Characterization of a HIR-Fab-IDS, Novel Iduronate 2-Sulfatase Fusion Protein for the Treatment of Neuropathic Mucopolysaccharidosis Type II (Hunter Syndrome)

• Published in: BioDrugs : clinical immunotherapeutics, biopharmaceuticals, and gene therapy biopharmaceuticals, and gene therapy, 2023-05, Vol.37 (3), p.375-395
• Main Authors: Gusarova, Valentina D., Smolov, Maxim A., Lyagoskin, Ivan V., Degterev, Maksim B., Rechetnik, Elizaveta V., Rodionov, Alexander V., Pantyushenko, Marina S., Shukurov, Rahim R.
• Format: Article
• Language: English
• Subjects: Affinity; Amino acid sequence; Animals; Antibodies; Biomedical and Life Sciences; Biomedicine; Blood-brain barrier; Cancer Research; Central nervous system; Disease; Endothelial cells; Endothelial Cells - metabolism; Enzymatic activity; Enzyme-linked immunosorbent assay; Enzymes; Fibroblasts; Fusion protein; Genetic disorders; Glycosaminoglycans; Glycosaminoglycans - metabolism; Glycosaminoglycans - therapeutic use; Glycosylation; Humans; Iduronic Acid; Insulin; Insulin receptors; Internalization; Intravenous administration; Investigations; Lysosomal storage diseases; Macaca fascicularis - metabolism; Mass spectrometry; Mass spectroscopy; Metabolic disorders; Mice; Molecular Medicine; Monoclonal antibodies; Mucopolysaccharidosis; Mucopolysaccharidosis II - drug therapy; Nervous system; Original Research Article; Peptides; Pharmaceuticals; Pharmacotherapy; Post-translation; Proteins; Reagents; Receptor, Insulin; Recombinant Proteins - therapeutic use; Scientific imaging; Statistical analysis; Surface plasmon resonance; United States
• ISSN: 1173-8804; 1179-190X
• DOI: 10.1007/s40259-023-00590-w

Background Mucopolysaccharidosis type II is a severe lysosomal storage disease caused by deficient activity of the enzyme iduronate-2-sulfatase. The only medicinal product approved by the US Food and Drug Administration for enzyme replacement therapy, recombinant iduronate-2-sulfatase (idursulfase, Elaprase ® ), is a large molecule that is not able to cross the blood–brain barrier and neutralize progressive damage of the central nervous system caused by the accumulation of glycosaminoglycans. Novel chimeric protein HIR-Fab-IDS is an anti-human insulin receptor Fab fragment fused to recombinant modified iduronate-2-sulfatase. This modification provides a highly selective interaction with the human insulin receptor, which leads to the HIR-Fab-IDS crossing the blood–brain barrier owing to internalization of the hybrid molecule by transcytosis into endothelial cells adjacent to the nervous system by the principle of a ‘molecular Trojan horse’. Objectives In this work, the physicochemical and biological characterization of a blood–brain barrier-penetrating fusion protein, HIR-Fab-IDS, is carried out. HIR-Fab-IDS consists of an anti-human insulin receptor Fab fragment fused to recombinant iduronate-2-sulfatase. Methods Comprehensive analytical characterization utilizing modern techniques (including surface plasmon resonance and mass spectrometry) was performed using preclinical and clinical batches of HIR-Fab-IDS. Critical quality parameters that determine the therapeutic effect of iduronate-2-sulfatase, as well as IDS enzymatic activity and in vitro cell uptake activity were evaluated in comparison with the marketed IDS product Elaprase ® (IDS RP). In vivo efficiency of HIR-Fab-IDS in reversing mucopolysaccharidosis type II pathology in IDS-deficient mice was also investigated. The affinity of the chimeric molecule for the INSR was also determined by both an enzyme-linked immunosorbent assay and surface plasmon resonance. We also compared the distribution of 125 I-radiolabeled HIR-Fab-IDS and IDS RP in the tissues and brain of cynomolgus monkeys after intravenous administration. Results The HIR-Fab-IDS primary structure investigation showed no significant post-translational modifications that could affect IDS activity, except for the formylglycine content, which was significantly higher for HIR-Fab-IDS compared with that for IDS RP (~ 76.5 vs ~ 67.7%). Because of this fact, the specific enzyme activity of HIR-Fab-IDS was slightly higher than that of IDS RP (~ 2