| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
ROCK1 14.5 nM (IC50) ROCK2 6.2 nM (IC50) Apoptosis
RKI-1447 targets Rho-associated protein kinase (ROCK) 1 and 2, which are serine/threonine kinases that act as downstream effectors of the small GTPase RhoA. ROCK kinases play critical roles in the regulation of actin cytoskeleton organization, cell contraction, adhesion, migration, and gene expression. By inhibiting ROCK activity, RKI-1447 disrupts these cellular processes and modulates various physiological and pathological functions. |
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| ln Vitro |
In human cancer cells, RKI 1447 inhibits the phosphorylation of the ROCK substrates MLC-2 and MYPT-1, but at doses as high as 10 μM, it had no influence on the phosphorylation levels of AKT, MEK, and S6 kinase[1]. (0.003-10 μM) is effective in preventing human cancer cells' phosphorylation of the ROCK substrates MLC-2 and MYPT-1[1]. Effective anticancer efficacy against colorectal carcinoma (CRC) is demonstrated by RKI 1447. HCT-8 and HCT-116 cell growth is significantly suppressed by RKI 1447 (10-320 μM; 24 hours)[2]. A dose-dependent manner of inducing apoptosis is observed with RKI 1447 (20-80 μM; 24 hours)[2].
In vitro, RKI-1447 demonstrates potent inhibition of ROCK1 and ROCK2 enzymatic activity with IC50 values in the low nanomolar range. It inhibits ROCK-mediated phosphorylation of downstream substrates such as myosin light chain (MLC) and myosin phosphatase targeting subunit (MYPT1). In cell-based assays, RKI-1447 inhibits Rho/ROCK-mediated cell contraction, stress fiber formation, and cell migration. It has been shown to induce apoptosis and inhibit proliferation in various cancer cell lines. |
| ln Vivo |
The growth of breast tumors is inhibited in vivo by RKI 1447 (200 mg/kg; intraperitoneal; daily for 14 days)[1]. On CRC in vivo, RKI 1447 (100 mg/kg; ip; once every 3 days; for 14 days) exhibits anticancer efficacy. The mice are not physiologically harmed by RKI 1447[2].
In vivo, RKI-1447 has been evaluated in animal models of cancer, fibrosis, and cardiovascular diseases. In tumor xenograft models, RKI-1447 treatment inhibits tumor growth and metastasis by disrupting the actin cytoskeleton and reducing cell motility. In models of pulmonary fibrosis, RKI-1447 reduces collagen deposition and improves lung function. The compound has also shown efficacy in models of hypertension and glaucoma by reducing vascular resistance and intraocular pressure. |
| Enzyme Assay |
In vitro enzyme assays for RKI-1447 involve measuring the inhibition of ROCK1 and ROCK2 kinase activity using recombinant enzymes and synthetic peptide substrates. The kinase reaction is typically performed in the presence of ATP and a fluorescently labeled or radiolabeled substrate, and the amount of phosphorylated product is quantified. IC50 values are determined from dose-response curves. Selectivity profiling against other kinases is performed to assess the compound's specificity.
