| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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Purity: ≥98%
Clofilium tosylate, the tosylate salt of clofilium, is a novel and potent potassium/K+ channel blocker with the potential to be used as a cardiac depressant and anti-arrhythmic agent. It induces apoptosis of human promyelocytic leukemia (HL-60) cells via Bcl-2-insensitive activation of caspase-3.
| Targets |
Potassium channel blocker . The apoptotic effect is reported to be independent of its potassium channel blocking activity at the concentrations used (2.5–10 μM). [1]
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| ln Vitro |
Clofilium (0–20 μM) treatment of HL-60 cells for 24, 48, and 72 hours inhibited viability and proliferation in a concentration- and time-dependent manner. When HL-60 cells are exposed to 2.5 μM to 10 μM Clofilium (10 μM, 12 h), the protein preparation of inactive procaspase-3, p34 into its active form, p17, is induced. After two hours, the cells are exposed to 10 mM Clofilium. Cell viability is dramatically decreased by substrate PARP [1].
Clofilium inhibits viability and proliferation of human promyelocytic leukemia (HL-60) cells in a time- and concentration-dependent manner. The IC50 values for cell viability reduction were approximately 6.3 μM (24 h), 3.4 μM (48 h), and 2.4 μM (72 h). [1] Clofilium induces apoptosis in HL-60 cells, as confirmed by nuclear DAPI staining (showing fragmented apoptotic bodies), transmission electron microscopy (revealing marginated chromatin condensation and nuclear fragmentation), and DNA laddering on agarose gel electrophoresis. [1] Flow cytometry with FITC-Annexin V/PI staining showed that the apoptotic cell population (Annexin V⁺/PI⁻) increased from <2% (0 h) to 20% (4 h) and 29% (16 h) after treatment with 10 μM clofilium. [1] Caspase-3 activity increased approximately 10-fold at 2–3 h after exposure to 10 μM clofilium, as measured by fluorometric immunosorbent enzyme assay (FIENA). [1] Immunoblot analysis demonstrated proteolytic cleavage of procaspase-3 (p34 to p17) and subsequent cleavage of PARP, while expression levels of Bcl-2 and Bax proteins remained unchanged. [1] |
| Enzyme Assay |
Caspase-3 activity was measured using a fluorometric immunosorbent enzyme assay (FIENA). Cells were treated with 10 μM clofilium for various time intervals, harvested, and lysed. The lysates were incubated in anti-caspase-3 coated microplate wells, followed by addition of substrate solution. Activity was determined fluorometrically. [1]
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: HL-60 cells Tested Concentrations: 0-20 μM Incubation Duration: 24, 48, and 72 hrs (hours) Experimental Results: Inhibition of HL-60 cells, the IC50 at 24 hrs (hours) is 6.3 μM, and the IC50 at 48 hrs (hours) is 3.4 μM, IC50 at 72 hrs (hours) is 2.4 μM. Western Blot Analysis[1] Cell Types: HL-60 Cell Tested Concentrations: 10 μM Incubation Duration: 12 hrs (hours) Experimental Results: Proteolytic cleavage of caspase-3 and its substrate PARP was induced, while Bcl-2 and Bax proteins were unchanged. Cell viability was assessed using the MTT dye reduction method. HL-60 cells were treated with clofilium (0–20 μM) for 24, 48, and 72 h. [1] Nuclear morphology was examined by DAPI staining. Cells were fixed, stained with DAPI, and observed under fluorescence microscopy. [1] Transmission electron microscopy was performed. Cells were fixed in glutaraldehyde, post-fixed in osmium tetroxide, dehydrated, embedded, sectioned, stained with uranyl acetate and lead acetate, and examined under an electron microscope. [1] DNA fragmentation analysis was conducted. DNA was extracted using phenol-chloroform method, electrophoresed on agarose gel, stained with ethidium bromide, and visualized under UV light. [1] Apoptosis was quantified by flow cytometry using FITC-Annexin V and propidium iodide (PI) double staining. Cells were incubated with FITC-Annexin V and PI, then analyzed by flow cytometry. [1] Immunoblot analysis was performed to detect caspase-3, PARP, Bcl-2, and Bax proteins. Cells were lysed, proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, blocked, incubated with primary and secondary antibodies, and detected by chemiluminescence. [1] |
| References | |
| Additional Infomation |
Clofilium is a potassium channel blocker that has been used as an antiarrhythmic drug. This study suggests that it has the potential to be used as a chemotherapy drug or cell inhibitor for human leukemia and lymphoma due to its ability to induce apoptosis via a Bcl-2-insensitive, caspase-3-dependent pathway. [1]
The pro-apoptotic effect of clofilium at concentrations of 2.5–10 μM is independent of its potassium channel blocking activity, which typically requires higher concentrations. [1] The proposed mechanism involves plasma membrane receptor-mediated pathways (e.g., CD95/TNFR-1) that are sensitive to Crm A but insensitive to Bcl-2, ultimately leading to caspase-3 activation. [1] |
| Molecular Formula |
C28H44CLNO3S
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|---|---|
| Molecular Weight |
510.1719
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| Exact Mass |
509.273
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| CAS # |
92953-10-1
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| Related CAS # |
68379-02-2;92953-10-1 (Tosylate);68379-03-3 (Phosphate);
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| PubChem CID |
175533
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| Appearance |
White to off-white solid powder
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| LogP |
8.469
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
34
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| Complexity |
460
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
MOQZYUUHIWPDQC-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C21H37ClN.C7H8O3S/c1-4-7-8-9-11-18-23(5-2,6-3)19-12-10-13-20-14-16-21(22)17-15-20;1-6-2-4-7(5-3-6)11(8,9)10/h14-17H,4-13,18-19H2,1-3H3;2-5H,1H3,(H,8,9,10)/q+1;/p-1
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| Chemical Name |
N-(4-(4-chlorophenyl)butyl)-N,N-diethylheptan-1-aminium 4-methylbenzenesulfonate
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| Synonyms |
Clofilium Tosylate
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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 (~196.01 mM)
H2O : ~1.59 mg/mL (~3.12 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.08 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 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.08 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 20.8 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.08 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 2 mg/mL (3.92 mM) in 8g/L NaCl solution (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9601 mL | 9.8007 mL | 19.6013 mL | |
| 5 mM | 0.3920 mL | 1.9601 mL | 3.9203 mL | |
| 10 mM | 0.1960 mL | 0.9801 mL | 1.9601 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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