A simple nitrobenzyl-umbelliferone (NCOU1) was synthesised containing a nitroreductase (NTR) trigger moiety. The presence of NTR, resulted in the fragmentation of the parent molecule and release of the highly emissive fluorophore umbelliferone via an NTR-catalyzed reduction of the nitro group. In the presence of the NTR enzyme, NCOU1 gave rise to a 5-fold increase in fluorescence intensity at 455 nm and was selective for NTR over other reductive enzymes. These results indicate that NCOU1 can be used as a simple assay for the detection of NTR.
收稿日期: 2017-09-17
出版日期: 2018-05-09
Corresponding Author(s):
Robert B. P. Elmes,Tony D. James
引用本文:
. [J]. Frontiers of Chemical Science and Engineering, 2018, 12(2): 311-314.
Adam C. Sedgwick, Alex Hayden, Barry Hill, Steven D. Bull, Robert B. P. Elmes, Tony D. James. A simple umbelliferone based fluorescent probe for the detection of nitroreductase. Front. Chem. Sci. Eng., 2018, 12(2): 311-314.
Elmes R B P. Bioreductive fluorescent imaging agents: Applications to tumour hypoxia. Chemical Communications, 2016, 52(58): 8935–8956 https://doi.org/10.1039/C6CC01037G
5
Pacheco-Torres J, López-Larrubia P, Ballesteros P, Cerdán S. Imaging tumor hypoxia by magnetic resonance methods. NMR in Biomedicine, 2011, 24(1): 1–16 https://doi.org/10.1002/nbm.1558
6
Wu J, Kwon B, Liu W, Anslyn E V, Wang P, Kim J S. Chromogenic/fluorogenic ensemble chemosensing systems. Chemical Reviews, 2015, 115(15): 7893–7943 https://doi.org/10.1021/cr500553d
7
Yang Z, Cao J, He Y, Yang J H, Kim T, Peng X, Kim J S. Macro-/micro-environment-sensitive chemosensing and biological imaging. Chemical Society Reviews, 2014, 43(13): 4563–4601 https://doi.org/10.1039/C4CS00051J
8
Qian X, Xiao Y, Xu Y, Guo X, Qian J, Zhu W. “Alive” dyes as fluorescent sensors: Fluorophore, mechanism, receptor and images in living cells. Chemical Communications, 2010, 46(35): 6418–6436 https://doi.org/10.1039/c0cc00686f
9
Xu K, Wang F, Pan X, Liu R, Ma J, Kong F, Tang B. High selectivity imaging of nitroreductase using a near-infrared fluorescence probe in hypoxic tumor. Chemical Communications, 2013, 49(25): 2554–2556 https://doi.org/10.1039/c3cc38980d
10
Wan Q Q, Gao X H, He X Y, Chen S M, Song Y C, Gong Q Y, Li X H, Ma H M. A cresyl violet-based fluorescent off-on probe for the detection and Imaging of hypoxia and nitroreductase in living organisms. Chemistry, an Asian Journal, 2014, 9(8): 2058–2062 https://doi.org/10.1002/asia.201402364
11
Yuan J, Xu Y Q, Zhou N N, Wang R, Qian X H, Xu Y F. A highly selective turn-on fluorescent probe based on semi-cyanine for the detection of nitroreductase and hypoxic tumor cell imaging. RSC Advances, 2014, 4(99): 56207–56210 https://doi.org/10.1039/C4RA10044A
12
Wong R H F, Kwong T, Yau K H, Au-Yeung H Y. Real time detection of live microbes using a highly sensitive bioluminescent nitroreductase probe. Chemical Communications, 2015, 51(21): 4440–4442 https://doi.org/10.1039/C4CC10345A
13
Xu J, Sun S, Li Q, Yue Y, Li Y, Shao S. A rapid response “turn-on” fluorescent probe for nitroreductase detection and its application in hypoxic tumor cell imaging. Analyst (London), 2015, 140(2): 574–581 https://doi.org/10.1039/C4AN01934B
14
Zhou J, Shi W, Li L H, Gong Q Y, Wu X F, Li X H, Ma H M. A lysosome-targeting fluorescence off-on probe for Imaging of nitroreductase and hypoxia in live cells. Chemistry, an Asian Journal, 2016, 11(19): 2719–2724 https://doi.org/10.1002/asia.201600012
15
Jin C, Zhang Q, Lu W. Selective turn-on near-infrared fluorescence probe for hypoxic tumor cell imaging. RSC Advances, 2017, 7(30): 18217–18223 https://doi.org/10.1039/C7RA01466J
16
Huang B, Chen W, Kuang Y Q, Liu W, Liu X J, Tang L J, Jiang J H. A novel off-on fluorescent probe for sensitive imaging of mitochondria-specific nitroreductase activity in living tumor cells. Organic & Biomolecular Chemistry, 2017, 15(20): 4383–4389 https://doi.org/10.1039/C7OB00781G
17
Zhou Y, Bobba K N, Lv X W, Yang D, Velusamy N, Zhang J F, Bhuniya S. A biotinylated piperazine-rhodol derivative: A ‘turn-on’ probe for nitroreductase triggered hypoxia imaging. Analyst (London), 2017, 142(2): 345–350 https://doi.org/10.1039/C6AN02107G
18
