GSK2807 TFA (GSK-2807 TFA), the trifluoroacetic acid salt form of GSK2807, is a selective and SAM-competitive SMYD3 inhibitor with potential antitumor activity. It inhibits SMYD3 with a Ki of 14 nM. SMYD3 is a lysine methyltransferase overexpressed in colorectal, breast, prostate, and hepatocellular tumors, and has been implicated as an oncogene in human malignancies. Methylation of MEKK2 by SMYD3 is important for regulation of the MEK/ERK pathway, suggesting the possibility of selectively targeting SMYD3 in RAS-driven cancers. Structural and kinetic characterization of SMYD3 was undertaken leading to a co-crystal structure of SMYD3 with a MEKK2-peptide substrate bound, and the observation that SMYD3 follows a partially processive mechanism. These insights allowed for the design of GSK2807, a potent and selective, SAM-competitive inhibitor of SMYD3 (Ki = 14 nM). A high-resolution crystal structure reveals that GSK2807 bridges the gap between the SAM-binding pocket and the substrate lysine tunnel of SMYD3. Taken together, the data demonstrate that small-molecule inhibitors of SMYD3 can be designed to prevent methylation of MEKK2 and these could have potential use as anticancer therapeutics.

Physicochemical Properties

Molecular Formula	C21H33F3N8O7
Molecular Weight	566.531334638596
Exact Mass	566.242
Elemental Analysis	C, 44.52; H, 5.87; F, 10.06; N, 19.78; O, 19.77
CAS #	2245255-66-5
Related CAS #	2245255-66-5 (TFA);2245255-65-4;
PubChem CID	134814505
Appearance	White to yellow solid powder
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	17
Rotatable Bond Count	11
Heavy Atom Count	39
Complexity	700
Defined Atom Stereocenter Count	5
SMILES	FC(C(=O)O)(F)F.O1[C@H]([C@@H]([C@@H]([C@H]1CN(CC[C@@H](C(=O)O)N)CCCN(C)C)O)O)N1C=NC2C(N)=NC=NC1=2
InChi Key	FZPZIBBKUWOBTJ-CPQMNCBCSA-N
InChi Code	InChI=1S/C19H32N8O5.C2HF3O2/c1-25(2)5-3-6-26(7-4-11(20)19(30)31)8-12-14(28)15(29)18(32-12)27-10-24-13-16(21)22-9-23-17(13)27;3-2(4,5)1(6)7/h9-12,14-15,18,28-29H,3-8,20H2,1-2H3,(H,30,31)(H2,21,22,23);(H,6,7)/t11-,12+,14+,15+,18+;/m0./s1
Chemical Name	(2S)-2-amino-4-[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl-[3-(dimethylamino)propyl]amino]butanoic acid;2,2,2-trifluoroacetic acid
Synonyms	GSK2807 TFA; GSK2807 trifluoroacetic acid; GSK2807; GSK-2807; GSK 2807
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, 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

ln Vitro	GSK2807 fills the space between SMYD3's substrate lysine tunnel and SAM binding pocket, according to the high-resolution crystal structure. Compared to the similarly related enzyme SMYD2, GSK2807 exhibits a 24 times greater selectivity for SMYD3 (Ki 14±6 nM vs. 345±36 nM, respectively) [1].
References	[1]. Structure-Based Design of a Novel SMYD3 Inhibitor that Bridges the SAM-and MEKK2-Binding Pockets. Structure. 2016 May 3;24(5):774-781. [2]. Inhibitors of Protein Methyltransferases and Demethylases. Chem Rev. 2018 Feb 14;118(3):989-1068.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~176.51 mM) H2O : ≥ 50 mg/mL (~88.26 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.67 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 20.8 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: ≥ 2.08 mg/mL (3.67 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 20.8 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: ≥ 2.08 mg/mL (3.67 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 100 mg/mL (176.51 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7651 mL	8.8257 mL	17.6513 mL
	5 mM	0.3530 mL	1.7651 mL	3.5303 mL
	10 mM	0.1765 mL	0.8826 mL	1.7651 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.