ACE2-targeting monoclonal antibody as potent and broad-spectrum coronavirus blocker

AbstractThe evolution of coronaviruses, such as SARS-CoV-2, makes broad-spectrum coronavirus preventional or therapeutical strategies highly sought after. Here we report a human angiotensin-converting enzyme 2 (ACE2)-targeting monoclonal antibody, 3E8, blocked the S1-subunits and pseudo-typed virus constructs from multiple coronaviruses including SARS-CoV-2, SARS-CoV-2 mutant variants (SARS-CoV-2-D614G, B.1.1.7, B.1.351, B.1.617.1, and P.1), SARS-CoV and HCoV-NL63, without markedly affecting the physiological activities of ACE2 or causing severe toxicity in ACE2 “knock-in” mice. 3E8 also blocked live SARS-CoV-2 infection in vitro and in a prophylactic mouse model of COVID-19. Cryo-EM and “alanine walk” studies revealed the key binding residues on ACE2 interacting with the CDR3 domain of 3E8 heavy chain. Although full evaluation of safety in non-human primates is necessary before clinical development of 3E8, we provided a potentially potent and “broad-spectrum” management strategy against all coronaviruses that utilize ACE2 as entry receptors and disclosed an anti-coronavirus epitope on human ACE2.

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ACE2-Targeting Monoclonal Antibody As A “Pan” Coronavirus Blocker In Vitro and In A Mouse Model

10.1101/2020.11.11.375972 ◽

2020 ◽

Author(s):

Yuning Chen ◽

Yanan Zhang ◽

Renhong Yan ◽

Guifeng Wang ◽

Yuanyuan Zhang ◽

...

Keyword(s):

Monoclonal Antibody ◽

Mouse Model ◽

Management Strategy ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

Viral Epitope ◽

Neutralizing Monoclonal Antibody ◽

First Time ◽

Critical Site

AbstractCoronaviruses have caused three major outbreaks of infectious disease since the beginning of 21st century. Broad-spectrum strategies that can be utilized in both current and future coronavirus outbreaks and mutation-tolerant are sought after. Here we report a monoclonal antibody 3E8 targeting human angiotensin-converting enzyme 2 (ACE2) neutralized pseudo-typed coronaviruse SARS-CoV-2, SARS-CoV-2-D614G, SARS-CoV and HCoV-NL63, without affecting physiological activities of ACE2 or causing toxicity in mouse model. 3E8 also blocked live SARS-CoV-2 infection in vitro and in a mouse model of COVID-19. Cryo-EM studies revealed the binding site of 3E8 on ACE2 and identified Histone 34 of ACE2 as a critical site of anti-viral epitope. Overall, our work has provided a potential “pan” coronavirus management strategy and disclosed a “pan” anti-coronavirus epitope on human ACE2 for the first time.SummaryBlocking Multiple Coronaviruses by An ACE2-Neutralizing Monoclonal Antibody

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Virtual screening as therapeutic strategy of COVID-19 targeting angiotensin-converting enzyme 2

Frontiers in Molecular Immunology ◽

10.25082/fmi.2021.01.002 ◽

2021 ◽

Vol 2 (1) ◽

pp. 16-27

Author(s):

Zahra Sharifinia ◽

◽

Samira Asadi ◽

Mahyar Irani ◽

Abdollah Allahverdi ◽

...

Keyword(s):

