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Chemical Structure| 36791-04-5 Chemical Structure| 36791-04-5

Structure of Ribavirin
CAS No.: 36791-04-5

Chemical Structure| 36791-04-5

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Ribavirin blocks nucleic acid synthesis and inhibits both RNA and DNA viruses. Ribavirin is a nucleoside antimetabolite antiviral agent.

Synonyms: Ribasphere; NSC 163039; Ribavirin, Copegus, Rebetol, Virazole, ICN-1229

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Mohini, Amrita Talukder ;

Abstract: Eukaryotic Translation Initiation Factor (eIF4E) is a 5’-mRNA cap-binding protein. Upon the binding of the 5’-mRNA cap to eIF4E, translation initiation occurs as the 40S ribosomal complex begins translating the mRNA codon sequence. The eIF4E protein is elevated in 30% of cancers and has oncogenic potential through multiple mechanisms. Several epigenetic changes and overexpression of oncogenic proteins lead to the overexpression of eIF4E. Effects of overexpression of eIF4E at the protein level can be labeled as a hallmark of cancer and tumorigenesis. For example, when eIF4E is overexpressed, it facilitates the nuclear export of many oncogenic mRNAs coding for proteins such as , Mcl-1, VEGF, and cyclin D1. The 5’-mRNA cap structure consists of a 7-methyl guanosine ring which interacts with the tryptophan residues in the binding pocket of the eIF4E protein. , a analog, is an drug that is in ongoing clinical trials for anti-cancer effects. The triphosphate metabolite of is thought to have anti-proliferative and anti-migratory effects in cancer cells in patients suffering from breast, head, and neck cancers, carcinomas, and hepatic cancer. It is proposed in various studies that one possible mechanism of action of is through direct binding and inhibition of the eIF4E protein. Thus, can block the binding of the 5’-cap of mRNA, inhibiting translation initiation. We observe that or 5'-triphosphate (RTP) does not show significant inhibition in our in vitro studies. We use fluorescence polarization (FP), tryptophan fluorescence quenching, and cell lysate pulldown assays to examine the extent of binding of eIF4E with and RTP. In the fluorescence polarization assay, we utilize the ability of an inhibitor to bind to eIF4E, competing with a fluorescently labeled cap mimic. The fluorophore (tracer) is the EDA-m7GDP-ATTO-488 molecule, designed to have binding affinity for the cap binding site on eIF4E, where the 7-methyl guanosine cap of an endogenous mRNA would bind. Upon competitive inhibition, an inhibitor is likely to displace this tracer fluorophore molecule and bind at the said site, increasing the tumbling effect and reducing fluorescence polarization. On the other hand, the tryptophan quenching assay works on the principle that any inhibitor binding at the cap site would reduce the fluorescence signal emitted by the tryptophan 56 and 102 present in the binding pocket of eIF4E. Unlike the FP assay, the quenching assay does not require a fluorophore. Moreover, in our pull-down assay, we assess whether resin beads immobilized to m7GTP and bound to eIF4E could be used to test interaction of an inhibitor with the m7GTP:eIF4E interface. Western blot is used to visualize the resultant eluates after an inhibitory test molecule such as RTP is incubated with the m7GTP: eIF4E system for sufficient time. For comparison, we use a mRNA cap mimic, 7-methyl guanosine triphosphate (m7GTP), which shows complete inhibition through binding to the cap site on eIF4E. Assay data will be presented for m7GTP, , RTP, and analogs, where we conclude that neither nor RTP showed significant inhibition in the three assays. While there have been some controversies regarding the binding of and RTP with eIF4E at the cap site, our study aims to clarify and contribute to the ongoing debate surrounding ribavirin’s mechanism of action.

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Mohini, Amrita Talukder ;

