Rational design of a fully active, long-acting PEGylated factor VIII for hemophilia A treatment

Mei, Baisong; Pan, Clark Q.; Jiang, Haiyan; Tjandra, Hendri; Strauss, Jonathan B.; Chen, Yaoqi; Liu, Tongyao; Zhang, Xin; Severs, Joanne C.; Newgren, Jim; Chen, Jianmin; Gu, Jianming; Subramanyam, Babu; Fournel, Michael A.; Pierce, Glenn F.; Murphy, John E.

doi:10.1182/blood-2009-11-254755

Cited by 219 publications

(240 citation statements)

References 42 publications

(49 reference statements)

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“…The t 1/2 of recombinant FVIII or recombinant human B-domain deleted FVIII infused into mice is significantly shorter in the absence than presence of VWF (18 minutes vs 5.9-7.6 hours, respectively). [24][25][26] We confirm this relationship and demonstrate that a minimal, N-terminal VWF fragment comprising D9D3 domains (S764-P1247) is sufficient to stabilize endogenous FVIII in Vwf 2/2 mice. Extending the circulatory lifetime of D9D3 by Fc fusion markedly prolongs endogenous FVIII survival in Vwf 2/2 mice but is insufficient to extend the FVIII t 1/2 in HA mice.…”

Section: Discussionsupporting

confidence: 67%

“…21,22 Several approaches have attempted to engineer FVIII with an extended plasma half-life (t 1/2 ). [23][24][25][26] Recombinant human B-domain deleted FVIII infused into a C57BL/6 mouse has a similar t 1/2 (4.3 hours) to that of endogenous murine VWF (;4.5 hours). 25,27,28 Selective conjugation of polyethyleneglycol (PEG) 24,26 or fusion of FVIII to the Fc domain of immunoglobulin G (IgG) 25 increases FVIII plasma t 1/2 by ;1.5-to 2-fold in mice [24][25][26] and humans.…”

mentioning

confidence: 97%

“…[23][24][25][26] Recombinant human B-domain deleted FVIII infused into a C57BL/6 mouse has a similar t 1/2 (4.3 hours) to that of endogenous murine VWF (;4.5 hours). 25,27,28 Selective conjugation of polyethyleneglycol (PEG) 24,26 or fusion of FVIII to the Fc domain of immunoglobulin G (IgG) 25 increases FVIII plasma t 1/2 by ;1.5-to 2-fold in mice [24][25][26] and humans. 29 Here, we report the effect of a set of VWF fragments on FVIII binding in vitro and survival in vivo and demonstrate a markedly extended plasma t 1/2 for an Fc fusion to a 484-amino acid N-terminal VWF fragment, with a concomitant increase in plasma FVIII levels in VWF-deficient but not HA mice.…”

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confidence: 97%

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A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

Yee

Gildersleeve

et al. 2014

Blood

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• The D9D3 domains of VWF are sufficient to stabilize FVIII in vivo.• The prolongation of VWF D9D3 survival in vivo by Fc fusion elevates FVIII levels in the setting of VWF but not FVIII deficiency.Plasma factor VIII (FVIII) and von Willebrand factor (VWF) circulate together as a complex. We identify VWF fragments sufficient for FVIII stabilization in vivo and show that hepatic expression of the VWF D9D3 domains (S764-P1247), either as a monomer or a dimer, is sufficient to raise FVIII levels in Vwf 2/2 mice from a baseline of ∼5% to 10%, to ∼50% to 100%. These results demonstrate that a fragment containing only ∼20% of the VWF sequence is sufficient to support FVIII stability in vivo. Expression of the VWF D9D3 fragment fused at its C terminus to the Fc segment of immunoglobulin G1 results in markedly enhanced survival in the circulation (t 1/2 > 7 days), concomitant with elevated plasma FVIII levels (>25% at 7 days) in Vwf 2/2 mice. Although the VWF D9D3-Fc chimera also exhibits markedly prolonged survival when transfused into FVIII-deficient mice, the cotransfused FVIII is rapidly cleared. Kinetic binding studies show that VWF propeptide processing of VWF D9D3 fragments is required for optimal FVIII affinity. The reduced affinity of VWF D9D3 and VWF D9D3-Fc for FVIII suggests that the shortened FVIII survival in FVIII-deficient mice transfused with FVIII and VWF D9D3/D9D3-Fc is due to ineffective competition of these fragments with endogenous VWF for FVIII binding. (Blood. 2014; 124(3):445-452)

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Section: Discussionsupporting

confidence: 67%

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confidence: 97%

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confidence: 97%

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A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

Yee

Gildersleeve

et al. 2014

Blood

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“…Subsequent studies in haemophilic mice demonstrated a significantly extended half-life with di-PEGylated FVIII molecules and excellent in vivo efficacy compared with traditional BDD-FVIII (Fig. 2) [14]. A study in humans is currently ongoing.…”

Section: Chemical Conjugation To Prolong Protein Half-lifementioning

confidence: 97%

The role of natural VWF/FVIII complex concentrates in contemporary haemophilia care: a guideline for the next decade

Mannucci

2012

Haemophilia

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Summary. Current treatment for haemophilia provides excellent efficacy and safety albeit with a number of unresolved issues. The development of inhibitors following treatment with factor VIII (FVIII) is the most challenging complication of haemophilia and bears the highest economic burden for a chronic disease. Moreover, prophylactic therapy for haemophilia requires repeated infusions of FVIII, frequently as often as two or three times weekly, which can impact greatly on patientsÕ daily lives. As considerable scope remains for further advancements in the management of this condition, the primary focus of this paper relates to issues regarding current treatment and strategies in place to resolve the various issues. For countries approaching access to replacement therapy, it is important to know whether or not plasma-derived and recombinant products are associated with different risks of inhibitor development in previously untreated patients with severe haemophilia. The ongoing international SIPPET study is expected to provide an answer to this clinical dilemma. Methods under investigation to prolong the half-life of factor concentrates offer new hope to reduce the burden of prophylaxis for patients with haemophilia, with early results suggesting greater benefits with FIX.

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“…Mei et al (2006) showed that rFVIII expression in this cell line is 8 to 30-fold higher than the one obtained in other cell lines, such as HEK293 and BHK21 cells. 58 Bayer HealthCare is using this cell line for the production of Bay 94-9027 (damoctacog alfa pegol), a PEGylated recombinant factor VIII, 40,58 which is currently in a phase III clinical trial (clinicaltrials.gov).…”

Section: Recombinant Factor VIII Productsmentioning

confidence: 99%

Production of recombinant coagulation factors: Are humans the best host cells?

2017

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The main treatment option for Hemophilia A/B patients involves the administration of recombinant coagulation factors on-demand or in a prophylactic approach. Despite the safety and efficacy of this replacement therapy, the development of antibodies against the coagulation factor infused, which neutralize the procoagulant activity, is a severe complication. The production of recombinant coagulation factors in human cell lines is an efficient approach to avoid such complication. Human cell lines can produce recombinant proteins with post translation modifications more similar to their natural counterpart, reducing potential immunogenic reactions. This review provides a brief overview of the most important characteristics of recombinant FVIII and FIX products available on the market and the improvements that have recently been achieved by the production using human cell lines.

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Rational design of a fully active, long-acting PEGylated factor VIII for hemophilia A treatment

Cited by 219 publications

References 42 publications

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

The role of natural VWF/FVIII complex concentrates in contemporary haemophilia care: a guideline for the next decade

Production of recombinant coagulation factors: Are humans the best host cells?

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