Red-Emitting Ruthenium(II) and Iridium(III) Complexes as. Phosphorescent Probes for Methylglyoxal in vitro and in vivo

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1 Supporting information for: Red-Emitting Ruthenium(II) and Iridium(III) Complexes as Phosphorescent Probes for Methylglyoxal in vitro and in vivo Wenzhu Zhang, *, Feiyue Zhang, Yong-Lei Wang, Bo Song, Run Zhang, *, Jingli Yuan State Key Laboratory of Fine Chemicals, School of Chemistry, Dalian University of Technology, Dalian , PR China (W. Zhang), Tel./Fax: Applied Physical Chemistry, Department of Chemistry, KTH Royal Institute of Technology, SE Stockholm, Sweden. Australian Institute for Bioengineering and anotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia (R. Zhang), Tel: , Fax:

2 General information for sample preparation Hydrogen peroxide (H 2 O 2 ) was diluted immediately from a stabilized 30% solution, and was assayed using its molar absorption coefficient of 43.6 M 1 cm 1 at 240 nm. 1, 2 Singlet oxygen ( 1 O 2 ) was generated from the a 2 MoO 4 -H 2 O 2 system in 0.05 M carbonate buffer of ph A stock solution of HOCl was prepared by dilution of the commercial sodium hypochlorite solution and stored according to the previous literatures. The concentration of HOCl was determined by using its molar extinction coefficient of 391 M 1 cm 1 at 292 nm before usage. 4 OOO - was donated by 3- morpholinosydnonimine. Hydroxylradical ( OH) was generated in the Fenton system from ferrous ammonium sulfate and hydrogen peroxide. itric oxide (O) aqueous solution was prepared by passing O gas through a deoxidized PBS buffer for 3 h, and the concentration was measured by using reported method. 5 Metal ions, including a +, K +, Mg 2+, Ca 2+, Zn 2+, Mg 2+, Fe 2+, Fe 3+, Cu 2+, and i 2+ were daily prepared as nitrate salts in DI water. Cell line and cell culture RAW macrophage cells were obtained from Dalian Medicine University. The cells were cultured in Dulbecco s modified Eagle s medium (DMEM, Sigma-Aldrich, Inc.), supplemented with 10% fetal bovine serum (FBS), 1% penicillin, 1% streptomycin sulphate in a humidified 5% CO 2 /95% air incubator at 37 o C. The growth medium was changed every two days. The cells were routinely subcultured using 0.05% trypsin-edta solution and growth to 80% confluence prior to experiments. Cytotoxicity analysis The cytotoxicity of the complexes towards RAW macrophage cells was examined by MTT analysis, which involves the reduction of [3-(4,5-dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide] tetrazolium to an insoluble formazan crystal by the metabolic activity of living cells. 6 RAW macrophage cells were seeded at a density of cells/ml in a 96-well micro-assay

3 culture plate and growth for 24 h at 37 o C in a 5% CO 2 /95% air incubator. Then, [Ru(bpy) 2 (DAphen)](PF 6 ) 2 or [Ir(ppy) 2 (DA-phen)](PF 6 ) in fresh culture medium was added into each well with different final concentrations of 0, 25, 50, 100, 150, 200 µm. Control wells were prepared by the addition of culture medium, and wells containing culture media without cells were used as blanks. After incubation at 37 o C in a 5% CO 2 /95% air incubator for 24 h, cell culture medium was removed and cells were washed three times with PBS. Then, 100 µl of 0.5 mg/ml MTT solution in PBS was added to each well, and the cells were incubated for another 4 h. The excess MTT solution was then carefully removed from each well, and the formed formazan was dissolved in 100 µl of DMSO (dimethyl sulfoxide). The optical density of each well was measured at a wavelength of 570 nm referenced at 690 nm using a microplate reader (Bio-Rad, xmark). The results from the five individual experiments were averaged. The following formula was used to calculate the viability of cell growth: Vialibity(%) = (mean of absorbance value of treatment group blank)/(mean absorbance value of control blank) 100. Ir Cl Cl Ir + H 2 H 2 (a)ch 2 Cl 2 /MeOH(1:2);reflux;4h (b)h 4 PF 6 ;1h Ir H 2 H 2 PF 6 Scheme S1. Synthetic pathway of [Ir(ppy) 2 (DA-phen)]PF 6.

