Y-27632 2HCl

Catalog No.S1049

Y-27632 2HCl Chemical Structure

Molecular Weight(MW): 320.26

Y-27632 2HCl is a selective ROCK1 (p160ROCK) inhibitor with Ki of 140 nM in a cell-free assay, exhibits >200-fold selectivity over other kinases, including PKC, cAMP-dependent protein kinase, MLCK and PAK.

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Cited by 81 Publications

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Biological Activity

Description Y-27632 2HCl is a selective ROCK1 (p160ROCK) inhibitor with Ki of 140 nM 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

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]
Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Swiss 3T3 cells M37RT2Z2dmO2aX;uJGF{e2G7 M3ezSlExKM7:TR?= MmS3NkBp NUK5T4pMTE2VTx?= NVvQd29FUW6qaXLpeJMhfGinIHHzd4Vu[my7IH;mJI1q[3KxdIXieYxmeyCjbnSgbY51\XKvZXTpZZRmKG[rbHHt[Y51eyC2bzDmc5JuKGW6dHXu[IVlKHC{b3Pld5Nmew>? MX25OlQ4PjV2
N1E-115 M3PROWZ2dmO2aX;uJGF{e2G7 NUjGPVlPOTBizszN M2DjVFIhcA>? MU\EUXNQ NXPCeGo{UW6qaXLpeJMhfGinIHHzd4Vu[my7IH;mJI1q[3KxdIXieYxmeyCjbnSgbY51\XKvZXTpZZRmKG[rbHHt[Y51eyC2bzDmc5JuKGW6dHXu[IVlKHC{b3Pld5Nmew>? MmrxPVY1PzZ3NB?=
HeLa NIjDPWxHfW6ldHnvckBCe3OjeR?= Mn:3NVAh|ryP NF7hSXU{OCCvaX6= NF\tSIlKdmirYnn0d{B1cGViZn;ycYF1cW:wIH;mJJN1emW|czDmbYJmenNiYX7kJJRp\SCjc4PlcYJtgSCxZjD2bY5kfWyrbj3jc451[WmwaX7nJIZw[2GuIHHkbIV{cW:wcx?= M2nnSlk3PjhyN{K=
CCL39 M1jnN2Z2dmO2aX;uJGF{e2G7 MnzFN|Ah|ryP Mm\pN|AhdWmw NGPUWZFEd22ybHX0[Yx6KGGkb3zpd4hmeyCjY4TpeoF1cW:wIH;mJG5iNUhiZYjjbIFv\2W{IF7ISVEh[nliaX70[YdzcW6| NETsSJQ6Pjl|M{iy
Mesothelial cells from rat mesentery M2TZWmlvfmG|aY\lJGF{e2G7 NWLIV5N3OzBizszN MUGyNEBp Mn7jRoxw[2u|IHnueoF{cX[nIHHjeIl3cXS7 NYXDOI1tQTl|MEi3Ni=>
NIH3T3 M3m5d2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NX\4VXhtOTBizszN NVjIeHhCOThiZB?= NYTW[FlbTG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? NV:3cZFHOTByMkGzPFY>
Dbl-d MWrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? Mnv0NVAh|ryP NIe2NG8yQCCm MnjKV5Rzd26pbImgbY5pcWKrdIOgZ4VtdCCpcn;3eIg> MkjDNVAxOjF|OE[=
Dbl-e NWXrUpRLT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M{S0OFExKM7:TR?= MUmxPEBl NW[2UnNDVW:mZYLheIVtgSCrbnjpZol1eyClZXzsJIdzd3e2aB?= NF;tNmoyODB{MUO4Oi=>
mNET1-d M4\KS2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NV7J[WU3OTBizszN Mlf2NVgh\A>? NUiwOnpiW3S{b37ncJkhcW6qaXLpeJMh[2WubDDndo94fGh? Mk\KNVAxOjF|OE[=
mNET1-e MlLxS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M3PpdVExKM7:TR?= MWKxPEBl MXfTeJJwdmeueTDpcohq[mm2czDj[YxtKGe{b4f0bC=> NWDIR|dROTByMkGzPFY>
Ras-2 MV;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MWmxNEDPxE1? MXSxPEBl MXHTeJJwdmeueTDpcohq[mm2czDj[YxtKGe{b4f0bC=> M{nUTVExODJzM{i2
Ras-4 MWTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MVyxNEDPxE1? NXLHZYdoOThiZB?= NYO2U|UzW3S{b37ncJkhcW6qaXLpeJMh[2WubDDndo94fGh? NIjaR4syODB{MUO4Oi=>
Src-1 NGPLPG1Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M{mzXVExKM7:TR?= MoTWNVgh\A>? NH\ab29Fd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp MUKxNFAzOTN6Nh?=
Src-4 MV;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NWnwVWdROTBizszN NX3ZdG8xOThiZB?= MXnEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To NGr5SIUyODB{MUO4Oi=>
NIH3T3 NVfPc45JT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Mn3nNVAh|ryP NUHadZVTOThiZB?= NX21XIZ3TG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? NYnLXpBjOTByMkGzPFY>
Src-1 MofMS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NVvtdVFMOTBizszN M3\ZcVE5KGR? NVTvZ3JoTG:nczDuc5QhcW6qaXLpeEBk\WyuIHfyc5d1cA>? NEfRRXQyODB{MUO4Oi=>
