Multivalency transforms SARS-CoV-2 antibodies into broad and ultrapotent neutralizers

AbstractThe novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes Coronavirus Disease 2019 (COVID-19), has caused a global pandemic. Antibodies are powerful biotherapeutics to fight viral infections; however, discovery of the most potent and broadly acting clones can be lengthy. Here, we used the human apoferritin protomer as a modular subunit to drive oligomerization of antibody fragments and transform antibodies targeting SARS-CoV-2 into exceptionally potent neutralizers. Using this platform, half-maximal inhibitory concentration (IC50) values as low as 9 × 10−14 M were achieved as a result of up to 10,000-fold potency enhancements. Combination of three different antibody specificities and the fragment crystallizable (Fc) domain on a single multivalent molecule conferred the ability to overcome viral sequence variability together with outstanding potency and Ig-like in vivo bioavailability. This MULTi-specific, multi-Affinity antiBODY (Multabody; or MB) platform contributes a new class of medical countermeasures against COVID-19 and an efficient approach to rapidly deploy potent and broadly-acting therapeutics against infectious diseases of global health importance.One Sentence Summarymultimerization platform transforms antibodies emerging from discovery screens into potent neutralizers that can overcome SARS-CoV-2 sequence diversity.

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Multivalency transforms SARS-CoV-2 antibodies into ultrapotent neutralizers

Nature Communications ◽

10.1038/s41467-021-23825-2 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Edurne Rujas ◽

Iga Kucharska ◽

Yong Zi Tan ◽

Samir Benlekbir ◽

Hong Cui ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Inhibitory Concentration ◽

Viral Infections ◽

Antibody Fragments ◽

Rapid Evaluation ◽

Sequence Variability ◽

Viral Sequence ◽

Global Pandemic ◽

Antibody Specificities ◽

Binding Avidity

AbstractSARS-CoV-2, the virus responsible for COVID-19, has caused a global pandemic. Antibodies can be powerful biotherapeutics to fight viral infections. Here, we use the human apoferritin protomer as a modular subunit to drive oligomerization of antibody fragments and transform antibodies targeting SARS-CoV-2 into exceptionally potent neutralizers. Using this platform, half-maximal inhibitory concentration (IC50) values as low as 9 × 10−14 M are achieved as a result of up to 10,000-fold potency enhancements compared to corresponding IgGs. Combination of three different antibody specificities and the fragment crystallizable (Fc) domain on a single multivalent molecule conferred the ability to overcome viral sequence variability together with outstanding potency and IgG-like bioavailability. The MULTi-specific, multi-Affinity antiBODY (Multabody or MB) platform thus uniquely leverages binding avidity together with multi-specificity to deliver ultrapotent and broad neutralizers against SARS-CoV-2. The modularity of the platform also makes it relevant for rapid evaluation against other infectious diseases of global health importance. Neutralizing antibodies are a promising therapeutic for SARS-CoV-2.

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Non-Toxic Virucidal Macromolecules Show High Efficacy Against Influenza Virus Ex Vivo and In Vivo

10.1101/2020.03.18.996678 ◽

2020 ◽

Author(s):

Ozgun Kocabiyik ◽

Valeria Cagno ◽

Paulo Jacob Silva ◽

Yong Zhu ◽

Laura Sedano ◽

...

Keyword(s):

Broad Spectrum ◽

Viral Infections ◽

Ex Vivo ◽

Public Health Problem ◽

Major Public Health Problem ◽

New Class ◽

Influenza Strain ◽

Low Toxicity

AbstractInfluenza is one of the most widespread viral infections worldwide and represents a major public health problem. The risk that one of the next pandemics is caused by an influenza strain is very high. It is very important to develop broad-spectrum influenza antivirals to be ready for any possible vaccine shortcomings. Anti-influenza drugs are available but they are far from ideal. Arguably, an ideal antiviral should target conserved viral domains and be virucidal, i.e. irreversibly inhibit viral infectivity. Here, we describe a new class of broad-spectrum anti-influenza macromolecules that meets these criteria and displays exceedingly low toxicity. These compounds are based on a cyclodextrin core modified on its primary face with long hydrophobic linkers terminated in 6’sialyl-N-acetyllactosamine (6’SLN) or 3’SLN. SLN enables nanomolar inhibition of the viruses while the hydrophobic linkers confer irreversibility to the inhibition. The combination of these two properties allows for efficacy in vitro against several human or avian influenza strains, as well as against a 2009 pandemic influenza strain ex vivo. Importantly, we show that, in mice, the compounds provide therapeutic efficacy when administered 24h post-infection allowing 90% survival as opposed to no survival for the placebo and oseltamivir..

