956,
16, 525. (c) Bouck, N.; Demayorca, G. Nature 1976, 264,
722. (d) Brookers, P.; Lawley, P. D. Nature 1964, 202, 781.
(e) American Chemical Society Monograph, No. 173;
Clayson, D. B.; Garner, R. C.; Searle, C. E., Eds.; ACS:
Washington, 1976, 366. (f) Heidelberger, C. Ann. Rev.
Biochem. 1975, 44, 79. (g) Kadlubar, F. F.; Miller, J. A.;
Miller, E. C. Cancer Res. 1977, 37, 805. (h) Kinosita, R.
Gann. 1936, 30, 423. (i) Berwald, Y.; Sachs, L. Nature
1963, 200, 1182.
1H NMR (400 MHz, DMSO-d6): = 1.82 (m, 1 H, H2 ), 2.14 (s, 3
1
1
1
H, Ar-CH3), 2.60 (m, 1 H, H2 ), 3.61 (m, 2 H, H5 ), 3.82 (d, 1 H,
1
1
1
1
H4 ), 4.36 (m, 1 H, H3 ), 5.28 (s, 1 H, H3 -OH), 5.82 (s, 1 H, H5 -
OH), 6.24 (dd, 1 H, H1 ), 6.38 (s, 2 H, Gu-NH2), 6.92 (d, 2 H, Ar-
1
H), 7.55 (d, 2 H, Ar-H), 7.59–7.82 (m, 5 H, Ar-H), 10.72 (br s, 1 H,
Gu-NH).
IR (KBr): = 3390, 2925, 1684, 1638, 1600, 1516, 1455, 1102,
1050, 1024, 1002, 830 cm–1.
Anal. Calcd for C24H24N6O5: C, 60.50; H, 5.04; N, 17.64. Found: C,
60.50; H, 5.04; N, 17.64.
(2) Padwa, A. Intermolecular 1,3-Dipolar Cycloaddtions, In
Comprehensive Organic Synthesis, Vol. 4; Semmelhack, M.
F., Ed.; Pergamon Press: Oxford, 1991, 1069.
N-Benzoyl-N-(deoxyguanosin-8-yl)aniline (5b)
Compound 4b (173 mg, 0.68 mmol) in 95% EtOH (15 mL) was
added to dG (49 mg, 0.17 mmol) in a 2 mM sodium citrate buffer of
pH 6.9 (30 mL) at 40–45 °C over 2 h and the mixture was stirred
further for 9 h at 60–65 °C. The remaining procedure was the same
as described for the synthesis of 5a to obtain the product 5b (4 mg,
6%). Analysis of the aqueous solution by HPLC with H2O–MeCN
(8:3) showed that the product 5b was 99% pure. The spectral data
were in agreement with that of an authentic sample.14
(3) Butler, R. N.; Scott, F. L.; O'Mahony, A. F. Chem. Rev.
1973, 73, 93.
(4) Hamer, J.; Macaluso, A. Chem. Rev. 1964, 64, 470.
(5) (a) Ohta, A.; Ochai, E. Chem. Pharm. Bull. 1962, 10, 1260.
(b) Ohta, A.; Ochai, E. Chem. Pharm. Bull. 1963, 11, 1586.
(6) (a) Evans, C. A. Aldrichim. Acta. 1979, 12, 23. (b) Janzen,
E. G. Acc. Chem. Res. 1971, 4, 31.
(7) (a) Barton, D. H. R.; Beaton, J. M. J. Am. Chem. Soc. 1960,
82, 2641. (b) Barton, D. H. R.; Beaton, J. M. J. Am. Chem.
Soc. 1961, 83, 4083.
(8) (a) Demeunynck, M.; Lhomme, M.-F.; Mellor, J. M.;
Lhomme, J. J. Org. Chem. 1989, 54, 399. (b) Demeunynck,
M.; Tohme, N.; Lhomme, M.-F.; Mellor, J. M.; Lohme, J. J.
Org. Chem. 1989, 54, 405.
(9) (a) Novak, M.; Rangappa, K. S. J. Org. Chem. 1992, 57,
1285. (b) Novak, M.; Rangappa, K. S.; Manitsas, R. K. J.
Org. Chem. 1993, 58, 7813. (c) Novak, M.; Helmick, J. S.;
Oberlies, N.; Rangappa, K. S.; Clark, W. M.; Swenton, J. S.
