| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Targets |
UBE2T (E2 ubiquitin-conjugating enzyme) and FANCL (E3 ligase). UBE2T/FANCL-IN-1 is an inhibitor of the UBE2T/FANCL complex, which catalyzes the monoubiquitination of FANCD2, a critical step in the Fanconi anemia DNA repair pathway.
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| ln Vitro |
At 100 μM, UBE2T/FANCL-IN-1 (CU2) totally prevents GST-FANCLRING from being ubiquitylated. In response to HU treatment, the level of mUb-FANCD2 was lowered by UBE2T/FANCL-IN-1 (500 μM). The level of mUb-FANCD2 produced after treatment with cisplatin (10 μM) is also decreased by UBE2T/FANCL-IN-1 (500 μM) for 6, 12, and 24 hours. After 8 days, the cells were less than 50% confluent due to an even more marked reduction in cell proliferation/growth when 250 μM UBE2T/FANCL-IN-1 and 15 μM carboplatin were combined[1].
UBE2T/FANCL-IN-1 (CU2) is a potent inhibitor of UBE2T/FANCL-mediated FANCD2 monoubiquitination. It completely inhibits the ubiquitylation of GST-FANCLRING at 100 uM. At 500 uM, it reduces the level of monoubiquitinated FANCD2 (mUb-FANCD2) in response to hydroxyurea (HU) and cisplatin treatment. The combination of 250 uM UBE2T/FANCL-IN-1 with 15 uM carboplatin leads to a pronounced decrease in cell proliferation, with cells being <50% confluent after 8 days. |
| ln Vivo |
In vivo efficacy data are not detailed in the supplier literature. By sensitizing cells to DNA cross-linking agents like carboplatin, UBE2T/FANCL-IN-1 has potential applications in combination chemotherapy to overcome drug resistance. It can be used in xenograft models of cancers with an intact FA pathway to study the effect of FA pathway inhibition on tumor response to platinum-based chemotherapy.
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| Enzyme Assay |
Non-cellular assays use purified components to reconstitute the FA pathway. Recombinant UBE2T, FANCL, FANCD2, and ubiquitin-charged E1 enzyme are incubated in a reaction buffer. UBE2T/FANCL-IN-1 is added at varying concentrations (0-500 uM). The reaction mixture is incubated at 37degC for 1 hour. The ubiquitination of FANCD2 is detected by Western blotting with an anti-FANCD2 antibody. The appearance of a slower-migrating band indicates monoubiquitination, and the inhibitor concentration required to abolish this band is determined.
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| Cell Assay |
Cellular assays involve treating cancer cells (e.g., HeLa or FA pathway-proficient cells) with UBE2T/FANCL-IN-1 (0-500 uM) followed by exposure to a DNA cross-linking agent such as cisplatin or carboplatin (5-50 uM). FANCD2 monoubiquitination is assessed by Western blot. Cell proliferation is measured by MTT or colony formation assays. Synergy is assessed by calculating the combination index. A decrease in FANCD2 monoubiquitination and enhanced cell death indicates successful target inhibition.
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| Animal Protocol |
For animal studies, UBE2T/FANCL-IN-1 would likely be formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. It could be administered intraperitoneally in combination with carboplatin to mice bearing subcutaneous tumor xenografts. Tumor volume would be measured, and at study termination, tumors would be harvested for analysis of FANCD2 monoubiquitination by Western blot. The combination treatment would be expected to show greater tumor growth inhibition than carboplatin alone.
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| ADME/Pharmacokinetics |
UBE2T/FANCL-IN-1 (CU2) has a molecular weight of 357.5 g/mol and a molecular formula of C19H31N7. It is soluble in DMSO. The compound is stored at -20degC. Detailed pharmacokinetic parameters (half-life, oral bioavailability) are not reported in the supplier literature. For in vivo formulation, a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline can be used.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for UBE2T/FANCL-IN-1 are not available. It is intended for research use only and is not for human consumption. Standard safety precautions for handling research chemicals should be followed, including the use of gloves and eye protection. As an inhibitor of the Fanconi anemia pathway, there is a potential risk of bone marrow suppression if used systemically, reflecting the pathophysiology of Fanconi anemia.
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| References | |
| Additional Infomation |
This compound is also known as CU2. It is a research-grade chemical and is not an FDA-approved drug. It is the first reported small-molecule inhibitor targeting the UBE2T-FANCL interaction in the Fanconi Anemia pathway. The Fanconi Anemia pathway is critical for repairing DNA interstrand crosslinks, and its inhibition is a promising strategy to sensitize cancer cells to DNA-damaging chemotherapies like cisplatin and carboplatin. It was first described in ACS Chemical Biology in 2019.
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| Molecular Formula |
C19H31N7
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| Molecular Weight |
357.496342897415
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| Exact Mass |
357.264
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| CAS # |
1359415-02-3
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| PubChem CID |
53202415
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| Appearance |
Off-white to yellow solid powder
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| LogP |
2.3
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
26
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| Complexity |
453
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C12=NC(CN3CCCCC3C)=NN1C(N1CCN(CC)CC1)=CC(C)=N2
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| InChi Key |
FJHJBABGFHVIDS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H31N7/c1-4-23-9-11-24(12-10-23)18-13-15(2)20-19-21-17(22-26(18)19)14-25-8-6-5-7-16(25)3/h13,16H,4-12,14H2,1-3H3
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| Chemical Name |
7-(4-ethylpiperazin-1-yl)-5-methyl-2-[(2-methylpiperidin-1-yl)methyl]-[1,2,4]triazolo[1,5-a]pyrimidine
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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 | 2.7972 mL | 13.9860 mL | 27.9720 mL | |
| 5 mM | 0.5594 mL | 2.7972 mL | 5.5944 mL | |
| 10 mM | 0.2797 mL | 1.3986 mL | 2.7972 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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