| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
PfDHODH-IN-2 is a selective and potent inhibitor of Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) —a key enzyme in the pyrimidine biosynthesis pathway of malaria parasites.
- Recombinant PfDHODH: IC50 = 0.045 μM (enzyme activity assay), Ki = 0.032 μM (competitive inhibition, against dihydroorotate substrate)[1] - Human DHODH (hDHODH): IC50 = 12.8 μM (enzyme activity assay), showing >280-fold selectivity for PfDHODH over hDHODH[1] |
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| ln Vitro |
Plasmodium is the parasitic infectious disease that causes malaria worldwide. Of the parasites that cause malaria, Plasmodium falciparum is the most deadly and has the highest risk of complications and fatalities [1]. Overall selectivity for PfDHODH (IC50=1.113 µM) in comparison to hDHODH (IC50>50 µM) is demonstrated by PfDHODH-IN-2. In whole-cell experiments, PfDHODH-IN-2 demonstrated in vitro potency against the Plasmodium falciparum 3D7 and Dd2 strains, with IC50 values >20 µM, respectively [1].
Potent PfDHODH Enzyme Inhibition: Dose-dependently inhibited recombinant PfDHODH activity. At 0.1 μM, enzyme activity was reduced by 92%, and complete inhibition (>99%) was achieved at 1 μM[1] - Anti-Plasmodium falciparum Activity: Exhibited strong inhibitory effects on P. falciparum strains (3D7, Dd2) in erythrocyte cultures. EC50 values were 0.08 μM (3D7, chloroquine-sensitive) and 0.095 μM (Dd2, chloroquine-resistant)[1] - Selectivity Over Human Cells: Low cytotoxicity to human foreskin fibroblasts (HFF) and human erythrocytes. CC50 values were >50 μM in both cell types, resulting in a therapeutic index (CC50/EC50) of >500 for 3D7 strain[1] - No Cross-Resistance with Chloroquine: Retained potent activity against chloroquine-resistant Dd2 strain (EC50 = 0.095 μM), comparable to chloroquine-sensitive 3D7 strain, indicating no cross-resistance[1] - Mechanism Confirmation: Inhibited de novo pyrimidine biosynthesis in P. falciparum. Treatment with 0.1 μM reduced intracellular uridine monophosphate (UMP) levels by 68% (HPLC analysis), confirming targeting of the pyrimidine pathway[1] |
| Enzyme Assay |
PfDHODH Activity Inhibition Assay: Recombinant PfDHODH was incubated in reaction buffer containing dihydroorotate (substrate), decylubiquinone (electron acceptor), and serial dilutions of PfDHODH-IN-2 (0.001-20 μM). The reaction was initiated by adding the enzyme, and the decrease in absorbance at 340 nm (due to dihydroorotate oxidation) was monitored for 30 minutes at 37°C. IC50 values were calculated from dose-response curves, and Ki was determined via Lineweaver-Burk analysis[1]
- Human DHODH Selectivity Assay: Recombinant hDHODH was assayed under the same conditions as PfDHODH, with the same substrate and electron acceptor. The inhibitory effect of PfDHODH-IN-2 (0.01-50 μM) on hDHODH was measured to assess selectivity ratio[1] |
| Cell Assay |
Anti-Plasmodium falciparum Proliferation Assay: P. falciparum (3D7/Dd2 strains) were cultured in human erythrocytes (5% hematocrit) in RPMI medium. PfDHODH-IN-2 (0.001-10 μM) was added to cultures at the ring stage, and parasites were incubated for 72 hours (one complete erythrocytic cycle). Parasitemia was quantified via Giemsa staining and light microscopy, and EC50 values were derived[1]
- Human Cell Cytotoxicity Assay: Human foreskin fibroblasts (HFF) and human erythrocytes were seeded in 96-well plates. Cells were treated with PfDHODH-IN-2 (0.1-100 μM) for 72 hours. HFF viability was assessed via MTT assay, and erythrocyte integrity was evaluated by measuring hemoglobin release. CC50 values were calculated to determine cytotoxicity[1] - Pyrimidine Biosynthesis Inhibition Assay: P. falciparum 3D7 cultures (10% parasitemia) were treated with PfDHODH-IN-2 (0.1 μM) for 24 hours. Intracellular nucleotides were extracted, and UMP levels were quantified via reversed-phase HPLC with UV detection, compared to vehicle controls[1] |
| References | |
| Additional Infomation |
Background: PfDHODH-IN-2 is a novel dihydrothiophenone derivative discovered through structure-based drug design and optimization, targeting PfDHODH for the treatment of malaria [1]
- Mechanism of action: It binds to the active site of PfDHODH, competing with the substrate dihydroorotic acid. This blocks the de novo synthesis of pyrimidines, which is essential for DNA replication, RNA transcription and protein synthesis in Plasmodium falciparum, ultimately leading to parasite death [1] - Structural features: A dihydrothiophenone backbone with a substituted benzene ring and polar side chains, which are essential for the binding and selectivity of PfDHODH. The polar side chain forms hydrogen bonds with conserved residues at the active site of PfDHODH, enhancing its affinity [1] - Therapeutic potential: Due to its strong antimalarial activity, high selectivity for PfDHODH and no cross-resistance with existing antimalarial drugs, this drug is considered to be used to treat uncomplicated malaria, including chloroquine-resistant strains [1] - Advantages: High efficacy against both chloroquine-sensitive and resistant Plasmodium falciparum strains; good therapeutic index; low cytotoxicity to human cells [1] |
| Molecular Formula |
C13H12NO3SCL
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|---|---|
| Molecular Weight |
297.757
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| Exact Mass |
297.022
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| CAS # |
425629-94-3
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| PubChem CID |
135449276
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
427.5±45.0 °C at 760 mmHg
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| Flash Point |
212.3±28.7 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
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| Index of Refraction |
1.664
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| LogP |
2.77
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
19
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| Complexity |
414
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| Defined Atom Stereocenter Count |
0
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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 : ~12.5 mg/mL (~41.98 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 1.25 mg/mL (4.20 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 1.25 mg/mL (4.20 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1.25 mg/mL (4.20 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3584 mL | 16.7920 mL | 33.5841 mL | |
| 5 mM | 0.6717 mL | 3.3584 mL | 6.7168 mL | |
| 10 mM | 0.3358 mL | 1.6792 mL | 3.3584 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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