Acetic Acid → 4-Nitroaniline
Reaction equation
C1CN[C@@H](C(N[C@H](C(NC([C@@H]([NH:30][C:31]2[CH:36]=[CH:35][C:34]([N+:37]([O-:39])=[O:38])=[CH:33][CH:32]=2)CCCN=C(N)N)=O)=O)CC2C=CC=CC=2)=O)C1.C(O)(=O)C>C(O)C(N)(CO)CO.Cl>[N+:37]([C:34]1[CH:35]=[CH:36][C:31]([NH2:30])=[CH:32][CH:33]=1)([O-:39])=[O:38]Participants
Procedure
An inhibitor sample is dissolved in a 0.05 M Tris-hydrochloric acid buffer solution (pH=7.8) to make the total volume to 400 μl. To this solution, 50 μl of a 2 mM S-2302 solution is added and the mixture is incubated at a temperature of 37° C. for 5 minutes in a constant temperature bath. Then, 50 μl of a 0.12 unit/ml human plasma kallikrein is added and the mixture is incubated at a temperature of 37° C. for 5 minutes. Thereafter, 50 μl of 50% acetic acid is added to terminate the reaction. The absorbance of the p-nitroaniline formed in the reaction mixture is measured at 405 nm. Thus, the concentration I50 (μmol) of the inhibitor sample, at which the absorbance is one half (i.e., 1/2) of that obtained in the absence of the inhibitor sample, is determined.
Workup
- waitthe mixture is incubated at a temperature of 37° C. for 5 minutes
- customto terminate
- customthe reaction
Sources & evidence
- ORD record
- ord-9ef03242ae0d4da18105da159b70036d
- Source dataset
- uspto-grants-1989_10
- Patent number
- US04873253
Reaction notes
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