A simple umbelliferone based fluorescent probe for the detection of nitroreductase

Abstract

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.

Acknowledgements

We would like to thank the EPSRC and the University of Bath for funding. TDJ wishes to thank the Royal Society for a Wolfson Research Merit Award. ACS thanks the EPSRC for his studentship. RBPE acknowledges support funding from Maynooth University. NMR characterisation facilities were provided through the Chemical Characterisation and Analysis Facility (CCAF) at the University of Bath (www.bath.ac.uk/ccaf). All data supporting this study are provided as supplementary information accompanying this paper.

Electronic Supplementary Material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s11705-017-1697-0 and is accessible for authorized users.

References

1 Brown J M, Wilson W R. Exploiting tumour hypoxia in cancer treatment. Nature Reviews. Cancer, 2004, 4(6): 437–447

2 Wilson W R, Hay M P. Targeting hypoxia in cancer therapy. Nature Reviews. Cancer, 2011, 11(6): 393–410

3 Denny W A. Prodrug strategies in cancer therapy. European Journal of Medicinal Chemistry, 2001, 36(7-8): 577–595

4 Elmes R B P. Bioreductive fluorescent imaging agents: Applications to tumour hypoxia. Chemical Communications, 2016, 52(58): 8935–8956

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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