Foscarnet

Manufactured by Merck Group

Sourced in Italy, Switzerland, United States

Foscarnet is a laboratory product manufactured by Merck Group. It functions as an antiviral agent.

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Lab products found in correlation

Cycloheximide (chx), by Merck Group (3 mentions) Cidofovir, by Gilead Sciences (2 mentions) Ganciclovir, by Merck Group (2 mentions) Fomivirsen isis 2922, by Metabion (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Recombinant human pad2, by Cayman Chemical (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Donkey anti rabbit hrp, by GE Healthcare (1 mentions) Ecl detection kit, by Cytiva (1 mentions) Anti ha, by Merck Group (1 mentions) Actinomycin d, by Merck Group (1 mentions) Ganciclovir, by Roche (1 mentions) Acyclovir, by GlaxoSmithKline (1 mentions) Afm30a, by Cayman Chemical (1 mentions) Cay10723, by Cayman Chemical (1 mentions) Gsk199, by Cayman Chemical (1 mentions) Risedronate, by Merck Group (1 mentions) Lamivudine, by Merck Group (1 mentions) Adefovir, by Merck Group (1 mentions)

11 protocols using foscarnet

Recombinant Enzyme and Plasmid Protocols

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Recombinant human PAD2, PAD4, and Cl-A were from Cayman Chemical (Ann Arbor, USA). Cycloheximide and Foscarnet were from Sigma-Aldrich (Milan, Italy). BB-Cl-A was kindly provided by P. R. Thompson (University of Massachusetts, Medical School).
The plasmid pET30A hIFIT1-GST-His was kindly provided by A. Pichlmair (Technische Universitaet Munich, Germany). pSGIE72 IE1-encoding plasmid was used in the quantitative nucleic acid analysis. The pSicoR-CRISPR-Cas9 vector (RP-557) was kindly provided by R. J. Lebbink (UMC, Utrecht).

Griffante G., Gugliesi F., Pasquero S., Dell’Oste V., Biolatti M., Salinger A.J., Mondal S., Thompson P.R., Weerapana E., Lebbink R.J., Soppe J.A., Stamminger T., Girault V., Pichlmair A., Oroszlán G., Coen D.M., De Andrea M, & Landolfo S. (2021). Human cytomegalovirus-induced host protein citrullination is crucial for viral replication. Nature Communications, 12, 3910.

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Tetracycline-Inducible HBV Replication Assay

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The HepAD38 cell line (obtained from Dr. Christoph Seeger at Fox Chase Cancer Center, Philadelphia) supporting HBV pgRNA transcription and subsequent viral DNA replication in a tetracycline (tet)-inducible manner was maintained as previously described [63 (link)]. C3A cell line [a derivative of HepG2 (ATCC HB-8065)] (ATCC CRL-10741) was maintained in DMEM/F-12 (1:1) medium supplemented with 10% FBS (GEMINI Bio-Products), 100 U/ml penicillin, 100 μg/ml streptomycin. Entecavir (ETV) is a gift from Dr. William S. Mason at Fox Chase Cancer Center, Philadelphia [64 (link)]. Foscarnet was purchased from Sigma. Capsid assembly modulators ENAN-34017 and Bay41-4109, GLS4 and AT-61 were described previously [35 (link),65 (link)]. Myrcludex B is a gift of Dr. Stephan Urban at Heidelberg University, Germany [26 (link)]. Alpha-interferon (IFN-α) was purchased from PBL Assay Science. Rabbit anti-HBc antibody was obtained from Dako (B0586).

Guo F., Zhao Q., Sheraz M., Cheng J., Qi Y., Su Q., Cuconati A., Wei L., Du Y., Li W., Chang J, & Guo J.T. (2017). HBV core protein allosteric modulators differentially alter cccDNA biosynthesis from de novo infection and intracellular amplification pathways. PLoS Pathogens, 13(9), e1006658.

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MCMV Immediate-Early Protein Analysis

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3T3 cells were infected with MCMV-m166HA. To selectively analyze immediate-early protein expression, cycloheximide (100 µg/ml; Sigma) was added to 3T3 cells 30 min prior to infection and was let sit on cells for up to 3 h post infection. At this time, cycloheximide media was washed away and actinomycin D (2.5 µg/ml; Sigma) was added to cells for another 3 h. To block late protein expression, Foscarnet (250 µg/ml; Sigma) was added just prior to infection and was present throughout. At 48 h after infection, cell pellets were scarped and lysed in protein sample buffer (3% SDS, 2% β-mercaptoethanol, 200 mM Tris [pH 8.8], 0.5M sucrose, 5 mM EDTA), boiled for 5 min, separated by SDS-PAGE (10% gel) and transferred to nitrocellulose filters. Filters were probed with anti-HA (H6908; Sigma) at 2 µg/ml followed by detection with donkey anti-rabbit HRP (GE healthcare). Signals were detected with the ECL detection kit (Amersham).

