Supporting Information

Transcription

1 Supporting Information Introducing the 4-Phenyl-1,2,3-Triazole Moiety as a Versatile Scaffold for the Development of Cytotoxic Ruthenium(II) and Osmium(II) Arene cyclometalates Christoph A. Riedl,, Lea S. Flocke, Michaela Hejl, Alexander Roller, Matthias H. M. Klose,, Michael A. Jakupec,, Wolfgang Kandioller,,* and Bernhard K. Keppler, Institute of Inorganic Chemistry, Faculty of Chemistry, and Research Platform Translational Cancer Therapy Research - Waehringer Str. 42, 1090 Vienna, Austria * wolfgang.kandioller@univie.ac.at, Tel: ; Fax: Synthesis of triazole ligands (1a e) Single crystal X-ray diffraction analysis Stability in aqueous solution by UV/Vis spectroscopy Stability toward aqueous phosphate buffer by NMR spectroscopy Stability and amino acid interaction via ESI-MS Aquation-anation equilibria Topoisomerase IIα inhibition Cell cycle distribution H and 13 C NMR spectra

2 1. Synthesis of triazole ligands (1a e) General procedure. The respective organic halide (1.0 eq) was added to a solution of sodium azide (1.1 eq) in anhydrous DMSO (2 ml per mmol halide) and stirred at room temperature under argon atmosphere for 3 to 24 h. Deionized water (2 ml per mmol halide) was added, followed by sodium ascorbate (0.2 eq), phenylacetylene (1.0 eq) and aqueous copper sulfate solution (1 M, 0.2 eq). The reaction mixture was stirred vigorously under inert conditions for 8 to 48 h, after which the product was precipitated with deionized water and stored at 4 C overnight to complete precipitation. The formed solid was collected by filtration, washed with water and purified by column chromatography on silica to afford the desired triazole ligands 1a f. 1-Benzyl-4-phenyl-1,2,3-triazole (1a). The reaction was performed according to the general procedure for the synthesis of 1,2,3-triazoles, using benzyl bromide (1.42 ml, 12.0 mmol, 1.0 eq), sodium azide (859 mg, 13.2 mmol, 1.0 eq), phenylacetylene (1.32 ml, 12.0 mmol. 1.0 eq), sodium ascorbate (476 mg, 2.4 mmol, 0.2 eq) and copper sulfate pentahydrate (600 mg, 2.4 mmol, 0.2 eq) with a reaction time of 3 h for the conversion to the azide, 24 h for the cycloaddition step and subsequent purification on silica (10% acetone / chloroform). Yield: mg, 93%. ESI-HR-MS + m/z Found (Calculated): [M+H] ( ), [M+Na] ( ), [2M+Na] ( ). Elemental Anal. Calc. for C 15 H 13 N 3 : C 76.57, H 5.57, N 17.86; Found: C 76.32, H 5.20, N H NMR (DMSO-d 6, MHz, K): δ = 8.63 (s, 1H, ArH Trz-5 ); (m, 2H, ArH Ph-2 /6 ); (m, 2H, ArH Ph-3 /5 ); (m, 6H, ArH Ph-4, ArH Bn-2 /6, ArH Bn-3 /5, ArH Bn-4 ); 5.65 (s, 2H, CH 2, Bn ) ppm. 13 C NMR (DMSO-d 6, MHz, K): δ = (C Trz-4 ); (C Bn-1 ); (C Ph-1 ); (CH Ph-3 /5, CH Bn-3 /5, CH Bn-4 ); (CH Ph-4 ); (CH Bn-2 /6 ); (CH Ph-2 /6 ); (CH Trz-5 ); (CH 2, Bn ) ppm. 1-(4 -Methoxybenzyl)-4-phenyl-1,2,3-triazole (1b). The reaction was performed according to the general procedure for the synthesis of 1,2,3-triazoles, using 4-methoxybenzyl chloride (0.81 ml, 6.0 mmol, 1.0 eq), sodium azide (390 mg, 6.6 mmol, 1.1 eq), phenylacetylene (0.66 ml, 6.0 mmol, 1.0 eq), sodium ascorbate (240 mg, 1.2 mmol, 0.2 eq) and copper sulfate pentahydrate (300 mg, 1.2 mmol, 0.2 eq) with a reaction time of 16 h for the conversion to the azide, 8 h for the cycloaddition step and subsequent purification on silica (5% acetone / chloroform). Yield: 864 mg, 54%. ESI-HR-MS + m/z Found (Calculated): [M+H] ( ), [M+Na] ( ), [2M+Na] ( ). Elemental Anal. Calc. for C 16 H 15 N 3 O 0.1H 2 O: C 71.94, H 5.74, N 15.73, S 0.00; Found: C 72.12, H 5.68, N 15.46, S < H NMR (DMSO-d 6, MHz, K): δ = 8.58 (s, 1H, ArH Trz-5 ); (m, 2H, ArH Ph-2 /6 ); 7.43 (dd, 3 J H,H = 8 Hz, 8 Hz, 2H, ArH Ph-3 /5 ); (m, 3H, ArH Ph-4, ArH Bn-2 /6 ); (m, 2H, ArH Bn-3 /5 ); 5.56 (s, 2H, CH 2, Bn ) ppm. 13 C NMR (DMSO-d 6, MHz, K): δ = (C Bn-4 ); (C Trz-4 ); (C Ph-1 ); (CH Bn-2 /6 ); (CH Ph-3 /5 ); (C Bn-1 ); (CH Ph-4 ); (CH Ph-2 /6 ); (CH Trz-5 ); (CH Bn-3 /5 ); 55.1 (OCH 3, Bn ); 52.6 (CH 2, Bn ) ppm. 2

