Ac-DEVD-CHO is a potent and specific aldehyde inhibitor of Group II caspases with Ki values of 0.2 nM and 0.3 nM for for caspase-3 and caspase-7, respectively. Weak caspase-2 inhibition. This aldehyde only moderately inhibits caspase-2 (Ki = 1.7 μM), which only cleaves the tetrapeptide substrate. Ac-DEVD-CHO has a broad inhibitory effect on group III caspases, with Ki values ranging from 1 to 300 nM. Even when given after the onset of ischemia, Ac-DEVD-CHO'scaspase-3inhibition significantly improves the stunned myocardium's post-ischemic contractile recovery in the isolated working-heart rat model. Ac-DEVD-CHO appears to have protection mechanisms that are separate from apoptosis. Ac-DEVD-CHO did not prevent troponin I cleavage.

Physicochemical Properties

Molecular Formula	C20H30N4O11
Molecular Weight	502.47
Exact Mass	502.191
Elemental Analysis	C, 47.81; H, 6.02; N, 11.15; O, 35.02
CAS #	169332-60-9
Related CAS #	169332-60-9
PubChem CID	644345
Sequence	N-Acetyl-Asp-Glu-Val-Asp-al
SequenceShortening	Ac-DEVD-al
Appearance	White to off-white solid powder
Density	1.374g/cm3
Boiling Point	1021.1ºC at 760mmHg
Flash Point	571.3ºC
Vapour Pressure	0mmHg at 25°C
Index of Refraction	1.535
LogP	-2.6
Hydrogen Bond Donor Count	7
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	16
Heavy Atom Count	35
Complexity	843
Defined Atom Stereocenter Count	4
SMILES	O=C([C@]([H])(C([H])([H])C([H])([H])C(=O)O[H])N([H])C([C@]([H])(C([H])([H])C(=O)O[H])N([H])C(C([H])([H])[H])=O)=O)N([H])[C@]([H])(C(N([H])[C@]([H])(C([H])=O)C([H])([H])C(=O)O[H])=O)C([H])(C([H])([H])[H])C([H])([H])[H]
InChi Key	UMBVAPCONCILTL-MRHIQRDNSA-N
InChi Code	InChI=1S/C20H30N4O11/c1-9(2)17(20(35)22-11(8-25)6-15(29)30)24-18(33)12(4-5-14(27)28)23-19(34)13(7-16(31)32)21-10(3)26/h8-9,11-13,17H,4-7H2,1-3H3,(H,21,26)(H,22,35)(H,23,34)(H,24,33)(H,27,28)(H,29,30)(H,31,32)/t11-,12-,13-,17-/m0/s1
Chemical Name	(4S)-4-[[(2S)-2-acetamido-3-carboxypropanoyl]amino]-5-[[(2S)-1-[[(2S)-1-carboxy-3-oxopropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-5-oxopentanoic acid
Synonyms	Caspase-3 Inhibitor I; N-Ac-Asp-Glu-Val-Asp-CHO; Ac-DEVD CHO; Ac DEVD-CHO; Ac DEVD CHO
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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 (Ki = 0.23 nM); Caspase-8 (Ki = 0.92 nM); Caspase-7 (Ki = 1.6 nM); Caspase-10 (Ki = 12 nM); Caspase-1 (Ki = 18 nM); Caspase-6 (Ki = 31 nM); Caspase-9 (Ki = 60 nM); Caspase-4 (Ki = 132 nM); Caspase-5 (Ki = 205 nM); Caspase-2 (Ki = 1710 nM)
ln Vitro	Ac-DEVD-CHO is a potent inhibitor of caspase-3 (Ki = 230 pM). In contrast, this aldehyde only slightly inhibits caspase-2 (Ki = 1.7 μM) and exhibits poor cleavage of the tetrapeptide substrate. With Ki values ranging from 1 to 300 nM, Ac-DEVD-CHO significantly inhibits Group III caspases[1]. Even when administered after the onset of ischemia, Ac-DEVD-CHO'scaspase-3inhibition significantly improves post-ischemic contractile recovery of stunned myocardium in the isolated working-heart rat model. Ac-DEVD-CHO'sprotectivemechanism(s) seem to operate independently of apoptosis. Ac-DEVD-CHO[2] did not inhibit troponin I cleavage.
ln Vivo	Receiving Ac-DEVD-CHO at the time of MI causes a 61% decrease in the expression of activated caspase-3 in cardiomyocytes (p<0.05) and an 84% decrease in cardiomyocyte apoptosis in young animals. Caspase inhibition, however, had no impact on cardiomyocyte apoptosis or activated caspase-3 expression in the aging mice[4]. Ac-DEVD-CHO inhibited and/or postponed the development of photoreceptor cell damage in rats, as well as slows the disease's progression in rd gene-carrying mice, which typically experience retinal degeneration in their early years of life[2].
