Y-27632 dihydrochloride
[CAS# 146986-50-7]

Identification

• Classification: Biochemical >> Inhibitor >> Cell cycle >> ROCK inhibitor
• Name: Y-27632 dihydrochloride
• Synonyms: 4-[(1R)-1-Aminoethyl]-N-pyridin-4-ylcyclohexane-1-carboxamide dihydrochloride
• Molecular Formula: C₁₄H₂₃Cl₂N₃O
• Molecular Weight: 320.26
• CAS Registry Number: 146986-50-7
• EC Number: 620-446-5
• SMILES: C[C@H](C1CCC(CC1)C(=O)NC2=CC=NC=C2)N

Properties

• Solubility: DMSO 3 mg/mL, Water <1 mg/mL, Ethanol 4 mg/mL (Expl.)

Safety Data

• Hazard Symbols: GHS07 Warning
• Hazard Statements: H302-H312-H332
• Precautionary Statements: P280

Hazard Classification

Hazard	Class	Category Code	Hazard Statement
Acute toxicity	Acute Tox.	4	H302
Chronic hazardous to the aquatic environment	Aquatic Chronic	1	H410

Discovory and Applicatios

Y-27632 dihydrochloride is a small-molecule compound that functions as a selective inhibitor of Rho-associated protein kinases (ROCKs), specifically ROCK1 and ROCK2. These kinases are downstream effectors of the small GTPase RhoA and play crucial roles in various cellular processes including actin cytoskeleton organization, cell migration, proliferation, apoptosis, and smooth muscle contraction. The development and characterization of Y-27632 provided researchers with a valuable pharmacological tool to study the Rho/ROCK signaling pathway, which has been implicated in numerous physiological and pathological conditions.

The compound was first identified and developed in the late 1990s through efforts to discover selective inhibitors of ROCK. Biochemical screening led to the synthesis of Y-27632, which demonstrated high affinity and selectivity for ROCK over other kinases. Its mechanism of action involves competitive binding to the ATP-binding site of the kinase domain, resulting in inhibition of ROCK enzymatic activity. This action was shown to effectively block ROCK-mediated phosphorylation of target substrates such as myosin light chain and LIM kinase, leading to disruption of actin stress fibers and inhibition of cellular contraction.

Y-27632 dihydrochloride has been extensively used in basic research to delineate the role of ROCK in cellular and developmental biology. In vitro studies revealed that treatment with Y-27632 promotes the survival and proliferation of dissociated human embryonic stem cells and induced pluripotent stem cells, which are otherwise prone to apoptosis upon single-cell dissociation. This application has significantly improved protocols for stem cell culture and has facilitated the expansion and manipulation of pluripotent cells for various downstream applications.

In addition to stem cell biology, Y-27632 has been used to investigate the role of ROCK in cancer, cardiovascular disease, and neurodegenerative disorders. In cancer research, it has helped elucidate the contribution of ROCK signaling to tumor cell migration, invasion, and metastasis. ROCK inhibitors like Y-27632 have demonstrated the ability to impair the motility and invasive potential of cancer cells in vitro, suggesting potential therapeutic implications in limiting cancer progression.

In cardiovascular studies, Y-27632 has been employed to explore its vasodilatory effects due to ROCK's role in regulating vascular tone. By inhibiting ROCK, Y-27632 can reduce smooth muscle contraction and lower blood pressure in experimental models, highlighting the involvement of Rho/ROCK signaling in hypertension and vascular dysfunction. Furthermore, it has shown protective effects in models of ischemia-reperfusion injury and cardiac hypertrophy.

Y-27632 has also been utilized in ophthalmological research, particularly in relation to glaucoma. By relaxing trabecular meshwork cells and increasing aqueous humor outflow, ROCK inhibition with Y-27632 has been associated with intraocular pressure reduction. This property has spurred the development of ROCK inhibitors as potential treatments for glaucoma.

In neurobiology, the compound has contributed to the understanding of ROCK's role in axon regeneration and neuronal survival. Inhibition of ROCK with Y-27632 has been shown to promote axonal growth and functional recovery in spinal cord injury models and to protect neurons in models of neurodegenerative diseases. These findings support the idea that modulation of the cytoskeleton through ROCK inhibition can influence neuronal plasticity and regeneration.

Y-27632 dihydrochloride is water-soluble and commonly used in cell culture experiments. It is typically applied at micromolar concentrations to exert its biological effects, and its action is reversible upon removal. While it is primarily a research tool, its influence on the development of therapeutic strategies targeting the ROCK pathway has been substantial.

The discovery and application of Y-27632 dihydrochloride have provided significant insights into the functional importance of the Rho/ROCK pathway. Its utility across a wide range of biological systems and disease models continues to make it a valuable compound in biomedical research.

References

2007. Inhibition of Rho-Kinase Impairs Fibroblast Stress Fiber Formation, Confluence, and Contractility In Vitro. Journal of Burn Care & Research, 28(3).
DOI: 10.1097/bcr.0b013e318053dad8

2013. Effects of Y27632 on Aqueous Humor Outflow Facility With Changes in Hydrodynamic Pattern and Morphology in Human Eyes. Investigative Ophthalmology & Visual Science, 54(8).
DOI: 10.1167/iovs.12-10930

2024. Y-27632 targeting ROCK1&2 modulates cell growth, fibrosis and epithelial-mesenchymal transition in hyperplastic prostate by inhibiting �-catenin pathway. Molecular Biomedicine, 5(1).
DOI: 10.1186/s43556-024-00216-9