Xin
Li
a,
Kirsten
Heimann
bc,
Fangfei
Li†
a,
Jeffrey M.
Warner
cd,
F.
Richard Keene
*cef and
J.
Grant Collins
*a
aSchool of Physical, Environmental and Mathematical Sciences, University of New South Wales, Australian Defence Force Academy, Canberra, ACT 2600, Australia. E-mail: g.collins@adfa.edu.au
bCollege of Marine & Environmental Sciences, James Cook University, Townsville, QLD 4811, Australia
cCentre for Biodiscovery and Molecular Development of Therapeutics, James Cook University, Townsville, QLD 4811, Australia
dCollege of Public Health, Medical and Veterinary Sciences, James Cook University, Townsville, QLD 4811, Australia
eCollege of Science, Technology & Engineering, James Cook University, Townsville, QLD 4811, Australia
fSchool of Physical Sciences, University of Adelaide, Adelaide, SA 5005, Australia. E-mail: Richard.keene@adelaide.edu.au
First published on 3rd February 2016
A series of non-symmetric dinuclear polypyridylruthenium(II) complexes (Rubbn-Cl) that contain one inert metal centre and one coordinatively-labile metal centre, linked by the bis[4(4′-methyl-2,2′-bipyridyl)]-1,n-alkane ligand (“bbn” for n = 7, 12 and 16), have been synthesised and their potential as antimicrobial agents examined. The minimum inhibitory concentrations (MIC) of the ruthenium(II) complexes were determined against four strains of bacteria − Gram-positive Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), and Gram-negative Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa). The Rubbn-Cl complexes displayed good antimicrobial activity, with Rubb12-Cl being the most active complex against both Gram-positive and Gram-negative strains. Interestingly, Rubb7-Cl was found to be eight- and sixteen-fold more active towards E. coli than against S. aureus and MRSA, respectively. The cytotoxicities of the Rubbn-Cl complexes against three eukaryotic cell lines – two kidney cell lines (BHK and HEK-293) and one liver cell line (HepG2) – were examined. The Rubbn-Cl complexes were found to be considerably less toxic towards eukaryotic cells than S. aureus, MRSA and E. coli, with Rubb12-Cl being thirty- to eighty-times more toxic to the bacteria than to BHK, HEK-293 or HepG2 cells. Unexpectedly, Rubb7-Cl was far more toxic to HepG2 cells (24 h-IC50 = 3.7 μM) and far less toxic to BHK cells (24 h-IC50 = 238 μM) than the Rubb12-Cl and Rubb16-Cl complexes. In order to understand the unexpected large differences in the cytotoxicities of the Rubbn-Cl complexes towards eukaryotic cells, a confocal microscopic study of their intracellular localisation was undertaken. The results suggest that the observed cytotoxicity might be related to the extent of DNA binding.
Traditionally, the design and development of new antimicrobial drugs has centred upon organic chemistry. However, due to the success of cisplatin as an anticancer drug and the established ability of transition metal complexes to bind DNA and RNA,2–6 there has been increasing interest in using metal complexes as antimicrobial agents.7–14 Among the transition metal complexes, ruthenium-based complexes have drawn increasing attention.9–14 Dwyer and co-workers were the first to report the biological activity of mononuclear tris(bidentate) inert polypyridyl metal complexes, in particular complexes with 1,10-phenanthroline ligands.9,10 [Ru(phen)3]2+ was found to be inactive; however, the introduction of methyl substituents on the phen ligands dramatically increased the activity against all bacteria.9,10 More recently, it has been demonstrated that polypyridylruthenium(II) complexes which bind DNA by intercalation have significant bactericidal activity, particularly against Gram-positive strains.11,12 While DNA binding is generally thought to be responsible for the antimicrobial activity of polypyridylruthenium(II) complexes, Lam et al. suggested that the antimicrobial activity of a bis(bipyridine)ruthenium(II) complex containing a N-phenyl-substituted diazafluorene ligand might be due to DNA damage caused by the formation of reactive oxygen species.15 In addition, a range of labile ruthenium(II) and ruthenium(III) complexes have shown antimicrobial activity.16,17
