WARNING: This product is for research use only, not for human or veterinary use.
Hodoodo CAT#: H530905
CAS#: 66701-25-5
Description: E-64 is a potent, and irreversible inhibitor of cysteine proteases with IC50 values for inhibiting cathepsins K, S, and L, in vitro, are 1.4, 4.1, and 2.5 nM, respectively. E-64 improves the preimplantation development of bovine somatic cell nuclear transfer embryos.
Hodoodo Cat#: H530905
Name: E-64
CAS#: 66701-25-5
Chemical Formula: C15H27N5O5
Exact Mass: 357.20
Molecular Weight: 357.411
Elemental Analysis: C, 50.41; H, 7.61; N, 19.60; O, 22.38
Synonym: E-64; E 64; E64.
IUPAC/Chemical Name: 3-[[[(1S)-1-[[[4-[(aminoiminomethyl)amino]butyl]amino]carbonyl]-3-methylbutyl]amino]carbonyl]-(2S,3S)-oxiranecarboxylic acid
InChi Key: LTLYEAJONXGNFG-DCAQKATOSA-N
InChi Code: InChI=1S/C15H27N5O5/c1-8(2)7-9(20-13(22)10-11(25-10)14(23)24)12(21)18-5-3-4-6-19-15(16)17/h8-11H,3-7H2,1-2H3,(H,18,21)(H,20,22)(H,23,24)(H4,16,17,19)/t9-,10-,11-/m0/s1
SMILES Code: O=C([C@@H]1[C@@H](C(O)=O)O1)N[C@H](C(NCCCCNC(N)=N)=O)CC(C)C
Appearance: Solid powder
Purity: >98% (or refer to the Certificate of Analysis)
Shipping Condition: Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition: Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility: Soluble in DMSO
Shelf Life: >2 years if stored properly
Drug Formulation: This drug may be formulated in DMSO
Stock Solution Storage: 0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code: 2934.99.9001
More Info:
| Biological target: | E-64 (Proteinase inhibitor E 64) is a potent irreversible inhibitor against general cysteine proteases with IC50 of 9 nM for papain. |
| In vitro activity: | E-64 showed a concentration and time dependent decrease in motility and viability of the parasites as shown in Table 1 and Figure 1A respectively (EC50 = 16 μM). The microfilariae (mf) recovered from the uterus of gravid female parasites after 8 h were found to be motile up to 10 μM. They were under extreme stress and dead in case of 20 μM and 40 μM E-64 treated parasites at the end of 8 h (Figure 1B). Reference: PLoS One. 2014; 9(3): e93161. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3965533/ |
| In vivo activity: | A significant increase in the renal cortical mature form of Cath B (Fig. 7A) and Cath L (Fig. 7B) were measured in E‐64 treated rats (P < 0.01, independent‐sample t‐test). The pro Cath L abundance was not different between the groups and pro Cath B was not detected. Reference: Physiol Rep. 2016 Sep; 4(17): e12950. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5027357/ |
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 48.7 | 136.17 | |
| Water | 15.9 | 44.51 |
The following data is based on the product molecular weight 357.41 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 1 mM | 1.15 mL | 5.76 mL | 11.51 mL |
| 5 mM | 0.23 mL | 1.15 mL | 2.3 mL |
| 10 mM | 0.12 mL | 0.58 mL | 1.15 mL |
| 50 mM | 0.02 mL | 0.12 mL | 0.23 mL |
| Formulation protocol: | 1. Stoute JA, Landmesser ME, Biryukov S. Treatment of Plasmodium falciparum merozoites with the protease inhibitor E64 and mechanical filtration increases their susceptibility to complement activation. PLoS One. 2020 Aug 21;15(8):e0237786. doi: 10.1371/journal.pone.0237786. PMID: 32822376; PMCID: PMC7442247. 2. Wadhawan M, Singh N, Rathaur S. Inhibition of cathepsin B by E-64 induces oxidative stress and apoptosis in filarial parasite. PLoS One. 2014 Mar 25;9(3):e93161. doi: 10.1371/journal.pone.0093161. PMID: 24667798; PMCID: PMC3965533. 3. Blass G, Levchenko V, Ilatovskaya DV, Staruschenko A. Chronic cathepsin inhibition by E-64 in Dahl salt-sensitive rats. Physiol Rep. 2016 Sep;4(17):e12950. doi: 10.14814/phy2.12950. PMID: 27597769; PMCID: PMC5027357. 4. Hussein EM, Dawoud HA, Salem AM, Atwa MM. Antiparasitic activity of cystine protease inhibitor E-64 against Giardia lamblia excystation in vitro and in vivo. J Egypt Soc Parasitol. 2009 Apr;39(1):111-9. PMID: 19530614. |
