In Vitro Characterization of CT-001-a Short-acting Factor VIIa with Enhanced Prohemostatic Activity

Overview

Journal Res Pract Thromb Haemost

Publisher Elsevier

Date 2021 Jul 15

PMID 34263099

Citations 4

Authors

Derek S Sim

Cornell R Mallari

John M Teare

Richard I Feldman

Maxine Bauzon

Terry W Hermiston

Affiliations

Soon will be listed here.

Abstract

Background: Traumatic injury and the associated acute bleeding are leading causes of death in people aged 1 to 44 years. Acute bleeding in pathological and surgical settings also represents a significant burden to the society. Yet there are no approved hemostatic drugs currently available. While clinically proven as an effective pro-coagulant, activated factor VII (FVIIa) use in acute bleeding has been hampered by unwanted thromboembolic events. Enhancing the ability of FVIIa to quickly stop a bleed and clear rapidly from circulation may yield an ideal molecule suitable for use in patients with acute bleeding.

Objectives: To address this need and the current liability of FVIIa, we produced a novel FVIIa molecule (CT-001) with enhanced potency and shortened plasma residence time by cell line engineering and FVIIa protein engineering for superior efficacy for acute bleeding and safety.

Methods: To address safety, CT-001, a FVIIa protein with 4 desialylated N-glycans was generated to promote active recognition and clearance via the asialoglycoprotein receptor. To enhance potency, the gamma-carboxylated domain was modified with P10Q and K32E, which enhanced membrane binding.

Results: Together, these changes significantly enhanced potency and clearance while retaining the ability to interact with the key hemostatic checkpoint proteins antithrombin and tissue factor pathway inhibitor.

Conclusions: These results demonstrate that a FVIIa molecule engineered to combine supra-physiological activity and shorter duration of action has the potential to overcome the current limitations of recombinant FVIIa to be a safe and effective approach to the treatment of acute bleeding.

Citing Articles

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 K, Kitiyanant V, Payongsri P, Sirachainan N, Angchaisuksiri P, Chuansumrit A Res Pract Thromb Haemost. 2024; 8(1):102309.

PMID: 38318153 PMC: 10840347. DOI: 10.1016/j.rpth.2023.102309.

The Role of the Complement System in the Pathogenesis of Infectious Forms of Hemolytic Uremic Syndrome.

Avdonin P, Blinova M, Generalova G, Emirova K, Avdonin P Biomolecules. 2024; 14(1).

PMID: 38254639 PMC: 10813406. DOI: 10.3390/biom14010039.

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

Sim D, Mallari C, Hermiston T, Bae D, Lee S, Allen T Blood Adv. 2023; 8(2):287-295.

PMID: 38039512 PMC: 10824690. DOI: 10.1182/bloodadvances.2023011398.

In vitro characterization of CT-001-a short-acting factor VIIa with enhanced prohemostatic activity.

Sim D, Mallari C, Teare J, Feldman R, Bauzon M, Hermiston T Res Pract Thromb Haemost. 2021; 5(5):e12530.

PMID: 34263099 PMC: 8265787. DOI: 10.1002/rth2.12530.

References

Persson E, Olsen O, Bjorn S, Ezban M . Vatreptacog alfa from conception to clinical proof of concept. Semin Thromb Hemost. 2012; 38(3):274-81. DOI: 10.1055/s-0032-1302442. View

Mayer S, Brun N, Begtrup K, Broderick J, Davis S, Diringer M . Recombinant activated factor VII for acute intracerebral hemorrhage. N Engl J Med. 2005; 352(8):777-85. DOI: 10.1056/NEJMoa042991. View

Mahlangu J, Coetzee M, Laffan M, Windyga J, Yee T, Schroeder J . Phase I, randomized, double-blind, placebo-controlled, single-dose escalation study of the recombinant factor VIIa variant BAY 86-6150 in hemophilia. J Thromb Haemost. 2012; 10(5):773-80. DOI: 10.1111/j.1538-7836.2012.04667.x. View

Lentz S, Ehrenforth S, Abdul Karim F, Matsushita T, Weldingh K, Windyga J . Recombinant factor VIIa analog in the management of hemophilia with inhibitors: results from a multicenter, randomized, controlled trial of vatreptacog alfa. J Thromb Haemost. 2014; 12(8):1244-53. PMC: 4238784. DOI: 10.1111/jth.12634. View

Lodge J, Jonas S, Jones R, Olausson M, Mir-Pallardo J, Soefelt S . Efficacy and safety of repeated perioperative doses of recombinant factor VIIa in liver transplantation. Liver Transpl. 2005; 11(8):973-9. DOI: 10.1002/lt.20470. View

