Functionalization of Tumor Necrosis Factor-α Using Phage Display Technique and PEGylation Improves Its Antitumor Therapeutic Window

• Published in: Clinical cancer research 2004-12, Vol.10 (24), p.8293-8300
• Main Authors: SHIBATA, Hiroko, YOSHIOKA, Yasuo, NAKAGAWA, Shinsaku, HAYAKAWA, Takao, NAGATA, Satoshi, YAMAGATA, Yuriko, MAYUMI, Tadanori, KAMADA, Haruhiko, TSUTSUMI, Yasuo, IKEMIZU, Shinji, KOBAYASHI, Kyoko, YAMAMOTO, Yoko, MUKAI, Yohei, OKAMOTO, Takayuki, TANIAI, Madoka, KAWAMURA, Maki, ABE, Yasuhiro
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic agents; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Biological and medical sciences; Female; Fibrosarcoma - drug therapy; Fibrosarcoma - metabolism; Lysine - chemistry; Medical sciences; Mice; Mice, Inbred BALB C; Models, Molecular; Mutation - genetics; Peptide Library; Pharmacology. Drug treatments; Polyethylene Glycols - chemistry; Polyethylene Glycols - metabolism; Polyethylene Glycols - pharmacology; Receptors, Tumor Necrosis Factor, Type I - chemistry; Receptors, Tumor Necrosis Factor, Type I - metabolism; Sarcoma, Experimental - drug therapy; Sarcoma, Experimental - metabolism; Structure-Activity Relationship; Tumor Necrosis Factor-alpha - chemistry; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-04-0770

Purpose: In this study, the optimization of antitumor therapy with tumor necrosis factor-α (TNF-α) was attempted. Experimental Design: Using the phage display technique, we created a lysine-deficient mutant TNF-α (mTNF-K90R). This mutant had higher affinities to both TNF receptors, despite reports that certain lysine residues play important roles in trimer formation and receptor binding. Results: The mTNF-K90R showed an in vivo therapeutic window that was 13-fold higher than that of the wild-type TNF-α (wTNF-α). This was due to the synergistic effect of its 6-fold stronger in vitro bioactivity and its 2-fold longer plasma half-life derived from its surface negative potential. The reason why the mTNF-K90R showed a higher bioactivity was understood by a molecular modeling analysis of the complex between the wTNF-α and TNF receptor-I. The mTNF-K90R, which was site-specifically mono-PEGylated at the NH 2 terminus (sp-PEG-mTNF-K90R), had a higher in vitro bioactivity and considerably longer plasma half-life than the wTNF-α, whereas the randomly mono-PEGylated wTNF-α had 6% of the bioactivity of the wTNF-α. With regard to effectiveness and safety, the in vivo antitumor therapeutic window of the sp-PEG-mTNF-K90R was 60-fold wider than that of the wTNF-α. Conclusions: These results indicated that this functionalized TNF-α may be useful not only as an antitumor agent but also as a selective enhancer of vascular permeability in tumors for improving antitumor chemotherapy.