INHIBITION OF ADVANCED GLYCATION END-PRODUCT FORMATION BY QUERCETIN AND CATECHIN: AN ALTERNATIVE THERAPY FOR TREATING DIABETIC COMPLICATIONS

  Objective: The objective of this research was to determine early advanced glycation end-product (AGE) inhibition by natural aldose reductase inhibitors (ARIs), quercetin and catechin.Methods: The assay mixture (4 ml) consisted of 2 ml of 50 mM phosphate-buffered saline (pH 7.4), 50 μg/μl bovine serum albumin (BSA), and 2 mM glucose with or without the inhibitor. The test samples were treated with three different concentrations (10 mM, 20 mM, and 40 mM) of quercetin and catechin. High-throughput screening-based assay was adapted to perform the BSA-glucose test to determine the induction of AGE formation and its inhibition by quercetin, and catechin, using the fluorescence of the AGE-BSA sample at excitation and emission wavelengths of 350 and 450 nm.Result: The ARIs, quercetin and catechin inhibited early glycation with an inhibitory concentration value of 15.58 mM and 35.01 mM, respectively.Conclusion: The suppression of AGEs formation by natural inhibitors of aldose reductase would provide an alternative approach to the control of diabetic complications

EFFECT OF TOLL-LIKE RECEPTOR INHIBITOR IMIQUIMOD ON IL1R1 INTERACTION WITH IL-1RA AND ITS SNP VARIANT-AN IN SILICO APPROACH

Objective: Interleukin 1 receptor antagonist (IL1Ra) acts as an antagonist to Interleukin 1 beta (IL1β) signalling in maintaining homeostasis. A loss of function due to Single Nucleotide Polymorphism (SNP) occurrence in IL1Ra can lead to dysregulated state as seen in autoimmune disease pathogenesis. The current study aims at achieving conformational stability in the IL1R1-IL1Ra_SNP complex by introducing a ligand into the region apart from the active site of Interleukin 1 receptor type1 (IL1R1).Methods: Protein-protein docking was performed using ClusPro, for IL1R1 with IL1β, IL1Ra and IL1Ra_SNP variant to study the difference in the interaction between the complexes. A known inhibitor, Imiquimod was docked using Glide, to the Il1R1-IL1Ra_SNP complex using flexible docking and the change in surface energy was calculated.Results: Binding Interactions show that IL1Ra binds more avidly to IL1R1 than IL1β. Conformational instability was seen in the IL1R1-IL1Ra_SNP complex. The difference in the amino acid interactions between the IL1R1 with IL1Ra and IL1Ra_SNP variant further illustrated the change in binding residues and hydrogen bond formation. Upon docking of an appropriate ligand to the IL1R1-IL1Ra_SNP complex, the conformational stability of the IL1R1-IL1Ra_SNP complex enhanced considerably suggesting a possible mechanism for treating SNP-induced conformational instability.Conclusion: Toll-like receptors like IL1R1 have many binding pockets apart from its active site. No strategies have yet been reported in targeting them for correcting conformational instability induced by SNP. Through our study, it was observed that the conformational instability of IL1R1-IL1Ra_SNP complex decreased upon introducing an appropriate small molecule.Keywords: IL1β, IL1R1, IL1Ra, SN

COMPUTATIONAL ANALYSIS OF INTERACTIONS BETWEEN ANTI-EPILEPTIC DRUGS AND IMPORTANT PLACENTAL PROTEINS-A POSSIBLE ROUTE FOR NEURAL TUBE DEFECTS IN HUMANS

Objective: The reason behind the occurrence of Neural Tube Defects (NTDs) in pregnant women treated with certain Anti-Epileptic Drugs (AEDs) such as Carbamazepine, Valproate, Lamotrigine, Phenobarbital, etc., is not yet known. The relationship between Folic Acid intake and NTDs is not yet established. Folate receptors play a critical role in mediating placental transport of maternal folate to the foetus. Another important protein is Carnitine O-acetyltransferase that is involved in the transport of carnitine, which is much needed for foetal metabolic functions and tissue development. The objective of this study is to understand the interaction of AEDs with two important placental proteins through a docking approach to establishing a suitable explanation of AED's role in NTD.Methods: A generic algorithm based docking was used to identify and study the mode of interactions between the drugs and placental proteins. For comparison purpose, the natural ligands of these receptors have also been included in the dataset containing AEDs.Results: Both bonded and non-bonded interactions were observed between AEDs and the crucial residues of these proteins. The drugs formed complex with these proteins with satisfactory binding energy. Some amount of Electrostatic interaction was also observed among a few pairs of protein residue and drug molecules.Conclusion: We suggest that these drug-protein associations, involving bonded and non-bonded interactions, could be a possible portal by which certain AEDs induce NTDs in the foetus. Higher interaction of Pantothenic Acid with Folate Receptor could be a mechanism through which Pantothenic Acid inhibits Valproic Acid-induced NTDs. And thus, its supplementation can specifically prevent Valproic Acid-induced NTDs. The above mechanism also explains how increased intake of Folic Acid during pregnancy can reduce the occurrence of NTDs.Keywords: Neural Tube Defect, Anti-epileptic drug, Placenta, Folate transporters, Docking, Carnitine O-acetyltransferas