Virtual Screening ◽

Angiotensin Converting Enzyme ◽

Host Cells ◽

Spike Protein ◽

Potential Drug ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

Approved Drugs ◽

Fda Approved Drugs

Objective: The receptor-binding domain (RBD) of the S1 domain of the SARS-CoV- 2 Spike protein performs a key role in the interaction with Angiotensin-converting enzyme 2 (ACE2), leading to both subsequent S2 domain-mediated membrane fusion and incorporation of viral RNA in host cells. Methods: In this study, we investigated the inhibitor’s targeted compounds through existing human ACE2 drugs to use as a future viral invasion. 54 FDA approved drugs were selected to assess their binding affinity to the ACE2 receptor. The structurebased methods via computational ones have been used for virtual screening of the best drugs from the drug database. Key Findings: The ligands “Cinacalcet” and “Levomefolic acid” highaffinity scores can be a potential drug preventing Spike protein of SARS-CoV-2 and human ACE2 interaction. Levomefolic acid from vitamin B family was proved to be a potential drug as a spike protein inhibitor in previous clinical and computational studies. Besides that, in this study, the capability of Levomefolic acid to avoid ACE2 and Spike protein of SARS-CoV-2 interaction is indicated. Therefore, it is worth to consider this drug for more in vitro investigations as ACE2 and Spike protein inhibition candidate. Conclusion: The two Cinacalcet and Levomefolic acid are the two ligands that have highest energy binding for human ACE2 blocking among 54 FDA approved drugs.

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DOCKING STUDY OF NOVEL N-SUBSTITUTED 2,5-BIS[(7-CHLOROQUINOLIN-4-YL)AMINO]PENTANOIC DERIVATIVES AS SELECTIVE HIGH-BINDER WITH ANGIOTENSIN CONVERTING ENZYME 2

INDIAN DRUGS ◽

10.53879/id.57.08.12425 ◽

2020 ◽

Vol 57 (08) ◽

pp. 16-24

Author(s):

Mohammed Oday Ezzat ◽

Basma M. Abd Razik ◽

Kutayba F. Dawood

Keyword(s):

Angiotensin Converting Enzyme ◽

Active Site ◽

Docking Study ◽

World Health ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

Pharmaceutical Properties ◽

Novel Coronavirus

The prevalence of a novel coronavirus (2019-nCoV) in the last few months represents a serious threat as a world health emergency concern. Angiotensin-converting enzyme 2 (ACE2) is the host cellular receptor for the respiratory syndrome of coronavirus epidemic in 2019 (2019-nCoV). In this work, the active site of ACE2 is successfully located by Sitmap prediction tool and validated by different marketed drugs. To design and discover new medical countermeasure drugs, we evaluate a total of 184 molecules of 7-chloro-N-methylquinolin-4-amine derivatives for binding affinity inside the crystal structure of ACE2 located active site. A novel series of N-substituted 2,5-bis[(7-chloroquinolin-4-yl)amino]pentanoic acid derivatives is generated and evaluated for a prospect as a lead compound for (2019-nCoV) medication with a docking score range of (-10.60 to -8.99) kcal/mol for the highest twenty derivatives. Moreover, the ADME pharmaceutical properties were evaluated for further proposed experimental evaluation in vitro or in vivo

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Uremic Apelin and Leucocytic Angiotensin-Converting Enzyme 2 in CKD Patients

Toxins ◽

10.3390/toxins12120742 ◽

2020 ◽

Vol 12 (12) ◽

pp. 742

Author(s):

Bogusz Trojanowicz ◽

Christof Ulrich ◽

Matthias Girndt

Keyword(s):

Angiotensin Converting Enzyme ◽

Healthy Controls ◽

Converting Enzyme ◽

Blood Pressure Lowering ◽

Angiotensin Converting Enzyme 2 ◽

Pressure Lowering ◽

The Impact ◽

Progression Of Atherosclerosis

Apelin peptides (APLN) serve as second substrates for angiotensin-converting enzyme 2 (ACE2) and, in contrast to angiotensin II (AngII), exert blood-pressure lowering and vasodilatation effects through binding to G-coupled APLN receptor (APLNR). ACE2-mediated cleavage of the APLN may reduce its vasodilatory effects, but decreased ACE2 may potentiate the hypotensive properties of APLN. The role of APLN in uremia is unclear. We investigated the correlations between serum-APLN, leucocytic APLNR, and ACE2 in 32 healthy controls (NP), 66 HD, and 24 CKD3–5 patients, and the impact of APLN peptides on monocytic behavior and ACE2 expression under uremic conditions in vitro. We observed that serum APLN and leucocytic APLNR or SLCO2B1 were significantly elevated in uremic patients and correlated with decreased ACE2 on uremic leucocytes. APLN-treated THP-1 monocytes revealed significantly increased APLNR and ACE2, and reduced TNFa, IL-6, and MCSF. Uremic toxins induced a dramatic increase of miR-421 followed by significant reduction of ACE2 transcripts, partially counteracted with APLN-13 and -36. APLN-36 triggered the most potent transmigration and reduction of endothelial adhesion. These results suggest that although APLN peptides may partly protect against the decay of monocytic ACE2 transcripts, uremic milieu is the most dominant modulator of local ACE2, and likely to contribute to the progression of atherosclerosis.

