| Size | Price | |
|---|---|---|
| 5mg | ||
| 10mg | ||
| Other Sizes |
| Targets |
ROCKII:12 nM (IC50); CaMKII:0.18 μM (IC50); PKG:0.36 μM (IC50); AuroraA:0.745 μM (IC50); MLCK:28.3 μM (IC50); EGFR:50 μM (IC50); MKK4:16.9 μM (IC50); GSK3α:60.7 μM (IC50); PKA:3.03 μM (IC50); Src:3.06 μM (IC50); PKC:5.68 μM (IC50); Abl:7.77 μM (IC50); AMPK:100 μM (IC50) P38α:100 μM (IC50)
|
|---|---|
| ln Vitro |
The IC50 for ROCK2 of the Rho kinase inhibitor H-1152 is 12 nM. With IC50 values of 0.180, 0.360, 0.745, respectively, for CaMKII, PKG, AuroraA, PKA, Src, PKC, MLCK, Abl, EGFR, MKK4, GSK3α, AMPK, and P38α, H-1152 (H-1152P) likewise exhibits reduced inhibitory action [1]. H-1152 has a 1.6 nM Ki for Rho kinase potency and a 0.63, 9.27, and 10.1 μM Ki for PKA, PKC, and MLCK, respectively, for PKA, PKC, and MLCK moderate inhibition. H-1152 (0.1-10 µM) exhibits a strong inhibitory effect on MARCKS phosphorylation in LPA-treated cells, with an IC50 value of 2.5 µM; however, no discernible effect is observed in PDBu-treated cells [2]. H-1152 (0.5-10 μM) does not result in a decrease in neuronal viability. Additionally, H-1152 (1, 5, or 10 μM) did not change the proportion of various neuronal morphologies. Moreover, in BMP4 and LIF cultures, H-1152 (10 μM) lengthens neurites [3].
|
| Enzyme Assay |
Protein kinase assay[2]
To determine the effect of inhibitors on Rho-kinase activity in vitro, inhibitors was added at the indicated concentrations to 50 µL of the assay mixture 50 mm Tris-HCl (pH 7.5), 5 mm MgCl2, 1 mm EDTA, 1 mm EGTA, 1 mm dithiothreitol, 40 µm S6-peptide, various concentrations of [γ-32P]ATP and purified Rho-kinase, which was purified as described previously (Nagumo et al. 2000). The reactions were started by the addition of [γ-32P]ATP and carried out at 30°C for 5 min. The Michaelis–Menten equation was used to calculate the Michaelis constant (Km) and maximal velocity (Vmax) of Rho-kinase. Data were further analyzed with secondary plot to calculate the inhibitory constant (Ki). The effects of Rho-kinase inhibitors on protein kinase A (PKA) and C (PKC) were assayed according to our method with minor modifications (Tokumitsu et al. 1991). The reaction for PKA was carried out at 37°C for 5 min in a 20-µL mixture containing 25 mm Tris-HCl (pH 7.0), 10 mm MgCl2, 2 mm EGTA, 50 µm peptide of cyclic AMP response element binding protein and 10 µm[γ-32P]ATP (150 cpm/pmol). The reaction for PKC was carried out for 5 min in 30 µL of a mixture that contained 50 mm Tris-HCl (pH 7.0), 10 mm MgCl2, 0.5 mm CaCl2, 0.2 mg/mL histone type III-S, 50 µg/mL phosphatidyl serine and 10 µm[γ-32P]ATP (150 cpm/pmol). The effect of Rho-kinase inhibitors on myosin light chain kinase (MLCK) was assayed under the conditions previously described elsewhere (Asano et al. 1989), in 30 µL of a reaction mixture that contained 40 mm HEPES (pH 7.0), 5 mm magnesium acetate, 0.5 mm CaCl2, 0.1% (v/v) Tween-80, 5.3 nm calmodulin, 51 µm myosin light chain peptide, 1.95 nm MLCK and [γ-32P]ATP (150 cpm/pmol)[2].
|
| Cell Assay |
ROCK inhibitor H-1152 was diluted in water and added in an additional 10 μl to cultures 24 h after plating. Water was added to controls. Eighteen hours after the addition of inhibitor, cultures were fixed in 4% paraformaldehyde (1 h at room temperature for peroxidase-linked labeling and 20 min at room temperature for fluorescence labeling). For ArrayScan/Cellomics automated analysis: Cells were plated in a total volume of 50 μl on 384 well plastic plates previously coated with poly-d-lysine/laminin, and cultured in the same medium[3].
