Formatted anti-tumor necrosis factor α VHH proteins derived from camelids show superior potency and targeting to inflamed joints in a murine model of collagen-induced arthritis

被引:234
作者
Coppieters, Ken
Dreier, Torsten
Silence, Karen
de Haard, Hans
Lauwereys, Marc
Casteels, Peter
Beirnaert, Els
Jonckheere, Heidi
de Wiele, Christophe Van
Staelens, Ludovicus
Hostens, Jeroen
Revets, Hilde
Remaut, Erik
Elewaut, Dirk
Rottiers, Pieter
机构
[1] Univ Ghent VIB, Dept Mol Biomed Res, Appl Mol Bacteriol Unit, B-9052 Zwijnaarde, Belgium
[2] Ablynx NV, Zwijnaarde, Belgium
[3] Ghent Univ Hosp, B-9000 Ghent, Belgium
[4] Free Univ Brussels, Brussels, Belgium
来源
ARTHRITIS AND RHEUMATISM | 2006年 / 54卷 / 06期
关键词
D O I
10.1002/art.21827
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Objective. The advent of tumor necrosis factor (TNF)-blocking drugs has provided rheumatologists with an effective, but highly expensive, treatment for the management of established rheumatoid arthritis (RA). Our aim was to explore preclinically the application of camelid anti-TNF VHH proteins, which are single-domain antigen binding (VHH) proteins homologous to human immunoglobulin V-H domains, as TNF antagonists in a mouse model of RA. Methods. Llamas were immunized with human and mouse TNF, and antagonistic anti-TNF VHH proteins were isolated and cloned for bacterial production. The resulting anti-TNF VHH proteins were recombinantly linked to yield bivalent mouse and human TNF-specific molecules. To increase the serum half-life and targeting properties, an anti-serum albumin anti-TNF VHH domain was incorporated into the bivalent molecules. The TNF-neutralizing potential was analyzed in vitro. Mouse TNF-specific molecules were tested in a therapeutic protocol in murine collagen-induced arthritis (CIA). Disease progression was evaluated by clinical scoring and histologic evaluation. Targeting properties were evaluated by Tc-99m labeling and gamma camera imaging. Results. The bivalent molecules were up to 500 times more potent than the monovalent molecules. The antagonistic potency of the anti-human TNF VHH proteins exceeded even that of the anti-TNF antibodies infliximab and adalimumab that are used clinically in RA. Incorporation of binding affinity for albumin into the anti-TNF VHH protein significantly prolonged its serum half-life and promoted its targeting to inflamed joints in the murine CIA model of RA. This might explain the excellent therapeutic efficacy observed in vivo. Conclusion. These data suggest that because of the flexibility of their format, camelid anti-TNF VHH proteins can be converted into potent therapeutic agents that can be produced and purified cost-effectively.
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页码:1856 / 1866
页数:11
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