Query String: LEVAMISOLE
R-12564 Levamisole CHEMBL1770 LEVAMISOLE HYDROCHLORIDE BDBM50304534 Ergamisol
cid_68628 6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole BDBM50241179 6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole(Levamisole) TCMDC-125847 6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole (Levamisole) CHEMBL1454 LEVAMISOLE HYDROCHLORIDE levamisole;6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole (S)-6-phenyl-2,3,5,6-tetrahydroimidazo[2,1-b]thiazole tetramisole;6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole 6-Phenyl-2,3,5,6-tetrahydro-imidazo[2,1-b]thiazole(tetramisole) LEVAMISOLE
- Alkaline Phosphatase Assay Alkaline phosphatase assay was optimized and performed in the same way as previously reported method with slight modifications [Sergienko et al., Nat. Protoc., 5:1431-1439]. All compounds were analyzed at a final concentration of 200 μM (1% DMSO) by using assay buffer with composition of 8 M diethanolamine (DEA) (pH 9.8), 0.05 mM ZnCl2 and 2.5 mM MgCl2. The total reaction volume was kept 50 μL containing 10 μL of tested compound, 20 μL of b-TNAP (1:800 (0.8 units/mL) diluted enzyme in assay buffer) or c-IAP (1:800 (1 units/mL) diluted enzyme in assay buffer). After incubation for 3-5 min at 37 °C., the luminescence was measured as a pre-read using microplate reader (BioTek FLx800, Instruments, Inc. USA). Then, 20 μL of substrate (CDPstar®) (final concentration of 110 μM) was added to the reaction mixture to initiate the enzymatic reaction. The change in the luminescence was recorded after 15 min. at 37 °C. The activity of each compound was measured in comparison with control (without any inhibitor). Levamisole (2 mM per well) and L-phenylalanine (4 mM per well) were used as standards (positive control) against b-TNAP) and c-IAP), respectively.
- Alkaline Phosphatase Inhibition Assay The assay conditions were optimized with some changes in formerly reported spectrophotometric method. The composition of assay buffer of pH 9.8 was as: 8 M diethanolamine (DEA), 0.05 mM ZnCl2 and 2.5 mM MgCl2. All compounds were tested at the initial concentration of 0.2 mM. A total assay volume of 50 μL contained 10 μL of tested compound, followed by the addition of 20 μL of either b-TNAP (1:800 times diluted enzyme in assay buffer) or 20 μL of c-IAP (1:800 times diluted enzyme in same assay buffer). After incubating the reaction mixture for 3-5 minutes at 37 °C, pre-read was taken by measuring luminescence with the help of microplate reader (BioTek FLx800, Instruments, Inc. USA). After that, 20 μL of substrate (CDP-Star) at the concentration of (110 μM) was added to the reaction mixture. Then the reaction mixture was again incubated for 15-20 min at 37 °C and after read was taken to measure the change in the luminescence. Total activity control (without inhibitor or negative control) was used for comparison of the activity of each test compound. L-phenylalanine (at concentration of 4 mM per well) and Levamisole (at concentration of 2 mM per well) were used as reference standards against calf intestinal alkaline phosphatase (c-IAP) and bovine tissue-nonspecific alkaline phosphatase (b-TNAP) respectively.
- Alkaline Phosphatase Inhibition Assay Alkaline phosphatase (b-TNAP, c-IAP purchased from Calzyme Laboratories, Inc. USA) inhibition assay of different cyclic sulfonamides was performed using a chemiluminescent substrate i.e. CDP-Star® (disodium 2-chloro-5-(4-methoxyspiro[1,2-dioxetane-3,2'-(5-chlorotricyclo[3.3.1.13.7]decan])-4-yl]-1-phenyl phosphate). With a slight modification in reported spectrophotometric procedure, an optimized conditions of the bioassay were developed [Hessle et al., Proc. Natl. Acad. Sci., 99:9445-9449]. Buffer solution for this bioassay was composed of 8 M DEA (pH 9.8) that contain 2.5 mM MgCl2 and 0.05 mM ZnCl2. First of all, the compounds were tested at the final concentration of 200 μM(with final DMSO 1% (v/v)). In bioassay procedure, the total volume (50 μL solution) contained 10 μL of tested compound, followed by the addition of 20 μL of TNAP (1:800 times diluted (0.8 unit/mL) enzyme in assay buffer) or 20 μL of IAP (1:800 times diluted (1 unit/mL) enzyme in assay buffer). The mixture was allowed to incubate for 3-5 min at 37 °C and luminescence was measured using microplate reader (BioTek FLx800, Instruments,Inc. USA). The reaction was initiated by the addition of 20 μL of CDP-star® (final concentration of 110 μM) and the assay mixture was incubated again at 37 °C. The change in the luminescence was measured after for 15 min of incubation. The activity of each compound was compared with total activity control (without any inhibitor). Levamisole (2 mM per well) and L-phenylalanine (4 mM per well) were used as a standard inhibitors for tissuenonspecific alkaline phosphatase (TNAP) and calf intestinal alkaline phosphatase (IAP), respectively.