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Leupeptin, Microbial

In stock
Catalog No.
A2570
Inhibitor of serine and cysteine proteases
Grouped product items
SizePriceStock Qty
25mg
$100.00
In stock

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Background

Leupeptin is a reversible inhibitor of protease with Ki values of 35 nM, 3.4 μM, 6 nM and 72 nM for bovine trypsin, human plasmin, bovine spleen cathepsin B and recombinant human calpain, respectively [1, 2].

As a protease inhibitor, leupeptin was originally isolated from the Streptomyces species. It exerted poor membrane permeability due to its polar C-terminal. For calpain, leupeptin showed moderate potent activities with IC50 values of 0.211 μM, 1.8 μM and 0.938 μM against the enzymes isolated from porcine erythrocyte, porcine kidney and human platelet, respectively. In cultured MRC-C cells, leupeptin suppressed the growth of human coronavirus strain 229E through inhibited the activity of trypsin. The mean IC50 value was 0.8 μM [2, 3].

References:
1.  Mehdi S. Cell-penetrating inhibitors of calpain. Trends in biochemical sciences, 1991, 16: 150-153.
2.  Krauser J A, Powers J C. 6.1 BIOLOGICAL ROLES. Proteinase and Peptidase Inhibition: Recent Potential Targets for Drug Development, 2003: 144.
3.  Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. Journal of general virology, 1985, 66(2): 363-366.

Chemical Properties

StorageStore at -20°C
The product is not stable in solution, please dissolve it immediately before use.
M.Wt 475.59
Cas No.103476-89-7
FormulaC20H38N6O4.1/2H2SO4
SynonymsLeupeptin hemisulfate salt microbial,L-Leucinamide,Leupeptin, Microbial
Solubility≥24.7mg/mL in DMSO
Chemical Name2-acetamido-N-(1-((5-((diaminomethylene)amino)-1-oxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)-4-methylpentanamide
SDFDownload SDF
Canonical SMILESO=C(C(C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H])([H])N([H])C(C([H])([H])[H])=O)N([H])C(C(N([H])C(C([H])=O)([H])C([H])([H])C([H])([H])C([H])([H])/N=C(N([H])[H])/N([H])[H])=O)([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H]
Shipping ConditionEvaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request
General tipsFor obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.

Protocol

Cell experiment [1]:

Cell lines

MRC-C cells infected with HCV 229E

Preparation method

The solubility of this compound in DMSO is ≥49.35mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

0, 1, 10 and 100 μg/mL; 24 hrs

Applications

In cultures of MRC-C cells, Leupeptin prevented multiplication of the human coronavirus strain 229E. The IC50 value of Leupeptin in plaque tests was 0.4 μg/mL, whilst growth of host cells was unaffected by Leupeptin at 50 μg/mL. In single-cycle growth experiments, Leupeptin (100 μg/mL) reduced virus yield only if added within 2 hrs of infection, indicating its action on an early stage of virus replication.

Animal experiment [2]:

Animal models

C57BL/6NCrl male mice

Dosage form

0, 9, 18 36 and 40 mg/kg; i.p.

Applications

Leupeptin was well tolerated by the animals and dose-dependently produced a substantial increase in LC3b-II in both the total tissue extracts and the lysosome enriched fraction (LE fraction). At the electron microscopy (EM) level, leupeptin induced the accumulation of electron-dense vesicular structures that, in hepatocytes, were visible by 60 min after treatment (40 mg/kg). The results suggested that Leupeptin enhanced LC3b-II levels in vivo by protecting this protein from being degraded inside lysosomes, and thus the leupeptin-based assay could be potentially used for studying the dynamics of macroautophagy in mice.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. Journal of general virology, 1985, 66(2): 363-366.

[2]. Haspel J, Shaik RS, Ifedigbo E, Nakahira K, Dolinay T, Englert JA, Choi AM. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42.

Biological Activity

Description Leupeptin, Microbial(Leupeptin hemisulfate ) is an reversible, competitive inhibitor of the proteases with Ki values of 0.13 nM and 7 nM for trypsin and cathepsin B, respectively.
Targets Trypsin Cathepsin B        
IC50 0.13 nM (Ki) 7 nM (Ki)        

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