www.advancedsciencenews.com
www.afm-journal.de
1
Synthesis of TPE-2M2F N-Oxide: Yield: 85%. H NMR (CDCl , 400 MHz),
A-HKUST 605/16, C6009-17G, and N_HKUST604/14), the Innovation
and Technology Commission (ITC-CNERC14SC01, ITCPD/17-9, and
ITCPD/17-9), and the Science and Technology Plan of Shenzhen
(JCYJ20170818113602462 and JCYJ20160229205601482).
3
δ (TMS, ppm):7.71–7.73 (d, 4H, J = 8.8 Hz), 7.06–7.12 (d, 4H, J = 8.8 Hz),
6
1
1
.92–7.00 (m, 4H), 6.78–6.86 (m, 4H), 3.56 (s, 12H). 13C NMR (CD OD,
3
00 MHz), δ (TMS, ppm):163.1, 160.7, 152.0, 144.1, 141.8, 138.6, 138.6,
38.3, 132.7, 132.6, 131.8, 119.5, 114.7, 114.5, 61.6. HRMS (MALDI-TOF),
m/z calcd. for C H F N O : 486.2119; found 454.2207 [C H F N ] .
+
30
28
2
2
2
30 28
2
2
Cell Culture: HeLa cells were cultured in minimum Eagle’s medium
−
1
(
MEM)containing10%fetalbovineserum(Invitrogen)and100unitsmL
Conflict of Interest
−
1
penicillin and 100 mg mL streptomycin. COS-7 cells were cultured
in Dulbecco’s modified Eagle medium supplemented with 10%
fetal bovine serum (Invitrogen) and 100 units mL penicillin and
The authors declare no conflict of interest.
−
1
−
1
1
00 mg mL streptomycin. The cells were incubated in a standard
incubator, maintained at 37 °C under 5% CO and 21% O , providing
2
2
Keywords
a normoxic condition. Hypoxic incubator was performed by incubating
the cells in a 2.5 L rectangular sealed container with Anaero Pack-Anaero
anaerobic gas generator (0.1% O2 concentration) or AnaeroPack-
MicroAero gas generator (8% O2 concentration). Hypoxic conditions
were confirmed by the use of MGC RT Anaero-Indicator.
aggregation-induced emission, hypoxia, N-oxide, sensitivity, water
solubility
Received: April 24, 2019
Revised: May 30, 2019
Published online:
Cell Imaging: Cells were cultured overnight in a 35 mm Petri dish
with coverslip. The cell medium was replaced with fresh one and
−
1
the cells were stained with 100 μg mL of TPE-2M N-oxide, TPE-2E
N-oxide, or TPE-2M2F N-oxide for 3 h under different oxygen conditions.
Subsequently, the cell coverslip was taken out and the dish was mounted.
The cells were imaged using a laser scanning confocal microscope
[
1] J. Bernier, Head and Neck Cancer: Multimodality Management,
Springer Science & Business Media, Berlin 2011.
(LSM710, Zeiss) at 405 nm with 2% laser power. The emission filter was
4
30–560 nm.
Flow Cytometry: Cells were seeded and treated with TPE-2E N-oxide
[2] P. Vaupel, L. Harrison, Oncologist 2004, 9, 4.
under different oxygen conditions for 3 h. The fluorescence was
measured using a Becton–Dickinson flow cytometer Aria IIIu with an
excitation wavelength of 405 nm. 10 000 cells were counted. The data
were analyzed using Flowjo.
[3] Y. Cao, C.-Y. Li, B. J. Moeller, D. Yu, Y. Zhao, M. R. Dreher, S. Shan,
M. W. Dewhirst, Cancer Res. 2005, 65, 5498.
[4] a) M. Zhang, Z. Zhou, Z. Zhao, J. Garrison, Explor. Res. Hypoth-
esis Med. 2016, 1, 17; b) W.-J. Koh, K. S. Bergman, J. S. Rasey,
L. M. Peterson, M. L. Evans, M. M. Graham, J. R. Grierson,
K. L. Lindsley, T. K. Lewellen, K. A. Krohn, Int. J. Radiat. Oncol., Biol.,
Phys 1995, 33, 391.
Cytotoxicity Study: Cell viability was evaluated by MTT assay. Cells were
seeded in 96-well plates overnight at a density of 5000–8000 cells per
well. Then the medium was replaced with 200 μL fresh MEM medium
containing various concentrations of TPE-2M N-oxide, TPE-2E N-oxide, or
TPE-2M2F N-oxide, and the cells were incubated under normoxic or hypoxic
conditions. After 24 h, 10% MTT solution was added into the medium,
followed by adding 100 μL dimethyl sulfoxide. The plates were analyzed with
a microplate reader (Varioskan LUX multimode microplate reader) with the
absorbance at 595 nm. Each experiment was performed at least three times.
