Potent in vitro anti-SARS-CoV-2 activity by gallinamide A and analogues via inhibition of cathepsin L

2020 ◽  
Author(s):  
Anneliese S. Ashhurst ◽  
Arthur H. Tang ◽  
Pavla Fajtová ◽  
Michael Yoon ◽  
Anupriya Aggarwal ◽  
...  
Keyword(s):  
Drug Target ◽  
Cathepsin L ◽  
Antiviral Drug ◽  
Cell Entry ◽  
Marine Natural Product ◽  
Viral Proteases ◽  
Cathepsin Proteases ◽  
Human Cathepsin ◽  
Nanomolar Range

AbstractThe emergence of SARS-CoV-2 in late 2019, and the subsequent COVID-19 pandemic, has led to substantial mortality, together with mass global disruption. There is an urgent need for novel antiviral drugs for therapeutic or prophylactic application. Cathepsin L is a key host cysteine protease utilized by coronaviruses for cell entry and is recognized as a promising drug target. The marine natural product, gallinamide A and several synthetic analogues, were identified as potent inhibitors of cathepsin L activity with IC50 values in the picomolar range. Lead molecules possessed selectivity over cathepsin B and other related human cathepsin proteases and did not exhibit inhibitory activity against viral proteases Mpro and PLpro. We demonstrate that gallinamide A and two lead analogues potently inhibit SARS-CoV-2 infection in vitro, with EC50 values in the nanomolar range, thus further highlighting the potential of cathepsin L as a COVID-19 antiviral drug target.

scholarly journals Structure and inhibition of the SARS-CoV-2 main protease reveals strategy for developing dual inhibitors against Mpro and cathepsin L

2020 ◽  
Author(s):  
Michael Dominic Sacco ◽  
Chunlong Ma ◽  
Panagiotis Lagarias ◽  
Ang Gao ◽  
Julia Alma Townsend ◽  
...  
Keyword(s):  
Viral Entry ◽  
Cathepsin L ◽  
Antiviral Drug ◽  
Binding Mode ◽  
Calpain Inhibitor ◽  
Catalytic Core ◽  
Main Protease ◽  
Dual Inhibitors ◽  
Calpain Inhibitors

AbstractThe main protease (Mpro) of SARS-CoV-2, the pathogen responsible for the COVID-19 pandemic, is a key antiviral drug target. While most SARS-CoV-2 Mpro inhibitors have a γ-lactam glutamine surrogate at the P1 position, we recently discovered several Mpro inhibitors have hydrophobic moieties at the P1 site, including calpain inhibitors II/XII, which are also active against human cathepsin L, a host-protease that is important for viral entry. To determine the binding mode of these calpain inhibitors and establish a structure-activity relationship, we solved X-ray crystal structures of Mpro in complex with calpain inhibitors II and XII, and three analogues of GC-376, one of the most potent Mpro inhibitors in vitro. The structure of Mpro with calpain inhibitor II confirmed the S1 pocket of Mpro can accommodate a hydrophobic methionine side chain, challenging the idea that a hydrophilic residue is necessary at this position. Interestingly, the structure of calpain inhibitor XII revealed an unexpected, inverted binding pose where the P1’ pyridine inserts in the S1 pocket and the P1 norvaline is positioned in the S1’ pocket. The overall conformation is semi-helical, wrapping around the catalytic core, in contrast to the extended conformation of other peptidomimetic inhibitors. Additionally, the structures of three GC-376 analogues UAWJ246, UAWJ247, and UAWJ248 provide insight to the sidechain preference of the S1’, S2, S3 and S4 pockets, and the superior cell-based activity of the aldehyde warhead compared with the α-ketoamide. Taken together, the biochemical, computational, structural, and cellular data presented herein provide new directions for the development of Mpro inhibitors as SARS-CoV-2 antivirals.