Abstract: Eukaryotic Translation Initiation Factor (eIF4E) is a 5’-mRNA cap-binding protein. Upon the binding of the 5’-mRNA cap to eIF4E, translation initiation occurs as the 40S ribosomal complex begins translating the mRNA codon sequence. The eIF4E protein is elevated in 30% of cancers and has oncogenic potential through multiple mechanisms. Several epigenetic changes and overexpression of oncogenic proteins lead to the overexpression of eIF4E. Effects of overexpression of eIF4E at the protein level can be labeled as a hallmark of cancer and tumorigenesis. For example, when eIF4E is overexpressed, it facilitates the nuclear export of many oncogenic mRNAs coding for proteins such as c-myc, Mcl-1, VEGF, and cyclin D1. The 5’-mRNA cap structure consists of a 7-methyl guanosine ring which interacts with the tryptophan residues in the binding pocket of the eIF4E protein. Ribavirin, a guanosine analog, is an antiviral drug that is in ongoing clinical trials for anti-cancer effects. The triphosphate metabolite of ribavirin is thought to have anti-proliferative and anti-migratory effects in cancer cells in patients suffering from breast, head, and neck cancers, carcinomas, and hepatic cancer. It is proposed in various studies that one possible mechanism of action of ribavirin is through direct binding and inhibition of the eIF4E protein. Thus, ribavirin can block the binding of the 5’-cap of mRNA, inhibiting translation initiation. We observe that ribavirin or ribavirin 5'-triphosphate (RTP) does not show significant inhibition in our in vitro studies. We use fluorescence polarization (FP), tryptophan fluorescence quenching, and cell lysate pulldown assays to examine the extent of binding of eIF4E with ribavirin and RTP. In the fluorescence polarization assay, we utilize the ability of an inhibitor to bind to eIF4E, competing with a fluorescently labeled cap mimic. The fluorophore (tracer) is the EDA-m7GDP-ATTO-488 molecule, designed to have binding affinity for the cap binding site on eIF4E, where the 7-methyl guanosine cap of an endogenous mRNA would bind. Upon competitive inhibition, an inhibitor is likely to displace this tracer fluorophore molecule and bind at the said site, increasing the tumbling effect and reducing fluorescence polarization. On the other hand, the tryptophan quenching assay works on the principle that any inhibitor binding at the cap site would reduce the fluorescence signal emitted by the tryptophan 56 and 102 present in the binding pocket of eIF4E. Unlike the FP assay, the quenching assay does not require a fluorophore. Moreover, in our pull-down assay, we assess whether resin beads immobilized to m7GTP and bound to eIF4E could be used to test interaction of an inhibitor with the m7GTP:eIF4E interface. Western blot is used to visualize the resultant eluates after an inhibitory test molecule such as RTP is incubated with the m7GTP: eIF4E system for sufficient time. For comparison, we use a mRNA cap mimic, 7-methyl guanosine triphosphate (m7GTP), which shows complete inhibition through binding to the cap site on eIF4E. Assay data will be presented for m7GTP, ribavirin, RTP, and analogs, where we conclude that neither ribavirin nor RTP showed significant inhibition in the three assays. While there have been some controversies regarding the binding of ribavirin and RTP with eIF4E at the cap site, our study aims to clarify and contribute to the ongoing debate surrounding ribavirin’s mechanism of action.

Purchased from AmBeed:

Kupke, Paul ; Adenugba, Akinbami ; Schemmerer, Mathias ; Bitterer, Florian ; Schlitt, Hans J. ; Geissler, Edward K. , et al.

Abstract: Hepatitis E virus (HEV) is a major cause of acute hepatitis globally. Chronic and fulminant courses are observed especially in immunocompromised transplant recipients since administration of ribavirin (RBV) does not always lead to a sustained virol. response. By in vitro stimulation of NK cells through hepatoma cell lines inoculated with a full-length HEV and treatment with RBV, we analyzed the viral replication and cell response to further elucidate the mechanism of action of RBV on immune cells, especially NK cells, in the context of HEV infection. Co-culture of HEV-infected hepatoma cells with PBMCs and treatment with RBV both resulted in a decrease in viral replication, which in combination showed an additive effect. An anal. of NK cell functions after stimulation revealed evidence of reduced cytotoxicity by decreased TRAIL and CD107a degranulation. Simultaneously, IFN-γ production was significantly increased through the IL-12R pathway. Although there was no direct effect on the IL-12R subunits, downstream events starting with TYK-2 and subsequently pSTAT4 were upregulated. In conclusion, we showed that RBV has an immunomodulatory effect on the IL-12R pathway of NK cells via TYK-2. This subsequently leads to an enhanced IFN-γ response and thus, to an additive antiviral effect in the context of an in vitro HEV infection.

Keywords: ribavirin ; hepatitis E virus ; transplantation ; immunosuppression ; NK cells

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Alternative Products

Product Details of Ribavirin

CAS No. :36791-04-5
Formula : C8H12N4O5
M.W : 244.20
SMILES Code : [C@@H]2([N]1N=C(C(N)=O)N=C1)O[C@H](CO)[C@H]([C@H]2O)O
Synonyms :
Ribasphere; NSC 163039; Ribavirin, Copegus, Rebetol, Virazole, ICN-1229
MDL No. :MFCD00058564
InChI Key :IWUCXVSUMQZMFG-AFCXAGJDSA-N
Pubchem ID :37542

Safety of Ribavirin

GHS Pictogram:
Signal Word:Danger
Hazard Statements:H302-H341-H360
Precautionary Statements:P201-P202-P280-P312-P405-P501

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
Vero E6 cells 50.30 ± 24.01 µM 48 hours To measure the inhibitory effect of Ribavirin on SARS-CoV-2 replication, showing IC50 of 50.30 ± 24.01 µM Antimicrob Agents Chemother. 2023 Jan 24;67(1):e0131522.
Vero E6 cells >500 µM 72 hours To measure the cytotoxicity of Ribavirin on Vero E6 cells, showing CC50 >500 µM Antimicrob Agents Chemother. 2023 Jan 24;67(1):e0131522.
PLC/PRF/5 cells 40 µM 96 hours To evaluate the inhibitory effect of ribavirin monotherapy on HEV growth, results showed that ribavirin monotherapy initially inhibited HEV growth, but the HEV RNA levels gradually increased over time. Viruses. 2022 Nov 3;14(11):2440.

