| 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 |
Polθ ( IC50 = 7.6 nM )
ART812 targets DNA polymerase theta (Polθ), a key enzyme in the microhomology-mediated end joining (MMEJ) DNA repair pathway. Polθ is a multifunctional enzyme with both polymerase and helicase domains, and it is responsible for annealing microhomologous sequences and extending DNA ends during MMEJ. The compound inhibits Polθ polymerase activity with an IC50 of 7.6 nM in biochemical assays. In cell-based MMEJ assays, ART812 has an IC50 of 240 nM【25L9-L10】. By inhibiting Polθ, ART812 blocks the MMEJ repair pathway, leading to the accumulation of DNA damage and cell death in HR-deficient cancer cells. |
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| ln Vitro |
In MDA-MB-436 SHLD2 mutant cells, ART812 (0–40 μM) promotes Polθ inhibitor sensitivity [1]
ART812 demonstrates potent in vitro activity against Polθ in biochemical and cell-based assays. The compound inhibits Polθ polymerase activity with an IC50 of 7.6 nM. In cell-based MMEJ assays, ART812 has an IC50 of 240 nM【25L9-L10】. The compound has shown efficacy in inhibiting the growth of PARP-resistant BRCA1 mutant tumor cells. ART812’s activity is attributed to its selective inhibition of Polθ, which is synthetically lethal with HR deficiency. |
| ln Vivo |
In rats exhibiting MDA-MB-436 BRCA1/SHLD2-deficient tumors (volume 250-350 mm3), ART812 (100 mg/kg; administered orally daily for 76 days) showed notable tumor reduction [1]
ART812 has demonstrated in vivo efficacy in inhibiting PARP-resistant BRCA1 mutant tumor growth in rats. The compound is orally active, making it suitable for oral administration in preclinical models. Further studies would be needed to evaluate the efficacy of ART812 in other HR-deficient cancer models and to characterize its pharmacokinetic and safety profile. |
| Enzyme Assay |
The polymerase inhibitory activity of ART812 can be assessed using in vitro polymerase assays. In a typical assay, purified recombinant Polθ polymerase domain is incubated with a primer-template DNA substrate, nucleotide substrates (including radiolabeled or fluorescently labeled nucleotides), and varying concentrations of ART812. The incorporation of nucleotides into the growing DNA strand is measured, and the half-maximal inhibitory concentration (IC50) is calculated from dose-response curves. The IC50 for Polθ inhibition is 7.6 nM. The selectivity of ART812 against other DNA polymerases can be assessed using similar assays with other polymerases.
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| Cell Assay |
The cellular activity of ART812 is assessed using cell-based MMEJ reporter assays and HR-deficient cancer cell lines. In MMEJ reporter assays, cells are transfected with a reporter construct that can only be repaired by MMEJ, and the frequency of repair is measured by flow cytometry or by measuring reporter gene expression. Cells are treated with ART812 at varying concentrations, and the IC50 for inhibition of MMEJ is calculated from dose-response curves. In HR-deficient cancer cell lines, such as BRCA1/2-mutant cells, the effects of ART812 on cell viability, DNA damage, and apoptosis are assessed using MTT assays, γH2AX staining (a marker of DNA damage), and flow cytometry.
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| Animal Protocol |
ART812 has been evaluated in a rat xenograft model of BRCA1 mutant tumor growth. Rats bearing BRCA1 mutant tumors were treated with ART812 by oral administration. The compound demonstrated efficacy in inhibiting tumor growth. The dosing regimen and administration route have not been detailed in the available literature, but the compound is described as orally active.
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| ADME/Pharmacokinetics |
ART812 is orally active, suggesting that it has favorable oral bioavailability. The compound is a small molecule with a molecular weight of 461.80 g/mol. Further studies would be needed to characterize the absorption, distribution, metabolism, and excretion (ADME) properties of ART812, including plasma protein binding, clearance, half-life, and tissue distribution. The compound’s pharmacokinetic profile would be essential for determining appropriate dosing regimens for in vivo efficacy studies.
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| Toxicity/Toxicokinetics |
No detailed toxicity data for ART812 has been published in the available literature. As a research compound, ART812 has not been subjected to comprehensive toxicology studies. The mechanism of action, which involves the inhibition of Polθ and the induction of synthetic lethality in HR-deficient cells, suggests that the compound could have a manageable safety profile, with toxicity potentially limited to HR-deficient cancer cells. However, off-target effects and the potential for toxicity in normal tissues would need to be carefully assessed in future studies.
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| References | |
| Additional Infomation |
ART812 is an orally active and selective inhibitor of DNA polymerase theta (Polθ), with an IC50 of 7.6 nM in biochemical assays and 240 nM in cell-based MMEJ assays. The compound inhibits the MMEJ DNA repair pathway and is synthetically lethal with HR deficiency. ART812 has demonstrated efficacy in inhibiting PARP-resistant BRCA1 mutant tumor growth in rats. The compound is a promising therapeutic approach for HR-deficient cancers, including breast, ovarian, prostate, and pancreatic cancers, and for overcoming resistance to PARP inhibitors.
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| Molecular Formula |
C19H16CLF4N3O4
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|---|---|
| Molecular Weight |
461.79
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| Exact Mass |
461.08
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| Elemental Analysis |
C, 49.42; H, 3.49; Cl, 7.68; F, 16.46; N, 9.10; O, 13.86
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| CAS # |
2607138-82-7
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| PubChem CID |
156077039
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| Appearance |
White to yellow solid powder
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| LogP |
2.8
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
31
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| Complexity |
700
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| Defined Atom Stereocenter Count |
3
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| SMILES |
N1(C2=NC(C)=CC(C(F)(F)F)=C2)C(=O)[C@@H](O)[C@@H](O)[C@H]1C(N(C1=CC=C(F)C(Cl)=C1)C)=O
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| InChi Key |
YJJOABOMFPGRED-JYJNAYRXSA-N
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| InChi Code |
InChI=1S/C19H16ClF4N3O4/c1-8-5-9(19(22,23)24)6-13(25-8)27-14(15(28)16(29)18(27)31)17(30)26(2)10-3-4-12(21)11(20)7-10/h3-7,14-16,28-29H,1-2H3/t14-,15-,16-/m0/s1
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| Chemical Name |
(2S,3S,4S)-N-(3-chloro-4-fluorophenyl)-3,4-dihydroxy-N-methyl-1-[6-methyl-4-(trifluoromethyl)pyridin-2-yl]-5-oxopyrrolidine-2-carboxamide
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| Synonyms |
ART812
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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 : ~100 mg/mL (~216.6 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.41 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 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: ≥ 2.5 mg/mL (5.41 mM) (saturation unknown) 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. 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 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: ≥ 2.5 mg/mL (5.41 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 | 2.1655 mL | 10.8274 mL | 21.6549 mL | |
| 5 mM | 0.4331 mL | 2.1655 mL | 4.3310 mL | |
| 10 mM | 0.2165 mL | 1.0827 mL | 2.1655 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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