E64-RO

E-64

N-[N-(L-3-Trans-carboxirane-2-carbonyl)-L-leucyl]-agmatine

• Synonym(s): E-64, n-[n-( l-transepoxysuccinic acid)-l-leucyl]-agmatine, n-[n-(l-3-trans-carboxirane-2-carbonyl)-l-leucyl]-4-aminobutylguanidine, n-[n-(l-3-trans-carboxirane-2-carbonyl)-l-leucyl]-agmatine. ; note: another name for l-3-trans-carboxyoxiran-2-carboxylic acid acid is l-transepoxysuccinic acid. agmatine is 4-aminobutylguanidine., trans-Epoxysuccinyl-L-leucylamido(4-guanidino)butane, L-trans-3-Carboxyoxiran-2-carbonyl-L-leucylagmatine, N-(trans-Epoxysuccinyl)-L-leucine 4-guanidinobutylamide
• Empirical Formula (Hill Notation): C₁₅H₂₇N₅O₅
• CAS Number: 66701-25-5
• Molecular Weight: 357.41
• Beilstein: 1405664
• Numer MDL: MFCD00080261
• Identyfikator substancji w PubChem: 329798998

Properties

• Poziom jakości: 100
• próba: >95% (HPLC)
• forma: powder
• masa cząsteczkowa: M_r 357.4
• opakowanie: pkg of 10 mg (10874523001); pkg of 25 mg (11585681001)
• manufacturer/tradename: Roche
• pH range: 2-10
• rozpuszczalność: ethanol: water (1:1): soluble 20 mg/mL
• temp. przechowywania: 2-8°C
• SMILES string: CC(C)C[C@H](NC(=O)[C@@H]1O[C@H]1C(O)=O)C(=O)NCCCCNC(N)=N
• 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
• InChI key: LTLYEAJONXGNFG-HBNTYKKESA-N

Description

Specyficzność

The thiol protease inhibitor E-64 specifically inhibits papain and other cysteine proteases such as cathepsin B and L, bromelain, and ficin. Cathepsin A and D are not inhibited. L-lactate dehydrogenase from porcine heart is not inhibited, although the enzyme contains a functional thiol group.

Zastosowanie

E-64 is an effective ligand for affinity purification of cysteine proteases. When coupled to a thiolated affinity matrix, binding is no longer irreversible, but specificity is retained.

E-64 is used for the inhibition of papain and other cysteine proteases such as cathepsin B and L, bromelain, and ficin during isolation and purification of proteins and enzymes.
It has been used in the affinity chromatography for the purification of hemoglobin receptors.[1]
The inhibition of thiol proteases by E-64 appears to be of a non-competitive nature between the SH components. The inhibition is also irreversible, and is altered after gel filtration (Sephadex column) or dialysis after incubation of papain with E-64. The enzyme and inhibitor combine in an equimolar ratio.

Działania biochem./fizjol.

E-64 is an irreversible, potent, and highly selective cysteine protease inhibitor. E-64 does not react with the functional thiol group of non-protease enzymes, such as L-lactate dehydrogenase or creatine kinase. E-64 will not inhibit serine proteases (except trypsin) like other cysteine protease inhibitors, leupeptin and antipain. The trans-epoxysuccinyl group (active moiety) of E-64 irreversibly binds to an active thiol group in many cysteine proteases, such as papain, actinidase, and cathepsins B, H, and L to form a thioether linkage. E-64 is a very useful cysteine protease inhibitor for use in in vivo studies because it has a specific inhibition, it is permeable in cells and tissues and has low toxicity.

Uwaga dotycząca przygotowania

Working concentration: 0.5 to 1.0 μg/ml (1.4 to 28.0 μM)
Working solution: Soluble to 20 mg/ml (stock solution) in a 1:1 (v/v) mixture of ethanol and water (vortexing or slight warming in water bath (40 °C) may facilitate dissolution). Also soluble in a neutral water/methanol solution, in water, methanol, acetic acid, pyridine and DMSO. Sparingly soluble in ethanol and propanol. Insoluble in acetone, chloroform, ethyl ether and benzene.
Storage conditions (working solution): -15 to -25 °C
Solutions are stable for one month when stored in aliquots at -15 to -25 °C.

E-64 is soluble in water. A 20 mg/ml solution can be prepared in water (heat may be needed). A suggested stock solution is a 1 mM aqueous solution. The effective concentration for use as a protease inhibitor is 1 to10 μM. Aqueous stock solutions are stable for months at -20 °C. Diluted solutions are stable for days at neutral pH. E-64 is stable from pH 2-10, but is unstable in ammonia or in HCl. E-64 is also soluble in DMSO, a 10 mM solution can be prepared in dry DMSO and stored at -20 °C. Subsequent dilutions were in culture medium. Solutions for injection were prepared by dissolving E-64 in 0.9% sodium chloride or in a minimum amount of saturated sodium bicarbonate followed by dilution with 0.9% sodium chloride (after adjusting the pH to 7.0 with acetic acid).

Rekonstytucja

Soluble to 20 mg/ml in a 1:1 (v/v) mixture of ethanol and water. Solutions are stable for one month when stored in aliquots at -15 to -25 °C.

Inne uwagi

For life science research only. Not for use in diagnostic procedures.

Safety Information

• piktogramy: GHS08
• Hasło ostrzegawcze: Warning
• Zwroty wskazujące rodzaj zagrożenia: H371
• Zwroty wskazujące środki ostrożności: P260 - P264 - P270 - P308 + P311 - P405 - P501
• Klasyfikacja zagrożeń: STOT SE 2
• Kod klasy składowania: 11 - Combustible Solids
• WGK: WGK 2
• Temperatura zapłonu °F: does not flash
• Temperatura zapłonu °C: does not flash