MNS (3,4-Methylenedioxy-β-nitrostyrene, MDBN) 1485-00-3

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MNS (3,4-methyl-enedioxy-β-nitrostyrene) completely inhibits 2 μM
U46619-(a thromboxane A2 mimic), 5 μM ADP-, 100 μM arachidonic
acid-(AA), 10 μg/ml collagen-, and 0.1 U/ml thrombin-induced platelet
aggregation in a concentration-dependent manner with IC50 of 2.1 μM, 4.1
μM, 5.8 μM, 7.0 μM, and 12.7 μM, respectively. MNS inhibits platelet
aggregation caused by either the calcium ionophore A23187 (1 μM) or the
protein kinase C (PKC) activator PDBu (200 nM) with IC50 of 25.9 μM and
4.8 μM, respectively. MNS (20 μM) decreases dthrombin-induced P-selectin
expression on platelets to levels comparable to those observed in
PGE1-treated platelets.


MNS (20 μM) markedly inhibits thrombin-but not
PDBu-induced MARCKS phosphorylation in platelets. MNS (20 μM) markedly
inhibits protein tyrosine phosphorylation at either 0.5 min or 3 min
after thrombin or collagen stimulation in platelets. MNS
stimulates UbG76V-GFP and ODD-Luc degradation with IC50 of 1.6 μM and
5.9 μM, respectively. MNS inhibits MG132-induced accumulation of the
reporter with IC50 of 2.1 μM. MNS inhibits Gram-positive
(Staphylococcus aureus and Enterococcus faecalis) and Gram-negative
(Escherichia coli and Pseudomonas aeruginosa) bacteria with minimum
inhibitory concentrations (MICs) of 128 mg/L.


MNS is much
more potent than genistein in inhibiting platelet aggregation and
protein tyrosine phosphorylation. MNS
(3,4-Methylenedioxy-β-nitrostyrene) is equally potent as Inhibitors of
platelet aggregation as 3,4-dimethoxy-β-nitrostyrene. MNS (20 μM)
concentration-dependently prevents ATP release from platelets stimulated
by thrombin or collagen. MNS (20 μM) inhibits thrombin-induced PAC-1
binding to human platelets.


The nitro group of MNS (3,4-Methylenedioxy-β-nitrostyrene) is essential for its antiplatelet effect.




Protocol(Only for Reference)


Kinase Assay: [1]

Tyrosine Kinase Assay

For immune complex kinase assay, platelets treated with MNS, PP1, or piceatannol are lysed and immunoprecipitated with either anti-Src or anti-Syk antibody. In some experiments, recombinant human Src, Syk, FAK, or JAK2 is used and treated with MNS or tyrosine kinase inhibitors for 5 min. The activity of the immunoprecipitated kinases or the recombinant enzymes is determined by using a tyrosine kinase assay kit. Samples are incubated with biotinylated poly[Glu:Tyr] (4:1), an exogenous substrate, in kinase assay buffer (20 mM HEPES, 5 mM MgCl2, 5 mM MnCl2, 0.1 mM sodium orthovanadate, and 1 mM dithiothreitol) for 37℃ for 30 to 45 min. After stopping the enzyme reaction with EDTA (final concentration of 20 mM), both the phosphorylated and dephosphorylated substrates are immobilized by binding to the streptavidin-coated plate. The fraction of phosphorylated substrate is determined using a phosphotyrosine monoclonal antibody conjugated to HRP and an ensuing chromogenic substrate reaction.

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

Species	Baboon	Dog	Monkey	Rabbit	Guinea pig	Rat	Hamster	Mouse
Weight (kg)	12	10	3	1.8	0.4	0.15	0.08	0.02
Body Surface Area (m2)	0.6	0.5	0.24	0.15	0.05	0.025	0.02	0.007
Km factor	20	20	12	12	8	6	5	3

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 Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×	mouse Km(3)	= 11.2 mg/kg
	rat Km(6)

Chemical Information

Molecular Weight (MW)	193.16
Formula	C9H7NO4
CAS No.	1485-00-3

Storage	3 years -20℃Powder
	6 months-80℃in solvent (DMSO, water, etc.)
Synonyms	N/A

Solubility (25°C) *	In vitro	DMSO	39 mg/mL (201.9 mM)
		Water	<1 mg/mL (
		Ethanol	<1 mg/mL (

Chemical Name	3,4-Methylenedioxy-β-nitrostyrene

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