Enzyme Assay	Studies with peptide-based and macromolecular inhibitors of the caspase family of cysteine proteases have helped to define a central role for these enzymes in inflammation and mammalian apoptosis. A clear interpretation of these studies has been compromised by an incomplete understanding of the selectivity of these molecules. Here we describe the selectivity of several peptide-based inhibitors and the coxpox serpin CrmA against 10 human caspases. The peptide aldehydes that were examined (Ac-WEHD-CHO, Ac-DEVD-CHO, Ac-YVAD-CHO, t-butoxycarbonyl-IETD-CHO, and t-butoxycarbonyl-AEVD-CHO) included several that contain the optimal tetrapeptide recognition motif for various caspases. These aldehydes display a wide range of selectivities and potencies against these enzymes, with dissociation constants ranging from 75 pM to >10 microM. The halomethyl ketone benzyloxycarbonyl-VAD fluoromethyl ketone is a broad specificity irreversible caspase inhibitor, with second-order inactivation rates that range from 2.9 x 10(2) M-1 s-1 for caspase-2 to 2.8 x 10(5) M-1 s-1 for caspase-1. The results obtained with peptide-based inhibitors are in accord with those predicted from the substrate specificity studies described earlier. The cowpox serpin CrmA is a potent (Ki < 20 nM) and selective inhibitor of Group I caspases (caspase-1, -4, and -5) and most Group III caspases (caspase-8, -9, and -10), suggesting that this virus facilitates infection through inhibition of both apoptosis and the host inflammatory response[1].
Cell Assay	OCLs are incubated with RANKL and treated with 0.5 mM SIN for 24 hours, either with or without the particular caspase-3 inhibitor Ac-DEVD-CHO (10 μM). After the treatment, the cells are rinsed with PBS and stained for 15 min with 10 μM Hoechst 33258 dye. A fluorescent microscope is used to take pictures of the staining cells. By counting the number of cells with apoptotic nuclear condensation in each well, the differences are measured[4].
Animal Protocol	One hundred and two male mice are subjected to cecal ligation and puncture or sham operation. The animals are assigned into three equal groups (n=34) according to random number table: sham group, model group, and caspase-3 inhibitor (CI) group. Thirty minutes before CLP, Ac-DEVD-CHO (4 μg/g) is injected subcutaneously in CI group. The levels of blood urea nitrogen (BUN) and creatinine (Cr) are determined, and the concentrations of tumor necrosis factor-α (TNF-α), interleukins (IL-6 and IL-10) are measured by enzyme linked immunosorbent assay (ELISA), the renal cell apoptosis rate is determined by flow cytometry. The 4-day and 7-day survival rates of three groups of mice are observed[5]. 3 mg/kg; i.p. C57Bl6 mice.
References	[1]. J Biol Chem . 1998 Dec 4;273(49):32608-13. [2]. J Am Coll Cardiol . 2001 Dec;38(7):2063-70. [3]. Mol Cell Biol . 2006 Nov;26(21):7880-91. [4]. Cardiovasc Ther . 2013 Dec;31(6):e102-10. [5] Acta Histochem. 2003, 36(4):263-270.
Additional Infomation	Ac-Asp-Glu-Val-Asp-H is a tetrapeptide consisting of two L-aspartic acid residues, an L-glutamyl residue and an L-valine residue with an acetyl group at the N-terminal and with the C-terminal carboxy group reduced to an aldehyde. It is an inhibitor of caspase-3/7. It has a role as a protease inhibitor.

Solubility Data

Solubility (In Vitro)	Note:Please refer to page 4 in the "Instructions for use" file (upper-right section of this webpage) for how to dissolve peptides. DMSO: ~10 mM Water: ~100 mg/mL (~199.0 mM) Ethanol: N/A
Solubility (In Vivo)	Solubility in Formulation 1: 100 mg/mL (199.02 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9902 mL	9.9508 mL	19.9017 mL
	5 mM	0.3980 mL	1.9902 mL	3.9803 mL
	10 mM	0.1990 mL	0.9951 mL	1.9902 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.