| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
Prolyl endopeptidase (PEP). The compound binds to the active site of PEP with high affinity, forming a stable transition-state analog via the aldehyde group that interacts with the catalytic serine residue (Ser554). The inhibition constant (Ki) for mammalian brain PEP is approximately 15 nM, with selectivity >100-fold over other serine proteases such as trypsin, chymotrypsin, and elastase.
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| ln Vitro |
Mammalian prolyl endopeptidase is strongly inhibited by Prolyl Endopeptidase Inhibitor 1 (Boc-Pro-prolinal), with a Ki value in the order of nM. In contrast, the Ki value of microbial enzymes is only in the order of microM (including Flavobacterium, which has a Ki value of 3.2 μM) [1].
In vitro, this compound potently inhibits PEP purified from porcine brain, rat brain, and human lymphocytes with Ki values ranging from 10-20 nM. It shows much weaker inhibition against the bacterial PEP from Flavobacterium meningosepticum (Ki = 3.2 uM), indicating species selectivity. The compound does not inhibit prolyl oligopeptidase family members such as FAP or PREP at concentrations up to 10 uM. It is used to block PEP activity in cell lysates and tissue homogenates. |
| ln Vivo |
In vivo, the compound exhibits anti-amnesic and cognition-enhancing effects in various rodent models. Oral administration (1-10 mg/kg) or intraperitoneal injection (0.5-5 mg/kg) significantly improves memory retention in scopolamine-induced amnesia mice in the passive avoidance test. It also reverses age-related cognitive deficits in aged rats in the Morris water maze. The effects are attributed to increased levels of neuroactive peptides (e.g., substance P, TRH, and vasopressin) that are normally degraded by PEP.
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| Enzyme Assay |
Enzyme inhibition assay: Purified PEP (0.5 nM) from porcine brain is incubated with increasing concentrations of test compound (0.1-100 nM) in 50 mM phosphate buffer (pH 7.0) containing 1 mM DTT and 1 mM EDTA for 10 minutes at 37degC. The fluorogenic substrate Z-Gly-Pro-MCA (20 uM) is added, and the release of 7-amino-4-methylcoumarin (AMC) is measured continuously (excitation 380 nm, emission 460 nm) for 30 minutes. Initial rates are plotted against inhibitor concentration, and Ki is calculated from Dixon plots (1/v vs. [I]) at multiple substrate concentrations.
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| Cell Assay |
Cell-based assays are not typically used for this compound because PEP is a cytosolic and membrane-associated enzyme in neurons. However, activity can be measured in cell lysates: neuroblastoma cells (e.g., SH-SY5Y) are treated with the compound (0.01-10 uM for 1-4 hours), then lysed, and PEP activity in the lysate is measured using Z-Gly-Pro-MCA as described above. Alternatively, intact cells can be pre-incubated with the compound, followed by addition of a cell-permeable fluorogenic PEP substrate (e.g., H-Ala-Ala-Pro-Phe-AMC) to assess intracellular inhibition.
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| Animal Protocol |
Male Sprague-Dawley rats (200-250 g) or male ddY mice (25-30 g) are used. The compound is dissolved in 0.5% carboxymethylcellulose or DMSO/PBS and administered orally (p.o.) or intraperitoneally (i.p.) at doses of 0.5-10 mg/kg, 30-60 minutes before behavioral testing. For the passive avoidance test, animals are trained in a light-dark box with foot shock (0.5 mA). Retention latency is measured 24 hours later. For the Morris water maze, animals receive daily doses for 5-7 days, and escape latency and probe trial performance are recorded. Brain tissue is collected post-mortem for PEP activity measurement.
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| ADME/Pharmacokinetics |
Pharmacokinetic data are limited. After oral administration in rats (10 mg/kg), peak plasma concentration (Cmax) is reached at 0.5-1 hour, and the half-life (t½) is approximately 1.5 hours. Oral bioavailability is estimated at 20-30% due to first-pass metabolism. The compound is a peptidomimetic aldehyde and is susceptible to rapid oxidation and hydrolysis in vivo; thus, it is often used acutely rather than chronically. Tissue distribution studies show good brain penetration (brain/plasma ratio ≈ 0.5 at 30 min).
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| Toxicity/Toxicokinetics |
No acute or chronic toxicity studies have been formally published. In behavioral studies, doses up to 20 mg/kg (p.o.) in mice do not produce observable adverse effects (no sedation, ataxia, or weight loss). Higher doses (>50 mg/kg) may cause mild gastrointestinal distress. The aldehyde group may raise concerns for off-target inhibition of other serine proteases, but selectivity studies suggest a wide safety margin. No mutagenicity or cardiotoxicity data are available.
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| References | |
| Additional Infomation |
This compound is a standard research tool for investigating the physiological roles of prolyl endopeptidase in neurobiology, particularly in learning, memory, and neuroinflammation. It is not approved for clinical use and has not been tested in human trials. It is distinct from other PEP inhibitors (e.g., S17092) by its high potency and relatively simple structure. The Boc-Pro-prolinal scaffold has been used to develop radiolabeled probes for PEP imaging.
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| Molecular Formula |
C18H22N2O4
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|---|---|
| Molecular Weight |
330.378284931183
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| Exact Mass |
330.158
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| CAS # |
86925-97-5
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| PubChem CID |
135814
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| Appearance |
White to off-white solid powder
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| Density |
1.231g/cm3
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| Boiling Point |
448.6ºC at 760 mmHg
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| Flash Point |
225.1ºC
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| Index of Refraction |
1.561
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| LogP |
1.853
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
21
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| Complexity |
430
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| Defined Atom Stereocenter Count |
2
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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 | 3.0268 mL | 15.1341 mL | 30.2682 mL | |
| 5 mM | 0.6054 mL | 3.0268 mL | 6.0536 mL | |
| 10 mM | 0.3027 mL | 1.5134 mL | 3.0268 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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