AES-350 is a potent orally bioactive HDAC6 inhibitor (antagonist) with IC50 and Ki of 0.0244 μM and 0.035 μM, respectively. The IC50s of AES-350 for HDAC-3, -8 and -11 are 0.187 μM and 0.245 μM respectively. AES-350 induces AML cell apoptosis by inhibiting HDAC and may be utilized in acute myeloid leukemia (AML) research.

Physicochemical Properties

Molecular Formula	C18H20N2O3
Molecular Weight	312.363
Exact Mass	312.147
Elemental Analysis	C, 69.21; H, 6.45; N, 8.97; O, 15.37
CAS #	847249-57-4
Related CAS #	847249-57-4
PubChem CID	11688197
Appearance	Off-white to light yellow solid powder
LogP	4.13
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	4
Heavy Atom Count	23
Complexity	415
Defined Atom Stereocenter Count	0
SMILES	CC(C1=CC=C(C(NC2=CC=C(C(NO)=O)C=C2)=O)C=C1)(C)C
InChi Key	FMOQHLZNJFXULZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H20N2O3/c1-18(2,3)14-8-4-12(5-9-14)16(21)19-15-10-6-13(7-11-15)17(22)20-23/h4-11,23H,1-3H3,(H,19,21)(H,20,22)
Chemical Name	4-tert-butyl-N-[4-(hydroxycarbamoyl)phenyl]benzamide
Synonyms	AES 350; AES-350; AES350
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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	HDAC6 (IC50 = 24.4 nM); HDAC3 (IC50 = 187 nM); HDAC11 (IC50 = 245 nM)
ln Vitro	On the other hand, AES-350 exhibits submicromolar activity (IC50=0.58±0.13 μM) in comparison to vorinostat (IC50=0.31±0.061 μM) against MV4-11 cells. AES-350 exhibits greater ligand efficiency and a broad therapeutic index (IC50>30 μM in MRC-9 cells that are not cancerous). In AML-3 (acute myeloid leukemia) cells, AES-350 has also been demonstrated to be effective (IC50=0.73 ± 0.12 μM)[1]. AES-350 (0.25–4 μM; 18 hours) causes a dose-dependent apoptosis in MV4-11 cells. At a concentration of 0.25 μM–4 μM, the late apoptosis ratios are 8.74%, 11.7%, 16.08%, 30.97%, and 38.48%, respectively[1]. HeLa cervical cancer cell lysates are used for an ELISA; these cells exhibit high levels of HDAC6 expression and are AES-350 sensitive. AES-350 (0.1-10 μM) causes a dose-dependent increase in acetylated α-tubulin (Ac-α-tubulin), a substrate of HDAC. ELISA assays similarly showed a dose-dependent increase in HDAC6 inhibition (IC50=0.58±0.13 μM)[1].
ln Vivo	AES-350 (oral gavage; 20 mg/kg; single dose) shows comparatively good pharmacokinetic (PK) characteristics in CD-1 mice. The single dose oral bioavailability (F%) of 51 is 19.8%. In comparison, the reported F% for SAHA in mice is significantly lower (8%)[1].
Cell Assay	Cell Line: MV4-11 cells Concentration: 0.25 μM; 0.5 μM; 1.00 μM; 2.00 μM; 4.00 μM Incubation Time: 18 hours Result: Revealed a clear dosedependent increase in the percentage of cells entering late-stage apoptosis, similar to SAHA.
References	[1]. Class I/IIb-Selective HDAC Inhibitor Exhibits Oral Bioavailability and Therapeutic Efficacy in Acute Myeloid Leukemia.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~320.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.00 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 25.0 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: ≥ 2.5 mg/mL (8.00 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 25.0 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: ≥ 2.5 mg/mL (8.00 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 25.0 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	3.2014 mL	16.0072 mL	32.0143 mL
	5 mM	0.6403 mL	3.2014 mL	6.4029 mL
	10 mM	0.3201 mL	1.6007 mL	3.2014 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.