Y-27632 - ROCK 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEY-27632Cat. No.: HY-10071CAS No.: 146986-50-7分式: CHNO分量: 247.34作靶點: ROCK作通路: Cell Cycle/DNA Damage; Stem Cell/Wnt; TGF-beta/Smad儲存式: 4C, stored under nitrogen* In solvent : -80C, 6 months; -20C, 1 month (stored undernitrogen)溶解性

2、數(shù)據(jù)體外實驗 DMSO : 50 mg/mL (202.15 mM; Need ultrasonic)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 4.0430 mL 20.2151 mL 40.4302 mL5 mM 0.8086 mL 4.0430 mL 8.0860 mL10 mM 0.4043 mL 2.0215 mL 4.0430 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。體內(nèi)實驗 請根據(jù)您的實驗動物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢埾扰渲瞥吻宓膬湟海僖来翁砑又軇?為保證實

3、驗結(jié)果的可靠性，體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲備液可以根據(jù)儲存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (10.11 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (10.11 mM); Clear solution3. 請依序添加每種溶劑： 10% DMSO 90

4、% corn oilSolubility: 2.5 mg/mL (10.11 mM); Clear solution1/4 Master of Small Molecules 您邊的抑制劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 Y-27632種ATP競爭性的 ROCK-I 和 ROCK-II 抑制劑，Ki 分別為 220 nM 和 300 nM，Y-27632 通過上-間充質(zhì)過渡樣調(diào)節(jié)引發(fā)的誘導(dǎo)多能細(xì)胞 (hIPSCs) 選擇性地分化為間胚層譜系。IC50 & Target ROCK-I ROCK-II PKN Citron kinase220 nM (Ki)

5、300 nM (Ki) 3.1 M (Ki) 5.3 M (Ki)PKC PKA73 M (Ki) 25 M (Ki)體外研究 Y-27632 inhibits the ROCK family of kinases 100 times more potently than other kinases including proteinkinase C, cAMP-dependent kinase and myosin light chain kinase. Y-27632 prolongs the lag time and delaysthe appearance of BrdU-labele

6、d cells in a concentration-dependent manner, delays of about 1 and 4 h arenoticed in the Swiss 3T3 cells treated with 10 and 100 M Y-27632, respectively 1. Y-27632 promotesneuronal differentiation of adipose tissue-derived stem cells (ADSCs). Compared to 1.0 and 2.5 M Y-27632induced groups, percenta

7、ges of neuroal-like cells achieved a peak in the 5.0 M Y-27632 induced group 2.體內(nèi)研究 Y-27632 (5 and 10 mg/kg) significantly prolongs the onset time of myoclonic jerks when compare with salinegroup. Y-27632 (5 and 10 mg/kg) significantly prolongs the onset time of clonic convulsions when comparewith s

8、aline group 3. Treatment with Dimethylnitrosamine (DMN) causes a significant decrease in rat bodyand liver weight (DMN-S group) compared with control animals (S-S group). Oral Y27632 (30 mg/kg)essentially prevents this DMN-induced rat body and liver weight loss (DMN-Y group) 4.PROTOCOLKinase Assay 1

9、 Recombinant ROCK-I, ROCK-II, PKN, or citron kinase is expressed in HeLa cells as Myc-tagged proteins bytransfection using Lipofectamine, and is precipitated from the cell lysates by the use of 9E10 monoclonalanti-Myc antibody coupled to G protein-Sepharose. Recovered immunocomplexes are incubated w

10、ith variousconcentrations of 32PATP and 10 mg of histone type 2 as substrates in the absence or presence of variousconcentrations of either Y-27632 or Y-30141 at 30C for 30 min in a total volume of 30 L of the kinase buffercontaining 50 mM HEPES-NaOH, pH 7.4, 10 mM MgCl2, 5 mM MnCl2, 0.02% Briji 35,

11、 and 2 mMdithiothreitol. PKCa is incubated with 5 M 32PATP and 200 g/mL histone type 2 as substrates in theabsence or presence of various concentrations of either Y-27632 or Y-30141 at 30C for 10 min in a kinasebuffer containing 50 mM Tris-HCl, pH 7.5, 0.5 mM CaCl2, 5 mM magnesium acetate, 25 g/mL p

12、hosphatidylserine, 50 ng/mL 12-O-tetradecanoylphorbol-13-acetate and 0.001% leupeptin in a total volume of 30 L.Incubation is terminated by the addition of 10 L of 43 Laemmli sample buffer. After boiling for 5 min, themixture is subjected to SDS-polyacrylamide gel electrophoresis on a 16% gel. The g

13、el is stained withCoomassie Brilliant Blue, and then dried. The bands corresponding to histone type 2 are excised, and theradioactivity is measured 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 HeLa cells are plated at a density of 3104

14、 cells per 3.5-cm dish. The cells are cultured in DMEM containing10% FBS in the presence of 10 mM Thymidine for 16 h. After the cells are washed with DMEM containing2/4 Master of Small Molecules 您邊的抑制劑師www.MedChemE10% FBS, they are cultured for an additional 8 h, and then 40 ng/mL of Nocodazole is a

15、dded. After 11.5 h ofthe Nocodazole treatment, various concentrations of Y-27632 (0-300 M), Y-30141, or vehicle is added andthe cells are incubated for another 30 min 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 3Administration 34 Male,

16、 inbred Swiss albino mice (2-3 months old) weighing 25-30 g are used. Mice are injected with a sub-convulsive dose of PTZ (35 mg/kg, i.p.) (on Mondays, Wednesdays and Fridays) of each week for a total of11 injections. After each PTZ injection, mice are observed for 30 min and the occurrence of convu

17、lsiveactivity is recorded. After 30 min, the mice are then injected with either Fasudil (25 mg/kg, i.p.) or Y-27632 (5mg/kg, i.p.) and returned to their home cages until the next injection. Control mice for Fasudil and Y-27632receives saline.Rats 4Male Wistar Kind A rats (200-250 g) are used. DMN (1

18、 g/mL) is diluted ten times with saline (finalconcentration 1%) and 10 mg/kg per day of DMN is injected intraperitoneally (i.p.) on the first 3 days of eachweek for 4 weeks. Y27632 is given orally once per day at a dose of 30 mg/kg for 4 weeks starting on the dayof the first injection of DMN. The do

19、se of 30 mg/kg corrects hypertension in several rat models withouttoxicity. Twenty rats are randomized into four experimental groups (n=5 in each group) as follows: (1) S-S(injection of saline i.p. and oral administration of saline); (2) S-Y (injection of saline i.p. and oraladministration of Y27632

20、); (3) DMN-S (DMN i.p. and oral administration of saline); (4) DMN-Y (DMN i.p. andoral administration of Y27632). The rats are weighed every week. They are sacrificed at the end of the fourth week and the liver is excised. In addition, a blood sample is taken immediately before the rats are sacrific

21、ed.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Cell Rep. 2019 Jan 8;26(2):407-414.e5. Cancer Lett. 2019 Apr 28;448:117-127. Anal Chem. 2017 Oct 17;89(20):10841-10849. J Med Chem. 2019 Jun. J Exp Clin Cancer Res. 2018 Jun 28;37(1):128.See

22、 more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Ishizaki T, et al. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol Pharmacol. 2000May;57(5):976-83.2. Xue ZW, et al. Rho-associated coiled kinase inhibitor Y-27632 promotes neuronal-like differentiation of adult human adipose tissue-derived stem cells.Chin Med J (Engl). 2012 Sep;125(18):3332-5.3. Inan S, et al. Antiepileptic