ABSTRACT
In a sequel to investigate osteogenic potential of ethanolic extract of Cissus quadrangularis (CQ), the present study reports the osteoblast differentiation and mineralization potential of ethyl acetate (CQ-EA) and butanol (CQ-B) extracts of CQ on mouse pre-osteoblast cell line MC3T3-E1 (sub-clone 4) with an objective to isolate an antiosteoporotic compound. Growth curve, proliferation, and viability assays showed that both the extracts were nontoxic to the cells even at high concentration (100 µg/ml). The cell proliferation was enhanced at low concentrations (0.1 µg/ml and 1 µg/ml) of both the extracts. They also upregulated the osteoblast differentiation and mineralization processes in MC3T3-E1 cells as reflected by expression profile of osteoblast marker genes such as RUNX2, Osterix, Collagen (COL1A1), Alkaline Phosphatase (ALP), Integrin-related Bone Sialoprotein (IBSP), Osteopontin (OPN), and Osteocalcin (OCN). CQ-EA treatment resulted in early differentiation and mineralization as compared with the CQ-B treatment. These findings suggest that low concentrations of CQ-EA and CQ-B have proliferative and osteogenic properties. CQ-EA, however, is more potent osteogenic than CQ-B.
Subject(s)
Calcification, Physiologic/drug effects , Cissus/chemistry , Osteoblasts/drug effects , Plant Extracts/pharmacology , 1-Butanol/chemistry , Acetates/chemistry , Alkaline Phosphatase/metabolism , Animals , Bone Morphogenetic Protein 2/metabolism , Cell Differentiation/drug effects , Cell Line , Cell Proliferation/drug effects , Mice , Osteoblasts/metabolism , Osteocalcin/metabolism , Osteogenesis/drug effects , Osteopontin/metabolism , Plant Extracts/chemistry , Up-Regulation/drug effectsABSTRACT
Virtual screening uses computer based methods to discover new ligands on the basis of biological structures. Among all virtual screening methods structure based docking has received considerable attention. In an attempt to identify new ligands as urease inhibitors, structure-based virtual screening (SBVS) of an in-house database of 10,000 organic compounds was carried out. The X-ray crystallographic structure of Bacillus pasteurii (BP) in complex with acetohydroxamic acid (PDB Code 4UBP) was used as a protein structure. As a starting point, ~10,000 compounds of our in-house database were analyzed to check redundancy and the compounds found repeated were removed from the database. Finally 6993 compounds were docked into the active site of BP urease using GOLD and MOE-Dock software. A remarkable feature of this study was the identification of monastrol, a well-known KSP inhibitor already in clinical trials, as a novel urease inhibitor. The hits identified were further evaluated by molecular docking and on examination of the affinity predictions, twenty-seven analogs of monastrol were synthesized by a multicomponent Biginelli reaction followed by their in vitro screening as urease inhibitors. Finally twelve compounds were identified as new urease inhibitors. The excellent in vitro activity suggested that these compounds may serve as viable lead compounds for the treatment of urease related problems.
Subject(s)
Pyrimidines/metabolism , Thiones/metabolism , Urease/antagonists & inhibitors , Urease/ultrastructure , Bacillus/enzymology , Catalytic Domain , Crystallography, X-Ray , Drug Design , Drug Evaluation, Preclinical , Hydroxamic Acids/chemistry , Hydroxamic Acids/metabolism , Kinesins/antagonists & inhibitors , Ligands , Models, Molecular , Molecular Docking SimulationABSTRACT
Elaeagnoside (1), a new steroidal glucoside, has been isolated from the chloroform fraction of Elaeagnus orientalis along with beta-sitosterol and 12-hydroxy-8,10-octadecadienoic acid. The structure of 1 has been elucidated with the help of chemical and spectral studies. It showed significant inhibitory activity against the enzyme chymotrypsin.
