Inhibition of herpes simplex virus-1 infection by MBZM-N-IBT: in silico and in vitro studies

Abstract Introduction The emergence of drug resistance and cross-resistance to existing drugs has warranted the development of new antivirals for Herpes simplex viruses (HSV). Hence, we have designed this study to evaluate the anti-viral activity of 1-[(2-methyl benzimidazole-1-yl) methyl]-2-oxo-indolin-3-ylidene] amino] thiourea (MBZM-N-IBT), against HSV-1. Method Molecular docking was performed to assess the affinity of MBZM-N-IBT for HSV-1 targets. This was validated by plaque assay, estimation of RNA and protein levels as well as time of addition experiments in vitro. Result Molecular docking analysis suggested the inhibitory capacity of MBZM-N-IBT against HSV-1. This was supported by the abrogation of the HSV-1 infectious viral particle formation with the IC50 value of 3.619 µM. Viral mRNA levels were also reduced by 72% and 84% for UL9 and gC respectively. MBZM-N-IBT also reduced the protein synthesis for gC and ICP8 significantly. While mRNA of ICP8 was not significantly affected, its protein synthesis was reduced by 47%. The time of addition experiment revealed the capacity of MBZM-N-IBT to inhibit HSV-1 at early as well as late stages of infection in the Vero cells. Similar effect of MBZM-N-IBT was also noticed in the Raw 264.7 and BHK 21 cells after HSV-1 infection. Supported by the in silico data, this can be attributed to possible interference with multiple HSV targets including the ICP8, ICP27, UL42, UL25, UL15 and gB proteins. Conclusion These results along with the lack of acute oral toxicity and significant anti-inflammatory effects suggest its suitability for further evaluation as a non-nucleoside inhibitor of HSV.

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Characterization of Herpes Simplex Viruses Selected in Culture for Resistance to Penciclovir or Acyclovir

Journal of Virology ◽

10.1128/jvi.75.4.1761-1769.2001 ◽

2001 ◽

Vol 75 (4) ◽

pp. 1761-1769 ◽

Cited By ~ 48

Author(s):

Robert T. Sarisky ◽

Matthew R. Quail ◽

Philip E. Clark ◽

Tammy T. Nguyen ◽

Wendy S. Halsey ◽

...

Keyword(s):

Herpes Simplex ◽

Sequence Similarity ◽

Cross Resistance ◽

Phenotypic Analysis ◽

Herpes Simplex Viruses ◽

Infected Cells ◽

Simplex Virus ◽

Hsv 1

ABSTRACT Penciclovir (PCV), an antiherpesvirus agent in the same class as acyclovir (ACV), is phosphorylated in herpes simplex virus (HSV)-infected cells by the viral thymidine kinase (TK). Resistance to ACV has been mapped to mutations within either the TK or the DNA polymerase gene. An identical activation pathway, the similarity in mode of action, and the invariant cross-resistance of TK-negative mutants argue that the mechanisms of resistance to PCV and ACV are likely to be analogous. A total of 48 HSV type 1 (HSV-1) and HSV-2 isolates were selected after passage in the presence of increasing concentrations of PCV or ACV in MRC-5 cells. Phenotypic analysis suggested these isolates were deficient in TK activity. Moreover, sequencing of the TK genes from ACV-selected mutants identified two homopolymeric G-C nucleotide stretches as putative hot spots, thereby confirming previous reports examining Acvr clinical isolates. Surprisingly, mutations identified in PCV-selected mutants were generally not in these regions but distributed throughout the TK gene and at similar frequencies of occurrence within A-T or G-C nucleotides, regardless of virus type. Furthermore, HSV-1 isolates selected in the presence of ACV commonly included frameshift mutations, while PCV-selected HSV-1 mutants contained mostly nonconservative amino acid changes. Data from this panel of laboratory isolates show that Pcvr mutants share cross-resistance and only limited sequence similarity with HSV mutants identified following ACV selection. Subtle differences between PCV and ACV in the interaction with viral TK or polymerase may account for the different spectra of genotypes observed for the two sets of mutants.

