1184
A. S. Hansen et al. / Bioorg. Med. Chem. Lett. 21 (2011) 1181–1184
A.T. gratefully acknowledges Cancer Research UK for a research
studentship in Medicinal Chemistry for Cancer.
Supplementary data
Supplementary data (synthetic protocols and analytical data,
and copies of 1H and 13C NMR spectra for all new compounds. Oli-
gonucleotide synthesis protocol, analysis and characterization of
oligonucleotides) associated with this article can be found, in the
References and notes
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Figure 2. (A) Capillary electrophoresis and (B) electrospray mass spectrum
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Attempts to N-benzoylate 7 with benzoyl chloride following
TMS protection at the 30-hydroxyl group35 afforded complex mix-
tures. However, use of benzoic anhydride in dry DMF25,26 allowed
for the chemoselective protection of the exocyclic amino group of 7
to give 8 (highly acid-sensitive) in 90% isolated yield, again without
reaction at the 30-hydroxyl group.
Compound
8
was then 30-phosphorylated19,20 to afford
the 5-HOMedC phosphoramidite 9 (87% isolated yield, >99% pure by
31P NMR). The 1H NMR of the product was identical to that of a com-
mercial sample (Glen Research Corporation, Sterling, Virginia, USA).
Phosphoramidite 9 was used in the solid-phase synthesis of a series
of oligonucleotides containing up to three 5-HOMedC nucleotides. All
oligonucleotides were analyzed by capillary electrophoresis and
characterized by mass spectrometry (Fig. 2 and Supplementary
data).
In summary, we have developed a substantially improved syn-
thetic route to a protected precursor of 5-hydroxymethylcytosine
and have demonstrated that the product can readily be incorpo-
rated into oligonucleotides for biological applications.
Acknowledgments
This work was supported by the Biotechnology and Biological
Sciences Research Council. A.H.E.-S. and T.B. acknowledge funding
from the European Community’s 7th Framework Programme
[FP7/2007-2013] grant [HEALTH-F4-2008-201418] READNA.
34. Kang, S. B.; De Clercq, E.; Lakshman, M. K. J. Org. Chem. 2007, 72, 5724.
35. Ti, G. S.; Gaffney, B. L.; Jones, R. A. J. Am. Chem. Soc. 1982, 104, 1316.