Physicochemical and Biological Characterization of rhC1INH Expressed in CHO Cells.

Ekaterina Zubareva,Alexander Kazarov,Maria Zhiliaeva,Madina Iskakova,Rahim Shukurov,Ivan Lyagoskin,Vladimir Simonov,Anna Azarova,Ksenia Ulyanova,Maksim Smolov,Maksim Degterev

doi:10.3390/ph14111180

Abstract

The disfunction or deficiency of the C1 esterase inhibitor (C1INH) is associated with hereditary or acquired angioedema (HAE/AAE), a rare life-threatening condition characterized by swelling in the skin, respiratory and gastrointestinal tracts. The current treatment options may carry the risks of either viral infection (plasma-derived Berinert®) or immune reaction (human recombinant C1INH from rabbit milk, Ruconest®). This study describes the physicochemical and biological characterization of a novel recombinant human C1 esterase inhibitor (rhC1INH) from Chinese hamster ovary (CHO) cells for the treatment of hereditary angioedema compared to the marketed products Berinert® and Ruconest®. The mass spectrometry results of total deglycosylated rhC1INH revealed a protein with a molecular mass of 52,846 Da. Almost full sequence coverage (98.6%) by nanoLC-MS/MS peptide mapping was achieved. The purity and C1s inhibitory activity of rhC1INH from CHO cells are comparable with Ruconest®, although we found differences in charge isoforms distribution, intact mass values, and N-glycans profile. Comparison of the specific activity (IC50 value) of the rhC1INH with human C1 esterase inhibitor from blood serum showed similar inhibitory properties. These data allow us to conclude that the novel rhC1INH molecule could become a potential therapeutic option for patients with HAE/AAE.

Highlights

Hereditary angioedema (HAE) is a rare disease having an orphan status
This study describes the physicochemical and biological characterization of the novel recombinant human C1 esterase inhibitor (rhC1INH) from Chinese hamster ovary (CHO) cells
The product quality attributes of the rhC1INH were determined by employing state-of-the-art analytical methods

Summary

Introduction

Hereditary angioedema (HAE) is a rare disease having an orphan status. The study of HAE began in 1962 by Landerman and colleagues [1]. The biological role of C1 esterase inhibitor (C1 inhibitor) is to regulate two mutually dependent processes by inhibition of serine proteases responsible for vascular permeability and inflammatory response. The C1 inhibitor regulates the contact system, known as the kallikrein-kinin system. The activation of this system results in bradykinin generation. Contact system regulation occurs due to inhibition of the first complement component of the C1 macromolecule.

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Conclusion