Silver(I) metallodrugs of thiosemicarbazones and naproxen: biocompatibility, in vitro anti-proliferative activity and in silico interaction studies with EGFR, VEGFR2 and LOX receptors.

Four new heteroleptic silver(I) complexes with the general formula [Ag(L1-4)(nap)] (1-4), where L1-4 =  2-(1-(4-substitutedphenyl)ethylidene)hydrazinecarbothioamide and nap = naproxen, have been synthesized and characterized. The geometric parameters determined from density functional theory and UV-Vis studies indicate distorted tetrahedral geometry around silver(I) ion. Fourier transform infrared (FT IR) spectra evidenced asymmetric bidentate coordination mode of carboxyl oxygen atoms of naproxen with silver(I) ion. The complexes are stable for 72 h and biocompatibility was analysed towards normal human dermal fibroblast cells, which showed non-toxic nature up to 100 ng/ml. In vitro anti-proliferative activity of the complexes by MTT assay was tested against three human cancerous cell lines and one non-tumorigenic human breast epithelial cell line (MCF-10a) in which the complex 4 exhibited enhanced activity. The morphological changes observed by acridine orange/ethidium bromide and Hoechst 33258 staining method reveal apoptosis-inducing ability of the complexes. The molecular docking studies suggest hydrogen bonding, hydrophobic and π-pair interactions with the active site of epidermal growth factor receptor, vascular endothelial growth factor receptor 2 and lipoxygenase receptors.

Biocompatibility, in Vitro Antiproliferative, and in Silico EGFR/VEGFR2 Studies of Heteroleptic Metal(II) Complexes of Thiosemicarbazones and Naproxen.

Eight heteroleptic nickel(II) and copper(II) complexes of the type [M(L1-4)(nap)2] (1-8), where L1-4 = 2-(1-(4-substitutedphenyl)ethylidene)hydrazinecarbothioamide, nap = naproxen, and M = Ni(II) or Cu(II), have been synthesized and characterized. UV-vis and EPR spectral studies showed distorted octahedral geometry around metal(II) ions. The cyclic voltammogram of complexes 1-8 displayed an irreversible one-electron transfer process in the cathodic region (Epc = -0.66 to -1.43 V), and nickel(II) complexes 1-4 displayed an irreversible one-electron oxidation process in the anodic region (Epa = 0.75 to 1.10 V). The obtained magnetic moment values (1.82-1.93 µB) for copper(II) complexes 5-8 indicate distortion from octahedral geometry, which is further supported by EPR studies. The geometry of the complexes is retained in both solid and solution phases as evidenced from UV-vis and EPR studies. All the complexes showed stability for almost 72 h in biologically relevant solutions. The reducing ability of the copper(II) complexes in the presence of ascorbic acid was analyzed by UV-vis and cyclic voltammetry techniques, which indicates the reduction of the copper(II) to a copper(I) center, and possible interaction within the cells. An in vitro antiproliferative study revealed the nontoxic nature of complexes to normal human dermal fibroblast (NHDF) up to a concentration of 100 ng/mL. The antiproliferative activity of the complexes was tested against three cancerous (human breast adenocarcinoma (MCF-7), hepatoma (HepG2), and lung (A549)) cell lines using MTT reduction assay, which showed enhanced activity for complexes 4 and 8 containing the hydrophobic substituent. Apoptotic and cellular uptake studies showed that complex 8 is readily taken up by HepG2 cell lines and induces ROS-mediated mitochondrial and caspase-dependent apoptosis. In silico studies indicated hydrogen bonding, hydrophobic, and π-pair (π-π, π-σ, and π-cation) interactions between the complexes and EGFR/VEGFR2 kinase receptors.

Copper complexes as prospective anticancer agents: in vitro and in vivo evaluation, selective targeting of cancer cells by DNA damage and S phase arrest.

A series of six new bis(thiosemicarbazone)copper(i) complexes of the type [Cu(L1-6)2Cl] (1-6) have been synthesized and characterized. The molecular structure of the ligand L4 was determined by the single crystal XRD method. All the complexes adopted trigonal planar (Y-shaped) geometry. All the complexes strongly bind with CT-DNA via intercalative mode, which was further supported by molecular docking studies. Further, the complexes were effectively bind with BSA as observed by UV-Vis and fluorescence spectra. All the complexes effectively cleave pBR322 DNA through hydrolytic pathway as evidenced from T4 ligase experiments. All the complexes interact with the anticancer receptor focal adhesion kinase (FAK) via electrostatic, van der Waals, hydrogen bonding, σ-π and π-π interactions. In vitro cytotoxicity of the complexes were assessed by MTT assay against four cancer cell lines such as human breast adenocarcinoma (MCF-7), cervical (HeLa), epithelioma (Hep-2) and Ehrlich ascites carcinoma (EAC), and two normal cell lines namely normal human dermal fibroblasts (NHDF) and L6 myotubes with respect to the commercially used anticancer drug cisplatin. All the complexes induce apoptosis in EAC cells, which was confirmed by AO/EB, Hoechst 33258 and PI staining methods. The complexes block cell cycle progression of EAC cells in S phase (DNA synthesis). The cellular uptake studies confirmed the ability of the complexes to go into the cytoplasm and accumulation in the cell nuclei. In the in vivo anticancer studies, the complexes significantly reduce the tumour volume in female Swiss albino mice. Overall, our results ensure the role of thiosemicarbazone-based copper(i) complexes as prospective anticancer agents, induction of apoptosis and S phase arrest with the mitochondrial controlled pathway.

