This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
1
SUPPORTING INFORMATION
Corroles and Corrole/Transferrin Nanoconjugates as Candidates for Sonodynamic Therapy
Vinay Kumar Sharma,a Atif Mahammed,a Matan Soll, a Boris Tumanskiia and Zeev
Grossa,*
aSchulich Faculty of Chemistry, Technion-Israel Institute of Technology, Haifa 32000, Israel.
samples with a known sample as standard. TMPone nitroxide was used as the standard sample,
in 10-5 M and 10-6 M concentration in our work (Fig. S6) for the technique called relative
measurement. So, the concentration of singlet oxygen produced by the sonosensitizers were
calculated by using the formula given below. The peak to peak amplitude were calculated as
shown in the Fig S7. One important note to bare in mind during this comparison that the EPR
signals must have the same linewidth. According to the formula, the concentration of singlet
oxygen produced by sonosensitizers are shown in the main text Fig 4. 1-H3 + Transferrin shows
highest intense signal which correspond to the amount of singlet oxygen formed in the system.
Fig S6: EPR signals of TMPone nitroxide as standard with concentration of (a) 10-6 M and (b)
10-5 M in PBS solution. Ultrasonic parameters were: US power, 3 W/cm2; time, 5 min; wave
frequency, 1 MHz.
10
Fig S7: Peak to peak amplitude and linewidth of an EPR signal.
UV-vis spectra:
Fig S8: Change in the UV-vis spectra of 1-H3 upon addition of 20 % apo-transferrin protein.
The spectra were recorded in PBS buffer solution.
11
0
0.05
0.1
0.15
0.2
0.25
350 450 550 650
Abs
orba
nce
Wavelength (nm)
1-Al1-Al+apo-Transferrin
Fig S9: Changes in the UV-vis spectra of 1-Al upon addition of 20 % apo-transferrin protein.
The spectra were recorded in PBS buffer solution.
0
0.1
0.2
350 400 450 500 550 600 650
Abs
orba
nce
Wavelength (nm)
1-Ga1-Ga+apo-Transferrin
Fig S10: Changes in the UV-vis spectra of 1-Ga upon addition of 20 % apo-transferrin protein.
The spectra were recorded in PBS buffer solution.
Cell proliferation assay and sonodynamic treatments:
12
Tumor cells DU-145 were seeded in 96-well microtiter plates (8×103 cells per well; 100 μL per
well) 24 h before the addition of the 2-Ga NPs. At the time of drug treatment, stock solutions
of compounds were diluted 10-fold to the desired final concentrations with EMEM medium.
Aliquots of 10 μL of these diluted solutions were added to the appropriate microtiter wells
containing 90 μL of growth medium, resulting in the required final drug concentrations (5
concentrations per compound, ranging from 0.02 to 27 μM). All cells were incubated in the
dark throughout the 24-72 h treatment period and did not receive prolonged exposure to light.
Following 24 h incubation at 37 °C, cells were exposed to ultrasound waves using a Saniflex
transducer (Chattanooga, Guildford Surrey, UK) at 1 MHz, 0.3 w/cm2 50% for 2 min. 24 h post
exposure to ultrasound cells viability was determined using the MTT assay (Sigma Aldrich)
according to the manufacturer’s instructions. Absorbance was measured using a microplate
reader (Synergy 4; Biotek Instruments) at 570 nm/630 nm. Experiments were performed in
triplicates and each dose response represents the mean of three or more independent
experiments.
0102030405060708090
0.02 5 9 17 27 37
% o
f cel
l sur
viva
l
2-H3 NPs (µM)
* * **
Fig S11: Effects of sonodynamic treatment on DU-145 Tumor cells growth. DU-145 cells were
incubated for 24 h with the 2-H3 NPs (at various concentration ranging from 0.02 to 37 μM)
and then exposed to very low intensity US (0.3 W/cm2 for 2 min at 1 MHz. Cell proliferation
was evaluated after 24, using the MTT assay. Data points are reported as mean ± SEM *P<0.01
vs. reciprocal treatment without US. Representative of three independent results.
13
0
20
40
60
80
100
120
2 9 15 27 57 107
% o
f sur
viva
l
1-Ga (µM)
* * *
Fig S12: Effects of sonodynamic treatment on DU-145 Tumor cells growth. DU-145 cells were
incubated for 24 h with the 1-Ga (at various concentration ranging from 2 to 107 μM) and then
exposed to very low intensity US (0.3 W/cm2 for 2 min at 1 MHz. Cell proliferation was
evaluated after 24, using the MTT assay. Data points are reported as mean ± SEM *P<0.05 vs.
reciprocal treatment without US. Representative of three independent results.
0102030405060708090
100
9 15 27 57 107
% o
f cel
l sur
viva
l
1-Al (µM)
Fig S13: Effects of sonodynamic treatment on DU-145 Tumor cells growth. DU-145 cells were
incubated for 24 h with the 1-Al (at various concentration ranging from 9 to 107 μM) and then
exposed to very low intensity US (0.3 W/cm2 for 2 min at 1 MHz. Cell proliferation was
evaluated after 24, using the MTT assay.
14
References:
1. A. Mahammed, I. Goldberg and Z. Gross, Org. Lett., 2001, 3, 3443-3446.2. A. Kanamori, M.-M. Catrinescu, A. Mahammed, Z. Gross and L. A. Levin, J. Neurochem., 2010,
114, 488-498.3. M. Soll, O. Bar Am, A. Mahammed, I. Saltsman, S. Mandel, M. B. H. Youdim and Z. Gross, ACS
Chem. Neurosci., 2016, 7, 1374-1382.4. Z. Gross, N. Galili and I. Saltsman, Angew. Chem. Int. Ed., 1999, 38, 1427-1429.5. J. Bendix, I. J. Dmochowski, H. B. Gray, A. Mahammed, L. Simkhovich and Z. Gross, Angew.
Chem. Int. Ed., 2000, 39, 4048-4051.6. A. Mahammed and Z. Gross, J. Inorg. Biochem., 2002, 88, 305-309.