We have recently demonstrated that dinuclear polypyridylruthenium(II) complexes containing a flexible bis[4(4′-methyl-(-2,2′-bipyridyl)]-1,n-alkane (bbn) bridging ligand (see Fig. 1) have good antimicrobial activity.18–21 These ruthenium complexes were highly active against a range of pathogenic bacteria, particularly Gram-positive strains,18 and maintained the activity against drug-resistant bacteria, including strains that are of considerable current concern, e.g. methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE). Furthermore, preliminary toxicity experiments indicated the dinuclear Rubbn complexes were significantly less toxic to eukaryotic cells.22,23 Based upon the good antimicrobial activity and cell selectivity of the Rubbn complexes, corresponding tri- and tetra-nuclear inert ruthenium complexes were subsequently synthesised.24 These complexes generally showed better activities than the dinuclear analogues and were more active against Gram-positive species.24
In another approach, we have also examined the effects of incorporating labile chlorido groups into dinuclear ruthenium(II) complexes linked by the bbn ligand, [{Ru(tpy)Cl}2{μ-bbn}]2+ (Cl-Rubbn-Cl; where tpy = 2,2′:6′,2′′-terpyridine),25 see Fig. 2. The symmetrical Cl-Rubbn-Cl complexes showed good activity against both Gram-positive bacteria and Gram-negative species. However, incorporation of the chlorido groups did significantly affect the relative activities of the ruthenium complexes, compared to the corresponding inert Rubbn complexes. Whereas the order of activities for the inert complexes was Rubb16 ≥ Rubb12 > Rubb7, it was found that Cl-Rubb12-Cl was the most active of the Cl-Rubbn-Cl complexes and Cl-Rubb16-Cl was slightly less active than the Cl-Rubb7-Cl complex. Taken together, the combined results highlight the balance between cationic charge and lipophilicity; however, it is not yet clear where the optimal charge/lipophilicity balance lies. In order to help clarify this issue, we aimed to synthesise and examine the antimicrobial activities of dinuclear ruthenium complexes that contained one inert metal centre and one metal centre that incorporated a chlorido ligand (Rubbn-Cl complexes, see Fig. 2).
In this study, the synthesis and the antimicrobial properties of the non-symmetrical Rubbn-Cl complexes (for n = 7, 12 and 16) against Gram-positive S. aureus and MRSA, and Gram-negative Escherichia coli and Pseudomonas aeruginosa were examined. As the clinical potential of any new drug is determined by both the antimicrobial activity and the associated toxicity towards eukaryotic cells, the cytotoxicities of the Rubbn-Cl complexes against three eukaryotic cell lines were also examined. In order to understand the unexpected large differences in the cytotoxicities of the Rubbn-Cl complexes towards eukaryotic cells, a confocal microscopic study of their intracellular localisation was also undertaken.
The mononuclear ruthenium complex [Ru(tpy)(Me2bpy)Cl]Cl was synthesised as previously described.26 The synthesis of the bridging ligands bbn (n = 7, 12 and 16; A) were performed in a similar manner to that reported in the literature.27 The precursors [Ru(tpy)Cl3] (B) [Ru(phen)2Cl2]Cl (C), [Ru(phen)2Cl2] (D), [Ru(phen)2(py)2]Cl2 (E; py = pyridine) and rac-[Ru(phen)2(bbn)](PF6)2 (F) were prepared according to previously reported methods.27–29
[Ru(phen)2(μ-bb12)Ru(tpy)Cl]Cl3 and [Ru(phen)2(μ-bb16)Ru(tpy)Cl]Cl3 were synthesised using an analogous procedure as that for [Ru(phen)2(μ-bb7)Ru(tpy)Cl]Cl3 but with the appropriate bbn bridging ligand.
Δ-[Ru(phen)2(μ-bbn)Ru(tpy)Cl]Cl3 complexes were synthesised following the same procedure as that for the racemic mixtures, using Δ-[Ru(phen)2(bbn)](PF6)2 (n = 7, 12 and 16) as precursors. CD spectra: Δ-Rubb7-Cl {λ/nm (Δε/cm−1 M−1) H2O}: 290 (−374.3), 281 (−294.2), 272 (−405.1), 261 (469.3), 233 (27.3), 221 (77.5). Δ-Rubb12-Cl {λ/nm (Δε/cm−1 M−1) H2O}: 290 (−356.7), 281 (−244.3), 271 (−352.3), 261 (412.8), 228 (10.5), 213 (107.4). Δ-Rubb16-Cl {λ/nm (Δε/cm−1 M−1) H2O}: 288 (−364.4), 279 (−285.9), 271 (−358.7), 260 (393.6), 227 (39.2), 221 (80.5).
