Back to Search
Start Over
Fragment-Based Discovery of Irreversible Covalent Inhibitors of Cysteine Proteases Using Chlorofluoroacetamide Library.
- Source :
-
Chemical & pharmaceutical bulletin [Chem Pharm Bull (Tokyo)] 2020; Vol. 68 (11), pp. 1074-1081. - Publication Year :
- 2020
-
Abstract
- Fragment-based approach combined with electrophilic reactive compounds is a powerful strategy to discover novel covalent ligands for protein target. However, the promiscuous reactivity often interferes with identification of the fragments possessing specific binding affinity to the targeted protein. In our study, we report the fragment-based covalent drug discovery using the chemically tuned weak reactivity of chlorofluoroacetamide (CFA). We constructed a small fragment library composed of 30 CFA-appended compounds and applied it to the covalent ligand screening for cysteine protease papain as a model protein target. Using the fluorescence enzymatic assay, we identified CFA-benzothiazole 30 as a papain inhibitor, which was found to irreversibly inactivate papain upon enzyme kinetic analysis. The formation of the covalent papain-30 adduct was confirmed using electrospray ionization mass spectrometry analysis. The activity-based protein profiling (ABPP) experiment using an alkynylated analog of 30 (i.e., 30-yne) revealed that 30-yne covalently labeled papain with high selectivity. These data demonstrate potential utility of the CFA-fragment library for de novo discovery of target selective covalent inhibitors.
- Subjects :
- Acetamides metabolism
Chromatography, High Pressure Liquid
Cysteine Proteases metabolism
Cysteine Proteinase Inhibitors metabolism
Drug Discovery
Fluorescent Dyes chemistry
Kinetics
Ligands
Papain antagonists & inhibitors
Papain metabolism
Spectrometry, Mass, Electrospray Ionization
Acetamides chemistry
Cysteine Proteases chemistry
Cysteine Proteinase Inhibitors chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1347-5223
- Volume :
- 68
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Chemical & pharmaceutical bulletin
- Publication Type :
- Academic Journal
- Accession number :
- 33132374
- Full Text :
- https://doi.org/10.1248/cpb.c20-00547