Arumugam Sudha

Exploration of potential EGFR inhibitors: a combination of pharmacophore-based virtual screening, atom-based 3D-QSAR and molecular docking analysis

Abstract Epidermal growth factor receptor (EGFR) protein tyrosine kinases are over expressed in several human cancers and considered as a promising target for developing novel anticancer drugs. In this study, the ligand-based pharmacophore mapping and atom-based 3D-QSAR approach was carried out on a series of 40 novel pyrrolo[3, 2-d]pyrimidine derivatives acting as EGFR inhibitors. The best pharmacophore hypothesis AAADRR.295 was selected and an atom-based 3D-QSAR model was generated by applying partial least-squares algorithm. The developed model was validated and used as a 3D query in sequential virtual screening study to filter five chemical databases. The obtained compounds were further filtered according to Lipinski rule of five and fitness score. Subsequently, a multistep molecular docking study was employed on the retrieved hits and finally, 12 compounds were prioritized as potential leads against EGFR, which exhibited high docking scores, correlated binding mode to experimentally proven compounds and constructive drug-like properties. The results of this study provide detailed structural insights and emphasize the important binding features of these compounds, which may assists in the design and development of novel EGFR inhibitors.

In vitro, fluorescence-quenching and computational studies on the interaction between lipoxygenase and 5-hydroxy-3',4',7-trimethoxyflavone from Lippia nodiflora L.

Abstract Lippia nodiflora L. is extensively used in traditional medicine for several medicinal purposes, including their use in inflammatory disorders. In this study, the folk use of L. nodiflora was validated using the isolated natural compound, 5-hydroxy-3′,4′,7-trimethoxyflavone (HTMF) by in vitro, fluorescence spectroscopic and molecular modeling studies with lipoxygenase (LOX), because LOX plays an essential role in inflammatory responses. In this perspective, the methanol extract and HTMF are shown to demonstrate prominent inhibitory activity against soybean lipoxygenase, with an IC50 value of 21.12 and 23.97 µg/ml, respectively. The data obtained from the spectroscopic method revealed that the quenching of intrinsic fluorescence of LOX is produced as a result of the complex formation of LOX–HTMF. The binding mode analysis of HTMF within the LOX enzyme suggested that hydrogen bond formation, hydrophobic interaction and π–π stacking could account for the binding of HTMF. Molecular dynamics results indicated the interaction of HTMF with LOX and the stability of ligand–enzyme complex was maintained throughout the simulation. The computational results are reliable with experimental facts and provided a good representation for understanding the binding mode of HTMF inside the active site of lipoxygenase enzyme.

Protective effect of 5-hydroxy-3′,4′,7-trimethoxyflavone against inflammation induced by lipopolysaccharide in RAW 264.7 macrophage: in vitro study and in silico validation

The herb Lippia nodiflora L. (Verbenaceae) has been documented to exhibit anti-inflammatory, antipyretic, antitussive, antidiabetic, anticancer, and antimelanogenesis properties. In the present study, we aimed at evaluating the anti-inflammatory activity of 5-hydroxy-3′,4′,7-trimethoxyflavone, a flavonoid from L. nodiflora, using lipopolysaccharide induced inflammation in RAW 264.7 macrophages. 5-hydroxy-3′,4′,7-trimethoxyflavone significantly inhibited nitric oxide production and demonstrated slight reduction in prostaglandin-E2 level at tested concentrations. The production of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-1β, were obviously reduced by 5-hydroxy-3′,4′,7-trimethoxyflavone in a concentration-dependent manner. Moreover, 5-hydroxy-3′,4′,7-trimethoxyflavone significantly induced reduction in the mRNA expressions of inducible nitric oxide synthase and cyclooxygenase-2, representing that inhibition occurs at the transcriptional level. In addition, we performed molecular docking and molecular dynamic simulations to study the interaction of 5-hydroxy-3′,4′,7-trimethoxyflavone with inflammatory mediators such as inducible nitric oxide synthase and cyclooxygenase-2. Docking study showed its hydrogen bond interactions with key residues in the active site of inducible nitric oxide synthase and cyclooxygenase-2, enlightening its possible binding mode at the molecular level. The results of molecular dynamic simulations showed the stability of complexes and their interactions. Taken together, these findings envisage 5-hydroxy-3′,4′,7-trimethoxyflavone as a potential candidate molecule for the progress of therapeutic strategy against inflammation-related diseases.