3 1-Propyl-4-phenyl-1,2,3-triazole (1c). The reaction was performed according to the general procedure for the synthesis of 1,2,3-triazoles, using 1-bromopropane (2.18 ml, 24.0 mmol, 1.0 eq), sodium azide (1.716 mg, 26.4 mmol, 1.1 eq), phenylacetylene (2.64 ml, 24.0 mmol, 1.0 eq), sodium ascorbate (950 mg, 4.8 mmol, 0.2 eq) and copper sulfate pentahydrate (1200 mg, 4.8 mmol, 0.2 eq) with a reaction time of 24 h for the conversion to the azide, 48 h for the cycloaddition step and subsequent purification on silica (5% acetone / chloroform). Yield: 2408 mg, 54%. ESI-HR-MS + m/z Found (Calculated): [M+H] ( ), [M+Na] ( ), [2M+Na] ( ). Elemental Anal. Calc. for C 11 H 13 N 3 : C 70.56, H 7.00, N 22.44; Found: C 70.48, H 6.92, N H NMR (DMSO-d 6, MHz, K): δ = 8.57 (s, 1H, ArH Trz-5 ); (m, 2H, ArH Ph-2 /6 ); (m, 2H, ArH Ph-3 /5 ); (m, 1H, ArH Ph-4 ); 4.36 (t, 3 J H,H = 7 Hz, 2H, CH 2, Prop-1 ); 1.89 (tq, 3 J H,H = 7 Hz, 7 Hz, 2H, CH 2, Prop-2 ); 0.88 (t, 3 J H,H = 7 Hz, 3H, CH 3, Prop-3 ) ppm. 13 C NMR (DMSO-d 6, MHz, K): δ = (C Trz-4 ); (C Ph-1 ); (CH Ph-3 /5 ); (CH Ph-4 ); (CH Ph-2 /6 ); (CH Trz-5 ); 51.1 (CH 2, Prop ); 23.1 (CH 2, Prop ); 10.8 (CH 3, Prop ) ppm. 1-Butyl-4-phenyl-1,2,3-triazole (1d). The reaction was performed according to the general procedure for the synthesis of 1,2,3-triazoles, using 1-bromobutane (0.32 ml, 3.0 mmol, 1.0 eq), sodium azide (214 mg, 3.3 mmol, 1.1 eq), phenylacetylene (0.33 ml, 3.0 mmol, 1.0 eq), sodium ascorbate (120 mg, 0.6 mmol, 0.2 eq) and copper sulfate pentahydrate (150 mg, 0.6 mmol, 0.2 eq) with a reaction time of 24 h for the conversion to the azide, 24 h for the cycloaddition step and subsequent purification on silica (10% acetone / chloroform). Yield: 413 mg, 68%. ESI-HR-MS + m/z Found (Calculated): [M+H] ( ), [M+Na] ( ), [2M+Na] ( ). Elemental Anal. Calc. for C 12 H 15 N 3 : C 71.61, H 7.51, N 20.88; Found: C 71.69, H 7.51, N H NMR (DMSO-d 6, MHz, K): δ = 8.57 (s, 1H, ArH Trz-5 ); (m, 2H, ArH Ph-2 /6 ); 7.44 (dd, 3 J H,H = 8 Hz, 8 Hz, 2H, ArH Ph-3 /5 ); 7.32 (dd, 3 J H,H = 8 Hz, 8 Hz, 1H, ArH Ph-4 ); 4.39 (t, 3 J H,H = 7 Hz, 2H, CH 2, But ); 1.84 (tt, 3 J H,H = 7 Hz, 7 Hz, 2H, CH 2, But ); 1.28 (tq, 3 J H,H = 7 Hz, 7 Hz, 2H, CH 2, But ); 0.90 (t, 3 J H,H = 7 Hz, 3H, CH 3, But ) ppm. 13 C NMR (DMSO-d 6, MHz, K): δ = (C Trz-4 ); (C Ph-1 ); (CH Ph-3 /5 ); (CH Ph-4 ); (CH Ph-2 /6 ); (CH Trz-5 ); 49.2 (CH 2, But ); 31.6(CH 2, But ); 19.1(CH 2, But ); 13.3 (CH 3, But ) ppm. Methyl 2 -(4-phenyl-1,2,3-triazol-1-yl)acetate (1e). The reaction was performed according to the general procedure for the synthesis of 1,2,3-triazoles, using methyl bromoacetate (1.14 ml, 12.0 mmol, 1.0 eq), sodium azide (860 mg, 13.2 mmol, 1.1 eq), phenylacetylene (1.32 ml, 12.0 mmol, 1.0 eq), sodium ascorbate (480 mg, 2.4 mmol, 0.2 eq) and copper sulfate pentahydrate (600 mg, 2.4 mmol, 0.2 eq) with a reaction time of 24 h for the conversion to the azide, 72 h for the cycloaddition step and subsequent purification on silica (5% acetone / chloroform). Yield: 1750 mg, 67%. ESI-HR-MS + m/z Found (Calculated): [M+H] ( ), [M+Na] ( ), [2M+Na] ( ). Elemental Anal. Calc. for C 11 H 11 N 3 O 2 : C 60.82,H 5.10, N 19.34; found: C 60.72, H 4.89, N H NMR (DMSO-d 6, MHz, K): δ = 8.56 (s, 1H, ArH Trz-5 ); 7.86 (d, 3 J H,H = 8 Hz, 2H, ArH Ph-2 /6 ); 7.46 (dd, 3 J H,H = 8 Hz, 8 Hz, 2H, ArH Ph-3 /5 ); 7.35 (dd, 3 J H,H = 8 Hz, 8 Hz, 1H, ArH Ph-4 ); 5.48 (s, 2H, CH 2, Ac ); 3.74 (s, 3H, OCH 3, Ac ) ppm. 13 C NMR (DMSO-d 6, MHz, K): δ = (CO Ac ), (C Trz-4 ); (C Ph-1 ); (CH Ph-3 /5 ); (CH Ph-4 ); (CH Ph-2 /6 ); (CH Trz-5 ); 52.6 (CH 3, Ac ); 50.5 (CH 2, Ac ) ppm. 3