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Identification of prolyl carboxypeptidase as an alternative enzyme for processing of renal angiotensin II using mass spectrometry

AJP Cell Physiology ◽

10.1152/ajpcell.00346.2012 ◽

2013 ◽

Vol 304 (10) ◽

pp. C945-C953 ◽

Cited By ~ 28

Author(s):

Nadja Grobe ◽

Nathan M. Weir ◽

Orly Leiva ◽

Frank S. Ong ◽

Kenneth E. Bernstein ◽

...

Keyword(s):

Ph Control ◽

Mass Spectrometric ◽

Acidic Ph ◽

Ang Ii ◽

Converting Enzyme ◽

Wild Type ◽

Angiotensin Converting Enzyme 2 ◽

Genetic Mouse Models

Angiotensin-converting enzyme 2 (ACE2) catalyzes conversion of ANG II to ANG-(1–7). The present study uses newly established proteomic approaches and genetic mouse models to examine the contribution of alternative renal peptidases to ACE2-independent formation of ANG-(1–7). In situ and in vitro mass spectrometric characterization showed that substrate concentration and pH control renal ANG II processing. At pH ≥6, ANG-(1–7) formation was significantly reduced in ACE2 knockout (KO) mice. However, at pH <6, formation of ANG-(1–7) in ACE2 KO mice was similar to that in wild-type (WT) mice, suggesting alternative peptidases for renal ANG II processing. Furthermore, the dual prolyl carboxypeptidase (PCP)-prolyl endopeptidase (PEP) inhibitor Z-prolyl-prolinal reduced ANG-(1–7) formation in ACE2 KO mice, while the ACE2 inhibitor MLN-4760 had no effect. Unlike the ACE2 KO mice, ANG-(1–7) formation from ANG II in PEP KO mice was not different from that in WT mice at any tested pH. However, at pH 5, this reaction was significantly reduced in kidneys and urine of PCP-depleted mice. In conclusion, results suggest that ACE2 metabolizes ANG II in the kidney at neutral and basic pH, while PCP catalyzes the same reaction at acidic pH. This is the first report demonstrating that renal ANG-(1–7) formation from ANG II is independent of ACE2. Elucidation of ACE2-independent ANG-(1–7) production pathways may have clinically important implications in patients with metabolic and renal disease.

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Uncontrolled mass use of hydroxychloroquine

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11ispl1.2734 ◽

2020 ◽

Vol 11 (SPL1) ◽

pp. 396-398

Author(s):

Tugolbai Tagaev ◽

Sagynali Mamatov ◽

Farida Imanalieva ◽

Vityala Yethindra ◽

Altynai Zhumabekova ◽

...

Keyword(s):

Clinical Trials ◽

Therapeutic Drug Monitoring ◽

Severe Acute Respiratory Syndrome ◽

Adverse Effects ◽

Medical Advice ◽

Therapeutic Drug ◽

Converting Enzyme ◽

Weak Base ◽

Angiotensin Converting Enzyme 2

Hydroxychloroquine (HCQ) has previously been shown to inhibit coronavirus replication in vitro. But antiviral properties mechanisms are not well known, HCQ is a weak base that accumulates in lysosomes, modifies their pH, and interferes with some enzymes. In the lack of confirmed efficacy, the initial potential risk is not to expose patients to adverse effects. However, results from preliminary clinical studies have drawn inconclusive results regarding the efficacy of HCQ in coronavirus disease 2019 (COVID-19), due to several important weaknesses in research methodologies. Hypokalemia often occurs in patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), possibly due to the particular tropism of SARS-CoV-2 with regard to Angiotensin-converting enzyme 2 (ACE2). The wide use of HCQ, even against medical advice, will show an impact on ongoing clinical trials. It is important that we can recruite COVID-19 patients in these research studies to generate appropriate data regarding drugs that show promising efficacy against COVID-19. Currently, only doctors should be allowed to prescribe HCQ, and treatment should be confined to hospital settings, with proper cardiac and therapeutic drug monitoring.