4 Figure S1. 1 H MR spectrum of [Ir(ppy) 2 (DA-phen)]PF 6. Figure S2. 13 C MR spectrum of [Ir(ppy) 2 (DA-phen)]PF 6.

5 Figure S3. ESI-HRMS of [Ir(ppy) 2 (DA-phen)]PF 6. Figure S4. ITMS of [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 reacted with MGO.

6 Figure S5. ITMS of [Ir(ppy) 2 (DA-phen)](PF 6 ) reacted with MGO Current (µa) Potential (V) vs Ag/AgO 3 Figure S6. Cyclic voltammograms of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) (the product of [Ru(bpy) 2 (DA-phen)] 2+ reacted with MGO) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s.

7 Current (µa) Potential (V) vs Ag/AgO 3 Figure S7. Cyclic voltammograms of 1.0 mm [Ir(ppy) 2 (DA-phen)] + (dash line) and [Ir(ppy) 2 (MPphen)] + (solid line) (the product of [Ir(ppy) 2 (DA-phen)] + reacted with MGO) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s. 150 Current (µa) Potential (V) vs Ag/AgCl Figure S8. Cyclic voltammograms of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) (the product of [Ru(bpy) 2 (DA-phen)] 2+ reacted with MGO) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s.

8 Current (µa) Potential (V) vs Ag/AgCl Figure S9. Cyclic voltammograms of 1.0 mm [Ir(ppy) 2 (DA-phen)] + (dash line) and [Ir(ppy) 2 (MPphen)] + (solid line) (the product of [Ir(ppy) 2 (DA-phen)] + reacted with MGO) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s Current (µa) Potential (V) vs Ag/AgCl Figure S10. Differential pulse voltammetry of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s.

9 15 Current (µa) Potential (V) vs Ag/AgCl Figure S11. Differential pulse voltammetry of 1.0 mm [Ir(bpy) 2 (DA-phen)] + (dash line) and [Ir(bpy) 2 (MP-phen)] + (solid line) in 50 mm PBS buffer of ph 7.4. Scan rate: 0.1 V/s. 60 Current (µa) Potential (V) vs Ag/AgO 3 Figure S12. Differential pulse voltammetry of 1.0 mm [Ir(bpy) 2 (DA-phen)] + (dash line) and [Ir(bpy) 2 (MP-phen)] + (solid line) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s.

10 20 15 Current (µa) Potential (V) vs Ag/AgO 3 Figure S13. Differential pulse voltammetry of 1.0 mm [Ru(bpy) 2 (DA-phen)] 2+ (dash line) and [Ru(bpy) 2 (MP-phen)] 2+ (solid line) in CH 3 C containing 0.1 M TBAPF 6. Scan rate: 0.1 V/s ECL intensity (a.u.) Scan time (s) Figure S14. ECL behaviors of [Ru(bpy) 2 (DA-phen)] 2+ (100 µm) in the absence (dash line) and presence (solid line) of MGO (800 µm) in 50 mm PBS buffer of ph 7.4 containing 10 mm TPrA at room temperature. Scan rate of ECL was 0.15 v/s.

11 2000 ECL intensity (a.u.) Scan time (s) Figure S15. ECL behaviors of [Ir(bpy) 2 (DA-phen)] + (100 µm) in the absence (dash line) and presence (solid line) of MGO (800 µm) in 50 mm PBS buffer of ph 7.4 containing 10 mm TPrA at room temperature. Scan rate of ECL was 0.15 v/s. Luminescence intensity (a.u.) 250 A ph Luminescence intensity (a.u.) B ph Figure S16. Effects of ph on the phosphorescence responses of [Ru(bpy) 2 (DA-phen)] 2+ and [Ir(ppy) 2 (DA-phen)] + towards MGO. Phosphorescence intensities of [Ru(bpy) 2 (DA-phen)] 2+ (A, 10 µm in 25 mm PBS buffer) and [Ir(ppy) 2 (DA-phen)] + (B, 10 µm in 2:1 EtOH-25mM PBS buffer) at different ph values in the absence (, black line) and presence (, red line) of 800 µm MGO.

