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cooling down to room temperature, the mixture was extracted with DCM,
washed with distilled water several times and dried with anhydrous
magnesium sulfate. The crude product was purified by running silica gel
column chromatography using hexane and DCM (v/v, 5/1) as eluting
microtiter plate, followed by measuring by multilabel counter using
430 and 535 nm as excitation and emission wavelengthes, respectively.
The fluorescence of 4 in MOPS/ethanol (8:2, v/v) mixture without bacteria
was also measured for control. The detection limit was calculated by
3 × standard deviation of low concentration/slope of the calibration line.
High-Throughput Antibiotics Screening: Fresh MOPS culturing medium
1
solvent to give 3 as yellow viscous oil (1.76 g, 83%). H NMR (400 MHz,
CDCl , δ): 10.04 (s, −CHO), 10.03 (s, –CHO), 7.95–7.89 (t, 2H), 7.75–
3
6
7
2
.68 (m, 2H), 7.47–7.39 (m, 2H), 7.23–7.09 (m, 12H), 7.05–6.97 (m,
H), 6.73–6.66 (m, 2H), 3.93–3.88 (t, 2H), 1.82–1.73 (m, 2H), 1.52–1.27
was added into EP tube. Then, antibiotics to be screened and 5 × 10
CFU bacteria were introduced to the MOPS culturing medium. The
final volume of each tube was 1 mL. The innoculated bacteria solution
was kept at 37 °C for 4 h. Afterward, 0.25 mL ethanol solution of 4
1
3
(m, 16H), 0.97–0.91 (m, 3H); C NMR (100 MHz, CDCl , δ): 191.176,
3
157.373, 157.278, 146.050, 144.064, 143.993, 143.321, 143.292, 143.233,
140.828, 138.610, 138.572, 136.425, 135.105, 134.446, 131.997, 131.503,
130.873, 130.838, 129.628, 129.609, 127.301, 127.220, 127.178, 127.064,
126.718, 126.010, 125.948, 125.900, 125.861, 113.183, 113.046, 67.240,
31.351, 29.061, 29.019, 28.873, 28.785, 28.745, 25.510, 22.140, 13.609;
−
6
(50 × 10 M) was added to the bacteria solutions, which was then
subjected to incubation at 37 °C for 10 min. Then the solutions were
allocated into 96-well microtiter plate, followed by measuring of
fluorescence intensity employing multilabel counter using 430 and
535 nm as excitation and emission wavelengths, respectively.
HRMS (MALDI-TOF) m/z: calcd for C H O , 606.3498 [M]; found,
4
4
46
2
+
6
06.3506 [M] .
-((1E)-2-(4′-(1,2-Diphenyl-2-(4-(Undecyloxy)Phenyl)vinyl)-
4
[
1,1′-Biphenyl]-4-yl)Vinyl)-1-(3-(Trimethylammonio)Propyl)
Pyridin-1-Ium Bromide (4) : A solution of 1-(3-trimethylammoniopropyl)-4- Supporting Information
methylpyridinium dibromide (0.30 g, 0.85 mmol) and 3 (1 g, 1.63 mmol)
Supporting Information is available from the Wiley Online Library or
from the author.
was refluxed under nitrogen in dry ethanol catalyzed by three drops of
piperidine. After cooling to room temperature, the solvent was removed
by evaporation under reduced pressure. The residue was purified by a
silica gel column chromatography using DCM and methanol mixture
1
(
2:1 v/v) as eluting solvent to give a red powder of 4 (0.38 g, 47%). H Acknowledgements
NMR (400 MHz, DMSO-d , δ): 8.92 (d, 2H), 8.26 (d, 2H), 8.05 (d, 1H),
6
E.Z. and Y.C. contributed equally to this work. This work was in
part supported by the National Basic Research Program of China
7
(
.82–7.71 (m, 4H), 7.59–7.49 (m, 3H), 7.17–6.79 (m, 15H), 6.69–6.61
dd, 2H), 4.59–4.51 (m, 2H), 3.84–3.78 (m, 2H), 3.42–3.35 (m, 3H),
.06 (s, 9H), 2.46–2.36 (m, 2H), 1.64–1.54 (m, 2H), 1.36–1.14 (m, 16H),
(973 Program, 2013CB834701), the University Grants Committee of
3
0
1
1
1
1
3
1
1
3
Hong Kong (AoE/P-03/08), the Research Grants Council of Hong
Kong (604913, 16301614 and N_HKUST604/14), the Innovation and
Technology Commission (ITCPD/17–9), the Science and Technology
Plan of Shenzhen (JCYJ20140425170011516), and the Natural Science
Fund of Guangdong Province (2014A030313659). The authors thank
the support of the Guangdong Innovative Research Team Program
.83–0.76 (m, 3H); C NMR (100 MHz, DMSO-d , δ): 157.045, 153.071,
6
44.138, 143.292, 143.229, 143.160, 140.954, 140.504, 140.465, 138.858,
36.337, 134,971, 133.914, 131.733, 131.273, 130.540, 128.727, 127.792,
27.723, 127.583, 126.714, 126.515, 126.354, 125.784, 125.698, 123.738,
22.915, 113.633, 113.474, 67.055, 61.605, 56.400, 55.883, 52.338,
1.063, 28.727, 28.470, 28.413, 25.259, 23.949, 21.877, 21.866, 18.254,
3.749; HRMS (MALDI-TOF) m/z: calcd for C H Br N O, 861.4353
(201101C0105067115).
