Urinary kallikrein assay: specific substrates and assay method
US4327178A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Mar 31, 1981 |
| Grant date | Apr 27, 1982 |
| Priority date | — |
| Expiry date | Mar 31, 2001 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S530/802
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Substrates and method for the assay of urinary kallikrein are provided. The substrates have the general formula ##STR1## wherein R is H, acetyl, benzoyl, cyclopentylcarbonyl, succinyl, R.sub.1 --Ser.sub.1 or R.sub.1 --Phe--Ser where R.sub.1 is H, acetyl, benzoyl, cyclopentylcarbonyl or succinyl, PA1 X is H, tritium, 3-iodo or 4-iodo, and PA1 n is 0 or 1. Radioactive label may be incorporated in the anilide or benzylamide moiety. Hydrolysis catalyzed by urinary kallikrein yields labeled aniline or benzylamine as product. The assay method includes mixing the enzyme with substrate in a buffered solution, pH 7.5-10.5, the substrate being present preferably at a concentration substantially below K.sub.m. After incubating to allow the reaction to proceed, the reaction is terminated and the radioactive hydrolysis product is separated and counted. Also disclosed are compounds of the type ##STR2## wherein R, R.sub.1 and n are as defined above. X is 3--OH or 4--OH. Y.sub.1 is 3-iodo if X is 4--OH or 6-iodo if X is 3--OH. Y.sub.2 may be H or 5-iodo if Y.sub.1 is 3-iodo, or 4-iodo if Y.sub.1 is 6-iodo.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.