MAK-683 (MAK683; extracted from patent US20160176882 A1, compound example 2) is a novel and potent embryonic ectoderm development protein (EED) inhibitor and allosteric inhibitor of polycomb repressive complex 2 (PRC2) with potential anticancer activity. It inhibits EED and PRC2 with IC50s of 59, 89, 26 nM in EED Alphascreen binding, LC-MS and ELISA assay.

Physicochemical Properties

Molecular Formula	C20H17FN6O
Molecular Weight	376.386986494064
Exact Mass	376.144
CAS #	1951408-58-4
Related CAS #	MAK683 hydrochloride;2170606-94-5
PubChem CID	121412508
Appearance	White to off-white solid powder
LogP	3.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	541
Defined Atom Stereocenter Count	0
SMILES	FC1C=CC2=C(C=1CNC1=NC=C(C3=CC=CN=C3C)C3=NN=CN13)CCO2
InChi Key	XLIBABIFOBYHSV-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H17FN6O/c1-12-13(3-2-7-22-12)16-10-24-20(27-11-25-26-19(16)27)23-9-15-14-6-8-28-18(14)5-4-17(15)21/h2-5,7,10-11H,6,8-9H2,1H3,(H,23,24)
Chemical Name	N-[(5-fluoro-2,3-dihydro-1-benzofuran-4-yl)methyl]-8-(2-methylpyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine
Synonyms	MAK683 MAK 683 MAK-683 EED inhibitor-1 EED inhibitor 1 EED inhibitor1
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
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	After 14 days of therapy, MAK683 has an IC50 of 30 nM against B-cell lymphoma cells KARPAS422 that exhibits anti-proliferative action [1].
References	[1]. TRIAZOLOPYRIMIDINE COMPOUNDS AND USES THEREOF. US20160176882 A1. [2]. Binding Modes of Small-Molecule Inhibitors to the EED Pocket of PRC2. Chemphyschem. 2020 Feb 4;21(3):263-271.
Additional Infomation	EED Inhibitor MAK683 is an inhibitor of embryonic ectoderm development protein (EED) and allosteric inhibitor of polycomb repressive complex 2 (PRC2), with potential antineoplastic activity. Upon administration, MAK683 selectively binds to the domain of EED that interacts with trimethylated lysine 27 on histone 3 (H3K27me3), which leads to a conformational change in the EED H3K27me3-binding pocket and prevents the interaction of EED with the histone methyltransferase enhancer zeste homolog 2 (EZH2). Disruption of the EED-EZH2 protein-protein interaction (PPI) results in a loss of H3K27me3-stimulated PRC2 activity and prevents H3K27 trimethylation. This decrease in histone methylation alters gene expression patterns associated with cancer pathways and results in decreased tumor cell proliferation in EZH2-mutated and PRC2-dependent cancer cells. PRC2, a histone H3 lysine 27 methyltransferase and multi-protein complex comprised of EZH2, EED and suppressor of zeste 12 (SUZ12), plays a key role in gene regulation, especially during embryonic development. EZH2, the catalytic subunit of PRC2, is overexpressed or mutated in a variety of cancer cells. EED is essential for the histone methyltransferase activity of PRC2 because EED directly binds to H3K27me3.

Solubility Data

Solubility (In Vitro)

DMSO : ~40 mg/mL (~106.27 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.17 mg/mL (5.77 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 21.7 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: ≥ 2.17 mg/mL (5.77 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 21.7 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	2.6568 mL	13.2841 mL	26.5682 mL
	5 mM	0.5314 mL	2.6568 mL	5.3136 mL
	10 mM	0.2657 mL	1.3284 mL	2.6568 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.