Thomas Efferth. 1. *. 1 Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Mainz, Germany.
Supporting Information
Treatment of Multidrug-Resistant Leukemia Cells by Novel Artemisinin-, Egonol-, and Thymoquinone-Derived Hybrid Compounds
Lisa Gruber1, Sara Abdelfatah1, Tony Fröhlich2, Christoph Reiter2, Volker Klein2, Svetlana B. Tsogoeva2, Thomas Efferth1* 1
2
Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Mainz, Germany Department of Chemistry and Pharmacy, Organic Chemistry Chair I and Interdisciplinary Center for Molecular Materials (ICMM), University of ErlangenNuremberg, Erlangen, Germany
S1
Figure S1: Chemical structures of new artemisinin-based hybrids, dimers and trimers studied in this work.
S2
Purity Data TF 19: MS (ESI): m/z = 1064 [M+Na]+. HRMS (ESI): m/z calcd. for [C56H80NaO18]+: 1063.5237, found: 1063.5244. Anal. calcd. for C56H80O18: C, 64.60; H, 7.74; Found: C, 64.58; H, 7.84. TF 26: MS (ESI): m/z = 948 [M+Na]+. HRMS (ESI): m/z calcd. for [C52H76NaO14]+: 947.5127, found: 947.5151. Anal. calcd. for C52H76O14: C, 67.51; H, 8.28; Found: C, 67.34; H, 8.34. REI 213: MS (ESI): m/z = 657 [M+Na]+. HRMS (ESI): m/z calcd. for [C36H42NaO10]+ 657.2670, found 657.2682. Anal. calcd. for C36H42O10: C, 68.12; H, 6.67; Found: C, 68.20; H, 7.02. REI 220+226: MS (MALDI-TOF, without matrix): m/z = 485 [M]+. HRMS (ESI): m/z calcd. for [C29H28NO6]+ 486.1911, found 486.1897. Anal. calcd. for C29H27NO6: C, 71.74; H, 5.61; N, 2.88; Found: C, 71.39; H, 5.57; N, 2.81. REI 234: MS (MALDI-TOF, without matrix): m/z = 592 [M]+. HRMS (ESI): m/z calcd. for [C34H40NaO9]+ 615.2565, found 615.2588. Anal. calcd. for C34H40O9: C, 68.90; H, 6.80; Found: C, 68.99; H, 6.97. REI 259: MS (ESI): m/z = 705 [M+Na]+, 1387 [2M+Na]+. HRMS (ESI): m/z calcd. for [C38H42FeNaO8]+ 705.2122, found 705.2128. Anal. calcd. for C38H42FeO8: C, 66.86; H, 6.20; Found: C, 66.86; H, 6.07; N, 0.08. HH 3: MS (ESI): m/z = 434 [M]. Anal. calcd. for: [C24H26FeO4]: C, 66.37; H, 6.03. Found: C, 66.70; H, 6.23. VK 3: MS (ESI): m/z = 809 [M+Na]+. HRMS (ESI): m/z calcd. for [C43H54FeO10Na]+: 809.29536, found: 809.29594. Anal. calcd. for C43H54FeO10: C, 65.65; H, 6.92, Found: C, 65.62.17; H, 7.18.
