title: A serendipitously identified novel small molecule procoagulant compound giving rise to a high-throughput screening assay based on human plasma.
abstract: BACKGROUND
  Oral treatment is lacking for haemophilia, the rare bleeding disorders, and some severe forms of von Willebrand's disease. We have serendipitously identified a small molecule procoagulant compound (AZ10047130). This publication describes some characteristics of AZ10047130 and a systematic search for novel hits using a, human plasma-based, high-throughput screening (HTS) assay.
  
  
  METHODS
  Coagulation, thrombin generation, chromogenic assays and surface plasmon resonance (SPR) experiments were used to characterise AZ10047130. A 1536-well formatted human plasma coagulation assay for HTS was developed.
  
  
  RESULTS
  In the plasma clot assay (re-calcified plasma with low tissue factor) AZ10047130 shortened time to coagulation with an EC50 value of 3.9 μM (assay concentration). AZ10047130 was similarly effective in immunodepleted human and haemophilia A plasmas. SPR and chromogenic substrate experiments indicated that AZ10047130 binds to the heparin binding site of several coagulation factors. The HTS screened in excess of one million compounds. It generated some hits belonging to the same pharmacophore as AZ10047130 but also some entirely novel hits.
  
  
  CONCLUSIONS
  These novel small molecule procoagulant compounds may serve as templates for discovery of oral procoagulant drugs.
authors.affiliation: Department of Bioscience, AstraZeneca, R&D Mölndal, Sweden. Electronic address: david.gustafsson@astrazeneca.com.
authors.forename: David
authors.initials: D
authors.lastname: Gustafsson
authors.forename: Sofi
authors.initials: S
authors.lastname: Nielsen
authors.forename: Jane
authors.initials: J
authors.lastname: McPheat
authors.forename: Fredrik
authors.initials: F
authors.lastname: Wågberg
authors.forename: Karin
authors.initials: K
authors.lastname: Kaspersson
authors.forename: Mattias
authors.initials: M
authors.lastname: Rohman
authors.forename: Johan
authors.initials: J
authors.lastname: Ulander
authors.forename: Tomas
authors.initials: T
authors.lastname: Fex
authors.forename: Ola
authors.initials: O
authors.lastname: Fjellström
authors.forename: Johanna
authors.initials: J
authors.lastname: Deinum
mesh_terms: D001572:Benzofurans / Q000737:chemistry / Q000494:pharmacology*; D001774:Blood Chemical Analysis / Q000379:methods*; D001777:Blood Coagulation / Q000187:drug effects*; D001779:Blood Coagulation Factors / Q000737:chemistry / Q000494:pharmacology*; D006467:Hemophilia A / Q000097:blood / Q000188:drug therapy; D006487:Hemostasis / Q000187:drug effects; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology*; D013917:Thrombin / Q000096:biosynthesis
chemical_list: C000588883:AZ10047130; D001572:Benzofurans; D001779:Blood Coagulation Factors; D013449:Sulfonamides; D013917:Thrombin
keywords: AT; AZ; AstraZeneca; CAT; ETP; HTS; Haemophilia; Haemostasis; Heparin; LT; PPP; PRP; RU; SPR; TF; TFPI; Thrombin; antithrombin; calibrated automated thrombogram; endogenous thrombin potential; high-throughput screening; lag time; platelet poor plasma; platelet rich plasma; resonance units; surface plasmon resonance; time to peak; tissue factor; tissue factor pathway inhibitor; ttPeak; vWD; von Willebrand Disease