In vitro characterization of CT‐001—a short‐acting factor VIIa with enhanced prohemostatic activity

Sim, Derek; Mallari, Cornell; Teare, John; Feldman, Richard I.; Bauzon, Maxine; Hermiston, Terry

doi:10.1002/rth2.12530

Cited by 4 publications

(14 citation statements)

References 59 publications

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“…23 Therefore, along with the current treatment regimen, it is desirable to have a therapeutic agent that could provide patients with additional prohemostatic activity for treating blood loss while having low thrombogenicity risk by being quickly cleared. CT-001, through two newly engineered amino acids in the γ-carboxyglutamic acid domain to provide enhanced clotting activity and 4 N-glycosylation sites expressing desialylated glycans for rapid removal by hepatocytes, 12 may represent such a needed treatment. In this study, N-glycan profile analyses of CT-001 indicated that all four N-glycosylation sites expressed glycan structures.…”

Section: Discussionmentioning

confidence: 99%

“…The blood flow in liver was reduced to 30% of control. CT-001 is known to be cleared rapidly by hepatocytes 12 . While this reduced blood flow is expected to be sufficient for maintaining the hepatocyte-mediated rapid clearance of CT-001, the PK of CT-001 under traumatic shock remains to be experimentally defined.…”

Section: Discussionmentioning

confidence: 99%

“…N -acetylneuraminic acid (NANA), N -glycolylneuraminic acid (NGNA), 4,5-methylenedioxy-1,2-phenylenediamine dihydrochloride (DMB), and 4% sodium citrate were purchased from Sigma (St. Louis, MO); human tissue factor knockin (TFKI) mice, from Jackson Laboratory (Sacramento, CA); Sprague-Dawley rats and cynomolgous monkeys, from Charles River (Reno, NV); normal human plasma, from George King Bio-Medical, Inc. (Overland Park, KS); anti-FVIIa, biotinylated anti-FVIIa, and HRP-streptavidin, 1-Step Ultra TMB, from ThermoFisher (Waltham, MA); APC and thrombomodulin, from Prolytix (Essex Junction, VT); STA-PTT Automate 5, STA Neoplastin CI Plus 5, MP-Reagent, PPP-Reagent LOW, Thrombin Calibrator, and FluCa kit, from Stago (Parsipanny, NJ); ketamine and dexmedetomidine, from Zoetis Animal Health (Parsippany, NJ); WT FVIIa (NovoSeven, eptacog alfa activated), from Novo Nordisk (Bagsvaerd, Denmark); and TXA, from Pfizer (New York, NY). CT-001 was generated as described previously 12 …”

Section: Methodsmentioning

confidence: 99%

“…CT-001 is known to be cleared rapidly by hepatocytes. 12 While this reduced blood flow is expected to be sufficient for maintaining the hepatocyte-mediated rapid clearance of CT-001, the PK of CT-001 under traumatic shock remains to be experimentally defined. Large species, such as pigs, would be important TIC models for future studies.…”

Section: Acknowledgmentmentioning

confidence: 99%

“…This predicament could potentially be addressed by using a FVIIa that could be cleared before the development of a prothrombotic state. CT-001 is a novel FVIIa molecule with P10Q/K32E amino acid substitutions in the γ-carboxyglutamic acid domain for enhanced procoagulant activity and 4 N-glycosylation sites (T106N/N145/V253N/N322) expressing desialylated glycans for rapid hepatocyte clearance 12 . The efficacy and low thrombogenicity of CT-001 in mouse models have been previously reported 13 .…”

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confidence: 99%

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Rapid clearing CT-001 restored hemostasis in mice with coagulopathy induced by activated protein C