Verma S., Loewendorf A., Wang Q., McDonald B., Redwood A, & Benedict C.A. (2014). Inhibition of the TRAIL Death Receptor by CMV Reveals Its Importance in NK Cell-Mediated Antiviral Defense. PLoS Pathogens, 10(8), e1004268.

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Evaluation of Antiviral Compounds in Cell Culture

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The following antivirals were used: acyclovir (ACV, [9-(2-hydroxyethoxymethyl)guanine]), GlaxoSmithKline, Stevenage, UK; ganciclovir (GCV, [9-(1,3-dihydroxy-2-propoxymethyl)guanine), Roche, Basel, Switzerland; foscarnet (PFA, [phosphonoformate sodium salt]), Sigma Chemicals, St. Louis, MO, USA; cidofovir (CDV, [(S)-1-(3-hydroxy-2-phosphonomethoxypropyl)cytosine]), Gilead Sciences, Foster City, CA, USA; brivudin (BVDU, [(E)-5-(2-bromovinyl)-2′-deoxyuridine]), Searle, UK; HDVD, 1-[(2S,4S-2-(hydroxymethyl)-1,3-dioxolan-4-yl]5-vinylpyrimidine-2,4(1H,3H)-dione, Dr. D. Chu (University of Georgia, Athens, GA, USA); KAY-2-41 [1-(2-deoxy-1-methyl-4-thio-β-Dribofuranosyl)thymine and KAH-39-139 [1-(2-deoxy-4-amino-4-thio-β-Dribofuranosyl)thymine], Showa University, Tokyo, Japan; HPMP-5azaC [(1-(S)-(3-hydroxy-2-(phosphonomethoxy)propyl)-5-azacytosine] and HPMPO-DAPy [(R)-2,4-diamino-3-hydroxy-6-[2-(phosphonomethoxy)propoxy])pyrimidine], Dr M. Krecmerova, Institute of Organic Chemistry and Biochemistry, Prague, Czech Republic.

Trompet E., Temblador A., Gillemot S., Topalis D., Snoeck R, & Andrei G. (2021). An MHV-68 Mutator Phenotype Mutant Virus, Confirmed by CRISPR/Cas9-Mediated Gene Editing of the Viral DNA Polymerase Gene, Shows Reduced Viral Fitness. Viruses, 13(6), 985.

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Inhibition of PAD Enzymes Impacts Viral Infection

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The PAD inhibitors Cl-amidine (Cl-A), BB-Cl-amidine (BB-Cl), GSK199, CAY10727, and AFM30a—also known as CAY10723—were obtained from Cayman Chemical (Ann Arbor). CHX and foscarnet (phosphonoformic acid, PFA) were from Sigma-Aldrich. All these compounds were reconstituted in dimethyl sulfoxide (DMSO) or ethanol accordingly to their solubility. HF4 and Thapsigargin were kindly provided by Dr. P. Thompson and Dr. M. Corazzari, respectively. Immediately before use, the inhibitors were diluted in the cell medium to their final concentrations. For the PADs inhibitors experiments, cells were pre-treated with the inhibitors for 1 h and then infected (MOI 1) by adding the virus without changing the medium. Following virus adsorption (2 h), the viral inoculum was removed, and a new medium with fresh inhibitor was added. After that, no more medium or inhibitor was added until the samples were collected.

Pasquero S., Gugliesi F., Biolatti M., Dell’Oste V., Albano C., Bajetto G., Griffante G., Trifirò L., Brugo B., Raviola S., Lacarbonara D., Yang Q., Sudeshna S., Barasa L., Haniff H., Thompson P.R., Landolfo S, & De Andrea M. (2023). Citrullination profile analysis reveals peptidylarginine deaminase 3 as an HSV-1 target to dampen the activity of candidate antiviral restriction factors. PLOS Pathogens, 19(12), e1011849.

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NMPylation Assay for Viral Enzymes

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0.5 μM nsp12 and 5 μM nsp9 were incubated with different concentrations of Risedronate (Sigma-Aldrich, Cat#PHR1888) or Foscarnet (Sigma-Aldrich, Cat#PHR1436) in the NMPylation buffer for 5 min, then 25 μM GTP and 10 μCi [α³²P]-GTP were added to start the reaction. Reactions were performed for 10 min.

