Journal: Biometals

Title: Biological and cytoselective anticancer properties of copper(II)-polypyridyl

complexes modulated by auxiliary methylated glycine ligand

Corresponding author: Chew-Hee Ng

(i)  Fig S1 Visible spectrum of 3

Fig S1 Visible spectrum of 1x10-3 M aqueous solution of 3

(ii)  Figs S 2.1 – 2.4 ESI-MS spectra of complexes 1-4

Fig S2.1 Positive-ion ESI mass spectrum of methanolic solution of 1, showing experimental isotopic distribution of [Cu(phen)(gly)]+ species and their theoretical calculated m/z

Fig S2.2 Positive-ion ESI mass spectrum of methanolic solution of 2, showing experimental isotopic distribution of [Cu(phen)(DL-ala)]+ species and their theoretical calculated m/z

Fig S2.3 Positive-ion ESI mass spectrum of methanolic solution of 3, showing experimental isotopic distribution of [Cu(phen)(C-dmg)]+ species and their theoretical calculated m/z

Fig S2.4 Positive-ion ESI mass spectrum of methanolic solution of 3, showing experimental isotopic distribution of [Cu(phen)(sar)]+ species and their theoretical calculated m/z

(iii)  Figs S 3.1 – 3.4 EPR spectra of complexes 1-4

Fig S3.1 EPR spectra of 1 on B-form DNA fibers at room temperature

Fig S3.2 EPR spectra of 2 on B-form DNA fibers at room temperature

Fig S3.3 EPR spectra of 3 on B-form DNA fibers at room temperature

Fig S3.4 EPR spectra of 4 on B-form DNA fibers at room temperature

(iv)  Figs S4.1 – S4.4 Gel electrophoresis results of restriction enzyme inhibition by complexes 1-4

Fig S4.1 Electrophoresis results of incubating λ DNA (0.5 μg/μL) with 5 unit of restriction enzyme in the absence (-) or presence (+) of 50 μM of complex 1 for 2 hours at 37ºC. Lane 1, 1 kb λ DNA; 2, λ DNA alone; 3, λ DNA + complex 1 alone. Restriction enzymes, Lanes 4 – 15: 4, Tsp 509I; 5, Hae III; 6, Sal I; 7, Pst I; 8, Pvu II; 9, Sca I; 10, Ssp I; 11, Ase I; 12, Mun I; 13, EcoR I; 14, NdeI; 15, Bst 11071.

Fig S4.2 Electrophoresis results of incubating λ DNA (0.5 μg/μL) with 5 unit of restriction enzyme in the absence (-) or presence (+) of 50 μM of complex 2 for 2 hours at 37ºC. Lane 1, 1 kb λ DNA; 2, λ DNA alone; 3, λ DNA + complex 2 alone. Restriction enzymes, Lanes 4 – 15: 4, Tsp 509I; 5, Hae III; 6, Sal I; 7, Pst I; 8, Pvu II; 9, Sca I; 10, Ssp I; 11, Ase I; 12, Mun I; 13, EcoR I; 14, NdeI; 15, Bst 11071.

Fig S4.3 Electrophoresis results of incubating λ DNA (0.5 μg/μL) with 5 unit of restriction enzyme in the absence (-) or presence (+) of 50 μM of complex 3 for 2 hours at 37ºC. Lane 1, 1 kb λ DNA; 2, λ DNA alone; 3, λ DNA + complex 3 alone. Restriction enzymes, Lanes 4 – 15: 4, Tsp 509I; 5, Hae III; 6, Sal I; 7, Pst I; 8, Pvu II; 9, Sca I; 10, Ssp I; 11, Ase I; 12, Mun I; 13, EcoR I; 14, NdeI; 15, Bst 11071.

Fig S4.4 Electrophoresis results of incubating λ DNA (0.5 μg/μL) with 5 unit of restriction enzyme in the absence (-) or presence (+) of 50 μM of complex 4 for 2 hours at 37ºC. Lane 1, 1 kb λ DNA; 2, λ DNA alone; 3, λ DNA + complex 4 alone. Restriction enzymes, Lanes 4 – 15: 4, Tsp 509I; 5, Hae III; 6, Sal I; 7, Pst I; 8, Pvu II; 9, Sca I; 10, Ssp I; 11, Ase I; 12, Mun I; 13, EcoR I; 14, NdeI; 15, Bst 11071.

Fig S5 Dose-response curves (a – e) of complexes 1 – 4 and [Cu(8OHQ)2] respectively, in nasopharyngeal cancer cell line HK1 and normal nasopharyngeal cell line NP69. Cells were incubated with increasing concentration of drugs for 24 h. Red curve () is for HK1 and blue curve () is for NP69.

Table S1 The CT-DNA apparent binding constants of complexes 1-4 in Tris-NaCl buffer at various concentration of NaCl (0-40 mM)

Complex / Buffer concentration (mM)* / Apparent binding constants
1 / T=5, N=0 / 5.12 ± 0.12 x 104
T=5, N=5 / 4.25 ± 0.09 x 104
T=5, N=10 / 3.33 ± 0.13 x 104
T=5, N=20 / 2.03 ± 0.14 x 104
T=5, N=40 / 1.56 ± 0.10 x 104
2 / T=5, N=0 / 5.88 ± 0.11 x 104
T=5, N=5 / 4.69 ± 0.10 x 104
T=5, N=10 / 3.96 ± 0.12 x 104
T=5, N=20 / 2.74 ± 0.13 x 104
T=5, N=40 / 1.95 ± 0.15 x 104
3 / T=5, N=0 / 5.54 ± 0.08 x 104
T=5, N=5 / 4.39 ± 0.15 x 104
T=5, N=10 / 3.72 ± 0.09 x 104
T=5, N=20 / 2.31 ± 0.12 x 104
T=5, N=40 / 1.68 ± 0.13 x 104
4 / T=5, N=0 / 3.65 ± 0.09 x 104
T=5, N=5 / 2.21 ± 0.15 x 104
T=5, N=10 / 1.35 ± 0.10 x 104
T=5, N=20 / 9.02 ± 0.16 x 103
T=5, N=40 / 6.31 ± 0.12 x 103
*T = Tris; N = NaCl