Organic & Biomolecular Chemistry
Paper
syringe. Throughout the reaction a constant stream of argon HRMS (EI): C7H2OND2 125.0804; found 125.0803. 2,6-Dideu-
was purged through the reaction mixture via a syringe and terio-p-anisidine 29 (96 mg) was treated with HBr (47%,
needle to prevent oxidative degradation of the product. The 620 mL) and Aliquat-336® (16 mg) at 105 °C for 6 h. The reac-
mixture was stirred at 110 °C for 4 h. After the reaction was tion was quenched by the addition of water (5 mL) and the
allowed to cool to room temperature, the mixture was extracted resulting solution was extracted with ethyl acetate (50 mL).
with diethylether (2 × 20 mL). Subsequent addition of HClconc. After the aqueous phase was brought to a pH > 12 by addition
(2.5 mL) to the aqueous phase was followed by extraction with of 1 M NaOH, the mixture was extracted with ethyl acetate
diethylether (5 × 30 mL). The organic phases were combined (3 × 50 mL). The organic phases resulting from the second
and dried over MgSO4. Evaporation of the solvents under extraction were combined, washed with water (2 × 20 mL) and
reduced pressure gave a white solid to which D2O was added dried over MgSO4. The crude product was purified over a short
(10 mL) and the resulting solution was brought to pH 7 by silica-gel column using ethyl acetate as an eluent to yield
slow addition of NaOD (1 N). Lyophilization yielded sodium 3,5-dideuterio-4-aminophenol 30 (63 mg, 73%). 1H NMR
3,3-dideuterio([3,5-13C2]2,6-dideuterio-4-hydroxyphenyl)pyruvate (400 MHz, DMSO-d6): 8.30 (s, 1H, OH), 6.46 (s, 2H, o-CHarom.),
3 (44 mg, 88%) as a yellow solid. NMR analysis showed 4.34 (s, 2H, NH2); 13C NMR (100.6 Hz, DMSO-d6): 148.16
1
residual H at C3 (<4%). 1H NMR (400 MHz, D2O): 6.83 (dm, (Carom.OH), 140.52 (Carom.NH2), 115.07 (o-CHarom.).
J = 163.9 Hz, 1.5 H, 13CHarom.), 3.99 (s, 0.07 H, residual
CHarom.); 13C NMR (100.6 Hz, D2O): 115.69 (13C). HRMS
(ESI): calcd for C713C2H3D4O4 [M − Na]− 185.0663; found
185.0663.
Notes and references
3,5-Dideuterio-4-aminophenol 30. 1,8-Diazabicyclo[5.4.0]-
undec-7-ene (DBU, 750 µL) was added to a stirred solution of
4-nitrophenol (700 mg) in acetone (25 mL). The reaction
mixture was stirred at room temperature for 10 min before
iodomethane (934 µL) was added dropwise. After stirring the
reaction mixture for 4 h, TLC still showed remaining starting
material. Therefore, additional DBU (750 µL) and iodomethane
(310 µL) were added and stirring was continued for 1 h. The
solvents were then removed under reduced pressure and the
residue was dissolved in ethyl acetate (100 mL). This solution
was washed with 1 N HCl (10 mL), water (10 mL), a saturated
solution of sodium thiosulfate (10 mL) and brine (10 mL).
Drying of the organic phase over MgSO4 and evaporation of
1 U. Weininger, C. Diehl and M. Akke, J. Biomol. NMR, 2012,
53, 181–190.
2 L. Lo Conte, C. Chothia and J. Janin, J. Mol. Biol., 1999,
285, 2177–2198.
3 (a) Z. Lin, Y. Xu, S. Yang and D. Yang, Angew. Chem., Int.
Ed., 2006, 45, 1969–1963, (Angew. Chem., 118, 1994–1997);
(b) C. M. Slupsky, L. N. Gentile and L. P. McIntosh,
Biochem. Cell Biol., 1998, 76, 379–390; (c) G. W. Vuister,
S.-J. Kim, C. Wu and A. Bax, J. Am. Chem. Soc., 1994, 116,
9206–9210.
4 J. A. Boyer and A. L. Lee, Biochemistry, 2008, 47(17), 4876–
4886.
