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Thromb Haemost 2008; 100(05): 920-928
DOI: 10.1160/TH08-04-0268
New Technologies, Diagnostic Tools and Drugs
Schattauer GmbH

Generation and biochemical characterization of glycoPEGylated factor VIIa derivatives

Henning R. Stennicke
1   Biopharmaceuticals Research Unit, Novo Nordisk, Novo Nordisk Park, Maaloev, Denmark
,
Henrik Østergaard
1   Biopharmaceuticals Research Unit, Novo Nordisk, Novo Nordisk Park, Maaloev, Denmark
,
Robert J. Bayer
2   Neose Technologies, Horsham, Pennsylvania, USA
,
Matt S. Kalo
2   Neose Technologies, Horsham, Pennsylvania, USA
,
Kyle Kinealy
2   Neose Technologies, Horsham, Pennsylvania, USA
,
Pernille K. Holm
1   Biopharmaceuticals Research Unit, Novo Nordisk, Novo Nordisk Park, Maaloev, Denmark
,
Brit B. Sørensen
1   Biopharmaceuticals Research Unit, Novo Nordisk, Novo Nordisk Park, Maaloev, Denmark
,
David Zopf
2   Neose Technologies, Horsham, Pennsylvania, USA
,
Søren E. Bjørn
1   Biopharmaceuticals Research Unit, Novo Nordisk, Novo Nordisk Park, Maaloev, Denmark
› Author Affiliations
Further Information

Publication History

Received 30 April 2008

Accepted after minor revision 08 August 2008

Publication Date:
22 November 2017 (online)

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Summary

Prophylaxis with 2–4 times weekly dosing of factor (F)VIII or FIX is established as an efficacious and safe treatment in haemophilia. Although prophylaxis is not readily available for the inhibitor patient,recent studies have demonstrated a reduction in bleeding episodes in inhibitor patients treated with daily infusions of FVIIa. In order to develop a treatment option comparable to prophylaxis with FVIII or FIX we looked to PEGylation which is an established method for prolonging the circulatory half-life of proteins. However, due to the numerous interactions of FVIIa with the cell surface,TF,FIX and FX there are limited options for unspecific chemical modification of FVIIa without loss of activity. Consequently, we explored the GlycoPEGylation™ technology for selective PEGylation of the two N-glycans in the FVIIa light chain and protease domain to generate seven specifically modified derivatives with PEG groups ranging from 2 to 40 kDa. These derivatives were evaluated in vitro for their ability to interact with small synthetic substrates as well as key molecules relevant to function in the coagulation pathway. The results demonstrate that modification of FVIIa using glycoPEGylation has only a very limited effect on the hydrolysis S-2288 and FX activation. However, the modification does to some extend alter the ability of FVIIa to interact with TF and more importantly, reduces the rate of ATIII inhibition by up to 50% which could allow for an extended active half-life in circulation.

Keywords

Long-acting - PEG - coagulation - serine protease - factor VIIa - prophylaxis
 

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