Synthesis and characterization of novel 1,2-oxazine-based small molecules that targets acetylcholinesterase

Thirteen 2-oxazine-based small molecules were synthesized targeting 5-lipoxygenase (LOX), and acetylcholinesterase (AChE). The test revealed that the newly synthesized compounds had potent inhibition towards both 5-LOX and AChE in lower micro molar concentration. Among the tested compounds, the most active compound, 2-(2-acetyl-6,6-dimethyl-4-phenyl-5,6-dihydro-2H-1,2-oxazin-3- yl)methyl-1H-isoindole-1,3(2H)-dione (2a) showed inhibitory activity towards 5-LOX and AChE with an IC50 values of 1.88, and 2.5 μM, respectively. Further, the in silico molecular docking studies revealed that the compound 2a bound to the catalytic domain of AChE strongly with a highest CDOCKER score of -1.18 kcal/mol when compared to other compounds of the same series. Additionally, 2a showed a good lipophilicity (log P = 2.66), suggesting a potential ability to penetrate the blood-brain-barrier. These initial pharmacological data revealed that the compound 2a could serve as a drug-seed in developing anti-Alzheimer's agents. © 2014 Elsevier Ltd. All rights reserved

Anti-tumor and anti-angiogenic activity of novel hydantoin derivatives: Inhibition of VEGF secretion in liver metastatic osteosarcoma cells

A series of new azaspiro bicyclic hydantoin derivatives has been designed and synthesized. Initially, the anti-proliferative effect of the hydantoin derivatives was evaluated against human ovarian cancer cells (SKOV-3 and OVSAHO) and murine osteosarcoma cells (LM8 and LM8G7). Among the tested compounds, 8-(3-fluorobenzyl)-1′-(4-(methylsulfonyl)benzyl)-8-azaspiro[bicyclo[3.2.1]octane-3,4′-imidazolidine]-2′,5′-dione (7h) and 8-(3,4-difluorobenzyl)-1′-(4-(methylsulfonyl)benzyl)-8-azaspiro[bicyclo[3.2.1]octane-3,4′-imidazolidine]-2′,5′-dione (7i) showed a significant anti-proliferative activity against the OVSAHO and LM8G7 cells. The real-time monitoring of the effect of the compounds 7h and 7i against the proliferation of LM8G7 was revealed that resulting IC50 values were 102 μM and 13 μM, respectively. We reasoned that the presence of fluorine atom at the 3rd position of the phenyl ring of the hydantoin side chain may determine the potency of the molecule. Furthermore, the compound 7i inhibited the tube formation of the mouse endothelial cells. Finally, the treatment of the compound 7i against the proliferation of LM8G7 cells demonstrated the down regulation of the secretion of VEGF, indicate the potential angioinhibitory effects. In conclusion, our findings demonstrate the suppression of the secretion of VEGF by LM8G7 cells by the compound 7i might contribute at least in part to the antitumor action

PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF L-AMINO ACID OXIDASE FROM WESTERN REGION INDIAN COBRA (NAJA NAJA) VENOM

Objective: Purification and biochemical characterization of LAAO from western region Indian Cobra (Naja naja) venom.Methods: LAAO was purified from Indian cobra (Naja naja) venom using sequential chromatography on Sephadex G-75 gel filtration followed by Ion exchange on CM-Sephadex C-25 column. Biochemical characterization viz., pH, Temperature, Km and Vmax were determined. Molecular weight of LAAO was determined by electrophoresis. Inhibition of LAAO from cold water extracts of Curcuma zedoria, Curcuma ceasia, Curcuma aromatic, Curcuma longa, Curcuma amada, Cucumis sativus and Benincasa hispida was done.Results: Purified LAAO showed the single band on non reducing SDS-PAGE with approximate molecular weight of 65 kDa. Further biochemical characterization revealed that, LAAO from Naja naja (western region) has an optimum pH of 7.0 and is stable at room Temperature upto 37 °C and showed an optimum enzyme concentration of 4µg/ml and Km 134.1µM and Vmax is 21.87 U/min. Cold water extract of Curcuma ceasia, Curcuma aromatic and Benincasa hispida showed comparatively significant inhibition of LAAO.Conclusion: LAAO has promising therapeutic prospects because of its effects on various biological functions. Variation in snake species and their geographical distribution also contributes to the venom properties such as composition, toxicity level, pharmacological and biological activities. The significant difference in protein profiling leads to substantial lethality among different geographical regions. Thus in this study Indian Cobra (Naja naja) venom from the western region of India was subjected to purification of LAAO and biochemical characterization. Â

Serine protease from Indian Cobra venom: its anticoagulant property and effect on human fibrinogen

An unusual low molecular weight serine protease (NnP28) has been purified from Indian Cobra (Naja naja) venom from the western region of the Indian sub-continent. We reported the purification and characterization of low molecular NnP28, emphasizing its role on human fibrinogen and anticoagulant property. NnP28 was purified using gel filtration column chromatography followed by ion exchange chromatography. Protein gel electrophoresis revealed its molecular weight approximate to 28 kDa. The exact molecular mass of NnP28 was found to be 27.12 kDa by mass spectrometry, hydrolyzing casein specifically, inhibited by PMSF suggesting it has a serine protease. NnP28 prolonged the clotting time of re-calcified human citrated plasma and activated partial thromboplastin time (APTT) exhibiting anticoagulant property. NnP28 exhibited fibrinogenolytic activity. Thus, the present study demonstrates the presence of unusual low molecular weight serine protease, emphasizing its importance of region-specific Indian cobra species