12 Intensity (a. u.) A a b c d PL Enhancement factor B a b c d e f Figure S17. (A) Phosphorescence intensity of (a) [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 (10 µm) in PBS buffer, (b) [Ru(bpy) 2 (MP-phen)](PF 6 ) 2 (10 µm) in 25 mm PBS buffer of ph 7.4, (c) [Ir(ppy) 2 (DAphen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v)and (d) [Ir(ppy) 2 (MP-phen)](PF 6 ) (10 µm) in PBS- EtOH (1:2, v/v) in the absence (grey bars) and presence (blank bars) of calf thymus DA; (B) changes in phosphorescence intensity of the probes and their reaction products with MGO in the absence (grey bars) and presence (black bars) of calf thymus DA. (a) [Ru(bpy) 2 (DA-phen)](PF 6 ) 2 (10 µm) in PBS buffer; (b) [Ru(bpy) 2 (MP-phen)](PF 6 ) 2 (10 µm) in 25 mm PBS buffer of ph 7.4; (c) [Ir(ppy) 2 (DA-phen)](PF 6 ) (10 µm) in PBS; (d) [Ir(ppy) 2 (MP-phen)](PF 6 ) (10 µm) in 25 mm PBS buffer of ph 7.4; (e) [Ir(ppy) 2 (DA-phen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v); (f) [Ir(ppy) 2 (MPphen)](PF 6 ) (10 µm) in PBS-EtOH (1:2, v/v).

13 Ru H 2 H 2 [Ru(bpy) 2 (DA-phen)] 2+ Figure S18. Optimized molecular geometry of [Ru(bpy) 2 (DA-phen)] 2+ in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S1. Cartesian coordinates of [Ru(bpy) 2 (DA-phen)] 2+ in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C C C C C C C C C C H H H H H H H H C C C C

14 23 C C C C C C H H H H H H H H C C C C C C C C C C H H H H H Ru C C H H H H H

15 Ru [Ru(bpy) 2 (DA-phen)] 2+ Figure S19. Optimized molecular geometry of [Ru(bpy) 2 (MP-phen)] 2+ in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S2. Cartesian coordinates of [Ru(bpy) 2 (MP-phen)] 2+ in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C C C C C C C C C C H H H H H H H H C C C C

16 23 C C C C C C H H H H H H H H C C C C C C C C C C H H H H Ru H C C H C C H H H C H

17 Ir H 2 H 2 [Ir(ppy) 2 (DA-phen)] + Figure S20. Optimized molecular geometry of [Ir(ppy) 2 (DA-phen)] + in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S3. Cartesian coordinates of [Ir(ppy) 2 (DA-phen)] + in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C C C C C C C C C C H H H H H H H H C

18 20 C C C C C C C C C H H H H H H H H C C C C C C C C C C H H H H H Ir C C H H H H H

19 Ir [Ir(ppy) 2 (MP-phen)] + Figure S21. Optimized molecular geometry of [Ir(ppy) 2 (MP-phen)] + in the ground state obtained from DFT calculations at B3LYP//6-311*G(d)//LAL2DZ level of theory. Table S4. Cartesian coordinates of [Ir(ppy) 2 (MP-phen)] + in the ground state. umber Atom Coordinates (ground state) X Y Z 1 C C C C C C C C C C H H H H H H H H C C C C

20 23 C C C C C C H H H H H H H H C C C C C C C C C C H H H H Ir H C C H C C H H H C H

21 Figure S22. TDDFT-based calculated absorption spectrum of [Ru(bpy) 2 (DA-phen)] 2+. Figure S23. TDDFT-based calculated absorption spectrum of [Ru(bpy) 2 (MP-phen)] 2+.

22 Figure S24. TDDFT-based calculated absorption spectrum of [Ir(ppy) 2 (DA-phen)] +. Figure S25. TDDFT-based calculated absorption spectrum of [Ir(ppy) 2 (MP-phen)] +.

23 Table S5. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ru(bpy) 2 (DA-phen)] 2+. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ru(bpy) 2 (D Electronic TDDFT// B3LYP//6-311*G(d) A-phen)] 2+ transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T HOMO LUMO 0.63 L LCT HOMO LUMO L LCT/L MCT S 0 T HOMO LUMO IL CT/L MCT S 0 T HOMO LUMO 0.66 L LCT HOMO LUMO L LCT/L MCT S 0 T HOMO LUMO 0.31 L LCT HOMO LUMO L LCT/L MCT S 0 T HOMO LUMO 0.26 L LCT HOMO LUMO L LCT/L MCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: bpy, L : DA-phen. Figure S26. Representative frontier molelcular orbital distributions of [Ru(bpy) 2 (DA-phen)] 2+ (isodensity contour = 0.02 a.u.).

