CAS NO: | 66701-25-5 |
规格: | ≥98% |
包装 | 价格(元) |
5mg | 电议 |
10mg | 电议 |
25mg | 电议 |
50mg | 电议 |
100mg | 电议 |
250mg | 电议 |
500mg | 电议 |
Molecular Weight (MW) | 393.86 |
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Formula | C15H28CLN5O5 |
CAS No. | N/A; 66701-25-5 (free base) |
Storage | -20℃ for 3 years in powder form |
-80℃ for 2 years in solvent | |
Solubility (In vitro) | DMSO: 71 mg/mL (198.7 mM) |
Water: <1 mg/mL | |
Ethanol: 11 mg/mL (30.8 mM) | |
SMILES Code | O=C([C@@H]1[C@@H](C(O)=O)O1)N[C@H](C(NCCCCNC(N)=N)=O)CC(C)C.[H]Cl |
Synonyms | E-64 HCl; E 64 HCl; E64 HCl |
In Vitro | In vitro activity: 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. E-64, as a specific inhibitor os cysteine proteases, effectively blocks in vitro ovarian cancer cell invasion. In addition, E-64 also shows antiparasitic activity against Giardia lamblia excystation in vitro. E-64 improves the preimplantation development of bovine somatic cell nuclear transfer embryos, which indicates another important implication of E-64. Kinase Assay: The Cathepsin B 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. Cell Assay: E-64 inhibited H-59 invasion in a dose-dependent manner with a maximal inhibition of 97% at a concentration of 10 μg/ml which was non-toxic. Cell migration as measured with filters coated with 7.5 μg/filter type IV collagen was reduced by only 25% suggesting that the cysteine proteinases played a more minor role in cell migration in the absence of a basement membrane barrier. On the other hand M-27 invasion was not significantly affected by treatment with E-64 even at concentrations as high as 100 μg/ml. |
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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. E-64 also shows antiparasitic activity against Giardia lamblia excystation in infected mice. |
Animal model | Wistar strain rats |
Formulation & Dosage | 1 mg/100 g body weight; i.p. injection |
References | Biopolymers. 1999;51(1):99-107; J Egypt Soc Parasitol. 2009 Apr;39(1):111-9; J Biochem. 1982 Apr;91(4):1373-80. |