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E-64

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Catalog No.
A2576
半胱氨酸蛋白酶(cysteine protease)抑制剂
组合的产品项目
规格价格库存 数量
10mM (in 1mL DMSO)
¥ 550.00
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5mg
¥ 500.00
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25mg
¥ 1,400.00
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100mg
¥ 4,200.00
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250mg
¥ 9,800.00
现货

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Background

A new class of compounds that show promise of acting as class-specific inhibitors for the cysteine proteinases are the L-trans-epoxysuccinylpeptides related to the compound E-64 [L-trans-epoxysuccinyl-L-leucylamido(4-guanidino)butanel , isolated from cultures of Aspergillus. E-64 was shown to inhibit papain, ficin and the fruit and stem bromelains, with disappearance of the thiol group of papain1.

 E-64 has been reported to inhibit two other mammalian cysteine proteinases: cathepsin L3 and a proteinase from human breast-tumour tissue4 and the calcium-dependent proteinase, calpain, from chicken muscle5. All of these characteristics suggested that E-64 might be a valuable inhibitor for the study of cysteine proteinases.

Lineweaver-Burk plots of inhibition data show that the action of E-64 was not competitive with substrate1 . Moreover, the optical isomerism of the epoxysuccinyl moiety seemed to have no effect on the activity of E-64 as an inhibitor of papain6, 7 .If E-64 were indeed acting by covalent reaction at the active site, its rate of reaction would be decreased by the presence of leupeptin, a tight-binding reversible inhibitor8.

E-64 inhibits only cysteine proteinases. Papain showed a particularly high reactivity with E-64, and good rates were also obtained with the other plant enzymes and the lysosomal cysteine proteinases. There is structural evidence that these enzymes form a homologous group9, and they resemble each other in having Mr about 25 000, no (detected) zymogens and no distinct requirement for calcium. Chicken skeletal-muscle calpain is reported to be inhibited by E-64, but the rate constant has not been determined5.

The most obvious practical application of E-64 is in the active-site titration of the papain-related cysteine proteinases. Active-site titration as a method of determining enzyme concentration has the advantage over rate assays of being insensitive to reaction conditions, and giving a result in active-site molarity10 (Bender et al., 1966).

References:
1. A. J. BARRETT, A. A. KEMBHAVI, L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) and its analogues as inhibitors of cysteine proteinases including cathepsins B, H and L. Biochem. J. (1982) 201, 189-198
2. Hanada, K., Tamai, M., Yamagishi, M., Ohmura, S., Sawada, J. & Tanaka, I. (1978c) Agric. Biol. Chem. 42, 523-528
3. Towatari, T., Tanaka, K., Yoshikawa, D. & Katunuma, N. (1978).J. Biochem. (Tokyo) 84, 659-671.
4. Mort, J. S., Recklies, A. D. & Poole, A. R. (1980) Biochim. Biophys. Acta 614, 134-143.
5. Sugita, H., Ishiura, S., Suzuki, K. & Imahori, K. (1980) J. Biochem. (Tokyo) 87, 339-341
6. Hanada, K., Tamai, M., Morimoto, S., Adachi, T.,Ohmura, S., Sawada, J. & Tanaka, I. (1978a) Agric. Biol. Chem. 42, 537-541.
7. Hanada, K., Tamai, M., Ohmura, S., Sawada, J., Seki, T.& Tanaka, I. (1978b)Agric. Biol. Chem. 42, 529-536
8. Knight, C. G. (1980) Biochem. J. 189,447-453
9. Takio, K., Towatari, T., Katunuma, N. & Titani, K.(1980) Biochem. Biophys. Res. Commun. 97, 340-346
10. Bender, M. L., Begue-Canton, M. L., Blakeley, R. L.,Brubacher, L. J., Feder, J., Gunter, C. R., Kezdy, F. J.,Killheffer, J. V., Marshall, T. H., Miller, C. G., Roeske,R. W. & Stoops, J. K. (1966) J. Am. Chem. Soc. 88,5890-5913

文献引用

1. Blass G, Levchenko V, et al. "Chronic cathepsin inhibition by E-64 in Dahl salt-sensitive rats." Physiol Rep. 2016 Sep;4(17). pii: e12950. PMID:27597769
2. Ying Long, Xuri Zhang, et al. "Initial events in the breakthrough of the epithelial barrier of the small intestine by Angiostrongylus cantonensis." Arch Biol Sci. 2016;68(2):375-383

Chemical Properties

Physical AppearanceA solid
StorageStore at 2-8°C
M.Wt357.41
Cas No.66701-25-5
FormulaC15H27N5O5
Solubility≥53.6mg/mL in DMSO
Chemical Name(2S,3S)-3-[[(2S)-1-[4-(diaminomethylideneamino)butylamino]-4-methyl-1-oxopentan-2-yl]carbamoyl]oxirane-2-carboxylic acid
SDFDownload SDF
Canonical SMILESCC(C)CC(C(=O)NCCCCN=C(N)N)NC(=O)C1C(O1)C(=O)O
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

细胞实验[1]:

细胞系

H-59和M-27细胞

溶解方法

在DMSO中的溶解度≥53.6mg/mL。为了获得更高的浓度,可以将离心管在37℃加热10分钟和/或在超声波浴中震荡一段时间。原液可以在-20℃以下储存几个月。

反应条件

10 μg/ml;48 h

应用

E-64以剂量依赖的方式抑制H-59细胞的侵袭,在10 μg/ml浓度时达到97%的最大抑制,是无毒性的。用包被7.5 μg/滤波器IV型胶原的过滤器测得细胞迁移仅减少25%,表明在缺乏基底膜屏障时,半胱氨酸蛋白酶在细胞迁移中发挥更加次要的作用。然而,E-64对M-27细胞的侵袭没有显著影响,甚至在高达100 μg/ml时。

动物实验[2]:

动物模型

Wistar系大鼠

剂量

1 mg/100 g体重;腹腔注射

应用

在注射E-64后1小时杀死动物,测定溶酶体中cathepsin B和cathepsin L的活性。结果表明,E-64抑制cathepsin B和cathepsin L的活性。

注意事项

请测试所有化合物在室内的溶解度,实际溶解度和理论值可能略有不同。这是由实验系统的误差引起的,属于正常现象。

References:

[1] Navab R, Mort J S, Brodt P. Inhibition of carcinoma cell invasion and liver metastases formation by the cysteine proteinase inhibitor E-64. Clinical & experimental metastasis, 1997, 15(2): 121-129.

[2] Hashida S, TOWATARI T, KOMINAMI E, et al. Inhibitions by E-64 derivatives of rat liver cathepsin B and cathepsin L in vitro and in vivo. Journal of biochemistry, 1980, 88(6): 1805-1811.

生物活性

E-64是一种天然的、有效的和不可逆的半胱氨酸蛋白酶抑制剂,作用于cathepsins K、S和L,IC50值分别为1.4、4.1和2.5 nM。.
靶点 cathepsins K cathepsins S cathepsins L      
IC50 1.4nM 4.1nM 2.5nM      

质量控制

质量控制和MSDS

批次:

化学结构

E-64

相关生物数据

E-64

相关生物数据

E-64

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E-64

相关生物数据

E-64