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

Executive Summary: Dorsomorphin (Compound C) is a cell-permeable, ATP-competitive inhibitor of AMP-activated protein kinase (AMPK) with a Ki of 109 nM, exhibiting high selectivity over related kinases and robust inhibition of downstream targets such as ACC phosphorylation (80% reduction) and autophagic proteolysis. It also acts as a potent inhibitor of BMP/Smad signaling by blocking Smad1/5/8 phosphorylation, thereby modulating cellular differentiation and iron metabolism in preclinical models (Lei et al., 2024). Dorsomorphin is insoluble in water and ethanol but soluble in DMSO at ≥8.49 mg/mL with gentle warming and sonication, and is ideally used fresh due to limited solution stability. APExBIO supplies Dorsomorphin (SKU B3252) for research applications ranging from AMPK signaling studies in hepatocytes to modulation of BMP-induced gene expression and iron homeostasis. Recommended concentrations span 4–40 μM in cell culture and 10 mg/kg via intraperitoneal injection in animal models.

Biological Rationale

AMP-activated protein kinase (AMPK) is a central regulator of cellular energy homeostasis, modulating catabolic and anabolic pathways in response to metabolic stress (Lei et al., 2024). Dysregulated AMPK activity is implicated in metabolic disorders, inflammation, and cancer. In parallel, bone morphogenetic protein (BMP) signaling via Smad1/5/8 plays key roles in cell differentiation, embryogenesis, and iron metabolism. Inhibition of these pathways is crucial for probing mechanisms underlying autophagy, stem cell fate, and metabolic disease. Dorsomorphin's dual inhibitory profile makes it indispensable for dissecting AMPK and BMP/Smad pathway interactions in both in vitro and in vivo models (APExBIO).

Mechanism of Action of Dorsomorphin (Compound C)

Dorsomorphin (Compound C) acts as a reversible, ATP-competitive inhibitor of AMPK, binding to the kinase domain and blocking ATP access. Its Ki for AMPK is 109 nM, with pronounced selectivity over kinases such as protein kinase A, protein kinase C, and Janus kinase 3 (APExBIO). The compound reduces ACC phosphorylation by up to 80%, thereby suppressing fatty acid biosynthesis and autophagic proteolysis. Dorsomorphin also inhibits BMP signaling by preventing phosphorylation of Smad1/5/8, resulting in decreased hepcidin transcription and increased serum iron. In pluripotent stem cells, inhibition of BMP/Smad signaling by Dorsomorphin promotes self-renewal and neural induction. Its cell permeability enables both cell-based and animal studies, with rapid and reversible action.

Evidence & Benchmarks

• AMPK inhibition by Dorsomorphin (Compound C) (Ki = 109 nM) is selective over PKA, PKC, and JAK3 in kinase profiling assays (APExBIO).
• Dorsomorphin suppresses ACC phosphorylation by 80% in HeLa cells at 10 μM following AMPK activation (Lei et al., 2024).
• Inhibition of BMP4-induced SMAD1/5/8 phosphorylation (IC50 = 0.47 μM) observed in reporter assays (APExBIO).
• Animal models: 10 mg/kg intraperitoneal injection reduces hepatic hepcidin mRNA and elevates serum iron (Lei et al., 2024).
• Dorsomorphin induces dorsalization in zebrafish embryos by interfering with BMP signaling (APExBIO).
• AMPK inhibition via Dorsomorphin modulates M1 macrophage polarization and inflammatory factor production in LPS-treated murine macrophages (Lei et al., 2024).

For additional guidance on experimental applications and troubleshooting, see the Dorsomorphin (Compound C): Applied Strategies article, which details workflow optimization and extends the mechanistic depth presented here.

Applications, Limits & Misconceptions

Dorsomorphin (Compound C) is employed in research targeting:

• Inhibition of AMPK activity in hepatocytes and HeLa cells for metabolic and autophagy studies.
• Suppression of BMP4-induced SMAD phosphorylation in differentiation and iron metabolism models.
• Induction of neural differentiation and maintenance of embryonic stem cell pluripotency.
• Reduction of hepatic hepcidin expression and modulation of iron homeostasis in animal models.
• Interrogation of autophagic proteolysis and cell fate decisions.

This review clarifies and updates key data points relative to the Dorsomorphin: Precision AMPK Inhibitor article by explicitly benchmarking inhibition potencies and experimental constraints in diverse cell types and animal models.

Common Pitfalls or Misconceptions

• Not a pan-kinase inhibitor: While highly selective for AMPK and BMP/Smad, Dorsomorphin does not broadly inhibit unrelated kinases such as ERK, Akt, or MAPK at standard concentrations (APExBIO).
• Not suitable for long-term solution storage: Dorsomorphin rapidly loses activity in solution; freshly prepared DMSO stocks are recommended (APExBIO).
• Insoluble in aqueous buffers: The compound cannot be dissolved in water or ethanol; DMSO is required for stock solutions (≥8.49 mg/mL, with warming/sonication).
• Limited in vivo selectivity: Off-target effects may occur at high doses or in non-mammalian systems; dose titration is essential.
• Not a clinical drug: Dorsomorphin is for research use only and not approved for human or veterinary therapy.

For advanced cell model applications and protocol reproducibility, the Reliable AMPK and BMP Pathway Inhibition article provides specific troubleshooting scenarios, extending the practical scope of this dossier.

Workflow Integration & Parameters

• Reconstitution: Dissolve in DMSO at ≥8.49 mg/mL with gentle warming and ultrasonic treatment.
• Recommended concentrations: 4–40 μM in cell-based assays; 10 mg/kg for intraperitoneal injection in animal models.
• Storage: Solid at -20°C; avoid long-term storage of solutions.
• Controls: Include DMSO vehicle controls and, where possible, alternative AMPK/BMP inhibitors for specificity validation.
• Readouts: Western blot for ACC and SMAD phosphorylation, qPCR for hepcidin, ELISA for cytokines, and phenotypic readouts in zebrafish or stem cell models.

For comprehensive, scenario-based experimental design, refer to the Advanced AMPK and BMP Pathway Guide, which builds upon the present summary with workflow troubleshooting and protocol extensions.

Conclusion & Outlook

Dorsomorphin (Compound C), provided by APExBIO, is a validated, dual-pathway inhibitor enabling precise manipulation of AMPK and BMP/Smad signaling in metabolic, autophagy, and differentiation research. Its robust selectivity, well-characterized potency, and straightforward integration into cell and animal workflows make it an essential probe for dissecting molecular mechanisms of energy metabolism and cellular fate. Continued benchmarking and optimization will further expand its utility in disease modeling and drug discovery.