Carbonic anhydrases (CAs) are known as a drug-target enzymes. The inhibitors of the enzyme are important compounds for discovering new therapeutic agents and understanding in detail protein-drug interactions at the molecular level. For this purpose, the in vitro effects of some anti-inflammatory agents such as tenoxicam, fluorometholone acetate, and dexamethasone were investigated on esterase activity of human erythrocyte CA-I and CA-II in this study. hCA-I and hCA-II were purified by affinity chromatography with a yield of 47.25% and 87%, and a specific activity of 642.8 EU/mg proteins and 5576.9 EU/mg proteins, respectively. SDS-PAGE was performed to determine the purity of the enzymes. Inhibitory effects of the drugs on hCA-I and hCA-II were determined by spectrophotometric method. IC50 values for hCA-I and hCA-II were 0.198, 2.18, 11.7, 0.11, 17.5 and 14 μm using tenoxicam, fluorometholone acetate, and dexamethasone, respectively. For fluorometholone acetate and dexamethasone, Ki values from Lineweaver-Burk plots were obtained as 1.044 and 21.2 μm (noncompetitive) for hCA-I and 9.98 and 8.66 μm (non-competitive) for hCA-II. In conclusion, tenoxicam, fluorometholone acetate, and dexamethasone showed potent inhibitory effects on esterase activity of hCA-I and hCA-II isozymes under in vitro conditions.
In this study a series of pyrazole-3,4-dicarboxamide (3-10) derivatives bearing sulfonamide moiety were synthesized starting from 1-(3-nitrophenyl)-5-phenyl-1H-pyrazole-3,4-dicarboxylic acid (1). The structures of synthesized molecules were characterized by FT-IR, 1 H NMR, 13 C NMR, and elemental analysis methods. Human carbonic anhydrase isoenzymes (hCA I and hCA II) were purified separately from erythrocyte cells by the Sepharose-4B-L-tyrosine-sulfa nilamide affinity column chromatography and inhibitory effects of newly synthesized sulfonamides on esterase activities of these isoenzymes have been studied as in vitro. The K i values of compounds were found in the range of 0.056-110.400 lM for hCA I and 0.057-533.400 lM for hCA II. Compound 4 has the highest inhibitory effect for hCA I and hCA II while compound 5 showed lowest inhibition. The structure-activity relationships for the inhibition of these isoforms with the pyrazole-sulfonamides reported here were also elucidated. ª 2015 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Background:
Persistent oxidative stress can lead to chronic inflammation and
mediate most chronic diseases including neurological disorders. Oleuropein has been
shown to be a potent antioxidant molecule in olive oil leaf having antioxidative properties.
Objective:
The aim of this study was to investigate the protective effects of oleuropein
against oxidative stress in human glioblastoma cells.
Methods:
Human glioblastoma cells (U87) were pretreated with oleuropein (OP) essential
oil 10 µM. After 30 minutes, 100 µM H2O2 was added to the cells for three hours. Cell
survival was quantified by colorimetric MTT assay. Glutathione level, total oxidant capacity,
total antioxidant capacity and nitric oxide levels were determined by using specific
spectrophotometric methods. The relative gene expression level of iNOS was performed by
qRT-PCR method.
Results:
According to viability results, the effective concentration of H2O2 (100µM) significantly
decreased cell viability and oleuropein pretreatment significantly prevented the cell
losses. Oleuropein regenerated total antioxidant capacity and glutathione levels decreased
by H2O2 exposure. In addition, nitric oxide and total oxidant capacity levels were also decreased
after administration of oleuropein in treated cells.
Conclusion:
Oleuropein was found to have potent antioxidative properties in human glioblastoma
cells. However, further studies and validations are needed in order to understand
the exact neuroprotective mechanism of oleuropein.
Carbonic anhydrase (CA) is a widely distributed enzyme and has a crucial role in the cells, tissues and organs of living organisms. It is found that CA-II is one of the most abundant CA isoenzymes in the gastrointestinal system. It plays an important role in the gastric acid secretion in stomach. In this study, we purified CA-II isoenzyme from sheep stomach with a 615.2 purification fold, 78% purification yield and 5562.02 specific activity. Moreover, the in vitro effects of some commonly used pesticides including chlorpyrifos, cypermethrin, dichlorvos, glyphosate isopropylamine and lambda cyhalomethrin on the enzyme activity were investigated. Of these compounds, glyphosate isopropylamine and dichlorvos showed an inhibition on CA-II esterase activity. They have IC50 values of 0.155 µM and 2.690 µM and Ki values of 0.329 µM and 3.654 µM, respectively. Both glyphosate isopropylamine and dichlorvos inhibited CA-II isoenzyme in a noncompetitive manner.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.