(2R,3R)-3-{[(1S)-1-[(4-carbamimidamidobutyl)carbamoyl]-3-methylbutyl]carbamoyl}oxirane-2-carboxylic acid

• ChemBase ID: 132401
• Molecular Formular: C15H27N5O5
• Molecular Mass: 357.40538
• Monoisotopic Mass: 357.20121899
• SMILES: CC(C)C[C@@H](C(=O)NCCCCNC(=N)N)NC(=O)[C@H]1[C@@H](O1)C(=O)O
• Canonical SMILES: CC(C[C@@H](C(=O)NCCCCNC(=N)N)NC(=O)[C@@H]1O[C@H]1C(=O)O)C
• InChI: 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
• InChIKey: LTLYEAJONXGNFG-HBNTYKKESA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: (2R,3R)-3-{[(1S)-1-[(4-carbamimidamidobutyl)carbamoyl]-3-methylbutyl]carbamoyl}oxirane-2-carboxylic acid
• IUPAC Traditional name: (2R,3R)-3-{[(1S)-1-[(4-carbamimidamidobutyl)carbamoyl]-3-methylbutyl]carbamoyl}oxirane-2-carboxylic acid
• Synonyms: trans-Epoxysuccinyl-L-leucylamido(4-guanidino)butane; L-trans-3-Carboxyoxiran-2-carbonyl-L-leucylagmatine; N-(trans-Epoxysuccinyl)-L-leucine 4-guanidinobutylamide; E-64

Database IDs

• CAS Number: 66701-25-5
• MDL Number: MFCD00080261
• Beilstein Number: 1405664
• PubChem SID: 24278411; 24868843; 162226678
• PubChem CID: 439487

CALCULATED PROPERTIES

• Acid pKa: 3.3650656
• H Acceptors: 8
• H Donor: 6
• LogD (pH = 5.5): -2.7337623
• LogD (pH = 7.4): -2.7311778
• Log P: -2.7311957
• Molar Refractivity: 98.5676 cm^3
• Polarizability: 34.432243 Å^3
• Polar Surface Area: 169.93 Å^2
• Rotatable Bonds: 11
• Lipinski's Rule of Five: false

PROPERTIES

Physical Property

• Solubility: H2O: soluble1 mM (Stock Solutions stable for months at –20° C.)

Safety Information

• RTECS: RR0390000
• German water hazard class: 3
• Personal Protective Equipment: Eyeshields, Gloves, type N95 (US), type P1 (EN143) respirator filter
• Storage Temperature: 2-8°C

Product Information

• Purity: ≥99.0% (TLC)
• Empirical Formula (Hill Notation): C15H27N5O5

DETAILS

Sigma Aldrich - E3132

Preparation Note
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).
Application
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.1
Biochem/physiol Actions
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.

Sigma Aldrich - 45370

Preparation Note
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).
Application
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.1
Biochem/physiol Actions
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.