In vitro and in vivo anti-proliferative evaluation of bis(4'-(4-tolyl)-2,2':6',2″-terpyridine)copper(II) complex against Ehrlich ascites carcinoma tumors.

The bis(4'-(4-tolyl)-2,2':6',2″-terpyridine)copper(II) complex [Cu(ttpy)2]Cl2 was synthesized and authenticated by single crystal analysis, which shows distorted octahedral geometry around copper(II) ion. The crystal packing is stabilized by C-H···π inter and intramolecular interactions. The complex was found to be lipophilic as determined by shake-flask method. In vitro cytotoxicity of the complex was tested against Ehrlich ascites carcinoma (EAC) and L6 myotube cell lines. The complex exhibit potent cytotoxicity with respect to the commercially available anticancer drug cisplatin. Hoechst 33258, AO/EB and PI (flow cytometry) staining methods suggest that the complex can induce apoptosis in EAC cells. Cell cycle analyses also support the induced apoptosis. Cellular uptake studies revealed that the complex can go into the cytoplasm and accumulate in the cell nuclei. The complex induces EAC cell apoptosis through a ROS-mediated mitochondrial pathway by activating caspase 3 and caspase 7 and regulates the Bcl-2 family proteins. In vivo study of the complex was validated against the animal tumor growth (EAC) cell in Swiss albino mice. The bis(4'-(4-tolyl)-2,2':6',2″-terpyridine)copper(II) complex induces EAC cell apoptosis through a ROS-mediated mitochondrial pathway and significantly reduced the body weight and solid tumor volume in Swiss albino mice.

Heteroleptic silver(I) complexes with 2,2':6',2″-terpyridines and naproxen: DNA interaction, EGFR/VEGFR2 kinase, growth inhibition and cell cycle arrest studies.

A series of heteroleptic silver(I) complexes of the type [Ag(L1-3)(nap)] (1-3), where L1-3=4'-(4-substituted)-2,2':6',2″-terpyridines and nap=naproxen have been synthesized and characterized by elemental analysis and spectroscopic methods. The geometric parameters of the complexes were determined using UV-Vis and DFT calculations together with the IR spectral data. All the complexes adopted distorted tetrahedral geometry around silver(I) ion. The small HOMO-LUMO energy gap supports the bioefficacy of the complexes. DNA binding and melting experiments suggest the intercalative binding mode of the complexes to CT-DNA, which was further supported by molecular docking studies. The molecular docking studies of the heteroleptic silver(I) complexes with EGFR/VEGFR2 kinase receptors show hydrophobic, π-π, σ-π and hydrogen bonding interactions. All the complexes have been found to promote DNA cleavage through hydrolytic pathway. In vitro cytotoxicity activity of the complexes was tested against four human breast adenocarcinoma (MCF-7), cervical (HeLa), epithelioma (Hep-2) and hepatoma (HepG2) cancerous, and one normal human dermal fibroblasts (NHDF) cell lines by MTT reduction assay. The morphological study by Hoechst 33258 staining revealed that the complex 3 induces apoptosis much more effectively than the other complexes. Further, all the complexes increases the DNA synthesis in S phase with the corresponding reduction in G0-G1 and G2/M phase, which suggest the growth inhibition mechanism on HepG2 cells was DNA damage mediated S phase arrest.

Targeting of DNA molecules, BSA/c-Met tyrosine kinase receptors and anti-proliferative activity of bis(terpyridine)copper(ii) complexes.