J. Org. Chem. 1993, 58, 867.
1
1H NMR (400 MHz, DMSO-d6): = 2.02 (m, 2 H, H2 ), 3.75 (m, 2
1
1
1
H, H5 ), 3.93 (m, 1 H, H4 ), 4.44 (m, 1 H, H3 ), 5.35 (s, 1 H, 1 H,
1
1
1
H3 -OH), 5.96 (s, 1 H, H5 -OH), 6.33 (m, 1 H, H1 ), 6.45 (s, 2 H, Gu-
NH2), 6.91 (t, 1 H, Ar-H), 7.20 (t, 2 H, Ar-H), 7.72–7.82 (m, 7 H,
Ar-H), 10.63 (s,1 H, Gu-NH).
IR (KBr): = 3345, 3206, 1686, 1600, 1564, 1498, 1355, 1098,
1052, 1030, 1000, 752 cm–1.
Anal. Calcd for C23H22N6O5: C, 59.74; H, 4.76; N, 18.18. Found: C,
59.63; H, 4.81; N, 18.22.
(10) (a) Splitter, J. S.; Calvin, M. J. Org. Chem. 1958, 23, 651.
(b) Kamlet, M. J.; Kaplan, L. A. J . Org. Chem. 1957, 22,
576. (c) Shindo, H.; Umezawa, B. Chem. Pharm. Bull. 1962,
10, 492.
(11) Tamagak, S.; Oae, S. Bull. Chem. Soc. Jpn. 1970, 43, 1573.
(12) Galtin, L.; Davis, J. C. J. Am. Chem. Soc. 1962, 84, 1464.
(13) Underwood, G. R.; Price, M. F.; Shapiro, R. Carcinogenesis
1988, 9, 1817.
N-(Benzoyl)-N-(deoxyguanosin-8-yl)-4-Chloroaniline (5c)
Compound 4c (246 mg, 0.85 mmol) in 95% EtOH (15 mL) was add-
ed to dG (49 mg, 0.17 mmol) in a 2 mM sodium citrate buffer of pH
6.9 (30 mL) at 40–45 °C over 2 h and the mixture was stirred further
for 10 h at 60–65 °C. The remaining procedure was the same as de-
scribed for the synthesis of 5a, to give the product 5c (5 mg, 7%).
Analysis of the aqueous solution by HPLC with H2O–MeCN (7:3)
showed that the product 5c was 98.9% pure.
(14) Famulok, M.; Boche, G. Angew. Chem., Int. Ed. Engl. 1989,
28, 468.
1
1H NMR (400 MHz, DMSO-d6): = 2.06 (m, 1 H, H2 ), 2.49 (m, 1
1
1
1
1
H, H2 ), 3.71 (m, 2 H, H5 ), 3.92 (m, 1 H, H4 ), 4.42 (d, 1 H, H3 ),
(15) Meier, C.; Boche, G. Chem. Ber. 1990, 123, 1699.
(16) (a) Meier, C.; Boche, G. Tetrahedron Lett. 1990, 31, 1685.
(b) Meier, C.; Boche, G. Chem. Ber. 1990, 123, 1991.
(c) Meier, C.; Boche, G. Tetrahedron Lett. 1990, 31, 1693.
(17) (a) Patrick, T. B.; Schield, J.; Kirchner, D. G. J. Org. Chem.
1974, 39, 1758. (b) Rondestvedt, C. S.; Johnson, T. A.
Synth. Commun. 1977, 7, 850.
(18) (a) Wheeler, O. H.; Gore, P. H. J. Am. Chem. Soc. 1956, 78,
3363. (b) Beckmann, E. Liebigs Ann. Chem. 1909, 367, 273.
(19) Utzinger, G. E. Liebigs Ann. Chem. 1944, 556, 50.
(20) Kriek, E.; Miller, J. A.; Juhl, U.; Miller, E. C. Biochemistry
1967, 6, 177.
1
1
1
5.37 (s, 1 H, H3 -OH), 6.04 (s, 1 H, H5 -OH), 6.34 (dd, 1 H, H1 ),
6.55 (br s, 2 H, Gu-NH2), 7.28 (d, 2 H, Ar-H), 7.58–7.74 (m, 5 H,
Ar-H), 7.78 (d, 2 H, Ar-H), 10.38 (br s, 1 H, Gu-NH).
IR (KBr): = 3333, 2926, 1682, 1641, 1602, 1495, 1413, 1385,
1092, 1055, 1038, 1012, 826 cm–1.
Anal. Calcd for C23H21ClN6O5: C, 55.59; H, 4.23; N, 16.92. Found:
C, 55.62; H, 4.21; N, 16.99.
Debenzoylation of 5a–c to 6a–c; General Procedure
These compounds were prepared by the method of Underwood et
al.13 using heterogeneous system (Na2CO3/MeOH) for the deben-
zoylation of 5a–c. Spectral data are in good agreement with an au-
thentic sample.14,16b
Synthesis 2001, No. 10, 1459–1461 ISSN 0039-7881 © Thieme Stuttgart · New York