scholarly journals Combined computational and cellular screening identifies synergistic inhibition of SARS-CoV-2 by lenvatinib and remdesivir

2021 ◽  
Author(s):  
Marie O. Pohl ◽  
Idoia Busnadiego ◽  
Francesco Marrafino ◽  
Lars Wiedmer ◽  
Annika Hunziker ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Therapeutic Option ◽  
Protease Inhibition ◽  
Antiviral Compounds ◽  
Main Protease ◽  
Ic50 Value ◽  
Anti Cancer ◽  
Synergistic Mechanism ◽  
Host Cell Entry

Rapid repurposing of existing drugs as new therapeutics for COVID-19 has been an important strategy in the management of disease severity during the ongoing SARS-CoV-2 pandemic. Here, we screened by high-throughput docking 6,000 compounds within the DrugBank library for their potential to bind and inhibit the SARS-CoV-2 3CL main protease, a chymotrypsin-like enzyme that is essential for viral replication. For 19 candidate hits, parallel in vitro fluorescence-based protease-inhibition assays and Vero-CCL81 cell-based SARS-CoV-2 replication-inhibition assays were performed. One hit, diclazuril (an investigational anti-protozoal compound), was validated as a SARS-CoV-2 3CL main protease inhibitor in vitro (IC50 value of 29 μM) and modestly inhibited SARS-CoV-2 replication in Vero-CCL81 cells. Another hit, lenvatinib (approved for use in humans as an anti-cancer treatment), could not be validated as a SARS-CoV-2 3CL main protease inhibitor in vitro, but serendipitously exhibited a striking functional synergy with the approved nucleoside analogue remdesivir to inhibit SARS-CoV-2 replication in Vero-CCL81 cells. Lenvatinib is a broadly-acting host receptor tyrosine kinase (RTK) inhibitor, but the synergistic effect with remdesivir was not observed with other approved RTK inhibitors (such as pazopanib or sunitinib), suggesting that the mechanism-of-action is independent of host RTKs. Furthermore, time-of-addition studies revealed that lenvatinib/remdesivir synergy probably targets SARS-CoV-2 replication subsequent to host-cell entry. Our study shows that combining computational and cellular screening is an efficient means to identify existing drugs with repurposing potential as antiviral compounds. Future studies should aim at understanding and optimizing the lenvatinib/remdesivir synergistic mechanism as a therapeutic option.

scholarly journals Combined computational and cellular screening identifies synergistic inhibition of SARS-CoV-2 by lenvatinib and remdesivir

2021 ◽  
Vol 102 (7) ◽  
Author(s):  
Marie O. Pohl ◽  
Idoia Busnadiego ◽  
Francesco Marrafino ◽  
Lars Wiedmer ◽  
Annika Hunziker ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Therapeutic Option ◽  
Cell Entry ◽  
Protease Inhibition ◽  
Antiviral Compounds ◽  
Main Protease ◽  
Anti Cancer ◽  
Synergistic Mechanism ◽  
Host Cell Entry

Rapid repurposing of existing drugs as new therapeutics for COVID-19 has been an important strategy in the management of disease severity during the ongoing SARS-CoV-2 pandemic. Here, we used high-throughput docking to screen 6000 compounds within the DrugBank library for their potential to bind and inhibit the SARS-CoV-2 3 CL main protease, a chymotrypsin-like enzyme that is essential for viral replication. For 19 candidate hits, parallel in vitro fluorescence-based protease-inhibition assays and Vero-CCL81 cell-based SARS-CoV-2 replication-inhibition assays were performed. One hit, diclazuril (an investigational anti-protozoal compound), was validated as a SARS-CoV-2 3 CL main protease inhibitor in vitro (IC50 value of 29 µM) and modestly inhibited SARS-CoV-2 replication in Vero-CCL81 cells. Another hit, lenvatinib (approved for use in humans as an anti-cancer treatment), could not be validated as a SARS-CoV-2 3 CL main protease inhibitor in vitro, but serendipitously exhibited a striking functional synergy with the approved nucleoside analogue remdesivir to inhibit SARS-CoV-2 replication, albeit this was specific to Vero-CCL81 cells. Lenvatinib is a broadly-acting host receptor tyrosine kinase (RTK) inhibitor, but the synergistic effect with remdesivir was not observed with other approved RTK inhibitors (such as pazopanib or sunitinib), suggesting that the mechanism-of-action is independent of host RTKs. Furthermore, time-of-addition studies revealed that lenvatinib/remdesivir synergy probably targets SARS-CoV-2 replication subsequent to host-cell entry. Our work shows that combining computational and cellular screening is a means to identify existing drugs with repurposing potential as antiviral compounds. Future studies could be aimed at understanding and optimizing the lenvatinib/remdesivir synergistic mechanism as a therapeutic option.

