Pharmaceutical use of 1-(3-(6-chloropyridine)methyl)-3-phenyl-1H-pyrazol-5-carbohydrazide
A technology of carbohydrazide and chloropyridine, applied in the preparation of drugs for inhibiting vascular endothelial cell aging and apoptosis, field of 1-methyl)-3-phenyl-1H-pyrazole-5-carbohydrazide
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Embodiment 1
[0070] Example 1 Preparation of 1-(3-(6-chloropyridine)methyl)-3-phenyl-1H-pyrazole-5-carbohydrazide
[0071] 1) Add 0.690 gram of potassium carbonate (0.005 mol), 1.080 gram of 3-phenyl-1H-pyrazole-5-carboxylic acid ethyl ester (0.005 mol), 0.810 gram of 2-chloro-5- Chloromethylpyridine (0.005 mol) and acetonitrile (25 ml) were installed in a reflux condenser, and the upper part was connected to a drying tube. Heat to reflux for 4 hours, react until the raw material is completely consumed, and detect the end of the reaction by TLC. Concentrate under reduced pressure, remove the solvent, add ethyl acetate (30 ml) to dissolve the product, filter, the filtrate is concentrated, and use ethyl acetate-petroleum ether (V / V=1 / 2) as the eluent for silica gel column chromatography to separate the residue ( 100-200 mesh silica gel) to obtain ethyl 1-(3-(6-chloropyridine)methyl)-3-phenyl-1H-pyrazole-5-carboxylate;
[0072] 2) Add 1.2 Milliliter of 80% hydrazine hydrate was stirred and...
Embodiment 2
[0080] The cultured cells were divided into normal group cells, control group cells and treatment group cells.
[0081] Cells in the normal group were cultured with M199 medium containing serum and growth factors; cells in the control group were cultured without serum and growth factors; cells in the treatment group were treated with 1-(3-(6- Chloropyridine)methyl)-3-phenyl-1H-pyrazole-5-carbohydrazide treatment. After 24 hours of culture, the three groups of cells were fixed with 2% formaldehyde and 0.5% glutaraldehyde for 5 minutes, then washed once with the staining solution without β-galactose, and added the staining solution containing β-galactose at 37°C without carbon dioxide. Incubate under conditions for 18-24 hours. Observed after washing with PBS. Randomly select 10 fields of view for each of the above three groups of cells, count the total number of cells and the number of positive cells, and calculate the positive rate (see results under a microscope). Figure ...
Embodiment 3
[0085] The cultured cells were divided into normal group cells, control group cells and treatment group cells.
[0086] Cells in the normal group were cultured with M199 medium containing serum and growth factors; cells in the control group were cultured without serum and growth factors; cells in the treatment group were treated with 1-(3-(6- Chloropyridine)methyl)-3-phenyl-1H-pyrazole-5-carbohydrazide treatment. After the three groups of cells were cultured for 24 hours, the culture medium was discarded and fixed with 4% paraformaldehyde for 15 minutes. After washing with 0.1M PBS, normal serum blocking solution was added. Discard the blocking solution, add the primary antibody, discard the primary antibody and add the secondary antibody, keep at 37°C for 30 minutes, discard the secondary antibody, wash with 0.1M PBS and observe.
[0087] Fluorescent quantitative analysis was performed on the expression of cell membrane integrin β4 in the above three groups.
[0088] The r...
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