4 2. Single crystal X-ray diffraction analysis Sample Diffractometer Source Table S1. Experimental parameters and CCDC codes. Temp. [K] Detector distance [mm] Time per frame [s] No. of frames Frame width [ ] CCDC 2a X8 Mo b D8 Mo c X8 Mo d D8 Mo e D8 Mo a X8 Mo b X8 Mo c X8 Mo d X8 Mo e D8 Mo Table S2. Half sandwich geometry parameter summary. Compound 2a 2b 2c 2d Metal Ru Ru Ru Ru Hapticity η π-plane centroid distance (dpi) [Å] 1.696(3) 1.699(3) (4) (4) (8) (7) Ligand plane centroid distance (dl) [Å] 1.388(3) (4) (3) (6) (5) Ring-slippage a [Å] (3) π-plane to ligand plane angle [ ] 5.0(2) 6.06(19) (3) (2) (19) (15) Angle (π-plane M Cl)[ ] Angle (π-plane M N3) [ ] Angle (π-plane M C2 ) [ ] Compound 2e 3a 3b 3c 3d 3e Metal Ru Os Os Os Os Os Hapticity η π-plane centroid distance (dpi) [Å] (4) (7) (7) (9) (9) 1.699(3) Ligand plane centroid distance (dl) [Å] (3) (6) (6) (7) (8) 1.399(3) Ring-slippage a [Å] π-plane to ligand plane angle [ ] (16) (15) (3) (19) (2) 6.06(19) Angle (π-plane M Cl) [ ] Angle (π-plane M N3) [ ] Angle (π-plane M C2 ) [ ] a Distance between the perpendicular projection of an heavy atom on the ring l.s.-plane and the arene ring 4

5 Figure S1. π-plane centroid distance (dpi) and ligand plane centroid distance (dl). Figure S2. Exemplary lattice of 3c (left - orthogonal layers) and 2e (right - parallel layers) to illustrate the two different crystallization patterns that dimeric interaction pairs can adopt. 5

6 Figure S3. Schematic representation of proposed π- π and π-σ interactions in 3c, representative for 2b e & 3a e (1: π Ph π Trz ; 2: π Trz π Trz ; 3: π Trz σ TrzH ; 4: π Ph σ TrzH ; 5,6: π Ph σ CH2 ). Figure S4. Schematic representation of alternating π trz -π Ph interactions found exclusively in crystals of 2a, in contrast to the other metalacycles represented in Figure S2. 6

