Ensovibep, a novel trispecific DARPin candidate that protects against SARS-CoV-2 variants
Sylvia Rothenberger, Daniel L Hurdiss, Marcel Walser, Francesca Malvezzi, Jennifer Mayor, Sarah Ryter, Hector Moreno, Nicole Liechti, Andreas Bosshart, Chloe Iss, Valérie Calabro, Andreas Cornelius, Tanja Hospodarsch, Alexandra Neculcea, Thamar Looser, Anja Schlegel, Simon Fontaine, Denis Villemagne, Maria Paladino, Yvonne Kaufmann, Doris Schaible, Iris Schlegel, Dieter Schiegg, Christof Zitt, Gabriel Sigrist, Marcel Straumann, Feyza Sacarcelik, Julia Wolter, Marco Comby, Julia M Adler, Kathrin Eschke, Mariana Nascimento, Azza Abdelgawad, Achim D Gruber, Judith Bushe, Olivia Kershaw, Heyrhyoung Lyoo, Chunyan Wang, Wentao Li, Ieva Drulyte, Wenjuan Du, H Kaspar Binz, Rachel Herrup, Sabrina Lusvarghi, Sabari Nath Neerukonda, Russell Vassell, Wei Wang, Susanne Mangold, Christian Reichen, Filip Radom, Charles G Knutson, Kamal K Balavenkatraman, Krishnan Ramanathan, Seth Lewis, Randall Watson, Micha A Haeuptle, Alexander Zürcher, Keith M Dawson, Daniel Steiner, Carol D Weiss, Patrick Amstutz, Frank J M Van Kuppeveld, Michael T Stumpp, Berend-Jan Bosch, Olivier Engler, Jakob Trimpert
doi:10.1101/2021.02.03.429164
SARS-CoV-2 has infected millions of people globally and continues to undergo evolution. Emerging variants can be partially resistant to vaccine induced and therapeutic antibodies, emphasizing the urgent need for accessible, broad-spectrum therapeutics. Here, we report a comprehensive study of ensovibep, the first trispecific clinical DARPin candidate, that can simultaneously engage all three units of the spike protein trimer to potently inhibit ACE2 interaction, as revealed by structural analyses. The cooperative binding of the individual modules enables ensovibep to retain inhibitory potency against all frequent SARS-CoV-2 variants, including Omicron, as of December 2021. Moreover, viral passaging experiments show that ensovibep, when used as a single agent, can prevent development of escape mutations comparably to a cocktail of monoclonal antibodies (mAb). Finally, we demonstrate that the very high in vitro antiviral potency also translates into significant therapeutic protection and reduction of pathogenesis in Roborovski dwarf hamsters infected with either the SARS-CoV-2 wild-type or the Alpha variant. In this model, ensovibep prevents fatality and provides substantial protection equivalent to the standard of care mAb cocktail. These results support further clinical evaluation and indicate that ensovibep could be a valuable alternative to mAb cocktails and other treatments for COVID-19.
Supplementary Materials for Ensovibep Supplementary Figure 1: A-C
Supplementary Figure 4: Titration curves for ensovibep (MP0420) and its RBD-binding domains (i.e. R1, R2 and R3), REGN10933 and REGN10987 to determine IC50 neutralization potencies on multiple spike mutants or only for ensovibep (MP0420) on the variants, which are summarized in Figure 2. Reported is the mean +/− SEM (standard error of the mean).
Supplementary Figure 5: Overview of the experimental protocol for viral passaging: A patient SARS-CoV-2 isolate from early 2020 (1.5 ×10 6 pfu) was incubated in presence of increasing concentrations of DARPin candidate or antibody for 4 days on Vero E6 cells and virus-induced cytopathic effects (CPE) were determined by microscopy. For each DARPin and antibody condition, cultures showing significant cytopathic effect (≥20%) under the greatest selective pressure were selected and virus-containing supernatant collected to start a new culture passage on Vero E6 cells (bold circle), again under increasing concentrations of the corresponding DARPin candidate or antibody condition. Passaging of virus containing supernatant was continued in the same manner for a total of 4 passages.
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'abstract': '<jats:title>Abstract</jats:title><jats:p>SARS-CoV-2 has infected millions of people globally '
'and continues to undergo evolution. Emerging variants can be partially resistant to vaccine '
'induced and therapeutic antibodies, emphasizing the urgent need for accessible, '
'broad-spectrum therapeutics. Here, we report a comprehensive study of ensovibep, the first '
'trispecific clinical DARPin candidate, that can simultaneously engage all three units of the '
'spike protein trimer to potently inhibit ACE2 interaction, as revealed by structural '
'analyses. The cooperative binding of the individual modules enables ensovibep to retain '
'inhibitory potency against all frequent SARS-CoV-2 variants, including Omicron, as of '
'December 2021. Moreover, viral passaging experiments show that ensovibep, when used as a '
'single agent, can prevent development of escape mutations comparably to a cocktail of '
'monoclonal antibodies (mAb). Finally, we demonstrate that the very high in vitro antiviral '
'potency also translates into significant therapeutic protection and reduction of pathogenesis '
'in Roborovski dwarf hamsters infected with either the SARS-CoV-2 wild-type or the Alpha '
'variant. In this model, ensovibep prevents fatality and provides substantial protection '
'equivalent to the standard of care mAb cocktail. These results support further clinical '
'evaluation and indicate that ensovibep could be a valuable alternative to mAb cocktails and '
'other treatments for COVID-19.</jats:p>',
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