| In vitro protocol: | 1. Stoute JA, Landmesser ME, Biryukov S. Treatment of Plasmodium falciparum merozoites with the protease inhibitor E64 and mechanical filtration increases their susceptibility to complement activation. PLoS One. 2020 Aug 21;15(8):e0237786. doi: 10.1371/journal.pone.0237786. PMID: 32822376; PMCID: PMC7442247. 2. Wadhawan M, Singh N, Rathaur S. Inhibition of cathepsin B by E-64 induces oxidative stress and apoptosis in filarial parasite. PLoS One. 2014 Mar 25;9(3):e93161. doi: 10.1371/journal.pone.0093161. PMID: 24667798; PMCID: PMC3965533. |
| In vivo protocol: | 1. Blass G, Levchenko V, Ilatovskaya DV, Staruschenko A. Chronic cathepsin inhibition by E-64 in Dahl salt-sensitive rats. Physiol Rep. 2016 Sep;4(17):e12950. doi: 10.14814/phy2.12950. PMID: 27597769; PMCID: PMC5027357. 2. Hussein EM, Dawoud HA, Salem AM, Atwa MM. Antiparasitic activity of cystine protease inhibitor E-64 against Giardia lamblia excystation in vitro and in vivo. J Egypt Soc Parasitol. 2009 Apr;39(1):111-9. PMID: 19530614. |
1: Blass G, Levchenko V, Ilatovskaya DV, Staruschenko A. Chronic cathepsin inhibition by E-64 in Dahl salt-sensitive rats. Physiol Rep. 2016 Sep;4(17). pii: e12950. doi: 10.14814/phy2.12950. PubMed PMID: 27597769; PubMed Central PMCID: PMC5027357.
2: Wilder CL, Walton C, Watson V, Stewart FA, Johnson J, Peyton SR, Payne CK, Odero-Marah V, Platt MO. Differential cathepsin responses to inhibitor-induced feedback: E-64 and cystatin C elevate active cathepsin S and suppress active cathepsin L in breast cancer cells. Int J Biochem Cell Biol. 2016 Oct;79:199-208. doi: 10.1016/j.biocel.2016.08.030. Epub 2016 Aug 31. PubMed PMID: 27592448; PubMed Central PMCID: PMC5067213.
3: Kershaw CM, Evans G, Rodney R, Maxwell WM. Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates. Reprod Fertil Dev. 2016 May 9. doi: 10.1071/RD15261. [Epub ahead of print] PubMed PMID: 27156102.
4: Zhang Z, Huang Z, Dai H, Wei L, Sun S, Gao F. Therapeutic Efficacy of E-64-d, a Selective Calpain Inhibitor, in Experimental Acute Spinal Cord Injury. Biomed Res Int. 2015;2015:134242. doi: 10.1155/2015/134242. Epub 2015 Jul 9. PubMed PMID: 26240815; PubMed Central PMCID: PMC4512559.
5: Wadhawan M, Singh N, Rathaur S. Inhibition of cathepsin B by E-64 induces oxidative stress and apoptosis in filarial parasite. PLoS One. 2014 Mar 25;9(3):e93161. doi: 10.1371/journal.pone.0093161. eCollection 2014. PubMed PMID: 24667798; PubMed Central PMCID: PMC3965533.
6: Grozdanovic MM, Ostojic S, Aleksic I, Andjelkovic U, Petersen A, Gavrovic-Jankulovic M. Active actinidin retains function upon gastro-intestinal digestion and is more thermostable than the E-64-inhibited counterpart. J Sci Food Agric. 2014 Nov;94(14):3046-52. doi: 10.1002/jsfa.6656. Epub 2014 Apr 10. PubMed PMID: 24633720.
7: Carvalho TB, Oliveira-Sequeira TC, Guimarães S. In vitro antigiardial activity of the cysteine protease inhibitor E-64. Rev Inst Med Trop Sao Paulo. 2014 Jan-Feb;56(1):43-7. doi: 10.1590/S0036-46652014000100006. PubMed PMID: 24553607; PubMed Central PMCID: PMC4085827.
8: Min SH, Song BS, Yeon JY, Kim JW, Bae JH, Park SY, Lee YH, Chang KT, Koo DB. A cathepsin B inhibitor, E-64, improves the preimplantation development of bovine somatic cell nuclear transfer embryos. J Reprod Dev. 2014 Mar 7;60(1):21-7. Epub 2013 Nov 16. PubMed PMID: 24240170; PubMed Central PMCID: PMC3963301.