A series of homoleptic bis(terpyridine)copper(ii) complexes of the type [Cu(L(1-5))2]Cl2 (), where L(1-5) = 4'-(4-substituted)-2,2':6',2''-terpyridines, have been synthesized and characterized. The molecular structure of complex was confirmed by the single crystal XRD technique, and the geometry of the complexes is best described as distorted octahedral. Structural parameters from the crystallographic and DFT studies are in good agreement with each other. The small HOMO-LUMO energy gap supports bioefficacy of the complexes. DNA binding studies show high intrinsic binding constant values 1.53 ± 0.15, 1.62 ± 0.08 and 3.09 ± 0.12 × 10(5) M(-1) for complexes , and , respectively, with intercalative mode of binding to CT-DNA. The binding results were further supported by molecular docking studies. The experimental results indicate that the interaction between the complexes and BSA protein involves a static quenching mechanism. The molecular docking studies with c-Met tyrosine kinase receptors show hydrophobic and π-π interactions. All the complexes bring about hydroxyl radical mediated DNA cleavage in the presence of H2O2. In vitro cytotoxicities of the complexes () were tested against three cancerous cell lines, namely human breast adenocarcinoma (MCF-7), epithelioma (Hep-2) and cervical (HeLa) cell lines, and one non-tumorigenic human dermal fibroblast (NHDF) cell line by MTT reduction assay. The morphological assessment data obtained using Hoechst 33258 staining revealed that complex induces apoptosis much more effectively than the other complexes.

Beneficial effects of methanolic extract of Indigofera linnaei Ali. on the inflammatory and nociceptive responses in rodent models

ABSTRACT Indigofera linnaei Ali. (Tamil Name: Cheppu Nerinjil) belongs to the family Fabaceae, used for the treatment of various ailments in the traditional system of medicine. In the present study, the beneficial effects of methanol extract of whole plant of I. linnaei (MEIL) were evaluated on inflammation and nociception responses in rodent models. In vitro nitric oxide (NO), lipoxygenase (LOX) and cyclooxygense (COX) inhibitory activities were also performed to understand the mode of action. MEIL at the dose of 200 & 400 mg/kg, p.o. significantly inhibited carrageenan induced rat paw volume and reduced the weight of granuloma in cotton pellet granuloma model. The results obtained were comparable with the standard drug aceclofenac. The anti-nociceptive effect of MEIL in mice was evaluated in hot plate and acetic acid induced writhing model. The plant extract significantly reduced the number of writhes and the analgesic effect was higher than that of the standard drug aspirin. However, the extract fails to increase the latency period in hot plate method suggesting that the extract produce nociception by peripheral activity. The extract produced inhibitory effect on NO, LOX and COX in concentration dependent manner. The extract exhibited pronounced and selective COX-2 inhibition. Altogether, these results suggested that the methanol extract of Indigofera linnaei could be considered as a potential anti-inflammatory and analgesic agent.

RESUMO Indigofera linnaei Ali pertence à família Leguminosae e é utilizada para o tratamento de várias doenças na medicina tradicional. No presente estudo, os efeitos benéficos do extrato metanólico da planta inteira de I. linnaei (MEIL) foram avaliados em respostas inflamatórias e nocicepção em modelos de roedores. Testes in vitro de atividade inibitória do óxido nítrico (NO), lipoxigenase (LOX) e ciclooxigenase (COX) também foram realizados para compreender o modo de ação. MEIL nas doses de 200 e 400 mg/kg, p.o. inibiu significativamente o volume da pata de rato induzido por carragenana e reduziu o peso do granuloma no modelo de pélete de algodão. Os resultados obtidos foram comparáveis ao do fármaco padrão, aceclofenaco. O efeito anti-nociceptivo de MEIL foi avaliado em camundongos no modelo de placa quente e de contorção induzida por ácido acético. O extrato da planta reduziu significativamente o número de contorções e o efeito analgésico foi maior do que o do fármaco padrão, ácido acetilsalicílico. Porém, o extrato não conseguiu aumentar o período de latência no método da placa quente, sugerindo que este produz nocicepção por atividade periférica. O extrato produziu efeito inibitório sobre o NO, LOX e COX dependente da concentração. O extrato exibiu inibição acentuada e seletiva da COX-2. No seu conjunto, estes resultados sugerem que o extrato metanólico de Indigofera linnaei poderia ser considerado como agente anti-inflamatório e analgésico potencial.

Antimicrobial efficacy and phytochemical analysis of Indigofera trita Linn.

