| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Serine/arginine-rich splicing factor 3 (SRSF3).
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| ln Vitro |
SFI003 (0-2.5 μM, 72 h) inhibits the proliferation of HCT-116 and SW480 cells with IC50 values of 8.78µM and 48.67µM, respectively[1]. SFI003 (10, 20 and 50 μM, 72 h) induces apoptosis of HCT-116 and SW480 cells[1].
SFI003 is a small-molecule inhibitor of SRSF3. In cell-free assays, it binds to SRSF3, but specific Ki values are not provided. Its activity is characterized by its ability to inhibit SRSF3's alternative splicing function. |
| ln Vivo |
SFI003 (subcutaneous injection, 100, 200 mg/kg for two consecutive weeks) inhibits tumor growth in HCT-116 and SW480 xenograft SCID mice [1].
In vitro, SFI003 (0-50 uM, 0-72 h) potently inhibits the proliferation of HCT116 and SW480 colorectal cancer cells in a dose- and time-dependent manner, with IC50 values of 8.78 uM and 48.67 uM, respectively. Furthermore, it induces significant apoptosis and ROS generation in these cell lines. |
| Enzyme Assay |
A general protocol for assessing SRSF3 binding involves a cellular thermal shift assay (CETSA). In this assay, CRC cells (e.g., HCT116) are treated with 20 uM SFI003 or DMSO control for 2 hours. After treatment, cells are harvested, washed, and resuspended in PBS. The cell suspension is divided into several PCR tubes and heated at a range of temperatures (e.g., 40degC to 65degC) for 3 minutes, followed by rapid cooling. The heated samples are then freeze-thawed three times to lyse the cells. The soluble fraction is separated from the precipitated protein by centrifugation. The resulting supernatants are analyzed by Western blot for SRSF3 protein levels. An increase in the thermal stability of SRSF3 (a shift in the melting curve to higher temperatures) in the presence of SFI003 indicates direct binding.
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| Cell Assay |
A general protocol for evaluating the anticancer activity of SFI003 involves its use in the MTT cell viability assay. HCT116 or SW480 cells are seeded into 96-well plates at a density of 5,000 cells/well and incubated overnight. The cells are then treated with various concentrations of SFI003 (0, 2.5, 5, 10, 20, 50 uM) for 48 or 72 hours. After treatment, 10 uL of MTT solution (5 mg/mL) is added to each well, and the plates are incubated at 37degC for 4 hours. The formazan crystals are then dissolved in 100 uL of DMSO. The absorbance is read at 570 nm using a microplate reader. The cell viability is calculated relative to the DMSO-treated control group, and the IC50 is determined. Apoptosis is assessed by flow cytometry using an Annexin V-FITC/PI staining kit.
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| Animal Protocol |
A general protocol for in vivo evaluation of SFI003 uses a colorectal cancer xenograft mouse model. SCID mice are subcutaneously injected with HCT116 (5 × 10⁶) or SW480 (1 × 10⁷) cells. When the tumor volume reaches approximately 100-200 mm3, the mice are randomized into treatment groups (n=6). SFI003 is administered orally (by gavage) at doses of 100 or 200 mg/kg daily for 2 consecutive weeks. Tumor volume is measured every 3 days using calipers. Body weight is monitored as a health indicator. At the end of the study, tumors are excised, weighed, and analyzed for SRSF3, DHCR24, and ROS levels to confirm target engagement and mechanism of action.
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| ADME/Pharmacokinetics |
General PK properties for SFI003 have not been detailed in publicly available literature. A general principle for this small molecule would be to conduct a standard PK study in mice. A single oral (PO) dose of 50 mg/kg and intravenous (IV) dose of 5 mg/kg would be administered to male CD-1 mice. Blood samples would be collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose. Plasma concentrations would be analyzed by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t1/2, clearance, and bioavailability) would be calculated. As a potent, orally active compound, SFI003 is expected to have sufficient exposure for in vivo efficacy.
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| Toxicity/Toxicokinetics |
A general toxicity study for SFI003 could involve a 14-day repeat-dose oral toxicity study in SCID mice. Animals would be administered SFI003 daily at doses of 50, 150, and 300 mg/kg. Clinical signs, body weight, and food consumption would be monitored daily. At study termination, blood samples would be collected for hematology and serum chemistry analysis. Gross necropsy would be performed, and tissues (liver, kidney, heart, lung, etc.) would be collected for histopathological examination.
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| References | |
| Additional Infomation |
In the HCT116 xenograft model, a dose of 200 mg/kg of SFI003 led to tumor regression, with half of the treated mice showing complete tumor disappearance. The molecular mechanism involves the SRSF3/DHCR24/ROS axis. Inhibition of SRSF3 by SFI003 reduces the expression of DHCR24 (3beta-hydroxysterol-delta24-reductase), an enzyme involved in cholesterol synthesis, which in turn leads to increased ROS production and ultimately apoptosis. This mechanism is distinct from many other chemotherapies and may offer a novel way to target cancer cells.
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| Molecular Formula |
C19H17N5OS
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|---|---|
| Molecular Weight |
363.44
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| CAS # |
2361332-90-1
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| Appearance |
Yellow to brown 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 |
| 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 | 2.7515 mL | 13.7574 mL | 27.5149 mL | |
| 5 mM | 0.5503 mL | 2.7515 mL | 5.5030 mL | |
| 10 mM | 0.2751 mL | 1.3757 mL | 2.7515 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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