24 Table S6. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ru(bpy) 2 (MP-phen)] 2+. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ru(bpy) 2 (M Electronic TDDFT// B3LYP//6-311*G(d) P-phen)] 2+ transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T HOMO-4 LUMO 0.14 L LCT/MLCT HOMO-3 LUMO 0.13 L LCT/MLCT HOMO LUMO 0.65 MLCT HOMO LUMO LL CT/ML CT S 0 T HOMO LUMO 0.69 MLCT S 0 T HOMO-1 LUMO 0.55 MLCT HOMO-2 LUMO 0.38 MLCT HOMO-1 LUMO MLCT/ML CT S 0 T HOMO-3 LUMO 0.10 LLCT/L LCT HOMO-2 LUMO 0.55 MLCT HOMO-1 LUMO 0.37 MLCT HOMO LUMO 0.11 MLCT S 0 T HOMO LUMO LL CT/ML CT HOMO LUMO MLCT HOMO LUMO ML CT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: bpy, L : MP-phen. Figure S27. Representative frontier molelcular orbital distributions of [Ru(bpy)2(MP-phen)]2+ (isodensity contour = 0.02 a.u.).

25 Table S7. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ir(ppy) 2 (DA-phen)] +. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ir(ppy) 2 (DAphen)] Electronic TDDFT// B3LYP//6-311*G(d) + transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T HOMO LUMO 0.24 L LCT/IL CT S 0 T HOMO LUMO L LCT/IL CT S 0 T HOMO LUMO L LCT S 0 T HOMO LUMO IL CT S 0 T HOMO LUMO L LCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: ppy, L : DA-phen. Figure S28. Representative frontier molelcular orbital distributions of [Ir(ppy) 2 (DA-phen)] + (isodensity contour = 0.02 a.u.).

26 Table S8. Selected Electronic Excitation Energies (ev) and corresponding Oscillator Strengths (f), main configurations and CI coefficients of the Low-lying Electronically Excited States of [Ir(ppy) 2 (MP-phen)] +. The calculation was conducted by TDDFT//B3LYP//6-311*G(d), based on the optimized molecular geometries in the ground state. [Ir(ppy) 2 (MPphen)] Electronic TDDFT// B3LYP//6-311*G(d) + transition Energy (ev) a f b Composition c CI d Assignment Triplet S 0 T HOMO LUMO 0.64 LL CT/LMCT HOMO LUMO L LCT/L MCT S 0 T HOMO LUMO 0.63 LL CT/LMCT HOMO LUMO L LCT/L MCT S 0 T HOMO LUMO LL CT HOMO LUMO LL CT S 0 T HOMO LUMO LL CT HOMO LUMO LL CT S 0 T HOMO LUMO L LCT a Only the selected low-lying excited state are presented. b Oscillator strength. c Only the main configuration are presented. d The CI coefficients are in absolute values. e L: ppy, L : MP-phen. Figure S29. Representative frontier molelcular orbital distributions of [Ir(ppy) 2 (MP-phen)] + (isodensity contour = 0.02 a.u.).

27 References 1. Miller, E. W.; Albers, A. E.; Pralle, A.; Isacoff, E. Y.; Chang, C. J., Boronate-Based Fluorescent Probes for Imaging Cellular Hydrogen Peroxide. J. Am. Chem. Soc.2005,127, Yuan, L.; Lin, W.; Xie, Y.; Chen, B.; Zhu, S., Single Fluorescent Probe Responds to H2O2, O, and H2O2/O with Three Different Sets of Fluorescence Signals. J. Am. Chem. Soc.2012,134, Ye, Z.; Song, B.; Yin, Y.; Zhang, R.; Yuan, J., Development of singlet oxygen-responsive phosphorescent ruthenium(ii) complexes. Dalton Trans.2013,42, Chen, X.; Wang, X.; Wang, S.; Shi, W.; Wang, K.; Ma, H., A Highly Selective and Sensitive Fluorescence Probe for the Hypochlorite Anion. Chem. Eur. J.2008,14, Zhang, R.; Ye, Z.; Wang, G.; Zhang, W.; Yuan, J., Development of a Ruthenium(II) Complex Based Luminescent Probe for Imaging itric Oxide Production in Living Cells. Chem. Eur. J.2010,16, Li, C.; Yu, M.; Sun, Y.; Wu, Y.; Huang, C.; Li, F., A onemissive Iridium(III) Complex That Specifically Lights-Up the uclei of Living Cells. J. Am. Chem. Soc.2011,133,