K. Hałdys, et al.
Bioorganic Chemistry xxx (xxxx) xxxx
found 336.0345. NMR data are in agreement with those reported in the
3
5
+
+
calcd for C
8
ClN
H
9
ClN
3
S (M+H) 214.0206, found 214.0179; m/z calcd
3
5
for C
8
H
8
3
NaS (M+Na) 236.0025, found 236.0028. NMR data
literature [51]
are in agreement with those reported in the literature [40].
4
8
7
.1.1.16. (E)-2-fluorobenzaldehyde thiosemicarbazone (TSC 16). yield
4
9
2
.1.1.10. (E)-3′-chloroacetophenone thiosemicarbazone (TSC 10). yield
1
3%; H NMR (400 MHz, DMSO-d
6
) δ: 7.17–7.23 (m, 2H, ArH),
1
7%; H NMR (600 MHz, DMSO-d
6
) δ: 2.30 (s, 3H, CH ), 7.40–7.45 (m,
3
.37–7.43 (m, 1H, ArH), 8.04 (s, 1H, NH), 8.19 (dt, 1H, J = 1.6 Hz,
H, ArH), 7.89 (dt, 1H, J = 1.8 Hz, J = 7.5 Hz, ArH), 8.08 (t,
J = 7.6 Hz, ArH), 8.24 (s + bs, 2H, CH]N + NH), 11.52 (s, 1H, NH).
J = 1.8 Hz, 1H, ArH), 8.12 (bs, 1H, NH), 8.33 (bs, 1H, NH), 10.26 (s,
+
HRMS (ESI+): m/z calcd for C
8
H
9
3
FN
3
S (M+H) 198.0501, found
1
3
1
H, NH). C NMR (100 MHz, DMSO-d
6
) δ: 14.50, 125.87, 126.67,
+
1
98.0505; m/z calcd for C
H
8 8
FN NaS (M+Na) 220.0321, found
1
29.44, 130.53, 133.94, 140.32, 146.83, 179.54. HRMS (ESI+): m/z
2
20.0322. NMR data are in agreement with those reported in the
3
5
+
calcd for C
9
H
11 ClN
3
S (M+H) 228.0362, found 228.0338; m/z calcd
literature [56,57].
7
9
+
for C
9
H
10 ClN
3
NaS (M+Na) 250.0182, found 250.0196.
4
.1.1.17. (E)-2′-fluoroacetophenone thiosemicarbazone (TSC 17). yield
4
9
7
8
C
C
.1.1.11. (E)-4-chlorobenzaldehyde thiosemicarbazone (TSC 11). yield
1
1
94%; H NMR (400 MHz, DMSO-d
6
) δ: 2.25 (d, JHF = 3.2 Hz, 3H, CH ),
3
0%; H NMR (600 MHz, DMSO-d ) δ: 7.44 (d, 2H, J = 8.7 Hz, ArH),
6
7
2
.13–7.23 (m, 2H, ArH), 7.36–742 (m, 1H, ArH), 7.71–7.76 (dt + bs,
.82 (d, 2H, J = 8.7 Hz, ArH), 8.03 (s, 1H, CH]N), 8.04 (bs, 1H, NH),
H, J = 1.6 Hz, J = 8.0 Hz, ArH + NH), 8.28 (bs, 1H, NH), 10.32 (s,
.20 (bs, 1H, NH), 11.46 (s, 1H, NH). HRMS (ESI+): m/z calcd for
+
+
3
3
5
5
+
1H, NH). HRMS (ESI+): m/z calcd for C
9
H
11FN
3
S (M+H) 212.0658,
8
8
H
9
8
ClN
ClN
3
3
S (M+H) 214.0206, found 214.0206; m/z calcd for
+
found 212.0632; m/z calcd for C
9
H
10FN
3
NaS (M+Na) 234.0477,
H
NaS (M+Na) 236.0025, found 235.9990. NMR data are
found 234.0474.
in agreement with those reported in the literature [27,40,53].
NOTE: product contain ~2% (molar) of the Z isomer. NMR data of
main, E isomer are in agreement with those reported in the literature
[42,44].
