FTY720 hydrochloride

Biological Activity

Fingolimod (FTY720) HCl is a 1-phospho-neuraminol S1P receptor antagonist with an IC50 of 0.33 nM. , FTY720 can dephosphorylate AMPK by activating PP2A and reduce the expression level of phosphorylated eEF2K, ultimately leading to ADCD and iron death in human multiple myeloma cells.

Product Citations

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  Pulm Circ. 2020 Feb 10;10(1).

  Sphingosine-1-phosphate Receptor-Independent Lung Endothelial Cell Barrier Disruption Induced by FTY720 Regioisomers
  FTY720 hydrochloride purchased from AbMole

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  J Nat Prod. 2020 May 20.

  Anti-inflammatory Efficacy of Combined Natural Alkaloid Berberine and S1PR Modulator Fingolimod at Low Doses in Ulcerative Colitis Preclinical Models
  FTY720 hydrochloride purchased from AbMole

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  Neuropharmacology. 2019 Nov 1;158:107701.

  FTY720-Mitoxy reduces toxicity associated with MSA-like α-synuclein and oxidative stress by increasing trophic factor expression and myelin protein in OLN-93 oligodendroglia cell cultures.
  FTY720 hydrochloride purchased from AbMole

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  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 hydrochloride purchased from AbMole

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  Powder Diffraction. 2015;205-210.

  Crystal structure of fingolimod hydrochloride, C19H34ClNO2
  FTY720 hydrochloride 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 Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 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 Compound A of 10 mg/kg.

Chemical Information

References

[1] Cipriani R, et al. J Neuroinflammation. FTY720 attenuates excitotoxicity and neuroinflammation.

[2] Gao et al. Pharmacol Biochem Behav. Fingolimod (FTY720) inhibits neuroinflammation and attenuates spontaneous convulsions in lithium-pilocarpine induced status epilepticus in rat model.

[3] Fazekas F. Wien Med Wochenschr. Fingolimod in the treatment algorithm of relapsing remitting multiple sclerosis: a statement of the Central and East European (CEE) MS Expert Group.

[4] Deogracias et al. Proc Natl Acad Sci U S A. Fingolimod, a sphingosine-1 phosphate receptor modulator, increases BDNF levels and improves symptoms of a mouse model of Rett syndrome.

[5] Gasperini et al. Drug Des Devel Ther. Development of oral agent in the treatment of multiple sclerosis: how the first available oral therapy, fingolimod will change therapeutic paradigm approach.