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

Executive Summary: Dorsomorphin (Compound C) is a reversible, ATP-competitive inhibitor of AMP-activated protein kinase (AMPK) with a Ki of 109 nM, showing high selectivity over related kinases (ApexBio). It inhibits AMPK-dependent phosphorylation of acetyl-CoA carboxylase (ACC) by 80% at recommended concentrations, suppresses BMP/Smad1/5/8 signaling, and modulates hepatic hepcidin expression to increase serum iron (Ren et al., 2025). Dorsomorphin is insoluble in water/ethanol but dissolves in DMSO at ≥8.49 mg/mL with mild warming. It is widely used in muscle metabolism, neural stem cell differentiation, and autophagy/mitophagy studies. Limitations include incomplete specificity at high doses and rapid solution degradation, requiring prompt use post-reconstitution (ApexBio).

Biological Rationale

AMPK is a central regulator of cellular energy homeostasis, activated by increased AMP/ATP ratios. It phosphorylates downstream targets, including ACC and components of autophagy machinery, to restore energy balance (Ren et al., 2025). Dysregulation of AMPK is linked to metabolic syndromes, cancer, and muscle atrophy. Bone morphogenetic protein (BMP) signaling also regulates cell differentiation, iron metabolism, and tissue patterning. Dorsomorphin (Compound C) was developed to selectively block AMPK and BMP/Smad pathways, enabling precise experimental dissection of their roles in metabolism, autophagy, and cell fate determination.

Mechanism of Action of Dorsomorphin (Compound C)

• AMPK Inhibition: Dorsomorphin binds the ATP-binding pocket of AMPK, competitively inhibiting its kinase activity with a Ki value of 109 nM (ApexBio).
• Downstream Effects: Inhibition suppresses phosphorylation of ACC (by up to 80%), reduces autophagic flux, and impairs PINK1/Parkin-mediated mitophagy in skeletal muscle models (Ren et al., 2025).
• BMP/Smad Signaling Inhibition: Dorsomorphin blocks BMP-induced phosphorylation of Smad1/5/8, with an IC₅₀ of 0.47 μM for BMP4-induced signaling in cell-based assays (ApexBio).
• Iron Metabolism Modulation: By suppressing hepatic hepcidin gene transcription, Dorsomorphin increases serum iron in animal models (Ren et al., 2025).
• Stem Cell Effects: Inhibition of BMP pathways by Dorsomorphin promotes self-renewal and neural induction in human embryonic stem cells (ApexBio).

Evidence & Benchmarks

• Dorsomorphin inhibits AMPK activity in hepatocytes and HeLa cells at 4–40 μM, with 80% reduction in ACC phosphorylation (ApexBio, product page).
• In mouse models, Dorsomorphin at 10 mg/kg intraperitoneally reduces hepatic hepcidin mRNA and increases serum iron (Ren et al., 2025, DOI).
• Dorsomorphin inhibits BMP4-induced SMAD1/5/8 phosphorylation with an IC₅₀ of 0.47 μM in cell culture (ApexBio, product page).
• Application of Dorsomorphin in zebrafish embryos induces dorsalization, demonstrating robust BMP inhibition (ApexBio, product page).
• In muscle atrophy models, AMPK inhibition with Dorsomorphin blocks LBP-induced mitophagy, confirming its action on the AMPK/PINK1/Parkin axis (Ren et al., 2025, DOI).

This article extends the mechanistic detail presented in Dorsomorphin (Compound C): Precision AMPK and BMP Inhibit... by integrating new evidence on iron metabolism modulation and storage/solubility parameters.

For a deeper exploration of Dorsomorphin's impact on mitophagy and muscle metabolism, see Dorsomorphin (Compound C): Precision AMPK Inhibition for ..., which this article augments by providing updated dosing benchmarks and workflow integration.

Applications, Limits & Misconceptions

Key Research Applications

• Inhibition of AMPK signaling in hepatocytes, skeletal muscle, and cancer models.
• Suppression of autophagy and mitophagy for metabolic disease modeling.
• Dissection of BMP/Smad signaling in stem cell differentiation and developmental biology.
• Modulation of iron metabolism via hepatic hepcidin regulation.
• Neural induction in human pluripotent stem cells through BMP pathway inhibition.

Common Pitfalls or Misconceptions

• Dorsomorphin is not a pan-kinase inhibitor: it shows high selectivity but may affect off-target kinases at elevated concentrations (≥40 μM).
• It is not suitable for long-term solution storage; activity degrades rapidly in DMSO at room temperature.
• Dorsomorphin is insoluble in water or ethanol, requiring DMSO and gentle warming for use.
• The compound does not universally block all forms of autophagy; effects are context-dependent and most robust in AMPK-dominant pathways.
• It does not substitute for genetic knockdown of AMPK or BMP signaling components, especially in chronic or developmental studies.

Workflow Integration & Parameters

• Reconstitution: Dissolve Dorsomorphin (B3252) in DMSO at ≥8.49 mg/mL with mild warming and ultrasonic treatment (ApexBio).
• Storage: Store solid at –20°C; use solutions promptly after preparation. Do not freeze/thaw solutions repeatedly.
• Recommended Usage: 4–40 μM in cell cultures; 10 mg/kg via intraperitoneal injection for animal models.
• Controls: Include vehicle (DMSO) and/or positive pathway inhibitors in all experimental designs.
• Readouts: Monitor phosphorylation markers (ACC, SMAD1/5/8), autophagic flux (LC3-II/I ratio), and iron metabolism markers (hepcidin mRNA, serum iron).

For strategic dual-pathway inhibition protocols, see Strategic Dual-Pathway Modulation: Dorsomorphin (Compound...); this article provides unique storage and dosing caveats not previously detailed.

Conclusion & Outlook

Dorsomorphin (Compound C) remains a gold-standard, reversible ATP-competitive inhibitor for the AMPK and BMP/Smad signaling pathways. Its use has elucidated the role of AMPK in autophagy, energy metabolism, and muscle atrophy models, and clarified BMP signaling in stem cell fate and iron metabolism. While highly selective at recommended doses, practitioners should be aware of solubility and stability constraints, and verify specificity in each system. Continued deployment of Dorsomorphin will advance metabolic disease modeling, stem cell biology, and translational research, provided strict adherence to validated protocols and controls. For product details and ordering, refer to the Dorsomorphin (Compound C) B3252 kit.