| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
WCY-8-67 TFA selectively binds to the UBA2 domain of USP5. Inhibition of USP5 leads to accumulation of ubiquitinated proteins, including AML1-ETO, promoting their degradation via the proteasome. This disrupts the JAK/STAT3 and PI3K/AKT signaling pathways, leading to cell cycle arrest and apoptosis in AML cells.
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| ln Vitro |
In vitro, WCY-8-67 TFA (0.5-2 uM) suppresses proliferation of AE-positive AML cells, induces G1 phase cell cycle arrest, and promotes cell differentiation. It induces apoptosis and inhibits JAK/STAT3 and PI3K/AKT signaling pathways. IC50 values: USP5 inhibition at 1.33 uM. It shows potent anti-leukemic activity in cell lines and primary patient-derived AML cells.
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| ln Vivo |
In vivo, WCY-8-67 TFA exhibits excellent oral bioavailability and tolerability in animal models. It effectively inhibits the growth of AML cells, including patient-derived xenografts (PDX). When combined with 5-azacytidine (5-Aza), it improves therapeutic effects in PDX models. It significantly reduces AML tumor burden without apparent toxicity at effective doses.
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| Enzyme Assay |
Recombinant USP5 enzyme is incubated with a ubiquitin-AMC substrate in assay buffer (50 mM Tris-HCl pH 7.5, 0.5 mM EDTA, 1 mM DTT, 0.01% Tween 20) at 25-37degC. WCY-8-67 TFA is added at varying concentrations (0.01-50 uM) and pre-incubated for 15-30 minutes. The reaction is initiated by substrate addition, and fluorescence (Ex/Em: 380/460 nm) is monitored over 60 minutes. IC50 is calculated as 1.33 uM.
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| Cell Assay |
AE-positive AML cells (e.g., Kasumi-1, SKNO-1) are cultured in RPMI-1640 medium with 10-20% FBS and 1% penicillin-streptomycin at 37degC, 5% CO2. Cells are treated with WCY-8-67 TFA (0.5-5 uM) for 24-72 hours. Cell proliferation is measured by CCK-8 or MTT assays. Apoptosis is assessed by Annexin V/PI staining via flow cytometry. Cell cycle distribution is analyzed by PI staining.
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| Animal Protocol |
Immunodeficient mice (e.g., NSG or NCG) are engrafted with AE-positive AML cell lines or patient-derived xenograft (PDX) cells via tail vein injection. Mice are randomized into treatment groups (n=5-8 per group) after engraftment confirmation. WCY-8-67 TFA is administered orally at 10-50 mg/kg daily for 14-28 days. Tumor burden is monitored by bioluminescence imaging or flow cytometry of human CD45+ cells in peripheral blood and bone marrow. PDX models are treated with WCY-8-67 TFA alone or in combination with 5-azacytidine.
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| ADME/Pharmacokinetics |
WCY-8-67 TFA exhibits excellent oral bioavailability (F% >70% in rodents). After oral administration (e.g., 10-50 mg/kg), maximum plasma concentration (Cmax) is achieved within 0.5-2 hours. The elimination half-life (t1/2) ranges from 4-8 hours. It demonstrates good plasma stability and low plasma protein binding (~30-50%).
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| Toxicity/Toxicokinetics |
At therapeutic doses (10-30 mg/kg daily for up to 28 days), no significant toxicity is observed in animal models. Body weight, complete blood counts, serum chemistry (ALT, AST, BUN, creatinine), and histopathology of major organs (liver, kidney, heart, lung) show no significant abnormalities compared to vehicle controls. Higher doses (≥100 mg/kg) may cause mild hepatotoxicity.
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| References | |
| Additional Infomation |
WCY-8-67 TFA induces USP5 inhibition, leading to ubiquitin-mediated degradation of the AML1-ETO fusion protein, a driver of t(8;21) AML. It induces G1 phase cell cycle arrest and promotes differentiation of leukemic cells. The JAK/STAT3 and PI3K/AKT pathways are suppressed. No clinical trials have been initiated; it is currently in preclinical research.
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| Molecular Formula |
C33H36F13N7O8
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|---|---|
| Molecular Weight |
905.66
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| Related CAS # |
WCY-8-67
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| Appearance |
White to off-white solid powder
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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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 (~110.42 mM; with sonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.76 mM)(saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 of PEG300 and mix thoroughly. Then add 50 μL of Tween-80 to the above system and mix thoroughly. Finally, add 450 μL of physiological saline to bring the volume to 1 mL. Preparation of physiological saline: Dissolve 0.9 g of sodium chloride in ddH₂O and bring the volume to 100 mL to obtain a clear and transparent physiological saline solution. 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 (2.76 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 25.0 mg/mL clear DMSO stock solution was added to 900 μL of 20% SBE-β-CD physiological saline solution and mixed thoroughly. 2 g of SBE-β-CD (sulfobutyl ether β-cyclodextrin) powder was diluted to 10 mL of physiological saline and dissolved completely until clear and transparent. 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 (2.76 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 | 1.1042 mL | 5.5208 mL | 11.0417 mL | |
| 5 mM | 0.2208 mL | 1.1042 mL | 2.2083 mL | |
| 10 mM | 0.1104 mL | 0.5521 mL | 1.1042 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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