A3963-10mg | A-769662 [844499-71-4] Clinisciences

Background

A-769662 is a potent activator of AMPK with EC50 value of 0.8 μM in vitro[1].

AMPK(AMP-activated protein kinase) is a serine/threonine protein kinase which is formed by three proteins: α,β and γ subunits. They play important roles in both the activity and stabilities of AMPK. AMPK functions as sensors of cellular energy by detecting and reacting to the change of AMP: ATP ratio. AMPK plays an important role in regulating intracellular energy metabolism. AMPK inhibits ATP-consuming pathways containing cholesterol synthesis, fatty acid synthesisand gluconeogenesis. AMPK stimulates fatty acid oxidation and glycolysis which are ATP-generating processes. AMPK inhibits gluconeogenesis by suppressing the expression of glucose-6-phosphatase and PEPCK wich are two key gluconeogenic enzymes.[1]

A-769662 belongs to the thienopyridone family. It activates the activity of purified AMPK from diffrent tissues with a dose-dependent manner. A-769662 activated the activity of AMPK extracted from human embryonic kidney cells (HEKs), rat muscle, or rat heart with EC50 values of 1.1 mM, 1.9 mM, or 2.2mM, respectively. A-769662 inhibited the synthesis of fatty acid with IC50 value of 3.2mM in primary rat hepatocytes.[1] A769662 also has inhibition effect on the 26S proteasome with an AMPK-independent mechanism. A769662 does not inhibit the proteolytic activities of the 20S core subunit which is a novel mechanism. A769662 can cause the arrest of cell cycle by inhibiting the 26S proteasome[2]. A-769662 activates AMPK in an allosterically manner. It also inhibits Thr-172 dephosphorylation of AMPK.[3]

In mice treated with dose of 30mg/kg, A-769662 decreased the expression of FAS, G6Pase, and PEPCK in liver of. It also lowered plasma glucose by 40% and reduced body weight gain.[1]

References:
[1]. Cool B, Zinker B, Chiou W, Kifle L, Cao N, Perham M, Dickinson R, Adler A, Gagne G, Iyengar R et al: Identification and characterization of a small molecule AMPK activator that treats key components of type 2 diabetes and the metabolic syndrome. Cell Metab 2006, 3(6):403-416.
[2]. Moreno D, Knecht E, Viollet B, Sanz P: A769662, a novel activator of AMP-activated protein kinase, inhibits non-proteolytic components of the 26S proteasome by an AMPK-independent mechanism. FEBS Lett 2008, 582(17):2650-2654.
[3]. Sanders MJ, Ali ZS, Hegarty BD, Heath R, Snowden MA, Carling D: Defining the mechanism of activation of AMP-activated protein kinase by the small molecule A-769662, a member of the thienopyridone family. J Biol Chem 2007, 282(45):32539-32548.

Product Citation

1. Ji-Man Park, Da-Hye Lee, et al. "Redefining the role of AMPK in autophagy and the energy stress response." Nat Commun. 2023 May 24;14(1):2994. PMID: 37225695
2. Pier, Marissa. "AMP-Activated Protein Kinase (AMPK) and Estrogen-Dependent Mechanisms Underlying Increased Susceptibility to Cardiovascular Disease During Menopause." The University of Arizona.
3. Yong Heui Jeon, Minzhen He, et al. "Adiponectin enhances the bioenergetics of cardiac myocytes via an AMPK-and succinate dehydrogenase-dependent mechanism." Cell Signal. 2021 Feb;78:109866. PMID: 33271223
4. Tereza Moore, Rolando E. Yanes, et al. "AMP-independent activator of AMPK for treatment of mitochondrial disorders." PLoS One. 2020 Oct 14;15(10):e0240517. PMID: 33052980
5. Matzinger M, Fischhuber K, et al. "AMPK leads to phosphorylation of the transcription factor Nrf2, tuning transactivation of selected target genes." Redox Biol. 2019 Nov 27;29:101393. PMID: 31805502
6. Cannon, Danielle Kathryn. "ASSESSING MENOPAUSAL FEMALE SUSCEPTIBILITY TO HEART DISEASE: A FOCUS ON AMPK’S ABILITY TO MITIGATE CARDIAC REMODELING THROUGH ESTROGEN-DEPENDENT GENE PROGRAMS." The University of Arizona. 2019.
7. Wen Z, Jin K, et al. "N-myristoyltransferase deficiency impairs activation of kinase AMPK and promotes synovial tissue inflammation." Nat Immunol. 2019 Mar;20(3):313-325. PMID: 30718913
8. Hubbard JA, Xiao B, et al. "Production and Crystallization of Full-Length Human AMP-Activated Protein Kinase (α1β1γ1)." Methods Mol Biol. 2018;1732:1-14. PMID: 29480465
9. Oladimeji PO, Lin W, et al. "Glucose-dependent regulation of pregnane X receptor is modulated by AMP-activated protein kinase." Sci Rep. 2017 Apr 24;7:46751. PMID: 28436464

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Chemical Properties

Physical Appearance	A solid
Storage	Store at -20°C
M.Wt	360.39
Cas No.	844499-71-4
Formula	C₂₀H₁₂N₂O₃S
Synonyms	A-769662; A769662
Solubility	insoluble in EtOH; insoluble in H2O; ≥18.02 mg/mL in DMSO
Chemical Name	4-hydroxy-3-[4-(2-hydroxyphenyl)phenyl]-6-oxo-7H-thieno[2,3-b]pyridine-5-carbonitrile
SDF	Download SDF
Canonical SMILES	C1=CC=C(C(=C1)C2=CC=C(C=C2)C3=CSC4=C3C(=C(C(=O)N4)C#N)O)O
Shipping Condition	Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips	We do not recommend long-term storage for the solution, please use it up soon.

Protocol

Cell experiment [1]:
Cell lines	Primary rat hepatocytes.
Preparation method	Soluble in DMSO >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.
Reaction Conditions	4 h.
Applications	A-769662 dose-dependently increases ACC phosphorylation, which is phosphorylated by AMPK. A-769662 inhibits fatty acid synthesis with IC50 value of 3.2 μM. Treatment of rat hepatocytes with A-769662 at concentrations up to 100 μM shows no measurable cytotoxicity.
Animal experiment [1]:
Animal models	Sprague Dawley (SD) rats.
Dosage form	30 mg/kg; gavaged.
Applications	A-769662 significantly reduces the respiratory exchange ratio (RER) throughout the first 3 h, which is followed by a small but significant increase in RER over the subsequent 3 h. A-769662 reduces malonyl CoA levels in rat livers by 33%.
Other notes	Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
References: [1]. Cool B, Zinker B, Chiou W, et al. Identification and characterization of a small molecule AMPK activator that treats key components of type 2 diabetes and the metabolic syndrome. Cell Metab, 2006, 3(6): 403-416.