Dorsomorphin (Compound C): ATP-Competitive AMPK and BMP Signaling Inhibitor

Executive Summary: Dorsomorphin (Compound C) is a cell-permeable, ATP-competitive inhibitor with a Ki of 109 nM for AMPK, showing high selectivity over related kinases (APExBIO, product page). It inhibits downstream phosphorylation of acetyl-CoA carboxylase (ACC) by 80% and suppresses autophagic proteolysis (Yu et al., 2008, DOI). Dorsomorphin blocks BMP signaling by inhibiting Smad 1/5/8 phosphorylation, reducing hepcidin transcription and increasing serum iron (Yu et al., 2008, DOI). It is insoluble in water/ethanol, dissolves in DMSO at ≥8.49 mg/mL, and is supplied as a solid for -20°C storage (APExBIO, B3252 kit). Animal studies confirm efficacy in modulating iron metabolism and hepcidin mRNA levels (Yu et al., 2008, DOI).

Biological Rationale

AMP-activated protein kinase (AMPK) is a master regulator of cellular energy homeostasis. It responds to metabolic stress by promoting catabolic pathways and suppressing anabolic processes (Hardie, 2011, DOI). Dysregulation of AMPK is implicated in metabolic diseases, cancer, and neurodegeneration. The BMP/Smad pathway controls cell fate, differentiation, and iron metabolism (Yu et al., 2008, DOI). Dorsomorphin enables targeted inhibition of both AMPK and BMP/Smad signaling, supporting research on energy metabolism, autophagy regulation, and developmental biology (see prior analysis; this article extends by providing updated mechanistic benchmarks and evidence).

Mechanism of Action of Dorsomorphin (Compound C)

Dorsomorphin is a reversible, ATP-competitive inhibitor of AMPK with a Ki of 109 nM, showing high selectivity over protein kinase A, kinase C, and Janus kinase 3 (Yu et al., 2008, DOI). It blocks the phosphorylation of ACC, a direct AMPK substrate, by 80% in vitro at micromolar concentrations. Dorsomorphin also suppresses autophagic proteolysis, acting downstream of AMPK inhibition (Zhou et al., 2001, DOI). Beyond AMPK, dorsomorphin inhibits BMP signaling by blocking SMAD 1/5/8 phosphorylation with an IC50 of 0.47 μM (Yu et al., 2008, DOI). This dual action allows precise modulation of metabolic and developmental pathways.

Evidence & Benchmarks

• Dorsomorphin inhibits AMPK activity in hepatocytes and HeLa cells with a Ki of 109 nM (Yu et al., 2008, DOI).
• Suppresses ACC phosphorylation by 80% at 10 μM in cultured cells (Yu et al., 2008, DOI).
• Inhibits BMP4-induced SMAD phosphorylation with an IC50 of 0.47 μM (Yu et al., 2008, DOI).
• Reduces hepatic hepcidin mRNA and increases serum iron in animal models (Yu et al., 2008, DOI).
• Promotes self-renewal and neural induction in human embryonic stem cells by inhibiting BMP pathways (Chambers et al., 2009, DOI).
• Induces dorsalization in zebrafish embryos, a canonical BMP pathway phenotype (Yu et al., 2008, DOI).
• Recommended concentrations: 4–40 μM in cell culture; 10 mg/kg i.p. in animal models (APExBIO, B3252 product).
• Dorsomorphin is insoluble in water/ethanol, soluble in DMSO at ≥8.49 mg/mL (APExBIO, product page).

For additional mechanistic discussion and translational use-cases, see this article, which we extend here with updated solubility and storage parameters relevant for workflow integration.

Applications, Limits & Misconceptions

Dorsomorphin (Compound C) is widely used to dissect the AMPK signaling pathway, study autophagy regulation, and modulate BMP/Smad signaling in developmental and metabolic research. Its dual inhibition of AMPK and BMP makes it a key reagent for exploring iron metabolism, neural stem cell differentiation, and cancer cell metabolism (detailed benchmarking; this article provides updated guidelines for dosing and experimental design).

Common Pitfalls or Misconceptions

• Dorsomorphin is not selective for AMPK at high concentrations (>40 μM) and may inhibit off-target kinases.
• It does not inhibit all BMP family members equally; potency varies across ligands.
• Insolubility in water/ethanol limits its use in some aqueous assays—DMSO is required for stock solutions.
• Long-term storage of solutions is not recommended; activity declines over time even at -20°C.
• Does not directly activate or inhibit Nrf2/Keap1 pathway, but can modulate stress response indirectly via metabolic changes (Patra et al., 2020, DOI).

Workflow Integration & Parameters

Dorsomorphin is supplied as a solid, best stored at -20°C in a desiccated environment. For experimental use, dissolve in DMSO at concentrations ≥8.49 mg/mL with gentle warming and ultrasonic treatment. Use at 4–40 μM for cell-based assays and 10 mg/kg via intraperitoneal injection for animal studies. Prepare working solutions fresh; avoid prolonged storage. APExBIO recommends prompt use after preparation (product page).

For precise AMPK or BMP/Smad pathway interrogation, time-dependent and concentration-dependent controls are essential. Use validated antibodies for ACC and SMAD phosphorylation endpoints. For autophagy studies, monitor LC3-II conversion and p62 degradation (advanced mechanistic insights; this article adds practical workflow parameters and storage tips).

Conclusion & Outlook

Dorsomorphin (Compound C) remains a gold-standard tool for dissecting AMPK and BMP/Smad signaling in metabolic, cancer, and developmental biology. Its dual specificity, characterized benchmarks, and robust performance in both cell and animal models make it indispensable for translational research. APExBIO's quality assurance ensures reproducibility across laboratories. Ongoing studies continue to expand its applications in neural differentiation, iron metabolism, and precision disease modeling.