| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: 5.88 μM (the binding of NGF to tyrosine kinase (Trk) A and TrkA); 3.72 μM (the binding of NGF to p75 and TrkA)[1].
NGF receptor (tyrosine kinase TrkA and p75 neurotrophin receptor). ALE-0540 inhibits the binding of NGF to both TrkA and p75 receptors. TrkA is a receptor tyrosine kinase that mediates the survival, differentiation, and pain-sensitizing effects of NGF through activation of downstream signaling pathways including MAPK/ERK, PI3K/AKT, and PLCγ. Upon NGF binding, TrkA undergoes autophosphorylation and recruits signaling adaptors that propagate the signal. p75NTR is a member of the tumor necrosis factor receptor superfamily that can modulate TrkA signaling, mediate apoptosis in certain contexts, and contribute to pain signaling through interactions with other receptors. ALE-0540 antagonizes NGF binding to both receptors, thereby blocking NGF-mediated signal transduction and biological responses. |
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| ln Vitro |
ALE-0540 inhibits the binding of NGF to tyrosine kinase (Trk) A with an IC50 of 5.88 μM and to both p75 and TrkA with an IC50 of 3.72 μM. The lower IC50 for the p75/TrkA binding (3.72 μM) compared to TrkA alone (5.88 μM) suggests that ALE-0540 may have differential effects on the two receptor populations or that the presence of p75 enhances the inhibitory activity. These in vitro binding data indicate that ALE-0540 is a moderately potent NGF receptor antagonist that can disrupt the interaction between NGF and its receptors at micromolar concentrations. By blocking NGF binding, ALE-0540 prevents the downstream signaling cascades that lead to neuronal sensitization and pain transmission.
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| ln Vivo |
ALE-0540 blocks NGF-mediated signaling pathways and biological responses via TrkA receptor antagonism. In vivo, ALE-0540 has been characterized for its antiallodynic actions in rat models. In a study published in the Journal of Pharmacology and Experimental Therapeutics, ALE-0540 demonstrated antiallodynic effects in rats, suggesting that NGF receptor antagonism can alleviate pain behaviors in preclinical models. Allodynia refers to pain resulting from a stimulus that does not normally provoke pain, a common symptom of neuropathic pain conditions. The efficacy of ALE-0540 in reversing allodynia supports the concept that NGF signaling through TrkA and p75 receptors contributes to pain sensitization, and that pharmacological blockade of these receptors can provide analgesic benefits. However, detailed in vivo efficacy data including dose-response relationships and time course of action are not extensively reported in the available literature.
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| Enzyme Assay |
Receptor binding assays are performed using radiolabeled NGF (e.g., 125I-NGF) incubated with membrane preparations expressing TrkA or p75 receptors in the presence of varying concentrations of ALE-0540. The membranes are typically prepared from cells or tissues that endogenously express these receptors or from recombinant expression systems. After incubation to allow binding equilibrium, the membrane-bound radioactivity is separated from free radioligand by filtration through glass fiber filters or by centrifugation. The retained radioactivity is measured using a gamma counter, and the inhibition of specific binding is calculated. IC50 values are determined by fitting the competition binding data to a sigmoidal dose-response curve. Non-specific binding is determined in the presence of excess unlabeled NGF. This assay format allows quantitative assessment of the compound's affinity for the NGF receptors and its ability to compete with the natural ligand.
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| Cell Assay |
Cell-based assays using neuronal cell lines (e.g., PC12 cells, which express TrkA receptors and respond to NGF by differentiating into neuron-like cells) are treated with ALE-0540 at various concentrations prior to or concurrently with NGF stimulation. NGF-induced neurite outgrowth (quantified by measuring neurite length or the percentage of cells with neurites), cell survival (assessed by viability assays), or downstream signaling (e.g., phosphorylation of ERK, AKT, or PLCγ measured by Western blotting or phospho-specific ELISAs) are measured to assess functional antagonism of TrkA-mediated responses. The ability of ALE-0540 to inhibit these NGF-induced effects confirms its activity as a functional antagonist at the receptor level, distinguishing it from compounds that only bind but do not block signaling.
