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| 1mg |
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| 5mg |
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| Targets |
The compound targets the mRNAs encoding the immediate-early 2 (IE2) protein of human cytomegalovirus (HCMV). Fomivirsen is a 21-mer antisense phosphorothioate oligonucleotide that is complementary to the IE2 mRNA sequence. By binding to this mRNA, the compound triggers RNase H-mediated degradation of the mRNA, preventing the synthesis of the IE2 protein, which is essential for viral replication. This sequence-specific mechanism makes it a targeted antiviral agent with minimal off-target effects.
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| ln Vitro |
Via the antisense mechanism, antisense oligonucleotides impede viral replication by complementing the messenger RNA of the main early region protein of the human cytomegalovirus [1]. The 21-mer phosphorothioate ASO Fomiwesen sodium has a CpG motif close to its 5' terminator. Its sequence, 5'-GCG TTT GCT CTT CTT CTT GCG-3', results in mRNA regulation by an RNase H-mediated mechanism[2]. Human fibroblasts (MRC-5 cells) and pigment epithelial (RPE) cells' CMV replication is dose-regulated by fomiwesen sodium. RPE cells have an average IC50 of 0.03 μM, while MRC5 cells have an average IC50 of 0.2 μM[1].
Fomivirsen demonstrates antiviral activity against human CMV (HCMV) clinical isolates in cell-based assays. In retinal pigment epithelial (RPE) cells and MRC-5 fibroblasts, fomivirsen effectively inhibits HCMV AD169 replication, with activity comparable to or superior to the standard antiviral drug ganciclovir in certain cell types. The compound binds to and degrades the mRNAs encoding the immediate-early 2 protein, thereby inhibiting virus proliferation. |
| ln Vivo |
Specific in vivo activity data are not detailed, but fomivirsen was approved for the local treatment of CMV retinitis. Intravitreal injection of fomivirsen into the eye achieves high local concentrations of the drug, effectively suppressing CMV replication in the retina. In clinical use, fomivirsen was administered by intravitreal injection (i.e., directly into the vitreous humor of the eye) rather than systemically, as systemic administration of phosphorothioate oligonucleotides can be associated with side effects and the drug has poor oral bioavailability.
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| Enzyme Assay |
Fomivirsen is an antisense oligonucleotide, not an enzyme inhibitor, so standard enzyme assays are not applicable. Instead, the compound's activity is measured by its ability to reduce target mRNA levels. In cell-based assays (see below), CMV-infected cells are treated with fomivirsen, and the levels of IE2 mRNA and protein are measured by qRT-PCR and Western blot, respectively. Viral replication is assessed by plaque reduction assay or by measuring viral DNA by qPCR.
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| Cell Assay |
The antiviral activity of fomivirsen is assessed in cell-based assays using CMV-susceptible cells such as human foreskin fibroblasts (HFF) or retinal pigment epithelial (RPE) cells. Cells are seeded in 96-well plates and infected with HCMV (e.g., AD169 strain or clinical isolates) at a low multiplicity of infection (MOI = 0.01-0.1). After viral adsorption, varying concentrations of fomivirsen (0.1-100 microM) are added to the culture medium. Cells are incubated for 3-7 days, and viral replication is quantified by (1) plaque reduction assay, (2) qPCR quantification of viral DNA, (3) measurement of viral IE2 mRNA by qRT-PCR, or (4) detection of viral antigens (e.g., IE2 or pp65) by immunostaining. The EC50 for inhibition of viral replication is calculated.
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| Animal Protocol |
In vivo antiviral activity and toxicology studies for fomivirsen were performed in the context of its regulatory approval. A typical animal study would use the rabbit model of CMV retinitis or the murine CMV model. Female New Zealand White rabbits are injected intravitreally with fomivirsen (e.g., 50-250 microg/eye) 24 hours before or after intravitreal injection of HCMV (0.1 mL of 10⁵ PFU). At various time points, animals are euthanized, and eyes are enucleated for histopathological examination of retinal damage, measurement of viral load by qPCR, and assessment of drug distribution.
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| ADME/Pharmacokinetics |
Fomivirsen is administered by intravitreal injection, which results in high local concentrations in the vitreous and retina with minimal systemic absorption. The half-life of fomivirsen in the vitreous humor is approximately 24-36 hours, and the drug is cleared via the aqueous humor outflow and enzymatic degradation. Systemic bioavailability is very low (<5%). The compound is not orally bioavailable and does not cross the blood-brain barrier or blood-retinal barrier well.
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| Toxicity/Toxicokinetics |
Fomivirsen is toxic when administered systemically. Systemic administration of phosphorothioate oligonucleotides can cause complement activation, thrombocytopenia, elevation of liver enzymes, and pro-inflammatory effects. Therefore, the approved route of administration is intravitreal injection, which allows for local treatment with minimal systemic exposure. Local toxicity associated with intravitreal injection can include uveitis (inflammation of the uveal tract), vitritis, and increased intraocular pressure. The therapeutic index is favorable when administered locally.
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| References | |
| Additional Infomation |
Fomivirsen (brand name: Vitravene) was approved by the U.S. FDA in 1998 for the local treatment of CMV retinitis in immunocompromised patients, including those with AIDS. It was the first antisense oligonucleotide drug to receive regulatory approval. However, due to the introduction of highly active antiretroviral therapy (HAART) for HIV/AIDS and the consequent decline in incidence of CMV retinitis, as well as competition from more convenient oral antiviral agents, fomivirsen was discontinued in 2002 and is no longer commercially available. Nevertheless, it remains a landmark compound in the history of nucleic acid therapeutics and is available as a research-grade chemical for laboratory use.
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| Molecular Formula |
C204H243N63NA20O114P20S20
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|---|---|
| Molecular Weight |
7122.03656983376
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| CAS # |
160369-77-7
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| Related CAS # |
Fomivirsen;144245-52-3
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| Appearance |
White to off-white solid powder
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| SMILES |
S=P([O-])(OC[C@@H]1[C@H](C[C@H](N2C=NC3C(NC(N)=NC2=3)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(N=C(C=C2)N)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(N=C(C=C2)N)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(N=C(C=C2)N)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(N=C(C=C2)N)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(NC(C(C)=C2)=O)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C=NC3C(NC(N)=NC2=3)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C(N=C(C=C2)N)=O)O1)OP([O-])(OC[C@@H]1[C@H](C[C@H](N2C=NC3C(NC(N)=NC2=3)=O)O1)O)=S)=S)=S)=S)=S)=S)=S)=S)=S)=S)=S)=S)=S)=S)O[C@H]1C[C@H](N2C(NC(C(C)=C2)=O)=O)O[C@@H]1COP([O-])(O[C@H]1C[C@H](N2C(NC(C(C)=C2)=O)=O)O[C@@H]1COP([O-])(O[C@H]1C[C@H](N2C(NC(C(C)=C2)=O)=O)O[C@@H]1COP([O-])(O[C@H]1C[C@H](N2C=NC3C(NC(N)=NC2=3)=O)O[C@@H]1COP([O-])(O[C@H]1C[C@H](N2C(N=C(C=C2)N)=O)O[C@@H]1COP([O-])(O[C@@H]1[C@@H](CO)O[C@H](C1)N1C=NC2C(NC(N)=NC1=2)=O)=S)=S)=S)=S)=S.[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+]
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ~100 mg/mL (~14.04 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (14.04 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.1404 mL | 0.7020 mL | 1.4041 mL | |
| 5 mM | 0.0281 mL | 0.1404 mL | 0.2808 mL | |
| 10 mM | 0.0140 mL | 0.0702 mL | 0.1404 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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