title: Effects of PEGylation and immune complex formation on the pharmacokinetics and biodistribution of recombinant interleukin 10 in mice.
abstract: Interleukin 10 (IL-10) is a potent cytokine homodimer with multiple immunoregulatory functions. Here, we have characterized the effects of PEGylation and formation of human IL-10 (hIL-10)/humanized anti-human IL-10 (hαhIL-10) immune complexes in the pharmacokinetics, biodistribution, and biotransformation of IL-10 in mice. To assess the fate of native, PEGylated, and antibody-bound IL-10; we implemented an analytical set of fluorescence emission-linked assays. Plasma size exclusion chromatography analysis indicated that fluoro-labeled native and PEGylated murine IL-10 (PEG-mIL-10) are stable in the circulation. PEGylation of IL-10 resulted in a 21-fold increased exposure, 2.7-fold increase in half-life, and 20-fold reduction in clearance. Kidney is the major organ of disposition for both native and PEGylated mIL-10 with renal uptake directly related to systemic clearance. The fluorescence signal in the kidneys reached tissue/blood ratios up to 150 and 20 for native and PEG-mIL-10, respectively. hIL-10/hαhIL-10 immune complexes are detectable in the circulation without evidence of unbound or degraded hIL-10. The exposure of hIL-10 present in immune complexes versus that of hIL-10 alone increased from 0.53 to 11.28 μg · day/ml, with a half-life of 1.16 days and a 23-fold reduction in clearance. Unlike hIL-10 alone, antibody-bound hIL-10 was targeted mainly to the liver with minimal renal distribution. In addition, we found an 11-fold reduction (from 9.9 to 113 nM) in binding to the neonatal Fc receptor (FcRn) when the hαhIL10 antibody is conjugated to hIL-10. The potential changes in FcRn binding in vivo and increased liver uptake may explain the unique pharmacokinetic properties of hIL-10/hαhIL-10 immune complexes.
authors.affiliation: Biologics Discovery DMPK and Bioanalytics Department, Merck Research Laboratories, Merck Sharp and Dohme Corp., Palo Alto, California, USA.
authors.forename: Hamsell M
authors.initials: HM
authors.lastname: Alvarez
authors.forename: On-Yee
authors.initials: OY
authors.lastname: So
authors.forename: SuChun
authors.initials: S
authors.lastname: Hsieh
authors.forename: Natasha
authors.initials: N
authors.lastname: Shinsky-Bjorde
authors.forename: Huiping
authors.initials: H
authors.lastname: Ma
authors.forename: Yaoli
authors.initials: Y
authors.lastname: Song
authors.forename: Yinuo
authors.initials: Y
authors.lastname: Pang
authors.forename: Melinda
authors.initials: M
authors.lastname: Marian
authors.forename: Enrique
authors.initials: E
authors.lastname: Escandón
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics*; D000936:Antigen-Antibody Complex / Q000032:analysis; D000937:Antigen-Antibody Reactions; D005260:Female; D006207:Half-Life; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D016753:Interleukin-10 / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics*; D007668:Kidney / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D008657:Metabolic Clearance Rate; D051379:Mice; D008815:Mice, Inbred Strains; D011092:Polyethylene Glycols / Q000737:chemistry*; D011961:Receptors, Fc / Q000378:metabolism; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D014018:Tissue Distribution
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D000936:Antigen-Antibody Complex; D015395:Histocompatibility Antigens Class I; C508609:IL10 protein, human; C556062:IL10 protein, mouse; D011961:Receptors, Fc; D011994:Recombinant Proteins; D016753:Interleukin-10; D011092:Polyethylene Glycols; C100688:Fc receptor, neonatal