Sphingosine 1-phosphate (S1P) is an extracellular ligand that regulates endothelial cell function and angiogenesis, promotes survival and migration of cancer cells, and acts as a key regulator of immune cell trafficking. S1P is formed primarily by the action of sphingosine kinases (SphKs). PF 543 is a potent, cell-permeant inhibitor of SphK1 with an IC50 value of 2.0 ± 0.6nM in an in vitro enzyme assay and is able to inhibit the enzyme >95% at 20 nM. PF-543 was also able to inhibit C17-S1P formation in 1483 cells with an IC50 value of 1.0 ± 0.3nM. PF-543 showed 132-fold selectivity for SphK1 over SphK2 (IC50 = 356nM). The inhibition of SphK1 by PF-543 led to a concentration-dependent depletion of the intracellular level of S1P with an EC50 value of 8.4nM. The presence of 1μM PF-543 for 7 days in the culture medium resulted in a 10-fold decrease in S1P and a 2-fold increase in sphingosine levels in 1483 cells. PF543 also potently inhibited C17-S1P formation in human whole blood with an IC50 value of 26.7 nM[1]. PF-543 treatment (0.93mg/kg body weight per day) for 28 days significantly decreased erythrocyte Sphk1 activity and reduced erythrocyte and plasma S1P levels in SCD Tg mice[2].
作用机制
PF 543 structurally resembles sphingosine and potentially could act as SphK1 substrate. It inhibits S1P binding to human recombinant SphK1 with a Ki value of 3.6nM[1].
PF543 pretreatment significantly attenuated neuronal apoptosis induced by M1 microglia.
J Neuroinflammation. 2021 Feb 18;18(1):50.
Huh7 HCC cells
5 µM
16 hours
To evaluate the effects of PF-543 on Huh7 HCC cells, results showed that PF-543 significantly reduced S1P levels but did not affect cell viability, cell death, or clonogenicity
J Transl Med. 2024 Jan 10;22(1):43.
Human umbilical vein endothelial cells (HUVECs)
5 µM
16 hours
To evaluate the effects of PF-543 on endothelial cell angiogenesis, results showed that PF-543 significantly inhibited tube formation, migration, and sprouting but did not affect cell viability or cell death
J Transl Med. 2024 Jan 10;22(1):43.
Human head and neck SCC cell lines
1, 5, 10, 25, or 50 µM
24, 48, and 72 hours
To assess cell viability and types of cell death induced by PF-543.
Cell Death Discov. 2017 Aug 14;3:17047.
Sickle cells
up to 50 µM
PF-543 inhibited SPHK1 activity and reduced sickle cell formation under hypoxic conditions without causing hemolysis
J Clin Invest. 2014 Jun;124(6):2750-61.
PF-543 HCl 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
SCD Tg mice
Sickle cell disease model
Subcutaneous injection
0.93 mg/kg/day
Once daily for 28 days
PF-543 treatment significantly reduced SPHK1 activity, S1P levels, and the percentage of sickle cells in SCD mice, and also reduced hemolysis and inflammation
J Clin Invest. 2014 Jun;124(6):2750-61.
Sprague-Dawley rats
Spinal cord injury model
Intraperitoneal injection
10 mg/kg
Daily until sacrifice
PF543 attenuated inflammation and neuronal damage after spinal cord injury, leading to improved functional recovery.
J Neuroinflammation. 2021 Feb 18;18(1):50.
Mice
Ang II-infused hypertensive mice
Injection
10 mg/kg
Every 2 days
PF543 mitigated cardiac hypertrophy and improved endothelial function without affecting blood pressure.
Hypertension. 2020 Feb;75(2):383-392
SCD Tg mice
Diethylnitrosamine (DEN)-induced primary liver cancer model
Intraperitoneal injection
25 mg/kg
Every other day for 12 weeks
To evaluate the inhibitory effects of PF-543 on DEN-induced primary liver cancer, results showed that PF-543 significantly inhibited SphK1 activity, reduced tumor number and size, and decreased tumor angiogenesis
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