Post-transcriptional regulation of human cathepsin L expression

2011 ◽  
Vol 392 (5) ◽  
Author(s):  
Shivani Mittal ◽  
Riyaz A. Mir ◽  
Shyam S. Chauhan
Keyword(s):  
Splice Variants ◽  
Cathepsin L ◽  
Live Cells ◽  
Luciferase Reporter ◽  
Entry Site ◽  
Lower Efficiency ◽  
Internal Ribosome Entry ◽  
Lysosomal Protease ◽  
Human Cathepsin

AbstractThe expression of cathepsin L, a lysosomal protease, is known to be elevated in cancer and other pathologies. Multiple splice variants of human cathepsin L with variable 5′UTRs exist, which encode for the same protein. Previously we have observed that variant hCATL A (bearing the longest 5′UTR) was translatedin vitrowith significantly lower efficiency than variant hCATL AIII (bearing the shortest 5′UTR). Contrary to these findings, results of the present study reveal that in cancer cells, hCATL A mRNA exhibits higher translatability in spite of having lower stability than AIII. This is the first report demonstrating a highly contrasting trend in translation efficiencies of hCATL variants in rabbit reticulocytes and live cells. Expression from chimeric mRNAs containing 5′UTRs of A or AIII upstream to luciferase reporter cDNA established the A UTR to be the sole determinant for this effect. Transient transfections of bicistronic plasmids and mRNAs confirmed the presence of a functional Internal Ribosome Entry Site in this UTR. Our data suggest that differential stability and translation initiation modes mediated by the 5′UTRs of human cathepsin L variants are involved in regulating its expression.

Splice Variants of Human Cathepsin L mRNA Show Different Expression Rates

2001 ◽  
Vol 382 (11) ◽  
pp. 1583-1591 ◽  
Author(s):  
Abulizi Abudula ◽  
Winfried Rommerskirch ◽  
Ekkehard Weber ◽  
Dagmar Günther ◽  
Bernd Wiederanders
Keyword(s):  
Malignant Tumors ◽  
Splice Variants ◽  
Cathepsin L ◽  
Mrna Levels ◽  
Kidney Tumors ◽  
Exon 1 ◽  
Expression Rate ◽  
Human Cathepsin

Abstract Human cathepsin L (hCATL) mRNA occurs in vivo in at least three splice variants. They differ in the length of exon 1, which comprises 278 nucleotides (hCATLA), 188 nucleotides (hCATLA2) and 132 nucleotides (hCATLA3), respectively. We describe here the shortest variant for the first time. This form is predominant in all tissues and cells examined so far, including malignant tumors. We studied the expression rate of the three mRNA variants in order to explain why malignant kidney tumors show low cathepsin L activity despite of high mRNA levels. The variant hCATLA3 showed the highest expression rate in vitro and in vivo. Based on these results, we suggest a cisacting element on human cathepsin L mRNA which can be bound by a negative transacting regulator, thus leading to reduced expression rates.

scholarly journals Understanding the phase separation characteristics of nucleocapsid protein provides a new therapeutic opportunity against SARS-CoV-2

2020 ◽  
Author(s):  
Dan Zhao ◽  
Weifan Xu ◽  
Xiaofan Zhang ◽  
Xiaoting Wang ◽  
Enming Yuan ◽  
...  
Keyword(s):  
Phase Separation ◽  
Drug Target ◽  
Antiviral Drug ◽  
Viral Rna ◽  
N Protein ◽  
Therapeutic Opportunity ◽  
Further Development ◽  
Liquid Liquid Phase Separation

