RAF709

製品コードS8690 バッチS869001

印刷

化学情報

Synonyms

N/A

Storage
(From the date of receipt)

3 years -20°C powder
1 years -80°C in solvent

化学式

C₂₈H₂₉F₃N₄O₄

分子量

542.55

CAS No.

1628838-42-5

Solubility (25°C)*

体外

DMSO

100 mg/mL (184.31 mM)

Ethanol

100 mg/mL (184.31 mM)

Water

Insoluble

体内（毎回新しく調製した物を用意してください）

Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O この製剤はselleckのラボで検証済みです。上記の溶解方法がご要望を満たさない場合、selleckの営業担当までお問い合わせ頂ければ、個別の試験を行います。	5.000mg/ml (9.22mM)	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil この製剤はselleckのラボで検証済みです。上記の溶解方法がご要望を満たさない場合、selleckの営業担当までお問い合わせ頂ければ、個別の試験を行います。	0.800mg/ml (1.47mM)	Taking the 1 mL working solution as an example, add 50 μL of 16 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明	RAF709 is a potent inhibitor of B/C RAF kinase with almost equivalent IC50 values of 0.4 nM for B-RAF and C-RAF, showing a high level of selectivity, demonstrating greater than 99% on-target binding to BRAF, BRAFV600E, and CRAF at 1 μM and very few off-targets with DDR1 (>99%), DDR2 (86%), FRK (92%), and PDGFRb (96%), the only kinases with binding >80% at 1 μM.
in vitro	RAF709 appears to have very slow dissociation kinetics (T_1/2 > 6.5 h) using the rapid dilution method to measure its dissociation rate constant. In cellular assays, the dose−response of pMEK and pERK are measured in Calu-6 cells with EC50 of 0.02 and 0.1 μM with minimal paradoxical activation and inhibition of proliferation with EC50 of 0.95 μM. This compound stabilizes BRAF−CRAF dimers with an EC50 of 0.8 μM. Of the 456 kinases tested, this chemical shows a high level of selectivity, demonstrating greater than 99% on-target binding to BRAF, BRAFV600E, and CRAF at 1 μM and very few off-targets with DDR1 (>99%), DDR2 (86%), FRK (92%), and PDGFRb (96%), the only kinases with binding >80% at 1 μM^[1]. It shows equal activity against both RAF monomers and dimers. In in vitro biochemical assays, this compound exhibits potent inhibitory activity targeting BRAF, BRAFV600E, and CRAF with IC50 values ranging between 0.3 to 1.5 nmol/L. This chemical treatment leads to a dose-dependent induction of B/CRAF heterodimerization in HCT116, but inhibits MEK and ERK phosphorylation, in line with the ability of this compound to effectively inhibit the RAF dimers. It selectively inhibits oncogenic signaling and proliferation in tumor cells with BRAF, NRAS, or KRAS mutations with minimal paradoxical activation^[2].
in vivo	RAF709 is well tolerated and efficacious in KRAS mutant xenograft models. It is reasonably stable in plasma after a 3 h incubation at 37℃ across species [plasma stability (%remaining): rat 85%, mouse 82%, dog 95%, human 101%], and plasma protein binding is measured to be 98% across species. In pharmacokinetic experiments, this compound has moderate clearance in mouse (35 mL/min/kg) and dog (14 mL/min/kg) and high clearance in rat (50 mL/min/kg). Cmax in mouse (1 μM), dog (0.5 μM), and rat (0.5 μM) reach pharmacologically active concentrations, and acceptable oral availability is observed in mouse (68%), rat (24%), and dog (48%). In the Calu-6 xenograft nude mouse model, treatment with this chemical results in dose-dependent antitumor activity with 10 mg/kg being subefficacious (%T/C = 92%), 30 mg/kg resulted in measurable antitumor activity (% T/C = 46%), and 200 mg/kg resulted in mean tumor regression of 92%, while the same high dose is not efficacious in the PC3, KRAS WT mode^[1].

プロトコル(参考用のみ)

キナーゼアッセイ	CRAF kinase assay
キナーゼアッセイ	The CRAF kinase assay was carried out using 10 nM kinase-dead MEK1 protein substrate (carrying a K97R mutation), 3 μM ATP, and 10 pM CRAF Y340E/Y341E. The reaction buffer contained 50 mM Tris pH 7.5, 10 mM MgCl2, 0.05% BSA, 50 mM NaCl, 0.01% Tween-20, and 1 mM DTT. The reactions were carried out at room temperature in a volume of 10 μL in white 384-shallow-well plates for 40 min and stopped by adding 5 μL/well quench solution (50 mM Tris pH 7.5, 50 mM EDTA). Terminated reactions received 5 μL/well detection reagents consisting of 50 mM Tris pH 7.5, 0.01% Tween-20, 1:1000 diluted antiphospho MEK1/2 S217/S221 antibody, 0.01 mg/mL each of AlphaScreen Protein A-coated acceptor beads, and streptavidin-coated donor bead. Plates were read in an EnVision plate reader after overnight incubation at room temperature. In compound inhibition studies, this compound was tested over a concentration range of 25 μM to 1.74 × 10⁻⁶ μM in 16-point, 3-fold format. DMSO was at a final concentration of 0.5%. This chemical was preincubated with CRAF for 30 min before adding substrates to start the reaction. Inhibition data were fit to a four-parameter logistic equation to calculate the IC50 of the compounds.
細胞アッセイ	細胞株	HCT116 cells
	濃度	0.1, 1, 10 μM
	反応時間	1 hour
	実験の流れ	BRAF/CRAF dimerization is assessed by immunoprecipitating BRAF or CRAF, followed by Western blot analysis of BRAF and CRAF. Levels of pMEK and pERK in whole-cell lysates (WCL) are determined by Western blot analysis. GAPDH level is included as a loading control.
動物実験	動物モデル	Calu-6 model (tumor bearing mice)
	投薬量	10, 30, or 200 mg/kg
	投与方法	oral administration

参考

https://pubmed.ncbi.nlm.nih.gov/28557458/
https://pubmed.ncbi.nlm.nih.gov/29343524/

Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

Tumour-selective activity of RAS-GTP inhibition in pancreatic cancer [ Nature, 2024, 629(8013):927-936]	PubMed: 38588697
Identification of a RAS-activating TMEM87A-RASGRF1 fusion in an exceptional responder to sunitinib with non-small cell lung cancer. [ Clin Cancer Res, 2020, 20 pii: clincanres]	PubMed: 32312893
KRASQ61H preferentially signals through MAPK in a RAF dimer-dependent manner in non-small cell lung cancer [ Cancer Res, 2020, canres.0448.2020]	PubMed: 32605999
Depolarization-Dependent C-Raf Signaling Promotes Hyperexcitability and Reduces Opioid Sensitivity of Isolated Nociceptors after Spinal Cord Injury [ J Neurosci, 2020, 40(34):6522-6535]	PubMed: 32690613

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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