Src-2 MXvHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NHXlbJoyOCEQvF2= M1fP[lE5KGR? NHvJfmJFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp NImzWmYyODB{MUO4Oi=>
SW620 NHvkcolIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MUGxNEDPxE1? NGLUNpMyQCCm NGDtcpFFd2W|IH7veEBqdmirYnn0JINmdGxiZ4Lve5Rp M33XRlExODJzM{i2
HCT15 MYXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M1H6XVExKM7:TR?= MYqxPEBl MXPEc4V{KG6xdDDpcohq[mm2IHPlcIwh\3Kxd4To MkDuNVAxOjF|OE[=
HCT116 NWTnTlZLT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Ml\ENVAh|ryP MYmxPEBl M{nWcnN1em:wZ3z5JIlvcGmkaYTzJINmdGxiZ4Lve5Rp M3GwTFExODJzM{i2
LS174T NV[1TY9yT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NV;ue4RTOTBizszN MnPmNVgh\A>? M1\kbG1w\GW{YYTlcJkhcW6qaXLpeJMh[2WubDDndo94fGh? MYqxNFAzOTN6Nh?=
Neonatal rat ventricular myocytes NELWVmFHfW6ldHnvckBCe3OjeR?= NFracXYyOCEQvF2= M{XyTlQ5KGh? MYHJcohq[mm2czDFWE0yNWmwZIXj[YQhcW6lcnXhd4V{KGmwIIDyc5RmcW5ic4nueIhme2m|LDDj[YxtKHOrenWgZY5lKG27b3\pZpJqdGyjcjDvdodidmm8YYTpc44> NFvpN2syODN6Nk[xNy=>
Stellate Cell M1jNVWZ2dmO2aX;uJGF{e2G7 NIPyelkzPSEQvF2= NHfX[pYyPSCvaX6= NYnoVpdxUW6qaXLpeJMh\m:{bXH0bY9vKG:oIF[tZYN1cW5ic4Ty[ZN{KG[rYnXyd{BidmRicHjvd5Bpd3K7bHH0bY9vKG:oIH35c5NqdiCuaXfoeEBkcGGrbh?= MVexNFYxODR7Nh?=
Rat Vascular Smooth Muscle Cells MX\GeY5kfGmxbjDBd5NigQ>? NXTyfmxXOTBizszN NXjZWnF4OiCq MWfJcohq[mm2czDhcodqd3SnboPpckBKUS2rbnT1Z4VlKGi7cHXyeJJweGi7 MXqxNFY1OjNzNx?=
PC3 MVLGeY5kfGmxbjDBd5NigQ>? MXmyOUDPxE1? MlOwNUBp MlftTY5lfWOnczDtc5JxcG:ub3fpZ4FtKGOqYX7n[ZM> MlfINVA4OjB2N{G=
PC3 MmHVUYloemG2aX;uJGF{e2G7 MWmyOUDPxE1? M3;mdVEhcA>? MnmxTY5pcWKrdIOgeIhmKEKPRlKtR20h[W6mIITo[UBGT0Zvc4TpcZVt[XSnZDDtbYdz[XSrb36= M1r4OVExPzJyNEex
PC3 NEHobVFIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NYnVXWx5OjVizszN NIfGVmsyPyCq MnPZSI9meyCwb4SgbY5pcWKrdDDj[YxtKGe{b4f0bC=> NUW4cm83OTB5MkC0O|E>
LNCaP NYrIUIZIT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MV6yOUDPxE1? Mn7tNVchcA>? M1;VVGRw\XNibn;0JIlvcGmkaYSgZ4VtdCCpcn;3eIg> NXnLUVN{OTB5MkC0O|E>
Rat hepatic stellate cells NHPkUmtHfW6ldHnvckBCe3OjeR?= NVq5RlNxOzBizszN MVu0PEBp MWLEbY1qdmm|aHXzJJRp\SCyaH;zdIhwenmuYYTpc44hd2ZiRYLrNkwh[W6mIHTlZ5Jm[XOnczDu[ZchTE6DIIP5cpRp\XOrcx?= MmfJNVA5PDV4NkO=
Pancreatic acinar cells MWfGeY5kfGmxbjDBd5NigQ>? NF\uUVQyOMLizszN M2rnXlcxKG2rbh?= NXPweYtJWG:2ZX70bYF1\XNiQ1PLMZN1cW23bHH0[YQheGGwY4LlZZRq[yCnbor5cYUhe2WlcnX0bY9v NVi1V29SOTJ5NEWwPFA>
C2C12 Mm\rSpVv[3Srb36gRZN{[Xl? MVWxNOKh|ryP M4f4UVYhcA>? NIPLdYFRemW4ZX70d{B1cGVic3XybY5mKHCqb4PwbI9zgWyjdHnvckBw\iCLUmOtNUBqdmS3Y3XkJIJ6KGmwc4XsbY4h[W6mL3;yJHRPTi4QsR?= M3SzVFE3OjZ5MUK0
PC 12 MVvGeY5kfGmxbjDBd5NigQ>? M4rVSlExyqEQvF2= M4\ydVI1KGh? NHm4dYJCfHSnboXheIV{KGOjdHXjbI9t[W2rbnWgZolwe3mwdHjld4l{ MVKxOlIyQTR{NB?=
Cynomolgus monkey embryonic stem cells NXrONJRiS3m2b4TvfIlkKEG|c3H5 NGLXXYIzOCEEtV2= MlHJNlQhcA>? MVLQdo9ud3SnczDjfWVUKGOnbHygd5Vzfmm4YXy= Mn:3NVg6PDB6NUW=
TSGH 8301 NGjjNY1OcWe{YYTpc44hSXO|YYm= NHOzR2wzOCEEtV2= NYj2XGw6OSCq MoP5TY5kemWjc3XzJINmdGxibXnndoF1cW:w MWixPVg6PjR5NR?=
Swiss3T3 M{TFTWNwdG:weT3mc5JucW6pIFHzd4F6 NEfWOGoyOCEEtV2= MW[xN{Bl M4fuXWlv[3KnYYPld{Bxem:|dHH0[UBk\WyuIHPvcI9vgS2ob4LtbY5oKGGldHn2bZR6 NUTpSY5NOjF2NkS5NFI>
HT22 NFj2XZdEgXSxdH;4bYMhSXO|YYm= M4DI[FExKML3TR?= NIrOSFMyOyCq NY[5[YJSWHKxdHXjeJMh[WejaX7zeEBodHW2YX3heIUucW6mdXPl[EBv\XW{b37hcEBl\WG2aB?= M1fITVIzQDFyOEO1
Salivary gland stem cells M1HtXWZ2dmO2aX;uJGF{e2G7 M4jSTlExKML3TR?= MnGxO{Bl NYfLO2lGWmWmdXPld{BUT1OFIIPlcoV{[2WwY3W= NXfFNWMzOjV6MES1OlA>

