Y-27632 2, inhibitor

Y-27632 dihydrochloride

Cat. Num. AAA14580

• Purity: 99.66%
• Synonyms: Y-27632 2; N/A; Y-27632 dihydrochloride; Y-27632 2, inhibitor

• Purity/Purification: 99.66%

• Molecular Formula: C14H23Cl2N3O
• CAS No.: 129830-38-2
• Appearance: Solid
• Solubility: DMSO: 199.8 mM; Water: 43.7 mM; (< 1 mg/ml refers to the product slightly soluble or insoluble)
• HNMR: Consistent with structure
• HPLC: Consistent with structure
• Conclusion: The test result is consistent with structure.
• In vitro Activity: The inhibited potency of Y-27632 against the ROCK family is 100 times than other kinases including protein kinase C, cAMP-dependent kinase and myosin light chain kinase. Y-27632 prolongs the lag time and delays the appearance of BrdU-labeledcells in a concentration-dependent manner, delays of about 1 and 4 h are noticed in the Swiss 3T3 cells treated with 10 and 100uM Y-27632, respectively [1]. The application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cellsmarkedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from approximately 1% to approximately27%) and facilitates subcloning after gene transfer. Furthermore, dissociated hES cells treated with Y-27632 are protected fromapoptosis even in serum-free suspension (SFEB) culture and form floating aggregates [2]. Y-27632 promotes neuronaldifferentiation of adipose tissue-derived stem cells (ADSCs). Compared to 1.0 and 2.5 uM Y-27632 induced groups,percentages of neuronal-like cells achieved a peak in the 5.0 uM Y-27632 induced group [3]. Y-27632 selectively inhibitssmooth-muscle contraction by inhibiting Ca2+ sensitization. We identified the Y-27632 target as a Rho-associated proteinkinase, p160ROCK. Y-27632 consistently suppresses Rho-induced [4]
• In vivo Activity: Y-27632 significantly decreased the blood pressure in a dose-dependent manner in spontaneously hypertensive rats: a fall of 50mm Hg was still observed 7 h after administration of 30 mg/kg of Y-27632. The same dose of this compound also caused asignificant and persistent fall in blood pressure in renal hypertensive rats, as well as in deoxycorticosterone acetate (DOCA)-salthypertensive rats. On the other hand, administration of the same dose of Y-27632 caused only a slight and transient fall in bloodpressure in control Wistar rats [4]. Y-27632 (5-10 mg/kg) and fasudil 5-25 (mg/kg) diminished onset of myoclonic jerks, clonicconvulsions and tonic hindlimb extensions in mice given pentylenetetrazole [5].
• Kinase Assay: Recombinant ROCK1/2, PKN or citron kinase is expressed in Hela cells as Myc-tagged proteins by transfection using Lipofectamine and is precipitated from the ceillysates by the use of 9E10 monoclonal anti-Myc antibody coupled to G protein-Sepharose. Recovered immunocomplexes are incubated with various concentrations of [32P]ATP and 10 mg of histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632 or Y-30141 at 30°C for 30 min in a total volume of 30 ul of the kinase buffer containing 50 mM HEPES-NaOH, pH 7.4, 10 mM MgCI2, 5 mM MnCI2, 0.02% Briji 35, and 2 mM dithiothreitol. PKCa is incubated with 5 uM [32P]ATP and 200 ug/ml histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632 or Y-30141 at 30°C for 10 min in a kinase buffer containing 50 mM Tris-HCI, pH 7.5, 0.5 mM CaCl2, 5 mM magnesium acetate, 25 ug/ml phosphatidylserine, 50 ng/ml 12-0-tetradecanoyl phorbol-13-acetate and 0.001 % leupeptin in a total volume of 30 uL. Incubation is terminated by the addition of 10 uL of 43 laemmli sample buffer. After boiling for 5 min, the mixture is subjected to SDS-polyacrylamide gel electrophoresis on a 16% gel. The gel is stained with Coomassie Brilliant Blue and then dried. The bands corresponding to histone type 2 are excised, and the radioactivity is measured.
• Cell Assay: Cell AssayHeLa cells are plated at a density of 3×10^4 cells per 3.5-cm dish. The cells are cultured in DMEM containing 10% FBS in thepresence of 10 mM Thymidine for 16 h. After the cells are washed with DMEM containing 10% FBS, they are cultured for anadditional 8 h, and then 40 ng/mL of Nocodazole is added. After 11.5 h of the Nocodazole treatment, various concentrations ofY-27632 (0-300 uM) or vehicle is added and the cells are incubated for another 30 min [1]
• Animal Research: A group of animals was injected with a single dose of pentylenetetrazole (PTZ, 65?mg/kg) to investigate if the two Rho-kinase inhibitors, fasudil, and Y-27632, changed the onset of PTZ seizures. Fasudil, Y-27632 or saline was given intraperitoneally 30?min before the PTZ injection. Each mouse was then observed for a 15-min period to measure the onset of the first myoclonic jerk, the onset of the first clonic convulsion and the occurrence of tonic hindlimb extension. Some of the animals died after tonic hindlimb extension, which is an expected outcome of acute PTZ injection. After the observation period, all animals were killed by halothane anesthesia [5]. Seven-week-old male Wistar rats were anesthetized with sodium pentobarbital. A silver clip (0.2 mm in diameter) was placed on the left renal artery in the preparation of the renal hypertensive rats. In the preparation of the DOCA-salt hypertensive rats, the left kidney was removed and a DOCA pellet (50 mg) was implanted subcutaneously. The DOCA rats were then fed an 8% salt diet. Rats from both groups were used after 8 weeks in the experiments, together with a male, 17–22-week old spontaneously hypertensive rats. The average systolic pressure in these groups of hypertensive rats ranged from 209 to 237 mm Hg, and no significant difference was found between groups. Eight-week-old male Wistar rats were used as controls. Their average systolic pressure was 139 mm Hg. Y-27632was administered orally. The systolic blood pressure was measured by the tail cuff method at 1, 3, 5, 7 and 24 h. The rats were prewarmed to 40 8C for 10 min before each measurement. No toxicity was found in rats treated with 30 mg kg?1 of Y-27632 administered per os once per day for 10 days [4].
• Preparation and Storage: 0-4°C For short term (days to weeks), or -20°C for long term (months)

Structure

Related Product Information for Y-27632 2, inhibitor

Y-27632 is a selective inhibitor of ROCKs including p160ROCK (Ki: 140 nM) and ROCK2 (IC50: 800 nM).