ZM 336372

c-Raf (IC50 = 0.07 μM)

ZM 336372 shows 10-fold selectivity over B-Raf. ZM 336372 selectively inhibits 17 other protein kinases, including PKA, PKC, AMPK, p42 MAPK, MKK1, SAPK1/JNK, and CDK1, even at concentrations as high as 50 μM.It weakly inhibits SAPK2a/p38α and SAPK2b/p38β with an IC50 of 2 μM. ZM 336372 does not stop the activation of MKKl or p42 MAPK/ERK2 that is caused by constitutive, growth factor, or phorbol ester induction. Additionally, ZM 336372 does not change the phenotype of cell lines with Ras- or Raf-transformed. ZM 336372 treatment causes >100 activation of c-Raf and the B-Raf isoform but no activation of MKKI, p42 MAPK/ERKP, or any increase in the GTP-loading of Ras, indicating the existence of a feedback control loop by which Raf isoforms suppress their own activation, such that inhibition is always counterbalanced by reactivation. Inhibiting the MAPK cascade, protein kinase C, or phosphatidylinositide 3-kinase does not stop ZM 336372-induced activation of c-Raf. [1] After exposure to hydrogen peroxide, eNOS upregulation is eliminated by ZM 336372 (1 M).[2] In carcinoid tumor cells, treatment with ZM 336372 causes Raf-1, mitogen-activated protein kinase 1/2, and extracellular signal-regulated kinase 1/2 to become increasingly phosphorylated. It also significantly lowers levels of bioactive hormones and the transcription factor human achaete-scute homologue-1. Additionally, ZM 336372 treatment results in the induction of the cell cycle inhibitors p21 and p18 as well as a notable suppression of cellular proliferation. [3] Pheochromocytoma cell proliferation is inhibited by ZM 336372, and NE vasoactive peptide production is suppressed.[4] Treatment with ZM 336372 in HepG2 results in the up-regulation of cell cycle inhibitors, a dose-dependent suppression of proliferation, and a suppression of hormone secretion.[5] By blocking glycogen synthase kinase-3β and phosphorylating GSK-3β at Ser 9, ZM 336372 also causes apoptosis in pancreatic adenocarcinoma cell lines. [6]

N/A

Sl9 cell lysates are used to directly measure the activity of c-Raf kinase. In the absence of ZM 336372, cotransfection of baculovirus vectors containing DNA encoding v-Ras and Lck activates human c-Raf in Sf9 cells. Then, with increasing concentrations of ZM 336372, the cell lysates are tested for c-Raf activity.

ZM 336372 is applied to cells at different concentrations for 48 and 72 hours. Cells are trypsinized after the medium has been removed from the incubation. Prior to flow cytometry, cells are incubated on ice for 5 minutes and 2.5 g/mL propidium iodide is added. CellQuest acquisition and analysis software is used to collect data using a FACSCalibur benchtop flow cytometer. Cell Titer Glo Assay is used to measure cytotoxicity. Using the MTT assay, cell proliferation is measured.

[1]. Chem Biol . 1999 Aug;6(8):559-68.

[2]. Int J Cancer . 2003 Apr 10;104(3):274-82.

[3]. Mol Cancer Ther . 2005 Jun;4(6):910-7.

[4]. J Surg Res . 2006 Jun 1;133(1):42-5.

[5]. J Gastrointest Surg . 2008 May;12(5):852-7.

[6]. J Surg Res . 2010 Jun 1;161(1):28-32.

C23H23N3O3

389.45

389.17

C, 70.93; H, 5.95; N, 10.79; O, 12.32

208260-29-1

Solid powder

CC1=C(C=C(C=C1)NC(=O)C2=CC(=CC=C2)N(C)C)NC(=O)C3=CC=C(C=C3)O

PYEFPDQFAZNXLI-UHFFFAOYSA-N

InChI=1S/C23H23N3O3/c1-15-7-10-18(24-23(29)17-5-4-6-19(13-17)26(2)3)14-21(15)25-22(28)16-8-11-20(27)12-9-16/h4-14,27H,1-3H3,(H,24,29)(H,25,28)

3-(Dimethylamino)- N -[3-[(4-hydroxybenzoyl)-amino]-4-methylphenyl]benzamide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)