... Click to View More Cell Line Experimental Data
Assay
Methods Test Index PMID
Western Blot
p-LIMK1(Thr508); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

p-LIMK2(Thr505); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

p-Cofilin(Ser3); 

PubMed: 24704720     


Immunoblot analysis of HCT116 and HCT116 SMAD4-/- cells. Cells were transfected transiently with either BMPR2 or pcDNA vector. Subsequently, cells were treated with 2.5 μmol/L of ROCK inhibitor (Y-27632) or PBS. Blots were incubated with antibodies against pLIMK1/2 and p-cofilin. Actin was used as a loading control.

CDK2; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

KRT7; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

ROCK2; 

PubMed: 26555939     


Western blot detected proteins changes pTR cells cultured with Y-27632. CDX2 was remarkably increased in Y-27632 treated pIVFTR-7 and pPATR-5 cells. pTR cells cultured without Y-27632 in the same condition are used as control. β-ACTIN serves as an endogenous control. 

E-cadherin; 

PubMed: 27399334     


The expression and/or phosphorylation of E-cadherin and ROCKs downstream targets in SW-480-FOXM1D cells treated with Y-27632.

p-MLC2(Ser19); 

PubMed: 27399334     


The expression and/or phosphorylation of E-cadherin and ROCKs downstream targets in SW-480-FOXM1D cells treated with Y-27632.

24704720 26555939 27399334
Transwell migration assay
cell migration inhibition ; 

PubMed: 27694793     


Data represent mean ± SEM, n=3. Compared with control, * P<0.05, ** P<0.01. Compared with control, Y-27632, or EGCG, *** P<0.01.

