Arch. Pharm. Chem. Life Sci. 2016, 349, 1–9
CrAsH-EDT Photoaffinity Probes
ARC HH PHARM
2
Archiv der Pharmazie
was purified by flash column chromatography (silica gel,
J ¼ 8.0, 1.4 Hz, 1H), 7.96 (s, 1H), 7.79 (d, J ¼ 2.7 Hz, 1H), 7.22 (d,
J ¼ 8.0 Hz, 1H), 7.07 (dd, J ¼ 9.2, 2.7 Hz, 1H), 6.80 (d, J ¼ 9.2 Hz,
1H), 6.51 (d, J ¼ 8.8 Hz, 2H), 6.44 (d, J ¼ 8.8 Hz, 2H), 6.34 (t,
J ¼ 5.7 Hz, 1H), 3.65–3.36(m, 10H), 3.24(dd, J ¼ 12.2, 6.9 Hz, 2H),
toluene/EtOAc, 4:1) to furnish 0.04 g (86%) of 6 as yellow
1
solid: H NMR (400 MHz, CDCl
3
) d 9.98 (s, 2H), 8.81 (d,
J ¼ 0.8 Hz, 1H), 8.44 (dd, J ¼ 8.1, 1.5 Hz, 1H), 7.40 (d, J ¼ 8.1 Hz,
1
3
1
H), 6.63 (d, J ¼ 8.8 Hz, 2H), 6.57 (d, J ¼ 8.8 Hz, 2H), 3.84–3.38
3
1.78–0.94 (m, 8H); C NMR (100 MHz, CDCl ) d 168.3, 165.5,
(
m, 8H), 2.97 (s, 4H); HRMS (ESI) calcd. for C29
22
H O
9
S
4
As
2
162.9, 154.6, 152.3, 143.1, 136.7, 134.7, 131.4, 130.4, 128.1,
127.5, 127.0, 124.5, 122.6, 115.8, 115.3, 114.8, 112.4, 109.6, 82.7,
þ
(
MþH ) 805.8609, found 805.8616.
4
3.3, 43.0, 40.0, 29.5, 29.3, 28.6, 26.5, 26.4; HRMS (ESI) calcd.
þ
4
-Azido-1-fluoro-2-nitrobenzene (8) [30]
for C37
H N O S As
35 6 8 4 2
(MþH ) 968.9831, found 968.9822.
4
-Fluoro-3-nitroaniline (7) (2.00 g, 12.80 mmol) was dissolved
in a mixture of concentrated hydrochloric acid (12 mL) and
water (2 mL) by heating (50°C). The resulting brown mixture
tert-Butyl (6-(3-(3-methyl-3H-diazirin-3-yl)propanamido)-
hexyl)carbamate (12)
was cooled to ꢁ70°C and then NaNO
2
(0.96 g, 13.84 mmol)
dissolved in water (2 mL) was slowly added and then stirred at
the same temperature for 10 min. The reaction mixture was
To a stirred solution of commercially available 11 (0.02 g,
0.10 mmol) in THF was added commercially available amine 10
(0.03 g, 0.15 mmol) at ambient temperature and the reaction
mixture was stirred for 3 h. After completion of the reaction
(monitored by TLC), THF was removed in vacuo and the
residue was purified by column chromatography (silica gel,
rapidly filtered at ꢁ70°C. At this temperature NaN
3
(0.88 g,
1
3.58 mmol), dissolved in water (3.2 mL) was added. An
orange to tan-colored precipitate was observed. The reaction
was stirred for a few minutes and then ice-cold water was
added to the reaction mixture. The solid was filtered and
hexanes/EtOAc, 4:1) to provide (0.03 g) 97% of 12 as a white
1
solid: mp 97.4–100.6°C; H NMR (400 MHz, CDCl
3
) d 5.64 (bs,
washed with water and dried under vacuum to provide 2.29 g
1H), 4.53 (bs, 1H), 3.23 (dd, J ¼ 13.0, 6.7 Hz, 2H), 3.11 (d,
1
(
98%) of 8 as a tan-colored solid: mp 53.7–54.8°C; H NMR
J ¼ 5.9 Hz, 2H), 1.98 (t, J ¼ 7.7 Hz, 2H), 1.76 (t, J ¼ 7.7 Hz, 2H),
1
3
(
2
400 MHz, CDCl
3
) d 7.73 (dd, J ¼ 6.1, 2.6 Hz, 1H), 7.36–7.25 (m,
3
1.58–1.24 (m, 17H), 1.03 (s, 3H); C NMR (100 MHz, CDCl )
13
H); C NMR (100 MHz, CDCl ) d 153.7, 151.1, 137.1 (d,
3
d 171.22, 156.11, 79.13, 40.09, 39.17, 30.69, 30.06, 29.99,
J ¼ 3.6 Hz), 125.7 (d, J ¼ 8.3 Hz), 119.9 (d, J ¼ 22.7 Hz), 116.1 (d,
29.34, 28.41, 26.06, 25.91, 25.46, 19.94.
þ
J ¼ 2.4 Hz); HRMS (ESI) calcd. for C12
H
N
4 4
O
2
(MþH ) 183.0318,
found 183.0307.
