Neriifolin is a cardiac glycoside that penetrates the central nervous system/CNS and is an inhibitor (blocker/antagonist) of Na+, K+-ATPase. Neriifolin targets beclin 1, inhibits the formation of LC3-related phagosomes, and improves the development of experimental autoimmune encephalomyelitis (EAE). Neriifolin induces cell cycle arrest and apoptosis in human liver cancer HepG2 cells.

Physicochemical Properties

Molecular Formula	C30H46O8
Molecular Weight	534.68
Exact Mass	534.319
CAS #	466-07-9
PubChem CID	441867
Appearance	White to yellow solid powder
Density	1.274g/cm3
Boiling Point	700.079ºC at 760 mmHg
Melting Point	218-225ºC
Flash Point	224.862ºC
Vapour Pressure	0mmHg at 25°C
Index of Refraction	1.581
LogP	3.11
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	4
Heavy Atom Count	38
Complexity	968
Defined Atom Stereocenter Count	13
SMILES	C[C@H]1[C@@H]([C@H]([C@@H]([C@@H](O1)O[C@H]2CC[C@]3([C@@H](C2)CC[C@@H]4[C@@H]3CC[C@]5([C@@]4(CC[C@@H]5C6=CC(=O)OC6)O)C)C)O)OC)O
InChi Key	VPUNMTHWNSJUOG-BAOINKAISA-N
InChi Code	InChI=1S/C30H46O8/c1-16-24(32)26(35-4)25(33)27(37-16)38-19-7-10-28(2)18(14-19)5-6-22-21(28)8-11-29(3)20(9-12-30(22,29)34)17-13-23(31)36-15-17/h13,16,18-22,24-27,32-34H,5-12,14-15H2,1-4H3/t16-,18+,19-,20+,21-,22+,24-,25-,26+,27-,28-,29+,30-/m0/s1
Chemical Name	3-[(3S,5R,8R,9S,10S,13R,14S,17R)-3-[(2R,3S,4R,5S,6S)-3,5-dihydroxy-4-methoxy-6-methyloxan-2-yl]oxy-14-hydroxy-10,13-dimethyl-1,2,3,4,5,6,7,8,9,11,12,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]-2H-furan-5-one
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
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

ln Vitro	In HepG2 cells, nerifolin (0.1μg/mL; 48 hours) causes apoptosis. The viability of HepG2 cells is reduced by nerifolin (0–8 μg/mL; 72 hours). Additionally, nerifolin causes cell cycle arrests in the S and G2/M phases and promotes HepG2 cell death. Neriifolin stimulation of HepG2 cells resulted in upregulated production of the Fas and FasL proteins as well as the activation of caspase-3, -8, and -9[1].
References	[1]. ATG-dependent phagocytosis in dendritic cells drives myelin-specific CD4+ T cell pathogenicity during CNS inflammation. Proc Natl Acad Sci U S A. 2017;114(52):E11228-E11237. [2]. Neriifolin from seeds of Cerbera manghas L. induces cell cycle arrest and apoptosis in human hepatocellular carcinoma HepG2 cells. Fitoterapia. 2011;82(5):735-741.
Additional Infomation	Neriifolin is a cardenolide glycoside that is digitoxigenin in which the hydroxy goup at position 3 has been converted to its (6-deoxy-3-O-methyl-alpha-L-glucopyranoside derivative. Found in the seeds of Cerbera odollamand in Thevetia ahouia and Thevitia neriifolia. It has a role as a cardiotonic drug, a toxin and a neuroprotective agent. It is functionally related to a digitoxigenin. Neriifolin has been reported in Thevetia ahouai, Cerbera odollam, and other organisms with data available.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (187.03 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.68 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 (4.68 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8703 mL	9.3514 mL	18.7028 mL
	5 mM	0.3741 mL	1.8703 mL	3.7406 mL
	10 mM	0.1870 mL	0.9351 mL	1.8703 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.