Supplementary information

Supplementary information
Commercially obtained reagents and solvents were used without further purification. The microwave reaction
was performed in a Smith Synthesizer™ single mode cavity with controlled irradiation at 2450 MHz with a
power of 0-300 W. The reaction temperature was determined using the built-in on-line IR-sensor and the
reaction was executed in a septum sealed process vial. Thin layer chromatography (TLC) was performed using
aluminum sheets precoated with silica gel 60 F254 (0.2 mm, E. Merck). Chromatographic spots were visualized
using UV-detection or by spraying with a 2% ethanolic ninhydrin solution followed by heating, or both. Column
chromatography was performed using commercially available silica gel 60 (particle size: 0.040-0.063 mm).
Analytical RP-HPLC-MS analysis was performed on a Gilson HPLC system with a Finnigan AQA quadropole
mass spectrometer using an Onyx Monolithic C18 4.6 50mm (Phenomenex) and UV detection (DAD) or
ELSD in combination with MS (ESI+) detection, using a MeCN/H2O gradient (0.05% HCOOH). Preparative
HPLC-MS was performed on a Gilson-Finnigan ThermoQuest AQA system equipped with a Sorbax C8 column
(5 m, 21.2 150 mm), using MeCN/H2O (0.05% HCOOH) as the mobile phase with UV (254 nm) and MS
(ESI) detection. The purity the inhibitor was determined by analytical HPLC-MS using the C18 system
described above and Hichrom ACE C4 (5m, 4.6 50mm) with UV detection at 254 nm. 1H and 13C NMR
spectra were recorded on Varian Mercury Plus instruments; 1 H at 399.9 MHz and 13C at 100.6 MHz. Chemical
shifts are reported as δ values (ppm) indirectly referenced to TMS via the solvent signal (1H: CHD2OD δ 3.31,
CHCl3 δ 7.26; 13C: CD3OD δ 49.0, CDCl3 δ 77.16. The exact molecular mass was determined on Micromass QTof2 mass spectrometer equipped with an electrospray ion source.
Synthesis of compound 14.
O
O
N
N
O
a
O
O
O
O
N
H
OH
O
O
S1
H
N
Br
O
N
O
O
H
N
N
H
S2
O
O
b, c
O
S3
O
d
O
N
H
N
H
N
O
Br
H
N
O
O
O
O
H
N
N
H
H
N
S
O O
O
14
Reaction conditions: (a) 2-bromoaniline, HATU, DCM; (b) 4 M HCl/1,4-dioxane; (c) BocTleOH, HATU,
DIPEA, DMF; (d) benzenesulphonamide, Hermann’s palladacycle, [(t-Bu)3PH]BF4, Mo(CO)6, DBU, 1,4dioxane.
(S)-2-(tert-Butoxycarbonylamino)-2-(4-(7-methoxy-2-phenylquinolin-4-yloxy)-phenyl) acetic acid (S1)
Previously described by Örtqvist et al [1].
(S)-tert-Butyl 2-(2-bromophenylamino)-1-(4-(7-methoxy-2-phenylquinolin-4-yloxy)phenyl)-2oxoethylcarbamate (S2)
Compound S2 was synthesized following the method described by Rönn et al [2] using S1 (0.15 g, 0.30 mmol),
2-bromomaniline (0.049 mL, 0.45 mmol), HATU (0.14g, 0.36 mmol) in dry DCM (5 mL). The reaction was run
at 45 °C for 24 h. The organic phase was washed with 0.1 M NaHSO4 (13 mL) and 0.035 M NaHSO4 (23
mL). The organic solvent was evaporated in vacuo, and the crude product was purified by column
chromatography (EtOAc/hexane) to give the title compound as a white solid (0.060 g, 32%). 1H-NMR (CDCl3) 8.34 (dd, J = 8.2, 1.6 Hz, 1H), 8.20-8.16 (m, 2H), 7.96 (m, 2H), 7.59-7.51 (m, 4H), 7.48-7.41 (m, 3H), 7.33 (m,
1H), 7.28 (m, 1H), 7.21 (dd, J = 9.1, 2.6 Hz, 1H), 7.01 (ddd, J = 7.9, 7.5, 1.6 Hz, 1H), 6.98 (s, 1H), 5.90 (bs,
1H), 5.42 (bs, 1H), 4.00 (s, 3H), 1.48 (s, 9H).
13
C-NMR (CDCl3) 168.3,162.0, 161.8, 159.3, 155.4, 152.0,
139.9, 135.2, 134.8, 132.5, 129.7, 129.5, 128.9, 128.7, 127.7, 125.9, 123.0, 122.0, 121.6, 119.2, 115.5, 113.9,
107.7, 101.8, 80.9, 59.6, 55.8, 28.5.
