| Size | Price | Stock | Qty |
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| Targets |
The compound targets calpain I (micro-calpain), a calcium-activated cysteine protease, with a Ki value of 19 nM, demonstrating high selectivity. It exhibits lower affinity for calpain II (m-calpain, Ki = 120 nM) and cathepsin B (Ki = 750 nM). Calpain I is primarily involved in cellular signaling and cytoskeletal remodeling, and its dysregulation is implicated in numerous pathological processes, making this selective inhibitor a crucial tool for target validation.
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| ln Vitro |
In cell-free enzymatic assays, Calpain Inhibitor XII demonstrates reversible, selective inhibition of calpain I with a Ki value of 19 nM. Its IC50 is determined by measuring the reduction in calpain I-mediated proteolysis of a fluorogenic peptide substrate such as Suc-LLVY-AMC or casein. The compound shows significantly lower potency against calpain II and cathepsin B, confirming its selectivity for the micro-calpain isoform.
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| ln Vivo |
Specific in vivo data for this exact compound are not detailed in the literature; however, calpain inhibitors in general show efficacy in animal models of neurodegeneration (e.g., stroke, traumatic brain injury, Alzheimer's disease), cardiac ischemia-reperfusion injury, and inflammation. The compound's cell permeability and selectivity for calpain I over calpain II suggest it could effectively inhibit micro-calpain-mediated pathology without complete abrogation of m-calpain's physiological functions, potentially offering a favorable safety profile.
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| Enzyme Assay |
The typical biochemical assay for calpain I inhibition involves a fluorometric protease activity assay. Purified recombinant or tissue-extracted calpain I (micro-calpain) is incubated with a fluorogenic substrate, such as Suc-LLVY-AMC or casein labeled with a fluorophore, in a calcium-containing reaction buffer (e.g., 50 mM Tris-HCl, pH 7.5, 5 mM CaCl2, 10 mM 2-mercaptoethanol). Varying concentrations of Calpain Inhibitor XII are added, and the mixture is incubated at 37degC for 30-60 minutes. The increase in fluorescence (Ex/Em ~380/460 nm for AMC) is monitored continuously or after stopping the reaction. The IC50 or Ki is calculated from the dose-response curve.
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| Cell Assay |
A cellular assay involves treating cultured neurons or cardiomyocytes with Calpain Inhibitor XII prior to an apoptotic or necrotic stimulus (e.g., calcium ionophore, oxidative stress, or simulated ischemia). After treatment, cells are harvested, and the level of calpain-mediated proteolysis is assessed by Western blotting using antibodies against known calpain substrates such as alpha-spectrin (which is cleaved into characteristic 150 kDa and 145 kDa fragments), fodrin, or p35/p25. Alternatively, cell viability is measured using MTT or LDH release assays to assess the protective effect of calpain inhibition.
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| Animal Protocol |
In vivo animal experimental procedures for calpain inhibitors typically involve a mouse model of ischemia-reperfusion injury or traumatic brain injury. Mice are administered Calpain Inhibitor XII via intraperitoneal injection (e.g., at doses of 1-10 mg/kg) 30 minutes prior to the insult and possibly again post-insult. In a stroke model, middle cerebral artery occlusion (MCAO) is performed for 60-90 minutes followed by reperfusion. After 24-48 hours, neurological deficit scores are assessed, and brain tissues are harvested to measure infarct volume by TTC staining and calpain-mediated alpha-spectrin breakdown products by Western blot.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic parameters for Calpain Inhibitor XII are not detailed in available literature. As a small-molecule peptidomimetic inhibitor (MW 482.6), it is designed to be cell-permeable and is typically administered via intraperitoneal or intravenous injection in animal studies. Its stability in circulation, tissue distribution, and ability to cross the blood-brain barrier are critical factors for its use in neurological research, though these parameters are often compound-specific and would require empirical determination.
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| Toxicity/Toxicokinetics |
Specific toxicological data for Calpain Inhibitor XII are not detailed in the literature. As a selective inhibitor of calpain I, its safety profile is expected to be manageable, given that calpain I is primarily activated under pathological conditions rather than being constitutively active. However, chronic or high-dose inhibition could potentially interfere with normal calcium signaling and cellular homeostasis, including effects on cell migration, platelet function, and synaptic plasticity. Standard toxicological endpoints would be assessed in animal studies.
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| References |
:Sci Adv. 2020 Nov 6:eabe0751.
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| Additional Infomation |
Calpain Inhibitor XII is a research-grade chemical tool, not a clinical drug candidate. Calpains are involved in diverse pathological processes including neurodegeneration (Alzheimer's, Parkinson's, Huntington's diseases), muscular dystrophy, cardiac ischemia-reperfusion injury, cataract formation, and inflammation. This selective inhibitor allows researchers to dissect the specific contributions of calpain I versus calpain II in these conditions. As of the latest updates, the compound has not been approved for clinical use and is exclusively available for pre-clinical research purposes.
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| Molecular Formula |
C26H34N4O5
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|---|---|
| Molecular Weight |
482.572
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| Exact Mass |
482.252
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| CAS # |
181769-57-3
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| PubChem CID |
16760340
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| Appearance |
White to yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Index of Refraction |
1.548
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| LogP |
3.17
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
14
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| Heavy Atom Count |
35
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| Complexity |
693
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCC(C(=O)C(=O)NCC1=CC=CC=N1)NC(=O)C(CC(C)C)NC(=O)OCC2=CC=CC=C2
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| InChi Key |
PLVWMBFPIAQRHK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H34N4O5/c1-4-10-21(23(31)25(33)28-16-20-13-8-9-14-27-20)29-24(32)22(15-18(2)3)30-26(34)35-17-19-11-6-5-7-12-19/h5-9,11-14,18,21-22H,4,10,15-17H2,1-3H3,(H,28,33)(H,29,32)(H,30,34)
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| Chemical Name |
benzyl N-[1-[[1,2-dioxo-1-(pyridin-2-ylmethylamino)hexan-3-yl]amino]-4-methyl-1-oxopentan-2-yl]carbamate
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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 (e.g. under nitrogen), 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) |
DMSO : ~16 mg/mL (~33.16 mM)
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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 | 2.0722 mL | 10.3612 mL | 20.7224 mL | |
| 5 mM | 0.4144 mL | 2.0722 mL | 4.1445 mL | |
| 10 mM | 0.2072 mL | 1.0361 mL | 2.0722 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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