| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
DHODH-IN-9 targets human dihydroorotate dehydrogenase (DHODH), a flavin-dependent mitochondrial enzyme that catalyzes the fourth and rate-limiting step of de novo pyrimidine biosynthesis, converting dihydroorotate to orotate. DHODH is essential for the proliferation of rapidly dividing cells, including virus-infected cells, as they require sufficient pyrimidine nucleotides for DNA and RNA synthesis. By inhibiting DHODH, the compound depletes intracellular pyrimidine pools, thereby limiting viral replication. DHODH-IN-9 is an azine-bearing analog and demonstrates antiviral activity with a pMIC₅₀ of 7.4. The compound targets influenza virus as well as DHODH, indicating broad-spectrum antiviral potential.
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| ln Vitro |
The equivalent pyridazine homologue of 3-chloro-6-cyclopropylpyridazine is DHODH-IN-9 (compound 10k). The 5-cyclopyridinyl group analogues' antiviral method of action is essentially unaltered, and it somewhat resembles the 5-ethylpyrimidinyl group homologs' antiviral mode of action. Of these analogues, DHODH-IN-9 (compound 10k) is the superior antiviral medication [1].
In vitro, DHODH-IN-9 is a human dihydroorotate dehydrogenase inhibitor with an antiviral effect. The compound demonstrates antiviral activity with a pMIC₅₀ of 7.4. It targets influenza virus and DHODH. The antiviral activity is attributed to the inhibition of DHODH, which depletes pyrimidine nucleotide pools and limits viral replication. DHODH-IN-9 is an azine-bearing analog and serves as a valuable tool for studying DHODH biology and developing antiviral therapies, particularly against influenza virus and other DHODH-dependent pathogens. The compound's activity is concentration-dependent, with efficacy observed at nanomolar to low micromolar concentrations. |
| ln Vivo |
In vivo, DHODH-IN-9 has not been extensively reported in the available literature. As a DHODH inhibitor with antiviral activity against influenza virus, the compound would be expected to demonstrate efficacy in animal models of influenza infection. DHODH inhibitors have been studied in various preclinical models for their antiviral and immunomodulatory properties. The compound's pMIC₅₀ of 7.4 indicates potent antiviral activity in cell-based assays. However, detailed in vivo efficacy, pharmacokinetic, and toxicological data for DHODH-IN-9 require further investigation from primary research publications.
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| Enzyme Assay |
In vitro enzyme/receptor binding (non-cellular) assays for DHODH-IN-9 involve measuring inhibition of human recombinant DHODH enzymatic activity. The assay typically uses a colorimetric method detecting the conversion of dihydroorotate to orotate coupled with the reduction of the electron acceptor (e.g., 2,6-dichloroindophenol, DCIP). The enzyme is incubated with varying concentrations of the compound, substrate (dihydroorotate), and coenzyme (ubiquinone or decylubiquinone) in assay buffer (typically 50 mM Tris-HCl, pH 8.0, with 0.1% Triton X-100). The reaction is monitored spectrophotometrically by following the decrease in absorbance at 600 nm (DCIP reduction). IC₅₀ values are calculated from concentration-response curves.
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| Cell Assay |
In vitro cellular experiments with DHODH-IN-9 are performed using virus-infected cell lines to assess antiviral activity. Influenza virus replication assays are typically conducted in MDCK or A549 cells. Cells are cultured in appropriate medium (e.g., DMEM with 10% FBS) and infected with influenza virus at a suitable multiplicity of infection (MOI). Following virus adsorption, cells are treated with varying concentrations of DHODH-IN-9 for 24-72 hours. Viral replication is quantified by measuring viral RNA levels using RT-qPCR, viral protein expression by Western blot or immunofluorescence, or viral titers by plaque assay or TCID₅₀. The pMIC₅₀ of 7.4 indicates potent inhibition of viral replication. Cytotoxicity is assessed in parallel using MTT or CellTiter-Glo assays to determine the selectivity index.
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| Animal Protocol |
In vivo animal studies with DHODH-IN-9 have not been extensively reported. For antiviral studies against influenza, standard in vivo models include mouse models of influenza infection. Mice are infected with influenza virus via intranasal inoculation and treated with DHODH-IN-9 via oral, intraperitoneal, or intravenous administration at various doses. Efficacy is assessed by survival rates, weight loss, viral burden in lung tissues, and histopathological examination of lung inflammation. Pharmacodynamic markers such as pyrimidine nucleotide levels in lung tissues may be measured to confirm target engagement. The compound's antiviral activity with pMIC₅₀ of 7.4 suggests potential for in vivo efficacy.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of DHODH-IN-9 are not extensively characterized. The compound has molecular formula C₂₁H₂₃FN₄O and molecular weight 366.43. As a small molecule (molecular weight 366.43) containing a fluorine atom, it is expected to have reasonable oral bioavailability and metabolic stability. The compound's azine-bearing structure may influence its physicochemical properties. Storage: follow manufacturer's guidelines. Detailed pharmacokinetic parameters including half-life, clearance, and bioavailability require further investigation from primary research publications.
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| Toxicity/Toxicokinetics |
Toxicological information for DHODH-IN-9 is not extensively detailed in the available literature. As a research compound with enzyme inhibitory and antiviral activity, it should be handled with appropriate safety precautions. Standard safety guidelines for handling potent pharmaceutical compounds apply, including use of personal protective equipment (gloves, safety goggles, lab coat), working in a well-ventilated area, and proper chemical waste disposal. The compound is intended for research use only and is not approved for human therapeutic use. Cytotoxicity studies in cell-based assays help establish the therapeutic window and selectivity index of the compound.
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| References | |
| Additional Infomation |
DHODH-IN-9 (CAS 1644156-41-1) is an azine-bearing analog and a human dihydroorotate dehydrogenase (DHODH) inhibitor. The compound has molecular formula C₂₁H₂₃FN₄O and molecular weight 366.43. DHODH-IN-9 has an antiviral effect with a pMIC₅₀ of 7.4. It targets DHODH, an enzyme crucial in the de novo pyrimidine biosynthesis pathway that catalyzes the fourth step of pyrimidine nucleotide synthesis, converting dihydroorotate to orotate. DHODH-IN-9 is also known as Compound 10k and hDHODH-IN-7. The compound's antiviral activity is attributed to its inhibition of DHODH, which depletes pyrimidine nucleotide pools and limits viral replication.
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| Molecular Formula |
C21H23FN4O
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| Molecular Weight |
366.4319
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| Exact Mass |
366.185
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| CAS # |
1644156-41-1
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| PubChem CID |
118430504
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
4.9
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
27
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| Complexity |
481
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
HNZNVWVVPQVTKE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H23FN4O/c1-13(2)27-20-18(10-16-6-4-5-7-19(16)22)14(3)26(25-20)21-23-11-17(12-24-21)15-8-9-15/h4-7,11-13,15H,8-10H2,1-3H3
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| Chemical Name |
5-cyclopropyl-2-[4-[(2-fluorophenyl)methyl]-5-methyl-3-propan-2-yloxypyrazol-1-yl]pyrimidine
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| Synonyms |
DHODHIN9; DHODH IN 9
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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 | 2.7290 mL | 13.6452 mL | 27.2903 mL | |
| 5 mM | 0.5458 mL | 2.7290 mL | 5.4581 mL | |
| 10 mM | 0.2729 mL | 1.3645 mL | 2.7290 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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