S3
2.0
6.13 6.81 1.31
14.25 12.83 1.64 10.04 2.53 7.40
3.0 9.75
4.0 2.06 3.93
5.0 0.95
4.97
6.0 1.99
2.00
7.0 1.0
0.94 0.92 0.83 0.80
1.05
1.16
1.53 1.39 1.32 1.25 1.24 1.23 1.22
1.79 1.59
1.91
2.04 1.96
2.27
2.60
2.65
2.75
3.01 2.99 2.97 2.94 2.92 2.90 2.87
4.03
4.13
5.39
5.74
5.77
1
H-NMR spectrum of hybrid TF19 recorded on a Bruker Avance spectrometer (300 MHz, CDCl3):
0.0
S4
200 150 100 50
20.2 12.0
36.3 34.1 31.8 29.4 29.2 28.9 28.8 26.1 25.9 25.6 25.1 24.6 22.0 21.2
51.6 45.3 37.3
64.3
80.1
92.2 91.5
104.4
139.7
143.2
148.5 144.6
171.1
172.1
187.9 187.3
13
C-NMR spectrum of hybrid TF19 recorded on a Bruker Avance spectrometer (75 MHz, CDCl3):
0
S5
1.0
0.78
0.88
1.10 1.08 0.93 0.92
1.15
1.52 1.37 1.36
1.79
2.00 1.91
2.11
2.43
2.52 2.49 2.46
2.71
4.10 4.08 4.06 3.09 3.07 3.04 3.02 3.00 2.98 2.96
4.16
4.26
4.39
2.37 2.21
6.96 8.18 5.61
21.58
2.0
12.59
3.0 2.61 5.58 1.18 2.39
4.0 2.20 6.25
5.0 0.89
6.0 1.89 3.14 1.10
2.00
0.81
7.0 5.28
6.46 6.46
1
H-NMR spectrum of hybrid TF26 recorded on a Bruker Avance spectrometer (300 MHz, CDCl3):
0.0
S6
150 100 50
11.7
13.1 12.7
20.2 20.1
21.4
25.1 24.9 24.8
26.7 26.3 26.1 26.0
29.2 28.8
30.8 30.6 30.1
34.6 34.5 34.2
36.7
52.5 52.2 44.5 44.2 37.4
71.4 67.1 64.2
73.3
81.2 81.1
89.4 88.9
103.1 102.9
130.1
140.3
144.6
154.6
172.3 172.1
188.3 186.9
13
C-NMR spectrum of hybrid TF26 recorded on a Bruker Avance spectrometer (75 MHz, CDCl3):
0
S7
1
H-NMR spectrum of hybrid REI 213 recorded on a Bruker Avance spectrometer (300 MHz, CDCl3):
S8
13
C-NMR spectrum of hybrid REI 213 recorded on a Bruker Avance spectrometer (75 MHz, CDCl3):
S9
1
H-NMR spectrum of hybrid REI 220+216 recorded on a Bruker Avance spectrometer (300 MHz, CDCl3):
S10
13
C-NMR spectrum of hybrid REI 220+216 recorded on a Bruker Avance spectrometer (75 MHz, CDCl3):
S11
1
H-NMR spectrum of hybrid REI 234 recorded on a Bruker Avance spectrometer (300 MHz, CDCl3):
S12
13
C-NMR spectrum of hybrid REI 234 recorded on a JEOL JNM GX 400 spectrometer (100 MHz, CDCl3):
S13
1
H-NMR spectrum of hybrid REI 259 recorded on a JEOL JNM GX 400 spectrometer (400 MHz, Acetone-d6):
S14
13
C-NMR spectrum of hybrid REI 259 recorded on a JEOL JNM GX 400 spectrometer (100 MHz, Acetone-d6):
S15
1
H-NMR spectrum of hybrid HH3 recorded on a Bruker Avance spectrometer (300 MHz, Acetone-d6):
S16
13
C-NMR spectrum of hybrid HH3 recorded on a Bruker Avance spectrometer (75 MHz, Acetone-d6):
S17
1
H-NMR spectrum of hybrid VK3 recorded on a JEOL JNM GX 400 spectrometer (400 MHz, Acetone-d6):
S18
13
C-NMR spectrum of hybrid VK3 recorded on a JEOL JNM GX 400 spectrometer (100 MHz, Acetone-d6):
S19
Purple Dark green Orange Light blue Rosy Dark blue Grey Middle blue Dark brown Pink Light green Yellow Cyan blue Red Brown
Doxorubicin alone DOX + TF26 DOX + REI213 DOX + REI220+26 DOX + VK3 DOX + TF27 DOX + REI230 DOX + HH3 DOX + REI234 DOX + DHA DOX + TF29 DOX + TF19 DOX + REI259 DOX + REI235 DOX + Verapamil
Figure S2: Representative flow cytometric histograms of doxorubicin uptake in CEM/ADR5000 cells. Cells were incubated for 24 h with doxorubicin alone (purple curve on the left), doxorubicin (DOX) plus ART derivatives (from left to right: TF26, REI213, REI220+26, VK3, TF27, REI230, HH3, REI234, DHA, TF29, TF19, REI259, REI235), or doxorubicin plus verapamil (brown curve on the right. Shown are the fluorescence intensities (x-axis) and cell counts (y-axis).
S20
S21
