TY - JOUR
T1 - Sialic Acid-Engineered IL4-10 Fusion Protein is Bioactive and Rapidly Cleared from the Circulation
AU - Steen-Louws, Cristine
AU - Boross, Peter
AU - Prado, Judith
AU - Meeldijk, Jan
AU - Langenhorst, Jurgen B
AU - Huitema, Alwin D R
AU - den Hartog, Marcel T
AU - Boon, Louis
AU - Lafeber, Floris P J G
AU - Hack, C Erik
AU - Eijkelkamp, Niels
AU - Popov-Celeketic, Jelena
N1 - Funding Information:
We thank Helma van Doorn, Jules Teuwen, Joyce Tijhuis, Sabine Versteeg (UMC Utrecht), Anja van der Sar and Ivonne Dreumel (animal facility Utrecht University, the Netherlands) for technical assistance. Cristine Steen-Louws, Judith Prado, Erik Hack, Niels Eijkelkamp and Jelena Popov-Celeketic are shareholders of Synerkine Pharma BV, a recently founded company in which in which preclinical development and evaluation of IL4?10 fusion protein is enrolled. Cristine Steen-Louws, Peter Boross, Judith Prado, Alwin Huitema, Floris Lafeber, Erik Hack, Niels Eijkelkamp and Jelena Popov-Celeketic are co-inventors of the patent ?Modified therapeutic glycoprotein for local administration?. Erik Hack and Floris Lafeber are consultants for Synerkine Pharma BV. Jelena Popov-Celeketic is partly employed by Synerkine Pharma BV.
Publisher Copyright:
© 2019, The Author(s).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2019/12
Y1 - 2019/12
N2 - PURPOSE: Modulating sialylation of therapeutic glycoproteins may be used to influence their clearance and systemic exposure. We studied the effect of low and high sialylated IL4-10 fusion protein (IL4-10 FP) on in vitro and in vivo bioactivity and evaluated the effect of differential sialylation on pharmacokinetic parameters.METHODS: CHO cell lines producing low (IL4-10 FP lowSA) and high sialylated (IL4-10 FP highSA) fusion protein were generated. Bioactivity of the proteins was evaluated in an LPS-stimulated whole blood assay. Pharmacokinetics were studied in rats, analyzing plasma levels of IL4-10 FP upon intravenous injection. In vivo activity was assessed in an inflammatory pain mice model upon intrathecal injection.RESULTS: IL4-10 FP lowSA and IL4-10 FP highSA had similar potency in vitro. The pharmacokinetics study showed a 4-fold higher initial systemic clearance of IL4-10 FP lowSA, whereas the calculated half-life of both IL4-10 FP lowSA and IL4-10 FP highSA was 20.7 min. Finally, both IL4-10 FP glycoforms inhibited persistent inflammatory pain in mice to the same extent.CONCLUSIONS: Differential sialylation of IL4-10 fusion protein does not affect the in vitro and in vivo activity, but clearly results in a difference in systemic exposure. The rapid systemic clearance of low sialylated IL4-10 FP could be a favorable characteristic to minimize systemic exposure after administration in a local compartment.
AB - PURPOSE: Modulating sialylation of therapeutic glycoproteins may be used to influence their clearance and systemic exposure. We studied the effect of low and high sialylated IL4-10 fusion protein (IL4-10 FP) on in vitro and in vivo bioactivity and evaluated the effect of differential sialylation on pharmacokinetic parameters.METHODS: CHO cell lines producing low (IL4-10 FP lowSA) and high sialylated (IL4-10 FP highSA) fusion protein were generated. Bioactivity of the proteins was evaluated in an LPS-stimulated whole blood assay. Pharmacokinetics were studied in rats, analyzing plasma levels of IL4-10 FP upon intravenous injection. In vivo activity was assessed in an inflammatory pain mice model upon intrathecal injection.RESULTS: IL4-10 FP lowSA and IL4-10 FP highSA had similar potency in vitro. The pharmacokinetics study showed a 4-fold higher initial systemic clearance of IL4-10 FP lowSA, whereas the calculated half-life of both IL4-10 FP lowSA and IL4-10 FP highSA was 20.7 min. Finally, both IL4-10 FP glycoforms inhibited persistent inflammatory pain in mice to the same extent.CONCLUSIONS: Differential sialylation of IL4-10 fusion protein does not affect the in vitro and in vivo activity, but clearly results in a difference in systemic exposure. The rapid systemic clearance of low sialylated IL4-10 FP could be a favorable characteristic to minimize systemic exposure after administration in a local compartment.
KW - cytokines
KW - inflammation
KW - pharmacokinetics
KW - sialylation
KW - therapeutic protein
UR - http://www.scopus.com/inward/record.url?scp=85077034320&partnerID=8YFLogxK
U2 - 10.1007/s11095-019-2744-y
DO - 10.1007/s11095-019-2744-y
M3 - Article
C2 - 31879800
SN - 0724-8741
VL - 37
JO - Pharmaceutical Research
JF - Pharmaceutical Research
IS - 2
M1 - 17
ER -