7 Table S3. Selected bond lengths. Compound Bond lengths [Å] M Cl M N3 M C C1 -C2 C2 -C3 C3 -C4 C4 -C5 C5 -C6 C6 -C1 2a A (18) 2.069(6) 2.057(6) 1.429(9) 1.431(9) 1.387(9) 1.394(10) 1.401(10) 1.383(9) 2a B (17) 2.067(6) 2.063(6) 1.418(9) 1.416(9) 1.365(8) 1.410(10) 1.383(9) 1.398(9) 2b A (7) 2.072(2) 2.063(2) 1.410(3) 1.401(4) 1.394(4) 1.389(4) 1.388(4) 1.400(3) 2b B (7) 2.066(2) 2.072(2) 1.416(3) 1.399(4) 1.396(4) 1.390(4) 1.381(4) 1.393(3) 2c (12) 2.073(3) 2.065(4) 1.382(7) 1.378(6) 1.402(6) 1.407(6) 1.388(6) 1.392(6) 2d (3) (10) (12) (17) (17) (19) (17) (19) 1.388(2) 2e (3) (10) (12) (16) (17) (18) (17) (16) (16) 3a (15) 2.071(6) 2.089(6) 1.396(10) 1.404(9) 1.408(9) 1.365(11) 1.378(11) 1.408(8) 3b (13) 2.080(4) 2.096(5) 1.414(7) 1.408(7) 1.396(7) 1.370(8) 1.380(7) 1.387(6) 3c (14) 2.080(4) 2.095(5) 1.385(8) 1.364(8) 1.429(7) 1.382(8) 1.390(7) 1.392(7) 3d (12) 2.088(4) 2.079(4) 1.383(7) 1.420(6) 1.401(7) 1.392(7) 1.363(7) 1.389(6) 3e (8) 2.075(3) 2.084(3) 1.418(5) 1.401(5) 1.388(5) 1.391(6) 1.383(5) 1.397(5) 1a b Table S4. Selected bond lengths. Compound Bond lengths [Å] N1 N2 N2 N3 N3 C4 C4 C5 C5 N1 C4 C1 N1 C1 2a A 1.338(7) 1.329(8) 1.367(8) 1.372(10) 1.365(8) 1.451(10) 1.451(8) 2a B 1.351(7) 1.324(7) 1.368(8) 1.382(9) 1.345(8) 1.472(9) 1.470(8) 2b A 1.343(3) 1.320(3) 1.362(3) 1.372(3) 1.350(3) 1.457(3) 1.466(3) 2b B 1.343(3) 1.313(3) 1.374(3) 1.371(3) 1.353(3) 1.453(3) 1.456(3) 2c 1.341(5) 1.328(5) 1.373(5) 1.354(6) 1.351(5) 1.445(6) 1.467(5) 2d (14) (14) (15) (16) (15) (16) (15) 2e (14) (14) (15) (16) (15) (16) (15) 3a 1.352(8) 1.315(7) 1.368(8) 1.364(10) 1.362(8) 1.446(10) 1.439(9) 3b 1.332(5) 1.326(5) 1.368(6) 1.363(7) 1.339(6) 1.452(7) 1.475(6) 3c 1.350(5) 1.328(6) 1.365(6) 1.359(8) 1.352(7) 1.441(8) 1.464(7) 3d 1.342(5) 1.328(5) 1.366(5) 1.361(6) 1.351(6) 1.448(6) 1.462(6) 3e 1.341(4) 1.321(4) 1.368(4) 1.368(5) 1.354(5) 1.454(5) disordered 1a b CCDC code: NILLEZ01 7

8 Table S5. Selected bond angles Compound Bond angles [ ] C2 M Cl Cl M N3 N3 M C2 M C2 C1 M C2 C3 C1 C4 C5 C1 C4 N3 M N3 N2 M N3 C4 2a A 87.29(17) 84.99(15) 77.4(3) 117.4(5) 128.1(5) 138.5(6) 115.5(6) 130.4(4) 117.5(5) 2a B 86.95(16) 86.58(14) 77.5(2) 117.0(5) 115.0(6) 138.9(6) 114.0(6) 131.4(4) 118.3(4) 2b A 84.99(7) 88.10(6) 77.17(9) (18) (19) 138.7(2) 114.5(2) (16) (16) 2b B 85.97(7) 85.30(6) 77.29(9) (18) (19) 139.6(2) 114.1(2) (16) (16) 2c 85.41(12) 88.73(10) 77.11(16) 116.9(3) 127.6(3) 139.0(4) 114.2(4) 131.6(3) 117.7(3) 2d 86.00(3) 88.85(3) 77.32(4) (8) (10) (11) (10) (8) (8) 2e 85.89(3) 89.25(3) 77.43(4) (8) (9) (11) (10) (8) (8) 3a 85.62(16) 85.44(13) 76.1(3) 117.2(5) 125.9(6) 138.9(7) 113.8(7) 129.9(4) 119.0(5) 3b 86.79(14) 85.84(12) 76.34(18) 117.1(4) 127.0(4) 139.1(4) 114.9(4) 130.3(3) 118.5(3) 3c 85.38(15) 87.27(13) 76.78(19) 115.9(4) 128.2(4) 138.4(5) 114.8(5) 130.4(3) 118.5(4) 3d 85.76(12) 87.03(10) 76.78(17) 116.7(3) 128.4(4) 139.1(4) 114.7(4) 130.5(3) 111.4(4) 3e Fehlt fehlt 76.80(12) 117.0(2) 127.9(3) 138.2(3) 115.0(3) 130.6(2) 118.2(2) 1a Table S6. Selected torsion angles. Torsion angles [ ] Compound C5 N1 C1 C1 N1 C5 C4 N3 C4 C1 C2 C2 2a A 177.2(5) -4.4(8) 82.7(8) 2a B (5) -2.9(7) 90.4(7) 2b A 178.6(2) 0.7(3) -80.4(3) 2b B 173.4(2) 2.8(3) -86.9(3) 2c (4) -0.6(6) 59.0(6) 2d (10) -0.37(14) 69.13(15) 2e (11) -1.81(15) 96.26(14) 3a (6) -2.5(8) 89.3(8) 3b (5) -2.7(6) 73.9(6) 3c (5) -1.5(7) 58.7(7) 3d (4) -0.6(6) 69.7(6) 3e disordered -0.7(4) disordered 8

9 Table S7. Selected π-π interaction parameters. Triazole ring Triazole ring Triazole ring Phenyl ring Compound Centroid-centroid distance [Å] Normal distance [Å] Displacement angle c [ ] Plane to plane angle [ ] Centroid-centroid distance [Å] Normal distance [Å] Displacement angle [ ] Plane to plane angle [ ] 2a A-B a B-A b A-A b B-B c d e a b c d e Table S8. Selected σ-π interaction parameters. Triazole C 5 H - Triazole ring Triazole C 5 H - Phenyl ring Compound Atom-centroid distance [Å] Normal distance [Å] Displacement angle [ ] Atom-centroid distance [Å] Normal distance [Å] Displacement angle [ ] 2a A-B a B-A b A-A b B-B c d e a b c d e c Centroid-centroid distances and ring normals were determined from experimental data by OLEX2, displacement angles were calculated parameters by use of the sin function and are given without standard deviation. 9

10 Table S9. Selected σ-π interaction parameters. CH 2, A Phenyl ring CH 2, B Phenyl ring Compound Atom-centroid distance [Å] Normal distance [Å] Displacement angle [ ] Atom-centroid distance [Å] Normal distance [Å] Displacement angle [ ] 2b A-A b B-B c d e a b c d e Disordered Disordered 10

11 2.1. [Chlorido(1-benzyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)ruthenium(ii)] (2a) Figure S5. Crystal structure of 2a, drawn with 50% displacement ellipsoids, second molecule of asymmetric unit omitted for clarity. Table S10. Sample and crystal data of 2a. Chemical formula C 25 H 26 ClN 3 Ru Crystal system triclinic Formula weight [g/mol] Space group P-1 Temperature [K] 140 Z 4 Measurement method \Φ and \ω scans Volume [Å 3 ] (12) Radiation wavelength [Å] MoK α (λ = ) (3) (16) Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (4) (15) Crystal habit clear orange needle (5) (15) Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T max Abs. correction type multi-scan F(000) [e - ]

12 Table S11. Data collection and structure refinement of 2a. Index ranges -12 h k l 22 Theta range for data collection [ ] to Reflections number Refinement method Least squares Function minimized Σ w(f o 2 - F c 2 ) 2 Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] Data / restraints / parameters Final R indices all data I>2σ(I) 8074 / 72 / 547 R 1 = wr 2 = R 1 = wr 2 = w=1/[σ 2 (F o 2 )+(0.0531P) 2 ] Weighting scheme 1.09 / where P=(F 2 o +2F 2 c )/3 12

13 2.2. [Chlorido(1-(4 -methoxybenzyl)-4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazole)(η 6 -p-cymene)ruthenium(ii)] (2b) Figure S6. Crystal structure of 2b, drawn with 50% displacement ellipsoids, second molecule of asymmetric unit omitted for clarity. Table S12. Sample and crystal data of 2b. Chemical formula C 26 H 28 ClN 3 ORu Crystal system triclinic Formula weight [g/mol] Space group P-1 Temperature [K] 100 Z 4 Measurement method \Φ and \ω scans Volume [Å 3 ] (13) Radiation wavelength [Å] MoK α (λ = ) Unit cell (4) (12) Crystal size [mm 3 ] dimensions [Å] and (4) (12) Crystal habit clear orange block [ ] (5) (10) Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T Abs. correction type multi-scan F(000) [e - ]

14 Table S13. Data collection and structure refinement of 2b. Index ranges -16 h k l 20 Theta range for data collection [ ] to Reflections number Data / restraints / parameters / 0 / 585 Refinement method Least squares all data all data Function minimized Σ w(f 2 o - F 2 c )2 Final R indices I>2σ(I) I>2σ(I) Goodness-of-fit on F w=1/[σ 2 (F 2 o )+(0.0284P) P] Largest diff. peak and hole [e Å -3 ] 0.58/-0.63 Weighting scheme where P=(F 2 o +2F 2 c )/3 14

15 2.3. [Chlorido(1-propyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)ruthenium(ii)] (2c) Figure S7. Asymmetric unit of 2c, drawn with 50% displacement ellipsoids. Table S14. Sample and crystal data of 2c. Chemical formula C 21 H 26 ClN 3 Ru Crystal system tetragonal Formula weight [g/mol] Space group I41/a Temperature [K] 120 Z 16 Measurement method \Φ and \ω scans Volume [Å 3 ] (10) Radiation wavelength [Å] MoK α (λ = ) (15) 90 Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (15) 90 Crystal habit clear yellow needle (6) 90 Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T Abs. correction type multi-scan F(000) [e - ]

16 Table S15. Data collection and structure refinement of 2c. Index ranges -33 h k l 12 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 3658 / 0 / 239 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F 2 1 w=1/[σ 2 (F 2 o )+(0.0179P) P] Largest diff. peak and hole [e Å -3 ] 0.44 / Weighting scheme where P=(F 2 o +2F 2 c )/3 16

17 2.4. [Chlorido(1-butyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)ruthenium(ii)] (2d) Figure S8. Asymmetric unit of 2d, drawn with 50% displacement ellipsoids. Table S16. Sample and crystal data of 2d. Chemical formula C 22 H 28 ClN 3 Ru Crystal system tetragonal Formula weight [g/mol] Space group I41/a Temperature [K] 100 Z 16 Measurement method \Φ and \ω scans Volume [Å 3 ] (8) Radiation wavelength [Å] MoK α (λ = ) (11) 90 Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (11) 90 Crystal habit clear orange block (5) 90 Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T Abs. correction type multi-scan F(000) [e - ]

18 Table S17. Data collection and structure refinement of 2d. Index ranges -40 h k l 14 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 6140 / 0 / 248 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F w=1/[σ 2 (F 2 o )+(0.0196P) P] Largest diff. peak and hole [e Å -3 ] 0.46 / Weighting scheme where P=(F 2 o +2F 2 c )/3 18

