| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Targets |
NRL-1049 dihydrochloride specifically and selectively targets the rho‑associated protein kinase 2 (ROCK2) enzyme. The compound is 44‑fold more selective for ROCK2 than ROCK1. It inhibits the phosphorylation activity of ROCK2, which plays a crucial role in regulating the actin cytoskeleton, cell contraction, and endothelial barrier function. This inhibition is the basis for its protective effects on the blood‑brain barrier.
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| ln Vitro |
In vitro, NRL-1049 dihydrochloride is a selective ROCK2 inhibitor with an IC50 of 0.59 uM, while showing significantly less activity against ROCK1 (IC50 = 26 uM). The compound effectively reduces lysophosphatidic acid‑induced ROCK activation in endothelial cells. It is not intended to be used directly in assays as an active drug but is evaluated in vitro for its target selectivity and potency against the recombinant ROCK2 enzyme. It shows low nanomolar to low micromolar activity against its target.
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| ln Vivo |
In vivo, NRL-1049 dihydrochloride demonstrates significant therapeutic effects in multiple mouse models of neurological injury. In a cavernous angioma model, it reduces lesion volume and hemorrhagic transformation. It also preserves blood‑brain barrier integrity and suppresses epileptic seizures after brain injury. Furthermore, in a mouse model of ischemic stroke, the compound inhibits hemorrhagic transformation, supporting its development as a drug candidate for cerebral cavernous malformations and other central nervous system disorders.
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| Enzyme Assay |
The primary in vitro assay for this compound is a kinase inhibition assay. Recombinant human ROCK2 (or ROCK1) enzyme is incubated with NRL-1049 dihydrochloride in a kinase buffer with a substrate (e.g., a myosin light chain peptide) and ATP. After incubation, the reaction is stopped, and phosphorylation levels are measured. The concentration required to inhibit 50% of the enzymatic activity (IC50) is calculated, with values of 0.59 uM for ROCK2 and 26 uM for ROCK1. For selectivity profiling, a panel of over 100 kinases is tested.
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| Cell Assay |
For in vitro cell‑based assays, human brain endothelial cells are cultured in standard endothelial cell medium. Cells are pre‑treated with NRL-1049 dihydrochloride at various concentrations (e.g., 0.1-10 uM) for 30-60 minutes, then stimulated with lysophosphatidic acid (LPA) to induce ROCK activation. ROCK activity is measured by assessing the phosphorylation of its downstream targets (e.g., myosin light chain, MYPT1) via Western blotting. Endothelial barrier function is assessed by measuring trans‑endothelial electrical resistance (TEER) or by evaluating the permeability of the cell monolayer to fluorescently labeled molecules. Cell viability is monitored by MTT assay.
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| Animal Protocol |
In animal efficacy studies, mouse models of central nervous system injury are used. For a cortical cryoinjury model, mice (e.g., C57BL/6) are subjected to a controlled brain injury. NRL-1049 dihydrochloride is administered orally at doses of 10-30 mg/kg. The compound's ability to preserve the BBB is assessed by measuring the extravasation of Evans blue dye into the brain parenchyma. In the cavernous angioma model, lesion volume and hemorrhagic transformation are measured. For the ischemic stroke model (middle cerebral artery occlusion, MCAO), the infarct volume and extent of hemorrhagic transformation are evaluated. Seizure activity is monitored by electroencephalography (EEG). Histopathological analysis of brain tissue and measurement of tight junction protein expression are also performed.
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| ADME/Pharmacokinetics |
Following oral administration, NRL-1049 dihydrochloride exhibits favorable absorption in rodent models, achieving a Tmax of approximately 0.5-2 hours. The compound shows good oral bioavailability (F%). The terminal elimination half‑life (t1/2) is approximately 2-6 hours. It is metabolized primarily by liver cytochrome P450 enzymes and excreted in feces. It can be detected in the brain following systemic administration, consistent with its mechanism of action.
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| Toxicity/Toxicokinetics |
In preclinical studies, NRL-1049 dihydrochloride is well tolerated at therapeutic doses (10-30 mg/kg). A high selectivity for ROCK2 over ROCK1 predicts a lower risk of hypotension and other ROCK1‑related side effects observed with non‑selective ROCK inhibitors. No significant body weight loss or severe clinical signs of toxicity are reported at effective doses in mice. The compound has a favorable profile for clinical development and is not mutagenic in standard Ames tests.
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| References | |
| Additional Infomation |
NRL-1049 dihydrochloride is a research‑stage ROCK2‑selective inhibitor originally developed for cerebral cavernous malformations (CCM). ROCK2 is hyperactivated in patients with CCM, and this compound has shown efficacy in preclinical models. A New Drug (IND) application has been filed by Neurelis for this compound as a potential first therapy for cerebral cavernous malformations. It is not yet approved for clinical use. NRL-1049 is strictly for research purposes.
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| Molecular Formula |
C16H23CL2N3O2S
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| Molecular Weight |
392.34
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| CAS # |
1973494-17-5
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| Related CAS # |
NRL-1049; (Rac)-NRL-1049-d5 dihydrochloride; (Rac)-NRL-1049 dihydrochloride
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| Appearance |
White to off-white solid powder
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| Synonyms |
BA-1049
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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, avoid exposure to moisture. |
| 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 | 2.5488 mL | 12.7440 mL | 25.4881 mL | |
| 5 mM | 0.5098 mL | 2.5488 mL | 5.0976 mL | |
| 10 mM | 0.2549 mL | 1.2744 mL | 2.5488 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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