Creation of an improved mutant TNF with TNFR1-selectivity and antagonistic activity by phage display technology.

Abstract

Tumor necrosis factor-alpha (TNF), which binds two types of TNF receptors (TNFR1 and TNFR2), regulates the onset and exacerbation of autoimmune diseases such as rheumatoid arthritis and Crohn's disease. In particular, TNFR1-mediated signals are predominantly related to the induction of inflammatory responses. We have previously generated a TNFR1-selective antagonistic TNF-mutant (mutTNF) and shown that mutTNF efficiently inhibits TNFR1-mediated bioactivity in vitro and attenuates inflammatory conditions in vivo. In this study, we aimed to improve the TNFR1-selectivity of mutTNF This was achieved by constructing a phage library displaying mutTNF-based variants, in which the amino acid residues at the predicted receptor binding sites were substituted to other amino acids. From this mutant TNF library, 20 candidate TNFR1-selective antagonists were isolated. Like mutTNF, all 20 candidates were found to have an inhibitory effect on TNFR1-mediated bioactivity. However, one of the mutants, N7, displayed significantly more than 40-fold greater TNFR1-selectivty than mutTNF. Therefore, N7 could be a promising anti-autoimmune agent that does not interfere with TNFR2-mediated signaling pathways.

• Authors: T Nomura, Y Abe, H Kamada, M Inoue, T Kawara, S Arita, T Furuya, K Minowa, Y Yoshioka, H Shibata, H Kayamuro, T Yamashita, K Nagano, T Yoshikawa, Y Mukai, S Nakagawa, S Tsunoda, Y Tsutsumi
• Journal: Die Pharmazie (Pharmazie) Vol. 65 Issue 2 Pg. 93-6 (Feb 2010) ISSN: 0031-7144 [Print] Germany
• PMID: 20225650 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Peptide Library
• Receptors, Tumor Necrosis Factor, Type I
• Receptors, Tumor Necrosis Factor, Type II
• Tumor Necrosis Factors

Topics

• Cell Line
• Cell Survival (drug effects)
• Fibroblasts (drug effects)
• Genetic Variation
• Humans
• Mutation
• Peptide Library
• Receptors, Tumor Necrosis Factor, Type I (antagonists & inhibitors, genetics)
• Receptors, Tumor Necrosis Factor, Type II (drug effects)
• Surface Plasmon Resonance
• Tumor Necrosis Factors (genetics, pharmacology)