5.31661

SMYD2 Inhibitor, AZ505

• Synonym(s): SMYD2 Inhibitor, AZ505, N-Cyclohexyl-3-(3,4-dichlorophenethylamino)-N-(2-(2-(5-hydroxy-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)ethylamino)ethyl)propanamide, HMTase Inhibitor XV
• Empirical Formula (Hill Notation): C₂₉H₃₈Cl₂N₄O₄
• CAS Number: 1035227-43-0
• Molecular Weight: 577.54
• UNSPSC Code: 12352200

Properties

• Assay: ≥95% (HPLC)
• Quality Level: 100
• form: solid
• manufacturer/tradename: Calbiochem^®
• storage condition: OK to freeze; protect from light
• color: off-white
• solubility: DMSO: 50 mg/mL
• storage temp.: 2-8°C
• InChI: 1S/C29H38Cl2N4O4/c30-23-8-6-20(18-24(23)31)10-13-32-15-12-27(38)35(22-4-2-1-3-5-22)17-16-33-14-11-21-7-9-25(36)28-29(21)39-19-26(37)34-28/h6-9,18,22,32-33,36H,1-5,10-17,19H2,(H,34,37)
• InChI key: LIBVHXXKHSODII-UHFFFAOYSA-N

Description

General description

A cell-permeable benzooxazinyl-ethylaminoethyl-propanamide compound that acts as a potent and reversible inhibitor of SMYD2 (IC₅₀ = 120 nM, K_i = 300 nM, K_d = 500 nM). Shown to bind to the peptide-binding groove of the enzyme. This binding is dependent on the presence of S-adenosylmethionine (SAM). The inhibition appears to be competitive with respect to substrate and uncompetitive with respect to SAM. Displays ~700-fold greater selectivity over SMYD3, DOT1L, EZH2, GLP, G9a and SET7/9 protein lysine methyl transferases (IC₅₀ >83.3 µM). Inhibits SMYD2-mediated p53-K370 methylation in U2OS cells (at ~10 µM), but does not affect methylation in cells transfected with a Y240F mutant.

Please note that the molecular weight for this compound is batch-specific due to variable water content.

A cell-permeable benzooxazinyl-ethylaminoethyl-propanamide compound that inhibits SMYD2-mediated p53 peptide (aa 361-380) K370me monomethylation (IC₅₀ = 120 nM; [SAM] = [p53 peptide] = 1 µM; pH 7.4 for 90 min) in a substrate-competitive, but cofactor S-adenosylmethionine-(AdoMet/SAM) uncompetitive manner (K_i = 0.3 µM) by targeting SAM-bound SMYD2 at the substrate-binding site (K_d = 0.5 µM) via simultaneous interactions with SAM donor methyl group as well as residues from the core SET, I-SET, and post-SET domains, while exhibiting little potency toward DOT1L, EZH2, G9A, GLP, SET7/9, or SMYD3 (IC₅₀ >83.3 µM). Shown to effectively abolish cellular p53 K370me in U2OS wt SMYD2 transfectants to the same level seen in U2OS transfected with inactive SMYD2 Y240F mutant (10 µM for 24 h).

AZ505 is a cell-permeable, potent, reversible inhibitor of SMYD2 (IC₅₀ = 120 nM, K_i = 300 nM) that binds to the peptide-binding groove of the enzyme.

Biochem/physiol Actions

Cell permeable: yes

Primary Target
SMYD2

Warning

Toxicity: Standard Handling (A)

Physical form

Supplied as a di-trifluoroacetate salt.

Reconstitution

Following reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.

Use only fresh DMSO for reconstitution.

Other Notes

Ferguson, A.D., et al. 2011. Structure19, 1262.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable