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.
References
Baum, Antibody cocktail to SARS-CoV-2 spike protein prevents rapid mutational escape seen with individual antibodies, Science,
doi:10.1126/science.abd0831Berger Rentsch, Zimmer, A vesicular stomatitis virus replicon-based bioassay for the rapid and sensitive determination of multi-species type I interferon, PLoS One,
doi:10.1371/journal.pone.0025858Binz, High-affinity binders selected from designed ankyrin repeat protein libraries, Nat Biotechnol,
doi:10.1038/nbt962Cathcart, The dual function monoclonal antibodies VIR-7831 and VIR-7832 demonstrate potent in vitro and in vivo activity against SARS-CoV-2, bioRxiv,
doi:10.1101/2021.03.09.434607v1Cele, SARS-CoV-2 Omicron has extensive but incomplete escape of Pfizer BNT162b2 elicited neutralization and requires ACE2 for infection, medRxiv,
doi:10.1101/2021.12.08.21267417Choi, Proceedings of the ACM Conference on Bioinformatics, Computational Biology and Biomedicine
Cianfrocco, Wong, Youn, Wagner, The Practice and Experience in Advanced Research Computing
Cingolani, A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3, Fly (Austin),
doi:10.4161/fly.19695Copin, In vitro and in vivo preclinical studies predict REGEN-COV protection against emergence of viral escape in humans,
doi:10.1101/2021.03.10.434834Copin, In vitro and in vivo preclinical studies predict REGEN-COV protection against emergence of viral escape in humans, bioRxiv,
doi:10.1101/2021.03.10.434834v3Copin, The monoclonal antibody combination REGEN-COV protects against SARS-CoV-2 mutational escape in preclinical and human studies, Cell,
doi:10.1016/j.cell.2021.06.002Duvaud, the Swiss Bioinformatics Resource Portal, as designed by its users, Nucleic Acids Res,
doi:10.1093/nar/gkab225Fiedler, MP0250, a VEGF and HGF neutralizing DARPin((R)) molecule shows high anti-tumor efficacy in mouse xenograft and patient-derived tumor models, Oncotarget,
doi:10.18632/oncotarget.21738Friedrich, Preclinical characterization of AMG 330, a CD3/CD33-bispecific T-cell-engaging antibody with potential for treatment of acute myelogenous leukemia, Mol Cancer Ther,
doi:10.1158/1535-7163.MCT-13-0956Gobeil, Effect of natural mutations of SARS-CoV-2 on spike structure, conformation, and antigenicity, Science,
doi:10.1126/science.abi6226Goddard, UCSF ChimeraX: Meeting modern challenges in visualization and analysis, Protein Sci,
doi:10.1002/pro.3235Greaney, Complete Mapping of Mutations to the SARS-CoV-2 Spike Receptor-Binding Domain that Escape Antibody Recognition, Cell host & microbe,
doi:10.1016/j.chom.2020.11.007Gruber, Standardization of Reporting Criteria for Lung Pathology in SARS-CoV-2-infected Hamsters: What Matters?, Am J Respir Cell Mol Biol,
doi:10.1165/rcmb.2020-0280LEHoffmann, SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor, Cell,
doi:10.1016/j.cell.2020.02.052Jones, LY-CoV555, a rapidly isolated potent neutralizing antibody, provides protection in a non-human primate model of SARS-CoV-2 infection, bioRxiv,
doi:10.1101/2020.09.30.318972Kumari, Neuroinvasion and Encephalitis Following Intranasal Inoculation of SARS-CoV-2 in K18-hACE2 Mice, Viruses,
doi:10.3390/v13010132Laffeber, De Koning, Kanaar, Lebbink, Experimental Evidence for Enhanced Receptor Binding by Rapidly Spreading SARS-CoV-2 Variants, Journal of Molecular Biology,
doi:10.1016/j.jmb.2021.167058Laskowski, Swindells, LigPlot+: multiple ligand-protein interaction diagrams for drug discovery, J Chem Inf Model,
doi:10.1021/ci200227uLetko, Marzi, Munster, Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses, Nat Microbiol,
