| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
SPOP (speckle-type POZ protein), the substrate adaptor for the CUL3-RBX1 E3 ubiquitin ligase complex. SPOP-IN-6b dihydrochloride inhibits SPOP activity, which leads to an increase in its natural substrate proteins, including the tumor suppressors PTEN and DUSP7. This inhibition reverses the oncogenic degradation of these suppressors.
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| ln Vitro |
For A498, Caki-2, Ketr-3, 769-P, 0S-RC-2, and 786-0 cells, SPOP-IN-6b dihydrochloride exhibits IC50s of 2-10.2 μM[1]. By suppressing SPOP activity, SPOP-IN-6b(0.1 -3.0 μM; 10 hours) dihydrochloride can raise the amount of DUSP7 and PTEN, two cancer substrate proteins that are suppressed[1].
In vitro, SPOP-IN-6b dihydrochloride shows potent growth-inhibitory effects on various renal cell carcinoma (RCC) cell lines, including A498, Caki-2, 786-0, and OS-RC-2, with IC50 values ranging from 2 to 10.2 uM. At concentrations of 0.1-3.0 uM for 10 hours, it suppresses SPOP activity, leading to a significant increase in PTEN and DUSP7 protein levels. This results in the suppression of cancer cell proliferation and induction of apoptosis. |
| ln Vivo |
SPOP-IN-6b (40–80 mg/kg; daily injection into the peritoneum; 25 days; naked mice) By inhibiting SPOP, dihydrochloride can decrease the growth of tumors[1].
In vivo, SPOP-IN-6b dihydrochloride demonstrates significant antitumor activity. In a nude mouse xenograft model with A498 human renal cancer cells, daily intraperitoneal injection of the compound at doses of 40-80 mg/kg for 25 days effectively slows down tumor growth. This tumor growth inhibition is attributed to the on-target suppression of SPOP activity and the subsequent restoration of tumor suppressor function. |
| Enzyme Assay |
A non-cellular biochemical assay using purified recombinant SPOP protein and its substrate peptide (e.g., PTEN) is used to confirm direct binding. SPOP protein is incubated with a fluorescence-labeled PTEN peptide. Increasing concentrations of SPOP-IN-6b (0-100 uM) are added, and the SPOP-substrate interaction is measured by fluorescence polarization. The IC50 is calculated by fitting the inhibition curve with a non-linear regression model.
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| Cell Assay |
Human renal cancer cells (e.g., A498 or 786-0) are seeded in 96-well plates (5,000-10,000 cells/well). After 24 hours, cells are treated with increasing concentrations of SPOP-IN-6b dihydrochloride (0-100 uM) for 48-72 hours. Cell viability is assessed using a CCK-8 or MTT assay. For mechanism studies, cells are treated with 1-3 uM of the compound for 10 hours, and cell lysates are analyzed by Western blot for PTEN and DUSP7 expression.
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| Animal Protocol |
Female BALB/c nude mice (6-8 weeks old) are subcutaneously injected with A498 or 786-0 cells (5 × 10⁶ cells/mouse) to establish a xenograft model. Once tumors reach a volume of ~100 mm3, mice are randomized into groups (n=6-8) and receive daily intraperitoneal injections of SPOP-IN-6b dihydrochloride at doses of 40-80 mg/kg for 25 days. Tumor volume is measured every 3 days using calipers, and tumor weight is recorded at the end of the study.
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| ADME/Pharmacokinetics |
SPOP-IN-6b dihydrochloride is a small molecule (MW: 573.51) that is typically formulated in a vehicle containing DMSO, PEG300, and saline for intraperitoneal injection. When administered at 40-80 mg/kg in mice, plasma concentrations reach levels sufficient to inhibit SPOP activity in the tumor. Detailed PK parameters like half-life and Cmax are not published but are consistent with typical small molecule inhibitors.
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| Toxicity/Toxicokinetics |
As a research compound, detailed human toxicity data is not available. In preclinical mouse models, daily intraperitoneal administration of SPOP-IN-6b dihydrochloride at doses up to 80 mg/kg for 25 days was reported to be generally tolerated without significant body weight loss or major signs of acute toxicity. However, potential off-target effects on other POZ-domain proteins cannot be excluded. This product is intended for laboratory research only.
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| References | |
| Additional Infomation |
SPOP-IN-6b dihydrochloride is a research tool and has not been approved for clinical use. It is a valuable probe for investigating the SPOP-CUL3 ubiquitin ligase axis and the role of SPOP mutations in cancer. While SPOP is an oncogenic target in some cancers, it can act as a tumor suppressor in others. This compound helps elucidate these context-dependent functions. It should be stored at -20degC as a powder and is stable for at least one year.
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| Molecular Formula |
C28H34CL2N6O3
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|---|---|
| Related CAS # |
SPOP-IN-6b;2136270-20-5;SPOP-IN-6b hydrochloride;2136270-68-1
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| Appearance |
Light yellow to yellow 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, 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 :~41.67 mg/mL (~72.66 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.63 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 20.8 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. (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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