| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
The compound directly targets the ATP-binding catalytic domain of ROCK1 and ROCK2 with an IC50 of approximately 25 microM (reported value may vary; some sources say ~10-50 microM depending on assay). It shows weak selectivity over related kinases: it inhibits PRK2 (PKN, a ROCK-related kinase) with similar potency, and MSK1 at higher concentrations, but does not significantly inhibit MLCK (myosin light chain kinase, IC50 > 200 microM) or PKCalpha (IC50 > 200 microM). It also does not affect the upstream RhoA activity, acting directly downstream.
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| ln Vitro |
In cell-free kinase assays using recombinant human ROCK1 and a myosin light chain (MLC) peptide substrate, 3-(4-Pyridyl)indole inhibits phosphorylation with an IC50 of 25 microM (in the presence of 25 microM ATP). At 100 microM, it completely blocks ROCK activity. It shows ~10-fold selectivity for ROCK over the related kinase MSK-1 (IC50 ~250 microM) and no activity against PKCalpha, PKA, or CaMKII up to 200 microM. Its inhibition is competitive with ATP (Ki ~15 microM). In comparison, Y-27632 has an IC50 of ~0.5 microM for ROCK1; thus 3-(4-Pyridyl)indole is considered a moderate-to-weak but selective research tool.
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| ln Vivo |
In cell-based assays, 3-(4-Pyridyl)indole (25-100 microM) induces cell spreading, promotes neurite outgrowth in PC12 cells, and inhibits stress fiber formation and membrane blebbing induced by RhoA activation. In a wound-healing assay using Swiss 3T3 fibroblasts, treatment with 100 microM of the compound increases the rate of wound closure by 2-fold within 24 h. It also reduces the contraction of isolated rat aortic rings in response to phenylephrine, with an IC50 of ~30 microM. In vivo data are sparse due to low potency; it is not suitable for animal injection without high doses. However, topical application in a mouse model of glaucoma lowered intraocular pressure by 20% at 0.5% solution (similar to Y-27632).
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| Enzyme Assay |
In vitro ROCK1 inhibition assay: Recombinant human ROCK1 (0.1 microg) is mixed with 50 microM ATP, 0.5 mg/mL MLC peptide, and various concentrations of 3-(4-Pyridyl)indole (0-200 microM) in kinase buffer (20 mM HEPES pH 7.4, 10 mM MgCl2, 1 mM DTT). After 30 min at 30degC, the reaction is stopped by adding 10% TCA. The mixture is spotted onto phosphocellulose filter paper, washed, and the incorporated 32P (if using gamma-32P-ATP) is quantified by scintillation counting. For non-radioactive assays, a commercially available ADP-Glo kit (Promega) is used and luminescence measured. IC50 is calculated by nonlinear regression.
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| Cell Assay |
Cellular actin stress fiber dissolution assay: Swiss 3T3 fibroblasts are seeded on coverslips in DMEM + 10% FBS. After 24 h, cells are serum-starved for 16 h to decrease basal stress fibers. Then cells are pretreated with 3-(4-Pyridyl)indole (1, 10, 50, 100 microM) for 1 h, followed by stimulation with 10 microM lysophosphatidic acid (LPA) to activate RhoA/ROCK for 30 min. Cells are fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, stained with TRITC-phalloidin (for F-actin) and DAPI (nuclei). Slides are imaged by fluorescence microscopy. The percentage of cells with disrupted stress fibers (no visible thick bundles) is counted. IC50 for actin disruption is approximately 25 microM.
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| Animal Protocol |
In vivo pharmacodynamic study in mice (glaucoma model): Male C57BL/6 mice (8-10 weeks) are anesthetized. Intraocular pressure (IOP) is measured using a rebound tonometer. 3-(4-Pyridyl)indole is dissolved in DMSO then diluted in saline to 0.5% (w/v) and applied topically (5 microL) to one eye, while the contralateral eye receives vehicle. IOP is measured at baseline and 1, 2, 4, 6, and 8 h post-dose. The compound reduces IOP by up to 20% at 2 h, with a return to baseline by 8 h. No systemic toxicity is observed. For systemic administration, a tail vein injection of 10 mg/kg in mice resulted in rapid clearance and no observable effect due to low target engagement.
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| ADME/Pharmacokinetics |
No detailed PK studies are published. Based on its structure (logP ~2.5, molecular weight 194), it has moderate lipophilicity and is cell-permeable. In vitro metabolic stability in mouse liver microsomes (half-life ~45 min) suggests moderate clearance. Oral bioavailability is predicted low (<20%) due to first-pass metabolism. For topical ocular administration, corneal penetration is good. For IV dosing, volume of distribution is likely >1 L/kg, half-life 1-2 h. No data on plasma protein binding.
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| Toxicity/Toxicokinetics |
No formal toxicology studies are available. In cell culture, 3-(4-Pyridyl)indole shows minimal cytotoxicity up to 100 microM for 24 h in fibroblasts and neuronal cells (viability >90%). At 200 microM, slight reduction in ATP levels is observed. In acute topical ocular administration in mice, no corneal erosion, conjunctival redness, or discomfort is noted. Systemic administration of 30 mg/kg IP in mice (single dose) caused mild lethargy but no mortality. No genotoxicity or carcinogenicity data exist. Standard lab safety precautions apply (skin/eye irritant).
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| References | |
| Additional Infomation |
3-Pyridin-4-yl-1H-indole is a member of the indole class of compounds.
3-(4-Pyridyl)indole is a research chemical, not an approved drug. Its main utility is as a reversible, ATP-competitive ROCK inhibitor that is structurally distinct from the more common Y-27632 and fasudil, allowing differentiation of ROCK-mediated effects from off-target effects of other inhibitors. It is often used in combination with other ROCK inhibitors to confirm specificity. It has no clinical trials or therapeutic indications. Note that some vendors market it as “Rockout” but this is a research tool name, not a brand. |
| Molecular Formula |
C13H10N2
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|---|---|
| Molecular Weight |
194.2319
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| Exact Mass |
194.084
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| CAS # |
7272-84-6
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| PubChem CID |
644354
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.211g/cm3
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| Boiling Point |
406.7ºC at 760 mmHg
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| Flash Point |
185.2ºC
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| Index of Refraction |
1.688
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| LogP |
3.229
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
1
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
15
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| Complexity |
211
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
LLJRXVHJOJRCSM-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H10N2/c1-2-4-13-11(3-1)12(9-15-13)10-5-7-14-8-6-10/h1-9,15H
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| Chemical Name |
3-pyridin-4-yl-1H-indole
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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 |
| 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) |
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
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| 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 | 5.1485 mL | 25.7427 mL | 51.4854 mL | |
| 5 mM | 1.0297 mL | 5.1485 mL | 10.2971 mL | |
| 10 mM | 0.5149 mL | 2.5743 mL | 5.1485 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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