F.-S. Liang et al. / Bioorg. Med. Chem. 14 (2006) 1058–1062
1061
layer was washed with brine. The solution was dried
(Na2SO4) and concentrated under reduced pressure.
The product was purified by silica gel column chroma-
tography (EtOAc–hexane = 1:3) to give compound 1
(347 mg, 62%). 1H NMR (500 MHz, CDCl3, 330 K):
d = 7.34–7.31 (m, 2H), 7.29–7.25 (m, 1H), 7.21 (d,
J = 7.4 Hz, 2H), 5.19–5.17 (m, 1H), 4.65 (s, 1H), 3.87–
3.71 (m, 5H), 2.97 (d, J = 5.2 Hz, 1H), 2.94–2.93 (m,
1H), 2.81 (t, J = 4.8 Hz, 1H), 2.76 (s, 1H), 2.12–2.08
(m, 1H), 1.98–1.94 (m, 1H), 1.63 (s, 1H); HRMS (ESI)
m/z calcd for C15H20NO4 [M+H]+: 278.1387. Found:
278.1392.
14.7 Hz, 4H), 5.09 (s, 1H), 5.02 (d, J = 8.8 Hz, 1H),
3.86 (s, 3H), 3.82–3.72 (m, 4H), 3.61–3.59 (m, 2H),
3.53 (d, J = 4.8 Hz, 1H), 3.00–2.97 (m, 1H), 2.92–2.87
(m, 1H), 2.9–2.04 (m, 1H), 1.91–1.89 (m, 1H); HRMS
(ESI) m/z calcd for C34H37N2O8S [M+H]+: 633.2265.
Found: 633.2262.
1
Compound 7: H NMR (500 MHz, CDCl3, 330 K): d
7.70 (d, J = 8.8 Hz, 2H), 7.30–7.27 (m, 2H), 7.23–7.19
(m, 3H), 6.97 (d, J = 8.8 Hz, 2H), 5.10–5.07 (m, 2H),
3.87–3.81 (m, 7H), 3.77–3.74 (m, 2H), 3.61–3.58 (m,
1H), 3.14–3.09 (m, 1H), 3.04–3.00 (m, 2H), 2.96–2.92
(m, 1H), 2.88–2.79 (m, 2H), 2.10–2.04 (m, 1H), 1.92–
1.89 (m, 1H), 1.86–1.81 (m, 1H), 0.90 (d, J = 6.6 Hz,
3H), 0.86 (d, J = 7.0 Hz, 3H); HRMS (ESI) m/z calcd
for C26H37N2O7S [M+H]+: 521.2316. Found: 521.2317.
4.2. General procedure for library synthesis and screening
An 18 mM solution of epoxide core was prepared in
DMSO–H2O (1:1). 100 lL of this solution was dis-
pensed into each well of a 96-well microtiter plate, which
also contained the corresponding amine (8 equiv). The
reactions were allowed to stand in a 60 °C oven 6 h
and analyzed by LC–MS. The screening of synthesized
inhibitors was done following reported procedures.20
AB2,22 a reported potent inhibitor, was used as a posi-
tive control and DMSO was used as a negative control.
Acknowledgments
We thank the National Institutes of Health and The
Skaggs Institute for Chemical Biology for financial
support.
References and notes
4.3. General procedure for inhibitor synthesis
1. (a) Brik, A.; Lin, Y.-C.; Elder, J.; Wong, C.-H. Chem.
Biol. 2002, 9, 891; (b) Brik, A.; Muldoon, J.; Lin, Y.-C.;
Elder, J.; Goodsell, D. S.; Olson, A. J.; Fokin, V. V.;
Sharpless, K. B.; Wong, C.-H. ChemBioChem 2003, 4,
1246; (c) Cheng, T.-J.; Brik, A.; Wong, C.-H.; Kan, C.-C.
Antimicrob. Agents Chemother. 2004, 48, 2437(d) Wu, C.-
Y.; Brik, A.; Wang, S.-K.; Chen, Y.-H.; Wong, C.-H.
ChemBioChem, in press.
2. Wu, C.-Y.; Jan, J.-T.; Ma, S.-H.; Kuo, C.-J.; Juan, H.-F.;
Cheng, Y.-S. E.; Hsu, H.-H.; Huang, H.-C.; Wu, D.; Brik,
A.; Liang, F.-S.; Liu, R.-S.; Fang, J.-M.; Chen, S.-T.;
Liang, P.-H.; Wong, C.-H. Proc. Natl. Acad. Sci. U.S.A.
2004, 101, 10012.
3. (a) Wu, C.-Y.; Chang, C.-F.; Chen, S.-Y.; Wong, C.-H.;
Lin, C.-H. Angew. Chem. 2003, 115, 4809; . Angew. Chem.,
Int. Ed. 2003, 42, 4661; (b) Chang, C.-F.; Ho, C.-W.; Wu,
C.-Y.; Chao, T.-A.; Wong, C.-H.; Lin, C.-H. Chem. Biol.
2004, 11, 1301.
4. (a) Best, M. D.; Brik, A.; Chapman, E.; Lee, L. V.; Cheng,
W.-C.; Wong, C.-H. ChemBioChem 2004, 5, 811; (b) Brik,
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A 100 mM solution of tert-butyl [S-(R*,R*)]-(ꢀ)-(1-
oxiranyl-2-phenylethyl)carbamate (2) (1 equiv) in 2-pro-
panol was treated with an amine (6 equiv) and stirred at
60 °C overnight. The reaction mixture was then concen-
trated in vacuo and purified by silica gel column chro-
matography. The resulting product was then dissolved
in dry CH3CN (100 mM) and were added dry pyridine
(1.5 equiv) and p-methoxybenzenesulfonyl chloride
(1 equiv) under argon. The reaction mixture was stirred
at room temperature overnight, then concentrated in
vacuo, and purified by silica gel column chromatogra-
phy. The product was treated with TFA–CH2Cl2 (1:1)
and stirred at room temperature. After 1 h, toluene
was added and the reaction was concentrated in vacuo.
The crude product was then treated with freshly pre-
pared (S)-3-tetrahydrofuranyl N-oxysuccinimidyl car-
bonate (2 equiv) and dry triethylamine (5 equiv) in dry
CH2Cl2. The reaction was stirred at room temperature
overnight under argon, and then concentrated in vacuo
and purified by silica gel column chromatography to
give the pure inhibitor.
5. Lee, L. V.; Mitchell, M. L.; Huang, S.-J.; Fokin, V. V.;
Sharpless, K. B.; Wong, C.-H. J. Am. Chem. Soc. 2003,
125, 9888.
1
Compound 5: H NMR (500 MHz, CDCl3, 330 K): d
6. Bonini, C.; Righi, G. Synthesis 1994, 225.
7.58 (d, J = 8.5 Hz, 2H), 7.45 (s, 1H), 7.28–7.24 (m,
2H), 7.22–7.19 (m, 1H), 7.16–7.15 (m, 2H), 7.00 (d,
J = 8.1 Hz, 1H), 6.93 (d, J = 8.5 Hz, 2H), 6.88 (d,
J = 8.4 Hz, 1H), 5.11–5.06 (m, 2H), 3.91–3.81 (m, 6H),
3.76–3.73 (m, 2H), 3.61 (d, J = 10.7 Hz, 1H), 2.96–2.92
(m, 1H), 2.82–2.78 (m, 1H), 2.11 (s, 3H), 2.11–2.04 (m,
1H), 1.95–1.92 (m, 2H); HRMS (ESI) m/z calcd for
C29H35N2O8S [M+H]+: 571.2109. Found: 571.2107.
7. In 1995, thirteen of the top two hundred drugs ranked by
prescription volume were ethanolamine-based compounds
(source: Pharmacy Times, April 1996).
8. Brik, A.; Wong, C.-H. Org. Biomol. Chem. 2003, 1, 5.
9. Kim, E. E.; Baker, C. T.; Dwyer, M. D.; Murcko, M. A.;
Rao, B. G.; Tung, R. D.; Navia, M. A. J. Am. Chem. Soc.
1995, 117, 1181.
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11. Roberts, N. A.; Martin, J. A.; Kinchington, D.; Broad-
hurst, A. V.; Craig, J. C.; Duncan, I. B.; Galpin, S. A.;
Handa, B. K.; Kay, J.; Krohn, A.; Lambert, R. W.;
Merrett, J. H.; Mills, J. S.; Parkes, K. E. B.; Redshaw, S.;
1
Compound 6: H NMR (500 MHz, CDCl3, 330 K): d
7.51 (d, J = 8.8 Hz, 2H), 7.36 (t, J = 8.1 Hz, 2H), 7.30–
7.26 (m, 2H), 7.23–7.14 (m, 4H), 7.03 (d, J = 8.1 Hz,
2H), 6.97 (d, J = 8.8 Hz, 2H), 6.91 (dd, J = 8.8,