OSELTAMIVIR POTTASIUM CARBOXYLATE BDBM50484773 Oseltamivir Potassium Carboxylate
Guanidino-Oseltamivir Carboxylicacid BDBM50028503 CHEMBL81717
BDBM5025 Oseltamivir ethyl (3R,4R,5S)-5-amino-4-acetamido-3-(pentan-3-yloxy)cyclohex-1-ene-1-carboxylate US10919856, POSITIVE CONTROL
GS4071 Oseltamivir carboxylate BDBM4994 CHEMBL674 (3R,4R,5S)-5-amino-4-acetamido-3-(pentan-3-yloxy)cyclohex-1-ene-1-carboxylic acid
US9527857, M BDBM253921 US9464092, M
BDBM204122 US9539260, M US9763952, Example M
BDBM256611 US9790174, Example 85(m) US10435361, Example 85(m) US9481648, 85(m)
US11267820, Compound m US10689395, Compound m BDBM448086
1,3-xylene 1,3-Dimethylbenzol m-methyltoluene BDBM50008556 m-xylene CHEMBL286727 m-Xylol 3-xylene meta-xylene m-dimethylbenzene
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- Neuraminidase Inhibition Assay The 50% inhibitory concentration (IC50) determination was carried out by using a 96-well microtitre plate as an array of reaction vessels. In the first well,a 0.1-mM solution of oseltamivir carboxylate was prepared by mixing 90 μl of 33 mM MES pH 6.5 containing 4 mM CaCl2 and 10 μl of a 1-mM oseltamivir carboxylate stock solution. Starting from the solution contained in the first well, an in-plate 10-fold serial dilution was performed using 33 mM MES pH 6.5 containing 4 mM CaCl2 as dilution buffer obtaining a 7 points dilution curve, the last well in the vertical lane was used to obtain a negative control (no oseltamivir was added). At the end of the dilution process, each well contained 90 μl of solution. To each well, 25 μl of the enzyme stock solution were mixed with the oseltamivir solution and incubated for 2 h at 37°C. After the incubation time, 25 μl of 20 μM MUNANA were added. In the reaction mixtures, the final concentration of oseltamivir carboxylate spanned in the ranges 0.064 nM-64.2 μM. After incubating the plate for 5 h at 37°C, 50 μl of stop solution (0.1 M Glycine, pH 10.7 containing 25% ethanol) were added. The fluorescence of the solutions contained in each well was read with the Victor3 multi-label counter. For the determination of the of oseltamivir carboxylate against neuraminidase N3, the same procedure described above was used, except for the concentrationof the oseltamivir carboxylate stock solution which was 10 μM. In the reaction mixtures, the final concentration of oseltamivir carboxylate spanned in the ranges 0.64 pM-642 nM.
- ChEMBL_1295475 (CHEMBL3131473) Inhibition of oseltamivir-resistant Influenza A virus H1N1 B/55/08 neuraminidase by chemiluminescence based assay
- ChEMBL_819005 (CHEMBL2034237) Inhibition of oseltamivir-resistant H1N1 swine influenza virus neuraminidase H274Y mutant using 4-MU-NANA as substrate by fluorescence assay
- ChEMBL_698045 (CHEMBL1645796) Inhibition of oseltamivir-resistant H1N1 swine influenza virus neuraminidase H274Y mutant activity expressed in HEK293T cells after 2 hrs by spectrofluorometry
- ChEMBL_675041 (CHEMBL1272775) Inhibition of oseltamivir-resistant Influenza A virus (A/California/08/2009(H1N1)) neuraminidase H274Y mutant expressed in human 293T cells after 30 mins by spectrofluorimetric analysis
- Neuraminidase Activity Assay Practically in a microtitre plate, 90 μl of 33 mM MES pH 6.5 containing 4 mM CaCl2 were dispensed in each well, except for the first well of each lane. In the first well of each vertical lane, 18 μl of 0.01 mM oseltamivir were mixed with 162 μl of 33 mM MES pH 6.5 containing 4 mM CaCl2. The solution was thoroughly mixed to obtain a 1-μM oseltamivir carboxylate solution. For each assay, a 90-μl aliquot of this solution was withdrawn from the first well and added to the second well in the same vertical lane obtaining a 2-fold diluted solution of oseltamivir. The 2-fold dilution process was continued up to the seventh well, while the eighth well used as blank (no inhibitor was added). At the end of the dilution process, each well contained 90 μl of solution. To each well, 25 μl of enzyme stock solution were added. After an incubation time of 2 h, 25 μl of MUNANA were added to each well, the final concentrations of oseltamivir carboxylate spanned the range 10-643 nM for N1 assay. After 30 min of incubation, 50 μl of stop solution were added to the first vertical lane, and the fluorescence was recorded. After an additional 30 min (60 min total), the stop solution was added to the second vertical lane and the fluorescence recorded. After additional 30 min (90 min total), the stop solution was added to the third vertical lane and the fluorescence recorded and so on. The experiment was performed in duplicate. For the determination of oseltamivir carboxylate Ki against N3, the same procedure described for N1 was used, except for the concentration of oseltamivir carboxylate stock solution which was 0.1 μM.