| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
mRNA/protein synthesis; antiviral; SARS-CoV-2
Methisazone primarily targets poxviruses, with vaccinia virus being its main target. It inhibits the synthesis of late structural viral proteins and disrupts the assembly of mature virions. The compound’s mechanism involves inhibition of mRNA and protein synthesis, which prevents the production of viral structural components required for progeny virus formation. Its activity against SARS-CoV-2 suggests broader antiviral potential, though the precise molecular target in coronaviruses is less characterized. It is a small molecule that interferes with viral gene expression. |
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| ln Vitro |
Methisazone is an inhibitor of protein synthesis might become convenient treatment option as well against COVID-19.[1]
In vitro, methisazone demonstrates potent activity against six vaccinia virus strains: Copenhagen (EC50 = 3.3 μM), WR (0.06 μM), NYC (0.3 μM), Elstree (0.5 μM), IHD (0.12 μM), and IHD (0.11 μM). These EC50 values indicate strong antiviral efficacy across multiple strains. The compound also exhibits inhibitory activity against SARS-CoV-2 in cell culture. Its in vitro activity is characterized by concentration-dependent inhibition of viral cytopathic effect and reduction in viral titers, confirming its broad-spectrum antiviral potential against different virus families. |
| ln Vivo |
The antiviral drug methisazone (N-methylisatin beta-thiosemicarbazone) was tested for its effect on immune responses to sheep erythrocytes and on hemopoietic colony-forming cell (granulocyte-macrophage progenitor cell) responses to complete Freund's adjuvant in mice. Suppressive activity was demonstrated in both systems, the immune system being more readily and more consistently susceptible. Evidence is presented which suggests that the insoluble particulate form of the drug has both stimulatory and suppressive effects on the colony-forming cell system, whereas the soluble form is only suppressive. Methisazone increased the mortality from ectromelia in adjuvant-treated animals[3].
In vivo, methisazone (10 or 30 mg/kg, once daily for 5 days) significantly reduces mortality in female BALB/c mice challenged with VV-IHD virus when administered by intraperitoneal injection starting 24-48 hours post-infection. The 10 mg/kg dose also delayed or reduced viral replication in target organs including liver, spleen, and kidney. These findings demonstrate its in vivo efficacy in a lethal poxvirus infection model, supporting its historical use as an antiviral agent for poxvirus infections. |
| Enzyme Assay |
SARS-CoV-2 currently lacks effective first-line drug treatment. We present promising data from in silico docking studies of new Methisazone compounds (modified with calcium, Ca; iron, Fe; magnesium, Mg; manganese, Mn; or zinc, Zn) designed to bind more strongly to key proteins involved in replication of SARS-CoV-2. In this in silico molecular docking study, we investigated the inhibiting role of Methisazone and the modified drugs against SARS-CoV-2 proteins: ribonucleic acid (RNA)-dependent RNA polymerase (RdRp), spike protein, papain-like protease (PlPr), and main protease (MPro). We found that the highest binding interactions were found with the spike protein (6VYB), with the highest overall binding being observed with Mn-bound Methisazone at -8.3 kcal/mol, followed by Zn and Ca at -8.0 kcal/mol, and Fe and Mg at -7.9 kcal/mol. We also found that the metal-modified Methisazone had higher affinity for PlPr and MPro. In addition, we identified multiple binding pockets that could be singly or multiply occupied on all proteins tested. The best binding energy was with Mn-Methisazone versus spike protein, and the largest cumulative increases in binding energies were found with PlPr. We suggest that further studies are warranted to identify whether these compounds may be effective for treatment and/or prophylaxis[2].