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Quantifying synergy in the bioassay-guided fractionation of natural product extracts

10.1101/2020.06.23.166686 ◽

2020 ◽

Cited By ~ 1

Author(s):

Micah Dettweiler ◽

Lewis Marquez ◽

Max Bao ◽

Cassandra L. Quave

Keyword(s):

Inhibitory Concentration ◽

Viral Infections ◽

Fractional Inhibitory Concentration ◽

Drug Mixtures ◽

Natural Substances ◽

Complex Interactions ◽

Bioassay Guided Fractionation ◽

Using Data ◽

Living Organisms

AbstractMixtures of drugs often have greater therapeutic value than any of their constituent drugs alone, and such combination therapies are widely used to treat diseases such as cancer, malaria, and viral infections. However, developing useful drug mixtures is challenging due to complex interactions between drugs. Natural substances can be fruitful sources of useful drug mixtures because secondary metabolites produced by living organisms do not often act in isolation in vivo. In order to facilitate the study of interactions within natural substances, a new analytical method to quantify interactions using data generated in the process of bioassay-guided fractionation is presented here: the extract fractional inhibitory concentration index (EFICI). The EFICI method uses the framework of Loewe additivity to calculate fractional inhibitory concentration values by which interactions can be determined for any combination of fractions that make up a parent extract. The EFICI method was applied to data on the bioassay-guided fractionation of Lechea mucronata and Schinus terebinthifolia for growth inhibition of the pathogenic bacterium Acinetobacter baumannii. The L. mucronata extract contained synergistic interactions (EFICI = 0.4181) and the S. terebinthifolia extract was non-interactive overall (EFICI = 0.9129). Quantifying interactions in the bioassay-guided fractionation of natural substances does not require additional experiments and can be useful to guide the experimental process and to support the development of standardized extracts as botanical drugs.

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MicroRNAs and long non-coding RNAs as potential candidates to target specific motifs of SARS-CoV-2

10.21203/rs.3.rs-80442/v1 ◽

2020 ◽

Author(s):

Lucia Natarelli ◽

Luca Parca ◽

Fabio Virgili ◽

Tommaso Mazza ◽

Christian Weber ◽

...

Keyword(s):

Experimental Studies ◽

Specific Treatment ◽

Leader Sequence ◽

The Novel ◽

Health Crisis ◽

Global Pandemic ◽

Pulmonary Arterial ◽

Non Coding Rnas ◽

Novel Coronavirus

Abstract The novel Coronavirus, SARS-CoV-2 disease (COVID-19) was defined as a global pandemic and induced a severe public health crisis in 2020. Covid-19 viral infection targets the human respiratory system and, at present, no specific treatment has been identified even though certain drugs have been studied and considered apparently effective in viral progression by reducing the complications in the lung epithelium. Researchers and clinicians are still struggling to find a vaccine or a specific innovative therapeutic strategy to counter COVID-19 infection.Here we describe our study indicating that SARS-CoV-2 genome contains motif sequences in the 5´UTR leader sequence that can be selectively recognized by specific human non-coding RNAs (ncRNAs), such as micro and long non-coding RNAs (miRNAs and lncRNA). Notably, some of these ncRNAs have been already utilized as oligo-based drugs in pulmonary and virus-associated diseases. We identified three selective motifs at the 5´UTR leader sequence of SARS-CoV-2 that allow viral recognition and binding of a specific group of miRNAs, some of them characterized by “GU” seed alignments. Additionally, one seed motif within miRNAs has been found to be able to bind the 5’UTR leader sequence. Among miRNAs having thermodynamically stable binding site against leader sequence and that are able interacted with Spike transcript some are involved in pulmonary arterial hypertension and anti-viral response, i.e. miR-204, miR-3661, and miR-1343. Moreover, several miRNA candidates have been already validated in vivo and specific oligo sequence are indeed available for their inhibition or overexpression.Four lncRNAs (H19, Hotair, Fendrr, and LINC05) directly interact with spike transcript (mRNA) and viral genome.In conclusion, we suggest that specific miRNAs and lncRNAs can be potential candidates to design oligonucleotide-drugs to treat COVID-19 and that our study can provide candidate hypothesis to be eventually tested in further experimental studies.

