Application of 3-[4-(sulfonyl)benzene] urea compound in preparation of medicine for tumour
A compound and urea technology, applied in the field of 3-[4-phenyl]urea compounds, can solve the problem of limitation and achieve the effect of proliferation inhibition
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[0027] Example 1: Analysis of 1-(3-chloro-4-fluorophenyl)-3-[4-(N-methylsulfonamido)phenyl]urea Enzyme inhibition activity assay of ROCK1. Enzyme inhibitory activity assay of ROCK1
[0028] Experimental principle: Invitrogen’s Z′-LYT technology was used to measure the ROCK1 inhibitory activity of the compound. This technology is based on the principle of fluorescence resonance energy transfer (FRET) and is based on the difference in sensitivity of phosphorylated and non-phosphorylated peptides to proteolytic cleavage. .
[0029] experimental method:
[0030] (1) Preparation of reagents. Kinase buffer: dilute 2ml of 5X kinase buffer to 10ml with water; compound: dilute the compound to be tested with water to a concentration gradient; ROCK kinase / substrate mixed solution: prepare 2250 μL of ROCK / substrate mixed solution; The concentration of enzyme is 10ng / ml, the concentration of substrate is 4μM, and the solvent is kinase buffer; phosphorylated peptide solution: add 2μL o...
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