Molecular modelling was performed using HyperChem.31 Energy minimisation by Polak–Ribiere conjugate-gradient refinement was carried out with the metal complex treated as a rigid group. The ruthenium complex was manually docked to the GMP to reflect observed intermolecular NOEs.
The cellular localisation of the ruthenium complexes was determined using a Zeiss laser scanning confocal microscope (LSM 700, Carl Zeiss, Göttingen, Germany). Samples were viewed under a 63× oil immersion lens. The ruthenium complexes (λex = 450 nm, λem = 610 nm), Mitotracker Green FM (λex = 490 nm, λem = 516 nm) and SYTO 9 (λex = 486 nm, λem = 501 nm) were excited using a blue argon laser (λex = 488 nm), and emissions were collected over the range 570–650 nm for the metal complexes, 470–550 nm for Mitotracker and 495–510 nm for SYTO 9. For DAPI excitation, a diode laser (λex = 405 nm) was used and the emission detected at 430–500 nm. Image data acquisition and processing was performed using Zen software 2009 (Carl Zeiss).
Given the observed preferential binding of [Ru(tpy)(Me2bpy)(D2O)]2+ with GMP, further studies were only carried out with GMP. Fig. 5 shows the 1H NMR spectrum of [Ru(tpy)(Me2bpy)(D2O)]2+ with added GMP as a function of time, and after the addition of further GMP after 64 hours. The H8 of GMP was tentatively assigned by heating the sample at 50 °C for 16 hours. The H8 of GMP will slowly exchange with a deuterium in the D2O solvent at 50 °C; e.g. see the reduction in intensity of the H8 of the free GMP in Fig. 5D and E. Consequently, the singlet resonance at 6.56 ppm could be assigned to the H8 of bound GMP.
The assignment of the proton resonances from the [Ru(tpy)(Me2bpy)GMP] adduct was determined from DQFCOSY and NOESY spectra (data not shown). In the NOESY spectrum of the aromatic to sugar H1′ region, strong NOE cross peaks were observed between the tpy H6 and H6′′, H5 and H5′′, H4 and H4′′, H3 and H3′′ and H3′ and H5′, indicating the tpy protons were in slow exchange (on the NMR time scale) between two forms. Due to rotation around the Ru–N7 bond, it is possible that [Ru(tpy)(Me2bpy)GMP] complex may exist in two conformers that are in slow exchange. An NOE cross peak between the H8 and sugar H1′ of the bound GMP at 6.56/5.50 ppm provided further support for the assignment of the H8 of the bound GMP. Based on the NMR analysis, the binding site of ruthenium complex is most likely at the N7 of the GMP, given the large shift observed for the GH8 resonance. A molecular model of the ruthenium complex-GMP adduct is shown in Fig. 6. In the model, the GMP H8 is positioned directly below the tpy aromatic rings, which may explain the unusual upfield shift observed for the resonance upon N7-metallation.
The time-course 1H NMR spectra of the reaction between the dinuclear complex Rubb7-Cl and GMP also showed the emergence of resonances for the ruthenium complex-GMP adduct (data not shown). A singlet at 6.65 ppm could be assigned to the H8 from bound GMP, and new broad peaks in the 8.7–8.9 ppm region were possibly from the bound metal complex. These observations were similar to those from the reaction between [Ru(tpy)(Me2bpy)Cl]+ and GMP, described above, suggesting that Rubb7-Cl interacts with GMP by forming a covalent bond.
