ZYZ-488 is a competitive apoptotic protease activating factor-1 (Apaf-1) inhibitor that can suppress activation of the binding proteins procaspase-9 and procaspase-3.

Physicochemical Properties

Molecular Formula	C20H29N3O11
Molecular Weight	487.4578
Exact Mass	487.18
CAS #	1470302-79-4
PubChem CID	138454738
Appearance	White to off-white solid powder
LogP	-1
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	12
Rotatable Bond Count	12
Heavy Atom Count	34
Complexity	691
Defined Atom Stereocenter Count	5
SMILES	O1[C@]([H])([C@@]([H])([C@]([H])([C@@]([H])([C@@]1([H])C(=O)O[H])O[H])O[H])O[H])OC1C(=C([H])C(C(=O)OC([H])([H])C([H])([H])C([H])([H])C([H])([H])/N=C(\N([H])[H])/N([H])[H])=C([H])C=1OC([H])([H])[H])OC([H])([H])[H]
InChi Key	PVTAIJDQJJZCJP-WXZWTWHISA-N
InChi Code	InChI=1S/C20H29N3O11/c1-30-10-7-9(18(29)32-6-4-3-5-23-20(21)22)8-11(31-2)15(10)33-19-14(26)12(24)13(25)16(34-19)17(27)28/h7-8,12-14,16,19,24-26H,3-6H2,1-2H3,(H,27,28)(H4,21,22,23)/t12-,13-,14+,16-,19+/m0/s1
Chemical Name	(2S,3S,4S,5R,6S)-6-[4-[4-(diaminomethylideneamino)butoxycarbonyl]-2,6-dimethoxyphenoxy]-3,4,5-trihydroxyoxane-2-carboxylic acid
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.
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	Apoptotic protease activating factor-1[1]
ln Vitro	In hypoxic H9c2 cells, ZYZ-488 reduces apoptosis. ZYZ-488 suppresses procaspase-9 and procaspase-3 activation mediated by Apaf-1 in response to hypoxia[1]. ZYZ-488 suppresses the activation of the downstream caspases cascade, prevents cardiomyocyte death, and reduces the release of cardiac enzymes[2].
ln Vivo	ZYZ-488 (intramuscular; 33.9–67.8 mg/kg; 3 days) prevented myocardial apoptosis[2].
Animal Protocol	Animal/Disease Models: Male C57BL/6 mice (8-10 weeks old)[2] Doses: myocardial infarction + ZYZ-488 (33.9 mg/kg); myocardial infraction + ZYZ-488 (67.8 mg/kg); Route of Administration: Intramuscularly; for 3 days Experimental Results: Inhibited Apaf- 1-mediated activation of procaspase-9 and procaspase-3.
References	[1]. The discovery of a novel inhibitor of apoptotic protease activating factor-1 (Apaf-1) for ischemic heart: synthesis, activity and target identification. Sci Rep. 2016 Jul 22;6:29820.

Solubility Data

Solubility (In Vitro)

DMSO : 200 mg/mL (410.29 mM) H2O : ≥ 100 mg/mL (205.15 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 5 mg/mL (10.26 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 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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: ≥ 5 mg/mL (10.26 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 50.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 5 mg/mL (10.26 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 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.0515 mL	10.2573 mL	20.5145 mL
	5 mM	0.4103 mL	2.0515 mL	4.1029 mL
	10 mM	0.2051 mL	1.0257 mL	2.0515 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.