Inhibitor Assay: A certain amount of DPI was dissolved in Milli-Q
water to serve as a stock solution. The cells were incubated with TPE-2M
N-oxide, TPE-2E N-oxide, or TPE-2M2F N-oxide in the presence of
[
5] M. W. Gross, U. Karbach, K. Groebe, A. J. Franko, W. Mueller-Klieser,
Int. J. Cancer 1995, 61, 567.
[
6] a) W. Piao, S. Tsuda, Y. Tanaka, S. Maeda, F. Liu, S. Takahashi,
Y. Kushida, T. Komatsu, T. Ueno, T. Terai, Angew. Chem., Int. Ed.
2
013, 52, 13028; b) H. Komatsu, H. Harada, K. Tanabe, M. Hiraoka,
S.-i. Nishimoto, MedChemComm 2010, 1, 50; c) S. Luo, R. Zou,
J. Wu, M. P. Landry, ACS Sens. 2017, 2, 1139; d) P. Zhang,
H. Huang, Y. Chen, J. Wang, L. Ji, H. Chao, Biomaterials 2015, 53,
522.
−
6
−6
various concentrations of DPI (0, 300 × 10 , and 500 × 10 m) under
different oxygen conditions for 3 h.
[
7] a) K. Kiyose, K. Hanaoka, D. Oushiki, T. Nakamura, M. Kajimura,
M. Suematsu, H. Nishimatsu, T. Yamane, T. Terai, Y. Hirata, J.
Am. Chem. Soc. 2010, 132, 15846; b) S. Luo, Y. Liu, F. Wang,
Q. Fei, B. Shi, J. An, C. Zhao, C.-H. Tung, Analyst 2016, 141,
2879; c) L. Cui, Y. Shi, S. Zhang, L. Yan, H. Zhang, Z. Tian,
Y. Gu, T. Guo, J. Huang, Dyes Pigm. 2017, 139, 587; d) L. Sun,
G. Li, X. Chen, Y. Chen, C. Jin, L. Ji, H. Chao, Sci. Rep. 2015, 5,
Confocal Colocalization: HeLa cells were incubated under hypoxic
condition with TPE-2E N-oxide for 3 h. The medium was removed and
washed with phosphate buffered saline buffer solution. Then Nile red
−1
(
100 ng mL ) was added for 15 min. Imaging of the HeLa cells was
conducted under a confocal microscope. The excitation wavelength
of the reduced form of TPE-2E N-oxide was 405 nm, and the emission
filter was 430–560 nm. For Nile red, the excitation was 560 nm and the
emission filter was 580–700 nm.
1
4837.
[
8] a) J. Luo, Z. Xie, J. W. Lam, L. Cheng, H. Chen, C. Qiu, H. S. Kwok,
X. Zhan, Y. Liu, D. Zhu, B. Z. Tang, Chem. Commun. 2001, 1740;
b) Y. Hong, J. W. Lam, B. Z. Tang, Chem. Soc. Rev. 2011, 40, 5361;
c) J. Mei, N. L. Leung, R. T. Kwok, J. W. Lam, B. Z. Tang, Chem.
Rev. 2015, 115, 11718; d) J.-S. Ni, H. Liu, J. Liu, M. Jiang, Z. Zhao,
Y. Chen, R. T. Kwok, J. W. Y. Lam, Q. Peng, B. Z. Tang, Mater.
Chem. Front. 2018, 2, 1498; e) Y. Zhang, W. Xu, L. Kong, B. Han,
Z. Cai, J. Shi, B. Tong, Y. Dong, B. Z. Tang, Mater. Chem. Front.
Supporting Information
Supporting Information is available from the Wiley Online Library or
from the author.
2
018, 2, 1779; f) V. G. Naik, S. D. Hiremath, A. Das, D. Banwari,
R. U. Gawas, M. Biswas, M. Banerjee, A. Chatterjee, Mater. Chem.
Front. 2018, 2, 2091.
Acknowledgements
[
9] a) N. L. Leung, N. Xie, W. Yuan, Y. Liu, Q. Wu, Q. Peng, Q. Miao,
J. W. Lam, B. Z. Tang, Chem. - Eur. J. 2014, 20, 15349; b) F. Bu,
R. Duan, Y. Xie, Y. Yi, Q. Peng, R. Hu, A. Qin, Z. Zhao, B. Z. Tang,
Angew. Chem., Int. Ed. 2015, 54, 14492.
C.X. and H.Z. contributed equally to this work. This work was partially
supported by the National Natural Science Foundation of China (Grant
No. 21788102), the University Grants Committee of Hong Kong
(AoE/P-03/08), the Research Grants Council of Hong Kong (16305015,
Adv. Funct. Mater. 2019, 1903278
1903278 (11 of 12)
© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim