N-Acetyldopamine dimer-2 (compound 2) is an N-acetyldopamine dimer with antioxidant and anti-inflammatory activity that can be extracted from the yellow powder of cicada bark. N-Acetyldopamine dimer-2 inhibits oxidized low-density lipoprotein (LDL) oxidation, ROS generation, NO production, and NF-κB activity.

Physicochemical Properties

Molecular Formula	C20H20N2O6
Molecular Weight	384.382605552673
Exact Mass	384.132
CAS #	916888-49-8
PubChem CID	16072146
Appearance	Typically exists as solid at room temperature
LogP	1.5
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	597
Defined Atom Stereocenter Count	2
SMILES	CC(=O)N/C=C/C1=CC2=C(C=C1)O[C@@H]([C@H](O2)C3=CC(=C(C=C3)O)O)NC(=O)C
InChi Key	RCEWLGWTZHROAQ-CLJKYYQGSA-N
InChi Code	InChI=1S/C20H20N2O6/c1-11(23)21-8-7-13-3-6-17-18(9-13)27-19(20(28-17)22-12(2)24)14-4-5-15(25)16(26)10-14/h3-10,19-20,25-26H,1-2H3,(H,21,23)(H,22,24)/b8-7+/t19-,20+/m1/s1
Chemical Name	N-[(E)-2-[(2S,3R)-2-acetamido-3-(3,4-dihydroxyphenyl)-2,3-dihydro-1,4-benzodioxin-6-yl]ethenyl]acetamide
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

Targets	IC50: 1.5 μM (copper-mediated LDL oxidation), 2.7 μM (AAPH-mediated LDL oxidation), 1.6 μM (SIN-1-mediated LDL oxidation)[1]
ln Vitro	With IC50 values of 1.5, 2.7, and 1.6 μM, respectively, N-acetyldopamine dimer-2 inhibits copper-mediated, 2,2'azobis(2-amidinopropane) hydrochloride (AAPH)-mediated, and 3-morpholinosydnonimine (SIN)-1-mediated LDL oxidation[1]. N-Acetyldopamine dimer-2 exhibits a potent ability to scavenge radicals at 100 μM for 40 minutes[1]. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells exhibit a dose-dependent reduction in ROS when exposed to N-acetyldopamine dimer-2 (0-400 μM; 2 h)[1]. In LPS-stimulated RAW264, N-acetyldopamine dimer-2 (0-400 μM; 2 h) dose-dependently and slightly reduces NO production and iNOS protein expression. 7 cells [1]. N-acetyldopamine dimer-2 (0-400 μM; 2 h) suppresses IL-6, TNF-α, and COX-2 mRNA levels, and it also inhibits IL-6 secretion and LPS-induced NF-κB activation in RAW264.7 cells in a dose-dependent manner[1].
Cell Assay	Western Blot Analysis[1] Cell Types: LPS-stimulated RAW264.7 cell line Tested Concentrations: 200 and 400 μM Incubation Duration: 2 hrs (hours) Experimental Results: Slightly diminished iNOS protein expression in LPS-stimulated RAW264.7 cells. Western Blot Analysis[1] Cell Types: LPS-stimulated RAW264.7 cell line Tested Concentrations: 200 and 400 μM Incubation Duration: 2 hrs (hours) Experimental Results: Dose-dependently diminished the mRNA levels of IL-6, TNF-α and COX-2.
References	[1]. Antioxidant and anti-inflammatory activities of N-acetyldopamine dimers from Periostracum Cicadae. Bioorg Med Chem. 2006 Dec 1;14(23):7826-34.

Solubility Data

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.6016 mL	13.0080 mL	26.0159 mL
	5 mM	0.5203 mL	2.6016 mL	5.2032 mL
	10 mM	0.2602 mL	1.3008 mL	2.6016 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.