A serendipitously identified novel small molecule procoagulant compound giving rise to a high-throughput screening assay based on human plasma

作者：Gustafsson David^*; Nielsen Sofi; McPheat Jane; Wagberg Fredrik; Kaspersson Karin; Rohman Mattias; Ulander Johan; Fex Tomas; Fjellstroem Ola; Deinum Johanna

来源：Thrombosis Research, 2013, 132(2): 248-255.

DOI：10.1016/j.thromres.2013.05.023

• Haemostasis
• Haemophilia
• Heparin
• Thrombin

Introduction: 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.
Material and 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 mu M (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.
Conclusion: These novel small molecule procoagulant compounds may serve as templates for discovery of oral procoagulant drugs.

• 出版日期2013-8