scholarly journals Kunitz type protease inhibitor EgKI-1 from the canine tapeworm Echinococcus granulosus as a promising anti-cancer therapeutic

2018 ◽  
Author(s):  
Shiwanthi L Ranasinghe ◽  
Glen M Boyle ◽  
Katja Fischer ◽  
Jeremy Potriquet ◽  
Jason P Mulvenna ◽  
...  
Keyword(s):  
Cell Growth ◽  
Protease Inhibitor ◽  
Hydatid Cyst ◽  
Echinococcus Granulosus ◽  
Cancer Cell ◽  
Dependent Manner ◽  
Anti Cancer ◽  
Kunitz Type ◽  
Kunitz Type Protease Inhibitor

AbstractEgKI-1, a member of the Kunitz type protease inhibitor family, is highly expressed by the oncosphere of the canine tapeworm Echinococcus granulosus, the stage that is infectious to humans and ungulates, giving rise to a hydatid cyst localized to the liver and other organs. Larval protoscoleces, which develop within the hydatid cyst, have been shown to possess anti-cancer properties, although the precise molecules involved have not been identified. We show that recombinant EgKI-1 inhibits the growth and migration of a range of human cancers including breast, melanoma and cervical cancer cell lines in a dose-dependent manner in vitro without affecting normal cell growth. Furthermore, EgKI-1 treatment arrested the cancer cell growth by disrupting the cell cycle and induced apoptosis of cancer cells in vitro. An in vivo model of triple negative breast cancer (MDA-MB-231) in BALB/c nude mice showed significant tumor growth reduction in EgKI-1-treated mice compared with controls. These findings indicate that EgKI-1 shows promise for future development as an anti-cancer therapeutic.

scholarly journals Favipiravir May Acts as COVID-19 Main Protease PDB ID 6LU7 Inhibitor: Docking Analysis

2020 ◽  
Vol 10 (6) ◽  
pp. 6821-6828 ◽  
Keyword(s):  
Amino Acids ◽  
Protease Inhibitor ◽  
Essential Amino Acids ◽  
Antiviral Drugs ◽  
Binding Pattern ◽  
Protease Inhibition ◽  
Docking Analysis ◽  
Protease Enzyme ◽  
Main Protease ◽  
Polymerase Inhibitor

Coronavirus is a well-known threat to the human being in the form of COVID-19. Virus replication may be controlled by inhibition of protease enzyme. Hence, well known 13 antiviral drugs have been observed by docking analysis for understanding the binding pattern of drugs with COVID-19 main protease PDB ID: 6LU7 for any possibilities of protease inhibition. For docking analysis PyRx- Python Prescription 0.8 was used. This analysis reveals that the essential amino acids involved in binding of antiviral drugs to COVID-19 main protease PDB ID: 6LU7 are Glycine (Gly), Serine (Ser), Cysteine (Cys), Leucine (Leu), Asparagine (Asn), Glutamine (Gln), Glutamic acid (Glu) and Threonine (Thr). After docking analysis, it was observed that Favipiravir maybe act as COVID-19 main protease inhibitor despite being vRNA polymerase inhibitor and may further be used in the treatment of COVID-19 infection.

scholarly journals Suppressors of a Host Range Mutation in the Rabbitpox Virus Serpin SPI-1 Map to Proteins Essential for Viral DNA Replication

2005 ◽  
Vol 79 (14) ◽  
pp. 9168-9179 ◽  
Author(s):  
Benjamin G. Luttge ◽  
Richard W. Moyer
Keyword(s):  
Dna Replication ◽  
Protease Inhibitor ◽  
Host Range ◽  
Suppressor Mutation ◽  
Protease Inhibition ◽  
Viral Dna ◽  
Viral Dna Replication ◽  
Range Function ◽  
Rabbitpox Virus

ABSTRACT The orthopoxvirus serpin SPI-1 is an intracellular serine protease inhibitor that is active against cathepsin G in vitro. Rabbitpox virus (RPV) mutants with deletions of the SPI-1 gene grow on monkey kidney cells (CV-1) but do not plaque on normally permissive human lung carcinoma cells (A549). This reduced-host-range (hr) phenotype suggests that SPI-1 may interact with cellular and/or other viral proteins. We devised a genetic screen for suppressors of SPI-1 hr mutations by first introducing a mutation into SPI-1 (T309R) at residue P14 of the serpin reactive center loop. The SPI-1 T309R serpin is inactive as a protease inhibitor in vitro. Introduction of the mutation into RPV leads to the same restricted hr phenotype as deletion of the SPI-1 gene. Second-site suppressors were selected by restoration of growth of the RPV SPI-1 T309R hr mutant on A549 cells. Both intragenic and extragenic suppressors of the T309R mutation were identified. One novel intragenic suppressor mutation, T309C, restored protease inhibition by SPI-1 in vitro. Extragenic suppressor mutations were mapped by a new procedure utilizing overlapping PCR products encompassing the entire genome in conjunction with marker rescue. One suppressor mutation, which also rendered the virus temperature sensitive for growth, mapped to the DNA polymerase gene (E9L). Several other suppressors mapped to gene D5R, an NTPase required for DNA replication. These results unexpectedly suggest that the host range function of SPI-1 may be associated with viral DNA replication by an as yet unknown mechanism.

