Home Cell Cycle ROCK inhibitor - Autophagy activator Y-27632 2HCl For research use only.

Y-27632 2HCl

Y-27632 2HCl is a selective ROCK1 and ROCK2 inhibitor with a Ki of 140 nM and 300nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.

Y-27632 2HCl Chemical Structure

Y-27632 2HCl Chemical Structure

CAS No. 129830-38-2

Purity & Quality Control

Products often used together with Y-27632 2HCl

SB202190

Y-27632 2HCl and SB202190, along with other small molecule inhibitors, generate inducible mesenchymal stromal/stem cells (MSCs)-like cells (iMSCs) from human primary dermal fibroblasts.

Lai PL, et al. Sci Rep. 2017 Mar 17;7:44534.

SB431542

Y-27632 2HCl and SB431542 combination reverses the mesenchymal actin cytoskeleton stress fibers induced by TGF-β1 in TGF-β1 knockout mice cells (mTEC-KO).

Das S, et al. BMC Cell Biol. 2009 Dec 21;10:94.

Laduviglusib (CHIR-99021)

Y-27632 2HCl and CHIR99021 collectively reduce the expression of adipogenic genes in both wild-type (WT) and PKP2mut cardiac myocytes (CMs).

Parrotta EI, et al. Int J Mol Sci. 2021 Feb 18;22(4):2004.

MG132

Y-27632 reduces P-IκBα polyubiquitination caused by TNFα with MG132 pre-treatment in MYLA cells.

Gallardo F, et al. Leukemia. 2018 Oct;32(10):2211-2223.

Z-VAD-FMK

Y27632 2HCl and Z-VAD-FMK block membrane blebbing and nucleus condensation induced by H2O2 in PC12 cells.

Wang Y, et al. Front Mol Neurosci. 2017 Mar 14;10:75.

Y-27632 2HCl Related Products

Signaling Pathway

ROCK Signaling Pathways

ROCK Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Salivary gland stem cells Function Assay 10 µM 7 d Reduces SGSC senescence 25804560
HT22 Cytotoxic Assay 10 µM 13 h Protects against glutamate-induced neuronal death 22810835
Swiss3T3 Colony-forming Assay 10 µM 13 d Increases prostate cell colony-forming activity 21464902
TSGH 8301 Migration Assay 20 µM 1 h Increases cell migration 19896475
Cynomolgus monkey embryonic stem cells Cytotoxic Assay 20 µM 24 h Promotes cyES cell survival 18940855
PC 12 Function Assay 10 μM 24 h Attenuates catecholamine biosynthesis 16219424
C2C12 Function Assay 10 μM 6 h Prevents the serine phosphorylation of IRS-1 induced by insulin and/or TNF-α 16267124
Pancreatic acinar cells Function Assay 10 μM 70 min Potentiates CCK-stimulated pancreatic enzyme secretion 12745080
Rat hepatic stellate cells Function Assay 30 μM 48 h Diminishes the phosphorylation of Erk2, and decreases new DNA synthesis 10845663
LNCaP Growth Inhibition Assay 25 μM 17 h Does not inhibit cell growth 10720471
PC3 Growth Inhibition Assay 25 μM 17 h Does not inhibit cell growth 10720471
PC3 Migration Assay 25 μM 1 h Inhibits the BMFB-CM and the EGF-stimulated migration 10720471
PC3 Function Assay 25 μM 1 h Induces morphological changes 10720471
Rat Vascular Smooth Muscle Cells Function Assay 10 μM 2 h Inhibits angiotensin II-induced hypertrophy 10642317
Stellate Cell Function Assay 25 μM 15 min Inhibits formation of F-actin stress fibers and phosphorylation of myosin light chain 10600496
Neonatal rat ventricular myocytes Function Assay 10 μM 48 h Inhibits ET-1-induced increases in protein synthesis, cell size and myofibrillar organization 10386613
LS174T Growth Inhibition Assay 10 μM 18 d Moderately inhibits cell growth 10021386
HCT116 Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
HCT15 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
SW620 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Src-2 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Src-1 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
NIH3T3 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Src-4 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Src-1 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Ras-4 Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
Ras-2 Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
mNET1-e Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
mNET1-d Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
Dbl-e Growth Inhibition Assay 10 μM 18 d Moderately inhibits cell growth 10021386
Dbl-d Growth Inhibition Assay 10 μM 18 d Strongly inhibits cell growth 10021386
NIH3T3 Growth Inhibition Assay 10 μM 18 d Does not inhibit cell growth 10021386
Mesothelial cells from rat mesentery Invasive Assay 30 μM 20 h Blocks invasive activity 9930872
CCL39 Function Assay 30 μM 30 min Completely abolishes activation of Na-H exchanger NHE1 by integrins 9693382
HeLa Function Assay 10 μM 30 min Inhibits the formation of stress fibers and the assembly of vinculin-containing focal adhesions 9668072
N1E-115 Function Assay 10 μM 2 h DMSO Inhibits the assembly of microtubules and intermediate filaments to form extended processes 9647654
Swiss 3T3 cells Function Assay 10 μM 2 h DMSO Inhibits the assembly of microtubules and intermediate filaments to form extended processes 9647654
Click to View More Cell Line Experimental Data

