CGK-733 905973-89-9_ATM(ATR)_DNA Damage_Signaling Pathways_Products

CGK 733 is a novel, potent and selective inhibitor of ATM (ataxia-telangiectasia mutated) and ATR (ATM and Rad3-related) with the potential to treat Hepatocellular carcinoma (HCC). Its IC50 is around 200 nM, and it inhibits ATM/ATR. The cytotoxicity of taxol induced in HBV-positive HepG2.2.15 cells was markedly increased by CGK733. CGK733 may offer a novel treatment approach for patients with HBV-infected HCC and may be able to reverse the taxol resistance in HBV-positive HCC cells.

Physicochemical Properties

Molecular Formula

C23H18CL3FN4O3S

Molecular Weight

555.84

Exact Mass

554.014

Elemental Analysis

C, 49.70; H, 3.26; Cl, 19.13; F, 3.42; N, 10.08; O, 8.64; S, 5.77

CAS #

905973-89-9

Related CAS #

905973-89-9

PubChem CID

6605258

Appearance

White to off-white solid powder

Density

1.5±0.1 g/cm3

Index of Refraction

1.674

LogP

6.55

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

725

Defined Atom Stereocenter Count

SMILES

O=C(C(C1C=CC=CC=1)C1C=CC=CC=1)NC(C(Cl)(Cl)Cl)NC(NC1C=C([N+](=O)[O-])C(F)=CC=1)=S

InChi Key

HLCDNLNLQNYZTK-UHFFFAOYSA-N

InChi Code

InChI=1S/C23H18Cl3FN4O3S/c24-23(25,26)21(30-22(35)28-16-11-12-17(27)18(13-16)31(33)34)29-20(32)19(14-7-3-1-4-8-14)15-9-5-2-6-10-15/h1-13,19,21H,(H,29,32)(H2,28,30,35)

Chemical Name

2,2-diphenyl-N-[2,2,2-trichloro-1-[(4-fluoro-3-nitrophenyl)carbamothioylamino]ethyl]acetamide

Synonyms

CGK733; CGK-733; CGK 733

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

ATM; ATR

ln Vitro

In vitro activity: CGK733 (4.2 ng/μL-12.5 ng/μL) amplifies the cytotoxicity caused by taxol in HBV-positive HCC cells. CGK733 (4.2 ng/μL) stimulates the exit of mitosis and speeds up the production of multinucleated cells in taxol-treated HBV-positive HCC cells[1]. In breast cancer cell lines T47D and MCF-7, CGK733 (10 μM) causes cyclin D1 to be lost via the ubiquitin-dependent proteasomal degradation pathway. Exposure to CGK733 (0.6–40 μM) inhibits the proliferation of LnCap prostate cancer cells, HCT116 colon cancer cells, MCF-7 and T47D estrogen receptor positive breast cancer cells, and MDA-MB436 ER negative breast cancer cells. Additionally, CGK733 prevents the growth of embryonic fibroblast cells from non-transformed mice, BALB/c 3T3. Moreover, pan-caspase inhibition is unable to counteract the proliferation-inhibiting effect of CGK733 (10 μM) on MCF-7[2]. In HEK-293 cells, CGK733 (10 μM) causes a 1.6-fold increase in ATM reporter activity[3].

ln Vivo

CGK733 (25 mg/kg, i.p.) enhances the ATM reporter activity (which reports the deactivation of ATM kinase activity) with changes of 2.4, 3.1, and 1.3 fold at 1, 4, and 8 hours, respectively, in comparison to control mice[3].

Cell Assay

For the assay to be completed with exponential growth, cells are seeded in 96-well plates at a predefined optimal cell density. The growth medium is swapped out for experimental medium containing the right amounts of the drugs or a 0.1% (v/v) vehicle control after a 24-hour preincubation. The assay for measuring sulforhodamine B colorimetric cell proliferation is used after a 48-hour incubation period. The resultant expression is the mean ± SE for six replicates expressed as a percentage of vehicle control, which is represented as 100%. At least three separate experiments are conducted. The Student's t test, with two tails, is used for statistical analyses. Statistics are defined as significant when P < 0.05[2].

Animal Protocol

A minimum of one week is spent acclimating athymic CD-1 female mice, aged four to six weeks, prior to use. Two times as many D54-ATMR cells are subcutaneously injected into each flank of the mice. It is permissible for tumors to reach sizes of 100–150 mm³. Intraperitoneal injections of vehicle control (DMSO), CGK-733, KU-55933 (25 mg/kg), or 5 Gy of radiation are administered to each flank of the mice. After injecting 400 μg/100 μL of D-luciferin at baseline (-3h) and 1, 4, and 8 hours after drug administration, bioluminescence is recorded on the Xenogen IVIS Spectrum system[3].

References

[1]. CGK733 enhances multinucleated cell formation and cytotoxicity induced by taxol in Chk1-deficient HBV-positive hepatocellular carcinoma cells. Biochem Biophys Res Commun. 2012 May 25;422(1):103-8.

[2]. The ATM and ATR inhibitors CGK733 and caffeine suppress cyclin D1 levels and inhibit cell proliferation. Radiat Oncol. 2009 Nov 10;4:51.

[3]. Molecular imaging of the ATM kinase activity. Int J Radiat Oncol Biol Phys. 2013 Aug 1;86(5):969-77.

Additional Infomation

2,2-diphenyl-N-[2,2,2-trichloro-1-[[(4-fluoro-3-nitroanilino)-sulfanylidenemethyl]amino]ethyl]acetamide is a diarylmethane.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~179.9 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.50 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.

Solubility in Formulation 2: ≥ 2.08 mg/mL (3.74 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 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.

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7991 mL	8.9954 mL	17.9908 mL
	5 mM	0.3598 mL	1.7991 mL	3.5982 mL
	10 mM	0.1799 mL	0.8995 mL	1.7991 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.

PeptideDB

CGK-733 905973-89-9

CAS No.: 905973-89-9

Physicochemical Properties

Biological Activity

Solubility Data