Advances in Pyrazole Based Scaffold as Cyclin-Dependent Kinase 2 Inhibitors for the Treatment of Cancer

2021 ◽  
Vol 21 ◽  
Author(s):  
Jahara Shaikh ◽  
Kavitkumar Patel ◽  
Tabassum Khan
Keyword(s):  
Cell Cycle ◽  
Proliferative Activity ◽  
Docking Studies ◽  
Cyclin Dependent Kinase ◽  
Cyclin Dependent Kinases ◽  
Ic50 Value ◽  
Anti Cancer ◽  
Specific Cancer ◽  
Cell Cycle Deregulation

: The transformation of a normal cell into a tumor cell is one of the initial steps in cell cycle deregulation. The cell cycle is regulated by cyclin-dependent kinases (CDKs) that belong to the protein kinase family. CDK2 is an enchanting target for specific genotypes tumors since cyclin E is selective for CDK2 and the deregulation of specific cancer forms. Thus, CDKs inhibitor specifically CDK2/cyclin A-E has the potential to be a valid cancer target as per the currently undergoing clinical trials. Mostly pyrazole scaffolds have shown selectivity and potency for CDK2 inhibitors. This review demonstrates pyrazole and pyrazole fused with other heterocyclic rings for anti-proliferative activity. Based on the in vitro and molecular docking studies, the IC50 value of various hybrids is revealed to display the most potent analogs for CDK2 inhibition. Thus, the review emphasizes various lead analogs of pyrazole hybrids which can be found to be very potent and selective for anti-cancer drugs.

scholarly journals Structure-Based Virtual Screening: Identification of a Novel NS2B-NS3 Protease Inhibitor with Potent Antiviral Activity against Zika and Dengue Viruses

2021 ◽  
Vol 9 (3) ◽  
pp. 545
Author(s):  
Hye Jin Shin ◽  
Mi-Hwa Kim ◽  
Joo-Youn Lee ◽  
Insu Hwang ◽  
Gun Young Yoon ◽  
...  
Keyword(s):  
Virtual Screening ◽  
Protease Activity ◽  
Therapeutic Option ◽  
Denv Infection ◽  
Antiviral Drugs ◽  
Protease Inhibition ◽  
Docking Analysis ◽  
Ns3 Protease ◽  
Protease Inhibition Assay

Zika virus (ZIKV), which is associated with severe diseases in humans, has spread rapidly and globally since its emergence. ZIKV and dengue virus (DENV) are closely related, and antibody-dependent enhancement (ADE) of infection between cocirculating ZIKV and DENV may exacerbate disease. Despite these serious threats, there are currently no approved antiviral drugs against ZIKV and DENV. The NS2B-NS3 viral protease is an attractive antiviral target because it plays a pivotal role in polyprotein cleavage, which is required for viral replication. Thus, we sought to identify novel inhibitors of the NS2B-NS3 protease. To that aim, we performed structure-based virtual screening using 467,000 structurally diverse chemical compounds. Then, a fluorescence-based protease inhibition assay was used to test whether the selected candidates inhibited ZIKV protease activity. Among the 123 candidate inhibitors selected from virtual screening, compound 1 significantly inhibited ZIKV NS2B-NS3 protease activity in vitro. In addition, compound 1 effectively inhibited ZIKV and DENV infection of human cells. Molecular docking analysis suggested that compound 1 binds to the NS2B-NS3 protease of ZIKV and DENV. Thus, compound 1 could be used as a new therapeutic option for the development of more potent antiviral drugs against both ZIKV and DENV, reducing the risks of ADE.

scholarly journals Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp5 main protease

2021 ◽  
Vol 478 (13) ◽  
pp. 2499-2515 ◽  
Author(s):  
Jennifer C. Milligan ◽  
Theresa U. Zeisner ◽  
George Papageorgiou ◽  
Dhira Joshi ◽  
Christelle Soudy ◽  
...  
Keyword(s):  
Economic Effects ◽  
Calpain Inhibitor ◽  
Chemical Library ◽  
Antiviral Compounds ◽  
Main Protease ◽  
The World ◽  
Ic50 Values ◽  
Global Population ◽  
Micromolar Range

