PB1102 - Covalently linked complexes of coagulation factor VIII and von Willebrand factor fragment display increased stability and half-life in animal models and retain their function.

Tuesday, June 25, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Presenting Author(s)

BS

Barbara Solecka-Witulska

Scientist
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany

Co-Author(s)

AP

Anna Postępska-Igielska

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
AS

Anna Svirina

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
KD

Katja Derkow

Scientist
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany
UN

Ulrike Noller

Research associate
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
JW

Jessica Wernet

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
TH

Torsten Heidenreich

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
KR

Kirstin Rau

Technical Assistant
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany
IF

Inga Frey

Research associate
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
JI

Jennifer Ivlev

Research associate
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
CW

Cathleen Wegmann

Research associate
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
JZ

Jana Zell

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
AZ

Antra Zeltina

Scientist
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
AB

Anja Brehm

Scientist
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany
SW

Silvio Wuschko

Senior Director Pharmacology and Toxicology
Octapharma Pharmazeutika Produktionsges.m.b.H., Vienna, Austria
Wien, Wien, Austria
SW

Sonja Werwitzke

Deputy Director of Central Hematology and Hemostasis Laboratory
Clinic of Haematology, Haemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany
Hannover, Niedersachsen, Germany
Andreas Tiede, MD PhD

Professor of Hemostasis and Thrombosis
Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany
Hannover, Niedersachsen, Germany
TG

Thomas Güttler

Head of recombinant R&D
Octapharma Biopharmaceuticals GmbH, Recombinant R&D, Heidelberg, Germany
Heidelberg, Baden-Wurttemberg, Germany
GK

Guido Kohla

Head of molecular biochemistry
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany
CK

Christoph Kannicht

Scientific Advisor & Portfolio Strategy R&D
Octapharma Biopharmaceuticals GmbH, Molecular Biochemistry, Berlin, Germany
Berlin, Berlin, Germany

Background: The frequent intravenous administration of coagulation FVIII (FVIII) in Haemophilia A replacement therapy represents major burden for patients. Von Willebrand factor (VWF) and VWF-fragments are known to have a stabilizing effect on FVIII in circulation. We tested if covalently linked complexes of FVIII and VWF fragment engineered for increased O-glycosylation levels (OCTA12) will result in greater stability and improved pharmacokinetic (PK) properties.

Aims: To develop a stabilized version of FVIII with improved physicochemical and pharmacokinetic properties

Methods: We used a novel approach to covalently dimerize FVIII and OCTA12. This was followed by a stepwise incorporation of half-life prolonging moieties. First, the effect of insertion of the VWF-derived O-glycosylated sequence repeats (abbreviated as “EP-extension peptides”) into linker regions and on the C-terminus was evaluated. Second, we tested if FVIII-OCTA12 heterodimers fused to an albumin-binding single-domain (VHH) antibody display additional improvement. The well-expressing and active heterodimer variants were purified, and PK properties of selected constructs were tested in Haemophilia A (HemA) mouse and non-human primate (NHP) models. Functionality was assessed with the thrombin generation assay (TGA) and through a HemA Tail-Clip study.

Results: The combination of covalent heterodimerization with OCTA12 and insertion of EPs into linker regions resulted in significant half-life prolongation of FVIII (between 3-4-fold) in HemA mouse model. In the NHP model, the half-life of the one representative heterodimer candidate was 26±9 h, this could be further increased to 33±5 h by additional fusion of an albumin binding VHH. All constructs showed to effectively induce thrombin generation in the TGA assay (peak and endogenous thrombin potential) with slightly delayed time to peak. The in vivo Tail-Cut study in HemA mice showed that the tested constructs were able to effectively mediate blood clotting.

Conclusion(s): The covalent FVIII-OCTA12 complexes demonstrate preferable PK properties and represent promising future drug candidates.