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Characterization of mutants of herpes simplex viruses, types 1 and 2, that produce fragments of the thymidine kinase polypeptide

Canadian Journal of Microbiology ◽

10.1139/m83-063 ◽

1983 ◽

Vol 29 (4) ◽

pp. 385-393

Author(s):

Timothy M. -P. Block ◽

Nancy J. Kuhn ◽

Karen A. Kustas ◽

William A. Held ◽

Kenneth Gross ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Thymidine Kinase ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

In Vitro Translation ◽

Herpes Simplex Viruses ◽

Simplex Virus ◽

Hsv 1

Seven tk− mutants of herpes simplex virus, type 2 (HSV-2), and three tk− mutants of herpes simplex virus, type 1 (HSV-1), were isolated which did not produce the thymidine kinase (TK) polypeptides but formed smaller polypeptides not seen in wild-type infected cells. Positive TK mRNA selection by hybridization to the cloned tk genes followed by in vitro translation identified the TK polypeptides. Comparisons of the products of partial proteolysis of the polypeptides of four HSV-2 and two HSV-1 tk− mutants to those of the parental TK polypeptides indicated that, in each case, the novel polypeptide was a fragment of the TK polypeptide, showing that these mutants have defects in the structural gene for tk. HSV-2 mutants of this sort have not been previously described. They and the HSV-1 mutants are similar to HSV-1 mutants reported previously. In addition, it was found that TK mRNA was present early in infection but was absent late in infection, suggesting that the shutoff of TK synthesis is due to message degradation. Also, HSV-2 TK mRNA did not hybridize to the cloned HSV-1 tk gene indicating that these genes have extensively diverged.

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Review of Whole Plant Extracts With Activity Against Herpes Simplex Viruses In Vitro and In Vivo

Journal of Evidence-Based Integrative Medicine ◽

10.1177/2515690x20978394 ◽

2021 ◽

Vol 26 ◽

pp. 2515690X2097839

Author(s):

Anna Garber ◽

Lianna Barnard ◽

Chris Pickrell

Keyword(s):

Herpes Simplex ◽

Plant Extracts ◽

Animal Cell ◽

Model Design ◽

Herpes Simplex Viruses ◽

Whole Plant ◽

Antiviral Medications ◽

Hsv 1

Herpes simplex viruses, HSV-1 and HSV-2, are highly contagious and cause lifelong, latent infections with recurrent outbreaks of oral and/or genital lesions. No cure exists for HSV-1 or HSV-2 infections, but antiviral medications are commonly used to prevent and treat outbreaks. Resistance to antivirals has begun to emerge, placing an importance on finding new and effective therapies for prophylaxis and treatment of HSV outbreaks. Botanicals may be effective HSV therapies as the constituents they contain act through a variety of mechanisms, potentially making the development of antiviral resistance more challenging. A wide variety of plants from different regions in the world have been studied for antiviral activity against HSV-1 and/or HSV-2 and showed efficacy of varying degrees. The purpose of this review is to summarize research conducted on whole plant extracts against HSV-1 and/or HSV-2 in vitro and in vivo. The majority of the research reviewed was conducted in vitro using animal cell lines, and some studies used an animal model design. Also summarized are a limited number of human trials conducted using botanical therapies on HSV lesions.

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Expression of Human Cytomegalovirus IE1 Leads to Accumulation of Mono-SUMOylated PML That Is Protected from Degradation by Herpes Simplex Virus 1 ICP0

Journal of Virology ◽

10.1128/jvi.01452-18 ◽

2018 ◽

Vol 92 (23) ◽

Cited By ~ 2

Author(s):

Wangheng Hou ◽

Ruth Cruz-Cosme ◽

Fayuan Wen ◽

Jin-Hyun Ahn ◽

Inez Reeves ◽

...

