PF-543 Citrate (PF-543; PF 543), the citrate salt of PF-543, is a novel cell-permeable and sphingosine-competitive SphK1 inhibitor with potential anticancer activity. It inhibits inhibits SphK1 with IC50s and Ki of 2.0 nM and 3.6 nM, and exhibits >100-fold selectivity for SphK1 over the SphK2 isoform.
Physicochemical Properties
| Molecular Formula | C33H39NO11S |
| Molecular Weight | 657.727869272232 |
| Exact Mass | 657.224 |
| CAS # | 1415562-83-2 |
| Related CAS # | PF-543;1415562-82-1;PF-543 hydrochloride;1706522-79-3 |
| PubChem CID | 71576669 |
| Appearance | White to yellow solid powder |
| LogP | 4.274 |
| Hydrogen Bond Donor Count | 5 |
| Hydrogen Bond Acceptor Count | 12 |
| Rotatable Bond Count | 14 |
| Heavy Atom Count | 46 |
| Complexity | 906 |
| 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.C(C(=O)O)C(CC(=O)O)(C(=O)O)O |
| InChi Key | PWXXWUWKNPXSGW-VQIWEWKSSA-N |
| InChi Code | InChI=1S/C27H31NO4S.C6H8O7/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;7-3(8)1-6(13,5(11)12)2-4(9)10/h2-4,7-12,14-16,25,29H,5-6,13,17-20H2,1H3;13H,1-2H2,(H,7,8)(H,9,10)(H,11,12)/t25-;/m1./s1 |
| Chemical Name | [(2R)-1-[[4-[[3-(benzenesulfonylmethyl)-5-methylphenoxy]methyl]phenyl]methyl]pyrrolidin-2-yl]methanol;2-hydroxypropane-1,2,3-tricarboxylic acid |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
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| ln Vitro | SK1 expression is eliminated at nM concentrations when PASM cells are treated with PF-543 Citrate (10-1000 nM) for 24 hours [2]. Caspases-3/7 are activated when treated with PF-543 Citrate (0.1–10 μM) for 24 hours in PASM cells [2]. With an IC 50 of 1.0 nM, PF-543 Citrate suppresses the production of C17-S1P in 1483 cells[1]. When PF-543 Citrate inhibited SphK1, intracellular S1P levels were depleted in a dose-dependent manner with an EC50 value of 8.4 nM, while intracellular sphingosine levels increased in 1483 cells concurrently. 1483 cells' endogenous S1P levels were tenfold decreased and sphingosine levels increased following an hour-long treatment with 200 nM PF-543 Citrate [1]. | |
| ln Vivo | While PF-543 Citrate (1 mg/kg; i.p.; every other day; for 21 days; female C57BL/6 J mice) decreased right ventricular hypertrophy, it had no effect on vascular remodeling. Protection entails elevated expression of the antioxidant nuclear protein Nrf-2 and decreased expression of p53 [2]. T1/2 in blood samples was 1.2 hours after mice were first given PF-543 Citrate at doses of 10 mg/kg or 30 mg/kg for a duration of 24 hours. Mice given 10 mg/kg PF-543 Citrate for a full day showed a reduction in SK1 expression in their pulmonary arteries [2]. | |
| 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. |
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| 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. |
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| References |
[1]. Modulation of cellular S1P levels with a novel, potent and specific inhibitor of sphingosine kinase-1. Biochem J. 2012 May 15;444(1):79-88. [2]. Effect of the sphingosine kinase 1 selective inhibitor, PF-543 on arterial and cardiac remodelling in a hypoxic model of pulmonary arterial hypertension. Cell Signal. 2016 Aug;28(8):946-55. [3]. Induction of autophagy by sphingosine kinase 1 inhibitor PF-543 in head and neck squamous cell carcinoma cells. Cell Death Discov. 2017 Aug 14;3:17047. |
Solubility Data
| Solubility (In Vitro) |
DMSO : ≥ 100 mg/mL (~152.04 mM) H2O : ~50 mg/mL (~76.02 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.80 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 25.0 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: ≥ 2.5 mg/mL (3.80 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (3.80 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 100 mg/mL (152.04 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5204 mL | 7.6019 mL | 15.2038 mL | |
| 5 mM | 0.3041 mL | 1.5204 mL | 3.0408 mL | |
| 10 mM | 0.1520 mL | 0.7602 mL | 1.5204 mL |