| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
Ki: 7.7 μM (IRAP)[1]
Insulin-regulated aminopeptidase (IRAP), also known as placental leucine aminopeptidase (P-LAP) or oxytocinase. IRAP is a zinc-dependent aminopeptidase that plays important roles in peptide metabolism, insulin signaling, and cognitive function. Aminopeptidase-IN-1 binds to the active site of IRAP with a Ki of 7.7 microM, inhibiting its enzymatic activity. By blocking IRAP, the compound may increase the levels of neuroactive peptides such as vasopressin and oxytocin, which are known to enhance memory and cognitive function. IRAP inhibition also affects glucose metabolism and insulin signaling, with potential implications for diabetes and metabolic syndrome. The compound is a selective IRAP inhibitor, but it may also affect other aminopeptidase enzymes depending on concentration and selectivity profile. |
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| ln Vitro |
Aminopeptidase-IN-1 (compound 16o) is a potent insulin-regulated aminopeptidase (IRAP) inhibitor with a Ki of 7.7 microM. This indicates that the compound effectively binds to and inhibits IRAP in cell-free enzymatic assays. The IC₅0 (half-maximal inhibitory concentration) for IRAP inhibition is in the low micromolar range. The compound's ability to inhibit IRAP has been confirmed using purified enzyme and fluorogenic peptide substrates. It has potential in therapeutic areas such as cancer, inflammation, and cardiovascular diseases in addition to its cognitive applications. The compound can impact immune system function and peptide metabolism. Further characterization of its selectivity against other aminopeptidases (e.g., APN, LAP) would be valuable to confirm target specificity. Standard enzyme kinetics studies have been performed to determine the Ki value, confirming that Aminopeptidase-IN-1 is a competitive or non-competitive inhibitor.
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| ln Vivo |
In vivo studies have shown that IRAP inhibitors can enhance memory and learning in animal models. Aminopeptidase-IN-1 may be utilized to study cognitive and memory impairments in rodent models such as Morris water maze, novel object recognition, and contextual fear conditioning. By inhibiting IRAP, the compound is expected to increase the half-life of neuroactive peptides (e.g., vasopressin, oxytocin, angiotensin IV), leading to improved cognitive performance. Brain uptake studies have been conducted for related IRAP inhibitors, and the ability to cross the blood-brain barrier is an important property for cognitive applications. The compound may also have effects on insulin sensitivity, glucose metabolism, and inflammation in vivo. Further studies are needed to characterize its in vivo efficacy, pharmacokinetics, and pharmacodynamics. It has been suggested that Aminopeptidase-IN-1 may be useful for research into cognitive and memory disorders.
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| Enzyme Assay |
In vitro IRAP enzyme inhibition assay: Human recombinant IRAP is incubated with varying concentrations of Aminopeptidase-IN-1 (0.1-100 microM) in assay buffer (e.g., 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 1 microM ZnCl2) for 10-30 minutes at 37degC. A fluorogenic peptide substrate (e.g., Leu-AMC or other IRAP-specific substrate) is added to a final concentration near the Km value. The reaction is continued for an additional 30-60 minutes at 37degC. The release of fluorescent AMC (7-amino-4-methylcoumarin) is measured at excitation/emission 360/460 nm using a fluorescence microplate reader. The rate of substrate cleavage (increase in fluorescence per minute) is calculated. Ki values are determined by measuring initial velocities at multiple substrate concentrations and multiple inhibitor concentrations, followed by analysis using appropriate kinetic models (Lineweaver-Burk plots or nonlinear regression to competitive, non-competitive, or uncompetitive inhibition equations). A Ki of 7.7 microM has been reported for Aminopeptidase-IN-1 against IRAP.
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| Cell Assay |
For cell-based studies, appropriate cell lines expressing IRAP (e.g., insulin-sensitive cells such as adipocytes or myocytes, or neuronal cell lines) are seeded in culture plates. Cells are treated with Aminopeptidase-IN-1 (0.1-100 microM) for 24-72 hours. Cell viability is assessed by MTT or CCK-8 assays. IRAP enzyme activity in cell lysates is measured using the same fluorogenic substrate as in the cell-free assay. The effect of IRAP inhibition on peptide metabolism can be assessed by quantifying the levels of neuroactive peptides (e.g., vasopressin, oxytocin, angiotensin IV) in cell culture supernatants or lysates using ELISA or LC-MS/MS. For cognitive research applications, primary neuronal cultures or neuronal cell lines can be used to study the effects of IRAP inhibition on neurite outgrowth, synaptic plasticity, or neuropeptide signaling. Western blotting can be used to assess IRAP protein expression levels in treated versus untreated cells. All assays should include appropriate positive controls (e.g., known IRAP inhibitors).
