RESUMO
Protein-templated reactions enable the target-guided formation of protein ligands from reactive fragments, ideally with no background reaction. Herein, we investigate the templated formation of amides. A nucleophilic fragment that binds to the coagulation factorâ Xa was incubated with the protein and thirteen differentially activated dipeptides. The protein induced a non-catalytic templated reaction for the phenyl and trifluoroethyl esters; the latter was shown to be a completely background-free reaction. Starting from two fragments with millimolar affinity, a 29â nm superadditive inhibitor of factorâ Xa was obtained. The fragment ligation reaction was detected with high sensitivity by an enzyme activity assay and by mass spectrometry. The reaction progress and autoinhibition of the templated reaction by the formed ligation product were determined, and the structure of the protein-inhibitor complex was elucidated.
Assuntos
Amidas/química , Dipeptídeos/química , Inibidores do Fator Xa/química , Fator Xa/química , Amidas/síntese química , Amidas/farmacologia , Benzamidinas/síntese química , Benzamidinas/química , Benzamidinas/farmacologia , Técnicas de Química Sintética/métodos , Dipeptídeos/síntese química , Dipeptídeos/farmacologia , Descoberta de Drogas , Esterificação , Fator Xa/metabolismo , Inibidores do Fator Xa/síntese química , Inibidores do Fator Xa/farmacologia , Humanos , Ligantes , Simulação de Acoplamento MolecularRESUMO
Protein-templated fragment ligation is a novel concept to support drug discovery and can help to improve the efficacy of protein ligands. Protein-templated fragment ligations are chemical reactions between small molecules ("fragments") utilizing a protein's surface as a reaction vessel to catalyze the formation of a protein ligand with increased binding affinity. The approach exploits the molecular recognition of reactive small-molecule fragments by proteins both for ligand assembly and for the identification of bioactive fragment combinations. In this way, chemical synthesis and bioassay are integrated in one single step. This Review discusses the biophysical basis of reversible and irreversible fragment ligations and gives an overview of the available methods to detect protein-templated ligation products. The chemical scope and recent applications as well as future potential of the concept in drug discovery are reviewed.
