| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| Other Sizes |
| Targets |
ROCK/PKC; metabolite of Netarsudil
AR-13503 targets Rho-associated protein kinase (ROCK), specifically both ROCK1 and ROCK2 isoforms. ROCK is a serine/threonine kinase that is activated by the small GTPase RhoA and plays a central role in the regulation of the actin cytoskeleton. By inhibiting ROCK activity, AR-13503 prevents the phosphorylation of downstream targets including myosin light chain (MLC), LIM kinase, and cofilin, leading to reduced actomyosin contractility, altered cell migration, and changes in gene expression. This mechanism underlies its potential therapeutic effects in various diseases involving ROCK dysregulation. |
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| ln Vitro |
Human venous vascular endothelial cell (HUVEC) tube formation is inhibited by AR-13503 (16 hours) at an IC50 of 21 nM[1]. In a dose-dependent approach, AR-13503 (400 nM; 5 days) decreases the sprouting area of venous vasculature [1]. In a dose-dependent manner, AR-13503 (400 nM; 2 weeks) improves the primary RPE barrier function in pigs [1].
In vitro, AR-13503 demonstrates potent ROCK inhibitory activity with IC₅₀ values in the low nanomolar range for both ROCK1 and ROCK2. It inhibits ROCK-mediated phosphorylation of myosin light chain and other downstream substrates in various cell types, including smooth muscle cells, endothelial cells, and cancer cells. It shows anti-proliferative effects in cancer cell lines and inhibits cell migration and invasion. It reduces endothelial permeability and has anti-angiogenic effects. In vascular smooth muscle cells, it inhibits contraction and promotes vasodilation. It also shows anti-fibrotic effects by inhibiting fibroblast activation and collagen production. |
| ln Vivo |
In combination with Aflibercept, AR-13503 (1.25 mg/kg; intraperitoneal injection; once daily for 5 days) effectively inhibits the development of aberrant neovascularization (NV) [1].
In vivo, AR-13503 has shown efficacy in various animal models. In models of glaucoma, it lowers intraocular pressure by improving aqueous humor outflow. In cardiovascular models, it reduces blood pressure and improves vascular function. In cancer models, it inhibits tumor growth and metastasis by reducing tumor cell migration and invasion, and by affecting the tumor microenvironment. In fibrotic disease models, it reduces tissue fibrosis by inhibiting fibroblast activation and extracellular matrix deposition. Its effects are consistent with ROCK inhibition and demonstrate the broad therapeutic potential of ROCK targeting. |
| Enzyme Assay |
In vitro ROCK kinase assays for AR-13503 use purified recombinant ROCK1 or ROCK2 enzymes. The kinase assay is typically performed in 96-well plates using a kinase reaction buffer (50 mM HEPES, pH 7.4, 10 mM MgCl₂, 1 mM DTT, 0.01% Triton X-100) with ATP and a peptide substrate (e.g., myosin light chain peptide or S6 kinase substrate). Various concentrations of AR-13503 (0.01 nM to 10 μM) are incubated with the enzyme and substrate, and the reaction is initiated by adding ATP. After incubation at 30°C for 30-60 minutes, the reaction is stopped, and phosphorylation is detected using a fluorescence-based or luminescence-based assay (e.g., ADP-Glo, TR-FRET). IC₅₀ values are calculated from dose-response curves. Selectivity against other kinases is assessed in panel screening.
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| Cell Assay |
In vitro cell-based assays for AR-13503 use various cell types including vascular smooth muscle cells, endothelial cells, cancer cell lines, and fibroblasts. Cells are cultured in appropriate media (DMEM or RPMI-1640 with 10% FBS, 37°C, 5% CO₂) and treated with AR-13503 at various concentrations (0.001-100 μM) for 1-72 hours. ROCK activity is assessed by measuring phosphorylation of myosin light chain (MLC) and other ROCK substrates by Western blot. Cell proliferation is measured by MTT, SRB, or CellTiter-Glo assays. Cell migration is assessed using scratch wound healing or transwell migration assays. Cell invasion is measured using Matrigel-coated transwell inserts. Apoptosis is evaluated by caspase-3/7 activation and Annexin V/PI staining. Cytotoxicity is determined by LDH release assays.
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| Animal Protocol |
Animal/Disease Models: Oxygen-induced retinopathy (OIR) mouse model (C57BL/6; 7 days old) [1]
Doses: 1.25 mg/kg Route of Administration: intraperitoneal (ip) injection; one time/day for 5 days Experimental Results: Combination treatment group The reduction in NV (approximately 75%) was greater than that in the aflibercept alone group (approximately 55%). In the oxygen-induced retinopathy (OIR) animal model, 7-day old neonatal C57BL/6 mice were housed with foster mothers at 75% oxygen from postnatal day (P)7 to P12. Upon return to normoxia on P12, mice were treated by once daily intraperitoneal injections with vehicle control, aflibercept, or AR-13503 in combination with aflibercept. Tissues were harvested following 5 days of treatment and stained for isolectin-IB4 for visualization and analysis of NV. Retinas were also harvested for pharmacokinetic analysis.[2] In vivo animal studies for AR-13503 are conducted in various disease models. For glaucoma studies, rabbits or mice are treated with AR-13503 topically (eye drops) or by intravitreal injection, and intraocular pressure is measured using a tonometer. For cardiovascular studies, spontaneously hypertensive rats or other models are treated with AR-13503 orally or intraperitoneally (1-30 mg/kg), and blood pressure is monitored by tail-cuff or telemetry. For cancer studies, tumor xenograft models are used with AR-13503 administration (oral or IP, daily or on a schedule) and tumor volume measurement. For fibrosis studies, models of pulmonary, hepatic, or renal fibrosis are used with AR-13503 treatment and histological assessment of fibrosis. |
| ADME/Pharmacokinetics |
The pharmacokinetic properties of AR-13503 have been characterized in preclinical studies. As a small molecule kinase inhibitor, it shows good oral bioavailability and tissue distribution. Following administration, it is absorbed and distributed to target tissues including the eye, vasculature, and tumors. The compound is metabolized in the liver, likely through cytochrome P450-mediated oxidation and conjugation. Its elimination half-life supports once or twice daily dosing in animal studies. The compound shows good brain penetration, which may be relevant for central nervous system applications. Specific PK parameters vary by species and formulation.
