Comparison of the sensitivity of commercial aPTT tests in measurement of the pharmacodynamic response of milvexian, a novel FXIa inhibitor

Abstract Introduction The activated partial thromboplastin time (aPTT) assay is a test routinely used to evaluate abnormalities or deficiencies in coagulation factors of the intrinsic and common pathways. The composition of the surface activator and phospholipids in the aPTT reagent is known to influence the response of heparin and direct FXa or thrombin inhibitors. Milvexian (formerly referred to as BMS-986177/JNJ-70033093) is an investigational small-molecule Factor XIa (FXIa) inhibitor being studied for the prevention and treatment of major thrombotic conditions. Clinical pharmacology evaluation of milvexian includes aPTT as the primary pharmacodynamic assay, thus underscoring the need for a sensitive reagent to evaluate its anticoagulant activity. Purpose This study evaluated the sensitivity of six commercially available aPTT reagents in plasma from individual donors spiked with milvexian, at concentrations spanning the anticipated clinically relevant exposures. Methods Platelet-poor plasma (PPP) prepared from citrated whole blood collected from consenting, healthy adult volunteers (n=12) was spiked with vehicle (DMSO, 0.5% v/v) or 0.1–10 μM milvexian. The aPTT was measured using six commercially available aPTT reagent kits which included silica; kaolin, or ellagic acid as the contact activator, in combination with natural or synthetic phospholipids. The coagulation automated analysers used for testing were matched to the kit's manufacturer recommendations. All reagents and respective normal or abnormal controls were prepared as instructed by the manufacturer. Results Milvexian exhibited dose-dependent prolongation of aPTT with all reagents tested. Assays performed with aPTT reagents containing kaolin or ellagic acid demonstrated the highest sensitivity, as measured by the concentration that achieved a 2-fold aPTT prolongation (EC2x), and showed the widest dynamic range of response. Coefficient of variability of aPTT measurements in plasma from 12 individual donors was between 5.6–7.9% for Dade® Actin® FS, and 7.1–8% for STA®-C.K. Prest® 5. Conclusions Prolongation of aPTT in PPP spiked with milvexian exhibited a dose-dependent relationship, with statistically significant differences observed among reagents at milvexian concentrations above 0.3 μM. The highest sensitivity measured by changes in the ratio to baseline was obtained with aPTT reagents containing an activator/procoagulant phospholipid combination of ellagic acid + purified soy phosphatides