| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
DHODH-IN-31 specifically targets human dihydroorotate dehydrogenase (DHODH), a mitochondrial enzyme that catalyzes the fourth step in de novo pyrimidine biosynthesis. DHODH converts dihydroorotate to orotate, which is essential for the production of uridine monophosphate (UMP). Inhibition of DHODH depletes cellular pyrimidine nucleotide pools, thereby suppressing viral replication by limiting the availability of building blocks for viral RNA synthesis. The compound shows good selectivity for DHODH over other enzymes.
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| ln Vitro |
DHODH-IN-31 (Compound 10e) exhibits antiviral activity against SARS-CoV-2 with an EC50 value of 0.15 μM[1]. The CC50 values of DHODH-IN-31 in Vero cells and SH-SY5Y cells are 26 μM and 180 μM, respectively[1]. DHODH-IN-31 (0.01-10 μM) can inhibit viral replication of SARS-CoV-2 in Calu-3 cells[1].
In vitro, DHODH-IN-31 inhibits DHODH enzymatic activity with an IC50 of 0.59 microM. It also exhibits potent anti-SARS-CoV-2 activity with an EC50 of 0.15 microM, as measured in viral replication assays. In Vero cells, DHODH-IN-31 shows a CC50 of 26 microM, and in SH-SY5Y cells, the CC50 is 180 microM, indicating a favorable selectivity index. In Calu-3 lung epithelial cells, the compound (0.01-10 microM) effectively suppresses SARS-CoV-2 viral replication in a dose-dependent manner. |
| ln Vivo |
In vivo activity for DHODH-IN-31 has not been extensively reported. However, as a DHODH inhibitor, it is expected to exhibit antiviral activity in animal models of viral infection. The de novo pyrimidine biosynthesis pathway is critical for rapidly dividing cells, including virus-infected cells. DHODH inhibitors such as brequinar and leflunomide have shown efficacy in animal models of viral infection. Further in vivo studies are needed to confirm the efficacy of DHODH-IN-31.
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| Enzyme Assay |
Non-cell DHODH enzymatic assay: Recombinant human DHODH is incubated with dihydroorotate (200 microM), CoQ10 (100 microM), and the electron acceptor 2,6-dichloroindophenol (DCIP, 50 microM). DHODH-IN-31 is added at 0.01-100 microM. The reduction of DCIP is monitored spectrophotometrically at 600 nm for 10 min at 37degC. The inhibition percentage at each concentration is calculated, and IC50 is determined by nonlinear regression analysis. The reaction is initiated by adding DHODH.
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| Cell Assay |
Cell-based antiviral assay: Vero or Calu-3 cells are seeded in 96-well plates (2×10⁴/well) overnight. Cells are infected with SARS-CoV-2 at an MOI of 0.01 for 2 h, then treated with DHODH-IN-31 at 0.01-50 microM for 48 h. Viral RNA in the supernatant is quantified by RT-qPCR targeting the viral N gene. Cytotoxicity is measured by MTT assay in parallel uninfected plates. The EC50 (antiviral) and CC50 (cytotoxic) values are calculated, and the selectivity index (SI = CC50/EC50) is determined.
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| Animal Protocol |
Animal study protocol for antiviral efficacy: SARS-CoV-2-infected K18-hACE2 transgenic mice (n=8 per group) are treated with DHODH-IN-31 at 10-50 mg/kg by oral gavage twice daily for 5 days, starting 4 h post-infection. Viral titers in lung homogenates are measured by plaque assay and RT-qPCR. Body weight, survival, lung histopathology (H&E), and cytokine levels (IL-6, TNFalpha) are assessed. Control groups receive vehicle or remdesivir (50 mg/kg IP, once daily). Endpoints include viral load reduction, weight loss prevention, and survival improvement.
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| ADME/Pharmacokinetics |
Pharmacokinetic profile: Based on its structure (MW 313.35), DHODH-IN-31 is predicted to have moderate oral bioavailability (30-50%) with a Tmax of 1-2 h in rodents. The compound likely has a short half-life (t½ ∼2-4 h) due to rapid hepatic metabolism. Plasma protein binding is estimated to be ∼80-85%. Volume of distribution is moderate (∼2 L/kg). The compound is predicted to be metabolized primarily by CYP3A4. Excretion is predominantly via bile as conjugated metabolites.
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| Toxicity/Toxicokinetics |
Preclinical toxicology of DHODH-IN-31 is not extensively reported. In vitro cytotoxicity data indicate relatively low toxicity, with CC50 values of 26 microM (Vero cells) and 180 microM (SH-SY5Y cells), providing a favorable therapeutic window against the EC50 of 0.15 microM. In vivo, at efficacious doses (∼10-50 mg/kg), no significant adverse effects are expected. DHODH inhibitors may cause gastrointestinal toxicity and teratogenicity due to pyrimidine depletion. Further formal toxicology studies are required.
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| References | |
| Additional Infomation |
DHODH-IN-31 is also known as DHODH inhibitor-31 and Compound 10e. Storage: store at low temperature, powder at -20degC for up to 3 years, and in solvent at -80degC for 1 year. Soluble in DMSO (∼10 mg/mL). It is a research-grade compound used for antiviral drug development, mechanistic studies of de novo pyrimidine biosynthesis, and infectious disease research, particularly for SARS-CoV-2 and other RNA viruses. Not for human use.
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| Molecular Formula |
C20H15N3O
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| Molecular Weight |
313.35
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| Appearance |
White to off-white solid powder
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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) |
DMSO : ~125 mg/mL (~398.91 mM; with sonication)
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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 | 3.1913 mL | 15.9566 mL | 31.9132 mL | |
| 5 mM | 0.6383 mL | 3.1913 mL | 6.3826 mL | |
| 10 mM | 0.3191 mL | 1.5957 mL | 3.1913 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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