| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
DHODH-IN-2 specifically targets dihydroorotate dehydrogenase (DHODH), the rate-limiting enzyme in the de novo pyrimidine biosynthesis pathway. It binds to the ubiquinone-binding site of DHODH, inhibiting the conversion of dihydroorotate to orotate. This leads to depletion of the pyrimidine nucleotide pool (UMP, UDP, UTP), inhibiting DNA and RNA synthesis, and resulting in cell cycle arrest and apoptosis in rapidly dividing cells.
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| ln Vitro |
In vitro, DHODH-IN-2 inhibits human DHODH activity with an IC50 of 15 nM. It is selective for DHODH over other enzymes in the pyrimidine biosynthesis pathway. The compound inhibits the proliferation of cancer cell lines (including HCT-116, A549, and Jurkat) with IC50 values in the range of 0.5-2 µM after 72 hours of treatment. It also inhibits the replication of certain viruses, including influenza and HSV, with EC50 values around 1-5 µM.
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| ln Vivo |
In vivo, DHODH-IN-2 has demonstrated efficacy in multiple mouse models. In the HCT-116 colon cancer xenograft model, oral administration of 50 mg/kg twice daily for 21 days reduced tumor growth by 60%. In a mouse model of rheumatoid arthritis (collagen-induced arthritis), treatment with DHODH-IN-2 (30 mg/kg/day, p.o.) significantly reduced joint swelling and paw inflammation. In a mouse model of influenza, the compound reduced viral titers and improved survival.
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| Enzyme Assay |
The in vitro DHODH inhibition assay uses a colorimetric method based on the reduction of 2,6-dichlorophenolindophenol (DCPIP). Recombinant human DHODH (1 nM) is incubated with DHODH-IN-2 (0.1-1000 nM) in assay buffer (50 mM HEPES, pH 7.5, 100 mM NaCl, 0.1% Triton X-100, 10% glycerol) for 10 minutes at 25°C. The substrate dihydroorotate (500 µM) and decylubiquinone (100 µM) are added, and the reaction is initiated. DCPIP reduction is measured at 600 nm for 10 minutes. IC50 is calculated from the inhibition curve.
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| Cell Assay |
For in vitro cell-based assays, HCT-116 or A549 cancer cells are seeded in 96-well plates at 5 × 10³ cells/well. DHODH-IN-2 is dissolved in DMSO and diluted to final concentrations of 0.1-50 µM. Cells are treated for 48-72 hours. Cell viability is measured using the MTT assay. Pyrimidine nucleotide levels (UTP, CTP) are quantified by LC-MS/MS. Cell cycle distribution and apoptosis (annexin V/PI) are analyzed by flow cytometry. Uridine rescue experiments (50 µM) are performed to confirm on-target activity.
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| Animal Protocol |
For in vivo xenograft studies, female BALB/c nude mice (6-8 weeks, 18-22 g) are inoculated subcutaneously with HCT-116 cells (5 × 10⁶). When tumors reach ~100 mm³, mice are randomized into groups (n=8). DHODH-IN-2 is formulated in 0.5% CMC-Na and administered orally at 25, 50, or 100 mg/kg twice daily for 21 days. Tumor volumes and body weights are measured every three days. At sacrifice, tumors are collected for pharmacodynamic analysis (pyrimidine levels, proliferation markers).
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| ADME/Pharmacokinetics |
Pharmacokinetic studies in mice after oral administration (50 mg/kg) show a Cmax of 12 µM at 1.5 hours, with an oral bioavailability of 65%. The half-life is 4.2 hours. The compound is moderately protein-bound (85%). It is metabolized by CYP3A4 and CYP2C9. Excretion is primarily via the biliary route. The compound has a moderate volume of distribution (1.5 L/kg) and good tissue penetration.
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| Toxicity/Toxicokinetics |
In preclinical toxicity studies, DHODH-IN-2 is well-tolerated in mice. The maximum tolerated dose is >200 mg/kg (p.o.). At doses up to 100 mg/kg/day for 14 days, no significant body weight loss, hepatotoxicity, or nephrotoxicity is observed. At higher doses (150 mg/kg), mild gastrointestinal disturbances (diarrhea) are noted. The compound is not mutagenic in the Ames test. No hematological abnormalities are observed.
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| References | |
| Additional Infomation |
DHODH-IN-2 is a white to off-white powder. It is soluble in DMSO and 0.5% CMC-Na. DHODH is a validated therapeutic target for rheumatoid arthritis (e.g., leflunomide) and is being explored for cancer and viral infections. The compound is a valuable research tool for studying the role of pyrimidine metabolism in disease. It has not entered clinical trials.
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| Molecular Formula |
C18H19BRN4O2
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|---|---|
| Molecular Weight |
403.2731
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| Exact Mass |
402.069
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| CAS # |
1644156-80-8
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| PubChem CID |
118430627
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| Appearance |
White to off-white solid powder
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| LogP |
4.6
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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 |
25
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| Complexity |
409
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| Defined Atom Stereocenter Count |
0
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| SMILES |
BrC1=C([H])C([H])=C([H])C([H])=C1OC1C(=NN(C2=NC([H])=C(C([H])=N2)C([H])([H])C([H])([H])[H])C=1C([H])([H])[H])OC([H])([H])C([H])([H])[H]
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| InChi Key |
SNDPFEDGFPXNIX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H19BrN4O2/c1-4-13-10-20-18(21-11-13)23-12(3)16(17(22-23)24-5-2)25-15-9-7-6-8-14(15)19/h6-11H,4-5H2,1-3H3
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| Chemical Name |
2-[4-(2-bromophenoxy)-3-ethoxy-5-methylpyrazol-1-yl]-5-ethylpyrimidine
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| Synonyms |
DHODHIN2; DHODH IN 2
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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 : ~33.33 mg/mL (~82.65 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.20 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4797 mL | 12.3986 mL | 24.7973 mL | |
| 5 mM | 0.4959 mL | 2.4797 mL | 4.9595 mL | |
| 10 mM | 0.2480 mL | 1.2399 mL | 2.4797 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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