An in vitro antimicrobial activity and phytochemical analysis of various extracts of Indigofera trita L. viz. petroleum ether, chloroform, acetone, ethanol and aqueous extracts were carried out. A total of 21 microorganisms (19 bacteria and 2 fungal strains) were used for antimicrobial activity by disc diffusion method and a standard procedure was used to identify the phytochemical constituents. Petroleum ether extract showed moderate inhibitory activity against Staphylococcus aureus (14.40 mm), S. epidermidis (14.20 mm), Salmonella paratyphi A (12.80 mm), Streptococcus mutans (12.20 mm), Escherichia coli, Proteus vulgaris, S. typhi and Burkholderia cepacia (12.00 mm). The chloroform extract also showed antimicrobial activity against S. epidermidis (14.20 mm), S. typhimurium (12.60 mm), S. paratyphi A, S. brunei and Yersinia enterocolitica (12.00 mm). The acetone extract of I. trita showed considerable inhibitory activity against S. epidermidis (18.20 mm), S. typhimurium (14.60 mm), S. infantis (13.80 mm), S. aureus (13.40 mm), Y. enterocolitica (13.00 mm) and Enterobacter aerogenes (12.00 mm) were documented. Ethanol extract showed significant antimicrobial activity against S. epidermidis (18.60 mm), S. paratyphi A (14.60 mm), Y. enterocolitica (13.40 mm), S. typhi (12.40 mm), S. aureus, E. aerogenes, S. typhimurium and S. infantis (12.00 mm). Aqueous extract of I. trita considerably inhibited S. epidermidis (13.80 mm), S. paratyphi A and Y. enterocolitica (12.20 mm), E. aerogenes and Haemophilus parahaemolyticus (12.00 mm). All the five extracts showed a minimal antifungal activity when compared to antibacterial activity. The result revealed that the antimicrobial properties of I. trita might be associated with the presence of phenolic compounds, flavonoids, tannins, glycosides, saponins, phytosterols and alkaloids.

In vitro and in vivo anticancer activity of Indigofera cassioides Rottl. Ex. DC.

OBJECTIVE: To evaluate the antitumor, cytotoxic and antioxidant activities of methanolic leaf extract of Indigofera cassioides (MEIC) against transplantable tumors and human cancer cell lines. METHODS: MEIC was investigated for its short term cytotoxicity on EAC and DLA cells by trypan blue dye exclusion method and in vitro cytotoxicity on HeLa, HEp-2, HEpG-2, MCF-7, HT-29, Vero and NIH 3T3 cells by MTT assay. In vivo antitumor activity was studied on EAC and DLA tumor bearing mice. Activity was assessed by monitoring the mean survival time, effect on hematological parameters, antioxidant enzyme levels and solid tumor volume. RESULTS: MEIC exhibit potent in vitro cytotoxicity against all the tested cancer cell lines, but it was found to be safe on normal cells. The extract significantly (P < 0.001) increase the mean survival time and also have a protective effect on the hemopoietic system at the tested dose levels (200 and 400 mg/kg). The extract prevented lipid peroxidation and restored the antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase in the liver of tumor control animals. It also significantly (P < 0.01) reduce the solid tumor volume. CONCLUSIONS: The results strongly support that MEIC shows potent antitumor and cytotoxic effects against EAC, DLA and human cancer cell lines. The extract prevents lipid peroxidation and promotes the enzymatic antioxidant defense system in tumor bearing animals which might be due to activities like scavenging of free radicals by the phytochemicals in MEIC.

Antitumor and cytotoxic activities of methanol extract of Indigofera linnaei Ali.

Methanol extract of Indigofera linnaei (MEIL) was investigated for antitumor, cytotoxic and antioxidant activities against transplantable tumors and human cancer cell lines. In vitro cytotoxicity was evaluated in HeLa, Hep-2, HepG-2, MCF-7, HT-29, Vero and NIH 3T3 cells by MTT assay and in vivo antitumor activity with Ehrlich ascites carcinoma (EAC) and Dalton's ascites lymphoma (DLA) tumor-bearing mice. Activity was measured by monitoring the mean survival time, effect on hematological parameters, antioxidant enzyme levels and solid tumor volume. The extract exhibited strong in vitro cytotoxicity against all the tested cancer cell lines, but it was found to be safe with normal cells. MEIL at the dose of 200 and 400 mg/kg, significantly increase the mean survival time (P<0.001), exerted a protective effect on the hemopoietic system, demonstrated in vivo antioxidant activity and significantly reduce solid tumor volume (P<0.01). These results show a significant antitumor and cytotoxic effect of MEIL against EAC, DLA and human cancer cell lines and support the ethnomedical use of Indigofera linnaei.

Effect of ethanol extract of flowers of Vitex trifolia Linn. on CCL4 induced hepatic injury in rats.

Hepatoprotective activity of ethanolic extract of flowers of Vitex trifolia (Verbenaceae) was studied against CCl4 induced hepatic injury in albino rats. The plant extract (EVT) at the dose of 200 mg/kg, p.o. showed a remarkable hepatoprotective activity. CCl4 induced a significant rise in serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), total bilirubin and gamma glutamate transpeptidase (GGTP). Treatment of rats with EVT significantly (P<0.001) altered serum biomarker enzyme levels to near normal against CCl4 treated rats. The activity of the extract was comparable to the standard drug, silymarin (100 mg/kg, p.o.). Histopathological observations also revealed that treatment with EVT protected the animals from CCl4 induced liver damage. The results indicate that the flowers of V. trifolia possess hepatoprotective activity on CCl4 induced hepatic injury in rats.