NEO2734 (EP31670) is an orally bioactive, selective inhibitor of p300/CBP and BET bromodomain with IC50s <30 nM. NEO2734 is active in both SPOP mutant and wild-type prostate cancer models.

Physicochemical Properties

Molecular Formula	C22H24F3N3O3
Molecular Weight	435.439476013184
Exact Mass	435.176
CAS #	2081072-29-7
PubChem CID	126582741
Appearance	White to off-white solid powder
LogP	2.9
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	5
Heavy Atom Count	31
Complexity	733
Defined Atom Stereocenter Count	0
SMILES	FC(OCCN1C2C=C(C3C=C(C)C(N(C)C=3)=O)C=CC=2N=C1C1CCOCC1)(F)F
InChi Key	KPWWFNXRLAAREN-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H24F3N3O3/c1-14-11-17(13-27(2)21(14)29)16-3-4-18-19(12-16)28(7-10-31-22(23,24)25)20(26-18)15-5-8-30-9-6-15/h3-4,11-13,15H,5-10H2,1-2H3
Chemical Name	1,3-dimethyl-5-[2-(oxan-4-yl)-3-[2-(trifluoromethoxy)ethyl]benzimidazol-5-yl]pyridin-2-one
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	CBP/p300
ln Vitro	NEO2734 (1 μM) causes G1 cell cycle arrest and differentiation [1]. In NMC cell lines, NEO2734 (1 μM) quickly causes squamous differentiation and produces keratin or involucrin, indicators of terminal squamous differentiation [1]. In vitro and in vivo, NEO2734 exhibits activity in PCa cells harboring hotspot mutations (F133V) and non-hotspot mutations (Q165P) [2].
ln Vivo	In preclinical xenograft models, NEO2734 (5, 8, 10 mg/kg, oral) slows growth and increases survival [1].
Cell Assay	Western Blot Analysis[1] Cell Types: NUT carcinoma patient cell lines (TC-797 and PER-403). Tested Concentrations: 1 μM. Incubation Duration: 6 h. Experimental Results: Results in greater loss of MYC protein.
Animal Protocol	Animal/Disease Models: Mice (PER-403 and 14169 models)[1]. Doses: 5, 8, 10 mg/kg Route of Administration: Orally, one time/day for 28 days. Experimental Results: Two of the three mice treated with NEO2734 were alive by day 100. Provided markedly improved survival compared with EP, i-BET-762, and even iBET-762+EP by day 100 following initiation of treatment.
References	[1]. Combined Targeting of the BRD4-NUT-p300 Axis in NUT Midline Carcinoma by Dual Selective Bromodomain Inhibitor, NEO2734. Mol Cancer Ther. 2020 Jul;19(7):1406-1414. [2]. The novel BET-CBP/p300 dual inhibitor NEO2734 is active in SPOP mutant and wild-type prostate cancer. EMBO Mol Med. 2019 Nov 7;11(11):e10659.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (229.65 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.74 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 (5.74 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 (5.74 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	2.2965 mL	11.4826 mL	22.9653 mL
	5 mM	0.4593 mL	2.2965 mL	4.5931 mL
	10 mM	0.2297 mL	1.1483 mL	2.2965 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.