4-Acetylamino-1-[2-deoxy-5-O-(4,4ꢀ-dimethoxytrityl)-b-D-erythro-
pentofuranosyl)]-5-(prop-1-ynyl)pyrimidin-2H-2-one
3ꢀ-(2-cyanoethyl)diisopropylphosphoramidite (3b)
Acknowledgements
We thank Mrs E. Michalek, Dr K. I. Shaikh and Dr T. Koch from
Roche Diagnostics GmbH for the measurement of the MALDI
spectra, and Mrs K. Xu and Mr V. R. Sirivolu for reading the
manuscript. Financial support from the Deutsche Forschungsge-
meinschaft, Graduate School 612 and ChemBiotech, Mu¨nster, is
gratefully acknowledged.
Compound 6b (4.1 g, 6.7 mmol) was dissolved in anhyd. CH2Cl2
(10 ml). The mixture was treated with N-ethyldiisopropylamine
(3.1 ml, 17.8 mmol) and (2-cyanoethyl)diisopropylphosphora-
midochloridite (3.1 ml, 14.0 mmol) for 20 min at r.t. The solution
was diluted with CH2Cl2 (10 ml) and poured into 5% aq. NaHCO3
(50 ml). The aqueous layer was extracted with CH2Cl2 (3 × 50 ml),
and the combined organic layers were dried (Na2SO4), filtered
and evaporated. The residual foam was purified by FC (silica
gel, column 10 × 5 cm, CH2Cl2–acetone, 4 : 1). Evaporation of
the main zone afforded a white foam (3b; 3.7 g, 68%). (Found:
C, 65.18; H, 6.39; N, 8.70%. C44H52N5O8P requires C, 65.25; H,
6.47; N, 8.65%); TLC (silica gel, CH2Cl2–acetone, 4 : 1): Rf 0.7. dP
(CDCl3) 149.8, 150.4.
References
1 M. Mills, L. Lacroix, P. B. Arimondo, J.-L. Leroy, J. C. Franc¸ois, H.
Klump and J.-L. Mergny, Curr. Med. Chem.: Anti-Cancer Agents, 2002,
2, 627–644.
2 S. Nonin-Lecomte and J.-L. Leroy, J. Mol. Biol., 2001, 309, 491–506.
3 G. Manzini, N. Yathindra and L. E. Xodo, Nucleic Acids Res., 1994,
22, 4634–4640.
4 J.-L. Leroy, M. Gueron, J.-L. Mergny and C. Helene, Nucleic Acids
´
´ `
Res., 1994, 22, 1600–1606.
5 P. Catasti, X. Chen, L. L. Deaven, R. K. Moyzis, E. M. Bradbury and
G. Gupta, J. Mol. Biol., 1997, 272, 369–382.
6 M. Gue´ron and J.-L. Leroy, Curr. Opin. Struct. Biol., 2000, 10, 326–331.
7 A. T. Phan, M. Gue´ron and J.-L. Leroy, J. Mol. Biol., 2000, 299, 123–
144.
8 A. T. Phan and J.-L. Mergny, Nucleic Acids Res., 2002, 30, 4618–4625.
9 R. D. Wells, D. A. Collier, J. C. Hanvey, M. Shimizu and F. Wohlrab,
FASEB J., 1988, 2, 2939–2949.
10 R. Langridge and A. Rich, Nature, 1963, 198, 725–728.
11 K. Gehring, J.-L. Leroy and M. Gue´ron, Nature, 1993, 363, 561–565.
12 C. H. Kang, I. Berger, C. Lockshin, R. Ratliff, R. Moyzis and A. Rich,
Proc. Natl. Acad. Sci. U. S. A., 1994, 91, 11636–11640.
13 C. H. Kang, I. Berger, C. Lockshin, R. Ratliff, R. Moyzis and A. Rich,
Proc. Natl. Acad. Sci. U. S. A., 1995, 92, 3874–3878.
14 L. Chen, L. Cai, X. Zhang and A. Rich, Biochemistry, 1994, 33, 13540–
13546.
4-[(Dibutylaminomethylidene)amino]-1-[2-deoxy-5-O-(4,4ꢀ-
dimethoxytrityl)-b-D-erythro-pentofuranosyl)]-5-(prop-1-
ynyl)pyrimidin-2H-2-one (6c)
Compound 5 (250 mg, 0.44 mmol) was dissolved in MeOH (5 ml).
Then, N,N-dibutylformamide dimethylacetal (500 ll) was added
and the reaction mixture was stirred for 3 h at r.t. The solvent was
evaporated and the oily residue was co-evaporated with toluene
(3 × 20 ml) and acetone (1 × 10 ml). The resulting oil was purified
by FC (silica gel, column 12 × 4 cm, CH2Cl2–acetone, 9 : 1 →
4 : 1 → 1 : 1) furnishing a colourless foam (6c; 195 mg, 62.7%).
