M. Wright, A. D. Miller / Bioorg. Med. Chem. Lett. 14 (2004) 2813–2816
Table 1. Characterisation details of fluorescent Ap4A analogues
2815
Derivative
(ES) [M ꢁ H]ꢁ (m=z) Overall yield (%) Amax (nm)
Imax (nm) 1H and 31P (d ppm)
mant–Ap4A
968.1
85
357
446
1H: 8.5 (1H, s, 8-H-Ad), 8.4 (1H, s, 8-H-Ad [mant]),
8.1 (1H, s, 2-H-Ad), 8.0 (1H, s, 2-H-Ad [mant]), 7.9–
7.8 (1H, d m, 1-H-mant), 7.4–7.3 (1H, m, 3-H-mant),
6.7–6.5 (1H, m, 2-H-mant), 6.2–6.1 (1H, d m, 4-H-
mant)a, 5.9 (2H, m, 10-H-rib)
31P: ꢁ11.2 (2P, m, a-P), ꢁ23.0 (2P, m, b-P)
1H: 8.6 (1H, s, 8-H-Ad), 8.5 (1H, s, 8-H-Ad[mant]),
8.2 (1H, s, 2-H-Ad), 8.1 (1H, s, 2-H-Ad[mant]), 8.0–
7.8 (1H, d m, 1-H-mant), 7.5–7.4 (1H, m, 3-H-mant),
6.8–6.6 (1H, m, 2-H-mant), 6.3–6.1 (1H, d m, 4-H-
mant), 5.9 (2H, m, 10-H-rib), 2.6–2.4 (2H, t, O-CH2-
O, JH–P 20.8 Hz)
mant–AppCH2ppA
966.1
85
357
446
31P: ꢁ10.6 (2P, m, a-P), 8.0 (2P, m, b-P)
1H: 9.1 (ꢂ1H, s, H-TNP), 8.4 (1H, s, 8-H-Ad), 8.35
(1H, s, 8-H-Ad[TNP]), 8.1 (1H, s, 2-H-Ad), 8.05 (1H,
s, 2-H-Ad[TNP]), 6.2 (1H, m, 10-H-rib[TNP]), 5.85
(1H, m, 10-H-rib)
31P: ꢁ11.2 (2P, m, a-P), ꢁ22.7 (2P, m, b-P)
1H: 8.6 (ꢂ1H, s, H-TNP), 8.4 (1H, s, 8-H-Ad), 8.35
(1H, s, 8-H-Ad[TNP]), 8.1 (1H, s, 2-H-Ad), 8.05 (1H,
s, 2-H-Ad[TNP]), 6.4 (1H, m, 10-H-rib[TNP]), 5.95
(1H, m, 10-H-rib), 2.6–2.4 (2H, t, O–CH2–O, JHꢁP
21 Hz)
TNP–Ap4A
1046.0
1043.8
80
80
412
412
546b
546
TNP–AppCH2ppA
31P: ꢁ10.4 (2P, m, a-P), 9.3 (2P, m, b-P)
Mass spectrometry was carried out using an electrospray Bruker Esquire 3000 machine set to 100% fragmentation strength. Samples were applied in
1:1 acetonitrile/water containing 0.1% acetic acid. UV–vis and fluorescence spectra were obtained on an Ultraspec 4000 UV–vis spectrometer and a
Shimadzu RF-5301PC fluorescence spectrophotometer, respectively. Proton and phosphorous NMR spectra were recorded on a 400 MHz Bruker
Ultrashield with samples in D2O.
a Mant protons numbered anticlockwise with respect to Scheme 1.
b Emission maxima are for aqueous solution, TNP derivatives show a shift to 535 nm in 50% ethanol/water.
to mention the identification of polyphosphate-selective
receptors/binding proteins located inside cells.
authors thank Jim Pullen for assistance with the 31P
NMR spectroscopy.
NH2
O
N
O
O
O
O
O
References and notes
P
P
P
N
N
O
O
X
RO
O
N
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NO2
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2
6
7
8
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X = CH2; R = Adenosine 5'-monophosphoryl
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Acknowledgements
M.W. would like to acknowledge IC-Vec for personal
support and we would like to thank IC-Vec and the
Mitsubishi Chemical Corporation for their support of
the Imperial College Genetic Therapies Centre. The
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