Complexes | S. aureus | MRSA | E. coli | P. aeruginosa | ||||
---|---|---|---|---|---|---|---|---|
MIC | MBC | MIC | MBC | MIC | MBC | MIC | MBC | |
Δ-Rubb7-Cl | 5.6 | 11.2 | 11.2 | 22.4 | 0.7 | 0.7 | 11.2 | 22.4 |
Δ-Rubb12-Cl | 0.7 | 0.7 | 0.7 | 1.4 | 0.7 | 0.7 | 11.2 | 11.2 |
Δ-Rubb16-Cl | 0.7 | 0.7 | 1.4 | 1.4 | 2.7 | 2.7 | 43.2 | 43.2 |
ΔΔ-Rubb7 | 10.7 | 21.3 | 10.7 | 21.3 | 10.7 | 10.7 | 85.3 | >85 |
ΔΔ-Rubb12 | 0.7 | 1.3 | 0.7 | 1.3 | 1.3 | 1.3 | 20.4 | 20.4 |
ΔΔ-Rubb16 | 0.6 | 0.6 | 0.6 | 1.2 | 1.2 | 1.2 | 9.8 | 9.8 |
Gentamicin | 0.4 | 0.8 | 28 | >200 | 0.8 | 0.8 | 1.6 | 3.2 |
Over the four bacteria examined in this study, Rubb7-Cl and Rubb12-Cl showed slightly better antimicrobial activities compared to their inert analogues, Rubb7 and Rubb12. Conversely, Rubb16-Cl displayed slightly lower activity than Rubb16. Interestingly, Rubb7-Cl and Rubb12-Cl show better activities against the two Gram-negative species E. coli and P. aeruginosa than Rubb7 and Rubb12, respectively. Particularly noteworthy is the activity of Rubb7-Cl against E. coli compared to the Gram-positive species S. aureus and MRSA: Rubb7-Cl is eight- and sixteen-fold more active towards E. coli than against S. aureus and MRSA, respectively. This preferential activity towards a Gram-negative species is very unusual for metal-based antimicrobial agents.
Complexes | IC50 | SI | ||||
---|---|---|---|---|---|---|
BHK | HEK-293 | BHK vs. S. aureus | BHK vs. E. coli | HEK-293 vs. S. aureus | HEK-293 vs. E. coli | |
Rac-Rubb7-Cl | 238.1 ± 14.8 | 69.1 ± 5.6 | 42.5 | 340.1 | 12.3 | 98.7 |
Rac-Rubb12-Cl | 47.4 ± 0.9 | 58.7 ± 2.1 | 67.7 | 67.7 | 83.9 | 83.9 |
Rac-Rubb16-Cl | 22.6 ± 2.3 | 41.1 ± 1.9 | 32.3 | 8.4 | 58.7 | 15.2 |
ΔΔ-Rubb12 | 70.5 ± 26.4 | 50.9 ± 19.9 | 100.7 | 54.2 | 72.7 | 39.2 |
ΔΔ-Rubb16 | 29.8 ± 1.1 | 21.0 ± 10.8 | 49.7 | 24.8 | 35 | 17.5 |
Generally, the complexes with a longer linking chain were more toxic to the cells. Among the rac-Rubbn-Cl complexes, Rubb16-Cl was the most toxic complex towards BHK and HEK-293 cells. In contrast, Rubb7-Cl was nontoxic (>200 μM) towards BHK but showed moderate cytotoxicity towards HEK-293, while Rubb12-Cl showed similar cytotoxicity towards both cell lines. These observations suggest that the linking chain length plays an important role in their cytotoxicities. The inert complexes ΔΔ-Rubb12 and ΔΔ-Rubb16 also showed the same trend with ΔΔ-Rubb16 being more toxic than ΔΔ-Rubb12.
The good antimicrobial activity of the ruthenium complexes suggests they may have potential as antimicrobial agents. However, to be clinically useful as antimicrobial agents, it is desirable that the compounds exhibit significantly greater toxicity towards bacterial cells than mammalian cells. Table 2 also summarises the selectivity indices (SI, 24 h-IC50/MIC) between the BHK and HEK-293 cell lines and the Gram-positive bacterium S. aureus and the Gram-negative species E. coli. In general, the SI values demonstrated that all the ruthenium complexes were more toxic against bacterial cells than eukaryotic cells, with the SI values ranging from 8–340. Noticeably, Rubb7-Cl displayed the highest SI values for both BHK and HEK-293 cells against E. coli. Of particular note, the SI value of 340 between BHK and E. coli was at least five-times higher than with the other complexes. By contrast, Rubb12-Cl showed similar selectivity towards S. aureus and E. coli, compared to both kidney cell lines. The inert complex Rubb12 displayed a better selectivity for S. aureus than E. coli against both kidney cell lines. Taken together, Rubb12-Cl exhibited the best overall selectivity (but only marginally better than Rubb12), while Rubb7-Cl showed the best selectivity towards the Gram-negative E. coli.
