Z-DEVD-FMK, a tetrapeptide compound, is a novel, potent, specific, cell-permeable and irreversible caspase-3 inhibitor with IC50 of 18 μM. Compared to caspase-6, caspase-7, and caspase-10, it is relatively more selective for caspase-3.
Physicochemical Properties
| Molecular Formula | C30H41FN4O12 | |
| Molecular Weight | 668.66 | |
| Exact Mass | 668.27 | |
| Elemental Analysis | C, 53.89; H, 6.18; F, 2.84; N, 8.38; O, 28.71 | |
| CAS # | 210344-95-9 | |
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| PubChem CID | 16760394 | |
| Appearance | White to off-white solid powder | |
| Density | 1.3±0.1 g/cm3 | |
| Boiling Point | 914.2±65.0 °C at 760 mmHg | |
| Flash Point | 506.7±34.3 °C | |
| Vapour Pressure | 0.0±0.3 mmHg at 25°C | |
| Index of Refraction | 1.512 | |
| LogP | 4.2 | |
| Hydrogen Bond Donor Count | 4 | |
| Hydrogen Bond Acceptor Count | 13 | |
| Rotatable Bond Count | 23 | |
| Heavy Atom Count | 47 | |
| Complexity | 1110 | |
| Defined Atom Stereocenter Count | 4 | |
| SMILES | O=C(N[C@@H](C(C)C)C(N[C@H](C(CF)=O)CC(OC)=O)=O)[C@H](CCC(OC)=O)NC([C@H](CC(OC)=O)NC(OCC1=CC=CC=C1)=O)=O |
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| InChi Key | GBJVAVGBSGRRKN-JYEBCORGSA-N | |
| InChi Code | InChI=1S/C30H41FN4O12/c1-17(2)26(29(42)33-20(22(36)15-31)13-24(38)45-4)35-27(40)19(11-12-23(37)44-3)32-28(41)21(14-25(39)46-5)34-30(43)47-16-18-9-7-6-8-10-18/h6-10,17,19-21,26H,11-16H2,1-5H3,(H,32,41)(H,33,42)(H,34,43)(H,35,40)/t19-,20-,21-,26-/m0/s1 | |
| Chemical Name | methyl (4S)-5-[[(2S)-1-[[(3S)-5-fluoro-1-methoxy-1,4-dioxopentan-3-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-[[(2S)-4-methoxy-4-oxo-2-(phenylmethoxycarbonylamino)butanoyl]amino]-5-oxopentanoate | |
| Synonyms | ZDEVDfmk; Z-DEVD-fmk; Z DEVD fmk | |
| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | Caspase-3 (IC50 = 18 μM) | ||
| ln Vitro | Z-DEVD-FMK (1–200 μM) inhibits D4-GDI cleavage and apoptosis in a concentration-dependent manner. [1] Z-DEVD-FMK significantly inhibits the activation of caspase 3 and lessens ceramide-induced cardiomyocyte death. [3] In cultured brain microvessel endothelial cells, Z-DEVD-FMK (100 μM) inhibits OxyHb's effects on DNA ladders, caspase-2 and -3 activity, cell detachment, and PARP cleavage. [4] Caspase-3 activity increases brought on by MPP+ are prevented by Z-DEVD-FMK (100 μM). With an IC50 of 18 μM, Z-DEVD-FMK inhibits 6-OHDA-induced apoptotic cell death in a dose-dependent manner. [5] | ||
| ln Vivo | Z-DEVD-FMK significantly improves neurological recovery both before and after injury by significantly reducing post-traumatic apoptosis. [2] | ||
| Enzyme Assay | Utilizing fluorescent-based substrate, the activities of caspase-3 and caspase-9 are measured. Following treatment, the cells are resuspended in 10 mM digitonin-containing lysis buffer (50 mM Tris HCl, 1 mM EDTA, and 10 mM EGTA) for 20 min at 37°C. Ac-DEVD-AFC for caspase-3 or Ac-LEHD-AFC for caspase-9 are applied to supernatants for 1 hour at 37 degrees Celsius. Fluorescence is then measured using a Gemini XS fluorescence plate reader at excitation at 400 nm and emission at 505 nm. | ||
| Cell Assay | Using the widely used MTT assay (3-(4,5-dimethylthiazol-3-yl)-2,5-diphenyl tetrazolium bromide), cell death in N27 cells is assessed after incubation with 100 μM 6-OHDA for 24 h or 300 μM MPP+ for 36 h in the presence or absence of 50 μM Z-DEVD-FMK. The cells are then treated and incubated for 3 hours at 37 degrees Celsius in serum-free medium containing 0.25 mg/ml MTT. Using a SpectraMax microplate reader and a reference wavelength of 630 nm, the formation of formazan from tetrazolium is measured at 570 nm. | ||
| Animal Protocol |
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| References |
[1]. Eur J Immunol . 1998 Jan;28(1):296-304. [2]. J Neurosci . 1997 Oct 1;17(19):7415-24. [3]. J Card Fail . 2000 Sep;6(3):243-9. [4]. Stroke . 2001 Feb;32(2):561-6. [5]. Free Radic Biol Med . 2006 Nov 15;41(10):1578-89. |
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| Additional Infomation | Z-DEVD-FMK is a tetrapeptide consisting of Fmoc-L-aspartic acid 4-methyl ester, methyl L-alpha-glutamic acid 5-methyl ester, L-valine and the fluoromethyl ketone derived from the 1-carboxy group of L-aspartic acid 4-methyl ester coupled in sequence. It is a specific, irreversible caspase-3 inhibitor that also shows potent inhibition of caspase-6, caspase-7, caspase-8, and caspase-10. It has a role as an apoptosis inhibitor, an EC 3.4.22.56 (caspase-3) inhibitor and a neuroprotective agent. |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (1.50 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (1.50 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 2% DMSO+30% PEG 400+5% Tween 80+ddH2O: 1mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4955 mL | 7.4776 mL | 14.9553 mL | |
| 5 mM | 0.2991 mL | 1.4955 mL | 2.9911 mL | |
| 10 mM | 0.1496 mL | 0.7478 mL | 1.4955 mL |