Wang B., Svetlov D, & Artsimovitch I. (2021). NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic Acids Research, 49(15), 8822-8835.

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Tetracycline-regulated hepatoma cell lines for virus studies

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Chicken hepatoma cells (LMH) were cultured in DMEM/F12 medium (Mediatech) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin. dstet5 is a LMH-derived stable cell line supporting the replication of an envelope protein-deficient DHBV genome in a tetracycline dependent manner (Guo et al., 2003 (link)). HepAD38 is a human hepatoma cell (HepG2)-derived stable cell line supporting HBV genome replication in a tetracycline dependent manner (Guo et al., 2007 (link)). Both cell lines were maintained in DMEM/F12 medium (Mediatech) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 1 μg/ml tetracycline and 200 μg/ml G-418. Chicken alpha-interferon (IFN-α) was produced and titrated as described previously (Guo et al., 2003 (link)). Foscarnet (PFA), Lamivudine (LAM), and Adefovir (ADV) were purchased from Sigma. Entecavir (ETV) was provided by Dr. William S. Mason at Fox Chase Cancer Center, Philadelphia. Bay 41-4109 was provided by Dr. Lai Wei at Hepatology Institute of Peking University, China. AT-61 and DVR-23 are synthesized in house as described previously (Campagna et al., 2013 (link); Xu et al., 2010 (link)).

Zhang P., Liu F., Guo F., Zhao Q., Chang J, & Guo J.T. (2016). Characterization of novel hepadnaviral RNA species accumulated in hepatoma cells treated with viral DNA polymerase inhibitors. Antiviral research, 131, 40-48.

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Anti-HCMV Compounds Protocol

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Berberine chloride (BBR), nitazoxanide (NTZ), foscarnet (FOS), and ganciclovir (GCV) were purchased from Sigma-Aldrich. Cidofovir (CDV) was from Gilead Sciences. Fomivirsen (ISIS 2922) was synthesized by Metabion International AG. The anti-HCMV 6-aminoquinolone compound WC5 was previously described (Mercorelli et al., 2009) .

Luganini A., Mercorelli B., Messa L., Palù G., Gribaudo G, & Loregian A. (2019). The isoquinoline alkaloid berberine inhibits human cytomegalovirus replication by interfering with the viral Immediate Early-2 (IE2) protein transactivating activity. Antiviral research, 164.

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Antiviral Drugs Sourcing Protocol

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Manidipine (MND) was obtained from Selleck Chemicals. Nitazoxanide (NTZ), foscarnet (FOS), and ganciclovir (GCV) were from Sigma-Aldrich. Ribavirin (RBV; 1-D-ribofuranosyl-1,2,4-triazole-3-car-boxamide) was purchased from Roche and cidofovir (CDV, Vistide) was from Gilead Sciences. Fomivirsen (ISIS 2922) was synthesized by Metabion International AG.

Mercorelli B., Luganini A., Celegato M., Palù G., Gribaudo G, & Loregian A. (2018). Repurposing the clinically approved calcium antagonist manidipine dihydrochloride as a new early inhibitor of human cytomegalovirus targeting the Immediate-Early 2 (IE2) protein. Antiviral research, 150.

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Isolation and Activation of Mouse T Cells

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Primary T cells were isolated from BM (femur, tibia, ileac crest) of 10-week-old female C57BL/6J mice (The Jackson Laboratory) using EasySep Mouse Pan-Naive T cell Isolation kit (Stemcell Technologies). Isolated T cells were cultured in RPMI (Thermo Fisher Scientific), with 10% FBS (Atlanta Biologic) and supplemented with 50 U/mL penicillin, 50 mg/mL streptomycin (Thermo Fisher Scientific). Cells were cultured overnight with or without 4 mM Pi (NaPO₄) (MilliporeSigma; pH 7.4) as described previously (103 (link)) and then activated with a T cell stimulation cocktail (phorbol 12-myristate 13-acetate [PMA] and ionomycin, eBioscience). Jurkat cells (clone E6-1, TIB-152) were purchased from ATCC and were cultured as described for primary T cells. Cells were pretreated with inhibitors (30 minutes) and were then treated with or without 4 mM Pi followed by RNA isolation after 48 hours. Foscarnet (phosphonoformic acid) (1 mM) was purchased from MilliporeSigma and PD173074 (300 nM) and TAS-120 (50 nM) were purchased from Selleck Chemicals.

Roberts J.L., Yu M., Viggeswarapu M., Arnst J.L., Pacifici R, & Beck GR J.r. (2023). Dietary phosphorus consumption alters T cell populations, cytokine production, and bone volume in mice. JCI Insight, 8(10), e154729.

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