AbstractThe ongoing coronavirus disease 2019 (COVID-19) pandemic has raised an urgent need to develop effective therapeutics against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As a potential antiviral drug target, the nucleocapsid (N) protein of SARS-CoV-2 functions as a viral RNA chaperone and plays vital and multifunctional roles during the life cycle of coronavirus1-3. In this study, we discovered that the N protein of SARS-CoV-2 undergoes liquid-liquid phase separation (LLPS) both in vitro and in vivo, which is further modulated by viral RNA. In addition, we found that, the core component of the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2, nsp12, preferentially partitions into the N protein condensates. Moreover, we revealed that, two small molecules, i.e., CVL218 and PJ34, can be used to intervene the N protein driven phase separation and loosen the compact structures of the condensates of the N-RNA-nsp12 complex of SARS-CoV-2. The discovery of the LLPS-mediated interplay between N protein and nsp12 and the corresponding modulating compounds illuminates a feasible way to improve the accessibility of antiviral drugs (e.g., remdesivir) to their targets (e.g., nsp12/RdRp), and thus may provide useful hints for further development of effective therapeutic strategies against SARS-CoV-2.

scholarly journals Cathepsin L Functionally Cleaves the Severe Acute Respiratory Syndrome Coronavirus Class I Fusion Protein Upstream of Rather than Adjacent to the Fusion Peptide

2008 ◽  
Vol 82 (17) ◽  
pp. 8887-8890 ◽  
Author(s):  
Berend Jan Bosch ◽  
Willem Bartelink ◽  
Peter J. M. Rottier
Keyword(s):  
Fusion Proteins ◽  
Cathepsin L ◽  
Fusion Peptide ◽  
Cell Entry ◽  
Spike Protein ◽  
Class I ◽  
Viral Fusion ◽  
Cathepsin Proteases ◽  
Viral Fusion Proteins ◽  
Binding Subunit

ABSTRACT Unlike other class I viral fusion proteins, spike proteins on severe acute respiratory sydrome coronavirus virions are uncleaved. As we and others have demonstrated, infection by this virus depends on cathepsin proteases present in endosomal compartments of the target cell, suggesting that the spike protein acquires its fusion competence by cleavage during cell entry rather than during virion biogenesis. Here we demonstrate that cathepsin L indeed activates the membrane fusion function of the spike protein. Moreover, cleavage was mapped to the same region where, in coronaviruses carrying furin-activated spikes, the receptor binding subunit of the protein is separated from the membrane-anchored fusion subunit.

scholarly journals Antiplasmodial Activity and Mechanism of Action of RSM-932A, a Promising Synergistic Inhibitor of Plasmodium falciparum Choline Kinase

2013 ◽  
Vol 57 (12) ◽  
pp. 5878-5888 ◽  
Author(s):  
Tahl Zimmerman ◽  
Carlos Moneriz ◽  
Amalia Diez ◽  
José Manuel Bautista ◽  
Teresa Gómez del Pulgar ◽  
...  
Keyword(s):  
Mechanism Of Action ◽  
Drug Target ◽  
Maturation Stage ◽  
Choline Kinase ◽  
Content Type ◽  
Mechanism Of Inhibition ◽  
Resistant Strains ◽  
Parasite Egress ◽  
Nanomolar Range

ABSTRACTWe have investigated the mechanism of action of inhibition of the choline kinase ofP. falciparum(p.f.-ChoK) by two inhibitors of the human ChoKα, MN58b and RSM-932A, which have previously been shown to be potent antitumoral agents. The efficacy of these inhibitors againstp.f.-ChoK is investigated using enzymatic andin vitroassays. While MN58b may enter the choline/phosphocholine binding site, RSM-932A appears to have an altogether novel mechanism of inhibition and is synergistic with respect to both choline and ATP. A model of inhibition for RSM-932A in which this inhibitor trapsp.f.-ChoK in a phosphorylated intermediate state blocking phosphate transfer to choline is presented. Importantly, MN58b and RSM-932A havein vitroinhibitory activity in the low nanomolar range and are equally effective against chloroquine-sensitive and chloroquine-resistant strains. RSM-932A and MN58b significantly reduced parasitemia and induced the accumulation of trophozoites and schizonts, blocking intraerythrocytic development and interfering with parasite egress or invasion, suggesting a delay of the parasite maturation stage. The present data provide two new potent structures for the development of antimalarial compounds and validatep.f.-ChoK as an accessible drug target against the parasite.

