Isonicotinohydrazones as inhibitors of alkaline phosphatase and ecto‐5′‐nucleotidase

Abstract

A series of isonicotinohydrazide derivatives was synthesized and tested against recombinant human and rat ecto-5'-nucleotidases (h-e5'NT and r-e5'NT) and alkaline phosphatase isozymes including both bovine tissue-non-specific alkaline phosphatase (b-TNAP) and tissue-specific calf intestinal alkaline phosphatase (c-IAP). These enzymes are implicated in vascular calcifications, hypophosphatasia, solid tumors, and cancers, such as colon, lung, breast, pancreas, and ovary. All tested compounds were active against both enzymes. The most potent inhibitor of h-e5'NT was derivative (E)-N'-(1-(3-(4-fluorophenyl)-5-phenyl-4,5-dihydro-1H-pyrazol-1-yl)ethylidene)isonicotinohydrazide (3j), whereas derivative (E)-N'-(4-hydroxy-3-methoxybenzylidene)isonicotinohydrazide (3g) exhibited significant inhibitory activity against r-e5'NT. In addition, the derivative (E)-N'-(4'-chlorobenzylidene)isonicotinohydrazide (3a) was most potent inhibitor against calf intestinal alkaline phosphatase and the derivative (E)-N'-(4-hydroxy-3-methoxybenzylidene)isonicotinohydrazide (3g) was found to be most potent inhibitor of bovine tissue-non-specific alkaline phosphatase. Furthermore, putative binding modes of potent compounds against e5'NT (human and rat e5'NT) and AP (including b-TNAP and c-IAP) were determined computationally.

Synthesis of novel ( E )-1-(2-(2-(4(dimethylamino) benzylidene) hydrazinyl)-4-methylthiazol-5-yl)ethanone derivatives as ecto-5′-nucleotidase inhibitors

“…Molecular docking analysis of every compound was performed to explore binding mode with the active site of h -e5′NT as well as r -e5′NT target enzymes. X-ray crystallographic structures were present in RCSB Protein Data Bank which was downloaded in the form of PDB ID: 4H2I [ 33 ], while X-ray crystallographic structure of r -e5′NT was not reported till now, so a previous homology model was selected for docking studies [ 34 ]. The two-dimensional structures of all compounds T-1 to T-6 were created via Marvin-ChemAxon suit [ 35 ] and transformed into a three-dimensional structure with the Molecular Builder program executed in Molecular Operating Environment (MOE 2014.0901) [ 36 ].…”

Ectonucleotidase Inhibitory and Redox Activity of Imidazole‐Based Organic Salts and Ionic Liquids

“…Ecto‐5′‐nucleotidase inhibition assay : The enzyme inhibition assay against h ‐e5′NT and r ‐e5′NT was performed according to a previously described method . In this method, the P/ACE MDQ capillary electrophoresis system (Beckman Instruments, Fullerton, CA, USA) was used, which used a UV detection system for evaluation.…”

“…[39] Ecto-5'-nucleotidase inhibition assay:T he enzyme inhibition assay against h-e5'NT and r-e5'NT was performed according to a previously described method. [40] In this method, the P/ACE MDQ capillary electrophoresis system (Beckman Instruments, Fullerton, CA, USA) was used, which used aU Vd etection system for evaluation. The selected compounds were prepared in DMSO stock solution (10 %, 10 mm)a nd further diluted in assay buffer (10 mm Tris·HCl buffer of pH 7.4 containing 1mm CaCl 2 and 2mm MgCl 2 ) to obtain working solutions (1 mm).…”

Domino Reactions of Chromone‐3‐carboxylic Acids with Aminoheterocycles: Synthesis of Heteroannulated Pyrido[2,3‐c]coumarins and their Optical and Biological Activity

“…Cell Transfection with e5′NT: The cell transfection of e5′NT was carried out according to a previously reported method . For this, COS‐7 cells were transfected with the plasmids expressing e5′NT (human and rat) in 10 cm plates using Lipofectamine. The confluent cells (80–90 %) were incubated at 37 °C for 5 h in Dulbecco's modified Eagle medium (DMEM) [without fetal bovine serum (FBS)] with plasmid DNA (6 µg) and Lipofectamine reagent (24 µL).…”