CeMMEC1, an N-methylisoquinolinone analog, is a novel and potent inhibitor of TAF4 with anticancer activity. Moreover, it inhibits the second bromodomain of TAF1 by blocking BRD4 (Kd = 1.8 µM; IC50 = 0.9 µM). CeMMEC1 only very weakly bound BRD4, but it exhibited strong affinity for the second bromodomain of TAF1, CREBBP, EP300, and BRD9. It doesn't attach to BRD4's first or second bromodomains. Bromodomain pharmacological inhibitors offer potential treatment advantages for a range of malignancies.

Physicochemical Properties

Molecular Formula	C19H16N2O4
Molecular Weight	336.341344833374
Exact Mass	336.11
Elemental Analysis	C, 67.85; H, 4.80; N, 8.33; O, 19.03
CAS #	440662-09-9
Related CAS #	440662-09-9
PubChem CID	24152379
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	590.1±50.0 °C at 760 mmHg
Flash Point	310.7±30.1 °C
Vapour Pressure	0.0±1.7 mmHg at 25°C
Index of Refraction	1.673
LogP	2.69
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	25
Complexity	576
Defined Atom Stereocenter Count	0
SMILES	O1CCOC2C=CC(=CC1=2)NC(C1=CN(C)C(C2C=CC=CC1=2)=O)=O
InChi Key	PEOQAZBGLOADFJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H16N2O4/c1-21-11-15(13-4-2-3-5-14(13)19(21)23)18(22)20-12-6-7-16-17(10-12)25-9-8-24-16/h2-7,10-11H,8-9H2,1H3,(H,20,22)
Chemical Name	N-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-methyl-1-oxoisoquinoline-4-carboxamide
Synonyms	Ce MMEC1; Ce-MMEC1; CeMMEC1
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

Targets	TAF1(2) ( Kd = 1.8 μM ); TAF1 ( IC50 = 0.9 μM )
ln Vitro	CeMMEC1, a BRD4 inhibitor, exhibits a high affinity for TAF1, as evidenced by its IC50 of 0.9 μM and Kd of 1.8 μM (2) additionally demonstrates a strong affinity for the BRD9, EP300, and CREBBP bromodomains. CeMMEC1, at doses of 1, 10, and 20 μM, reduces the quantity of THP1 cells in the S phase. Moreover, CeMMEC1 triggers apoptosis. When CeMMEC1 and (S)-JQ1 are combined, treatment alone significantly reduces cell viability[1].
Enzyme Assay	The EPIgeneous Binding Domain kit B is used for TAF1 binding assays. The displacement of an acetylated biotin peptide from a GST-tagged TAF1 protein using HTRF in conjunction with a streptavidin-conjugated acceptor and an Eu3+-conjugated GST antibody donor indicates binding. Using an Echo 525 Liquid Handler, compounds (CeMMEC1) are dispensed into ProxiPlate-384 Plus assay plates. 5 nM TAF1-GST, 50 nM peptide (SGRGK (ac)GGK (ac)GGAK (ac)RHRK (biotin)-acid), 6.25 nM Streptavidin-XL665, 1:200 Anti-GST-Eu3+ cryptate, and 0.1% DMSO are used in binding assays, which are carried out in a final volume of 20 μL. Utilizing a Multidrop combi, assay reagents are applied to plates and allowed to sit at room temperature for three hours. A PHERAstar microplate reader with the HTRF module and dual emission protocol is used to measure fluorescence (A = 320 nm excitation, 665 nm emission, and B = 320 nm excitation, 620 nm emission). HTRF ratio (channel A/B × 10,000) is obtained by processing raw data and is utilized in the creation of IC50 curves[1].
Cell Assay	Target genes like the MYC oncogene are transcriptionally activated by proteins of the BET family that contain a bromodomain and can detect acetylation of histone lysine. BET domain pharmacological inhibitors offer potential treatment advantages for a range of malignancies. A high-diversity chemical compound screen was conducted to identify agents that have the ability to alter the BRD4-dependent heterochromatization of a generic reporter in human cells. We found small molecules that mimic BRD4 inhibition without direct engagement in addition to new and known compounds that target BRD4. One such substance was a strong inhibitor of TAF1's second bromodomain. By using this inhibitor, we were able to ascertain that TAF1 functions in concert with BRD4 to regulate the growth of cancer cells, rendering TAF1 a desirable epigenetic target in MYC-driven cancers.
Animal Protocol	NA NA
References	[1]. Mapping the chemical chromatin reactivation landscape identifies BRD4-TAF1 cross-talk. Nat Chem Biol. 2016 Jul;12(7):504-10.

Solubility Data

Solubility (In Vitro)

DMSO: 23~100 mg/mL (61.7~297.3 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.43 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	2.9732 mL	14.8659 mL	29.7318 mL
	5 mM	0.5946 mL	2.9732 mL	5.9464 mL
	10 mM	0.2973 mL	1.4866 mL	2.9732 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.