doi:10.1038/s41564-020-0688-yLiu, Identification of SARS-CoV-2 spike mutations that attenuate monoclonal and serum antibody neutralization, Cell host & microbe,
doi:10.1016/j.chom.2021.01.014Lu3, Peng1, Sterling4, Walsh, Structural impact on SARS SoV-2 spike protein by D614G substitution
Lusvarghi, Key substitutions in the spike protein of SARS-CoV-2 variants can predict resistance to monoclonal antibodies, but other substitutions can modify the effects, bioRxiv,
doi:10.1101/2021.07.16.452748Neerukonda, Establishment of a well-characterized SARS-CoV-2 lentiviral pseudovirus neutralization assay using 293T cells with stable expression of ACE2 and TMPRSS2, PLoS One,
doi:10.1371/journal.pone.0248348Nouailles, Temporal omics analysis in Syrian hamsters unravel cellular effector responses to moderate COVID-19, Nat Commun,
doi:10.1038/s41467-021-25030-7Osterrieder, Age-Dependent Progression of SARS-CoV-2 Infection in Syrian Hamsters, Viruses,
doi:10.3390/v12070779Pettersen, UCSF Chimera--a visualization system for exploratory research and analysis, J Comput Chem,
doi:10.1002/jcc.20084Pulliam, Increased risk of SARS-CoV-2 reinfection associated with emergence of the Omicron variant in South Africa, medRxiv,
doi:10.1101/2021.11.11.21266068Sim, SIFT web server: predicting effects of amino acid substitutions on proteins, Nucleic Acids Res,
doi:10.1093/nar/gks539Starr, Deep Mutational Scanning of SARS-CoV-2 Receptor Binding Domain Reveals Constraints on Folding and ACE2 Binding, Cell,
doi:10.1016/j.cell.2020.08.012Starr, Deep Mutational Scanning of SARS-CoV-2 Receptor Binding Domain Reveals Constraints on Folding and ACE2 Binding, Cell,
doi:10.1016/j.cell.2020.08.012Tegally, Emergence and rapid spread of a new severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) lineage with multiple spike mutations in South Africa, medRxiv,
doi:10.1101/2020.12.21.20248640Thomson, The circulating SARS-CoV-2 spike variant N439K maintains fitness while evading antibody-mediated immunity, bioRxiv,
doi:10.1101/2020.11.04.355842Torriani, Identification of Clotrimazole Derivatives as Specific Inhibitors of Arenavirus Fusion, J Virol,
doi:10.1128/JVI.01744-18Trimpert, The Roborovski Dwarf Hamster Is A Highly Susceptible Model for a Rapid and Fatal Course of SARS-CoV-2 Infection, Cell reports,
doi:10.1016/j.celrep.2020.108488Walls, Cryo-electron microscopy structure of a coronavirus spike glycoprotein trimer, Nature,
doi:10.1038/nature16988Walls, Tectonic conformational changes of a coronavirus spike glycoprotein promote membrane fusion, Proc Natl Acad Sci U S A,
doi:10.1073/pnas.1708727114Walser, Highly potent anti-SARS-CoV-2 multivalent DARPin therapeutic candidates
Walter, Hutter, Garaeva, Scherer, Zimmermann, Highly potent bispecific sybodies neutralize SARS-CoV-2
Wilm, LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cellpopulation heterogeneity from high-throughput sequencing datasets, Nucleic Acids Res,
doi:10.1093/nar/gks918Wollscheid, Mass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins, Nat Biotechnol,
doi:10.1038/nbt.1532Yi, Key residues of the receptor binding motif in the spike protein of SARS-CoV-2 that interact with ACE2 and neutralizing antibodies, Cell Mol Immunol,
doi:10.1038/s41423-020-0458-zZahradnik, SARS-CoV-2 RBD in vitro evolution follows contagious mutation spread, yet generates an able infection inhibitor, bioRxiv,
doi:10.1101/2021.01.06.425392v3Zheng, MotionCor2: anisotropic correction of beam-induced motion for improved cryoelectron microscopy, Nat Methods,
doi:10.1038/nmeth.4193Zivanov, New tools for automated high-resolution cryo-EM structure determination in RELION-3, Elife,
doi:10.7554/eLife.42166DOI record:
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