Clinical Trial:

NCT Number Conditions Phases Recruitment Completion Date Locations
NCT00216268 Japanese Encephalitis PHASE2 UNKNOWN 2025-05-08 King George Medical University... More >>, Lucknow, UP, 226003, India Less <<
NCT02483260 Viral Hemorrhagic Fever UNKNOWN 2025-12-22 Landstuhl Regional Medical Cen... More >>ter, Landstuhl, APO Ae, 09180, Germany Less <<
NCT00002298 HIV Infections TERMINATED - -
NCT01056757 Metastatic Breast Cancer PHASE1|PHASE2 TERMINATED 2025-11-10 Jewish General Hospital, Montr... More >>eal, Quebec, H3T 1E2, Canada Less <<
NCT00001015 HIV Infections PHASE1 COMPLETED 1990-06-01 Univ of California / San Diego... More >> Treatment Ctr, San Diego, California, 921036325, United States|San Francisco AIDS Clinic / San Francisco Gen Hosp, San Francisco, California, 941102859, United States|Harvard (Massachusetts Gen Hosp), Boston, Massachusetts, 02114, United States|Bellevue Hosp / New York Univ Med Ctr, New York, New York, 10016, United States|Univ of North Carolina, Chapel Hill, North Carolina, 275997215, United States Less <<
NCT00000733 HIV Infections PHASE1 COMPLETED 1991-10-01 Charity Hosp / Tulane Univ Med... More >> School, New Orleans, Louisiana, 70112, United States|Tulane Univ School of Medicine, New Orleans, Louisiana, 70112, United States Less <<
NCT03585725 Follicular Lymphoma|Mantle Cel... More >>l Lymphoma Less << EARLY_PHASE1 TERMINATED 2020-07-22 Weill Cornell Medicine, New Yo... More >>rk, New York, 10065, United States Less <<
NCT03889106 Lassa Fever TERMINATED 2020-10-01 Kenema Government Hospital, Ke... More >>nema, Sierra Leone Less <<
NCT01309490 Malignant Solid Tumour PHASE1|PHASE2 UNKNOWN 2025-06-14 Jewish General Hospital, Montr... More >>eal, Quebec, H3T 1E2, Canada Less <<
NCT00623168 Hemorrhagic Fever With Renal S... More >>yndrome Less << PHASE2 UNKNOWN 2025-12-21 Brian Allgood Army Community H... More >>ospital (121st Combat Support), Seoul, Korea, Republic of Less <<
NCT00001729 Chronic Hepatitis C|Fibrosis|H... More >>emolytic Anemia Less << PHASE3 COMPLETED 2025-09-02 National Institute of Diabetes... More >> and Digestive and Kidney Diseases (NIDDK), Bethesda, Maryland, 20892, United States Less <<
NCT00001854 Chronic Hepatitis C PHASE4 COMPLETED 2008-01-15 National Institutes of Health ... More >>Clinical Center, 9000 Rockville Pike, Bethesda, Maryland, 20892, United States Less <<
NCT00484328 Hepatitis C PHASE4 COMPLETED 2025-12-12 University of Maastricht, Maas... More >>tricht, 6229 ER, Netherlands Less <<
NCT02558114 Hepatitis E PHASE4 TERMINATED 2025-11-18 Hospital Universitari vall d'H... More >>ebron, Barcelona, 08035, Spain Less <<
NCT02308241 Human Papillomavirus (HPV)-Rel... More >>ated Malignancies|Recurrent/Metastatic (R/M) Human Papillomavirus (HPV)-Related Malignancies Less << COMPLETED 2022-03-25 Memorial Sloan Kettering Cance... More >>r Center at Basking Ridge, Basking Ridge, New Jersey, 07939, United States|Memorial Sloan Kettering Monmouth, Middletown, New Jersey, 07748, United States|Memorial Sloan Kettering Cancer Center @ Suffolk, Commack, New York, 11725, United States|Memorial Sloan Kettering Westchester, Harrison, New York, 10604, United States|Memorial Sloan Kettering Cancer Center, New York, New York, 10065, United States|Memorial Sloan Kettering at Mercy Medical Center, Rockville Centre, New York, United States Less <<
NCT00001123 Hantavirus Pulmonary Syndrome COMPLETED 2025-11-05 NIAID/DMID/CASG Central Unit, ... More >>Birmingham, Alabama, 35294, United States Less <<

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

4.10mL

0.82mL

0.41mL

20.48mL

4.10mL

2.05mL

40.95mL

8.19mL

4.10mL

References

 

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