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A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent in vitro

10.1101/2020.09.18.301952 ◽

2020 ◽

Cited By ~ 1

Author(s):

Tianshu Xiao ◽

Jianming Lu ◽

Jun Zhang ◽

Rebecca I. Johnson ◽

Lindsay G.A. McKay ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Renin Angiotensin System ◽

Negative Regulator ◽

Peptidase Activity ◽

Converting Enzyme ◽

Angiotensin Ii Receptor ◽

Angiotensin System ◽

Angiotensin Converting Enzyme 2

AbstractEffective intervention strategies are urgently needed to control the COVID-19 pandemic. Human angiotensin-converting enzyme 2 (ACE2) is a carboxypeptidase that forms a dimer and serves as the cellular receptor for SARS-CoV-2. It is also a key negative regulator of the renin-angiotensin system (RAS), conserved in mammals, which modulates vascular functions. We report here the properties of a trimeric ACE2 variant, created by a structure-based approach, with binding affinity of ~60 pM for the spike (S) protein of SARS-CoV-2, while preserving the wildtype peptidase activity as well as the ability to block activation of angiotensin II receptor type 1 in the RAS. Moreover, the engineered ACE2 potently inhibits infection of SARS-CoV-2 in cell culture. These results suggest that engineered, trimeric ACE2 may be a promising anti-SARS-CoV-2 agent for treating COVID-19.

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Stapled peptides based on Human Angiotensin-Converting Enzyme 2 (ACE2) potently inhibit SARS-CoV-2 infection in vitro

10.1101/2020.08.25.266437 ◽

2020 ◽

Author(s):

Francesca Curreli ◽

Sofia M B Victor ◽

Shahad Ahmed ◽

Aleksandra Drelich ◽

Xiaohe Tong ◽

...

Keyword(s):

Antiviral Activity ◽

Angiotensin Converting Enzyme ◽

Host Cells ◽

Converting Enzyme ◽

Stapled Peptides ◽

Angiotensin Converting Enzyme 2 ◽

Stapled Peptide ◽

Human Lung Cells ◽

Critical Binding

AbstractSARS-CoV-2 uses human angiotensin-converting enzyme 2 (ACE2) as the primary receptor to enter host cells and initiate the infection. The critical binding region of ACE2 is a ∼30 aa long helix. Here we report the design of four stapled peptides based on the ACE2 helix, which is expected to bind to SARS-CoV-2 and prevent the binding of the virus to the ACE2 receptor and disrupt the infection. All stapled peptides showed high helical contents (50-94% helicity). On the contrary, the linear control peptide NYBSP-C showed no helicity (19%). We have evaluated the peptides in a pseudovirus based single-cycle assay in HT1080/ACE2 and human lung cells A549/ACE2, overexpressing ACE2. Three of the four stapled peptides showed potent antiviral activity in HT1080/ACE2 (IC50: 1.9 – 4.1 µM) and A549/ACE2 cells (IC50: 2.2 – 2.8 µM). The linear peptides NYBSP-C and the double-stapled peptide StRIP16, used as controls, showed no antiviral activity. Most significantly, none of the stapled peptides show any cytotoxicity at the highest dose tested. We also evaluated the antiviral activity of the peptides by infecting Vero E6 cells with the replication-competent authentic SARS-CoV-2 (US_WA-1/2020). NYBSP-1 was the most efficient preventing the complete formation of cytopathic effects (CPEs) at an IC100 17.2 µM. NYBSP-2 and NYBSP-4 also prevented the formation of the virus-induced CPE with an IC100 of about 33 µM. We determined the proteolytic stability of one of the most active stapled peptides, NYBSP-4, in human plasma, which showed a half-life (T1/2) of >289 min.

