RSVA405 is a potent and orally bioactive AMPK activator with EC50 of 1 μM. RSVA405 inhibits mTOR by promoting CaMKKβ-dependent AMPK activation, and promotes autophagy to increase Aβ degradation. RSVA405 has anti-inflammatory effects by inhibiting STAT3 function. RSVA405 can also be used in obesity research.

Physicochemical Properties

Molecular Formula	C17H20N4O2
Molecular Weight	312.37
Exact Mass	312.159
CAS #	140405-36-3
PubChem CID	135400298
Appearance	Light yellow to yellow solid powder
LogP	2.788
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	23
Complexity	392
Defined Atom Stereocenter Count	0
SMILES	CCN(CC)C1=CC(=C(C=C1)/C=N/NC(=O)C2=CC=NC=C2)O
InChi Key	GWQPCBPAOAFXSJ-XDHOZWIPSA-N
InChi Code	InChI=1S/C17H20N4O2/c1-3-21(4-2)15-6-5-14(16(22)11-15)12-19-20-17(23)13-7-9-18-10-8-13/h5-12,22H,3-4H2,1-2H3,(H,20,23)/b19-12+
Chemical Name	N-[(E)-[4-(diethylamino)-2-hydroxyphenyl]methylideneamino]pyridine-4-carboxamide
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	AMPK 1 μM (EC50, in cell-based assays)
ln Vitro	RSVA405 (0.2-2 μM; 24 h) suppresses the development of adipocytes[2]. In 3T3-L1 cells, the production of fatty acid synthase (FAS), PPAR-γ, and fatty acid binding protein 4 (aP2) is strongly inhibited by RSVA405 (0.2-2 μM; 24 h)[2]. In activated RAW 264.7 macrophages, RSVA405 (1-3 μM; 16 h) suppresses LPS-induced STAT3 activity, intracellular signaling, and cytokine response[3]. With an EC50 of approximately 1 μM in APP-HEK293 cells, RSVA405 (1-3 μM; 24 h) suppresses mTOR, induces autophagy, and promotes the lysosomal breakdown of Aβ[4].
ln Vivo	In rats undergoing ischemia-reperfusion (I/R), RSVA405 (3 mg/kg; ip) reduces kidney damage and preserves renal function[1]. Mice on a high-fat diet have significantly less body weight gain when given RSVA405 (20–100 mg/kg/d; po for 11 weeks)[2].
Cell Assay	Cell Viability Assay[2] Cell Types: 3T3-L1 preadipocytes Tested Concentrations: 0.2, 0.5, 1, 2 μM Incubation Duration: 24 h Experimental Results: Increased the phosphorylation of AMPK and its substrate acetyl- CoA carboxylase (ACC). Inhibited the accumulation of lipid droplets in a dose-dependent manner, with an IC50 of 0.5 μM.
Animal Protocol	Animal/Disease Models: Male SD (Sprague-Dawley) rats (300-350 g) are induced I/ R injury[1] Doses: 3 mg/kg Route of Administration: Ip one hour before inducing I/R injury Experimental Results: diminished the levels of creatinine and blood urea nitrogen (BUN), by 35.8% and 44.3% in serum, respectively. diminished the levels of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) by 33.0% and 59.8% in serum, respectively.
References	[1]. Novel resveratrol analogues attenuate renal ischemic injury in rats. J Surg Res. 2015 Feb;193(2):807-15. [2]. Small-molecule activators of AMP-activated protein kinase (AMPK), RSVA314 and RSVA405, inhibit adipogenesis. Mol Med. Sep-Oct 2011;17(9-10):1022-30. [3]. Identification of potent small-molecule inhibitors of STAT3 with anti-inflammatory properties in RAW 264.7 macrophages. FEBS J. 2012 Oct;279(20):3791-9. [4]. Novel synthetic small-molecule activators of AMPK as enhancers of autophagy and amyloid-β peptide degradation. FASEB J. 2011 Jan;25(1):219-31.

Solubility Data

Solubility (In Vitro)

DMSO : 125 mg/mL (400.17 mM)

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	3.2013 mL	16.0067 mL	32.0133 mL
	5 mM	0.6403 mL	3.2013 mL	6.4027 mL
	10 mM	0.3201 mL	1.6007 mL	3.2013 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.