ABSTRACT
Several P4 domain derivatives of the general d-phenylglycinamide-based scaffold (2) were synthesized and evaluated for their ability to bind to the serine protease factor Xa. Some of the more potent compounds were evaluated for their anticoagulant effects in vitro. A select subset containing various P1 indole constructs was further evaluated for their pharmacokinetic properties after oral administration to rats.
Subject(s)
Antithrombin III/chemical synthesis , Antithrombin III/pharmacology , Glycine/analogs & derivatives , Anticoagulants/chemical synthesis , Anticoagulants/chemistry , Anticoagulants/pharmacology , Antithrombin III/chemistry , Crystallography, X-Ray , Factor Xa/chemistry , Factor Xa/metabolism , Glycine/chemical synthesis , Glycine/chemistry , Glycine/pharmacology , Humans , Models, Molecular , Molecular Structure , Protein Binding , Structure-Activity RelationshipABSTRACT
Analogs to a series of D-phenylglycinamide-derived factor Xa inhibitors were discovered. It was found that the S4 amide linkage can be replaced with an ether linkage to reduce the peptide character of the molecules and that this substitution leads to an increase in binding affinity that is not predicted based on modeling. Inhibitors which incorporate ether, amino, or alkyl S4 linkage motifs exhibit similar levels of binding affinity and also demonstrate potent in vitro functional activity, however, binding affinity in this series is strongly dependent on the nature of the S1 binding element.
Subject(s)
Anticoagulants/pharmacology , Factor Xa Inhibitors , Glycine/analogs & derivatives , Serine Proteinase Inhibitors/pharmacology , Anticoagulants/chemistry , Crystallography, X-Ray , Ethanolamines , Glycine/chemistry , Models, Molecular , Peptides/chemistry , Serine Proteinase Inhibitors/chemistryABSTRACT
SAR about the B-ring of a series of N(2)-aroyl anthranilamide factor Xa (fXa) inhibitors is described. B-ring o-aminoalkylether and B-ring p-amine probes of the S1' and S4 sites, respectively, afforded picomolar fXa inhibitors that performed well in in vitro anticoagulation assays.
Subject(s)
Antithrombin III/chemical synthesis , Factor Xa Inhibitors , Fluorescent Dyes/pharmacology , ortho-Aminobenzoates/pharmacology , Anticoagulants/chemistry , Antithrombin III/chemistry , Binding Sites , Chemistry, Pharmaceutical/methods , Dose-Response Relationship, Drug , Drug Design , Drug Evaluation, Preclinical , Fluorescent Dyes/chemical synthesis , Humans , Kinetics , Models, Chemical , Molecular Conformation , Structure-Activity Relationship , ortho-Aminobenzoates/chemical synthesisABSTRACT
The authors show by illustration that procedures used to validate the reliability of single-concentration high-throughput screens such as the signal window and Z' factor do not ensure sufficient reliability in potency estimates from concentration response assays. They develop the minimum significant ratio as a statistical parameter to characterize the fold change between 2 compounds run in the same experiment that can be considered a real difference and use this parameter to characterize the reliability of the assay. They adapt methods described by Bland and Altman to develop a simple set of 2 experiments to estimate the minimum significant ratio and show that this protocol can identify assays that lack reproducibility. The methods are then extended to validate the equivalency of the same assay run by multiple laboratories.
Subject(s)
Models, Statistical , Reproducibility of Results , Factor Xa/metabolism , Factor Xa InhibitorsABSTRACT
Several non-amidino S1 derivatives of the 1,2-diaminobenzene-based scaffold (4) were synthesized and evaluated for their ability to bind to the active site and inhibit the human protease factor Xa. A subset of these compounds were also evaluated for their anticoagulant effects in human plasma as measured by prothrombin time (PT).
Subject(s)
Anticoagulants/chemical synthesis , Benzene Derivatives/chemical synthesis , Factor Xa Inhibitors , Anticoagulants/pharmacology , Benzene Derivatives/pharmacology , Binding Sites , Blood Coagulation/drug effects , Factor Xa/metabolism , Humans , Models, Molecular , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Protein Binding , Prothrombin Time , Structure-Activity RelationshipABSTRACT
A novel isonitrile derivative was synthesized and used in an Ugi four component coupling reaction to explore aryl group substitution effects on inhibition of the coagulation cascade serine protease factor Xa.
