| Name | E-64 |
| Description | E-64 (Proteinase inhibitor E 64) is an irreversible and specific inhibitor of cysteine proteases, exhibiting an IC50 of 9 nM for papain. |
| Kinase Assay | The Cathepsin Β activity is determined using Z-Arg-Arg-4mβNA as substrate with slight modifications. The crude extract is pre-incubated at 37°C for 5 min in 50 mM sodium acetate buffer, pH 5.0 containing 1 mM EDTA and 5 mM DTT. The substrate (final concentration, 100 μM) is added to make the final assay volume of 1 mL. The reaction mixture is incubated at 37°C for 30 min. The reaction is terminated by adding equal volume of stopping reagent containing Fast Garnet GBC salt (1 mg/mL), 10 mM pHMB and 50 mM EDTA, pH 6.0. The extraction of product, β-napthylamine (β-NA), is carried out with n-butanol. After complete layer separation, the absorbance is measured in n-butanol layer and activity is calculated using molar extinction coefficient of β-napthylamine solution as 31.5 M/cm per sec at 520 nm. One unit of enzyme activity is defined as the amount of enzyme liberating 1 μmol of βNA per minute at 37°C. The half maximal inhibitory concentration (IC50) is calculated by plotting the graph between the different concentration of E-64 and the % inhibition in cathepsin B activity. Here, IC50 indicates the concentration of the E-64 required to inhibit the parasitic cathepsin B activity by half[2]. |
| In vitro | E-64 rapidly inhibits the activities of the cysteine proteinases cathepsins B, H and L and papain, while has no effect on the serine proteinases or the metalloproteinases. [2] E-64, as a specific inhibitor os cysteine proteases, effectively blocks in vitro ovarian cancer cell invasion. [3] In addition, E-64 also shows antiparasitic activity against Giardia lamblia excystation in vitro. [4] E-64 improves the preimplantation development of bovine somatic cell nuclear transfer embryos, which indicates another important implication of E-64. [5] |
| In vivo | E-64 induces a marked decrease in the number of spontaneous metastasis in M5076 tumor bearing mice, while has no effect on the formation of experimental metastasis. [6] E-64 also shows antiparasitic activity against Giardia lamblia excystation in infected mice. [4] |
| Storage | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| Solubility Information | H2O : 9 mg/mL (25.2 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 50 mg/mL (139.9 mM)
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| Keywords | Cathepsin | Inhibitor | Proteinase inhibitor E-64 | inhibit | Proteinase inhibitor E64 | E-64 | Bacterial | Autophagy |
| Inhibitors Related | Neomycin sulfate | Stavudine | Ampicillin sodium | Kanamycin sulfate | Sodium 4-phenylbutyrate | Sulfamethoxazole sodium | Hydroxychloroquine | Guanidine hydrochloride | Doxycycline | Paeonol | Naringin | Dimethyl sulfoxide |
| Related Compound Libraries | Apoptosis Compound Library | Bioactive Compound Library | Autophagy Compound Library | Inhibitor Library | NO PAINS Compound Library | Bioactive Compounds Library Max | Preclinical Compound Library | Anti-Cancer Compound Library | Anti-COVID-19 Compound Library | Anti-Cancer Active Compound Library |
United States
