AMPK activator 4 is a potent AMPK activator and does not inhibit mitochondrial complex I. AMPK activator 4 selectively activates AMPK in muscle tissue. AMPK activator 4 dose-dependently improved glucose tolerance in normal mice, significantly reduced fasting blood glucose levels and improved insulin resistance in db/db diabetic mice. Has hypoglycemic effect.

Physicochemical Properties

Molecular Formula	C24H21CLN2O3
Molecular Weight	420.888145208359
Exact Mass	420.124
CAS #	2493239-46-4
PubChem CID	154573764
Appearance	White to off-white solid powder
LogP	5.2
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	7
Heavy Atom Count	30
Complexity	593
Defined Atom Stereocenter Count	0
SMILES	ClC1C=CC(=CC=1)OC(C(NC1C=CC(=CC=1)OCC1C=CC(C#N)=CC=1)=O)(C)C
InChi Key	OSSMPMZJYDZMFX-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H21ClN2O3/c1-24(2,30-22-11-7-19(25)8-12-22)23(28)27-20-9-13-21(14-10-20)29-16-18-5-3-17(15-26)4-6-18/h3-14H,16H2,1-2H3,(H,27,28)
Chemical Name	2-(4-chlorophenoxy)-N-[4-[(4-cyanophenyl)methoxy]phenyl]-2-methylpropanamide
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	AMPK activator 4 (compound B10) (0–20 μM; 12–24 hours) causes AMPK and its downstream protein ACC to become phosphorylated[1]. LKB1 is necessary for AMPK phosphorylation mediated by AMPK activator 4[1]. When HepG2 cells are treated with AMPK activator 4 at 10–20 μM for 72 hours, their viability is slightly decreased[1].
ln Vivo	AMPK activator 4 (100 mg/kg; administered intragastrically; daily for 9 weeks) exhibits anti-hyperglycemic properties[1].
Cell Assay	Western Blot Analysis[1] Cell Types: C2C12 myotubes, HepG2 and HuH-7 cells Tested Concentrations: 0-20 μM Incubation Duration: 12-24 hrs (hours) Experimental Results: Induced dose-dependently phosphorylation of ACC in C2C12 myotube cells, and the phosphorylation of ACC was highly consistent with the phosphorylation of AMPK. Increased the phosphorylation of AMPK and ACC in human hepatocarcinoma HepG2 and HuH-7 cells.
Animal Protocol	Animal/Disease Models: db/db mice (5-week old)[1] Doses: 100 mg/kg Route of Administration: intragastric (po)administration; daily for 9 weeks Experimental Results: Dramatically lowered the fasting blood glucose in db/db mice (2-6 weeks). Dramatically improved insulin resistance in db /db mice (for 9 weeks). The body weight and the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (two markers of liver injury) were not Dramatically affected.
References	[1]. Discovery of AdipoRon analogues as novel AMPK activators without inhibiting mitochondrial complex I. Eur J Med Chem. 2020;200:112466.

Solubility Data

Solubility (In Vitro)

DMSO : 250 mg/mL (593.98 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3759 mL	11.8796 mL	23.7592 mL
	5 mM	0.4752 mL	2.3759 mL	4.7518 mL
	10 mM	0.2376 mL	1.1880 mL	2.3759 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.