title: β-D-glucosyl conjugates of highly potent inhibitors of blood coagulation factor Xa bearing 2-chorothiophene as a P1 motif.
abstract: We synthesized a novel O-glucoside of the recently reported potent factor Xa (fXa) inhibitor 1, which bears a 5-chlorothien-2-yl moiety and 1-isopropylpiperidine as fragments that bind the S1 and S4 enzyme pockets, respectively. A β-D-glucosyl unit was conjugated through an ether-linked C3-alkyl spacer to the central phenyl ring of 1. The synthesized β-D-glucose-based compound 16 achieved picomolar inhibitory potency against human fXa (K(i)=60 pM) and high selectivity over thrombin and other serine proteases. In addition to the chlorothienyl S1 binder, a large gain in ΔG resulted from the addition of protonated 1-isopropylpiperidine (ΔΔG=29.7-30.5 kJ mol(-1)), which should bind to the aromatic S4 pocket through efficient cation-π and C-H···π interactions. Instead, the C3-alkyl-linked glucose fragment, which is likely directed toward the solvent outside the enzyme binding site, improves ΔG by an average of 2.9-3.8 kJ mol(-1) . Compound 16 showed sub-micromolar in vitro anticoagulant activity, as assessed by prothrombin time (PT) and activated thromboplastin time (aPTT) clotting assays in pooled human plasma (PT(2) and aPTT(2) equal to 0.135 and 0.389 μM, respectively). Although compound 16 was 1.4-fold less active than parent compound 1 in the ex vivo anticoagulant assay in mice, it showed a significant (1.6-fold) prolongation of PT relative to controls (P<0.05) 60 min after oral dosing (75 mg kg(-1)).
authors.affiliation: Dipartimento Farmaco Chimico, University of Bari A. Moro, Via E. Orabona 4, 70125 Bari, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Lopopolo
authors.forename: Modesto
authors.initials: M
authors.lastname: de Candia
authors.forename: Luigi
authors.initials: L
authors.lastname: Panza
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: Romano
authors.forename: Marcello Diego
authors.initials: MD
authors.lastname: Lograno
authors.forename: Francesco
authors.initials: F
authors.lastname: Campagna
authors.forename: Cosimo
authors.initials: C
authors.lastname: Altomare
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D001779:Blood Coagulation Factors / Q000037:antagonists & inhibitors / Q000378:metabolism; D001780:Blood Coagulation Tests; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D005947:Glucose / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D008297:Male; D051379:Mice; D013876:Thiophenes / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D000925:Anticoagulants; D001779:Blood Coagulation Factors; D004791:Enzyme Inhibitors; D065427:Factor Xa Inhibitors; D013876:Thiophenes; D015951:Factor Xa; D005947:Glucose