| 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 |
USP1/UAF1(IC50= 544 nM)
The primary target of SJB2-043 is ubiquitin-specific protease 1 (USP1), which forms a complex with the Usp1-associated Factor 1 (UAF1). USP1 is a deubiquitinating enzyme involved in DNA damage repair, particularly in the Fanconi anemia pathway and translesion synthesis. By inhibiting USP1, SJB2-043 prevents the deubiquitination of its substrates, leading to the degradation of ID proteins (ID1, ID2, ID3), which are crucial regulators of cell differentiation and proliferation. This inhibition disrupts DNA repair mechanisms and promotes apoptosis in cancer cells. |
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| ln Vitro |
At a micromolar drug concentration, SJB2-043 causes a dose-dependent decrease in the levels of ubiquitin-specific protease 1 (USP1) and concurrent degradation of the inhibitor of DNA-binding-1 (ID1) protein in K562 cells. In K562 cells, SJB2-043 also results in a reduction in the amounts of other ID proteins, specifically ID2 and ID3. With an EC50 of roughly 1.07 μM, SJB2-043 reduces the number of viable K562 cells in a dose-dependent manner. Moreover, K562 cells undergo dose-dependent apoptosis when exposed to SJB2-043[1].
SJB2-043 demonstrates potent in vitro inhibition of the USP1 enzyme with an IC50 value of 544.0 nM. It effectively inhibits the activity of the native USP1/UAF1 complex. Additionally, it promotes the degradation of ID1, ID2, and ID3 proteins, which are key regulators of cell differentiation and proliferation. The compound also inhibits the growth of the leukemic cell line K562. Furthermore, SJB2-043 exhibits antiviral activity by reducing SARS-CoV-2 induced cytotoxicity in VERO-6 cells at 10 µM. |
| ln Vivo |
SJB2-043 demonstrates antiviral activity in vivo by reducing SARS-CoV-2 induced cytotoxicity in VERO-6 cells, with a 3.45% inhibition observed after 48 hours of exposure to 0.01 MOI of the virus. Its primary application is in cancer research, where it is used to study the role of USP1 in tumor growth and DNA repair mechanisms. The compound's ability to inhibit USP1 and promote ID protein degradation makes it a valuable tool for validating USP1 as a therapeutic target in cancer and other diseases.
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| Enzyme Assay |
Utilizing ubiquitin-AMC (Ub-7-amido-4methylcoumarin) as the substrate, the in vitro enzymatic assays are carried out in a reaction buffer that also contains 20 mM HEPES-KOH (pH 7.8), 20 mM NaCl, 0.1 mg/mL ovalbumin, 0.5 mM EDTA, and 10 mM dithiothreitol. The FluoStar Galaxy Fluorometer is used to measure the fluorescence. The proteins are incubated with Ub-vinylsulfone (VS) at a final concentration of 0.5 μM for 45 minutes at 30°C in the Ub-vinylsulfone (VS) assay. This is followed by the immunoblotting analysis[1].
The in vitro enzyme assay for SJB2-043 involves measuring its inhibition of the USP1/UAF1 deubiquitinating activity. The compound is incubated with the native USP1/UAF1 complex and a ubiquitinated substrate. The deubiquitination activity is measured, and the IC50 is determined from dose-response curves. SJB2-043 inhibits the USP1 enzyme with an IC50 value of 544.0 nM. It also inhibits the labeling of USP1 by Ub-VS (ubiquitin vinyl sulfone) in a dose-dependent manner, indicating its ability to block the active site of the enzyme. |
| Cell Assay |
Leukemic cell lines are cultured in RPMI 1640 medium with penicillin/streptomycin and 10% fetal bovine serum added. Hela and U2OS cells are cultured in DMEM that has been enhanced with penicillin/streptomycin and 10% fetal bovine serum. The USP1 inhibitor C527 and its derivatives, such as SJB2-043, are synthesized, and high-performance liquid chromatography is used to confirm the purity. Samples from primary human AML patients are obtained from the DFCI leukemia program in accordance with approved protocols. For 24-72 hours, cells are treated with DMSO or USP1 inhibitors (such as SJB2-043) in the proper medium. The MTT assay, Cell TiterGlo reagent, or Trypan blue staining are used to calculate the viable cell counts.Using flow cytometry and Annexin V and 7AAD staining, the apoptotic cells are identified. The cells are resuspended in 45 μL of PBS plus 5 μL of Benzidine stain solution (0.2% in 0.5 M glacial acetic acid, 3% H2O2) after being twice washed with PBS for the staining process. By using light microscopy, the benzidine-positive cells are found after 45 minutes of incubation at room temperature[1].
In vitro cellular assays for SJB2-043 typically use leukemic cell lines such as K562. Cells are treated with varying concentrations of the compound, and the effects on cell proliferation, ID protein levels, and apoptosis are assessed. The compound promotes ID1 degradation and inhibits cell growth. It also decreases the levels of ID2 and ID3 proteins. Additionally, the compound's antiviral activity is assessed in VERO-6 cells infected with SARS-CoV-2, where it reduces viral-induced cytotoxicity. |
| Animal Protocol |
In vivo animal studies for SJB2-043 involve mouse models of cancer, such as leukemia xenografts. Mice bearing tumors are treated with the compound via oral gavage or intraperitoneal injection at doses determined from preclinical studies. Tumor growth, ID protein levels, and survival are monitored. The compound's ability to inhibit USP1 and sensitize tumors to DNA-damaging agents is evaluated. However, detailed published in vivo efficacy data are limited, and further studies are needed to fully characterize its therapeutic potential.
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| ADME/Pharmacokinetics |
SJB2-043 is a small molecule with good cell permeability. It is soluble in DMSO and should be stored at -20°C for long-term preservation as a powder and at -80°C in solvent. Its molecular weight is approximately 275.26 g/mol. For in vivo studies, it can be formulated in suitable vehicles such as 10% DMSO + 90% corn oil or other standard formulations. Its pharmacokinetic properties, including oral bioavailability, half-life, and tissue distribution, are not extensively reported and would need to be characterized for therapeutic applications.
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| Toxicity/Toxicokinetics |
Toxicological data for SJB2-043 are limited as it is a research compound. It is intended for research use only and is not for human therapeutic applications. Standard safety precautions should be followed when handling, including the use of personal protective equipment. Its potential toxicity may be related to its mechanism of action, as USP1 inhibition could affect normal DNA repair in healthy cells. For detailed toxicity information, specialized safety data sheets should be consulted.
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| References | |
| Additional Infomation |
SJB2-043 is a potent USP1 inhibitor (IC50 = 544 nM) and a C527 analog. It promotes ID1 degradation and inhibits the growth of K562 leukemic cells. It also decreases ID2 and ID3 levels and exhibits antiviral activity against SARS-CoV-2. The compound has the CAS number 63388-44-3. It is a research tool for studying USP1 in cancer and DNA repair, and it is not FDA-approved for therapeutic use.
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| Molecular Formula |
C17H9NO3
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|---|---|
| Molecular Weight |
275.26
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| Exact Mass |
275.058
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| Elemental Analysis |
C, 74.18; H, 3.30; N, 5.09; O, 17.44
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| CAS # |
63388-44-3
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| Related CAS # |
63388-44-3;
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| PubChem CID |
509070
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| Appearance |
Green to dark green solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
495.0±48.0 °C at 760 mmHg
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| Flash Point |
253.2±29.6 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.656
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| LogP |
4.6
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
21
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| Complexity |
443
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1C(C2C([H])=C([H])C([H])=C([H])C=2[H])=NC2C(C3=C([H])C([H])=C([H])C([H])=C3C(C1=2)=O)=O
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| InChi Key |
CMYQQADDUUDCCA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H9NO3/c19-14-11-8-4-5-9-12(11)15(20)16-13(14)18-17(21-16)10-6-2-1-3-7-10/h1-9H
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| Chemical Name |
2-Phenyl-naphth[2,3-d]oxazole-4,9-dione
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| Synonyms |
SJB2-043 SJB2 043 SJB2043
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| HS Tariff Code |
2934.99.03.00
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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 : 1.5~ 3.33 mg/mL ( 5.44~12.10 mM )
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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 | 3.6329 mL | 18.1646 mL | 36.3293 mL | |
| 5 mM | 0.7266 mL | 3.6329 mL | 7.2659 mL | |
| 10 mM | 0.3633 mL | 1.8165 mL | 3.6329 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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