E 64d (Synonyms: Aloxistatin, E64c ethyl ester, EP 453, EST, Loxistatin, NSC 694281) |
Catalog No.GC13787
One-Click Copy Product Info
|
E 64d is a cell-permeable, irreversible, broad-spectrum cysteine protease inhibitor with platelet aggregation inhibitory activity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 88321-09-9
Sample solution is provided at 25 µL, 10mM.
E 64d is a cell-permeable, irreversible, broad-spectrum cysteine protease inhibitor with platelet aggregation inhibitory activity[1]. E 64d is a non-selective covalent tissue protease inhibitor containing an epoxide ring as a warhead that can be nucleophilically attacked by Cys-SH and form a covalent bond at the catalytic site[2]. E 64d inhibits the accumulation of protease-resistant prion protein (PrP-res) in ScNB cells with an IC50 of 0.5±0.11μM[3].
In vitro, pretreatment of SH-SY5Y cells with E 64d (3µM) for 30min improved the amyloid beta-induced decrease in sAPPα secretion and reversed the ceramide-induced downregulation of protein kinase C[4]. Treatment of HT22 cells with E 64d (25μM) for 24h alleviated glutamate-induced apoptosis, reversed the decrease in mitochondrial membrane potential, and reduced oxidative stress[5].
In vivo, E 64d (9mg/kg/day) was intraperitoneally injected into arthritic mice for 10 days, which significantly reduced the severity of clinical arthritis and histopathology, and decreased the mRNA levels of cytokines IL-6 and IL-1β[6]. E 64d (1mg/kg) was intravenously injected into rats with spinal cord injury (SCI) for 7 days, which inhibited the inflammatory response caused by SCI, reduced the expression of calretinin 1 in spinal cord tissue, reduced neuronal apoptosis, and inhibited the activity of cyclooxygenase 2[7].
References:
[1] Chen Y, Li Q, Li Q, et al. p62/SQSTM1, a central but unexploited target: advances in its physiological/pathogenic functions and small molecular modulators[J]. Journal of medicinal chemistry, 2020, 63(18): 10135-10157.
[2] Cannalire R, Stefanelli I, Cerchia C, et al. SARS-CoV-2 entry inhibitors: small molecules and peptides targeting virus or host cells[J]. International Journal of Molecular Sciences, 2020, 21(16): 5707.
[3] Doh-Ura K, Iwaki T, Caughey B. Lysosomotropic agents and cysteine protease inhibitors inhibit scrapie-associated prion protein accumulation[J]. Journal of virology, 2000, 74(10): 4894-4897.
[4] 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[J]. Biochemical and Biophysical Research Communications, 2013, 441(1): 256-261.
[5] Xie R J, Li T X, Qiao X Y, et al. The Protective Role of E‐64d in Hippocampal Excitotoxic Neuronal Injury Induced by Glutamate in HT22 Hippocampal Neuronal Cells[J]. Neural Plasticity, 2021, 2021(1): 7174287.
[6] Yoshifuji H, Umehara H, Maruyama H, et al. Amelioration of experimental arthritis by a calpain-inhibitory compound: regulation of cytokine production by E-64-d in vivo and in vitro[J]. International immunology, 2005, 17(10): 1327-1336.
[7] Zhang Z, Huang Z, Dai H, et al. Therapeutic Efficacy of E‐64‐d, a Selective Calpain Inhibitor, in Experimental Acute Spinal Cord Injury[J]. BioMed Research International, 2015, 2015(1): 134242.
| Cell experiment [1]: | |
Cell lines | SH-SY5Y cells |
Preparation Method | SH-SY5Y cells were pretreated with or without E 64d (3µM) for 30min and incubated with Aβ (0.001 or 0.01µM) for 5.5h, followed by incubation with methacholine (1mM) for 2h. Secreted sAPPα was analyzed by Western blotting using 6E10 antibody. |
Reaction Conditions | 3µM; 30min |
Applications | E 64d restores the Aβ-induced reduction of sAPPα secretion in SH-SY5Y cells. |
| Animal experiment [2]: | |
Animal models | BALB/c mice |
Preparation Method | Arthritis was induced in 7-week-old female BALB/c mice with a mixture of anti -type II CL mAbs and LPS. 2 mg per body of mAb mixture was injected i.p. into each mouse, and 48h later, 50µg per body of LPS was injected i.p. in order to enhance the arthritis. A high dose (9mg/kg/day) or low dose (3mg/kg/day) of E 64d, recombinant cal pastatin (rCS) (9mg/kg/day) or control diluent was administered i.p. to the disease-affected mice for 10 days from the day before the injection of mAbs. Excise the right hind paw from each body and macroscopically assess the severity of arthritis in each paw. Evaluate histopathological severity based on findings of cellular infiltration, pannus formation, and bone erosion. |
Dosage form | 3、9mg/kg/day for 10 days; i.p. |
Applications | The clinical arthritis score of the high-dose E 64d group was significantly lower than that of the untreated group, while there was no improvement in the low-dose E 64d group and the rCS-treated group. The cell infiltration histopathological score and total histopathological arthritis score of the high-dose E 64d group were significantly lower than those of the untreated group. |
References: [1]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[J]. Biochemical and Biophysical Research Communications, 2013, 441(1): 256-261. [2]Yoshifuji H, Umehara H, Maruyama H, et al. Amelioration of experimental arthritis by a calpain-inhibitory compound: regulation of cytokine production by E-64-d in vivo and in vitro[J]. International immunology, 2005, 17(10): 1327-1336. | |
| Cas No. | 88321-09-9 | SDF | |
| Synonyms | Aloxistatin, E64c ethyl ester, EP 453, EST, Loxistatin, NSC 694281 | ||
| Chemical Name | ethyl (2S,3S)-3-[[(2S)-4-methyl-1-(3-methylbutylamino)-1-oxopentan-2-yl]carbamoyl]oxirane-2-carboxylate | ||
| Canonical SMILES | CCOC(=O)C1C(O1)C(=O)NC(CC(C)C)C(=O)NCCC(C)C | ||
| Formula | C17H30N2O5 | M.Wt | 342.43 |
| Solubility | ≥ 17.1215mg/mL in DMSO | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.9203 mL | 14.6015 mL | 29.203 mL |
| 5 mM | 584.1 μL | 2.9203 mL | 5.8406 mL |
| 10 mM | 292 μL | 1.4602 mL | 2.9203 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)