5 (a) M. Takeda, A. M. Ono, T. Terauchi and M. Kainosho,
J. Biomol. NMR, 2010, 46, 45–49; (b) P. Lundström,
K. Teilum, T. Carstensen, I. Bezsonova, S. Wiesner,
D. Flemming Hansen, T. L. Religa, M. Akke and L. E. Kay,
J. Biomol. NMR, 2007, 38, 199–212.
6 (a) V. Kasinath, K. G. Valentine and A. J. Wand, J. Am.
Chem. Soc., 2013, 135, 9560–9563; (b) K. Teilum, U. Brath,
P. Lundström and M. Akke, J. Am. Chem. Soc., 2006, 128,
2506–2507.
7 M. Kainosho, T. Torizawa, Y. Iwashita, T. Terauchi, A. Mei
Ono and P. Güntert, Nature, 2006, 440, 52–57.
8 S. Rajesh, D. Nietlispach, H. Nakayama, K. Takio,
E. D. Laue, T. Shibata and Y. Ito, J. Biomol. NMR, 2003, 27,
81–86.
1
the solvents yielded 4-nitroanisole 27 (686 mg, 90%). H NMR
(400 MHz, CDCl3): 8.21 (dm, J = 9.3 Hz, 2H, m-CHarom.), 6.96
(dm, J = 9.3 Hz, 2H, o-CHarom.), 3.91 (s, 3H, CH3); 13C NMR
(100.6 Hz, CDCl3): 164.59 (Carom.), 125.92 (CHarom.), 114.02
(CHarom.), 55.95 (CH3). 4-Nitroanisole 27 (546 mg) was dis-
solved in MeOH (40 mL) and conducted over a Pd/C (10%)
catalyst cartridge in
a continuous-flow hydrogen reactor
(H-cube® – Thalesnano) at a flow-rate of 1 mL min−1 and
room temperature. The hydrogen generator was set to full
hydrogen mode. Evaporation of methanol under reduced
pressure gave p-anisidine 28 (392 mg, 89%). 1H NMR
(400 MHz, DMSO-d6): 6.74 (dm, J = 8.9 Hz, 2H, m-CHarom.),
6.65 (dm, J = 8.9 Hz, 2H, o-CHarom.), 3.75 (s, 3H, CH3), 4.41 (bs,
2H, NH2); 13C NMR (100.6 Hz, DMSO-d6): 152.87 (Carom.),
139.94 (Carom.NH2), 116.42 (CHarom.), 114.85 (CHarom.), 55.76
9 C. G. Hoogstraten and J. E. Johnson Jr., Concepts. Magn.
Reson. A, 2008, 32A, 34–55.
(CH3). A microwave vessel was charged with anisidine 28 10 R. J. Lichtenecker, K. Weinhäupl, W. Schmid and
(272 mg), D2O (2.5 mL) and HClconc. (50 µL) and heated in the R. Konrat, J. Biomol. NMR, 2013, 57(4), 327–331.
microwave reactor at 180 °C for 40 min. After the solvents 11 (a) R. Lichtenecker, M. L. Ludwiczek, W. Schmid and
had been removed under reduced pressure, the residue was
dissolved in methanol (10 mL) and concentrated again to yield
2,6-dideuterio-p-anisidine 29 (245 mg, 90%). 1H NMR
(400 MHz, DMSO-d6): 6.73 (s, 2H, CHarom.), 6.37 (bs, 2H, NH2),
3.65 (s, 3H, CH3); 13C NMR (100.6 Hz, DMSO-d6): 153.14
(Carom.), 138.15 (Carom.NH2), 114.95 (CHarom.), 55.81 (CH3).
R. Konrat, J. Am. Chem. Soc., 2004, 126(17), 5348–5349;
(b) M. Fischer, K. Kloiber, J. Hauesler, K. Ledolter,
R. Konrat and W. Schmid, ChemBioChem, 2007, 8(6), 610–
612; (c) R. J. Lichtenecker, N. Coudevylle, R. Konrat and
W. Schmid, ChemBioChem, 2013, 14(7), 818–821;
(d) R. J. Lichtenecker, K. Weinhäupl, L. Reuther,
This journal is © The Royal Society of Chemistry 2014
Org. Biomol. Chem., 2014, 12, 7551–7560 | 7559