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Netarsudil 2HCl (formerly AR13324; AR 13324; AR-13324; Rhopressa), the dihydrochloride salt of Netarsudil, is ROCK inhibitor with antihypertensive activity and the potential for glaucoma treatment. It inhibits ROCK kinase with a Ki of 0.2-10.3 nM, also inhibits norepinephrine transport activity which can reduce the production of aqueous humor. As of 2018, Netarsudil was approved by FDA for the treatment of glaucoma and ocular hypertension.
Physicochemical Properties
| Molecular Formula | C28H27N3O3.2HCL |
| Molecular Weight | 526.45 |
| Exact Mass | 525.158 |
| Elemental Analysis | C, 63.88; H, 5.55; Cl, 13.47; N, 7.98; O, 9.12 |
| CAS # | 1253952-02-1 |
| Related CAS # | Netarsudil dimesylate;1422144-42-0; 1254032-66-0; 1253952-02-1 (HCl) |
| PubChem CID | 66599892 |
| Appearance | Typically exists as white to off-white solids at room temperature |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 36 |
| Complexity | 678 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | O(C(C1C=CC(C)=CC=1C)=O)CC1C=CC(=CC=1)[C@H](C(NC1C=CC2C=NC=CC=2C=1)=O)CN.Cl.Cl |
| InChi Key | LDKTYVXXYUJVJM-FBHGDYMESA-N |
| InChi Code | InChI=1S/C28H27N3O3.2ClH/c1-18-3-10-25(19(2)13-18)28(33)34-17-20-4-6-21(7-5-20)26(15-29)27(32)31-24-9-8-23-16-30-12-11-22(23)14-24;;/h3-14,16,26H,15,17,29H2,1-2H3,(H,31,32);2*1H/t26-;;/m1../s1 |
| Chemical Name | [4-[(2S)-3-Amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl]phenyl]methyl 2,4-dimethylbenzoate dihydrochloride |
| Synonyms | AR13324 HCl; Rhopressa; AR-13324 HCl; AR 13324; AR 13324 HCl; Netarsudil; AR-13324; Netarsudil hydrochloride; Netarsudil dihydrochloride; 1253952-02-1; AR-13324 hydrochloride; Netarsudil (hydrochloride); SE030PF6VE; AR-13324 HCL; Netarsudil (AR-13324) 2HCl; (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride; |
| 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) |
Biological Activity
| Targets |
Rho-associated protein kinas/ROCK; norepinephrine transporter/NET
Netarsudil 2HCl (AR-13324) targets Rho-associated protein kinase 1 (ROCK1) (IC50 = 0.03 nM) [1] Netarsudil 2HCl (AR-13324) targets Rho-associated protein kinase 2 (ROCK2) (IC50 = 0.02 nM) [1] Netarsudil 2HCl (AR-13324) targets norepinephrine transporter (NET) (Ki = 1.6 nM) [1] |
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| ln Vitro |
In vitro activity: Previous study showed that at the cellular level, netarsudil had been shown to be able to induce loss of actin stress fibers, cell shape changes, loss of focal adhesions, as well as changes in extracellular matrix composition of TM cells Kinase Assay: Netarsudil (formerly known as AR-13324) is ROCK inhibitor with Ki of 0.2-10.3 nM. It also inhibits norepinephrine transport activity which can reduce the production of aqueous humor. Cell Assay: Previous study showed that at the cellular level, netarsudil had been shown to be able to induce loss of actin stress fibers, cell shape changes, loss of focal adhesions, as well as changes in extracellular matrix composition of TM cells. In isolated human trabecular meshwork (TM) cells, Netarsudil 2HCl (0.1–100 nM) dose-dependently reduces ROCK-mediated phosphorylation of myosin light chain (MLC) (Western blot), with a 68% reduction at 10 nM [1] - It inhibits NET-mediated norepinephrine (NE) uptake in human embryonic kidney (HEK293) cells expressing human NET: IC50 = 3.2 nM, reducing NE uptake by ~75% at 10 nM [1] - In isolated porcine ciliary arteries precontracted with phenylephrine, Netarsudil 2HCl (0.01–10 μM) induces vasodilation (EC50 = 0.21 μM), achieving 95% relaxation at 10 μM [1] - It shows no significant cytotoxicity to human corneal epithelial cells or TM cells at concentrations up to 10 μM (cell viability >90% vs. control) [1] |
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| ln Vivo |
In normotensive monkey eyes, netarsudil hydrochloride (0.04%, 50 µL) lowers intraocular pressure (IOP)[1]. In Dutch Belted rabbits, netarsudil hydrochloride (0.04%) results in significantly decreased significanting of episcleral venous pressure (EVP)[2]. In normotensive cynomolgus monkeys: Topical administration of Netarsudil 2HCl (0.04% eye drops) twice daily for 28 days reduces intraocular pressure (IOP) by ~28% vs. vehicle. It increases aqueous humor outflow facility by ~42% and does not affect aqueous humor production [1] - In Dutch belted rabbits with elevated episcleral venous pressure (EVP): Topical Netarsudil 2HCl (0.04%, 0.12% eye drops) twice daily for 14 days dose-dependently reduces EVP. The 0.12% dose lowers EVP by ~25% vs. vehicle, with a corresponding IOP reduction of ~22% [2] - In rabbits, Netarsudil 2HCl (0.12% eye drops) increases ocular blood flow in the anterior segment (by ~30%) and optic nerve head (by ~28%) without systemic hemodynamic effects [2] |
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| Enzyme Assay | A total of 23 ROCK structures were found in the PDB. The maximum and minimum resolutions were 3.4 Å and 2.93 Å, respectively. Seven ROCK-I and two ROCK-II non-redundant structures were selected for the binding assay. Out of 46 compounds tested (20 isoquinolines, 15 aminofurazan, 6 benzodiazepine, 4 indazoles, and 1 amide), 34 presented a significantly higher docking score for ROCK-1, when compared to Y-27632 (p < 0.0001). All ROCKi classes presented a stronger mean docking score than Y-27632 (p < 0.0001). The frequency of compounds presenting highest docking score was higher in the isoquinoline, aminofurazan, and benzodiazepine classes for ROCK-I; and in isoquinolines and amides for ROCK-II (Supplementary Figure S2A). The top ten compounds that presented the highest mean docking scores for ROCK-I and II are shown in Supplementary Figure S2B. The isoquinoline class represented 70% of the drugs within the top ten highest docking scores, with three compounds presenting a docking score stronger than |
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