Nelsestuen G, Stone M, Martinez M, Harvey S, Foster D, Kisiel W . Elevated function of blood clotting factor VIIa mutants that have enhanced affinity for membranes. Behavior in a diffusion-limited reaction. J Biol Chem. 2001; 276(43):39825-31. DOI: 10.1074/jbc.M104896200. View

Goodnough L, Levy J . The Judicious Use of Recombinant Factor VIIa. Semin Thromb Hemost. 2016; 42(2):125-32. DOI: 10.1055/s-0035-1569068. View

Blasko E, Brooks A, Ho E, Wu J, Zhao X, Subramanyam B . Hepatocyte clearance and pharmacokinetics of recombinant factor IX glycosylation variants. Biochem Biophys Res Commun. 2013; 440(4):485-9. DOI: 10.1016/j.bbrc.2013.09.001. View

Petersen L, Elm T, Ezban M, Krogh T, Karpf D, Steino A . Plasma elimination kinetics for factor VII are independent of its activation to factor VIIa and complex formation with plasma inhibitors. Thromb Haemost. 2009; 101(5):818-26. View

10.

Lodge J, Jonas S, Oussoultzoglou E, Malago M, Jayr C, Cherqui D . Recombinant coagulation factor VIIa in major liver resection: a randomized, placebo-controlled, double-blind clinical trial. Anesthesiology. 2005; 102(2):269-75. DOI: 10.1097/00000542-200502000-00006. View

11.

Gupta S, Shukla P . Glycosylation control technologies for recombinant therapeutic proteins. Appl Microbiol Biotechnol. 2018; 102(24):10457-10468. DOI: 10.1007/s00253-018-9430-6. View

12.

Zhou Q, Qiu H . The Mechanistic Impact of N-Glycosylation on Stability, Pharmacokinetics, and Immunogenicity of Therapeutic Proteins. J Pharm Sci. 2018; 108(4):1366-1377. DOI: 10.1016/j.xphs.2018.11.029. View

13.

Pihusch M, Bacigalupo A, Szer J, von Depka Prondzinski M, Gaspar-Blaudschun B, Hyveled L . Recombinant activated factor VII in treatment of bleeding complications following hematopoietic stem cell transplantation. J Thromb Haemost. 2005; 3(9):1935-44. DOI: 10.1111/j.1538-7836.2005.01523.x. View

14.

Brenner B, Wiis J . Experience with recombinant-activated factor VII in 30 patients with congenital factor VII deficiency. Hematology. 2007; 12(1):55-62. DOI: 10.1080/10245330601111573. View

15.

OHara P, Grant F, Haldeman B, Gray C, Insley M, Hagen F . Nucleotide sequence of the gene coding for human factor VII, a vitamin K-dependent protein participating in blood coagulation. Proc Natl Acad Sci U S A. 1987; 84(15):5158-62. PMC: 298813. DOI: 10.1073/pnas.84.15.5158. View

16.

. Effect of early tranexamic acid administration on mortality, hysterectomy, and other morbidities in women with post-partum haemorrhage (WOMAN): an international, randomised, double-blind, placebo-controlled trial. Lancet. 2017; 389(10084):2105-2116. PMC: 5446563. DOI: 10.1016/S0140-6736(17)30638-4. View

17.

Narayan R, Maas A, Marshall L, Servadei F, Skolnick B, Tillinger M . Recombinant factor VIIA in traumatic intracerebral hemorrhage: results of a dose-escalation clinical trial. Neurosurgery. 2008; 62(4):776-86. DOI: 10.1227/01.neu.0000316898.78371.74. View

18.

Bosch J, Thabut D, Bendtsen F, DAmico G, Albillos A, Abraldes J . Recombinant factor VIIa for upper gastrointestinal bleeding in patients with cirrhosis: a randomized, double-blind trial. Gastroenterology. 2004; 127(4):1123-30. DOI: 10.1053/j.gastro.2004.07.015. View

19.

Rhys Thomas G, Dutton R, Hemlock B, Stein D, Hyder M, Shere-Wolfe R . Thromboembolic complications associated with factor VIIa administration. J Trauma. 2007; 62(3):564-9. DOI: 10.1097/TA.0b013e318031afc2. View

20.

Shakur H, Roberts I, Bautista R, Caballero J, Coats T, Dewan Y . Effects of tranexamic acid on death, vascular occlusive events, and blood transfusion in trauma patients with significant haemorrhage (CRASH-2): a randomised, placebo-controlled trial. Lancet. 2010; 376(9734):23-32. DOI: 10.1016/S0140-6736(10)60835-5. View