Abstract
A novel high-affinity inhibitor of tumor necrosis factor (TNF) is described, which is created by the fusion of the extracellular domains of TNF-binding protein 1 (TBP-1) to both the α and β chains of an inactive version of the heterodimeric protein hormone, human chorionic gonadotropin. The resulting molecule, termed TNF-soluble high-affinity receptor complex (SHARC), self-assembles into a heterodimeric protein containing two functional TBP-1 moieties. The TNF-SHARC is a potent inhibitor of TNF-α bioactivity in vitro and has a prolonged pharmacokinetic profile compared with monomeric TBP-1 in vivo. Consistent with the long half-life, the duration of action in an lipopolysaccharide-mediated proinflammatory mouse model is prolonged similarly. In a collagen-induced arthritis mouse model, this molecule demonstrates improved efficacy over monomeric TBP-1. Based on these results, we demonstrated that inactivated heterodimeric protein hormones are flexible and efficient scaffolds for the creation of soluble high-affinity receptor complexes.
Footnotes
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.107.119875.
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ABBREVIATIONS: TNF, tumor necrosis factor; TBP, TNF-binding protein; SHARC, soluble high-affinity receptor complex; TNFR, TNF receptor; LPS, lipopolysaccharide; Fc, constant fragment; PAGE, polyacrylamide gel electrophoresis; hCG, human chorionic gonadotropin; CHO, Chinese hamster ovary; LH, luteinizing hormone; ELISA, enzyme-linked immunosorbent assay; Cu-IMAC, copper-immobilized metal-affinity chromatography; PBS, phosphate-buffered saline; DMEM, Dulbecco's modified Eagle's medium.
- Received January 11, 2007.
- Accepted May 9, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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