Compound S3
Compound S2 (0.060 g, 0.091 mmol) was dissolved in 4 M HCl/dioxane (3 mL) and stirred at room temperature
for 2 h. The solvent was evaporated and the residue was dried in vacuo. BocTleOH (0.042 g, 0.18 mmol), HATU
(0.083 g, 0.22 mmol) and dry DMF (1.5 mL) was added followed by DIPEA (0.11 mL, 0.62 mmol). The pH of
the solution was controlled and found to be >10. The reaction was stirred at room temperature for 23 h. EtOAc
(20 mL) was added and the organic phase was washed with NaHSO4-buffer (pH 4, 34 mL) and Brine (14
mL). The organic solvent was evaporated in vacuo, and the crude product was purified by column
chromatography (EtOAc/hexane) to give the title compound S3 as a white, solid diastereomeric mixture (0.051
g, 73%). 1H-NMR (CDCl3) 8.21 (dd, J = 8.2, 1.6 Hz, 1H), 8.17-8.11 (m, 5H), 7.95-7.89 (m, 4H), 7.60-7.54
(m, 6H), 7.50-7.45 (m, 2H), 7.44-7.38 (m, 6H), 7.32-7.26 (m, 2H), 7.23-7.16 (m, 4H), 7.02-6.95 (m, 2H), 6.96
(s, 1H), 6.90 (s, 1H), 5.78-5.75 (m, 2H), 5.42 (s, 1H), 5.40 (s, 1H), 4.11 (d, J = 9.6 Hz, 1H), 4.06 (d, J = 9.6 Hz,
1H), 3.98 (m, 6H), 1.40 (s, 9H), 1.37 (s, 9H), 1.05 (s, 9H), 0.95 (s, 9H). 13C-NMR (CDCl3) 171.0, 167.9, 167.8,
162.1, 161.9, 161.8, 159.2, 159.1, 156.0, 155.9, 155.3, 155.1, 151.7, 139.5, 135.1, 135.0, 134.3, 134.0, 132.5,
129.9, 129.7, 129.5, 128.8 (two peaks), 128.5, 127.7, 127.6, 126.3, 126.1, 122.9 (two peaks), 122.5, 121.5,
121.4, 119.2 (two peaks), 115.4, 115.3, 114.2, 107.4, 101.8, 101.6, 79.9, 62.4, 57.8, 55.7, 35.0, 34.9, 28.4, 26.8.
Compound 14
Compound 14 was synthesized as described by Rönn et al [2] using S3 (0.040 g, 0.052 mmol),
benzenesulfonamide (0.041 g, 0.26 mmol), Herrmans palladacycle (0.0024 g, 0.0026 mmol), [(t-Bu)3PH]BF4
(0.0015 g, 0.0052 mmol), Mo(CO)6 (0.014g, 0.052 mmol), DBU (0.023 mL, 0.16 mmol) in 1,4-dioxane (0.3
mL). The reaction was microwave-heated for 15 minutes at 140 °C, after which the solvent was evaporated. The
crude product was dissolved in 15 mL DCM and the organic phase was washed with NaHSO4-buffer (pH 4,
310 mL) and Brine (110 mL). The organic solvent was dried (MgSO4), filtered and evaporated in vacuo. The
crude product was purified by column chromatography (EtOAc/isohexane 1:1 with 5% MeOH) followed by by
RP-HPLC (Sorbax SB-C8, 21.2 150, MeCN/H2O gradient with 0.1% TFA) to give the title compound 14 as a
white solid (0.018 g, 40%) after freeze-drying, 1:1 diastereomeric ratio. 1H-NMR (CD3OD) 8.57 (d, J = 9.2 Hz,
1H), 8.56 (d, J = 9.2 Hz, 1H), 8.09 (dd, J = 8.3, 1.1 Hz, 1H), 8.01-7.98 (m, 5H), 7.77-7.74 (m, 4H), 7.65-7.47
(m, 24H), 7.44-7.41 (m, 4H), 7.23 (m, 1H), 7.22 (m, 1H), 7.01 (s, 1H), 7.00 (s, 1H), 5.61 (s, 1H), 5.56 (s, 1H),
4.10 (s, 6H), 4.02 (s, 1H), 4.01 (s, 1H), 1.42 (s, 9H), 1.41 (s, 9H), 1.02 (s, 9H), 0.96 (s, 9H). 13C-NMR (CD3OD)
173.3, 173.2, 169.7, 169.5, 169.2, 168.9, 168.7, 166.9, 158.4, 158.3, 157.9, 157.8, 154.2, 154.1, 144.4 (two
signals), 141.1, 141.0, 139.0, 138.6, 137.5, 137.3, 134.8 (two signals), 134.5, 134.3, 133.7, 133.2, 132.2, 132.0,
130.7, 130.6, 130.2, 130.1, 130.0 (two signals), 129.7 (two signals), 129.3, 126.1, 125.6, 125.3, 124.2, 123.9,
123.3 (two signals), 122.8, 122.5, 155.8, 103.6 (two signals), 100.8, 80.7, 64.0, 63.8, 59.4, 59.1, 57.0, 35.4 (two
signals), 28.7, 27.2 (two signals). HRMS calcd for C48H50N5O9S (M+H+) 872.3329, found 872.3334. RP-HPLC
purity C18 >99%, C4 99%.
References
1.
Örtqvist P, Peterson SD, Åkerblom E, Gossas T, Sabnis YA, Fransson R, Lindeberg G, Danielson UH,
Karlen A & Sandström A. Phenylglycine as a novel P2 scaffold in hepatitis C virus NS3 protease
inhibitors. Bioorg. Med. Chem. 2007; 15:1448-1474.
2.
Rönn R, Lampa A, Peterson SD, Gossas T, Åkerblom E, Danielson UH, Karlén A & Sandström A.
Hepatitis C virus NS3 protease inhibitors comprising a novel aromatic P1 moiety. Bioorg. Med. Chem.
2008; 16:2955-2967.