N-(6-Aminohexyl)-3-(3-methyl-3H-diazirin-3-yl)-
propanamide (13)
1
N -(4-Azido-2-nitrophenyl)hexane-1,6-diamine (9)
To a solution of 12 (0.03 g, 0.10 mmol) in dichloromethane
(2 mL) was added trifluoroacetic acid (1 mL) at 0°C. After
stirring for 1 h at 0°C, the reaction mixture was concentrated
in vacuo and the residue was dissolved in EtOAc and washed
with saturated aqueous sodium bicarbonate solution and
extracted with EtOAc. The combined organic layers were
dried over Na SO and dried to furnish 13 (0.02 g) in 87%
2 4
yield, which was used for the preparation of probe 2 without
further purification.
To a stirred solution of 1-fluoro-2-nitro-4-azidobenzene (8)
(
0.18 g, 0.99 mmol) in MeOH (2 mL) was added hexamethyl-
enediamine (0.58 g, 5.00 mmol) in MeOH (20 mL) at room
temperature and the reaction mixture was stirred for 12 h.
The solvent was removed in vacuo and EtOAc was added
to the residue, unreacted diamine precipitated, and was
removed by filtration and the filtrate was extracted with
EtOAc and washed with H
and concentrated in vacuo to furnish 0.22 g (80%) of 9 as
brown-colored solid: mp 59.1–60.8°C; H NMR (400 MHz,
2
O (4 ꢀ 25 mL), dried over Na
2 4
SO
1
0
0
0
0
4 ,5 -Di(1,3,2-dithiarsolan-2-yl)-3 ,6 -dihydroxy-N-(6-(3-(3-
methyl-3H-diazirin-3-yl)propanamido)hexyl)-3-oxo-3H-
CDCl
3
) d 8.03 (s, 1H), 7.86 (d, J ¼ 2.7 Hz, 1H), 7.13 (dd, J ¼ 9.2,
0
2
.7 Hz, 1H), 6.87 (d, J ¼ 9.2 Hz, 1H), 3.36–3.22 (m, 2H), 2.75 (t,
spiro[isobenzofuran-1,9 -xanthene]-5-carboxamide
1
3
J ¼ 7.0 Hz, 2H), 1.83–1.64 (m, 2H), 1.62–1.31 (m, 6H); C NMR
100 MHz, CDCl ) d 143.3, 131.4, 128.2, 127.7, 115.9, 115.5,
3.1, 40.5, 30.4, 28.8, 26.6, 26.3; HRMS (ESI) calcd. for
(diazirine-TRAP) (2)
(
4
C
3
To a stirred solution of NHS ester 6 (0.05 g, 0.06 mmol) in THF
(5 mL) was added 13 (0.01 g, 0.07 mmol) at room temperature.
After being stirred for 3 h, THF was removed in vacuo and the
residue was purified by column chromatography (silica gel,
þ
H
12 19
N
6
O
2
(MþH ) 279.1569, found 279.1566.
0
0
N-(6-(5-Azido-2-nitrophenylamino)hexyl)-4 ,5 -di(1,3,2-
EtOAc/toluene 5:1) to provide probe 2 (0.03 g) as an orange
0
0
1
dithiarsolan-2-yl)-3 ,6 -dihydroxy-3-oxo-3H-spiro-
solid in 53% yield: H NMR (400 MHz, CDCl
3
) d 9.90 (s, 2H),
0
[
isobenzofuran-1,9 -xanthene]-5-carboxamide (azide-
8.37 (d, J ¼ 1.6 Hz, 1H), 8.22 (dd, J ¼ 8.0, 1.6 Hz, 1H), 7.26
(d, J ¼ 8.1 Hz, 1H), 6.83 (s, 1H), 6.64–6.39 (m, 4H), 5.63
(t, J ¼ 6.0 Hz, 1H), 3.74–3.37 (m, 10H), 3.24 (q, J ¼ 6.6 Hz,
2H), 1.97 (dd, J ¼ 8.5, 6.6 Hz, 2H), 1.75 (dd, J ¼ 8.6, 6.7 Hz, 2H),
TRAP) (1)
To a stirred solution of NHS ester 6 (0.02 g, 0.03 mmol) in THF
(
5 mL) was added 9 (0.01 g, 0.05mmol) at room temperature.
1
3
After stirring for 3 h, THF was removed in vacuo and the residue
was purified by column chromatography (silica gel, EtOAc/
3
1.63 (m, 8H), 0.99 (s, 3H); C NMR (100 MHz, CDCl ) d 170.56,
167.41, 164.69, 162.01, 153.61, 151.46, 135.98, 133.83, 129.60,
126.16, 123.56, 122.07, 114.00, 111.60, 108.94, 82.23, 38.60,
37.86, 30.91, 29.71, 28.98, 28.69, 28.46, 28.19, 26.29, 21.68;
toluene 5:1) to yield 0.02 g (77%) of probe 1 as an orange solid:
1
3
H NMR (400 MHz, CDCl ) d 9.86 (s, 2H), 8.23 (s, 1H), 8.14 (dd,
ß 2016 The Authors. Archiv der Pharmazie Published by Wiley-VCH Verlag GmbH & Co. KGaA.
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