Supplementary Data
Biological Evaluation of Water Soluble Arene Ru(II) Enantiomers with Amino-Oxime
Ligands
Isabel de la Cueva-Aliqueǂ Sara Sierraǂ Laura Muntildeoz-Morenodagger Adriaacuten Peacuterez-Redondoǂ Ana M
Bajo dagger Isabel Marzo Lourdes Gudeǂ Tomaacutes Cuencaǂ Eva Royoǂ
ǂ Departamento de Quiacutemica Orgaacutenica y Quiacutemica Inorgaacutenica Instituto de Investigacioacuten Quiacutemica
Andreacutes M del Riacuteo (IQAR) Universidad de Alcalaacute 28805 Alcalaacute de Henares Madrid Spain
Departamento de Bioquiacutemica y Biologiacutea Molecular y Celular Universidad de Zaragoza 50009
Zaragoza Spain
dagger Departamento de Biologiacutea de Sistemas Facultad de Medicina y Ciencias de la Salud Universidad de
Alcalaacute 28805 Alcalaacute de Henares Madrid Spain
S1
Selected experimental data
1 Elemental analysis of compound 1a
2 Figures S1-S2 NMR spectra of proligands a and arsquo
3 Figures S3-S8 NMR spectra of 1a and 1arsquo
4 Figures S9 Time-dependent 1H NMR of 1arsquo in PBSD2O
5 Figures S10 UV-vis spectra of arsquomiddotHCl in water
6 Figure S11 UV-vis spectrum of 1arsquo in water
7 Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water
Selected crystallographic data
1 Table S1 Selected crystallographic data for 1arsquo
2 Figure S13 ORTEP drawing of 1arsquo cation
3 Table S2 Selected bond and angles for 1arsquo
Selected biological data
1 Figure S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) on A-549
cells
2 Figure S15 Effect of compounds 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability
3 Figure S16 FRET DNA melting curves with a∙HCl 1a and arsquo∙HCl 1arsquo at 10 M
4 Figure S17 Effect of treatment of PC-3 cells with 1a on the tumor growth
S2
Elemental analysis CHN for 1a
Anal Calcd for C27H38Cl2N2ORu C 5605 H 662 N 484 Found C 5598 H
667 N 479
S3
Selected NMR spectra
Figure S11H NMR of a in CDCl3
Figure S21H NMR of arsquo in CDCl3
S4
Figure S31H NMR of 1a in CD3OD
Figure S41H NMR of 1arsquo in CD3OD
S5
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Selected experimental data
1 Elemental analysis of compound 1a
2 Figures S1-S2 NMR spectra of proligands a and arsquo
3 Figures S3-S8 NMR spectra of 1a and 1arsquo
4 Figures S9 Time-dependent 1H NMR of 1arsquo in PBSD2O
5 Figures S10 UV-vis spectra of arsquomiddotHCl in water
6 Figure S11 UV-vis spectrum of 1arsquo in water
7 Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water
Selected crystallographic data
1 Table S1 Selected crystallographic data for 1arsquo
2 Figure S13 ORTEP drawing of 1arsquo cation
3 Table S2 Selected bond and angles for 1arsquo
Selected biological data
1 Figure S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) on A-549
cells
2 Figure S15 Effect of compounds 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability
3 Figure S16 FRET DNA melting curves with a∙HCl 1a and arsquo∙HCl 1arsquo at 10 M
4 Figure S17 Effect of treatment of PC-3 cells with 1a on the tumor growth
S2
Elemental analysis CHN for 1a
Anal Calcd for C27H38Cl2N2ORu C 5605 H 662 N 484 Found C 5598 H
667 N 479
S3
Selected NMR spectra
Figure S11H NMR of a in CDCl3
Figure S21H NMR of arsquo in CDCl3
S4
Figure S31H NMR of 1a in CD3OD
Figure S41H NMR of 1arsquo in CD3OD
S5
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Elemental analysis CHN for 1a
Anal Calcd for C27H38Cl2N2ORu C 5605 H 662 N 484 Found C 5598 H
667 N 479
S3
Selected NMR spectra
Figure S11H NMR of a in CDCl3
Figure S21H NMR of arsquo in CDCl3
S4
Figure S31H NMR of 1a in CD3OD
Figure S41H NMR of 1arsquo in CD3OD
S5
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Selected NMR spectra
Figure S11H NMR of a in CDCl3
Figure S21H NMR of arsquo in CDCl3
S4
Figure S31H NMR of 1a in CD3OD
Figure S41H NMR of 1arsquo in CD3OD
S5
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S31H NMR of 1a in CD3OD
Figure S41H NMR of 1arsquo in CD3OD
S5
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S51H NMR of 1arsquo in CDCl3
Figure S6 1H NMR of 1arsquo in D2O
S6
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S713C-APT NMR of 1arsquo in CD3OD
S7
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S815N-1H HMBC of 1arsquo in CD3OD
