1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemELY3009120Cat. No.: HY-12558CAS No.: 1454682-72-4Synonyms: DP-4978分式: CHFNO分量: 424.51作靶點(diǎn): Raf; Autophagy作通路: MAPK/ERK Pathway; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 38 mg/

2、mL (89.51 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.3557 mL 11.7783 mL 23.5566 mL5 mM 0.4711 mL 2.3557 mL 4.7113 mL10 mM 0.2356 mL 1.1778 mL 2.3557 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 LY3009120是泛 RAF 抑制劑，其抑制BRAFV6

3、00E，BRAFWT 和CRAFWT 的 IC50 分別為5.8，9.1和15 nM。IC50 & Target BRafV600E Braf CRAF5.8 nM (IC50) 9.1 nM (IC50) 15 nM (IC50)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 In the whole-cell based KiNativ assay, LY3009120 shows affinity to each RAF isoform with the IC50 of 44,31-47 and 42 nM for ARAF,

4、BRAF and CRAF respectively. LY3009120 exhibits anti-proliferative effects oncell lines harboring BRAFV600E, KRASG13 and KRASG12 mutations. LY3009120 (1 M) inhibits thephosphorylation of both MEK1/2 and ERK1/2 in cell lines with high basal levels of pMEK1/2 and pERK1/2(RKO and HCT 116) 1. LY3009120 s

5、hows inhibitory effect on tumor cells such as BxPC-3, NCI-H2405 andOV-90 cell lines. LY3009120 (0.01 M) demonstrates potent and dose-dependent inhibition of phospho-MEKand ERK in all three cell lines. LY3009120 demonstrates a concentration-dependent cell growth inhibitionwith IC50 values of 0.04, 0.

6、087, and 0.007 M against H2405, BxPC-3, and OV-90 cells, respectively 2.LY3009120 inhibits BRAFWT, CRAFWT, BRAFV600E, and BRAFV600E+G468A with the IC50 values of 9.1,15, 5.8, and 17 nM, respectively. LY3009120 induces BRAF-CRAF dimerization but inhibits thephosphorylation of downstream MEK and ERK.

7、LY3009120 also inhibits various forms of RAF dimersincluding BRAF or CRAF homodimers 3. LY3009120 gives only very minor activation at very low doses,with near complete inhibition of phospho-ERK at concentrations above 100 nM 4.體內(nèi)研究 LY3009120 (20 mg/kg bid) displays significant activity in in vivo BR

8、AFmut and KRASmut CRC xenograftmodels. In Colo 205 xenografts (BRAFmut), LY3009120 results in statistically significant tumor regression,while treatment of HCT 116 xenografts (KRASmut) results in statistically significant inhibition of tumor growth.LY3009120 treatment reduces pMEK1/2 in all HT-29 xe

9、nografts and reduces pERK1/2 in the majority of HT-29 xenografts 1. LY3009120 (15 or 30 mg/kg) achieves almost complete tumor growth regression, andinhibits downstream phospho-MEK and ERK by approximately 70% and 60%, respectively, in the H2405model 2.PROTOCOLKinase Assay 3 In the BRAF WT enzymatic

10、assay, the reaction mixture contains 1.2 nM BRAF, 30 nM MEK1, 1000 M ATP,3.5 units (per 100 L) of PK, 5 units (per 100 L) of LDH, 1 mM PEP, and 280 M of NADH. In the CRAFassay, the reaction mixture contains 0.6 nM CRAF, 26 nM MEK1, 2000 M ATP, and the same amount ofPK, LDH, PEP and NADH. In the BRAF

11、V600E assay, the reaction mixture contains 1.6 M BRAFV600E, 26nM MEK1, 200 M ATP and the same amount of PK, LDH, PEP and NADH as above. In theBRAFV600E+G468A assay, the reaction mixture contains 3.5 nM BRAF, 30 nM MEK1, 200 M ATP and thesame amount of PK, LDH, PEP and NADH as above. All assays are s

12、tarted by mixing the above mixture withtest compound and monitored at A340 continuously for approximately 5 hr. Reaction data at the 3 to 4 hourtime frame are collected to calculate IC50.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 4 Brief

13、ly, cells are grown in McCoys 5A supplemented with 10% characterized fetal bovine serum and 1%penicillin/streptomycin/l-glutamine at 37C, 5% CO2, and 95% humidity. Cells are allowed to expand until 75-90% confluency at which point they are subcultured or harvested for assay use. A serial dilution of

14、 testcompound is dispensed into a 384-well black clear bottom plate in triplicate. Six-hundred-twenty-five cells areadded per well in 50 L of complete growth medium in the 384-well plate. Plates are incubated for 67 h at37C, 5% CO2, and 95% humidity. At the end of the incubation period, 10 L of a 44

15、0 M solution ofresazurin in PBS is added to each well of the plate and plates are incubated for an additional 5 h at 37C, 5%CO2, and 95% humidity. Plates are read on a Synergy2 reader using an excitation of 540 nm and an2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEemission of 600 nm. Data are an

16、alyzed using Prism softwareto calculate IC50 values.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Briefly, 5106 to 10106 tumor cells in a 1:1 Matrigel mix (0.2 mL total volume) are injected subcutaneouslyAdministration 2 into the right hind fla

17、nk of female NIH nude rats. After tumors reach a desired size of approximately 300mm3, animals are randomized into groups of 8 for efficacy studies. Drugs (LY3009120 or vemurafenib) areadministered orally (gavage) in 0.6-mL volume of vehicle with the dose schedules. Tumor growth and bodyweight are m

18、onitored over time to evaluate efficacy and signs of toxicity.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Oncogene. 2018 Oct;37(43):5719-5734. Clin Sci (Lond). 2019 Apr 16;133(8):919-932. Cancer Sci. 2018 Jan;109(1):121-131. Methods Mol

19、Biol. 2018;1711:351-398.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Vakana E, et al. LY3009120, a panRAF inhibitor, has significant anti-tumor activity in BRAF and KRAS mutant preclinical models ofcolorectal cancer. Oncotarget. 2017 Feb 7;8(6):9251-92662. Chen SH, et al. Oncogenic BRAF Deletions That Function as Homodimers and Are Sensitive to Inhibition by RAF Dimer InhibitorLY3009120. Cancer Discov. 2016 Mar;6(3):