Biological Activity

Description Y-27632 2HCl is a selective ROCK1 and ROCK2 inhibitor with a Ki of 140 nM and 300nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.
Targets
ROCK1 (p160ROCK) [1]
(Cell-free assay)		 ROCK2 [6]
(Cell-free assay)
140 nM(Ki) 300 nM(Ki)
In vitro
In vitro

Y-27632 2HCl inhibits ROCK-II while displaying little activity against PKC, cAMP-dependent protein kinase and myosin light-chain kinase (MLCK) with Ki of 26 μM, 25 μM and > 250 μM, respectively, as well as PKA activated by another Rho-family GTPase member, Cdc42. Y-27632 2HCl inhibits smooth-muscle contraction induces by various agonists including phenylephrine, histamine, acetylcholine, serotonin, endothelin, and thromboxane with IC50 of 0.3-1 μM, by selectively inhibiting Ca2+ sensitization. Y-27632 2HCl suppresses Rho-induced, p160ROCK-mediated formation of stress fibres in cultured cells. [1]

Y-27632 2HCl treatment blocks both Rho-mediated activation of actomyosin and LPA-stimulated invasive activity of MM1 cells in a concentration-dependent manner. [2]

Y-27632 2HCl treatment is not only sufficient to initiate formation of exuberant axonal processes but also facilitates axonal maturation during the very early stages of axonogenesis, while largely sparing axon elongation. [3]

In human embryonic stem (hES) cells, Y-27632 2HCl treatment at 10 μM markedly diminishes dissociation-induced apoptosis even in serum-free suspension (SFEB) culture, increases cloning efficiency (from ~1% to ~27%), facilitates subcloning after gene transfer, and enables SFEB-cultured hES cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors. [4]
Kinase Assay Phosphorylation reactions
The p160ROCK is expressed in COS cells as tagged full-length proteins, and immunoprecipitated by the use of anti-tag antibodies. The p160ROCK (30 ng) is incubated with 40 μM [γ-32P]ATP (3.3 Ci/mmol) and with 3 μg of either histone (HF2A), dephosphorylated casein or MBP in the presence of various concentrations of Y-27632 2HCl at 30 °C in a total volume of 31 μL. A 7 μL aliquot is taken at 0, 5, 10, and 20 minutes, mixed with an equal volume of 2 × Laemmli sample buffer, and applied to SDS-PAGE. The gel is stained with Commassie Blue, dried and subjected to analysis by a Bioimage Analyzer BAS2000. The Y-27632 2HCl concentration required to inhibit p160ROCK activity by 50% (IC50 value) is obtained. Ki value is calculated according to the equation, Ki = IC50/(1 + S/Km), where S and Km represent concentrations of and Km value for ATP.
Cell Research Cell lines hES Cells
Concentrations 10 uM
Incubation Time 1 h
Method

Y-27632 was added to culture medium at 10 uM 1 h before detaching the cells from the feeder layer and also upon seeding the cells onto a new MEF layer.
Experimental Result Images Methods Biomarkers Images PMID
Western Blot p-LIMK1(Thr508) p-LIMK2(Thr505) p-Cofilin(Ser3) CDK2 KRT7 ROCK2 E-cadherin p-MLC2(Ser19) 24704720
Transwell migration assay cell migration inhibition 27694793
Immunofluorescence Neurotoxicity Assay E-cadherin beta-catenin Phalloidin 21362567
Growth inhibition assay cell proliferation 24523903
ELISA caspase-9 30320378
In Vivo
In vivo

Oral administration of Y-27632 2HCl at 30 mg/kg significantly decreases the blood pressure in a dose-dependent manner in spontaneous hypertensive rats, renal hypertensive rats, as well as deoxycorticosterone acetate (DOCA)-salt hypertensive rats. [1]

When Y-27632 2HCl is continuously administered at a rate of 0.55 μL per hour by implanted pumps for 11 days tumor cell invasion (MM1 cells expressing Val14-RhoA in rats) is significantly delayed. [2]

By inhibiting ROCK, Y-27632 2HCl treatment attenuates hypoxia-induced angiogenesis and vascular remodeling in the pulmonary circulation. [5] Pretreatment with Y-27632 has a protective effect against tumor formation in albino mice with Ehrlich ascites carcinoma. [7]
Animal Research Animal Models Male Wistar rats with spontaneous or induced hypertension; Swiss albino mice with Ehrlich ascites carcinoma
Dosages 30 mg/kg/day (Rat); 0-10 mg/kg (mice)
Administration Orally (Rat); i.p. (Mice)

Chemical Information & Solubility

Molecular Weight 320.26 Formula

C14H21N3O.2HCl
CAS No. 129830-38-2 SDF Download Y-27632 2HCl SDF
Smiles CC(C1CCC(CC1)C(=O)NC2=CC=NC=C2)N.Cl.Cl
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 64 mg/mL ( (199.83 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : 64 mg/mL

Ethanol : 4 mg/mL


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
Is there any data about the Amax (maximum attraction luminosity) and extinction coefficient of this compound?

Answer:
The wavelength we used to test HPLC is 260nm while the extinction coefficient is unknown.

Question 2:
Could this product be used in cell culture? Do you have any reference for this application?

Answer:
Yes. The Y-27632 can be used in cell culture certainly. Here is the reference website: http://molpharm.aspetjournals.org/content/57/5/976.full.

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