19 2.5. [Chlorido(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazol-1- yl)acetate)(η 6 -p-cymene)ruthenium(ii)] (2e) Figure S9. Asymmetric unit of 2e, drawn with 50% displacement ellipsoids. Table 18. Sample and crystal data of 2e. Chemical formula C 21 H 24 ClN 3 O 2 Ru Crystal system triclinic Formula weight [g/mol] Space group P-1 Temperature [K] 100 Z 2 Measurement method \Φ and \ω scans Volume [Å 3 ] (6) Radiation wavelength [Å] MoK α (λ = ) (3) (10) Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (3) (14) Crystal habit clear orange block (5) (10) Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T max Abs. correction type multi-scan F(000) [e - ]

20 Table S19. Data collection and structure refinement of 2e. Index ranges -13 h k l 16 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 6090 / 0 / 257 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0222P) P] Weighting scheme 0.52 / where P=(F 2 o +2F 2 c )/3 20

21 2.6. [Chlorido(1-benzyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)osmium(ii)] 3a Figure S10. Asymmetric unit of 3a, drawn with 50% displacement ellipsoids. Table S20. Sample and crystal data of 3a, solvent molecules omitted for clarity. Chemical formula C 26 H 27 Cl 4 N 3 Os Crystal system monoclinic Formula weight [g/mol] Space group P21/n Temperature [K] 130 Z 4 Measurement method \Φ and \ω scans Volume [Å 3 ] (3) Radiation wavelength [Å] MoK α (λ = ) (8) 90 Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (2) (2) Crystal habit clear yellow plate (6) 90 Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T max Abs. correction type multi-scan F(000) [e - ]

22 Table S21. Data collection and structure refinement of 3a. Index ranges -11 h k l 9 Theta range for data collection [ ] 3.15 to Reflections number Data / restraints / parameters 4842 / 1 / 301 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0239P) P] Weighting scheme 1.46 / where P=(F 2 o +2F 2 c )/3 22

23 2.7. [Chlorido(1-(4 -methoxybenzyl)-4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazole)(η 6 -p-cymene)osmium(ii)] (3b) Figure S11. Asymmetric unit of 3b, drawn with 50% displacement ellipsoids. Table S22. Sample and crystal data of 3b. Chemical formula C 26 H 28 ClN 3 OOs Crystal system triclinic Formula weight [g/mol] Space group P-1 Temperature [K] 140 Z 2 Measurement method \Φ and \ω scans Volume [Å 3 ] (13) Radiation wavelength [Å] MoK α (λ = ) (6) (3) Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (7) (3) Crystal habit clear yellow plate (8) (2) Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min 0.62 Abs. correction T max Abs. correction type multi-scan F(000) [e - ]

24 Table S23. Data collection and structure refinement of 3b. Index ranges -11 h k l 14 Theta range for data collection [ ] 4.19 to Reflections number Data / restraints / parameters 4170 / 0 / 293 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0250P) 2 ] Weighting scheme 0.83 / where P=(F 2 o +2F 2 c )/3 24

25 2.8. [Chlorido(1-propyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)osmium(ii)] (3c) Figure S12. Asymmetric unit of 3c, drawn with 50% displacement ellipsoids. Table S24. Sample and crystal data of 3c. Chemical formula C 21 H 26 ClN 3 Os Crystal system tetragonal Formula weight [g/mol] Space group I41/a Temperature [K] 120 Z 16 Measurement method \Φ and \ω scans Volume [Å 3 ] (11) Radiation wavelength [Å] MoK α (λ = ) (16) 90 Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (16) 90 Crystal habit clear yellow needle (7) 90 Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T Abs. correction type multi-scan F(000) [e - ]

26 Table S25. Data collection and structure refinement of 3c. Index ranges -21 h k l 8 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 3686 / 0 / 239 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0231) 2 ] Weighting scheme 0.72 / where P=(F 2 o +2F 2 c )/3 26

27 2.9. [Chlorido(1-butyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)osmium(ii)] (3d) Figure S13. Asymmetric unit of 3d, drawn with 50% displacement ellipsoids. Table S26. Sample and crystal data of 3d. Chemical formula C 22 H 28 ClN 3 Os Crystal system tetragonal Formula weight [g/mol] Space group I41/a Temperature [K] 120 Z 16 Measurement method \Φ and \ω scans Volume [Å 3 ] (12) Radiation wavelength [Å] MoK α (λ = ) (19) 90 Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (19) 90 Crystal habit clear yellow needle (7) 90 Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T Abs. correction type multi-scan F(000) [e - ]

28 Table S27. Data collection and structure refinement of 3d. Index ranges -34 h k l 12 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 3747 / 0 / 248 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0129P) 2 ] Weighting scheme 0.96 / where P=(F 2 o +2F 2 c )/3 28

29 2.10. [Chlorido(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazol-1- yl)acetate)(η 6 -p-cymene)osmium(ii)] (3e) Figure S14. Asymmetric unit of 3e, drawn with 50% displacement ellipsoids. Solvent molecules and disordered moieties omitted for clarity. Table S28. Sample and crystal data of 3e. Chemical formula C 21.5 H 25 Cl 2 N 3 O 2 Os Crystal system triclinic Formula weight [g/mol] Space group P-1 Temperature [K] 100 Z 1 Measurement method \Φ and \ω scans Volume [Å 3 ] (6) Radiation wavelength [Å] MoK α (λ = ) (3) (13) Crystal size [mm 3 ] Unit cell dimensions [Å] and [ ] (3) (12) Crystal habit clear yellow block (3) (12) Density (calculated) [g/cm 3 ] Absorption coefficient [mm -1 ] Abs. correction T min Abs. correction T max Abs. correction type multi-scan F(000) [e - ]

