Dorsomorphin (Compound C): AMPK Inhibition and Mitochondrial Quality Control in Muscle and Stem Cell Research

Introduction

In cellular physiology and translational research, the intricate crosstalk between metabolic regulation, autophagy, and differentiation is a focal point for therapeutic innovation. Dorsomorphin (Compound C)—a highly selective, cell-permeable, ATP-competitive AMPK inhibitor (SKU: B3252)—has emerged as a foundational tool for dissecting these intertwined pathways. Its dual role as an AMPK and BMP signaling inhibitor makes it invaluable for studying autophagy regulation, neural stem cell differentiation, and iron metabolism modulation. While prior publications have spotlighted Dorsomorphin's dual-pathway inhibition and translational potential, this article delves deeper into its role as a modulator of mitochondrial quality control, especially in the context of muscle atrophy and stem cell fate, and strategically differentiates itself by integrating new mechanistic insights from recent literature.

Mechanism of Action of Dorsomorphin (Compound C)

AMPK Inhibition: Molecular Selectivity and Downstream Effects

Dorsomorphin (Compound C) is characterized by its high affinity for AMP-activated protein kinase (AMPK), acting as a reversible, ATP-competitive inhibitor with a Ki of 109 nM. Its selectivity for AMPK over related kinases—including protein kinase A, protein kinase C, and Janus kinase 3—enables precise modulation of the AMPK signaling pathway without off-target interference. By inhibiting AMPK activity, Dorsomorphin robustly suppresses phosphorylation of downstream substrates such as acetyl-CoA carboxylase (ACC), resulting in up to 80% reduction in ACC phosphorylation. This disruption impairs fatty acid oxidation, directly impacting cellular energy homeostasis and autophagic flux.

BMP/Smad Pathway Inhibition: Implications for Differentiation

Beyond metabolic signaling, Dorsomorphin is a potent BMP signaling inhibitor, notably blocking phosphorylation of Smad 1/5/8. This suppression of BMP4-induced SMAD phosphorylation (IC₅₀ ≈ 0.47 μM) has profound implications for developmental biology, including inhibition of heterotopic ossification, reduction of hepatic hepcidin transcription, and enhancement of neural induction in human embryonic stem cells. These features position Dorsomorphin as a dual-pathway modulator for applications ranging from disease modeling to regenerative medicine.

AMPK, Autophagy Regulation, and Mitochondrial Quality Control

The AMPK–Mitophagy Axis in Skeletal Muscle

The maintenance of mitochondrial integrity is vital for skeletal muscle function and adaptation. AMPK orchestrates energy sensing and is a pivotal regulator of autophagy, particularly mitophagy—the selective autophagic degradation of dysfunctional mitochondria. A recent study (Ren et al., 2025) elucidated the central role of AMPK/PINK1/Parkin-mediated mitophagy in mitigating skeletal muscle atrophy under high-fat diet-induced stress. In that work, the administration of Lycium barbarum polysaccharide (LBP) activated AMPK and promoted mitophagy, resulting in improved mitochondrial morphology, ATP production, and decreased oxidative stress in muscle tissue. Crucially, these protective effects were abrogated by AMPK inhibition—including the use of Dorsomorphin (Compound C)—and by Parkin knockdown, highlighting the necessity of AMPK activity for mitophagic quality control and muscle maintenance.

Experimental Evidence: Dorsomorphin as a Probe of Autophagy and Muscle Atrophy

Dorsomorphin’s unique ability to selectively inhibit AMPK has made it indispensable for dissecting the molecular underpinnings of autophagy regulation in both in vitro and in vivo models. In studies with hepatocytes and HeLa cells, Dorsomorphin impedes AMPK-driven phosphorylation events, resulting in the suppression of autophagic proteolysis. In animal models, its administration (typically at 10 mg/kg intraperitoneally) not only reduces hepatic hepcidin mRNA, impacting iron metabolism, but also serves as a critical negative control for delineating the necessity of AMPK activation in muscle health, as demonstrated by its capacity to block LBP-induced mitophagic rescue in muscle atrophy models (Ren et al., 2025).

Comparative Analysis: Dorsomorphin in Context of Alternative Approaches

While several articles have provided comprehensive overviews of Dorsomorphin’s dual-pathway inhibition and translational research value, such as the detailed mechanistic analysis found in "Strategic Dual-Pathway Inhibition: Advancing Translational Research", our focus diverges by exploring mitochondrial quality control as a central axis for interpreting Dorsomorphin's biological impact. Where previous works have emphasized the breadth of Dorsomorphin’s applications across metabolic, neural, and disease models, this article prioritizes the mechanistic interplay between AMPK inhibition, mitophagy, and skeletal muscle integrity—an emergent research frontier informed by the latest literature.