5
6
66
2
2
+
+
Note: The labeling of the curves in Figure 4 was incorrect when the
article was initially published: this was corrected on September 2, 2015.
[
M-Br] ; found, 861.4343 [M-Br] .
Sample Preparation:
A
stock solution of
4
in DMSO with a
concentration of 10 × 10 M was prepared and stocked in the 4 °C
fridge. The stock solution was diluted to 50 × 10 M with ethanol prior
to use. MOPS E/Z rich defined medium was prepared according to the
protocol given on the E. coli genome project. Phosphate buffer saline
−
3
−
6
Received: April 24, 2015
Revised: June 18, 2015
Published online: July 14, 2015
(
PBS) was prepared by dissolving NaCl (8 g), KCl (0.2 g), Na HPO4
2
(
1.44 g), and KH PO (0.24 g) in 800 mL distilled water, adjusting pH to
2
4
7
.4 with HCl, and calibrating to 1 L by adding H O. PBS was sterilized
2
−
2
[1] D. Ivnitski, I. Abdel-Hamid, P. Atanasov, E. Wilkins, Biosens. Bioelec-
tron. 1999, 14, 599.
by autoclaving for 20 min at 15 psi (1.05 kg cm ) on liquid cycle and
stored at room temperature.
Bacterial Culturing and Staining: A single colony of bacteria on solid
culture medium [Luria broth (LB) for E. coli and S. epidermidis] was
transferred to 5 mL of liquid culture medium and grown at 37 °C for
[2] L. L. Ling, T. Schneider, A. J. Peoples, A. L. Spoering, I. Engels,
B. P. Conlon, A. Mueller, D. E. Hughes, S. Epstein, M. Jones,
L. Lazarides, V. A. Steadman, D. R. Cohen, C. R. Felix,
K. A. Fetterman, W. P. Millett, A. G. Nitti, A. M. Zullo, C. Chen,
K. Lewis, Nature 2015, 517, 455.
10 h. The concentrations of bacteria were determined by measuring
9
optical density at 600 nm (OD600) and then 10 colony forming unit
9
−1
(
CFU) (1 OD600 = 10 CFU mL ) of bacteria were transferred to a 1.5 mL
[3] J. L. Martínez, F. Baquero, D. I. Andersson, Nat. Rev. Microbiol.
EP tube. Bacteria were harvested by centrifuging at 5000 rpm for 3 min.
2
007, 5, 958.
After removal of supernatant, 1 mL of 4 in PBS solution at the
[
[
[
[
[
[
4] G. Cox, G. D. Wright, Int. J. Med. Microbiol. 2013, 303, 287.
5] F. E. Koehn, G. T. Carter, Nat. Rev. Drug Discovery 2005, 4, 206.
6] J. D. D. Pitout, K. B. Laupland, Lancet Infect. Dis. 2008, 8, 159.
7] G. D. Wright, D. T. Hung, J. D. Helmann, Nat. Med. 2013, 19, 544.
8] M. R. Hamblin, T. Hasan, Photochem. Photobiol. Sci. 2004, 3, 436.
9] A. C. Gales, A. O. Reis, R. N. Jones, J. Clin. Microbiol. 2001, 39, 183.
−6
concentration of 10 × 10 M was added into the EP tube. After dispersing
with vortex, the bacteria were incubated at room temperature for 10 min.
Bacterial Imaging: To take fluorescence images, about 4 µL of stained
bacteria solution was transferred to glass slide and then covered by a
coverslip. The image was collected using 100× objective. The bacteria
were imaged under
00–440 nm excitation filter, 455 nm dichroic mirror, and 465 nm long
pass emission filter.
Standard Curve: Into 1 mL MOPS culture medium was added different
a FL microscope (BX41 Microscope) using
[
10] P. Ertl, E. Robello, F. Battaglini, S. R. Mikkelsen, Anal. Chem. 2000,
2, 4957.
4
7
[
11] R. A. Forsyth, R. J. Haselbeck, K. L. Ohlsen, R. T. Yamamoto, H. Xu,
J. D. Trawick, D. Wall, L. Wang, V. Brown-Driver, J. M. Froelich,
G. C. Kedar, P. King, M. McCarthy, C. Malone, B. Misiner,
D. Robbins, Z. Tan, Z.-Y. Zhu, G. Carr, D. A. Mosca, C. Zamudio,
J. G. Foulkes, J. W. Zyskind, Mol. Microbiol. 2002, 43, 1387.
6
8
−1
concentrations of bacteria (from 5 × 10 up to 2 × 10 CFU mL ), then
−
6
0
.25 mL ethanol solution of 4 (50 × 10 M) was added to the bacteria
solutions. The solutions were then subjected to incubation in 37 °C
incubator for 10 min. Then the solutions were allocated into 96-well
4
936 wileyonlinelibrary.com
© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Adv. Mater. 2015, 27, 4931–4937