9: Tanabe F, Nakajima T, Ito M. The thiol proteinase inhibitor E-64-d ameliorates amyloid-β-induced reduction of sAPPα secretion by reversing ceramide-induced protein kinase C down-regulation in SH-SY5Y neuroblastoma cells. Biochem Biophys Res Commun. 2013 Nov 8;441(1):256-61. doi: 10.1016/j.bbrc.2013.10.045. Epub 2013 Oct 17. PubMed PMID: 24141119.
10: Yang WX, Zhang WH, Wu SY, Yao K, Liang GB, Li Y. [Effect of different molarity cathepsins specific inhibitor E-64 on dentin-resin bonding durability]. Zhonghua Kou Qiang Yi Xue Za Zhi. 2013 Jun;48(6):368-71. Chinese. PubMed PMID: 24120008.
11: Grozdanović MM, Drakulić BJ, Gavrović-Jankulović M. Conformational mobility of active and E-64-inhibited actinidin. Biochim Biophys Acta. 2013 Oct;1830(10):4790-9. doi: 10.1016/j.bbagen.2013.06.015. Epub 2013 Jun 23. PubMed PMID: 23803410.
12: Hussein EM, Dawoud HA, Salem AM, Atwa MM. Antiparasitic activity of cystine protease inhibitor E-64 against Giardia lamblia excystation in vitro and in vivo. J Egypt Soc Parasitol. 2009 Apr;39(1):111-9. PubMed PMID: 19530614.
13: Tanabe F, Kasai H, He L, Kin T, Fujikado T, Kumamoto T, Hara T, Iwata T, Ito M. Improvement of deficient natural killer activity and delayed bactericidal activity by a thiol proteinase inhibitor, E-64-d, in leukocytes from Chediak-Higashi syndrome patients in vitro. Int Immunopharmacol. 2009 Mar;9(3):366-70. doi: 10.1016/j.intimp.2009.01.003. Epub 2009 Jan 29. PubMed PMID: 19185618.
14: Morimoto M, Tanabe F, Kasai H, Ito M. Effect of a thiol proteinase inhibitor, E-64-d, on susceptibility to infection with Staphylococcus aureus in Chediak-Higashi syndrome (beige) mice. Int Immunopharmacol. 2007 Jul;7(7):973-80. Epub 2007 Apr 16. PubMed PMID: 17499200.
15: Gavira JA, González-Ramírez LA, Oliver-Salvador MC, Soriano-García M, García-Ruiz JM. Structure of the mexicain-E-64 complex and comparison with other cysteine proteases of the papain family. Acta Crystallogr D Biol Crystallogr. 2007 May;63(Pt 5):555-63. Epub 2007 Apr 21. PubMed PMID: 17452780.
16: Yoshifuji H, Umehara H, Maruyama H, Itoh M, Tanaka M, Kawabata D, Fujii T, Mimori T. Amelioration of experimental arthritis by a calpain-inhibitory compound: regulation of cytokine production by E-64-d in vivo and in vitro. Int Immunol. 2005 Oct;17(10):1327-36. PubMed PMID: 16176933.
17: Chatterjee PK, Todorovic Z, Sivarajah A, Mota-Filipe H, Brown PA, Stewart KN, Mazzon E, Cuzzocrea S, Thiemermann C. Inhibitors of calpain activation (PD150606 and E-64) and renal ischemia-reperfusion injury. Biochem Pharmacol. 2005 Apr 1;69(7):1121-31. PubMed PMID: 15763548.
18: Sarabia F, Sánchez-Ruiz A, Chammaa S. Stereoselective synthesis of E-64 and related cysteine proteases inhibitors from 2,3-epoxyamides. Bioorg Med Chem. 2005 Mar 1;13(5):1691-705. PubMed PMID: 15698787.
19: Yoshidome K, Kobae H, Yamamoto K, Sameshima K, Miyata K, Kawano Y. Effects of supplemental L-methionine on E-64 [trans-epoxysuccinyl-1-leucyl-amido (4-guanido) butane]-induced dysmorphology in rat embryos cultured in vitro. Congenit Anom (Kyoto). 2003 Dec;43(4):280-5. PubMed PMID: 15041779.
20: Takahashi S, Zhang HC, Hasegawa R, Karata K, Kita K, Wano C, Yamaguchi Y, Sugaya S, Nomura J, Ichinose M, Suzuki N. Increased levels of UV-induced protease activity in human UVAP-1 cells exposed to gravity-changing stress: involvement of E-64-sensitive proteases in suppression of UV mutagenicity. Cell Biol Int. 2003;27(1):53-60. PubMed PMID: 12713800.