Alternative AMPK inhibitors and genetic knockdown strategies lack the rapid, reversible, and highly selective profile of Dorsomorphin, making it the preferred choice for time-sensitive or dose-responsive studies. Furthermore, Dorsomorphin’s dual inhibition profile allows simultaneous dissection of autophagy and differentiation pathways, making it uniquely suited for complex experimental designs where metabolic and developmental cues intersect.

Advanced Applications in Muscle, Neural, and Iron Metabolism Research

Dissecting Muscle Atrophy and Sarcopenic Obesity

The intersection of autophagy regulation and muscle health is exemplified in sarcopenic obesity, where obesity and muscle wasting co-occur. The ability of Dorsomorphin to block AMPK activity and suppress mitophagy serves as both a mechanistic probe and a potential therapeutic consideration. In the context of LBP-mediated rescue of muscle atrophy, Dorsomorphin administration enabled researchers to demonstrate that AMPK activation is a prerequisite for mitophagy-dependent muscle preservation (Ren et al., 2025). This highlights a translational application of Dorsomorphin: defining molecular checkpoints in metabolic disease and muscle degeneration.

Neural Stem Cell Differentiation and Regenerative Medicine

Dorsomorphin has proven instrumental in delineating the role of BMP/Smad signaling in stem cell biology. By inhibiting BMP-induced SMAD phosphorylation, it promotes the self-renewal and neural induction of human embryonic stem cells, enabling researchers to steer lineage commitment with high precision. This function underpins its use in neural stem cell differentiation protocols and regenerative medicine, providing a controllable axis for neurogenesis studies.

Iron Metabolism Modulation

Through its suppression of hepatic hepcidin gene transcription, Dorsomorphin modulates systemic iron homeostasis—an effect of clinical interest in anemia of chronic disease and iron overload disorders. This property is mechanistically distinct from its autophagy and differentiation roles, broadening its translational reach.

Experimental Considerations: Solubility, Dosing, and Handling

Dorsomorphin is insoluble in water and ethanol but dissolves readily in DMSO (≥8.49 mg/mL) with gentle warming and ultrasonic treatment. It is supplied as a solid and should be stored at -20°C. Solutions should be freshly prepared and used promptly, as long-term storage is not recommended. Optimal concentrations for cell culture studies range from 4 to 40 μM, while animal studies typically utilize 10 mg/kg via intraperitoneal injection. These parameters ensure consistent inhibition of AMPK activity and BMP/Smad signaling across experimental systems.

Integration with Existing Literature: Building a New Perspective

Whereas earlier articles such as "Dorsomorphin (Compound C): Precision AMPK and BMP Inhibition" provide an evidence-based overview of Dorsomorphin’s mechanisms and practical limitations, and "Strategic Leveraging of Dual AMPK and BMP Pathway Inhibition" frame it within the context of advanced disease models, this article distinguishes itself by: 1) focusing on mitochondrial quality control and mitophagy as central themes; 2) integrating recent experimental findings—especially the functional antagonism between Dorsomorphin and LBP in muscle atrophy; and 3) providing actionable guidance for leveraging Dorsomorphin to interrogate the AMPK–mitophagy axis in both muscle and neural contexts. This approach fills a critical gap in the content landscape, offering a mechanistically focused, application-driven synthesis for researchers seeking to unravel the complexities of metabolic and developmental regulation using Dorsomorphin.

Conclusion and Future Outlook

Dorsomorphin (Compound C) stands at the nexus of metabolic, autophagic, and developmental biology. Its unique profile as a selective ATP-competitive AMPK inhibitor and BMP/Smad pathway antagonist makes it an indispensable reagent for probing the molecular mechanisms underlying muscle atrophy, neural stem cell differentiation, and iron metabolism modulation. As demonstrated by recent research (Ren et al., 2025), Dorsomorphin’s ability to block AMPK-driven mitophagy provides a mechanistic gateway to understanding—and potentially manipulating—mitochondrial quality control in muscle and beyond.

Future directions include the integration of Dorsomorphin into combinatorial screening platforms, advanced disease models, and stem cell engineering protocols. As the interplay between metabolic signaling, autophagy regulation, and differentiation becomes increasingly central to therapeutic development, Dorsomorphin (Compound C) will remain a cornerstone tool for biomedical discovery.