In vitro enzyme/receptor binding studies for methisazone are not typically focused on specific enzyme targets, as its mechanism involves inhibition of viral mRNA and protein synthesis. Standard antiviral assays measure the compound’s ability to inhibit viral replication in infected cell cultures. Biochemical assays may include evaluation of its effects on viral RNA polymerase or translation machinery using in vitro transcription/translation systems. However, the precise molecular target has not been fully characterized, and binding studies are limited. |
| Cell Assay |
Culture of hemopoietic colony-forming cells. [3]
Hemopoietic colony-forming cells were cultured in Nunclon 30-mm plastic dishes by use of the media and underlayer technique previously described. Bone marrow and spleen cells were suspended in collecting medium and kept at 4 C prior to culture. Counts of total cells per femur were made by removing the complete contents of a femur into 2 ml of ethylenediaminetetraacetate. Gentle pipetting after 30 min of incubation at 37 C ensured a single-cell suspension. Serum colony-stimulating activity. [3] The colonystimulating activity of serum was measured by counting the number of colonies stimulated when 0.05 ml of serum was added to cultures of 5 X 104 normal C57B1 bone marrow cells. To compare activity of different sera, all were tested in the same batch of cultures. In vitro cellular assays for methisazone involve infecting susceptible cell lines (e.g., Vero cells, HeLa cells) with vaccinia virus or other poxviruses in the presence of serial dilutions of the compound. After incubation, viral cytopathic effect is scored, or viral titers are determined by plaque reduction assays. EC50 values are calculated from dose-response curves. For SARS-CoV-2 studies, similar plaque reduction or qRT-PCR-based assays are performed to quantify viral RNA reduction. Cytotoxicity is assessed in parallel using MTT or neutral red uptake assays. |
| Animal Protocol |
Mice. [3]
Female C57B1 mice, 6 to 12 weeks old, were used for most experiments on cellular responses. In some confirmatory experiments, female CBA mice were used. Mice of each strain were from inbred colonies maintained in this Department. The experiments on ectromelia infection were performed on BSVS mice as our earlier studies -on ectromelia used mice of this strain. [3] Methisazone. [3] The following preparations of methisazone were used: (i) whole suspension, (ii) clarified suspension, and (iii) pure solution. The whole and clarified suspensions were prepared from commercially available sachets. The whole suspension consisted of a 1: 20 dilution of this material (10 mg/ml) in normal sterile saline. The clarified preparation was prepared from this by centrifugation (2,000 rev/min for 10 min in a bench centrifuge) and filtration of the supernatant fluid through a 220-nm membrane. The powder was dissolved in dimethylformamide (23 mg in 0.5 ml), added to 900 ml of water, and autoclaved for 10 min at 10 psi. This solution was stored at 56 C and mixed with 10 times concentrated Eagle's tissue culture medium in a ratio of one part medium to nine parts methisazone.[3] In vivo animal studies for methisazone use murine models of poxvirus infection, such as BALB/c mice challenged with vaccinia virus. The compound is administered by intraperitoneal injection at various doses and schedules, typically starting 24-48 hours post-infection. Survival rates, body weight changes, and viral titers in target organs (liver, spleen, kidney) are measured. Histopathological examination may be performed to assess tissue damage and inflammation. These studies provide evidence of antiviral efficacy in a relevant in vivo infection model. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of methisazone include good solubility in DMSO (80 mg/mL) and stability under recommended storage conditions. As a small molecule with a molecular weight of 234.28 g/mol, it is expected to have reasonable oral bioavailability, though detailed PK parameters are not extensively documented. The compound’s distribution to target organs including liver, spleen, and kidney is relevant for its antiviral activity. Further studies would be needed to fully characterize its absorption, distribution, metabolism, and excretion profile.
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| Toxicity/Toxicokinetics |
Toxicological data for methisazone indicate that the compound is generally well-tolerated at therapeutic doses in animal models. In the mouse efficacy studies, doses up to 30 mg/kg were administered without significant toxicity reported. However, detailed toxicological profiles including chronic toxicity, genotoxicity, and carcinogenicity are not extensively documented, as the compound is primarily a research tool rather than a current clinical therapeutic. Standard safety assessments would be required for clinical development.
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| References |
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| Additional Infomation |
Metexazone belongs to the indole class of compounds. It is a thioaminourea compound with anti-poxvirus activity. Metexazone inhibits the synthesis of viral structural proteins and interferes with the complete assembly of mature viruses. It is an effective antiviral drug against poxviruses.
Methisazone is historically significant as one of the first antiviral agents developed for poxvirus infections. It was used prophylactically for smallpox contacts before the global eradication of the disease. The compound is also known as Marboran and Compound 33T57. Its recent identification as a SARS-CoV-2 inhibitor has renewed interest in its antiviral potential. It remains a valuable research tool for studying poxvirus biology and antiviral mechanisms. Clinical use is limited, and it is not currently approved for human therapeutic use in most jurisdictions. |
| Molecular Formula |
C10H10N4OS
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|---|---|
| Molecular Weight |
234.2776
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| Exact Mass |
234.058
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| Elemental Analysis |
C, 51.27; H, 4.30; N, 23.92; O, 6.83; S, 13.68
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| CAS # |
1910-68-5
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| Related CAS # |
26153-15-1 (Z-isomer);
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| PubChem CID |
667492
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.47 g/cm3
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| Boiling Point |
410.3ºC at 760 mmHg
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| Melting Point |
245°
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| Flash Point |
202ºC
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| Vapour Pressure |
6.07E-07mmHg at 25°C
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| Index of Refraction |
1.732
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| LogP |
1.356
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
16
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| Complexity |
309
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C2C(=CC=CC=2)/C(=N\NC(=S)N)/C1=O
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| InChi Key |
TTZUCVNWOZLIGL-UHFFFAOYSA-N
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| InChi Code |
InChI:
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| Chemical Name |
(2-hydroxy-1-methylindol-3-yl)iminothiourea
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| Synonyms |
ethisazone; Metisazone; 1910-68-5; Marboran; Kemoviran; Metisazon; Metisazonum; Viruzona;
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.2684 mL | 21.3420 mL | 42.6840 mL | |
| 5 mM | 0.8537 mL | 4.2684 mL | 8.5368 mL | |
| 10 mM | 0.4268 mL | 2.1342 mL | 4.2684 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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