| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
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| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
IC50: 2 nM (SPHK1); 26.7 nM (Sphingosine 1-phosphate (S1P))[1] Ki: 3.6 nM (SPHK1)[1]
Sphingosine kinase 1 (SphK1). PF-543 HCl is a potent and selective inhibitor of SphK1 with an IC50 of 2.0 nM and a Ki of 3.6 nM. It is sphingosine-competitive and is more than 100-fold selective for SphK1 over the SphK2 isoform. |
|---|---|
| ln Vitro |
At nM concentrations, PF-543 (10-1000 nM; 24 hours; PASM cells) treatment eliminates SK1 expression[2]. Treatment of PASM cells with PF-543 (0.1-10 μM) for 24 hours causes caspase-3/7 activity[2]. With an IC50 of 1.0 nM, PF-543 suppresses the production of C17-S1P in 1483 cells[1]. In 1483 cells, PF-543 inhibits SphK1, leading to a dose-dependent decrease in the intracellular concentration of S1P (EC50 = 8.4 nM) and an increase in the intracellular concentration of sphingosine. Following a one-hour treatment with 200 nM PF-543, the amount of endogenous S1P in 1483 cells is reduced tenfold, leading to a corresponding increase in the level of sphingosine[1].
PF-543 HCl is a potent SphK1 inhibitor with an IC50 of 2.0 nM and a Ki of 3.6 nM. It is sphingosine-competitive and more than 100-fold selective for SphK1 over SphK2. PF-543 (10-1000 nM; 24 hours) shows antitumor activity. |
| ln Vivo |
Treatment with PF-543 (1 mg/kg; intraperitoneal injection; every other day; for 21 days; female C57BL/6 J mice) decreases right ventricular hypertrophy but has no effect on vascular remodelling. Antioxidant nuclear factor Nrf-2 expression is upregulated and p53 expression is downregulated as a result of the protection[2]. Blood samples are given to mice i.p. at 10 mg/kg or 30 mg/kg of PF-543 for a duration of 24 hours. The T1/2 is 1.2 hours. Mice given 10 mg/kg PF-543 for 24 hours experience a reduction in SK1 expression in their pulmonary vessels[2].
PF-543 (1 mg/kg; intraperitoneal injection; every second day; for 21 days) treatment has no effect on vascular remodelling but reduces right ventricular hypertrophy in mice. The protection involves a reduction in the expression of p53. PF-543 HCl has potential therapeutic applications in the treatment of cancer. |
| Enzyme Assay |
SphK1 enzyme activity assays are performed using recombinant human SphK1 enzyme. The enzyme is incubated with the substrate sphingosine and [γ-32P]ATP in assay buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.1% Triton X-100). The reaction is incubated at 37°C for 30-60 minutes. The product sphingosine-1-phosphate (S1P) is extracted and quantified by scintillation counting or by LC-MS/MS. PF-543 HCl is serially diluted and added to the reaction mixture. IC50 and Ki values are determined by non-linear regression analysis. Each concentration is tested in duplicate.
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| Cell Assay |
Western Blot Analysis[2]
Cell Types: Human pulmonary arterial smooth muscle (PASM) cells Tested Concentrations: 10 nM, 100 nM, 1000 nM Incubation Duration: 24 hrs (hours) Experimental Results: Abolished SK1 expression at nM concentrations. Apoptosis Analysis[2] Cell Types: Human pulmonary arterial smooth muscle (PASM) cells Tested Concentrations: 0.1 μM, 1 μM, 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Induced caspase-3/7 activity in cultured human pulmonary smooth muscle cells. Cellular SphK1 inhibition is evaluated in cancer cell lines. Cells are cultured in appropriate media at 37°C with 5% CO2 and treated with PF-543 HCl at various concentrations (10-1000 nM) for 24 hours. SphK1 activity in cell lysates is measured using enzymatic assays. S1P levels in cells and culture supernatants are quantified by LC-MS/MS. Cell viability and proliferation are assessed using MTT or CellTiter-Glo assays. Apoptosis is evaluated by Annexin V/PI staining. Each experiment includes known SphK1 inhibitors as positive controls and vehicle controls. |
| Animal Protocol |
Animal/Disease Models: Female C57BL/6 J mice (7-12 week-old) with hypoxic-induced pulmonary arterial hypertension[2]
Doses: 1 mg/kg Route of Administration: intraperitoneal (ip)injection; every second day; for 21 days Experimental Results: decreased right ventricular hypertrophy. The protection involves a reduction in the expression of p53 (that promotes cardiomyocyte death) and an increase in the expression of anti-oxidant nuclear factor Nrf-2. In vivo efficacy is evaluated in mouse models. PF-543 is administered intraperitoneally at 1 mg/kg every second day for 21 days. Vascular remodelling and right ventricular hypertrophy are assessed. p53 expression is measured by Western blotting. For antitumor studies, xenograft models are used, and tumor growth is monitored. Sample sizes typically range from 6-10 animals per group. |
| ADME/Pharmacokinetics |
Molecular Weight: 502.07. Formula: C22H31NO4S·HCl. CAS No.: 1706522-79-3. Purity: >98% by HPLC. Solubility: Soluble in DMSO, not in water. Target: SphK1. For research use only.
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| Toxicity/Toxicokinetics |
No comprehensive toxicology data are publicly available. As a SphK1 inhibitor, potential toxicities may include effects on sphingolipid metabolism and cell signaling. Standard toxicity studies would include acute and subchronic toxicity in rodents, genotoxicity screening, and evaluation of effects on the cardiovascular system and cell proliferation. No clinical trials have been reported for this compound. The compound is intended for research use only.
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| References |
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| Additional Infomation |
PF-543 HCl is also known as PF-543. It is the most potent inhibitor of SphK1 described to date, with an IC50 of 2.0 nM and a Ki of 3.6 nM. It is useful for dissecting specific roles of SphK1-driven S1P signalling. No clinical trials or regulatory approvals have been reported. The compound is for research use only.
|
| Molecular Formula |
C27H32CLNO4S
|
|---|---|
| Molecular Weight |
502.065285682678
|
| Exact Mass |
501.174
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| CAS # |
1706522-79-3
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| Related CAS # |
PF-543;1415562-82-1;PF-543 Citrate;1415562-83-2
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| PubChem CID |
121230770
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
9
|
| Heavy Atom Count |
34
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| Complexity |
679
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| Defined Atom Stereocenter Count |
1
|
| SMILES |
CC1=CC(=CC(=C1)OCC2=CC=C(C=C2)CN3CCC[C@@H]3CO)CS(=O)(=O)C4=CC=CC=C4.Cl
|
| InChi Key |
WNKWAZFYPZMDGJ-VQIWEWKSSA-N
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| InChi Code |
InChI=1S/C27H31NO4S.ClH/c1-21-14-24(20-33(30,31)27-7-3-2-4-8-27)16-26(15-21)32-19-23-11-9-22(10-12-23)17-28-13-5-6-25(28)18-29;/h2-4,7-12,14-16,25,29H,5-6,13,17-20H2,1H3;1H/t25-;/m1./s1
|
| Chemical Name |
[(2R)-1-[[4-[[3-(benzenesulfonylmethyl)-5-methylphenoxy]methyl]phenyl]methyl]pyrrolidin-2-yl]methanol;hydrochloride
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| Synonyms |
PF543 PF-543 HCl PF 543 PF543
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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 | 1.9918 mL | 9.9588 mL | 19.9175 mL | |
| 5 mM | 0.3984 mL | 1.9918 mL | 3.9835 mL | |
| 10 mM | 0.1992 mL | 0.9959 mL | 1.9918 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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