scholarly journals Novel Dihydroorotate Dehydrogenase Inhibitors with Potent Interferon-Independent Antiviral Activity against Mammarenaviruses In Vitro

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 821 ◽  
Author(s):  
Yu-Jin Kim ◽  
Beatrice Cubitt ◽  
Yingyun Cai ◽  
Jens H. Kuhn ◽  
Daniel Vitt ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
De Novo ◽  
Lymphocytic Choriomeningitis ◽  
Human Infection ◽  
Cell Entry ◽  
Dihydroorotate Dehydrogenase ◽  
Chronic Infections ◽  
Natural Hosts ◽  
Nanomolar Range

Mammarenaviruses cause chronic infections in rodents, which are their predominant natural hosts. Human infection with some of these viruses causes high-consequence disease, posing significant issues in public health. Currently, no FDA-licensed mammarenavirus vaccines are available, and anti-mammarenavirus drugs are limited to an off-label use of ribavirin, which is only partially efficacious and associated with severe side effects. Dihydroorotate dehydrogenase (DHODH) inhibitors, which block de novo pyrimidine biosynthesis, have antiviral activity against viruses from different families, including Arenaviridae, the taxonomic home of mammarenaviruses. Here, we evaluate five novel DHODH inhibitors for their antiviral activity against mammarenaviruses. All tested DHODH inhibitors were potently active against lymphocytic choriomeningitis virus (LCMV) (half-maximal effective concentrations [EC50] in the low nanomolar range, selectivity index [SI] > 1000). The tested DHODH inhibitors did not affect virion cell entry or budding, but rather interfered with viral RNA synthesis. This interference resulted in a potent interferon-independent inhibition of mammarenavirus multiplication in vitro, including the highly virulent Lassa and Junín viruses.

scholarly journals Antiviral efficacy in vivo of the anti-human immunodeficiency virus bicyclam SDZ SID 791 (JM 3100), an inhibitor of infectious cell entry.

1996 ◽  
Vol 40 (3) ◽  
pp. 750-754 ◽  
Author(s):  
R Datema ◽  
L Rabin ◽  
M Hincenbergs ◽  
M B Moreno ◽  
S Warren ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Antiviral Agents ◽  
Antiviral Drug ◽  
Virus Production ◽  
Significant Inhibition ◽  
Cell Entry ◽  
Immunodeficiency Virus ◽  
Antiviral Chemotherapy

SID 791, a bicyclam inhibiting human immunodeficiency virus (HIV) replication in vitro by blocking virus entry into cells, is an effective inhibitor of virus production and of depletion of human CD4+ T cells in HIV type 1-infected SCID-hu Thy/Liv mice. Steady levels of 100 ng of SID 791 or higher per ml in plasma resulted in statistically significant inhibition of p24 antigen formation. Daily injections of SID 791 caused a dose-dependent decrease in viremia, and this inhibition could be potentiated by coadministration of zidovudine or didanose. The present study suggests that SID 791 alone or in combination with licensed antiviral agents may decrease the virus load in HIV-infected patients and, by extension, that the infectious cell entry step is a valid target for antiviral chemotherapy of HIV disease. The SCID-hu Thy/Liv model in effect provides a rapid means of assessing the potential of compounds with novel modes of antiviral action, as well as the potential of antiviral drug combinations.

Biphasic modulation of human cathepsin L by a novel cyanobacterial depsipeptide

Planta Medica ◽  
2015 ◽  
Vol 81 (11) ◽  
Author(s):  
B Miller ◽  
M Bertin ◽  
V Hook ◽  
WH Gerwick
Keyword(s):  
Cathepsin L ◽  
Human Cathepsin
Sign in / Sign up

Export Citation Format

Share Document