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A REVIEW ON THE ANTIPARASITIC DRUG IVERMECTIN FOR VARIOUS VIRAL INFECTIONS AND POSSIBILITIES OF USING IT FOR NOVEL SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2: NEW HOPE TO TREAT CORONAVIRUS DISEASE-2019

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2020.v13i8.38357 ◽

2020 ◽

pp. 21-27

Author(s):

TALHA JABEEN ◽

MOHD ABDUL KHADER ◽

SHAYESTHA JABEEN

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Viral Infections ◽

Sequence Similarity ◽

Drug Repurposing ◽

In Vivo Studies ◽

The Novel ◽

Antiparasitic Drug ◽

Coronavirus Infection ◽

Novel Coronavirus

The novel coronavirus infection has spread all over the world. With no specific drug or vaccine, the process of “drug repurposing” becomes a feasible solution. As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has 80% sequence similarity with the SARS-CoV, the nuclear import inhibitor “Ivermectin” (IVM) has recently been studied as a possible treatment option for coronavirus disease-2019 (COVID-19). The article aims to provide a review on structure and immunogenicity of SARS-CoV-2, indications of IVM for viral diseases, its possible mechanism on COVID-19 with a brief discussion on IVM structure, pharmacokinetics, adverse drug reactions, drug interactions, and contraindications. Further, we made possible comparisons of IVM with solidarity trial drugs and analyzed its major advantages, limitations and gave necessary recommendations for its use in future in vivo studies in the treatment of COVID-19.

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Rose Exctracts and In-Vitro Inhibition of SARS-CoV-2 Spike: The Ace-2 Interaction

Acta Medica Bulgarica ◽

10.2478/amb-2021-0022 ◽

2021 ◽

Vol 48 (2) ◽

pp. 41-44

Author(s):

M. Arokiaraj ◽

E. Menesson

Keyword(s):

Inhibitory Concentration ◽

Receptor Interaction ◽

The Novel ◽

Beneficial Effects ◽

Therapeutic Benefits ◽

Dependent Sequence ◽

The Rose ◽

Dose Dependent

Abstract Objective The study was performed to evaluate the novel potential of red rose extract to inhibit SARS-CoV-2 spike protein-Ace2 receptor interaction in vitro. Methods ACE2 receptors were His-labelled, and the interaction was studied by chemiluminescence after the addition of anti-His HRP and HRP substrate. The inhibition of SARS-CoV-2 and ACE2 was assessed in a dose-dependent sequence. Results The 50% inhibitory concentration was observed at 0.75 percent v/v of the rose extract, and the 90% inhibition was seen at about 1.8 percent v/v. Steam inhalation or nebulization could be simple methods of delivering rose extracts to the lower respiratory tract and pulmonary tissues. Conclusion Rose extracts have a potential for inhibition of SARS-CoV-2 and ACE2 receptor in vitro, which could add beneficial effects in Covid-19 treatment. Further tests need to be performed to study their therapeutic benefits in vivo.

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Specific Labeling of Protein Domains with Antibody Fragments

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100098587 ◽

1981 ◽

Vol 39 ◽

pp. 416-417

Author(s):

U. Aebi ◽

L.E. Buhle ◽

W.E. Fowler

Keyword(s):

Image Processing ◽

Specific Protein ◽

Antibody Fragments ◽

Supramolecular Organization ◽

Two Dimensional ◽

Ordered Structures ◽

Virus Capsids ◽

Processing Techniques

Many important supramolecular structures such as filaments, microtubules, virus capsids and certain membrane proteins and bacterial cell walls exist as ordered polymers or two-dimensional crystalline arrays in vivo. In several instances it has been possible to induce soluble proteins to form ordered polymers or two-dimensional crystalline arrays in vitro. In both cases a combination of electron microscopy of negatively stained specimens with analog or digital image processing techniques has proven extremely useful for elucidating the molecular and supramolecular organization of the constituent proteins. However from the reconstructed stain exclusion patterns it is often difficult to identify distinct stain excluding regions with specific protein subunits. To this end it has been demonstrated that in some cases this ambiguity can be resolved by a combination of stoichiometric labeling of the ordered structures with subunit-specific antibody fragments (e.g. Fab) and image processing of the electron micrographs recorded from labeled and unlabeled structures.

