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.
Footnotes
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ABBREVIATIONS:
- IL-10
- interleukin 10
- IL-10R
- IL-10 receptor
- mIL-10
- native murine IL-10
- PEG-mIL-10
- PEGylated murine IL-10
- hIL-10
- humanized IL-10
- hαhIL-10
- humanized anti-human IL-10 12G8 monoclonal antibody
- DOL
- degree of labeling
- SEC
- size exclusion chromatography
- HPLC
- high-performance liquid chromatography
- ELISA
- enzyme-linked immunosorbent assay
- PBS
- phosphate-buffered saline buffer
- FcRn
- neonatal Fc receptor
- mFcRn
- murine FcRn
- GST
- glutathione transferase
- mαmIL-10
- mouse anti-mouse IL-10 11D8 monoclonal antibody
- PK
- pharmacokinetic
- FELA
- fluorescence-emission-linked assay
- CT
- computer tomography
- PAGE
- polyacrylamide gel electrophoresis
- bis-Tris
- 2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol
- HMW
- high molecular weight
- VEGF
- vascular endothelial growth factor
- TNF-α
- tumor necrosis factor-α.
- Received August 26, 2011.
- Accepted November 14, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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