1-(3-Bromo-2-thien­yl)ethanone

In the title compound, C6H5BrOS, the non-H and aromatic H atoms lie on a crystallographic mirror plane. In the crystal, mol­ecules are linked into chains propagating along the c axis by inter­molecular C—H⋯O hydrogen bonds

N-[4-Cyano-3-(trifluoro­meth­yl)phen­yl]-2-meth­oxy­benzamide

In the title compound, C16H11F3N2O2, the carboxamide group connecting the two aromatic rings is in a syn-periplanar configuration; the mol­ecule is non-planar; the dihedral angle between the two aromatic rings is 13.95 (18)°. Intra­molecular N—H⋯O and C—H⋯O hydrogen bonds occur. In the crystal, mol­ecules are linked by inter­molecular C—H⋯O hydrogen bonds

Plumbagin inhibits invasion and migration of breast and gastric cancer cells by downregulating the expression of chemokine receptor CXCR4

<p>Abstract</p> <p>Background</p> <p>Increasing evidence indicates that the interaction between the CXC chemokine receptor-4 (CXCR4) and its ligand CXCL12 is critical in the process of metastasis that accounts for more than 90% of cancer-related deaths. Thus, novel agents that can downregulate the CXCR4/CXCL12 axis have therapeutic potential in inhibiting cancer metastasis.</p> <p>Methods</p> <p>In this report, we investigated the potential of an agent, plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone), for its ability to modulate CXCR4 expression and function in various tumor cells using Western blot analysis, DNA binding assay, transient transfection, real time PCR analysis, chromatin immunoprecipitation, and cellular migration and invasion assays.</p> <p>Results</p> <p>We found that plumbagin downregulated the expression of CXCR4 in breast cancer cells irrespective of their HER2 status. The decrease in CXCR4 expression induced by plumbagin was not cell type-specific as the inhibition also occurred in gastric, lung, renal, oral, and hepatocellular tumor cell lines. Neither proteasome inhibition nor lysosomal stabilization had any effect on plumbagin-induced decrease in CXCR4 expression. Detailed study of the underlying molecular mechanism(s) revealed that the regulation of the downregulation of CXCR4 was at the transcriptional level, as indicated by downregulation of mRNA expression, inhibition of NF-κB activation, and suppression of chromatin immunoprecipitation activity. In addition, using a virtual, predictive, functional proteomics-based tumor pathway platform, we tested the hypothesis that NF-κB inhibition by plumbagin causes the decrease in CXCR4 and other metastatic genes. Suppression of CXCR4 expression by plumbagin was found to correlate with the inhibition of CXCL12-induced migration and invasion of both breast and gastric cancer cells.</p> <p>Conclusions</p> <p>Overall, our results indicate, for the first time, that plumbagin is a novel blocker of CXCR4 expression and thus has the potential to suppress metastasis of cancer.</p

Synthesis and characterization of novel 6-fluoro-4-piperidinyl-1,2-benzisoxazole amides and 6-fluoro-chroman-2-carboxamides: antimicrobial studies

Novel derivatives of 6-fluoro-4-piperidinyl-1,2-benzisoxazole amides 4(I–VI) were obtained by the condensation of different acid chlorides with 6-fluoro-3-piperidin-4yl-benzodisoxazole. Also, 6-fluoro-chroman-2-carboxamides 6(I–III) were synthesized by using nebulic acid chloride with different amines in presence of triethylamine as acid scavenger and dichloroethane as solvent. The synthesized compounds were characterized by IR, 1H NMR, and CHN analysis. These molecules were evaluated for their efficacy as antimicrobials in vitro by disc diffusion and microdilution method against pathogenic strains such as Bacillus substilis, Escherichia coli, Pseudomonas fluorescens, Xanthomonas campestris pvs, X. oryzae, Aspergillus niger, A. flavus, Fusarium oxysporum, Trichoderma species, F. monaliforme, and Penicillum species. Compounds 4I, 4IV, 4V, 6I, 6II and 6III showed better inhibitory activity than compared to standard drugs. Among these compounds, 4IV and 6III showed potent inhibitory activity against all the strains and found to be nonstrain dependent. The title compounds represent a novel class of potent antimicrobial agents

Synthesis and crystal structure of 1-(2-Bromo-4,5-dimethoxybenzyl)-benzod1,2,3triazole

The title compound C15H14N3O2Br, was synthesized and characterized by X-ray diffraction analysis. The compound crystallizes in triclinic crystal class in the space group P1. The cell parameters are a = 7.057(1)Å, b = 8.142(1)Å, c = 13.738(2)Å, α = 90.039(7)°, β = 96.869(5)°, γ = 108.047(8)° and Z = 2. The final residual factor R1 = 0.0513