Nanosilver crystals capped with Bauhinia acuminata phytochemicals as new antimicrobials and mosquito larvicides

To develop novel nanoformulated insecticides and antimicrobials, herein we produced Ag nanoparticles (AgNPs) using the Bauhinia acuminata leaf extract. This unexpensive aqueous extract acted as a capping and reducing agent for the formation of AgNPs. We characterized B. acuminata-synthesized AgNPs by UV-vis and FTIR spectroscopy, XRD and TEM analyses. UV-vis spectroscopy analysis of B. acuminata-synthesized AgNPs showed a peak at 441.5 nm. FTIR shed light on functional groups from the phytoconstituents involved in nanosynthesis. XRD of B. acuminata-synthesized AgNPs suggested a face-centered cubic structure, with a highly crystalline nature. TEM of B. acuminata-synthesized AgNPs revealed mean size of 25 nm, with round shape. AgNPs tested at 60 μg/mL inhibited the growth of 5 bacteria and 3 fungal pathogens. In the insecticidal assays on important mosquito species, LC50 of the aqueous extract of B. acuminata leaves on the larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus were 204.07, 226.02, and 249.24 μg/mL, respectively. The B. acuminata-synthesized AgNPs exhibited higher larvicidal efficacy, with LC50 values of 24.59, 27.19, and 30.19 μg/mL, respectively. Therefore, herein we developed a single-step, reliable, inexpensive, and environmentally non-toxic synthesis process to obtain AgNPs with high bioactivity against pathogens and vectors. Given the effective antimicrobial and larvicidal activity, nanoparticles fabricated using plant extracts and extremely low concentrations of trace elements, such as silver, can be exploited for multipurpose activities. Our results pointed out that B. acuminata-synthesized AgNPs have a promising potential in antimicrobial food packaging, as well as a foliar spray to control plant pathogens in the field, and to synergize the efficacy of fungicidal and larvicidal formulations.

Antiproliferative and apoptosis-induction studies of 5-hydroxy 3′,4′,7-trimethoxyflavone in human breast cancer cells MCF-7: an in vitro and in silico approach

Abstract The aim of this study was to find the efficacy of 5-hydroxy 3′,4′,7-trimethoxyflavone (HTMF), a flavonoid compound isolated from the medicinal plant Lippia nodiflora, in inhibiting the proliferation and inducing apoptosis in human breast cancer cell line MCF-7. The anti-proliferative effect of the compound HTMF was confirmed using MTT cytotoxicity assay. Increased apoptotic induction by HTMF was demonstrated by acridine orange/ethidium bromide (AO/EtBr) and Hoechst 33258 staining studies. The phosphatidylserine translocation, an early feature of apoptosis and DNA damage were revealed through AnnexinV-Cy3 staining and comet assay. Moreover, the significant elevation of cellular ROS was observed in the treated cells, as measured by 2,7-diacetyl dichlorofluorescein (DCFH-DA). The mRNA expression studies also supported the effectiveness of HTMF by shifting the Bax:Bcl-2 ratio. The treatment of MCF-7 cells with HTMF encouraged apoptosis through the modulation of apoptotic markers, such as p53, Bcl-2, Bax, and cleaved PARP. In silico molecular docking and dynamics studies with MDM2-p53 protein revealed that HTMF was more potent compound that could inhibit the binding of MDM2 with p53 and, therefore, could trigger apoptosis in cancer cell. Overall, this study brings up scientific evidence for the efficacy of HTMF against MCF-7 breast cancer cells. Graphical Abstract