title: Conjugation to 10 kDa Linear PEG Extends Serum Half-Life and Preserves the Receptor-Binding Ability of mmTRAIL with Minimal Stimulation of PEG-Specific Antibodies.
abstract: The poor in vivo potencies of most therapeutic proteins might be attributed to their short serum half-lives. PEGylation is a well-established method and has been clinically proven to improve pharmacokinetics. mmTRAIL exhibited supercytotoxicity in a variety of tumor cells, but its serum half-life was less than 10 min in mice. Here, mmTRAIL-5K, mmTRAIL-10K, and mmTRAIL-20K were produced by N-terminus-specific PEGylation of mmTRAIL with 5, 10, or 20 kDa mPEG, respectively. The particle sizes of mmTRAIL-5K, mmTRAIL-10K, and mmTRAIL-20K were 9.09 ± 2.76, 12.62 ± 4.05, and 15.68 ± 4.95 nm, which were higher than the threshold (∼7 nm) of renal clearance. Accordingly, mmTRAIL-5K exhibited a serum half-life of 30 min only 3 times longer than that of mmTRAIL. However, both mmTRAIL-10K and mmTRAIL-20K exhibited similar serum half-lives ranging from 350 to 400 min, indicating that PEGylation with 10 or 20 kDa mPEG significantly improved the pharmacokinetics of mmTRAIL. However, death receptor binding of mmTRAIL-20K was reduced 5- to 8-fold, resulting in a 3-fold reduction of cytotoxicity. Additionally, repeated administration of mmTRAIL-20K elicited both mPEG-specific IgG and IgM antibody responses in rats. In contrast, the receptor binding and cytotoxicity of mmTRAIL-10K were similar to those of mmTRAIL. Repeated administration of mmTRAIL-10K did not obviously stimulate mPEG-specific antibody responses in rats and rhesus monkeys. Of the three PEGylated mmTRAIL analogues, mmTRAIL-10K exerted the greatest tumor suppression in mice bearing human tumor xenografts. These results demonstrated that conjugation of mmTRAIL to 10 kDa mPEG was better than that to 5 or 20 kDa mPEG for enhancing antitumor effects.
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Qianxue
authors.initials: Q
authors.lastname: Nie
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Dianlong
authors.initials: D
authors.lastname: Jia
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Yanru
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Fan
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Qiuxiao
authors.initials: Q
authors.lastname: Shi
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wan
authors.affiliation: Key Lab of Transplant Engineering and Immunology, MOH, West China Hospital, Sichuan University , Chengdu 610041, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Lu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics*; D045744:Cell Line, Tumor; D006207:Half-Life; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011092:Polyethylene Glycols / Q000737:chemistry*; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism*
chemical_list: D000906:Antibodies; D000970:Antineoplastic Agents; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D053221:TNF-Related Apoptosis-Inducing Ligand; D011092:Polyethylene Glycols
keywords: PEGylation; anti-PEG antibody; cancer targeted therapy; serum half-life; tumor necrosis factor-related apoptosis-inducing ligand