CCT020312 (CCT-020312) is a novel and potent EIF2AK3 activator that sensitizes cancer cells with defective taxane-induced EIF2A phosphorylation to paclitaxel treatment.

Physicochemical Properties

Molecular Formula	C31H30BR2N4O2
Molecular Weight	650.403505802155
Exact Mass	648.073
CAS #	324759-76-4
PubChem CID	3108148
Appearance	Light yellow to yellow solid powder
LogP	5.6
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	8
Heavy Atom Count	39
Complexity	950
Defined Atom Stereocenter Count	0
SMILES	BrC1C=CC(=CC=1)C1CC(C2C(NC3C=CC(=CC=3C=2C2C=CC=CC=2)Br)=O)=NN1C(CCN(CC)CC)=O
InChi Key	OAXDKXMGESZLKV-UHFFFAOYSA-N
InChi Code	InChI=1S/C31H30Br2N4O2/c1-3-36(4-2)17-16-28(38)37-27(20-10-12-22(32)13-11-20)19-26(35-37)30-29(21-8-6-5-7-9-21)24-18-23(33)14-15-25(24)34-31(30)39/h5-15,18,27H,3-4,16-17,19H2,1-2H3,(H,34,39)
Chemical Name	6-bromo-3-[3-(4-bromophenyl)-2-[3-(diethylamino)propanoyl]-3,4-dihydropyrazol-5-yl]-4-phenyl-1H-quinolin-2-one
Synonyms	CCT 020312 CCT-020312 CCT020312
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	EIF2AK3[1]. PERK[2].
ln Vitro	Upon subjecting HT29 cells to CCT020312 for a whole day, a concentration-dependent decrease in P-S608-pRB signal was observed, exhibiting a linear response ranging from 1.8 to 6.1 μM [1]. Even while CCT020312 treatment only lasted for two hours and the substance was later removed, it successfully suppressed cell proliferation (measured at 96 hours), suggesting that CCT020312 is capable of eliciting long-lasting rather than temporary cellular inhibition [1]. After subjecting HT29 cells to a 24-hour treatment of 10 μM CCT020312, the amount of G1/S cyclins D1, D2, E, and A as well as the CDK catalytic subunit CDK2 were reduced. Conversely, the levels of the CDK inhibitor p27KIP1 were elevated in these cells [1].
ln Vivo	Upon administering the PERK activator CCT020312 (1–5 mg/kg; intraperitoneally; once daily for three days) to 15-week-old wild-type mice, brain homogenates demonstrated elevated levels of phosphorylated PERK and NRF2 [2]. In the Morris water maze, P301S transgenic mice given CCT020312 (2 mg/kg; i.p.; once daily for 6 weeks) fared far better [2].
Animal Protocol	Animal/Disease Models: 9weeks old P301S tau transgenic mice[2] Doses: 2 mg/kg Route of Administration: intraperitoneal (ip)injection; one time/day for 6 weeks Experimental Results: P301S transgenic mice treated with CCT020312 performed Dramatically better in Morris water maze.
References	[1]. Mechanism-based screen for G1/S checkpoint activators identifies a selective activator of EIF2AK3/PERK signalling. PLoS One. 2012;7(1):e28568. [2]. PERK activation mitigates tau pathology in vitro and in vivo. EMBO Mol Med. 2017 Mar;9(3):371-384.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~153.75 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.20 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 (3.20 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 (3.20 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.5375 mL	7.6876 mL	15.3752 mL
	5 mM	0.3075 mL	1.5375 mL	3.0750 mL
	10 mM	0.1538 mL	0.7688 mL	1.5375 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.