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| Animal Protocol |
Animal models of pain (e.g., inflammatory pain models such as carrageenan-induced hyperalgesia, or neuropathic pain models such as chronic constriction injury or spinal nerve ligation in rodents) are used to assess the in vivo analgesic efficacy of ALE-0540. ALE-0540 is administered via systemic routes (e.g., intraperitoneal, subcutaneous, or oral administration) at various dose levels. Pain behavior is assessed using standardized tests: thermal hyperalgesia (e.g., paw withdrawal latency to a thermal stimulus using the Hargreaves apparatus), mechanical allodynia (e.g., paw withdrawal threshold to von Frey filaments), and other behavioral measures. The antiallodynic actions of ALE-0540 have been characterized in rat models. The time course of effect and dose-response relationships are typically evaluated to determine efficacy and potency in vivo.
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| ADME/Pharmacokinetics |
Predicted to have moderate oral bioavailability with reasonable plasma exposure following systemic administration based on its physicochemical properties (molecular weight 301.25, moderate lipophilicity). Metabolic clearance is likely mediated by hepatic CYP450 enzymes, with potential for oxidative metabolism of the hydroxyethylamino and nitro groups. Standard pharmacokinetic profiling in rodents includes determination of Cmax, Tmax, AUC, t1/2, clearance, and volume of distribution following both intravenous and oral administration at relevant doses. Formulation in suitable vehicles (e.g., DMSO, PEG400, or cyclodextrin-based formulations) may be required to achieve adequate exposure for in vivo efficacy studies. Detailed PK data specific to ALE-0540 are not extensively reported in the available literature.
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| Toxicity/Toxicokinetics |
Expected to have a manageable safety profile at therapeutic doses based on its receptor-targeted mechanism, with potential for mechanism-related effects due to NGF receptor blockade. NGF is important for neuronal development and maintenance, but in adult animals, NGF receptor antagonism is generally well tolerated at doses that provide analgesic efficacy. However, complete blockade of NGF signaling could potentially affect sympathetic and sensory neuron function, and there is evidence that NGF plays a role in tissue repair and immune function. Preclinical toxicology studies (acute and repeat-dose) in rodents are typically required to establish the safety margin and identify potential off-target effects before advancing to clinical development. As a research compound, detailed toxicology data are not publicly available.
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| References | |
| Additional Infomation |
Structure in the first source
ALE-0540 is a research-grade NGF receptor antagonist used in neuroscience and pain research. Chemical name: 2-(2-hydroxyethylamino)-5-nitrobenzo[de]isoquinoline-1,3-dione. Molecular formula: C14H11N3O5, molecular weight: 301.25. Appearance: solid. It is typically stored as a powder at -20°C for 3 years or at 4°C for 2 years, and in solution at -80°C for 6 months or at -20°C for 1 month. ALE-0540 can be used to examine mechanisms leading to the development of agents for the treatment of pain. Synonyms: ALE-0540 is also known as 2-(2-hydroxyethylamino)-5-nitrobenzo[de]isoquinoline-1,3-dione. For research use only, not for human use. |
| Molecular Formula |
C14H11N3O5
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|---|---|
| Molecular Weight |
301.25
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| Exact Mass |
301.07
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| CAS # |
234779-34-1
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| PubChem CID |
2826530
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
0.992
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
22
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| Complexity |
491
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC2=CC(=CC3=C2C(=C1)C(=O)N(C3=O)NCCO)[N+](=O)[O-]
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| InChi Key |
SURCGQGDUADKBL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H11N3O5/c18-5-4-15-16-13(19)10-3-1-2-8-6-9(17(21)22)7-11(12(8)10)14(16)20/h1-3,6-7,15,18H,4-5H2
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| Chemical Name |
2-(2-hydroxyethylamino)-5-nitrobenzo[de]isoquinoline-1,3-dione
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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) |
DMSO :~125 mg/mL (~414.94 mM; with sonication)
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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 | 3.3195 mL | 16.5975 mL | 33.1950 mL | |
| 5 mM | 0.6639 mL | 3.3195 mL | 6.6390 mL | |
| 10 mM | 0.3320 mL | 1.6598 mL | 3.3195 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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