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Chemical Structure| 1261289-04-6 Chemical Structure| 1261289-04-6

Structure of O-304
CAS No.: 1261289-04-6

Chemical Structure| 1261289-04-6

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O-304 is a first-in-class, orally available pan-AMPK activator, which increases AMPK activity by suppressing the dephosphorylation of pAMPK. O-304 exhibits a great potential as a drug to treat type 2 diabetes (T2D) and associated cardiovascular complications [1][2].

Synonyms: O-304X

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Product Details of O-304

CAS No. :1261289-04-6
Formula : C16H11Cl2N3O2S
M.W : 380.25
SMILES Code : ClC1=CC=C(C=C1)CN2C(N=C(NC(C3=CC=C(Cl)C=C3)=O)S2)=O
Synonyms :
O-304X
MDL No. :MFCD31657425
InChI Key :WEDWLYRQKUTOAX-UHFFFAOYSA-N
Pubchem ID :50923806

Safety of O-304

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H315-H319-H335
Precautionary Statements:P261-P280-P301+P312-P302+P352-P305+P351+P338

Related Pathways of O-304

epigenetics
PI3K-AKT

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
mouse primary islets 2.5 μM, 5 μM, 10 μM O304 increased p-T172 AMPK levels JCI Insight. 2018 Jun 21;3(12):e99114.
INS-1 insulinoma cells 2.5 μM, 5 μM, 10 μM O304 increased p-T172 AMPK levels JCI Insight. 2018 Jun 21;3(12):e99114.
hepatocytes 2.5 μM, 5 μM, 10 μM O304 dose-dependently suppressed lipid synthesis JCI Insight. 2018 Jun 21;3(12):e99114.
Human skeletal myotubes 2.5 μM, 5 μM, 10 μM O304 increased 2-DeoxyD-glucose (2-DG) uptake in a dose- and AMPK-dependent manner in the absence of insulin JCI Insight. 2018 Jun 21;3(12):e99114.
Wi-38 human lung fibroblast cells 2.5 μM, 5 μM, 10 μM O304 increased pAMPK, pACC, and ATP/protein ratio in a dose-dependent manner JCI Insight. 2018 Jun 21;3(12):e99114.
INS-1E cells 5 μM 4 days or 2 h To evaluate the effect of O304 on insulin secretion under hyperglycemic conditions, results showed that O304 prevented and partly reversed the negative effects of chronic hyperglycemia on glucose-stimulated insulin secretion (GSIS). Commun Biol. 2023 Aug 25;6(1):877.
C2C12 myotubes 0.625, 1.25, 2.5 μM 4 h To assess the uncoupling potential of O304, results showed that O304 dose-dependently increased oxygen consumption rate (OCR), indicating its function as a mitochondrial uncoupler. Commun Biol. 2023 Aug 25;6(1):877.
mouse hippocampal neurons 50 nM 1 h Pre-treatment with O304 abrogated the suppression effect of NTRK1 knockdown on the activity of the AMPK/ULK1/FUNDC1 pathway and reversed the inhibitory effect of NTRK1 knockdown on mitophagy. Cell Death Discov. 2023 Oct 31;9(1):404.
Human aortic smooth muscle cells 50 nM 48 h O304 activated AMPK signaling and prevented VSMC phenotypic switching from the contractile to the synthetic phenotype. Front Pharmacol. 2024 Nov 29;15:1457817.
CBA x C57Bl/6 F1 hybrid mouse islets 5 μM 96 h O304 significantly reduced Aldh1a3 expression under high glucose conditions, alleviating β-cell stress Sci Rep. 2021 Dec 23;11(1):24410.

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
Mice Diet-induced obese (DIO) mice Oral gavage 100 mg/kg Once daily for 8 weeks O304 and O304+Metformin significantly reduced fasted glucose and plasma insulin levels and prevented insulin resistance JCI Insight. 2018 Jun 21;3(12):e99114.
Mice STZ-induced diabetic mice and db/db mice Formulated in diet 0.25, 0.5, 1.0 mg/g Continuous treatment To assess the ameliorative effects of O304 on hyperglycemia and its protective effects on β-cell function. Results showed that O304 significantly improved hyperglycemia by dually promoting muscle glucose effectiveness and preserving β-cell function. Commun Biol. 2023 Aug 25;6(1):877.
ApoE-deficient male mice Abdominal aortic aneurysm model Oral 200 mg/kg Once every 3 days for 28 days O304 significantly activated AMPK signaling, increased the proportion of contractile VSMCs, and reduced AAA formation and blood pressure. Front Pharmacol. 2024 Nov 29;15:1457817.
Mice Aged mouse model Oral 0.25 or 0.5 mg/g Continued for 12 months O304 prevented and reverted age-associated hyperinsulinemia and insulin resistance, and improved cardiac function and exercise capacity in aged mice. Commun Biol. 2021 Nov 18;4(1):1306
CBA mice High-fat diet-induced obesity and diabetes model Oral 0.8 mg/g Continuous for 9 weeks O304 treatment significantly improved glucose homeostasis, reduced fasting blood glucose and insulin levels, alleviated β-cell stress, and restored islet gene expression profiles Sci Rep. 2021 Dec 23;11(1):24410.

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

2.63mL

0.53mL

0.26mL

13.15mL

2.63mL

1.31mL

26.30mL

5.26mL

2.63mL

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