Metals and inorganic compounds have been used in medical therapies since the sixteenth century (Sadler, 1991; Abdel-Rahman, 2013; Abdel-Rahman, 2016) [1] [2] [3] . In 1969, the antitumor activity of metallic complexes containing cisplatin was discovered (Katsaros and Anagnostopoulou 2002) [4] enabling the development of new antitumor drugs such as complexes involving organic metals and/or inorganic platinum, ruthenium and rhodium (PAULA et al. 2005; Waxman and Anderson 2001) [5] [6] . Gallori and colleagues suggest that compounds containing ruthenium can bind to the DNA molecule, and that this interaction can alter the cell cycle, causing cell death through mechanisms such as apoptosis (Gallori et al. 2000) [7] . Evidence also suggests that several ruthenium complexes interact with specific proteins. One such application is the use of red ruthenium, which is traditionally used as a cytological stain for electron microscopy. In this case, binding of the compound occurs in the anionic sites of calcium-binding proteins (Clarke 2003; Suriano et al. 2005) [8] [9] . It was observed that several compound Ru²⁺ and Ru³⁺ amine ligands tend to interact selectively the carbonyl and imino sites of biomolecules that do not protonam at neutral pH. This leaves the pairs of electrons (of Nitrogen) available to coordinate bond with metal ions. Accordingly, ruthenium complexes often bind to proteins and imidazole nitrogen purine nucleotide (Clarke et al. 1978; Gupta et al. 2011) [10] [11] . Vilanova-Costa investigated the action of the compound cis-[RuCl₂(NH₃)₄], which showed antitumor activity in vitro on human tumor cell lines and in vivo on a tumor mouse strain (Sarcoma-180 (S-180) (Pereira et al. 2014; Vilanova-Costa et al. 2014; Vilanova-Costa et al. 2015) [12] [13] [14] . However, despite its pharmacological potential, the activity of ruthenium against leishmaniasis has not been extensively studied, even though the formation of granulomas that is characteristic of chronic leishmaniasis is similar to that observed in solid tumors (Arrais-Silva et al. 2006) [15] . Works that demonstrate the action of ruthenium against L. Mexicana promastigotes indicate the possible therapeutic action of the compounds of ruthenium (Navarro et al. 2006) [16] . Using in vitro models, Navarro (Navarro et al. 2006) [16] also demonstrated that some compounds containing the metal ruthenium decrease the L. Mexicana promastigotes forms by between 36% and 49% after treatment for 48 hours at a dose of 10μM, with loss of movement, fission of parasitic forms and vacuolation abundant in the host cell. The present study reports the synthesis of [Ru(Cl)₃(H₂O)₂(gly)] and its characterization by UV-VIS analysis, IR, TG/DTG and TG/DTA. A biological analysis of the effectiveness of the ruthenium compound in treating an experimental model of leishmaniasis was also carried out.

Materials: For synthesis of analytical grade complex compounds, RuCl₃∙3H₂O and glycine (NH₂CH₂COOH) were used. Distilled water, ethyl ether (C₄H₁₀O) and absolute alcohol (C₂H₆O) were used as solvents.

Synthesis: For the synthesis of compound [Ru(Cl)₃(H₂O)₂(gly)], 300.0 mg (1.440 mmol) of RuCl₃∙3H₂O was solubilized in 9.0 mL of distilled water, then 1000 mg (13.32 mmol) of glycine was added to the mixture under constant stirring, protected from light. After solubilization, the solution was allowed to reflux for 4 hours. The solution was held under ambient cold for 24 hours for precipitation. The precipitates were then filtered, washed with ethanol, ether and dried under reduced pressure. The solution was cooled again to obtain a higher yield of the precipitate. The solid was again washed with ethanol, ether and dried under reduced pressure.

Cell culture and parasites: Primary mouse macrophages were obtained from normal BALB/c mice by peritoneal lavage (Barbiéri et al. 1993; Giorgio et al. 1998) [17] [18] . Leishmania Amazonensis (MHOM/BR/73/M2269) amastigote forms were isolated from active skin lesions of BALB/c mice as described previously (Barbiéri et al. 1993) [17] . The parasites were suspended in RPMI 1640 medium and used immediately after isolation.

Macrophage infection and assessment of intracellular parasites: Peritoneal macrophages were infected with L. Amazonensisamastigotes (5:1 parasites/host cell) for 1 h, as previously described (Colhone et al. 2004) [19] . After the interaction period, the cultures were washed to remove extracellular parasites and fresh medium was added to the cell culture and incubated at 35˚C, 5% of CO₂ for 24 h, as previously described (Degrossoli and Giorgio 2007) [20] . Alternatively, different concentrations of RuCl₃∙3H₂O and glycine were added to culture of infected cells. Intracellular parasite destruction was assessed by morphological examination. Briefly, in order to evaluate the percentage of infected macrophages and the number of amastigotes per macrophage, cells cultured on coverslips were stained with Giemsa 0.6% (Giorgio et al. 1998) [18] . Intracellular amastigotes, which are exclusively localized in parasitophorous vacuoles (Chang 1980) [21] , were examined microscopically at a magnification of 1000×. About 600 cells were counted per triplicate coverslip (Linares et al. 2000) [22] .

Cytotoxicity assay: The cytotoxicity of ruthenium to macrophages was tested using the MTT viability assay, after incubation of peritoneal-derived macrophages with 50 to 400 µg/mL of ruthenium for 24 hours. The formation of formazan was measured by adding 0.5 mg/mL of 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT; Molecular Probes, Eugene, OR, USA) and incubating the cultures at 37˚C in the dark. After 4 h the medium was removed, 200 uL of DMSO (dimethyl sulphoxide) was added per well and the absorbance was measured using an ELISA reader at 540 nm (LabsystemsMultiskan). As a negative control, cells were also incubated with the highest concentration of DMSO used for ruthenium solubilization (0.01%).

Data analysis: The results were expressed as mean ± standard deviation (SD). To compare the average values of the parameters, analysis of variance (ANOVA) followed by multiple comparisons of Tukey test were used, with a statistical significance threshold of 5% (p < 0.05). The BioEstat version 5.0 software was used for all analyses (Ayres et al. 2007) [23] .

Instrumental Methods: For analysis of UV-VIS and IR for the compound of ruthenium equipment of the company Perkin Elmer®, model Lambda 25 UV-VIS analysis for the region of visible and UV-Spectrum-100 for the mid-infrared region were used. For the analysis of TG/DTG and TG-DTA, 6.6204 mg of the compound [Ru(Cl)₃(H₂O)₂gly] was placed in aα-alumina crucible and 90 uL of the sample was heated to 800˚C at a heating rate of 20˚C/min under an atmosphere of dry air a flow rate of 100 mL/min; the SDT 2960 TA Instruments® equipment was used for this analysis.

Ultraviolet visible analysis: Figure 1 shows the UV-Vis spectrum of the compound [Ru(Cl)₃(H₂O)₂(gly)] in water indicating the band 290 nm. The literature shows that ruthenium-glycine compounds exhibit absorption in the 290 nm region, little intense, as characterized as LMCT-type interactions (Load Transfer Ligand Metal) between the oxygen of the carboxyl group (glycine) and metallic ion (ruthenium) (Yeh and Taube 1980) [24] . the band found in the ultraviolet region at 230 nm, can be attributed to internal transitions of the ligand (IL), and were very similar in intensity to electronic transitions π - π* free ligand (Bento and Tfouni 1988) [25] .

Analysis in infrared medium region: Table 1 highlights the values of the vi-