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Uptake, Internalization and Degradation of the Novel Plasminogen Activator Reteplase (BM 06.022) in the Rat

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649973 ◽

1995 ◽

Vol 74 (06) ◽

pp. 1501-1510 ◽

Cited By ~ 7

Author(s):

J Kuiper ◽

H van de Bilt ◽

U Martin ◽

Th J C van Berkel

Keyword(s):

Plasminogen Activator ◽

Liver Cells ◽

The Novel ◽

Uptake Mechanism ◽

Liver Uptake ◽

Lysosomal Compartment ◽

Β Phase ◽

Α Phase ◽

Parenchymal Liver

SummaryThe catabolism of the novel plasminogen activator reteplase (BM 06.022) was described. For this purpose BM 06.022 was radiolabelled with l25I or with the accumulating label l25I-tyramine cellobiose (l25I-TC).BM 06.022 was injected at a pharmacological dose of 380 μg/kg b.w. and it was cleared from the plasma in a biphasic manner with a half-life of about 1 min in the α-phase and t1/2of 20-28 min in the β-phase. 28% and 72% of the injected dose was cleared in the α-phase and β-phase, respectively. Initially liver, kidneys, skin, bones, lungs, spleen, and muscles contributed mainly to the plasma clearance. Only liver and the kidneys, however, were responsible for the uptake and subsequent degradation of BM 06.022 and contributed for 75% to the catabolism of BM 06.022. BM 06.022 was degraded in the lysosomal compartment of both organs. Parenchymal liver cells were responsible for 70% of the liver uptake of BM 06.022. BM 06.022 associated rapidly to isolated rat parenchymal liver cells and was subsequently degraded in the lysosomal compartment of these cells. BM 06.022 bound with low-affinity to the parenchymal liver cells (550 nM) and the binding of BM 06.022 could be displaced by t-PA (IC50 5.6 nM), indicating that the low-density lipoprotein receptor-related protein (LRP) could be involved in the binding of BM 06.022. GST-RAP, which is an inhibitor of LRP, could in vivo significantly inhibit the uptake of BM 06.022 in the liver.It is concluded that BM 06.022 is metabolized primarily in the liver and the kidneys. These organs take up and degrade BM 06.022 in the lysosomes. The uptake mechanism of BM 06.022 in the kidneys is unknown, while LRP is responsible for a low-affinity binding and uptake of BM 06.022 in parenchymal liver cells.

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Is the Rigidity of SARS-CoV-2 Spike Receptor-Binding Motif the Hallmark for Its Enhanced Infectivity and Pathogenesis?

10.26434/chemrxiv.12091260.v1 ◽

2020 ◽

Cited By ~ 2

Author(s):

Angelo Spinello ◽

Andrea Saltalamacchia ◽

Alessandra Magistrato

Keyword(s):

Health Economic ◽

Global Health ◽

Receptor Binding ◽

Binding Motif ◽

Spike Glycoprotein ◽

High Affinity ◽

Global Pandemic ◽

Medical Countermeasures ◽

High Infectivity ◽

Human Receptor

The latest outbreak of a new pathogenic coronavirus (SARS-CoV-2) is provoking a global health, economic and societal crisis. All-atom simulations enabled us to uncover the key molecular traits underlying the high affinity of SARS-CoV-2 spike glycoprotein towards its human receptor, providing a rationale to its high infectivity. Harnessing this knowledge can boost developing effective medical countermeasures to fight the current global pandemic.

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A Plug-and-Play Approach for the De Novo Generation of Dually Functionalised Bispecifics

10.26434/chemrxiv.8068184.v1 ◽

2019 ◽

Author(s):

Antoine Maruani ◽

Peter A. Szijj ◽

Calise Bahou ◽

João C. F. Nogueira ◽

Stephen Caddick ◽

...

Keyword(s):

De Novo ◽

Therapeutic Index ◽

Antibody Fragments ◽

Bispecific Antibodies ◽

Full Potential ◽

Mechanisms Of Resistance ◽

Large Excess ◽

Chemical Methods ◽

New Class ◽

Novel Approach

Diseases are multifactorial, with redundancies and synergies between various pathways. However, most of the antibody-based therapeutics in clinical trials and on the market interact with only one target thus limiting their efficacy. The targeting of multiple epitopes could improve the therapeutic index of treatment and counteract mechanisms of resistance. To this effect, a new class of therapeutics emerged: bispecific antibodies.Bispecific formation using chemical methods is rare and low yielding and/or requires a large excess of one of the two proteins to avoid homodimerisation. In order for chemically prepared bispecifics to deliver their full potential, high-yielding, modular and reliable cross-linking technologies are required. Herein, we describe a novel approach not only for the rapid and high-yielding chemical generation of bispecific antibodies from native antibody fragments, but also for the site-specific dual functionalisation of the resulting bioconjugates. Based on orthogonal clickable functional groups, this strategy enables the assembly of functionalised bispecifics with controlled loading in a modular and convergent manner.

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