The liver cell line HepG2 was chosen to determine the cytotoxicities of the ruthenium complexes in liver cells. The cytotoxicities of the ruthenium complexes against HepG2 cells were determined after a 24-hour incubation, and the results are summarised in Table 3. The HepG2 cells were more susceptible than the kidney cells to Rubb7-Cl and Rubb12-Cl, but were more resistant to Rubb16-Cl. Interestingly, and unlike what was observed with the BHK and HEK-293 cells, the cytotoxicities of the Rubbn-Cl complexes decreased with increasing chain length in the bbn ligand. Surprisingly, Rubb7-Cl was the most cytotoxic of all the ruthenium complexes assayed against the HepG2 cells, with the 24 h-IC50 being 3.7 μM. The comparative selectivities between the two kidney cell lines and the HepG2 cell line were calculated for each of the ruthenium complexes and the results are summarised in Table 3. Rubb7-Cl clearly exhibited significantly higher toxicity towards the HepG2 cell line − compared to the two kidney cell lines − than did the other ruthenium complexes. Although the relative difference was much smaller, Rubb12-Cl was the only other complex to show some preferential toxicity to the HepG2 cells.
Complexes | IC50 | SX | |
---|---|---|---|
HepG2 | BHK vs. HepG2 | HEK-293 vs. HepG2 | |
Rac-Rubb7-Cl | 3.7 ± 0.7 | 64.4 | 18.7 |
Rac-Rubb12-Cl | 24.3 ± 3.9 | 2.0 | 2.4 |
Rac-Rubb16-Cl | 52.0 ± 2.9 | 0.4 | 0.8 |
ΔΔ-Rubb12 | 61.7 ± 5.5 | 1.1 | 0.8 |
ΔΔ-Rubb16 | 41.5 ± 2.9 | 0.7 | 0.5 |
As shown in Fig. 7, the rac-Rubbn-Cl complexes showed similar nucleolus localisation to that previously reported for the inert Rubbn complexes.22 The nucleolus of HepG2 cells were stained by Rubbn-Cl and SYTO 9. In addition to SYTO 9-stained rRNA, the most toxic complex Rubb7-Cl also showed localisation with DAPI-stained DNA. By contrast, Rubb12-Cl and Rubb16-Cl overlaid more with SYTO 9-stained rRNA in the nucleolus, and less with DAPI-stained DNA. In particular, Rubb12-Cl was exclusively localised in the nucleolus. These results suggest that Rubb12-Cl and Rubb16-Cl have greater selectivity for rRNA, compared to DNA, than does Rubb7-Cl.
BHK cells incubated with Rubb7-Cl at a concentration of 100 μM (approx. 50% of the IC50) for 20 hours are shown in Fig. 8. The Rubb7-Cl phosphorescence is observed throughout the cytoplasm and in the nucleus; however, within the nucleus the phosphorescence was mainly in the nucleolus, with little overlay with the DAPI-stained DNA. When BHK cells were incubated with Rubb7-Cl at a concentration of 50 μM for 20 hours, almost no phosphorescence was observed in the nucleus, with only relatively weak accumulation of the ruthenium complex in the cytoplasm (see Fig. 9).
![]() | ||
Fig. 9 DAPI (blue) and rac-Rubb7-Cl (red; 50 μM; 20 hour incubation) localisation in BHK cells. Scale bar = 10 μm. |
The preferential activity of Rubb7-Cl towards Gram-negative E. coli over the Gram-positive species S. aureus and MRSA is very unusual for a metal-based antimicrobial agent. A few metal complexes have shown higher activity against Gram-negative than Gram-positive bacteria. For example, two helicate-like dinuclear iron complexes showed two-fold higher activity against E. coli than S. aureus,35 and a [Ru2L3]4+ triply-stranded helicate complex also showed higher activity towards E. coli than S. aureus, but the activities were modest.36 However, most metal complexes have shown greater antimicrobial activity against Gram-positive species than Gram-negative species.9–11,18–25,37,38
The Rubbn-Cl complexes also showed a different pattern of activities to the Rubbn and Cl-Rubbn-Cl complexes. Against the Gram-positive bacteria, Rubb12-Cl and Rubb16-Cl were of equal activity and considerably more active than Rubb7-Cl. However, for the Gram-negative species, Rubb7-Cl and Rubb12-Cl were of equal activity and were considerably more active than Rubb16-Cl. For the Rubbn and Cl-Rubbn-Cl complexes, the same pattern of activities was observed for all bacteria. For the Rubbn complexes, the order of the activities across the four bacteria was Rubb16 ≥ Rubb12 > Rubb7; while for the Cl-Rubbn-Cl series, Cl-Rubb12-Cl > Cl-Rubb7-Cl ≥ Cl-Rubb16-Cl across the bacteria. Although the differences are relatively small, Rubb12-Cl showed the best activity profile against the four bacterial species used in this study compared to any of the bbn-linked ruthenium complexes that have been previously reported.18–25