The coronavirus 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread around the world with unprecedented health and socio-economic effects for the global population. While different vaccines are now being made available, very few antiviral drugs have been approved. The main viral protease (nsp5) of SARS-CoV-2 provides an excellent target for antivirals, due to its essential and conserved function in the viral replication cycle. We have expressed, purified and developed assays for nsp5 protease activity. We screened the nsp5 protease against a custom chemical library of over 5000 characterised pharmaceuticals. We identified calpain inhibitor I and three different peptidyl fluoromethylketones (FMK) as inhibitors of nsp5 activity in vitro, with IC50 values in the low micromolar range. By altering the sequence of our peptidomimetic FMK inhibitors to better mimic the substrate sequence of nsp5, we generated an inhibitor with a subnanomolar IC50. Calpain inhibitor I inhibited viral infection in monkey-derived Vero E6 cells, with an EC50 in the low micromolar range. The most potent and commercially available peptidyl-FMK compound inhibited viral growth in Vero E6 cells to some extent, while our custom peptidyl FMK inhibitor offered a marked antiviral improvement.

scholarly journals Anti-cancer Activity of Garcinia mangostana L. and Its Derivatives in Cervical Cancer

2020 ◽  
pp. 21-30
Author(s):  
S. V. Kirthanashri ◽  
N. Ramesh Kumar ◽  
S. Chitra
Keyword(s):  
Cervical Cancer ◽  
Clonogenic Assay ◽  
Cancer Cell Line ◽  
In Vitro Study ◽  
Cervical Cancer Cell Line ◽  
Garcinia Mangostana ◽  
Ic50 Value ◽  
Anti Cancer ◽  
Cancer Activity

The Garcinia mangostana Linn, a medicinal plant commonly used in Southeast Asia. The crude extract and isolated metabolites were used to evaluate its potential anti-cancer activity which was compared to methotrexate (MTX) in cervical cancer cell line. The present study involved in using crude mangosteen (CM), γ-mangostin (γ-M), isopropyl mangostin (IPM) and Di-O-methyl mangostin (DMM) against the standard anti-neoplastic drug MTX. Cell viability and cytotoxicity by lactate dehydrogenase (LDH) were assessed. Analysis of DNA fragmentation and clonogenic assay further supports the anti-cancer activity of the drugs in HeLa cells. The IC50 value for CM, γ-M, DMM, IPM and MTX were 13.4µg, 34.84µM, 15.57µM 5.3µM and 16.05µM respectively observed in the present study. This study suggests that the G. mangostana and its derivatives have potential anti-cancer activity an in vitro study in cervical cancer.

scholarly journals Identification, Characterization and Synthesis of Natural Parasitic Cysteine Protease Inhibitors — More Potent Falcitidin Analogs

2021 ◽  
Author(s):  
Stephan Brinkmann ◽  
Sandra Semmler ◽  
Christian Kersten ◽  
Maria A. Patras ◽  
Micheal Kurz ◽  
...  
Keyword(s):  
Protease Inhibitors ◽  
Cysteine Protease ◽  
Therapeutic Option ◽  
In Silico Analysis ◽  
Biosynthetic Gene Cluster ◽  
Acyl Residue ◽  
Cysteine Protease Inhibitors ◽  
Protease Inhibition ◽  
Parasite Plasmodium Falciparum

Protease inhibitors represent a promising therapeutic option for the treatment of parasitic diseases such as malaria and human African trypanosomiasis. Falcitidin was the first member of a new class of inhibitors of falcipain 2, a cysteine protease of the malaria parasite Plasmodium falciparum. Using a metabolomics dataset of 25 Chitinophaga strains for molecular networking enabled identification of over 30 natural analogs of falcitidin. Based on MS/MS spectra, they vary in their amino acid chain length, sequence, acyl residue, and C terminal functionalization; therefore, they were grouped into the four falcitidin peptide families A-D. The isolation, characterization and absolute structure elucidation of two falcitidin-related pentapeptide aldehyde analogs by extensive MS/MS spectrometry and NMR spectroscopy in combination with advanced Marfey's analysis was in agreement with the in silico analysis of the corresponding biosynthetic gene cluster. Total synthesis of chosen pentapeptide analogs followed by in vitro testing against a panel of proteases revealed selective parasitic cysteine protease inhibition and additionally low-micromolar inhibition of α-chymotrypsin. The pentapeptides investigated here showed superior inhibitory activity compared to falcitidin.

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