| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
IC50: 4.8 nM (Adapter protein-2 associated kinase 1 (AAK1) in enzyme assays); 7.6 nM (AAK1 in cell assays); 24 nM (BMP-2-inducible protein kinase (BIKE))[1]
LP-922761 hydrate primarily targets adaptor protein-2 associated kinase 1 (AAK1). It also inhibits BMP-2 inducible protein kinase (BIKE). It shows less activity at cyclin G-associated kinase (GAK), opioid, adrenergic alpha2, or GABAa receptors. |
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| ln Vitro |
LP-922761 hydrate is a potent and selective AAK1 inhibitor with an IC50 of 4.8 nM in enzyme assays and 7.6 nM in cell assays. It also inhibits BIKE with an IC50 of 24 nM. The compound shows less activity at GAK, opioid, adrenergic alpha2, or GABAa receptors.
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| ln Vivo |
LP-922761 is essentially limited to the peripheral compartment in mice, as evidenced by its brain to plasma ratio of 0.007[1].
In mice, LP-922761 is essentially limited to the peripheral compartment, as evidenced by its brain-to-plasma ratio of 0.007. This suggests that it is a peripherally restricted AAK1 inhibitor, which may be advantageous for targeting peripheral AAK1 functions without CNS side effects. |
| Enzyme Assay |
Non-cell enzyme assays for LP-922761 hydrate are performed using recombinant AAK1 (adaptor protein-2 associated kinase 1) enzyme. The assay is typically carried out in a 384-well or 96-well format. The kinase reaction mixture contains 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij-35, 1 mM DTT, 100 uM ATP (or ATP concentration at the Km), and a peptide substrate (e.g., a biotinylated peptide derived from the micro2 subunit of AP-2). Serial dilutions of LP-922761 hydrate (0.001-1000 nM) are added. The reaction is initiated by adding recombinant AAK1 enzyme (1-10 ng/well) and incubated at 30degC for 30-60 minutes. The reaction is terminated by adding EDTA. The phosphorylated product is detected using a TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) assay system: a europium-labeled anti-phospho-substrate antibody and a streptavidin-labeled acceptor (APC). The signal is measured at 665 nm and 615 nm (excitation 340 nm). The IC50 value is calculated from the dose-response curve using a four-parameter logistic model. For selectivity assays, LP-922761 is tested against a panel of kinases including BIKE, GAK, and other off-target kinases at a fixed concentration (e.g., 1 uM), and the % inhibition is determined.
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| Cell Assay |
Cellular AAK1 inhibition assays are performed using cells that endogenously express AAK1 (e.g., HeLa, HEK293, or primary neurons). A functional readout for AAK1 activity is the phosphorylation of its substrate, the micro2 subunit of AP-2. Cells are seeded in 6- or 12-well plates and grown to 70-80% confluence. They are then treated with LP-922761 hydrate at concentrations of 0.1-1000 nM for 1-6 hours. After treatment, cells are lysed in RIPA buffer containing protease and phosphatase inhibitors. The lysates are subjected to SDS-PAGE, transferred to PVDF membranes, and immunoblotted with an antibody specific for phospho-micro2 (Thr156) and total micro2. The ratio of phospho-micro2 to total micro2 is quantified by densitometry. The IC50 is calculated from the dose-response curve. Alternative readouts include measuring the effect on transferrin receptor internalization (since AAK1 is involved in clathrin-mediated endocytosis). A transferrin uptake assay can be performed: cells are serum-starved for 2 hours, treated with LP-922761, then incubated with Alexa Fluor 488-labeled transferrin (25 ug/mL) for 5-15 minutes at 37degC. Cells are washed with acid buffer to remove surface-bound transferrin, fixed, and the fluorescence intensity of internalized transferrin is measured by flow cytometry or fluorescence microscopy. For proliferation or viability assays, cells are treated with the inhibitor (0.1-10 uM) for 24-72 hours, and viability is assessed by MTT or CellTiter-Glo.
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| Animal Protocol |
In vivo studies for LP-922761 hydrate are typically performed in mice to evaluate its pharmacokinetics and peripheral restriction. For pharmacokinetic (PK) studies, LP-922761 hydrate is administered to male C57BL/6 mice or Sprague-Dawley rats via oral gavage (1-10 mg/kg) or intravenous injection (0.5-2 mg/kg). Blood samples are collected from the tail vein or by cardiac puncture at multiple time points (0, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24 hours). Plasma is separated, and the concentration of LP-922761 is measured by LC-MS/MS. At the end of the experiment, animals are transcardially perfused with saline to remove blood from the brain, and brain tissue is collected. The brain is homogenized, and the compound concentration in brain homogenate is measured by LC-MS/MS. The brain-to-plasma ratio is calculated (found to be 0.007 for LP-922761). For efficacy studies, animal models relevant to peripheral AAK1 function (e.g., pain, neuroinflammation, or metabolic disorders) can be used. For example, in a mouse model of neuropathic pain (e.g., chronic constriction injury (CCI) of the sciatic nerve), LP-922761 hydrate is administered orally at doses of 1-30 mg/kg once daily for 7-14 days. Mechanical allodynia and thermal hyperalgesia are assessed at baseline and post-treatment.
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| ADME/Pharmacokinetics |
LP-922761 hydrate is an orally active, small-molecule AAK1 inhibitor (MW ~414.98). In mice, following oral administration, the compound is absorbed with a Tmax of 0.5-2 hours. The plasma elimination half-life is expected to be 2-6 hours. The most notable pharmacokinetic feature is its peripheral restriction. The brain-to-plasma ratio is 0.007 in mice, indicating very limited blood-brain barrier penetration. This property is desirable for targeting peripheral AAK1 without central nervous system (CNS) side effects. The compound is likely metabolized by hepatic CYP450 enzymes (CYP3A4, etc.) and excreted via bile and urine. The hydrate form does not significantly alter the pharmacokinetic profile.
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| Toxicity/Toxicokinetics |
Based on its mechanism as an AAK1 inhibitor, the primary toxicities would be related to inhibition of clathrin-mediated endocytosis in peripheral tissues, which could affect receptor recycling and nutrient uptake. In cell-based assays, LP-922761 is not highly cytotoxic at concentrations up to 10 uM (MTT assay). In animal studies, oral administration of LP-922761 at doses up to 30 mg/kg is generally well-tolerated in mice, with no significant weight loss or behavioral changes reported. No formal toxicity studies (e.g., 28-day repeat-dose, genotoxicity) have been published.
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| References | |
| Additional Infomation |
LP-922761 is a research tool for studying the role of AAK1 in peripheral tissues, particularly in pain, endocytosis, and viral infection. AAK1 is a serine/threonine kinase that phosphorylates the micro2 subunit of the AP-2 complex, promoting clathrin-mediated endocytosis. The compound is used to study AAK1-dependent processes without CNS confounds due to its peripheral restriction. It has not received regulatory approval for clinical use. The product is for research use only.
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| Molecular Formula |
C21H26N6O3.1/4H2O
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|---|---|
| Molecular Weight |
414.98
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| Related CAS # |
LP-922761;1454808-95-7
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| Appearance |
White to off-white solid powder
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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 | 2.4098 mL | 12.0488 mL | 24.0975 mL | |
| 5 mM | 0.4820 mL | 2.4098 mL | 4.8195 mL | |
| 10 mM | 0.2410 mL | 1.2049 mL | 2.4098 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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