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| Cell Assay |
Cell Viability Assay[2]
Cell Types: CRC cell lines HCT-8 and HCT-116 cells Tested Concentrations: 0, 10, 20, 40, 80, 160, 320 μM Incubation Duration: 24 hrs (hours) Experimental Results: HCT-8 and HCT-116 viability was drastically diminished in a dose-dependent manner. Apoptosis Analysis[2] Cell Types: CRC cell lines HCT-8 and HCT-116 cells Tested Concentrations: 0, 20, 40, 80 μM Incubation Duration: 24 hrs (hours) Experimental Results: Treatment promoted apoptosis. Western Blot Analysis[1] Cell Types: MDA-MB-231 human breast cancer cells Tested Concentrations: 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3,10 μM Incubation Duration: Experimental Results: diminished the levels of P-MLC-2, but not total MLC-2, in a concentration-dependent manner with significant effects starting at 100 nM. Cell-based assays for RKI-1447 include measurement of ROCK-mediated phosphorylation of downstream targets such as MYPT1 and MLC by Western blot analysis. Cell migration and invasion assays (e.g., Boyden chamber, wound healing) are used to assess the functional consequences of ROCK inhibition. Cell proliferation and apoptosis assays (e.g., MTT, Annexin V staining) are performed to evaluate the compound's effects on cell viability and death. |
| Animal Protocol |
Animal/Disease Models: MMTV/neu transgenic mice [FVB/N-Tg (MMTVneu) 202 Mul/J][1]
Doses: 200 mg/kg Route of Administration: Treated ip daily for 14 days Experimental Results: Tumors from mice treated with vehicle increased in size with an average percent change in tumor volume of 68.3%. In contrast, tumors from mice treated with the RKI-1447 increased in size with an average percent change in tumor volume of only 8.8%. Thus, RKI-1447 inhibited mammary tumor growth by 87%. Animal/Disease Models: 5weeks old Male BALB/C nude mice[2] Doses: 100 mg/kg Route of Administration: intraperitoneally (ip) injected; once every 3 days; for 14 days Experimental Results: Efficiently blocked CRC tumor growth in vivo. In vivo animal studies for RKI-1447 are conducted in various disease models. In tumor xenograft models, immunocompromised mice are implanted with cancer cells and treated with RKI-1447 via oral or intraperitoneal administration. Tumor size, metastasis, and survival are monitored. In fibrosis models, mice are treated with RKI-1447 and tissue samples are analyzed for collagen content and inflammatory markers. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of RKI-1447 are not extensively documented. As a small molecule kinase inhibitor, it is expected to have moderate oral bioavailability and tissue distribution. The compound's molecular weight and lipophilicity influence its absorption, distribution, metabolism, and excretion. For research use, the compound is typically formulated in appropriate vehicles for in vivo administration and stored at -20°C.
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| Toxicity/Toxicokinetics |
Toxicological data for RKI-1447 are limited. As a ROCK inhibitor, it may have potential effects on vascular tone and platelet function due to the role of ROCK in smooth muscle contraction and platelet activation. However, specific toxicology studies are not publicly available. For research use only, not for therapeutic or human use.
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| References |
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| Additional Infomation |
RKI-1447 dihydrochloride (CAS# 1782109-09-4) is a potent and selective inhibitor of ROCK1 and ROCK2. It is used in research to study ROCK signaling in cytoskeletal dynamics, cell migration, proliferation, and apoptosis. The compound has been investigated for potential therapeutic applications in cancer, cardiovascular diseases, and fibrotic disorders. It is for research use only and not for therapeutic or human use.
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| Molecular Formula |
C16H16CL2N4O2S
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|---|---|
| Molecular Weight |
399.29
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| Exact Mass |
398.037
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| CAS # |
1782109-09-4
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| Related CAS # |
RKI-1447;1342278-01-6
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| PubChem CID |
90489001
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| Appearance |
Off-white to light yellow solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
25
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| Complexity |
392
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC(=CC(=C1)O)CNC(=O)NC2=NC(=CS2)C3=CC=NC=C3.Cl.Cl
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| InChi Key |
GBXLRAPHQIRDNS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H14N4O2S.2ClH/c21-13-3-1-2-11(8-13)9-18-15(22)20-16-19-14(10-23-16)12-4-6-17-7-5-12;;/h1-8,10,21H,9H2,(H2,18,19,20,22);2*1H
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| Chemical Name |
1-[(3-hydroxyphenyl)methyl]-3-(4-pyridin-4-yl-1,3-thiazol-2-yl)urea;dihydrochloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 50 mg/mL (125.22 mM)
H2O : < 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.26 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.26 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5044 mL | 12.5222 mL | 25.0445 mL | |
| 5 mM | 0.5009 mL | 2.5044 mL | 5.0089 mL | |
| 10 mM | 0.2504 mL | 1.2522 mL | 2.5044 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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