Cytotoxicity assays against the kidney cell lines BHK and HEK-293 and the liver cell line HepG2 were carried out to estimate the toxicity of the Rubbn-Cl complexes towards mammalian organ cells. In BHK and HEK-293 cells, the 24 h-IC50 values for the Rubbn-Cl series decreased with the increasing number of methylene groups in the bbn ligand. A similar trend was observed for the Rubbn complexes. These results suggest that cellular uptake is the key parameter, at least to a first approximation, with the uptake being correlated to the lipophilicity of the ruthenium complex. However, in HepG2 cells the least lipophilic complex Rubb7-Cl exhibited the highest cytotoxicity while the most lipophilic complex Rubb16-Cl was the least toxic. Hence, it is probable that while lipophilicity is important, at least in terms of cellular uptake, there are other factors to be considered. Compared with healthy cells, the cancer cell outer membrane leaflet contains three- to nine-times more negatively-charged lipids and greater levels of negatively-charged O-glycosylated mucins.39–41 In contrast, in non-cancerous cells the membrane is largely occupied by zwitterionic phospholipids, with negligible or weak negative charge.42,43 In addition, the increased number of microvilli on cancer cells, which lead to an increase in cell surface area, may also enhance their susceptibility.44,45 Therefore, the cellular uptake of the polycationic ruthenium complexes may be less affected by lipophilicity in cancer cell lines compared with healthy cells.
Intracellular localisation could also be an important aspect of cytotoxicity. Confocal microscopy was used to determine the cellular localisation of the Rubbn-Cl complexes in HepG2 cells. The Rubbn-Cl complexes preferentially accumulated in the nucleolus, while significant DNA binding was also observed at higher concentrations. The preference for rRNA is consistent with our previous studies on the localisation of Rubb12 in BHK cells22 and Rubb16 in the ribosomes of E. coli.21 However, the Rubbn-Cl complexes showed differences in nuclear localisation in HepG2 cells based upon the length of the alkyl chain in the bbn linking ligand. The least lipophilic complex Rubb7-Cl appeared to co-localise to a higher degree with DNA, while the other two complexes, particularly Rubb12-Cl, showed greater accumulation in the nucleolus. The effect of lipophilicity on the cellular localisation of ruthenium complexes has been previously reported. Lincoln, Nordén and co-workers found the length of an alkyl chain in a dppz-based complex (dppz = dipyrido[3,2-a:2′,3′-c]phenazine) had a significant effect on the localisation pattern – the least lipophilic complex was found to stain nuclear DNA, the most lipophilic complex preferably stained cellular membranes, whereas the derivative of intermediate lipophilicity selectively stained the RNA-rich nucleoli.46 Furthermore, Thomas and co-workers demonstrated that the dinuclear complex [{Ru(phen)2}2{μ-tpphz}]4+ (where tpphz = tetrapyridophenazine) localised in the nucleus, while the more lipophilic 4,7-diphenyl-1,10-phenanthroline analogue [{Ru(DIP)2}2{μ-tpphz}]4+ localised in the endoplasmic reticulum.47,48 While other factors are yet to be examined, the greater level of DNA localisation in HepG2 by Rubb7-Cl, compared to Rubb12-Cl and Rubb12, could be related to its higher cytotoxicity against this cell line. Consistent with this proposal was the observed low level of DNA binding, compared to that in the nucleolus and cytoplasm, by Rubb7-Cl in BHK cells at concentrations considerably higher than the 24 h-IC50 determined against HepG2 cells.
In conclusion, a new class of dinuclear ruthenium complexes, Rubbn-Cl, has been synthesised and characterised. These ruthenium complexes exhibited good antimicrobial activities; and interestingly, showed relatively better activity towards Gram-negative bacteria, (compared to Gram-positive species) than previously reported ruthenium complexes linked by the bbn ligand. In addition, the Rubbn-Cl complexes were considerably less toxic to eukaryotic cells, compared to bacteria, with Rubb7-Cl showing striking differences in cytotoxicity between the eukaryotic cell lines. It is possible that Rubb7-Cl could become a new lead compound for metal-based anticancer or antimicrobial agents.
Footnote |
† Current address: School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China. |
This journal is © The Royal Society of Chemistry 2016 |