(Found: C, 71.38; H, 7.28; N, 7.78%. C42H50N4O6 requires C, 71.36;
H, 7.13; N, 7.93%); TLC (silica gel, CH2Cl2–acetone, 1 : 1): Rf 0.5;
kmax(MeOH)/nm 340 (e/dm3 mol−1 cm−1 21 200), 257sh (17 800),
236 (38 400); dH (250.13 MHz; [d6]DMSO; Me4Si) 0.92 (6 H, m,
2 × CH3), 1.27 (4 H, m, 2 × CH2), 1.57 (4 H, m, 2 × CH2),
1.79 (3 H, s, CH3), 2.12 (1 H, m, 2ꢀ-H), 2.29 (1 H, m, 2ꢀ-H),
3.15 (2 H, m, NCH2), 3.46 (4 H, m, NCH2 and 5ꢀ-H), 3.74 (6 H,
s, 2 × OCH3), 3.96 (1 H, m, 4ꢀ-H), 4.27 (1 H, m, 3ꢀ-H), 5.33
(1 H, d, J 4.09, 3ꢀ-OH), 6.14 (1 H, ‘t’, J 5.8, 1ꢀ-H), 6.89 (4 H,
m, arom. H), 7.32 (9 H, arom. H), 7.99 (1 H, s, 6-H), 8.58 (1 H,
15 J.-L. Mergny, L. Lacroix, X. Han, J.-L. Leroy and C. He´le`ne, J. Am.
Chem. Soc., 1995, 117, 8887–8898.
16 X. Han, J.-L. Leroy and M. Gue´ron, J. Mol. Biol., 1998, 278, 949–965.
17 D. M. J. Lilley, Biopolymers, 1998, 48, 101–112.
18 M. Edmonds, Bioessays, 1984, 6, 212–216.
19 S. Robidoux, R. Klinck, K. Gehring and M. J. Damha, J. Biomol.
Struct. Dyn., 1997, 15, 517–527.
20 S. Robidoux and M. J. Damha, J. Biomol. Struct. Dyn., 1997, 15, 529–
535.
21 H. Kanehara, M. Mizuguchi, K. Tajima, K. Kanaori and K. Makino,
Biochemistry, 1997, 36, 1790–1797.
22 J.-L. Mergny and L. Lacroix, Nucleic Acids Res., 1998, 26, 4797–4803.
23 J. A. Brazier, J. Fisher and R. Cosstick, Angew. Chem., 2006, 118, 120–
123.
=
s, N CH).
24 L. Lacroix and J.-L. Mergny, Arch. Biochem. Biophys., 2000, 381, 153–
163.
4-[(Dibutylaminomethylidene)amino]-1-[2-deoxy-5-O-(4,4ꢀ-
dimethoxytrityl)-b-D-erythro-pentofuranosyl)]-5-(prop-1-
ynyl)pyrimidin-2H-2-one 3ꢀ-[(2-cyanoethyl)-N,N-
(diisopropyl)]phosphoramidite (3c)
25 F. Seela and Y. He, ‘Modified Nucleosides, Synthesis and Applications’
in: Organic and Bioorganic Chemistry, ed. D. Loakes, Transworld
Research Network, Kerala, India, 2002, pp. 57–85.
26 (a) B. C. Froehler, S. Wadwani, T. J. Terhorst and S. R. Gerrard,
Tetrahedron Lett., 1992, 33, 5307–5310; (b) R. Eritja, E. Ferrer, R.
Gu¨imil and M. Orozco, Nucleosides Nucleotides, 1999, 18, 1619–1621;
(c) V. N. Soyfer and V. N. Potaman, Triple-Helical Nucleic Acids,
Springer-Verlag, New York, 1995, pp. 161–162.
Compound 6c (100 mg, 0.14 mmol) was dissolved in anhyd.
CH2Cl2 (10 ml). The mixture was treated with N,N-diisopropyl-
ethylamine (48 ll, 0.21 mmol) and (2-cyanoethyl)diisopropyl-
phosphoramidochloridite (48 ll, 0.23 mmol) for 20 min at r.t.
The solution was diluted with CH2Cl2 (10 ml) and poured into
5% NaHCO3 solution (50 ml). The aqueous layer was extracted
with CH2Cl2 (3 × 50 ml), and the combined organic layers were
dried (Na2SO4), filtered and evaporated. The residual foam was
purified by FC (silica gel, column 10 × 5 cm, CH2Cl2–acetone,
85 : 15). Evaporation of the main zone afforded a colourless foam
(3c; 81 mg, 63%). TLC (silica gel, CH2Cl2–acetone, 9 : 1): Rf 0.7.
dP (CDCl3) 150.1, 149.5.
´
´
27 J. Sa´gi, A. Szemzo¨, K. Ebinger, A. Szaboles, G. Sa´gi, E Ruff and L.
¨
Otvo¨s, Tetrahedron Lett., 1993, 34, 2191–2194.
28 T. W. Barnes, III and D. H. Turner, Biochemistry, 2001, 40, 12738–
12745.
29 J. He and F. Seela, Nucleic Acids Res., 2002, 30, 5485–5496.
30 F. W. Hobbs, Jr., J. Org. Chem., 1989, 54, 3420–3422.
31 (a) V. Bhat, B. G. Ugarkar, V. A. Sayeed, K. Grimm, N. Kosora, P. A.
Domenico and E. Stocker, Nucleosides Nucleotides, 1989, 8, 179–183;
(b) M. P. Reddy, N. B. Hanna and F. Farooqui, Tetrahedron Lett., 1994,
35, 4311–4314.
32 P. Strazewski, Nucleic Acids Res., 1988, 16, 5191.
33 F. Seela, S. Budow, H. Eickmeier and H. Reuter, Acta Crystallogr.,
Sect. C: Cryst. Struct. Commun., 2007, 63, o54–o57.
This journal is
The Royal Society of Chemistry 2007
Org. Biomol. Chem., 2007, 5, 1858–1872 | 1871
©