27694793
Immunofluorescence
Neurotoxicity Assay; 

PubMed: 21362567     


Representations of neurodegeneration in H9-, HUF5- and G2019S-iPSC derived neurons when treated with neurotoxins, H2O2 and MG-132, and ROCK inhibitor Y-27632. Scale bar = 50 µm.

E-cadherin; 

PubMed: 24523903     


The localized patterns of E-cadherin and b-catenin were examined using specific antibodies through confocal laser microscopy. MCF-7 cells were cultured for 24 hrs in the absence or presence of Y-27632 (20 µM) to inhibit ROCK activity. 

beta-catenin; 

PubMed: 24523903     


The localized patterns of E-cadherin and b-catenin were examined using specific antibodies through confocal laser microscopy. MCF-7 cells were cultured for 24 hrs in the absence or presence of Y-27632 (20 µM) to inhibit ROCK activity. 

Phalloidin; 

PubMed: 27399334     


Triple IF staining for F-actin (phalloidin, green) E-cadherin (b) (red) and nuclei (DAPI, blue) showed decreased F-actin, increased E-cadherin and altered cell shape and size in SW-480-FOXM1D cells treated with the ROCKs inhibitor Y-27632 or fasudil (both 10 μM, for 24 h). Scale bar, 25 μm. 

21362567 24523903 27399334
Growth inhibition assay
cell proliferation ; 

PubMed: 24523903     


MCF-7 cells and MDA-MB-231 cells were cultured on tissue culture plates for four days with or without Y-27632 (20 µM). Relative cell proliferation rates were determined by the cell number assayed using CCK-8 kit. The data are expressed as the mean ± SD. n = 3 culture dishes. The p-value was less than 0.05 for comparisons between control (Control) and treatment groups (Y-27632) in MCF-7 cells. *, p<0.05.

24523903
ELISA
caspase-9; 

PubMed: 30320378     


Caspase-9 activity was measured via ELISA. Mst1-mediated casapse-9 activation was inhibited by Y-27632 in A549 cells. *P<0.05. Ad, adenovirus; Mst1, mammalian STE20-like kinase 1.

30320378
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]

Protocol

Animal Research:[1] [7]
+ Expand
  • Animal Models: Male Wistar rats with spontaneous or induced hypertension; Swiss albino mice with Ehrlich ascites carcinoma
  • Formulation: Dissolved in DMSO, and diluted in saline (Rat); 0.9% NaCl (Mice)
  • Dosages: 30 mg/kg/day (Rat); 0-10 mg/kg (mice)
  • Administration: Orally (Rat); i.p. (Mice)
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 64 mg/mL warmed (199.83 mM)
Water 14 mg/mL (43.71 mM)
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
saline
For best results, use promptly after mixing. 		10mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 320.26
Formula

C14H21N3O.2HCl
CAS No. 129830-38-2
Storage powder
in solvent
Synonyms N/A

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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

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

  • * Indicates a Required Field

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.

selleck catalog

ROCK Signaling Pathway Map

ROCK Inhibitors with Unique Features

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  • Most Potent ROCK Inhibitor

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  • ROCK Inhibitor in Clinical Trial

    Fasudil (HA-1077) HCl : Phase III for scleroderma.

  • Newest ROCK Inhibitor

    RKI-1447 : Highly potent inhibitor of ROCK1 and ROCK2, with IC50 of 14.5 nM and 6.2 nM, respectively.

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    Features:Greater selectivity than Y-27632.

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    RKI-1447 is a potent inhibitor of ROCK1 and ROCK2, with IC50 of 14.5 nM and 6.2 nM, respectively, has anti-invasive and antitumor activities.

  • Palbociclib (PD-0332991) HCl

    Palbociclib (PD-0332991) HCl is a highly selective inhibitor of CDK4/6 with IC50 of 11 nM/16 nM in cell-free assays, respectively. It shows no activity against CDK1/2/5, EGFR, FGFR, PDGFR, InsR, etc. Phase 3.

    Features:Non-cytotoxic, and halts cancer cell growth & could be used in glioblastoma that has relapsed after temozolomide treatment (a chemotherapeutic used to treat many cancers).

  • Alisertib (MLN8237)

    Alisertib (MLN8237) is a selective Aurora A inhibitor with IC50 of 1.2 nM in a cell-free assay. It has >200-fold higher selectivity for Aurora A than Aurora B. Phase 3.

    Features:First orally available inhibitor of Aurora A.

  • Adavosertib （MK-1775）

    MK-1775 is a potent and selective Wee1 inhibitor with IC50 of 5.2 nM in a cell-free assay; hinders G2 DNA damage checkpoint. Phase 2.

    Features:The first reported Wee1 inhibitor.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID