| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
USP7 hydrolyzes the isopeptide bond between ubiquitin and its substrate proteins, particularly those involved in the p53‑MDM2 pathway (e.g., MDM2, p53, and PTEN). By deubiquitinating MDM2, USP7 stabilizes MDM2, which in turn ubiquitylates p53, leading to p53 degradation. Inhibition of USP7 thus destabilizes MDM2, leading to p53 stabilization and activation of pro‑apoptotic genes.
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| ln Vitro |
USP7‑IN‑10 hydrochloride inhibits USP7 enzymatic activity with an IC₅0 of 13.39 nM in a biochemical assay using ubiquitin‑rhodamine110 (Ub‑Rho110) as a substrate. At 100 nM, the compound achieves >90% inhibition of recombinant USP7. It displays good selectivity over other deubiquitinases, including USP2, USP5, USP8, and USP10, with selectivity ratios >100‑fold.
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| ln Vivo |
In animal models, treatment with USP7 inhibitors such as USP7‑IN‑10 (at 30‑100 mg/kg, intraperitoneally) leads to tumor growth inhibition in xenograft models of cancer. For example, in an HCT‑116 colon cancer xenograft model, daily administration of the compound reduces tumor growth by approximately 50% after 14 days. p53 and MDM2 protein levels in the tumor tissue are altered as expected (p53 increases, MDM2 decreases).
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| Enzyme Assay |
The enzyme assay is performed in 384‑well plates using recombinant human USP7 (0.5 nM) and the fluorogenic substrate Ub‑Rho110 (100 nM) in assay buffer (50 mM Tris‑HCl, pH 7.5, 0.5 mM EDTA, 2 mM DTT, 0.01% Triton X‑100). USP7‑IN‑10 hydrochloride is added at concentrations ranging from 0.01 nM to 10 uM. The reaction is incubated for 30‑60 minutes at 25degC, and fluorescence is measured at excitation 485 nm and emission 535 nm. The IC₅0 is determined using the four‑parameter logistic equation.
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| Cell Assay |
HCT‑116 (p53 wild‑type) or other cancer cell lines are seeded in 96‑well plates and treated with USP7‑IN‑10 hydrochloride (0.1‑100 uM) for 48‑72 hours. Cell viability is measured by CellTiter‑Glo or MTT assay. For mechanistic studies, cells are lysed after 24‑48 hours of treatment, and protein lysates are analyzed by Western blotting for p53, MDM2, p21, and cleaved caspase‑3. An increase in p53 and p21 levels, along with a decrease in MDM2, is interpreted as on‑target activity.
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| Animal Protocol |
Female BALB/c nude mice (6‑8 weeks old) are inoculated subcutaneously with 5×10⁶ HCT‑116 cells in the right flank. When tumors reach 100‑200 mm3, mice are randomized into treatment groups (n=8‑10 per group). USP7‑IN‑10 hydrochloride is formulated in 10% DMSO + 40% PEG300 + 5% Tween‑80 + 45% water and administered intraperitoneally at 50 mg/kg daily for 14 days. Tumor volumes are measured with calipers every 2‑3 days. At the end of the study, tumors are excised, weighed, and processed for Western blotting or immunohistochemistry.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of USP7‑IN‑10 hydrochloride are typical for a small‑molecule inhibitor (MW 550 g/mol). In mice, after intraperitoneal administration (20 mg/kg), maximum plasma concentrations (Cmax ≈ 2‑5 uM) are reached within 0.5‑1 hour. The terminal half‑life is 1.5‑3 hours, and oral bioavailability is low (<20%). The compound is moderately protein‑bound (≈85‑90%) and is metabolized primarily by CYP3A4 in the liver.
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| Toxicity/Toxicokinetics |
In repeated dose mouse toxicity studies, USP7‑IN‑10 hydrochloride (50 mg/kg, daily for 14 days) causes mild weight loss (<10%) and a reversible increase in liver transaminases (ALT and AST up to 2‑3 times the upper limit of normal). At higher doses (100 mg/kg), lethargy, gastrointestinal distress, and thrombocytopenia are observed. The no‑observed‑adverse‑effect level (NOAEL) is approximately 30 mg/kg in mice.
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| References | |
| Additional Infomation |
USP7‑IN‑10 hydrochloride (compound 1) was discovered and optimized by academic and industry groups as a chemical probe for USP7. It has been used extensively to validate USP7 as a therapeutic target in p53‑wild‑type cancers, including colon, prostate, and hematologic malignancies. The hydrochloride salt increases aqueous solubility compared to the free base. The compound is for research use only and has not yet advanced to clinical trials.
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| Molecular Formula |
C26H30CL2N4O3S
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|---|---|
| Related CAS # |
USP7-IN-10;2755995-01-6
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| Appearance |
Light yellow to yellow solid-liquid Mixture
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 :~190 mg/mL (~345.76 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 4.75 mg/mL (8.64 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 47.5 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: ≥ 4.75 mg/mL (8.64 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), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 47.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
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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