Sim¹,

Mallari²,

Bauzon³

et al. 2023

J Trauma Acute Care Surg

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BACKGROUND: Activated protein C (APC) is one of the mechanisms contributing to coagulopathy, which is associated with high mortality. The counteraction of the APC pathway could help ameliorate bleeding. However, patients also transform frequently from a hemorrhagic state to a prothrombotic state at a later time. Therefore, a prohemostatic therapeutic intervention should take this thrombotic risk into consideration. OBJECTIVES:CT-001 is a novel factor VIIa (FVIIa) with enhanced activity and desialylated N-glycans for rapid clearance. We assessed CT-001 clearance in multiple species and its ability to reverse APC-mediated coagulopathic blood loss. METHODS:The N-glycans on CT-001 were characterized by liquid chromatography-mass spectrometry. Three species were used to evaluate the pharmacokinetics of the molecule. The potency and efficacy of CT-001 under APC pathway-induced coagulopathic conditions were assessed by coagulation assays and bleeding models. RESULTS:The N-glycosylation sites of CT-001 had high occupancy of desialylated N-glycans. CT-001 exhibited 5 to 16 times higher plasma clearance in human tissue factor knockin mice, rats, and cynomolgus monkeys than wildtype FVIIa. CT-001 corrected the activated partial thromboplastin time and thrombin generation of coagulopathic plasma to normal in in vitro studies. In an APC-mediated saphenous vein bleeding model, 3 mg/kg of CT-001 reduced bleeding time in comparison with wildtype FVIIa. The correction of bleeding by CT-001 was also observed in a coagulopathic tail amputation severe hemorrhage mouse model. The efficacy of CT-001 is independent of the presence of tranexamic acid, and the combination of CT-001 and tranexamic acid does not lead to increased thrombogenicity. CONCLUSION: CT-001 corrected APC pathway-mediated coagulopathic conditions in preclinical studies and could be a potentially safe and effective procoagulant agent for addressing APC-mediated bleeding.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Acknowledgmentmentioning

confidence: 99%

mentioning

confidence: 99%

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Rapid clearing CT-001 restored hemostasis in mice with coagulopathy induced by activated protein C

Sim¹,

Mallari²,

Bauzon³

et al. 2023

J Trauma Acute Care Surg

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Site-directed mutagenesis of tissue factor pathway inhibitor–binding exosite D60A on factor VII results in a new factor VII variant with lower coagulant activity

Seanoon,

Kitiyanant,

Payongsri

et al. 2024

Research and Practice in Thrombosis and Haemostasis

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CT-001, a novel fast-clearing factor VIIa, enhanced the hemostatic activity in postpartum samples

Sim,

Mallari,

Hermiston

et al. 2024

Blood Advances

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The hemostatic system is upregulated to protect pregnant mothers from hemorrhage during childbirth. Studies of the details just prior to and after delivery, however, are lacking. Recombinant factor VIIa (rFVIIa) has recently been granted approval by the European Medicines Agency for the treatment of postpartum hemorrhage (PPH). A next generation molecule, CT-001, is being developed as a potentially safer and more efficacious rFVIIa-based therapy. We sought to evaluate the peripartum hemostatic status of pregnant women and assess the ex-vivo hemostatic activity of rFVIIa and CT-001in peripartum blood samples. Pregnant women from 2 study sites were enrolled in this prospective observational study. Baseline blood samples were collected up to 3 days before delivery. Post-delivery samples were collected 45 (+/-15) mins after delivery. Between the 2 timepoints, soluble fibrin monomer and D-dimer increased while tissue factor, factor VIII, factor V, and fibrinogen decreased. Interestingly, the post-delivery lag time and time-to-peak in the thrombin generation assay were shortened, and the peak thrombin generation capacity was maintained despite the reduced levels of coagulation proteins post-delivery. Furthermore, both rFVIIa and CT-001 were effective in enhancing clotting activity of post-delivery samples in APTT, PT, TGA, and viscoelastic hemostatic assays, with CT-001 demonstrating greater activity. In conclusion, despite apparent ongoing consumption of coagulation factors at the time of delivery, thrombin output was maintained. Both rFVIIa and CT-001 enhanced the upregulated hemostatic activity in post-delivery samples and, consistent with previous studies comparing CT-001 and rFVIIa in vitro and in in vivo, CT-001 demonstrated greater activity than rFVIIa.

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In vitro characterization of CT‐001—a short‐acting factor VIIa with enhanced prohemostatic activity

Cited by 4 publications

References 59 publications

Rapid clearing CT-001 restored hemostasis in mice with coagulopathy induced by activated protein C

Rapid clearing CT-001 restored hemostasis in mice with coagulopathy induced by activated protein C

Site-directed mutagenesis of tissue factor pathway inhibitor–binding exosite D60A on factor VII results in a new factor VII variant with lower coagulant activity

CT-001, a novel fast-clearing factor VIIa, enhanced the hemostatic activity in postpartum samples

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