Hybrids of a 9-anthracenyl moiety and fluorescein as … · 2019-11-01 · treated with InPpa (0.1...
19
S1 Supporting Information Hybrids of a 9-anthracenyl moiety and fluorescein as chemodosimeters for detection of singlet oxygen in live cells Serghei Chercheja, Steffen Daum, Honggui Xu, Frank Beierlein, and Andriy Mokhir* Content Outline of synthesis of probes 8d and 8e p. S2 NMR spectra of intermediates and new singlet oxygen probes p. S3-S14 Calculation of energies of highest occupied orbitals (HOMOs) of simple models mimicking probes 4, 8a-e p. S15-S16 1 H NMR spectroscopic monitoring of the reaction of probe 4 with 1 O2. p. S17 Detection of 1 O2 in prostate cancer cells DU-145 monitored by using fluorescence microscopy in combination with probe 4. p. S18 References p. S19 Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry. This journal is © The Royal Society of Chemistry 2019
Transcript of Hybrids of a 9-anthracenyl moiety and fluorescein as … · 2019-11-01 · treated with InPpa (0.1...
S1
Supporting Information
Hybrids of a 9-anthracenyl moiety and fluorescein as chemodosimeters for detection of singlet oxygen in live cells
Serghei Chercheja, Steffen Daum, Honggui Xu, Frank Beierlein, and Andriy Mokhir*
Content
Outline of synthesis of probes 8d and 8e p. S2
NMR spectra of intermediates and new singlet oxygen probes p. S3-S14
Calculation of energies of highest occupied orbitals (HOMOs) of
simple models mimicking probes 4, 8a-e
p. S15-S16
1H NMR spectroscopic monitoring of the reaction of probe 4 with
1O2.
p. S17
Detection of 1O2 in prostate cancer cells DU-145 monitored by
using fluorescence microscopy in combination with probe 4.
p. S18
References p. S19
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2019
S2
Outline of synthesis of probes 8d and 8e
Scheme S1.Synthesis of probe 8d: a) chloromethyl methyl sulfide, NaH, DMSO, rt, 24
h, 7%; (b) SO2Cl2, CH2Cl2, 22 °C, 1 h; (c) compound 6, t-BuOK, DMF, rt, 24 h; d) LiOH,
H2O/THF, 22 °C, 24 h, (2% over 2 steps).
Scheme S2. Synthesis of probe 8e: (a) according to (1). Whitton, A. J.; Kumberger,
O.; Müller, G.; Schmidbaur, H. Chem. Ber. 1990, 123, 1931-1939 and (2) Whitton, A.
J.; Kumberger, O.; Müller, G.; Schmidbaur, H. Chem. Ber. 1990, 123, 1931-1939: t-
BuCl, AlCl3, CHCl3, reflux, 24 h, 34%; (b) according to U. Müller, V. Enkelmann, M.
Adam and K. Müllen, Chemische Berichte 1993, 126, 1217-1225: (NH4)2Ce(NO3)6,
THF/H2O, rt, 2 h, 64%; (c) according to Müller, U.; Enkelmann, V. ; Adam, M.; Müllen,
K. Chem. Ber. 1993, 126, 1217-1225 and Müller, U.; Enkelmann, V. ; Adam, M.;
Müllen, K. Chem. Ber. 1993, 126, 1217-1225: Sn, AcOH, HCl, 81%; (d) Chloromethyl
methyl sulfide, NaH, DMSO, rt, 18 h, 5%; (e) SO2Cl2, CH2Cl2, rt, 1 h; (f) compound 6,
t-BuOK, DMF, rt, 24 h, 11%; (g) LiOH, H2O/THF, rt, 24 h, 15%.
S3
NMR spectra of intermediates and new singlet oxygen probes 4, 8b-8e
9 8 7 6 5 4 3 2 1 0
6.0
0
2.9
7
1.0
0
1.8
8
1.9
8
1.0
1
1.0
2
2.0
5
1.0
3
8.0
38.0
07.6
87.6
7
7.6
57.6
17.5
97.2
77.1
7 6.7
6 6.6
76.6
66.6
66.5
76.5
36.5
36.5
06.5
0
3.7
9
1.6
4
Figure S1. 1H NMR spectrum of compound 6.
200 180 160 140 120 100 80 60 40 20 0
174.6
170.1
158.7 1
57.3
152.6
152.2
135.1
129.8
129.2
128.9
126.7
125.1
124.2
115.0
112.7
112.4
110.6 105.7
103.1
79.5
77.4 77.0
76.6
52.8
25.4
25.2
Figure S2. 13C NMR spectrum of compound 6.
S4
9 8 7 6 5 4 3 2 1 0
6.0
9
2.9
4
2.9
8
1.0
6
2.0
4
1.0
3
4.0
3
2.0
6
4.9
8
1.0
08.4
3
8.0
98.0
78.0
48.0
37.7
17.7
1 7.6
67.5
17.5
17.4
87.4
87.4
57.4
37.2
77.2
1 6.7
3 6.7
26.7
16.6
16.5
16.5
06.4
96.4
86.4
7
3.7
3
1.5
9
Figure S3. 1H NMR spectrum of compound 7a.
200 180 160 140 120 100 80 60 40 20 0
174.2
169.2
161.6
157.2
152.7
152.6
151.9
144.7
135.0 132.3
128.8
128.4
126.2
125.8
124.3
122.0
115.0
112.5
112.2
105.5
103.0
82.9 7
9.4
77.4
77.0
76.6
52.7
25.4
25.1
Figure S4. 13C NMR spectrum of compound 7a.
S5
9 8 7 6 5 4 3 2 1 0
6.2
0
0.8
7
5.0
5
1.0
9
4.0
1
2.1
3
0.9
8
1.8
9
1.9
1
0.9
18.6
18.2
18.2
08.1
88.0
87.7
47.5
87.5
77.5
6 7.5
57.5
47.5
37.5
27.3
4 6.8
2 6.6
76.6
66.6
46.5
56.5
5
1.5
9
Figure S5. 1H NMR spectrum of probe 4.
220 200 180 160 140 120 100 80 60 40 20 0
206.7
206.5
206.3
175.1
169.5
162.7
158.8
153.7
153.5
152.8
145.6
136.5 133.5
129.7
127.7
127.0
125.7
122.7
116.3
113.3
106.3 103.7
82.9
80.2
30.5
30.3
30.1
29.9
29.7
29.5
29.3
25.6
25.6
Figure S6. 13C NMR spectrum of probe 4.
S6
9 8 7 6 5 4 3 2 1 0
4.0
6
2.0
4
2.0
08.6
4
8.6
38.6
3 8.5
98.5
8
7.9
27.9
2
7.9
17.9
1
7.8
77.8
67.7
17.7
07.6
67.6
5
7.2
7
Figure S7. 1H NMR spectrum of 9-bromo-10-nitroanthracene.
200 180 160 140 120 100 80 60 40 20
129.9
129.0
128.2
128.0
126.4
122.7
121.7
77.4 7
7.0
76.6
Figure S8. 13C NMR spectrum of 9-bromo-10-nitroanthracene.
S7
9 8 7 6 5 4 3 2 1 0
6.3
7
2.9
6
2.9
3
1.0
0
1.9
9
0.9
6
1.9
74
.04
2.9
1
2.0
08.1
38.1
0 8.0
27.9
97.7
27.7
2
7.6
87.6
8 7.6
7 7.6
57.6
5
7.5
57.5
37.5
37.2
16.7
76.7
46.6
36.5
36.4
96.4
9
3.7
4
1.5
9
Figure S9. 1H NMR spectrum of compound 7b.
200 180 160 140 120 100 80 60 40 20 0
174.1
169.1
160.9
157.2
152.6
152.5
151.8
147.3
142.0
135.0
129.8
129.2
127.0
123.9
122.4
121.7
115.1
113.3
112.1
112.0
105.4 103.1
82.6
79.4
77.4
77.0
76.6
52.6
25.3
25.1
Figure S10. 13C NMR spectrum of compound 7b.
S8
9 8 7 6 5 4 3 2 1 0
5.9
8
1.0
0
1.0
4
2.1
6
1.7
6
1.0
0
2.0
6
4.0
6
3.0
4
2.0
38.1
38.1
08.0
0 7.9
77.6
77.6
77.6
67.5
4 7.2
77.2
07.1
8
6.7
66.7
3
6.6
5 6.6
46.6
06.5
46.5
3
1.6
0
Figure S11. 1H NMR spectrum of compound 8b.
200 180 160 140 120 100 80 60 40 20 0
178.1
169.2 161.0
156.7
152.6
151.8
147.3
142.0 135.1
129.9
129.3
127.0
123.9
122.4
121.8
116.0
113.3
112.8
112.0
106.7
103.1
82.6
79.4
77.4 77.0
76.6
25.3 25.0
Figure S12. 13C NMR spectrum of compound 8b.
S9
9 8 7 6 5 4 3 2 1 0
5.9
7
2.7
2
0.8
7
3.0
1
1.9
5
1.0
0
3.7
2
2.2
1
1.0
0
1.9
1
1.9
7
0.8
69.5
6 8.2
78.2
48.0
78.0
48.0
4
7.9
77.7
97.6
97.5
57.5
27.5
17.5
17.4
87.4
87.2
76.8
36.8
3 6.6
86.6
56.6
36.4
86.4
7
2.4
2
2.0
62.0
52.0
4
1.5
5
Figure S13. 1H NMR spectrum of compound 8c.
200 180 160 140 120 100 80 60 40 20 0
206.3
175.1
170.8
170.7
169.3
162.5
158.7
153.5
153.3
152.6
144.8
136.3
131.0
130.9 130.3
127.7
127.4
127.2
125.5
125.3
122.8
114.3 1
13.2
106.1
103.6
82.9
80.2
30.7
30.4
30.2
29.9
29.7
29.4
29.1
25.7
25.6
23.3
Figure S14. 13C NMR spectrum of compound 8c.
S10
Figure S15. 1H NMR spectrum of 9-(methylthiomethoxy)anthracene.
Figure S16. 13C NMR spectrum of 9-(methylthiomethoxy)anthracene.
S11
Figure S17. 1H NMR spectrum of compound 8d.
Figure S18. 13C NMR spectrum of compound 8d.
S12
Figure S19. 1H NMR spectrum of 2,6-di-tert-butyl-9-(methylthiomethoxy)anthracene.
Figure S20. 13C NMR spectrum of 2,6-di-tert-butyl-9-(methylthiomethoxy)anthracene.
S13
9 8 7 6 5 4 3 2 1 0
9.0
7
8.9
2
5.9
4
3.1
4
1.8
8
1.1
3
2.0
7
1.0
5
1.0
3
1.0
9
0.9
7
4.1
1
2.0
4
1.0
0
1.8
48.4
38.4
08.1
2 8.0
78.0
77.6
8 7.6
57.6
4
7.3
07.2
76.8
96.8
9 6.8
2
6.7
36.7
26.7
06.5
86.5
7
5.9
5
3.8
1
1.6
6
1.4
9
1.2
4
Figure S21. 1H NMR spectrum of compound 7e.
180 160 140 120 100 80 60 40 20 0
174.3
169.3
159.1
157.3
152.9
152.6
152.1
149.2
148.6
135.0
129.8
129.5
128.9
126.8
126.5
125.6
125.1
124.5
124.0
122.1
119.4
117.0
115.0
112.8
105.8
103.5
96.4
82.8 79.5
77.3
77.0
76.7
52.7
35.3
35.0
30.9
30.6
25.4
25.3
Figure S22. 13C NMR spectrum of compound 7e.
S14
9 8 7 6 5 4 3 2 1 0
9.3
9
8.9
7
6.4
4
1.9
3
1.1
3
2.1
2
1.0
8
1.0
8
2.1
0
3.0
8
1.1
8
1.1
0
1.0
0
1.0
4
1.9
08.4
18.4
08.3
88.2
88.1
58.1
57.8
17.7
97.7
87.7
87.7
67.3
37.3
26.9
16.8
86.8
46.8
36.7
86.7
3
6.1
66.1
56.1
3
2.0
62.0
62.0
52.0
42.0
4
1.6
5
1.4
8
1.2
2
Figure S23. 1H NMR spectrum of probe 8e.
200 180 160 140 120 100 80 60 40 20 0
206.2
169.5 160.1
158.8
153.9
153.6
150.8
150.0
149.3
136.3
131.1
130.7
129.9
128.0
126.7
125.7
125.1
125.1
123.6
119.8
118.4
114.6
113.9
104.3
97.4 83.0
80.4
36.1
35.7
31.2
31.1
30.7
30.4
30.2
29.9
29.7
29.4
29.1
25.8
25.7
Figure S24. 13C NMR spectrum of probe 8e.
S15
Calculation of energies of highest occupied orbitals (HOMOs) of simple models
mimicking probes 4, 8a-e
Geometry optimizations were performed using the B3LYP1 hybrid density functional
and the 6-31G**2-11 basis set with Gaussian16.12 Frequency calculations of the
optimized structures were used to confirm stationary points (minima). Checkpoint files
were converted to formatted checkpoint files using the formchk command and cube
files containing MO data were obtained with the cubegen command from the Gaussian
suite. Visualization of the HOMO isosurfaces was performed with vmd 1.9.3,13, 14 using
orbital isovalues of -0.02 (yellow) and 0.02 (blue).
Table S1.
Gaussian 16 (B3LYP/6-31G**)
Probe Model structure HOMO energy
[eV] HOMO
Relative HOMO-
energies [eV]
Reference
-4.77
reference, E=0
8e
-5.20
-0.43
8d
-5.20
-0.43
S16
4
-5.27
-0.50
8c
-5.28
-0.51
8b
-5.78
-1.01
Intensity of green color in the last column of this table indicates the reactivity of the
corresponding anthracene derivative with 1O2 as predicted based on energies of the
HOMOs.
S17
Figure S25. 1H NMR spectroscopic monitoring conversion of probe 4 (5 mM) to
anthraquinone AQ in the presence of photochemically generated 1O2. 1O2 was
generated upon irradiation of photosensitizer pyropheophorbide a (Ppa, 0.1 eq, 0.5
mM) with red light (650 nm) for 1 h and 14 h. Spectrum of reference compound AQ is
shown as a red trace. Peaks of AQ appearing upon the transformation of probe 4 are
indicated with dotted, black lines.
2
1H-NMR monitoring
Figure XX. 1H NMR spectroscopic monitoring conversion of probe 4 (5 mM) to anthraquinone AQ in the
presence of photochemically generated 1O2.1O2 was generated upon irradiation of photosensitizer Ppa (0.1
eq, 0.5 mM) with red light (650 nm) for 1 h and 14 h. Spectrum of reference compound AQ is shown as a
red trace. Peaks of AQ appearing upon the transformation of probe 4 are indicated with dotted, black lines.
AQ: reference
Probe 4 + Ppa/irradiated (1 h)
Probe 4 + Ppa/irradiated (14 h)
Probe 4 + Ppa/in dark (1 h)
Probe 4 + Ppa/in dark (14 h)
Chemical Shift
S18
Figure S26. Fluorescence microscopy images combined with optical image of DU-
145 cells before (left column) and after (right column) irradiation under red channel.
A: Cells treated with a carrier DMF; B: Cells treated with probe 4 (1 µM); C: Cells
treated with InPpa (0.1 µM); C: Cells treated with probe 4 (1 µM ) and InPpa (0.1
µM).
3
Fluorescence microscopy
Figure XX. Fluorescence microscopy images combined with optical image of DU-145 cells before (left
column) and after (right column) irradiation under red channel. A: Cells treated with a carrier DMF; B: Cells
treated with probe 4 (1 µM); C: Cells treated with InPpa (0.1 µM); C: Cells treated with probe 4 (1 µM ) and
InPpa (0.1 µM).
S19
References
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