Cui L, Zhong Y, Zhu W, Xu Y, Du Q, Wang X, Qian X, Xiao Y. A new prodrug-derived ratiometric fluorescent probe for hypoxia: High selectivity of nitroreductase and imaging in tumor cell. Organic Letters, 2011, 13(5): 928–931 https://doi.org/10.1021/ol102975t
19
Cai Q, Yu T, Zhu W, Xu Y, Qian X. A turn-on fluorescent probe for tumor hypoxia imaging in living cells. Chemical Communications, 2015, 51(79): 14739–14741 https://doi.org/10.1039/C5CC05518K
20
Chevalier A, Zhang Y, Khdour O M, Kaye J B, Hecht S M. Mitochondrial nitroreductase activity enables selective imaging and therapeutic targeting. Journal of the American Chemical Society, 2016, 138(37): 12009–12012 https://doi.org/10.1021/jacs.6b06229
21
Li Z, He X, Wang Z, Yang R, Shi W, Ma H. In vivo imaging and detection of nitroreductase in zebrafish by a new near-infrared fluorescence off-on probe. Biosensors & Bioelectronics, 2015, 63: 112–116 https://doi.org/10.1016/j.bios.2014.07.024
22
Li Z, Li X, Gao X, Zhang Y, Shi W, Ma H. Nitroreductase detection and hypoxic tumor cell Imaging by a designed sensitive and selective fluorescent probe, 7-[(5-nitrofuran-2-yl)methoxy]-3H-phenoxazin-3-one. Analytical Chemistry, 2013, 85(8): 3926–3932 https://doi.org/10.1021/ac400750r
23
Li Z, Gao X, Shi W, Li X, Ma H. 7-((5-Nitrothiophen-2-yl)methoxy)-3H-phenoxazin-3-one as a spectroscopic off-on probe for highly sensitive and selective detection of nitroreductase. Chemical Communications, 2013, 49(52): 5859–5861 https://doi.org/10.1039/c3cc42610f
24
You X, Li L, Li X, Ma H, Zhang G, Zhang D. A new tetraphenylethylene-derived fluorescent probe for nitroreductase detection and hypoxic-tumor-cell imaging. Chemistry, an Asian Journal, 2016, 11(20): 2918–2923 https://doi.org/10.1002/asia.201600945
25
Ao X, Bright S A, Taylor N C, Elmes R B P. 2-Nitroimidazole based fluorescent probes for nitroreductase; monitoring reductive stress in cellulo. Organic & Biomolecular Chemistry, 2017, 15(29): 6104–6108 https://doi.org/10.1039/C7OB01406F
26
Sedgwick A C, Sun X L, Kim G, Yoon J, Bull S D, James T D. Boronate based fluorescence (ESIPT) probe for peroxynitrite. Chemical Communications, 2016, 52(83): 12350–12352 https://doi.org/10.1039/C6CC06829D
27
Sun X, Xu Q, Kim G, Flower S E, Lowe J P, Yoon J, Fossey J S, Qian X, Bull S D, James T D. A water-soluble boronate-based fluorescent probe for the selective detection of peroxynitrite and imaging in living cells. Chemical Science (Cambridge), 2014, 5(9): 3368–3373 https://doi.org/10.1039/C4SC01417K
28
Gu K Z, Xu Y S, Li H, Guo Z Q, Zhu S J, Zhu S Q, Shi P, James T D, Tian H, Zhu W H. Real-time tracking and in vivo visualization of beta-galactosidase activity in colorectal tumor with a ratiometric near-infrared fluorescent probe. Journal of the American Chemical Society, 2016, 138(16): 5334–5340 https://doi.org/10.1021/jacs.6b01705
29
Li M, Wu X M, Wang Y, Li Y S, Zhu W H, James T D. A near-infrared colorimetric fluorescent chemodosimeter for the detection of glutathione in living cells. Chemical Communications, 2014, 50(14): 1751–1753 https://doi.org/10.1039/c3cc48128j
30
Sedgwick A C, Chapman R S L, Gardiner J E, Peacock L R, Kim G, Yoon J, Bull S D, James T D. A bodipy based hydroxylamine sensor. Chemical Communications, 2017, 53(75): 10441–10443 https://doi.org/10.1039/C7CC05872A
31
Sedgwick A C, Han H, Gardiner J E, Bull S D, He X P, James T D. Long-wavelength fluorescent boronate probes for the detection and intracellular imaging of peroxynitrite. Chemical Communications, 2017, 53(95): 12822–12825 https://doi.org/10.1039/C7CC07845E
32
Matikonda S S, Fairhall J M, Tyndall J D A, Hook S, Gamble A B. Stability, kinetic, and mechanistic investigation of 1,8-self-immolative cinnamyl ether spacers for controlled release of phenols and generation of resonance and inductively stabilized methides. Organic Letters, 2017, 19(3): 528–531 https://doi.org/10.1021/acs.orglett.6b03695
33
Kwon N, Cho M K, Park S J, Kim D, Nam S J, Cui L, Kim H M, Yoon J. An efficient two-photon fluorescent probe for human NAD(P)H: Quinone oxidoreductase (hNQO1) detection and imaging in tumor cells. Chemical Communications, 2017, 53(3): 525–528 https://doi.org/10.1039/C6CC08971B