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Y-27632 2HCl

Name: Y-27632 2HCl
Brand: Selleck Chemicals
SKU: S1049
Rating: 5 (1306 reviews)

Catalog No.S1049 Purity:99.89%

Y-27632 2HCl is a selective ROCK1 and ROCK2 inhibitor with a K_i 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

CAS No. 129830-38-2

Purity & Quality Control

Batch: Purity: 99.89%

99.89

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 PKP2^mut 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

Related Targets	ROCK1 ROCK2	Click to Expand
Related Products	Y-27632 Fasudil HCl Thiazovivin GSK429286A RKI-1447 Ripasudil hydrochloride dihydrate GSK269962A HCl Y-39983 Dihydrochloride Hydroxyfasudil HCl	Click to Expand
Related Compound Libraries	Kinase Inhibitor Library PI3K/Akt Inhibitor Library MAPK Inhibitor Library DNA Damage/DNA Repair compound Library Cell Cycle compound library	Click to Expand

Signaling Pathway

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

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 K_i 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
Kinase Assay	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. K_i value is calculated according to the equation, K_i = IC50/(1 + S/K_m), where S and K_m represent concentrations of and K_m 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)

References

+ Expand

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)	3 years-20°Cpowder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years-20°Cpowder		1 month-20°Cin solvent

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

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.				In vivo Formulation Calculator
Clear solution	saline	64.000mg/ml (199.84mM)	Taking the 1 mL working solution as an example, add 64 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	saline Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (31.22mM)	Taking the 1 mL working solution as an example, add 10 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Clear solution	saline	64.000mg/ml (199.84mM)	Taking the 1 mL working solution as an example, add 64 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	saline Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (31.22mM)	Taking the 1 mL working solution as an example, add 10 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Clear solution	saline	64.000mg/ml (199.84mM)	Taking the 1 mL working solution as an example, add 64 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	saline	64.000mg/ml (199.84mM)	Taking the 1 mL working solution as an example, add 64 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	saline	64.000mg/ml (199.84mM)	Taking the 1 mL working solution as an example, add 64 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

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Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

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

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

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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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