FTY720

Biological Activity

FTY720 (Fingolimod) is a sphingosine-1-phosphate receptor 1 receptor. FTY720 is phosphorylated by sphingosine kinase, which then acts as a potent agonist at four of the sphingosine-1-phosphate (S1P) receptors (S1P1, S1P3, S1P4, and S1P5). FTY720 (Fingolimod) also mediated antitumor effects in hepatocellular carcinoma (HCC) cells by activating PKC. The three HCC cell lines examined, Hep3B, Huh7, and PLC5, exhibited differential susceptibility to FTY720-mediated suppression of cell viability, with IC50 values of 4.5, 6.3, and 11 μM, respectively.

Product Citations

• 

  Neuropharmacology. 2017 May 1;117:149-157.

  FTY720 (Fingolimod) reverses a-synuclein-induced downregulation of brain-derived neurotrophic factor mRNA in OLN-93 oligodendroglial cells
  FTY720 purchased from AbMole

• 

  Powder Diffraction. 2015;205-210.

  Crystal structure of fingolimod hydrochloride, C19H34ClNO2
  FTY720 purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Neuropharmacology (2017). Figure 6. FTY720 (AbMole BioScience, Kowloon, Hong Kong, China)
	Method	Immunoblots
	Cell Lines	OLN-93 cells
	Concentrations	160 nM
	Incubation Time	12 h
	Results	On immunoblots we noted that 12 h of 160 nM FTY720 treatment of OLN-93 cells produced a significant increase in global acetylation of histone 3 (AcH3)

	Source	Neuropharmacology (2017). Figure 4. FTY720 (AbMole BioScience, Kowloon, Hong Kong, China)
	Method	qPCR
	Cell Lines	OLN-93 cells
	Concentrations	160 nM
	Incubation Time	12 h
	Results	Empty vector transfected cells expressed BDNF (Fig. 4A, lane 1) and 12 h treatments with 160 nM FTY720 significantly increased BDNF expression in all stably transfected aSyn OLN-93 cell lines (Fig. 4A, lanes 3, 5 and 7), visualized on agarose gels showing the BDNF amplicons generated by qPCR.

	Source	Neuropharmacology (2017). Figure 2. FTY720 (AbMole BioScience, Kowloon, Hong Kong, China)
	Method	qPCR
	Cell Lines	OLN-93 cells
	Concentrations	160 nM
	Incubation Time	12 h
	Results	Similar to our earlier assessment in neuronal cells (Vargas-Medrano et al., 2014), we found that using 160 nM FTY720 treatment of OLN-93 cells for 12 h significantly increased BDNF expression as measured using qPCR (Fig. 2, dark gray whisker box).

Protocol

Cell Experiment
Cell lines	MCF-7 cells
Preparation method	In Vitro Proliferation Assay Cells were seeded onto 96- well plates at the densities of 3000 cells/well. After 24 h of pre-cultivation, appropriate concentrations of the samples were added and the cells were cultured for 2 d. The relative growth ratio was determined by WST-1 assay using a cell counting kit (Dojindo Laboratories, Kumamoto, Japan) according to the manufacturer’s instructions. The IC50 value was determined from the dose response curve. At least three independent experiments were performed. The proliferation assay that tested the effects of DMS was performed using identical methods. Compounds with or without DMS were added to the wells and the plates were incubated for 78 h at 37 °C in a humidified atmosphere containing 5% CO2 before the WST-1 assay was conducted.
Concentrations	0~100µM
Incubation time	2 days

Animal Experiment
Animal models	Six-weekold female NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice with Mino cells
Formulation	not mentioned
Dosages	10 mg/kg daily for 10 days
Administration	intraperitoneal injections

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Chemical Information

References

Fingolimod (FTY720) inhibits neuroinflammation and attenuates spontaneous convulsions in lithium-pilocarpine induced status epilepticus in rat model.
Gao et al. Pharmacol Biochem Behav. 2012 Aug 31. PMID: 22960129.

Fingolimod in the treatment algorithm of relapsing remitting multiple sclerosis: a statement of the Central and East European (CEE) MS Expert Group.
Fazekas F. Wien Med Wochenschr. 2012 Aug;162(15-16):354-366. PMID: 22895849.

Fingolimod, a sphingosine-1 phosphate receptor modulator, increases BDNF levels and improves symptoms of a mouse model of Rett syndrome.
Deogracias et al. Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):14230-5. PMID: 22891354.

Development of oral agent in the treatment of multiple sclerosis: how the first available oral therapy, fingolimod will change therapeutic paradigm approach.
Gasperini et al. Drug Des Devel Ther. 2012;6:175-86. PMID: 22888218.