Figure S9 Time dependent 1H NMR of 1arsquo in a PBS solution (D2O) at pH = 740
S8
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
S9
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S10 UV-vis spectrum of arsquomiddotHCl in water (500M)
Figure S11 UV-vis spectrum of 1arsquo in water (500M)
S10
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S12 Circular dichroism spectra of amiddotHCl and arsquomiddotHCl in water (500 M)
-1
-08
-06
-04
-02
0
02
04
06
08
220 240 260 280 300 320Δε
λ (nm)
amiddotHCl amiddotHCl
S11
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Table S1 Crystallographic data for complex 1arsquo
Compound 1arsquomiddot2CHCl3
Empirical formula C29H40Cl8N2ORu
Formula weight 81730
Temperature (K) 200(2)
Wavelength (Aring) 071073
Crystal system Monoclinic
Space group P21
a (Aring) (ordm) 8865(2)
b (Aring) (ordm) 21916(2) 9049(1)
c (Aring) (ordm) 18183(2)
Volume (Aring3) 35327(8)
Z 4
Density calculated (g cmndash3) 1537
Absorption coefficient (mmndash1) 1075
F(000) 1664
Crystal size (mm3) 031 times 013 times 012
Theta range for data collection (ordm) 300 to 2524
Index ranges ndash10 to 10 ndash26 to 26ndash21 to 21
Reflections collected 66843
Independent reflections 12759 [R(int) = 0174]
Reflections [Igt2(I)] 8747
Completeness to theta 998
Goodness-of-fit on F2 1085
Final R indices [Igt2(I)] R1 = 0071 wR2 = 0114
R indices (all data) R1 = 0132 wR2 = 0134
Largest diff peak and hole (emiddotAringndash3) 0699 and ndash0588
S12
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S13 ORTEP drawing of the cation in the second independent molecule of the asymmetric unit
for compound 1arsquo with 50 probability ellipsoids Hydrogen bonded to carbon atoms have been
omitted for clarity
Table S2 Selected lengths (Aring) and angles (ordm) of the two independent molecules of the asymmetric
unit for 1arsquo
Ru(1)-Cl(1) 2404(4) Ru(1)-N(11) 2067(10)
Ru(1)-N(12) 2173(10) N(11)-O(11) 1381(13)
N(11)-C(21) 1284(15) N(12)-C(22) 1496(16)
C(21)-C(22) 1503(16) Ru(1)-Ct(1)a 1674
Ru(2)-Cl(3) 2396(4) Ru(2)-N(21) 2065(11)
Ru(2)-N(22) 2156(11) N(21)-O(21) 1388(13)
N(21)-C(61) 1263(16) N(22)-C(62) 1508(17)
C(61)-C(62) 1504(17) Ru(2)-Ct(2)b 1700
Cl(1)-Ru(1)-N(11) 829(3) Cl(1)-Ru(1)-N(12) 818(3)
N(11)-Ru(1)-N(12) 758(4) Ru(1)-N(11)-C(21) 1216(8)
N(11)-C(21)-C(22) 1157(11) C(21)-C(22)-N(12) 1087(10)
Ru(1)-N(12)-C(22) 1111(7) Ru(1)-N(12)-C(31) 1206(8)
C(22)-N(12)-C(31) 1127(10) Ru(1)-N(11)-O(11) 1240(8)
C(21)-N(11)-O(11) 1144(10) Cl(3)-Ru(2)-N(21) 820(3)
Cl(3)-Ru(2)-N(22) 820(3) N(21)-Ru(2)-N(22) 751(4)
Ru(2)-N(21)-C(61) 1225(9) N(21)-C(61)-C(62) 1156(12)
C(61)-C(62)-N(22) 1075(10) Ru(2)-N(22)-C(62) 1114(7)
Ru(2)-N(22)-C(71) 1204(9) C(62)-N(22)-C(71) 1113(10)
Ru(2)-N(21)-O(21) 1236(8) C(61)-N(21)-O(21) 1139(10)a Ct(1) centroid of the C(11)-C(16) ring b Ct(2) centroid of the C(51)-C(56) ring
S13
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Fig S14 Morphological changes induced by compounds 1a and 1arsquo (25 microM) A549 cells were seeded and 24 h later compounds were added at the
indicated concentrations Cell morphology was evaluated by optical microscopy after 24 h
S14
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Fig S15 Effect of enantiomers 1a and 1arsquo on JurKatpLVTHM and Jurkat shBak cell viability after 24 h of exposure The results are expressed as
a percentage of live cells compared to control
S15
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Figure S16 FRET DNA melting curves at 10 M of A) aHCl and 1a B) arsquoHCl and 1arsquo The concentration of F10T is 02 M
S16
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-
Fig S17 Effect of treatment of PC-3 cells with 1a on the tumor growth Androgen-independent
prostate cancer cells were incubated in the absence or presence of 1a (25 microM) for 24 h The
cell suspension was mixed with Matrigelreg and injected subcutaneously into the right flank of
nude mice (5 x 106 cellsmouse) Ten mice were used in each group A representative tumor
sample from each group after 43 days treatment is shown P lt 005 P lt 0001 vs control
group
S17
- Z
-