30 Table S29. Data collection and structure refinement of 3e. Index ranges -12 h k l 13 Theta range for data collection [ ] to Reflections number Data / restraints / parameters 4118 / 2 / 325 Refinement method Least squares all data all data Final R indices Function minimized Σ w(f 2 o - F 2 c ) 2 I>2σ(I) I>2σ(I) Goodness-of-fit on F Largest diff. peak and hole [e Å -3 ] w=1/[σ 2 (F o 2 )+(0.0116P) P] Weighting scheme 1.32 / where P=(F 2 o +2F 2 c )/3 30

31 Absorbance Absorbance 3. Stability in aqueous solution by UV Vis spectroscopy a 1% DMF / buffer a 1% DMSO / buffer Wavelength [nm] Wavelength (nm) Figure S15. UV Vis spectra of 2a in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h b 1% DMF / buffer b 1% DMSO / buffer Absorbance Absorbance Wavelength [nm] Wavelength [nm] Figure S16. UV Vis spectra of 2b in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h. 31

32 Absorbance Absorbance c 1% DMF / buffer c 1% DMSO / buffer Absorbance Absorbance Wavelength [nm] Wavelength [nm] Figure S17. UV Vis spectra of 2b in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h Absorbance d 1% DMF / buffer Absorbance d 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S18. UV Vis spectra of 2d in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h e 1% DMF / buffer e 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S19. UV Vis spectra of 2e in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h. 32

33 Absorbance Absorbance Absorbance a 1% DMF / buffer Absorbance a 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S20. UV Vis spectra of 3a in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h Absorbance b 1% DMF / buffer Absorbance b 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S21. UV Vis spectra of 3b in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h c 1% DMF / buffer c 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S22. UV Vis spectra of 3c in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h. 33

34 Absorbance Absorbance Absorbance d 1% DMF / buffer Absorbance d 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S23. UV Vis spectra of 3d in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h e 1% DMF / buffer e 1% DMSO / buffer Wavelength [nm] Wavelength [nm] Figure S24. UV Vis spectra of 2a in 1% DMF (left) or 1% DMSO (right) / phosphate buffer (ph 7.4), 1 h intervals over 24 h. 34

35 4. Stability toward aqueous phosphate buffer by NMR spectroscopy 24 h 1 h 0 h Ref. Figure S P NMR spectra of 2e in 10% d 7 -DMF / D 2 O containing 12 mm phosphate buffer at ph 7.4 after 0 h, 1 h, 24 h, and a reference spectrum without the addition of 2e. 24 h 1 h 0 h Ref. Figure S P NMR spectra of 3e in 10% d 7 -DMF / D 2 O containing 12 mm phosphate buffer at ph 7.4 after 0 h, 1 h, 24 h, and a reference spectrum without the addition of 3e. 35

36 24 h 1 h 0 h Ref. Figure S P NMR spectra of 2e in 10% d 6 -DMSO / D 2 O containing 12 mm phosphate buffer at ph 7.4 after 0 h, 1 h, 24 h, and a reference spectrum without the addition of 2e. 24 h 1 h 0 h Ref. Figure S P NMR spectra of 3e in 10% d 6 -DMSO / D 2 O containing 12 mm phosphate buffer at ph 7.4 after 0 h, 1 h, 24 h, and a reference spectrum without the addition of 3e.. 36

37 5. Stability and amino acid interaction via ESI-MS [M-Cl+DMSO] + [M-Cl] m/z 0 h 1 h 24 h t Figure S29. ESI-MS spectra after 0, 1 and 24 h of 2a in 1% DMSO / ammonium acetate solution (400 µm, ph 7.4), incubated at 37 C. [M-Cl+DMSO] + [M-Cl] m/z 0 h 1 h 24 h t Figure S30. ESI-MS spectra after 0, 1 and 24 h of 2a in 1% DMSO / ammonium acetate solution (400 µm, ph 7.4), incubated at 37 C with equimolar amount of His, Met and Cys. 37

38 Degree of M-Cl Hydrolysis [%] 6. Aquation-anation equilibria Degree of M-Cl Hydrolysis [%] without NaCl 5 mm NaCl 154 mm NaCl t [min] Figure S31. Degree of aquation of 2e as function of time at different NaCl concentration, expressed as the percentage of the aquated complex [M-Cl+OH 2 ] + in equilibrium with the chlorido complex [M] in 10% d 7 - DMF / D 2 O, by integration of ArH Ph-3 (aqua species, d at 8.43 ppm) and ArH Ph-4 (chlorido species, dd at 7.10 ppm) without NaCl 5 mm NaCl 154 mm NaCl t [min] Figure S32. Degree of aquation of 3e as function of time at different NaCl concentration, expressed as percentage of the aquated complex [M-Cl+OH 2 ] in equilibrium with the chlorido complex [M] in 10% d 7 - DMF / D 2 O, by integration of ArH Trz-5 (aqua species, s at ppm) and ArH Ph-3 (chlorido species, d at 8.12 ppm). 38

39 E: 1 M NaCl D: 154 mm NaCl C: 5 mm NaCl B: hydrolyzed, 0 mm NaCl A: partially hydrolyzed, 0 mm NaCl Figure S33. 1 H NMR spectra of complex 2e (0.5 mm) in 10% d 7 -DMF / D 2 O: (A) mixture of chlorido (squares, ) and aqua complex (triangles, ) 3 min after sample preparation; (B) complete hydrolysis after 30 min; addition of NaCl to a total concentration of 5 mm (C), 154 mm (D) and 1 M (E) respectively. 39

40 7. Topoisomerase IIα inhibition Neg control Marker/linear DNA Topo control VP16 - Poison ctrl DMF ctrl DMF w/o enzyme 1b 25 µm 1e 25 µm Ligands 2b 25 µm 2e 25 µm Ru complexes 3b 25 µm 3e 25 µm Os complexes Linear DNA Relaxed DNA No EtBr Supercoiled DNA Nicked, open circular Linear DNA Supercoiled DNA Relaxed DNA Containing EtBr Figure S34. Topoisomerase II drug screening assay. Negative control is phot DNA in water. Linear DNA is a marker. Topo control is phot DNA plus the enzyme. VP16 (etoposide) is the poison control. DMF control has a concentration of 0.625%. DMF without enzyme does not show any changes in DNA migration. 8. Cell cycle distribution Percentage of cells (%) e Concentration (µm) Percentage of cells (%) e Concentration (µm) Percentage of cells (%) e Concentration (µm) G2/M S G1 Figure S35. Cell cycle distributions (means of three independent experiments) in SW480 cells exposed to 1e (left), 2e (center) and 3e (right) for 48 h compared to untreated control (0 µm) at different concentrations. Application of higher concentrations than those indicated was impossible due to limited solubilities in most cases. 40

41 9. 1 H and 13 C NMR spectra Benzyl-4-phenyl-1,2,3-triazole (1a) Figure S36. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 1a in d 6 -DMSO. 41

42 (4 -Methoxybenzyl)-4-phenyl-1,2,3-triazole (1b) Figure S37. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 1b in d 6 -DMSO. 42

43 Propyl-4-phenyl-1,2,3-triazole (1c) Figure S38. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 1c in d 6 -DMSO. 43

44 Butyl-4-phenyl-1,2,3-triazole (1d) Figure S39. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 1d in d 6 -DMSO. 44

45 9.5. Methyl 2 -(4-phenyl-1,2,3-triazol-1-yl)acetate (1e) Figure S40. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 1e in d 6 -DMSO. 45

46 9.6. [Chlorido(1-benzyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazolato) (η 6 -pcymene)ruthenium(ii)] (2a) Figure S41. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2a in CDCl 3. 46

47 9.7. [Chlorido(1-(4 -methoxybenzyl)-4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazolato)(η 6 -p-cymene) ruthenium(ii)] (2b) Figure S42. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2b in CDCl 3. 47

48 9.8. [Chlorido(1-propyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazole)(η 6 -pcymene)ruthenium(ii)] (2c) Figure S43. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2c in CDCl 3. 48

49 9.9. [Chlorido(1-butyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazolato)(η 6 -pcymene)ruthenium(ii)] (2d) Figure S44. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2d in CDCl 3. 49

50 9.10. [Chlorido(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazol-1- yl)acetato)(η 6 -p-cymene)ruthenium(ii)] (2e) Figure S45. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2e in CDCl 3. 50

51 9.11. [Chlorido(1-benzyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazolato)(η 6 -pcymene)osmium(ii)] (3a) Figure S46. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3a in CDCl 3. 51

52 9.12. [Chlorido(1-(4 -methoxybenzyl)-4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazolato)(η 6 -p-cymene) osmium(ii)] (3b) Figure S47. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3b in CDCl 3. 52

53 9.13. [Chlorido(1-propyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazolato)(η 6 -pcymene)osmium(ii)] (3c) Figure S48. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3c in CDCl 3. 53

54 9.14. [Chlorido(1-butyl-4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazolato)(η 6 -pcymene)osmium(ii)] (3d) Figure S49. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3d in CDCl 3. 54

55 9.15. [Chlorido(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3-triazol-1- yl)acetato)(η 6 -p-cymene) osmium(ii)] (3e) Figure S50. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3e in CDCl 3. 55

56 9.16. [(κs-dimethylsulfoxide)(1-benzyl-4-(2 -κc)-phenyl-1,2,3-(3-κn)- triazolato)(η 6 -p-cymene)ruthenium(ii)] trifluoromethanesulfonate (2a ) Figure S51. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2a in CDCl 3. 56

57 9.17. [(κs-dimethylsulfoxide)(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazol-1-yl)acetato)(η 6 -p-cymene)ruthenium(ii)] trifluoromethanesulfonate (2e ) Figure S52. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 2e in CDCl 3. 57

58 9.18. [(κs-dimethylsulfoxide)(methyl-2 -(4-(2 -κc)-phenyl-(3-κn)-1,2,3- triazol-1-yl)acetato)(η 6 -p-cymene) osmium(ii)] trifluoromethanesulfonate (3e ). Figure S53. Atom labelling convention (top); 1 H (center) and 13 C (bottom) NMR of 3e in CDCl 3. 58