COMPUTATIONAL ANALYSIS OF COMPOUNDS FROM OCIMUM SANCTUM FOR ANTICANCER ACTIVITY AGAINST ORAL SQUAMOUS CELL CARCINOMA

Objective: The objectives of this research are to identify the potentials of active phytochemicals from Ocimum sanctum as anticancer agents, by inhibiting the epidermal growth factor receptor (EGFR), one of the highly expressed proteins inducing metastasis in oral squamous cell carcinoma (OSCC) as well as other cancers.Methods: The phytochemicals found in O. sanctum were identified and downloaded from online chemical databases. The target protein was retrieved from the Protein Data Bank. Virtual screening using glide protocols of high throughput virtual screening and molecular docking using standard precision and extra precision (XP) were carried out. The binding energies and the important physicochemical properties of the compounds were also determined.Results: A total number of 210 compounds from O. sanctum were screened against EGFR. Lipinski rule was followed to find the compounds with favorable drug absorptive properties. The shortlisted compounds, namely luteolin, apigenin, and isothymusin, possess high Glide scores (kcal/mol) of −9.98, −9.51, and −9.45 and binding energies (kcal/mol) of −42.63, −48.28, and −44.95, respectively.Conclusion: Among the three compounds, Isothymusin was not yet been reported to posess anticancer activity. Our study suggest this compound as a potential chemotherapeutic agent for treating OSCC. They function by inhibiting the activity of metastasis - inducing protein EGFR

SYNTHESIS, CRYSTAL STUDIES AND PHARMACOLOGICAL ROLE PREDICTION OF 3-IODO-2- METHYL-1 PHENYL SULFONYL-1H INDOLE

ABSTRACTObjective: Indole-based compounds have established many pharmacological applications. Indole acts as a starting compound for various medicinalpreparations. Our objective is to determine the possible pharmacological roles of our crystallized indole-based ligand using computational approach.The activities studied are antibacterial, antitubercular, and antimelanoma.Methods: The structure of the indole compound, CR2 was studied using single crystal X-ray diffraction technique. To predict the pharmacologicalactivities, the structure-based docking method was followed. The protein targets were selected based on their important biological role in eachactivity mentioned above. The interactions between the targets and the crystal ligand were studied using a generic based docking algorithm. Thesignificant pharmacophore features of the ligand were also reported.Results: The ligand showed bonded and non-bonded interactions with the crucial amino acid residues of the active site of each target. The interactionsand the binding energies were quite comparable to the targets' natural ligands.Conclusion: We suggest through the computational approaches that the ligand may have antitubercular, antibacterial, and antimelanoma activitieswith respect to the targets considered. The new insights of this compound as predicted by the computational methods are believed to provide aplatform for the futuristic pharmacological activities of this compound that can be further considered for wet lab techniques.Keywords: Indole, Dihydrofolate reductase, DNA gyrase, Human B-Raf kinase, Docking, Pharmacophore, Single crystal X-ray diffraction

ANALYSIS OF MOLECULAR DOCKING EFFICIENCY OF CLEISTANTHIN-A, AS AN ALTERNATIVE FOR NICOTINE ADDICTION

Objective: The present research was aimed to understand the molecular docking efficiency of a plant-derived compound cleistanthin-A and a common ingredient in tobacco consumption nicotine with nicotinic acetylcholine receptor (nAChR).Methods: The 3-D structure of nAChR was retrieved from the protein data bank (ID 5AFH). Ligand was obtained from the PUBCHEM. The in silico protocol comprised of three steps: high-throughput virtual screening (HTVS), standard preci­sion (SP) and extra precision (XP). The screened molecules were ranked accordingly using glide score. Schrödinger tool was used to perform the docking analysis.Results: The binding efficiency of the nicotine and cleistanthin-A was found to be docked at the cys-cys loop of the receptor. Based upon the glide score and glide energy it can be reported that, nicotine binding can be inhibited by the binding of cleistanthin-A to the nAChR.Conclusion: The docking efficiency of cleistanthin-A was good compared to nicotine towards nAChR. Hence, cleistanthin–A was derived as a better choice as an alternative for nicotine in smoke therapy

Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria

<p>Abstract</p> <p>Background</p> <p>The six organic solvent extracts of <it>Artemisia nilagirica </it>were screened for the potential antimicrobial activity against phytopathogens and clinically important standard reference bacterial strains.</p> <p>Methods</p> <p>The agar disk diffusion method was used to study the antibacterial activity of <it>A. nilagirica </it>extracts against 15 bacterial strains. The Minimum Inhibitory Concentration (MIC) of the plant extracts were tested using two fold agar dilution method at concentrations ranging from 32 to 512 μg/ml. The phytochemical screening of extracts was carried out for major phytochemical derivatives in <it>A. nilagirica</it>.</p> <p>Results</p> <p>All the extracts showed inhibitory activity for gram-positive and gram-negative bacteria except for <it>Klebsiella pneumoniae, Enterococcus faecalis </it>and <it>Staphylococcus aureus</it>. The hexane extract was found to be effective against all phytopathogens with low MIC of 32 μg/ml and the methanol extract exhibited a higher inhibition activity against <it>Escherichia coli, Yersinia enterocolitica, Salmonella typhi</it>, <it>Enterobacter aerogenes</it>, <it>Proteus vulgaris</it>, <it>Pseudomonas aeruginosa </it>(32 μg/ml), <it>Bacillus subtilis </it>(64 μg/ml) and <it>Shigella flaxneri </it>(128 μg/ml). The phytochemical screening of extracts answered for the major derivative of alkaloids, amino acids, flavonoids, phenol, quinines, tannins and terpenoids.</p> <p>Conclusion</p> <p>All the extracts showed antibacterial activity against the tested strains. Of all, methanol and hexane extracts showed high inhibition against clinical and phytopathogens, respectively. The results also indicate the presence of major phytochemical derivatives in the <it>A. nilagirica </it>extracts. Hence, the isolation and purification of therapeutic potential compounds from <it>A. nilagirica </it>could be used as an effective source against bacterial diseases in human and plants.</p

ENERGY-BASED PHARMACOPHORE MODELING, VIRTUAL SCREENING, AND MOLECULAR DYNAMICS TO IDENTIFY POTENTIAL INHIBITORS FOR GLYCOGEN SYNTHASE KINASE 3 BETA

  Objective: Glycogen synthase kinase 3 beta (GSK3β) is one of the main targets for wound healing activity. Our objective is to identify novel inhibitors for GSK3β using in silico approach.Methods: Grid-based molecular docking, energy-based pharmacophore (e-pharmacophore) modeling, and molecular dynamics (MD) studies were performed for phytocompounds with GSK3β and compared with standard drugs using Schrodinger software.Results: The glide scores and the molecular interactions of the phytocompounds were well comparable to the standard drugs. The MD was performed for the target bound to the best scoring ligand, entagenic acid. The pharmacophore features of this docked complex were modeled as e-pharmacophore. The constructed e-pharmacophore model was screened against phytocompounds retrieved from literature to identify the ligands with similar pharmacophore features.Conclusion: The glide scores of fukinolic acid, cimicifugic acid, and linarin were −10.99, −8.28, and −7.25 kcal/mol, respectively. The further 50 nanoseconds MD study determined the stability of GSK3β-linarin complex. Nitrofurazone and sulfathiazole drugs can lead to systemic side effects. Hence, it is concluded that linarin could be a potent wound healing compound against GSK3β

INHIBITION OF ADVANCED GLYCATION END-PRODUCT FORMATION BY QUERCETIN AND CATECHIN: AN ALTERNATIVE THERAPY FOR TREATING DIABETIC COMPLICATIONS

  Objective: The objective of this research was to determine early advanced glycation end-product (AGE) inhibition by natural aldose reductase inhibitors (ARIs), quercetin and catechin.Methods: The assay mixture (4 ml) consisted of 2 ml of 50 mM phosphate-buffered saline (pH 7.4), 50 μg/μl bovine serum albumin (BSA), and 2 mM glucose with or without the inhibitor. The test samples were treated with three different concentrations (10 mM, 20 mM, and 40 mM) of quercetin and catechin. High-throughput screening-based assay was adapted to perform the BSA-glucose test to determine the induction of AGE formation and its inhibition by quercetin, and catechin, using the fluorescence of the AGE-BSA sample at excitation and emission wavelengths of 350 and 450 nm.Result: The ARIs, quercetin and catechin inhibited early glycation with an inhibitory concentration value of 15.58 mM and 35.01 mM, respectively.Conclusion: The suppression of AGEs formation by natural inhibitors of aldose reductase would provide an alternative approach to the control of diabetic complications

ENERGY-BASED PHARMACOPHORE MODELING, VIRTUAL SCREENING, AND MOLECULAR DYNAMICS TO IDENTIFY POTENTIAL INHIBITORS FOR GLYCOGEN SYNTHASE KINASE 3 BETA

  Objective: Glycogen synthase kinase 3 beta (GSK3β) is one of the main targets for wound healing activity. Our objective is to identify novel inhibitors for GSK3β using in silico approach.Methods: Grid-based molecular docking, energy-based pharmacophore (e-pharmacophore) modeling, and molecular dynamics (MD) studies were performed for phytocompounds with GSK3β and compared with standard drugs using Schrodinger software.Results: The glide scores and the molecular interactions of the phytocompounds were well comparable to the standard drugs. The MD was performed for the target bound to the best scoring ligand, entagenic acid. The pharmacophore features of this docked complex were modeled as e-pharmacophore. The constructed e-pharmacophore model was screened against phytocompounds retrieved from literature to identify the ligands with similar pharmacophore features.Conclusion: The glide scores of fukinolic acid, cimicifugic acid, and linarin were −10.99, −8.28, and −7.25 kcal/mol, respectively. The further 50 nanoseconds MD study determined the stability of GSK3β-linarin complex. Nitrofurazone and sulfathiazole drugs can lead to systemic side effects. Hence, it is concluded that linarin could be a potent wound healing compound against GSK3β