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| Toxicity/Toxicokinetics |
AR-13503 has been evaluated for toxicity in preclinical studies. As a ROCK inhibitor, it is generally well-tolerated at therapeutic doses. Common adverse effects may include hypotension, headache, and gastrointestinal disturbances due to the physiological role of ROCK in vascular tone and smooth muscle function. At high doses, more significant effects on blood pressure and organ perfusion may occur. The compound is not associated with significant hepatotoxicity or nephrotoxicity at effective doses. Comprehensive toxicological studies are needed for therapeutic development. This compound is for research use only and is not approved for human use.
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| References |
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| Additional Infomation |
Diabetic retinopathy (DR) and its complications can lead to irreversible vision loss. Due to the limitations of anti-vascular endothelial growth factor (VEGF) and steroid therapy, there is an urgent need for new therapies with novel mechanisms of action. Rho kinase (ROCK) is closely associated with retinal neovascularization (NV) and vascular leakage in neovascular age-related macular degeneration (nAMD) and proliferative diabetic retinopathy (PDR). AR-13503 is a potent ROCK and protein kinase C (PKC) inhibitor. We have demonstrated in vitro and in vitro experiments that AR-13503 can inhibit angiogenesis and enhance the function of the retinal pigment epithelium barrier. This study investigated the effect of AR-13503 on abnormal neovascularization (NV) formation in an animal model of proliferative diabetic retinopathy (PDR). [2] Results: AR-13503 was detectable in the retina of mice after systemic administration of 1.25 mg/kg/day. Compared with the vector control group, mice treated with aflibercept alone had approximately 55% less NV. AR-13503 combined with aflibercept resulted in a greater reduction in NV (approximately 75%), which was superior to aflibercept alone. There was no significant difference in avascular area among the groups. [2]
AR-13503 (CAS# 1254032-16-0) is a small molecule research compound that acts as a Rho kinase (ROCK) inhibitor, targeting both ROCK1 and ROCK2 isoforms. It has been investigated for potential applications in cardiovascular disease, glaucoma, cancer, and fibrotic diseases. The compound is for research use only and is not approved for clinical use. It is available from chemical suppliers for research purposes. Standard safety precautions should be followed when handling this compound. This product is for research use only, not for human therapeutic use. |
| Molecular Formula |
C19H19N3O2
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|---|---|
| Exact Mass |
321.147
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| Elemental Analysis |
C, 71.01; H, 5.96; N, 13.08; O, 9.96
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| CAS # |
1254032-16-0
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| Related CAS # |
1422144-42-0 (mesylate);1254032-66-0;1253952-02-1 (HCl);1254032-16-0;
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| PubChem CID |
134128281
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| Appearance |
White to off-white solid powder
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| LogP |
1.6
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
24
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| Complexity |
409
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O=C([C@H](CN)C1C=CC(CO)=CC=1)NC1C=CC2C=NC=CC=2C=1
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| InChi Key |
LTXBFJFJUIJOQE-GOSISDBHSA-N
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| InChi Code |
InChI=1S/C19H19N3O2/c20-10-18(14-3-1-13(12-23)2-4-14)19(24)22-17-6-5-16-11-21-8-7-15(16)9-17/h1-9,11,18,23H,10,12,20H2,(H,22,24)/t18-/m1/s1
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| Chemical Name |
(2S)-3-amino-2-[4-(hydroxymethyl)phenyl]-N-isoquinolin-6-ylpropanamide
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| Synonyms |
AR-11324 metabolite; AR13503; AR11324 metabolite; AR-13324 M1 metabolite; AR-13503; 2309668-15-1; 4MVW7F3BCT; Netarsudil metabolite AR-13503; UNII-4MVW7F3BCT; 1254032-16-0; CHEMBL4753043; Netarsudil metabolite; AR 11324 metabolite; AR-13503 AR 13503
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: ~40 mg/mL (~124.5 mM; ultrasonic and warming and adjust pH to 2 with 1 M HCl and heat to 60°C)
1 M HCl: ~25 mg/mL (~77.8 mM; with ultrasonication) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 4 mg/mL (12.45 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 DMSO stock solution (40.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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: ≥ 4 mg/mL (12.45 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 DMSO stock solution (40.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. 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. Solubility in Formulation 3: ≥ 4 mg/mL (12.45 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 DMSO stock solution (40.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
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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