To investigate the effects of the protein kinase inhibitors (e.g. H-1152 ), the cells were preincubated with the inhibitors for 15 min and then stimulated with LPA for 1 min or with PDBu for 5 min. At various determination points after the stimulation, 5% (v/v) trichloroacetic acid was added to the cultures and cells were scraped off with a rubber scraper. After sedimentation, the precipitate was washed with acetone containing 6 mm dithiothreitol to remove trichloro-acetic acid and then dried. The dried cell powder was placed in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) sample buffer containing 125 mm Tris (pH 7.4), 8 m urea, 10% (w/v) sucrose, 0.02% (w/v) bromophenol blue, 4% (w/v) SDS and 2-mercaptoethanol, and the preparation was then extracted by heating at 95°C for 5 min. The samples were passed through a 0.22-µm centrifugal filter (Millipore, Bedford, MA, USA) to remove indissoluble materials. The samples (2.5 × 105 cells) were subjected to SDS–PAGE (7.5%)/western blotting analysis. Proteins in the gel were electrotransferred to a polyvinylidene difluoride membrane. After incubation with 5% (w/v) skimmed milk, the membrane was blotted with pS159-Mar-Ab, followed by anti-mouse IgG antibody. Immunoreactivity was detected by enhanced chemiluminescence-western blotting detection reagents, and its extent was measured by scanning densitometry of the X-ray film using KODAK 1D Image Analysis Software, and expressed as a percentage versus control under each condition[2]. |
| References |
|
| Additional Infomation |
(S)-2-methyl-1-(4-methylisoquinoline-5-sulfonyl)-1,4-diazepane is a member of the class of isoquinolines that is the sulfonamide formed by the formal condensation of the sulfo group of 4-methylisoquinoline-5-sulfonic acid with the 1-amino group of (S)-2-methyl-1,4-diazepane. It has a role as an EC 2.7.11.1 (non-specific serine/threonine protein kinase) inhibitor. It is a member of isoquinolines and a N-sulfonyldiazepane. It is a conjugate base of a (S)-2-methyl-1-(4-methylisoquinoline-5-sulfonyl)-1,4-diazepane(2+).
|
| Molecular Formula |
C16H21N3O2S
|
|---|---|
| Molecular Weight |
319.423
|
| Exact Mass |
319.1354
|
| Elemental Analysis |
C, 60.16; H, 6.63; N, 13.16; O, 10.02; S, 10.04
|
| CAS # |
451462-58-1
|
| Related CAS # |
H-1152 dihydrochloride;871543-07-6;Glycyl H-1152 hydrochloride;913844-45-8
|
| PubChem CID |
448043
|
| Appearance |
Typically exists as Off-white to light yellow solids at room temperature
|
| LogP |
4.867
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
22
|
| Complexity |
476
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CC1=CN=CC2=C1C(S(=O)(N3[C@@H](C)CNCCC3)=O)=CC=C2
|
| InChi Key |
AWDORCFLUJZUQS-ZDUSSCGKSA-N
|
| InChi Code |
InChI=1S/C16H21N3O2S/c1-12-9-18-11-14-5-3-6-15(16(12)14)22(20,21)19-8-4-7-17-10-13(19)2/h3,5-6,9,11,13,17H,4,7-8,10H2,1-2H3/t13-/m0/s1
|
| Chemical Name |
4-Methyl-5-[[(2S)-2-methyl-1,4-diazepan-1-yl]sulfonyl]isoquinoline
|
| Synonyms |
H 1152; H1152; 451462-58-1; (S)-2-METHYL-1-[(4-METHYL-5-ISOQUINOLINE)SULFONYL]-HOMOPIPERAZINE; H-1152P; CHEBI:88220; 4-methyl-5-[[(2S)-2-methyl-1,4-diazepan-1-yl]sulfonyl]isoquinoline; H1152; (S)-4-Methyl-5-((2-methyl-1,4-diazepan-1-yl)sulfonyl)isoquinoline;
H-1152
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1307 mL | 15.6534 mL | 31.3067 mL | |
| 5 mM | 0.6261 mL | 3.1307 mL | 6.2613 mL | |
| 10 mM | 0.3131 mL | 1.5653 mL | 3.1307 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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