Keyword(s):

Herpes Simplex ◽

Herpes Simplex Virus 1 ◽

Western Blot Assay ◽

Herpes Simplex Viruses ◽

Link Type ◽

Nuclear Domain ◽

Simplex Virus ◽

Human Cmv ◽

Hsv 1

ABSTRACTTo countermeasure the host cellular intrinsic defense, cytomegalovirus (CMV) and herpes simplex viruses (HSV) have evolved the ability to disperse nuclear domain 10 (ND10, aka PML body). However, mechanisms underlying their action on ND10 differ. HSV infection produces ICP0, which degrades the ND10-forming protein PML. Human CMV (HCMV) infection expresses IE1 that deSUMOylates PML to result in dispersion of ND10. It has been demonstrated that HSV ICP0 degraded only the SUMOylated PML, so we hypothesized that HCMV IE1 can protect PML from degradation by ICP0. HCMV IE1-expressing cell lines (U-251 MG-IE1 and HELF-IE1) were used for infection of HSV-1 or transfection of ICP0-expressing plasmid. Multilabeling by immunocytochemistry assay and protein examination by Western blot assay were performed to determine the resultant fate of PML caused by ICP0 in the presence or absence of HCMV IE1. Here, we report that deSUMOylation of human PML (hPML) by HCMV IE1 was incomplete, as mono-SUMOylated PML remained in the IE1-expressing cells, which is consistent with the report by E. M. Schilling, M. Scherer, N. Reuter, J. Schweininger, et al. (J Virol 91:e02049-16, 2017,https://doi.org/10.1128/JVI.02049-16). As expected, we found that IE1 protected PML from degradation by ICP0 or HSV-1 infection. Anin vitrostudy found that IE1 with mutation of L174P failed to deSUMOylate PML and did not protect PML from degradation by ICP0; hence, we conclude that the deSUMOylation of PML is important for IE1 to protect PML from degradation by ICP0. In addition, we revealed that murine CMV failed to deSUMOylate and to protect the HSV-mediated degradation of hPML, and that HCMV failed to deSUMOylate and protect the HSV-mediated degradation of mouse PML. However, IE1-expressing cells did not enhance wild-type HSV-1 replication but significantly increased ICP0-defective HSV-1 replication at a low multiplicity of infection. Therefore, our results uncovered a host-virus functional interaction at the posttranslational level.IMPORTANCEOur finding that HCMV IE1 protected hPML from degradation by HSV ICP0 is important, because the PML body (aka ND10) is believed to be the first line of host intrinsic defense against herpesviral infection. How the infected viruses overcome the nuclear defensive structure (PML body) has not been fully understood. Herpesviral proteins, ICP0 of HSV and IE1 of CMV, have been identified to interact with PML. Here, we report that HCMV IE1 incompletely deSUMOylated PML, resulting in the mono-SUMOylated PML, which is consistent with the report of Schilling et al. (J Virol 91:e02049-16, 2017,https://doi.org/10.1128/JVI.02049-16). The mono-SUMOylated PML was subjected to degradation by HSV ICP0. However, it was protected by IE1 from degradation by ICP0 or HSV-1 infection. In contrast, IE1 with L174P mutation lost the function of deSUMOylating PML and failed to protect the degradation of the mono-SUMOylated PML. Whether the mono-SUMOylated PML has any defensive function against viral infection will be further investigated.

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In vitro antiviral activity of BanLec against herpes simplex viruses type 1 and 2

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v15i1.42320 ◽

2020 ◽

Vol 15 (1) ◽

pp. 11-18

Author(s):

Arisha Taj Mahaboob Batcha ◽

Ashish Wadhwani ◽

Gowri Subramaniam

Keyword(s):

Herpes Simplex ◽

Antiviral Activity ◽

Selectivity Index ◽

Antiviral Compound ◽

Herpes Simplex Viruses ◽

Antiviral Activities ◽

Simplex Virus ◽

Hsv 1

The present study evaluates the antiviral activity of banana lectin (BanLec) against herpes simplex virus type 1 and 2 (HSV-1 and HSV-2). Lectin was isolated from the ripen pulp of bananas (Musa paradisiaca). The study showed that lectin exhibited hemagglutination activity towards human erythrocytes A, B, AB and O group. The molecular weight of BanLec using SDS gel-electrophoresis was found to be 14,000-30,000 Da. Cytotoxicity of BanLec on the Vero cell lines showed an inhibitory concentration of 172.7 µg/mL. BanLec was virucidal and showed no cytotoxicity at the concentration tested. The lectin showed a dose-dependent antiviral activities, inhibiting HSV-1 by 16.0 µg/mL with selectivity index 10.8 and HSV-2 inhibition by 67.7 µg/mL with selectivity index 2.6. These results corroborate that BanLec could be a rich source of potential antiviral compound for HSV-1 when compared to HSV-2.

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Nectin2α (PRR2α or HveB) and Nectin2δ Are Low-Efficiency Mediators for Entry of Herpes Simplex Virus Mutants Carrying the Leu25Pro Substitution in Glycoprotein D

Journal of Virology ◽

10.1128/jvi.74.3.1267-1274.2000 ◽

2000 ◽

Vol 74 (3) ◽

pp. 1267-1274 ◽

Cited By ~ 99

Author(s):

Marc Lopez ◽

Francesca Cocchi ◽

Laura Menotti ◽

Elisa Avitabile ◽

Patrice Dubreuil ◽

...

Keyword(s):

Herpes Simplex ◽

Immunoglobulin Superfamily ◽

Soluble Form ◽

Glycoprotein D ◽

Herpes Simplex Viruses ◽

Low Efficiency ◽

Simplex Virus ◽

Herpesvirus Entry Mediator ◽

Hsv 1

ABSTRACT The receptors for entry of herpes simplex viruses 1 and 2 (HSV-1 and -2), widely expressed in human cell lines, are members of a subset of the immunoglobulin superfamily exemplified by herpesvirus entry mediator C (HveC) and the herpesvirus immunoglobulin-like receptor (HIgR). This report focuses on two members of this subset, herpesvirus entry mediator B (HveB), recently designated nectin2/PRR2α, and its splice variant isoform, nectin2/PRR2δ. Nectin2α and -δ share the ectodomain but differ in the transmembrane and cytoplasmic regions. HveB was reported to enable entry of HSV-1 carrying mutations in glycoprotein D (gD) and of HSV-2, but not of wild-type (wt) HSV-1. We report that (i) both nectin2α and -δ served as receptors for the entry of HSV-1 mutant viruses HSV-1(U10) and -(U21) and AP7r that carry the Leu25Pro substitution in gD but not for HSV-1 mutants U30 and R5000 that carry the Ser140 or Ala185 substitution in gD. All of these mutants were able to overcome the block to entry mediated by expression of wt gD. (ii) Infection of cells expressing nectin2α or -δ required exposure to multiplicities of infection about 100-fold higher than those required to infect cells expressing HveC or HIgR. (iii) gD from HSV-1(U21) bound in vitro soluble forms of nectin2. The association was weaker than that to the soluble form of HveC/HIgR. Binding of wt HSV-1 gD to soluble nectin2 was not detectable. (iv) A major region of nectin2 functional in virus entry mapped to the V domain, located at the N terminus.

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The Antiviral and Antimalarial Drug Repurposing in Quest of Chemotherapeutics to Combat COVID-19 Utilizing Structure-Based Molecular Docking

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207323999200824115536 ◽

2020 ◽

Vol 23 ◽

Author(s):

Sisir Nandi ◽

Mohit Kumar ◽

Mridula Saxena ◽

Anil Kumar Saxena

Keyword(s):