4
.1.1.12. (E)-4′-chloroacetophenone thiosemicarbazone (TSC 12). yield
1
9
2%; H NMR (600 MHz, DMSO-d
6
) δ: 2.30 (s, 3H, CH ), 7.43 (d, 2H,
3
J = 8.7 Hz, ArH), 7.98 (d, 2H, J = 8.7 Hz, ArH), 8.01 (bs, 1H, NH), 8.32
(
(
bs, 1H, NH), 10.27 (s, 1H, NH). HRMS (ESI+): m/z calcd for HRMS
4.1.1.18. (E)-3- fluorobenzaldehyde thiosemicarbazone (TSC 18). yield
3
5
+
1
ESI+): m/z calcd for
C
9
H
H
11 ClN
3
S
(M+H) 228.0362, found
+
90%; H NMR (400 MHz, DMSO-d
6
) δ: 7.17 (ddt, 1H, J = 0.8 Hz,
7
9
2
2
28.0389; m/z calcd for C
9
10 ClN
3
NaS (M+Na) 250.0182, found
J = 2.8 Hz, J = 8.8 Hz, ArH), 7.39 (dt, 1H, J = 6.0 Hz, J = 7.6 Hz,
ArH), 7.47 (bd, 1H, J = 7.6 Hz, ArH), 7.80 (ddd, 1H, J = 1.2 Hz,
J = 2.0 Hz, J = 10.4 Hz, ArH), 7.99 (s, 1H, CH]N), 8.12 (bs, 1H,
50.0179. NMR data are in agreement with those reported in the
literature [40].
NH), 8.21, (bs, 1H, NH), 11.47 (s, 1H, NH). HRMS (ESI+): m/z calcd for
+
4
.1.1.13. Mixture
of
(E)-
and
(Z)-4-chlorobenzophenone
C
C
8
8
9
H FN
8
H FN
3
S
3
(M+H)
198.0501, found 198.0509; m/z calcd for
+
1
NaS (M+Na) 220.0321, found 220.0255. NMR data are in
thiosemicarbazone (TSC 13) (in 67:33 M ratio, based on
H
1
NMR). yield 66%; H NMR (400 MHz, DMSO-d
6
) δ: 7.27–7.33 (m,
agreement with those reported in the literature [56].
ArH, E+Z isomers), 7.36 (m [AA‘BB‘ system], m-ArH of 4-ClPh, Z
isomer) and 7.39 (m [AA‘BB‘ system], m-ArH of 4-ClPh, E isomer) [total
integration 4H], 7.55–7.57 (m, 5H, ArH, E+Z isomer), 8.27 (bs, NH, Z
4.1.1.19. (E)-3′-fluoroacetophenone thiosemicarbazone (TSC 19). yield
1
99%; H NMR (400 MHz, DMSO-d ) δ: 2.25 (s, 3H, CH ), 7.16 (ddt, 1H,
6
3
isomer), 8.40 (bs, 2xNH, E isomer) 8.54 (bs, NH, Z isomer), 8.62 (bs,
J = 1.2 Hz, J = 2.8 Hz, J = 8.0 Hz, ArH), 7.37 (dt, 1H, J = 6.0 Hz,
J = 8.0 Hz, ArH), 7.66 (ddd, 1H, J = 1.2 Hz, J = 1.6 Hz, J = 7.6 Hz,
ArH), 7.86 (ddd, 1H, J = 2.4 Hz, J = 1.6 Hz, J = 11.4 Hz, ArH), 8.06
1
NH, E isomer) and 8.83 (bs, NH, Z isomer) [total integration 3H].
H
NMR (600 MHz, DMSO-d ) δ: 7.36–7.39 (m, 2H, ArH, E+Z isomers),
6
7
.42 (d, J = 7.2 Hz, m-ArH of 4-ClPh, Z isomer) and 7.45 (d, J = 8.4 Hz,
(bs, 1H, NH), 8.28 (bs, 1H, NH), 10.22 (s, 1H, NH). HRMS (ESI+): m/z
+
m-ArH of 4-ClPh, E isomer) [total integration 2H], 7.62–7.72 (m, 5H,
ArH, E+Z isomer), 8.32 (bs, NH, Z isomer), 8.44 (bs, NH, E isomer),
calcd for C
9
H11FN
3
S (M+H) 212.0658, found 212.0636; m/z calcd for
+
C H FN NaS (M+Na) 234.0477, found 234.0471. NMR data are in
9
10
3
8
.45 (bs, NH, E isomer) 8.59 (bs, NH, Z isomer), 8.67 (bs, NH, E isomer)
13
agreement with those reported in the literature [42].
