PF-543, 1415562-82-1,IC-03532168
Product Introduction
PF-543 is a potent, selective, and reversible small-molecule inhibitor of sphingosine kinase 1 (SphK1) with an IC₅₀ of approximately 2 nM and a Ki of 3.6 nM. It exhibits greater than 100-fold selectivity for SphK1 over the SphK2 isoform, making it one of the most selective SphK1 inhibitors available. Sphingosine kinases catalyze the phosphorylation of sphingosine to sphingosine-1-phosphate (S1P), a bioactive lipid mediator that signals through five G protein-coupled S1P receptors (S1PR1-5) to regulate cell proliferation, survival, migration, angiogenesis, and immune cell trafficking. By specifically inhibiting SphK1, PF-543 reduces intracellular S1P levels and induces apoptosis in SphK1-dependent cancer cells. PF-543 has been extensively utilized in cancer biology and lipid signaling research to dissect SphK1-specific functions in tumor growth, chemoresistance, and inflammatory signaling, and to evaluate SphK1 as a therapeutic target.
Product Features
1. Extremely Potent SphK1 Inhibitor: Inhibits SphK1 with an IC₅₀ of ~2 nM and Ki of 3.6 nM, representing one of the most potent SphK1 inhibitors available.
2. Exceptional Isoform Selectivity: Exhibits greater than 100-fold selectivity for SphK1 over SphK2, enabling specific interrogation of SphK1-dependent signaling.
3. Cell-Permeable: Readily crosses cell membranes to inhibit intracellular SphK1 and reduce cellular S1P levels.
4. High Purity: Supplied at ¡Ý98% purity for reproducible pharmacological studies.
5. Extensively Validated: Widely cited in sphingolipid signaling research for studying SphK1 biology, S1P-mediated pathways, and cancer cell apoptosis.
Specifications
Size: Available in 5 mg and 10 mg
CAS Number: 1415562-82-1
Purity: ¡Ý98%
Molecular Weight: 417.56
Molecular Formula: C23H27NO4S
Solubility: DMSO: ¡Ý20 mg/mL; limited solubility in aqueous buffers
Storage and Stability
Storage Conditions: Store at -20¡ãC, protected from light and moisture.
Shelf Life: The product is stable for 24 months from the date of manufacture when stored as directed. For reconstituted solutions, aliquot and store at -20¡ãC. Avoid repeated freeze-thaw cycles.
Protocol (For Reference Only)
Important: PF-543 is light-sensitive. Prepare stock solutions in anhydrous DMSO and protect from light during handling and storage.
1. Stock Solution Preparation: Dissolve PF-543 in anhydrous DMSO to prepare a concentrated stock solution (e.g., 10-50 mM). Aliquot into single-use vials and store at -20¡ãC protected from light.
2. In Vitro Working Solution Preparation: Dilute DMSO stock solution into pre-warmed culture medium to achieve working concentrations typically ranging from 0.1-10 µM. Limit final DMSO concentration in culture medium to ¡Ü0.1% to avoid solvent-related cytotoxicity.
3. SphK1 Inhibition Assay: For SphK1 inhibition, treat cells with PF-543 at 0.1-5 µM for 1-6 hours. Assess SphK1 activity by measuring intracellular S1P levels by LC-MS/MS or ELISA, or evaluate sphingosine accumulation. For complete and sustained SphK1 inhibition, 1-5 µM is recommended.
4. Apoptosis and Cell Viability Assays: Treat SphK1-dependent cancer cells with PF-543 at 1-10 µM for 24-72 hours. Assess cell viability by MTT assay and apoptosis by annexin V/PI flow cytometry or caspase-3/7 activation assays. Include SphK1-independent cell lines as negative controls.
5. S1P Signaling Analysis: Following PF-543 treatment, evaluate downstream S1P receptor signaling by assessing ERK and Akt phosphorylation by Western blot, or measure S1P secretion into culture medium by LC-MS/MS or ELISA.
Precautions
1. Protect from light during handling and storage; PF-543 is photodegradable.
2. Use appropriate vehicle controls in all experiments; DMSO concentration in culture medium should not exceed 0.1%.
3. PF-543 exhibits differential potency across cell types depending on SphK1 expression levels and dependence on SphK1-driven S1P production.
4. For in vivo studies, PF-543 has been administered in rodent models; optimize formulation and dosing based on literature references for specific applications.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the recommended concentration for SphK1 inhibition in vitro?
A1: PF-543 at 0.1-5 µM effectively inhibits SphK1 in most cell types. For complete SphK1 inhibition, 1-5 µM is recommended. The IC₅₀ for SphK1 is approximately 2 nM in cell-free assays, but higher concentrations are required in cellular assays due to intracellular protein binding and SphK1 abundance.
Q2: Does PF-543 inhibit SphK2?
A2: PF-543 exhibits greater than 100-fold selectivity for SphK1 over SphK2. At concentrations up to 10 µM, minimal SphK2 inhibition is observed. For dual SphK1/SphK2 inhibition, alternative inhibitors should be used.
Q3: What is the effect of PF-543 on cellular S1P levels?
A3: PF-543 treatment reduces intracellular S1P levels by inhibiting SphK1-catalyzed S1P synthesis. The extent and kinetics of S1P reduction depend on cell type, SphK1 expression, and the contribution of SphK2 to total S1P production.
Q4: Can PF-543 be used in vivo?
A4: PF-543 has been administered in rodent models via intraperitoneal injection. Due to its pharmacokinetic properties, repeated dosing may be required to maintain sustained SphK1 inhibition. Optimize dosing and formulation based on published literature for specific disease models.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of conditions using a small-scale pilot experiment is strongly recommended before processing valuable samples.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-03532168
Product Name: PF-543
Size: 5 mg / 10 mg
Price:
5 mg: CNY ¥950.00 / USD $95.00 / EUR €114.00 / JPY ¥17100.00
10 mg: CNY ¥1650.00 / USD $165.00 / EUR €198.00 / JPY ¥29700.00
2023 Version