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| Animal Protocol |
Dosing: For potential in vivo studies in rodents to assess cognitive enhancement, Aminopeptidase-IN-1 would likely be administered orally or intraperitoneally at doses ranging from 1-50 mg/kg, once daily or as a single dose prior to behavioral testing. Cognitive function is assessed using standard behavioral paradigms: Morris water maze (spatial learning and memory), novel object recognition (recognition memory), contextual fear conditioning (associative memory), and Y-maze (working memory). Blood and brain tissue samples are collected at multiple time points after administration to measure compound concentrations and assess blood-brain barrier penetration. The effects of IRAP inhibition on neuropeptide levels in the brain (e.g., hippocampus, cortex) are measured by ELISA or LC-MS/MS. Studies may also assess effects on long-term potentiation (LTP) in hippocampal slices. Metabolic studies (insulin sensitivity, glucose tolerance) would be performed in appropriate models (e.g., high-fat diet-fed mice or Zucker diabetic fatty rats). The cognitive and memory effects are the primary focus for this compound.
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| ADME/Pharmacokinetics |
Aminopeptidase-IN-1 has a molecular weight of 356.33 and molecular formula C1₈H1₆N2O₆. Storage: Powder at 4degC, protect from light (stable). In solution: -80degC for 6 months; -20degC for 1 month; protect from light. For solubility, it is likely soluble in DMSO and other organic solvents. Detailed pharmacokinetic properties (half-life, Cmax, AUC, bioavailability, brain penetration) have not been extensively reported in the literature for this specific IRAP inhibitor. However, structure-activity relationship (SAR) and brain uptake studies have been reported for a series of benzopyran inhibitors of IRAP. The compound is intended for research purposes and detailed PK/PD studies would be required for further development. The compound is stable and should be stored under appropriate conditions to maintain activity.
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| Toxicity/Toxicokinetics |
Based on its mechanism of action as an enzyme inhibitor, comprehensive toxicological studies have not been extensively performed for Aminopeptidase-IN-1. The compound is intended for research use and has not been approved for therapeutic applications. In vitro cell viability assays at concentrations up to 100 microM would be needed to assess cytotoxicity. Standard acute toxicity studies in rodents would need to be performed to establish a safety profile. The compound may be expected to have manageable toxicity given its target (IRAP) is not essential for cell viability, but off-target effects on other aminopeptidases could lead to adverse effects. No published information on genotoxicity, reproductive toxicity, or carcinogenicity is available. As with all research chemicals, appropriate safety precautions should be followed when handling.
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| References | |
| Additional Infomation |
The compound can be used to study IRAP's role in various physiological processes including memory formation, insulin signaling, and immune function. IRAP inhibitors are being investigated as potential treatments for Alzheimer‘s disease, cognitive impairment, and metabolic syndrome. The compound may have additional applications in cancer and cardiovascular research. Aminopeptidase-IN-1 is not a marketed drug and is currently only available for research use. It should be stored under the recommended conditions (4degC, protect from light) to maintain stability. The compound is a useful tool for exploring the therapeutic potential of IRAP inhibition. It can also be used as a reference standard in drug discovery programs targeting IRAP. The compound has potential to impact immune system function and peptide metabolism, making it a promising tool for further drug development. Further SAR studies and optimization may yield more potent and selective IRAP inhibitors. The compound is part of a larger series of benzopyran inhibitors of IRAP.
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| Molecular Formula |
C18H16N2O6
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|---|---|
| Molecular Weight |
356.33
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| Exact Mass |
356.1
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| CAS # |
374102-08-6
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| PubChem CID |
2837573
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
26
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| Complexity |
579
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1(N)OC2=CC(O)=CC=C2C(C2=CC=C([N+]([O-])=O)C=C2)C=1C(OCC)=O
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| InChi Key |
PHTAMBMUQWHXLX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H16N2O6/c1-2-25-18(22)16-15(10-3-5-11(6-4-10)20(23)24)13-8-7-12(21)9-14(13)26-17(16)19/h3-9,15,21H,2,19H2,1H3
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| Chemical Name |
ethyl 2-amino-7-hydroxy-4-(4-nitrophenyl)-4H-chromene-3-carboxylate
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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) |
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 | 2.8064 mL | 14.0319 mL | 28.0639 mL | |
| 5 mM | 0.5613 mL | 2.8064 mL | 5.6128 mL | |
| 10 mM | 0.2806 mL | 1.4032 mL | 2.8064 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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