and 8.88 (bs, NH, Z isomer) [total integration 3H]. C NMR (100 MHz,
DMSO-d
6
) δ: set of signals of E and Z isomers: 128.11, 128.83, 128.91,
28.94, 129.77, 130.31, 130.46, 130.67, 130.94, 131.13, 131.31,
35.00, 135.14, 135.79, 136.84, 148.35, 148.39, 178.42, 178.76. *
4
6
2
1
.1.1.20. (E)-4-fluorobenzaldehyde thiosemicarbazone (TSC 20). yield
1
1
1
7%; H NMR (400 MHz, DMSO-d
6
) δ: 7.19 (m, 2H, ArH), 7.83 (m,
H, ArH), 7.99 (s + bs, 2H, CH]N + NH), 8.16 (s, 1H, NH), 11.39 (s,
one aromatic CH carbon signal is missing, probably due to coalescence
+
+
H, NH). HRMS (ESI+): m/z calcd for C
8
FN
H
9
FN
3
S (M+H) 198.0501,
3
5
S (M+H)+
and overlaps. HRMS (ESI+): m/z calcd for C14
H
13 ClN
3
found 198.0499; m/z calcd for C
8
H
8
3
NaS (M+Na) 220.0321,
2
90.0519, found 290.0512.
found 220.0323. NMR data are in agreement with those reported in
the literature [53,56]
4
.1.1.14. (E)-3,4-dichlorobenzaldehyde
thiosemicarbazone
(TSC
1
1
4). yield 80%;
H
NMR (600 MHz, DMSO-d
6
) δ: 7.65 (d, 1H,
4
.1.1.21. (E)-4′-fluoroacetophenone thiosemicarbazone (TSC 21). yield
J = 8.4 Hz, ArH), 7.72 (dd, 1H, J = 1.2 Hz, J = 8.4 Hz, ArH), 8.00 (s,
1
9
0%; H NMR (600 MHz, DMSO-d
6
) δ: 2.30 (s, 3H, CH ), 7.20 (bt, 2H,
3
1
H, CH]N), 8.25 (s, 2H, ArH + NH), 8.29 (bs, 1H, NH). 11.57 (s, 1H,
3
5
+
+
J = 8.1 Hz, ArH), 7.98 (bs, 1H, NH), 8.00 (d, 2H, J = 8.7 Hz, ArH), 8.28
NH). HRMS (ESI+): m/z calcd for C
8
H
8
Cl
2
N
3
S (M+H) 247.9816,
3
5
(bs, 1H, NH), 10.22 (s, 1H, NH). HRMS (ESI+): m/z calcd for HRMS
found 247.9817; m/z calcd for C
8
H
7
Cl
2
N NaS (M+Na) 269.9635,
3
+
(
ESI+): m/z calcd for C
9
H
11FN
3
S (M+H) 212.0658, found 212.0641;
found 269.9625. NMR data are in agreement with those reported in the
literature [43,55].
+
m/z calcd for C
9
H
10FN
3
NaS (M+Na) 234.0477, found 234.0484.
NMR data are in agreement with those reported in the literature [28].
4
.1.1.15. (E)-10-Chloro-9-anthraldehyde
thiosemicarbazone
(TSC
1
1
5). yield 77%; H NMR (600 MHz, DMSO-d
6
) δ: 7.73 (m, 2H, ArH),
4.2. Enzyme preparation
7
.78 (m, 3H, NH + ArH), 8.35 (bs, 1H, NH), 8.57 (d, 2H, J = 9.0 Hz,
ArH), 8.52 (d, 2H, J = 9.0 Hz, ArH), 9.26 (s, 1H, CH]N), 11.72 (s, 1H,
The enzyme was isolated from Agaricus bisporus purchased from a
local supplier in Wroclaw. The full description of enzyme preparation is
included in electronic Supplementary Materials.
3
5
+
+
NH). HRMS (ESI+): m/z calcd for C16
H
13 ClN
3
S (M+H) 314.0519,
3
5
found 314.0560; m/z calcd for C16
H
12 ClN
3
NaS (M+Na) 336.0338,
9