| Assay Method Information | |
| | Inhibitory Assay |
| Description: | The inhibitory activity of the compounds of the present invention for recombinant human chymase was measured by the method of Pasztor et al. (Pasztor et al., Acta. Biol. Hung. 42: 285-95, 1991). That is, recombinant human chymase was diluted to an appropriate concentration by a 50 mM tris-hydrochloride buffer (pH 7.5), 1M sodium chloride solution, and 0.01% (v/v) Triton X-100 to obtain an enzyme solution. A 10 mM dimethyl sulfoxide (hereinafter referred to as DMSO) solution of Suc-Ala-Ala-Pro-Phe-MCA (Peptide Institute) was diluted 20-fold at the time of use by 50 mM tris-hydrochloride buffer (pH 7.5), 1M sodium chloride solution and 0.01% (v/v) Triton X-100 to obtain the substrate solution. 75 ul of the enzyme solution was mixed with 5 ul of the test compound in a DMSO solution, and incubated for 10 minutes. Then 20 ul of the substrate solution was added to the mixture and reacted for a further 10 minutes at room temperature. |
| Affinity data for this assay | |
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