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Preclinical Evaluation and Pilot Clinical Study of Al18F-DX600-BCH for non-invasive PET Mapping of Angiotensin Converting Enzyme 2 in Mammal

10.21203/rs.3.rs-1059130/v1 ◽

2021 ◽

Author(s):

Jin Ding ◽

Qian Zhang ◽

Jinquan Jiang ◽

Nina Zhou ◽

Zilei Wang ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Pet Imaging ◽

Transmembrane Protein ◽

Radiochemical Yield ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

Non Invasive ◽

Pet Ct ◽

Pilot Clinical Study

Abstract Angiotensin-converting enzyme 2 (ACE2), a transmembrane protein, is the main entry point for certain coronaviruses including the new coronavirus SARS-CoV-2 to enter cells. Synthesizing the PET imaging probe Al18F-DX600-BCH which is high-affinity ACE2 is aim to detect the expression of ACE2 in body and monitor the therapeutic effect. The Al18F-DX600-BCH was obtained manually with a 20.4% ± 5.2% radiochemical yield without attenuation correction and an over 99% purified radiochemical purity, being stable in vitro within 4 hours and cleared rapidly in blood (the half-lives of the distribution phase and clearance phase were 2.12 min and 25.31 min, respectively). Results of both biodistribution and PET imaging showed that Al18F-DX600-BCH was highly accumulated in the kidney (SUVkidney/normal > 50), and specific uptake in testis (SUVtestis/normal > 10) was observed in rat images. The kidney (++), gastrointestinal (++) and bronchial (+++) cells were evidenced of ACE2 positive by IHC staining of rats. A total of 10 volunteers were enrolled and received PET/CT 1 hour and 2 hours after injection or dynamic PET/CT during 0-330 seconds (NCT04542863), from which strong radioactivity accumulation was mostly observed in the genitourinary system (SUVrenal cortex = 32.00, SUVtestis = 4.56), and moderate accumulation in conjunctiva and nasal mucosa for several cases. This work firstly reported the probe Al18F-DX600-BCH targeting ACE2, conducting preliminary preclinical experiments and a total of 10 clinical transformations, which demonstrated the potential and possibility of non-invasive mapping of ACE2. Trial registration: ClinicalTrials.gov NCT04542863. Registered 9 September 2020.

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Dalbavancin binds ACE2 to block its interaction with SARS-CoV-2 spike protein and is effective in inhibiting SARS-CoV-2 infection in animal models

Cell Research ◽

10.1038/s41422-020-00450-0 ◽

2020 ◽

Vol 31 (1) ◽

pp. 17-24 ◽

Cited By ~ 1

Author(s):

Gan Wang ◽

Meng-Li Yang ◽

Zi-Lei Duan ◽

Feng-Liang Liu ◽

Lin Jin ◽

...

Keyword(s):

Clinical Trials ◽

Spike Protein ◽

Drug Candidate ◽

Peptide Drug ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

High Affinity ◽

Plasma Half Life

AbstractInfection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic worldwide. Currently, however, no effective drug or vaccine is available to treat or prevent the resulting coronavirus disease 2019 (COVID-19). Here, we report our discovery of a promising anti-COVID-19 drug candidate, the lipoglycopeptide antibiotic dalbavancin, based on virtual screening of the FDA-approved peptide drug library combined with in vitro and in vivo functional antiviral assays. Our results showed that dalbavancin directly binds to human angiotensin-converting enzyme 2 (ACE2) with high affinity, thereby blocking its interaction with the SARS-CoV-2 spike protein. Furthermore, dalbavancin effectively prevents SARS-CoV-2 replication in Vero E6 cells with an EC50 of ~12 nM. In both mouse and rhesus macaque models, viral replication and histopathological injuries caused by SARS-CoV-2 infection are significantly inhibited by dalbavancin administration. Given its high safety and long plasma half-life (8–10 days) shown in previous clinical trials, our data indicate that dalbavancin is a promising anti-COVID-19 drug candidate.

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