Pharmacokinetics Versus In Vitro Antiproliferative Potency to Design a Novel Hyperglycosylated hIFN-α2 Biobetter

AGUSTINA GUGLIOTTA; MARIA JESÚS LEOPOLD; EDUARDO MUFARREGE; MARINA ECHEVERRIGARAY; RICARDO KRATJE; NATALIA CEAGLIO; MARCOS OGGERO EBERHARDT

PHARMACEUTICAL RESEARCH

SPRINGER/PLENUM PUBLISHERS

Año: 2021

0724-8741

Purpose IFN4N is a glycoengineered version of recombinant human interferon alpha 2 (rhIFN-α2) that was modified to exhibit four N-glycosylation sites. It shows reduced in vitro specific biological activity (SBA) mainly due to R23 mutation by N23. However, it has improved pharmacokinetics and led to a high in vivo antitumor activity in mice. In order to prepare a new IFN-based biobetter, this work compares the influence of glycosylation (affecting pharmacokinetics) with the in vitro antiproliferativeSBA on the in vivo efficacy. Methods Based on IFN4N, three groups of muteins were designed, produced, and characterized. Group A: variants with the same glycosylation degree (4N) but higher in vitro antiproliferative SBA (R23 restored); group B: muteins with higher glycosylation degree (5N) but similar in vitro antiproliferative activity; and group C: variants with improved glycosylation(5N and 6N) and in vitro antiproliferative bioactivity. Results Glycoengineering was successful for improving pharmacokinetics, and R23 restoration considerably increased in vitro antiproliferative activity of new muteins compared toIFN4N. Hyperglycosylation was able to improve the in vivo efficacy similarly to or even better than R23 restoration. Additionally, the highest glycosylated mutein exhibited the lowest immunogenicity. Conclusions Hyperglycosylation constitutes a successful strategyto prepare a novel IFN biobetter.