Molecular Docking ◽

In Silico ◽

Drug Repurposing ◽

Clinical Settings ◽

The Novel ◽

In Silico Docking ◽

Biochemical Mechanisms ◽

Novel Coronavirus ◽

In Silico Molecular Docking

Background: The novel coronavirus disease (COVID-19) is caused by a new strain (SARS-CoV-2) erupted in 2019. Nowadays, it is a great threat that claims uncountable lives worldwide. There is no specific chemotherapeutics developed yet to combat COVID-19. Therefore, scientists have been devoted in the quest of the medicine that can cure COVID- 19. Objective: Existing antivirals such as ASC09/ritonavir, lopinavir/ritonavir with or without umifenovir in combination with antimalarial chloroquine or hydroxychloroquine have been repurposed to fight the current coronavirus epidemic. But exact biochemical mechanisms of these drugs towards COVID-19 have not been discovered to date. Method: In-silico molecular docking can predict the mode of binding to sort out the existing chemotherapeutics having a potential affinity towards inhibition of the COVID-19 target. An attempt has been made in the present work to carry out docking analyses of 34 drugs including antivirals and antimalarials to explain explicitly the mode of interactions of these ligands towards the COVID-19protease target. Results: 13 compounds having good binding affinity have been predicted towards protease binding inhibition of COVID-19. Conclusion: Our in silico docking results have been confirmed by current reports from clinical settings through the citation of suitable experimental in vitro data available in the published literature.

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In Silico Appraisal, Synthesis, Antibacterial Screening and DNA Cleavage for 1,2,5-thiadiazole Derivative

Current Computer - Aided Drug Design ◽

10.2174/1573409915666190206142756 ◽

2019 ◽

Vol 15 (5) ◽

pp. 445-455 ◽

Cited By ~ 6

Author(s):

Suraj N. Mali ◽

Sudhir Sawant ◽

Hemchandra K. Chaudhari ◽

Mustapha C. Mandewale

Keyword(s):

Molecular Docking ◽

Dna Cleavage ◽

In Silico ◽

Oral Absorption ◽

Inhibition Mechanism ◽

Hydrogen Binding ◽

Docking Score ◽

Antibacterial Screening ◽

Mycobacteria Tuberculosis

Background: : Thiadiazole not only acts as “hydrogen binding domain” and “two-electron donor system” but also as constrained pharmacophore. Methods:: The maleate salt of 2-((2-hydroxy-3-((4-morpholino-1, 2,5-thiadiazol-3-yl) oxy) propyl) amino)- 2-methylpropan-1-ol (TML-Hydroxy)(4) has been synthesized. This methodology involves preparation of 4-morpholino-1, 2,5-thiadiazol-3-ol by hydroxylation of 4-(4-chloro-1, 2,5-thiadiazol-3-yl) morpholine followed by condensation with 2-(chloromethyl) oxirane to afford 4-(4-(oxiran-2-ylmethoxy)-1,2,5-thiadiazol- 3-yl) morpholine. Oxirane ring of this compound was opened by treating with 2-amino-2-methyl propan-1- ol to afford the target compound TML-Hydroxy. Structures of the synthesized compounds have been elucidated by NMR, MASS, FTIR spectroscopy. Results: : The DSC study clearly showed that the compound 4-maleate salt is crystalline in nature. In vitro antibacterial inhibition and little potential for DNA cleavage of the compound 4 were explored. We extended our study to explore the inhibition mechanism by conducting molecular docking, ADMET and molecular dynamics analysis by using Schrödinger. The molecular docking for compound 4 showed better interactions with target 3IVX with docking score of -8.508 kcal/mol with respect to standard ciprofloxacin (docking score= -3.879 kcal/mol). TML-Hydroxy was obtained in silico as non-carcinogenic and non-AMES toxic with good percent human oral absorption profile (69.639%). TML-Hydroxy showed the moderate inhibition against Mycobacteria tuberculosis with MIC 25.00 μg/mL as well as moderate inhibition against S. aureus, Bacillus sps, K. Pneumoniae and E. coli species. Conclusion: : In view of the importance of the 1,2,5-thiadiazole moiety involved, this study would pave the way for future development of more effective analogs for applications in medicinal field.

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Virtual Screening, Molecular docking and in-silico ADME-Tox analysis for identification of potential main protease (Mpro) enzyme inhibitors

Anti-Infective Agents ◽

10.2174/2211352518999201208201854 ◽

2020 ◽

Vol 18 ◽

Author(s):

Debadash Panigrahi ◽

Ganesh Prasad Mishra

Keyword(s):

Molecular Docking ◽

Virtual Screening ◽

In Silico ◽

Data Bank ◽

Future Research ◽

Reference Compound ◽

Unexpected Death ◽

Main Protease ◽

In Silico Admet

Objective:: Recent pandemic caused by SARS-CoV-2 described in Wuhan China in December-2019 spread widely almost all the countries of the world. Corona virus (COVID-19) is causing the unexpected death of many peoples and severe economic loss in several countries. Virtual screening based on molecular docking, drug-likeness prediction, and in silico ADMET study has become an effective tool for the identification of small molecules as novel antiviral drugs to treat diseases. Methods:: In the current study, virtual screening was performed through molecular docking for identifying potent inhibitors against Mpro enzyme from the ZINC library for the possible treatment of COVID-19 pandemic. Interestingly, some compounds are identified as possible anti-covid-19 agents for future research. 350 compounds were screened based on their similarity score with reference compound X77 from ZINC data bank and were subjected to docking with crystal structure available of Mpro enzyme. These compounds were then filtered by their in silico ADME-Tox and drug-likeness prediction values. Result:: Out of these 350 screened compounds, 10 compounds were selected based on their docking score and best docked pose in comparison to the reference compound X77. In silico ADME-Tox and drug likeliness predictions of the top compounds were performed and found to be excellent results. All the 10 screened compounds showed significant binding pose with the target enzyme main protease (Mpro) enzyme and satisfactory pharmacokinetic and toxicological properties. Conclusion:: Based on results we can suggest that the identified compounds may be considered for therapeutic development against the COVID-19 virus and can be further evaluated for in vitro activity, preclinical, clinical studies and formulated in a suitable dosage form to maximize their bioavailability.

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Cellular miR-101-1 Reduces Efficiently the Replication of HSV-1 in HeLa Cells

Intervirology ◽

10.1159/000512956 ◽

2021 ◽

Vol 64 (2) ◽

pp. 88-95

Author(s):

Bahar Sadegh Ehdaei ◽

Ahmad Pirouzmand ◽

Mehdi Shabani ◽

Arezoo Mirzaei ◽

Sharareh Moghim

Keyword(s):

Hela Cells ◽

Plaque Assay ◽

Viral Gene Expression ◽

Viral Gene ◽

Herpes Simplex Viruses ◽

Viral Progeny ◽

Immunosuppressed Patients ◽

Health Complications ◽

Hsv 1

Introduction: Herpes simplex viruses (HSVs) are widely distributed in the human population. HSV type 1 (HSV-1) is responsible for a spectrum of diseases, ranging from gingivostomatitis to keratoconjunctivitis, and encephalitis. The HSVs establish latent infections in nerve cells, and recurrences are common. Their frequent reactivation in elderly and immunosuppressed patients causes serious health complications. Objectives: Due to the growing resistance to its main drug, acyclovir, alternative treatments with different mechanisms of action are required. MicroRNAs regulate host and viral gene expression posttranscriptionally. Previous studies reported that mir-101-2 expression has widely participated in the regulation of HSV-1 replication. In this study, we investigate the effect of hsa-miR-101-1 in the replication of HSV-1. Methods: We found that transfection of miR-101-1 into HeLa cells could reduce effectively HSV-1 replication using plaque assay and real-time PCR methods. Results: We showed that overexpression of miR-10-1 produced less viral progeny and manifested a weaker cytopathic effect, without affecting cell viability. Discussion/Conclusion: This result can give us new insights into the control of HSV-1 infections.

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