1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemELY 303511 hydrochlorideCat. No.: HY-15643A分式: CHClNO分量: 342.82作靶點(diǎn): TNF Receptor; Potassium Channel作通路: Apoptosis; Membrane Transporter/Ion Channel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) DM

2、SO : 45 mg/mL (131.26 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.9170 mL 14.5849 mL 29.1698 mL5 mM 0.5834 mL 2.9170 mL 5.8340 mL10 mM 0.2917 mL 1.4585 mL 2.9170 mL請(qǐng)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(qǐng)注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 LY 303511 hydroch

3、lorideLY294002 的種結(jié)構(gòu)類似物，但 LY303511 不抑制 PI3K。LY303511 可增強(qiáng)SHEP-1 神經(jīng)母細(xì)胞瘤細(xì)胞的TRAIL 敏感性。LY303511 可逆地阻斷 MIN6 胰島瘤細(xì)胞中的 K+ 電流 (IC50=64.69.1 M)。IC50 & Target TRAIL 2IC50: 64.69.1 M (K+ currents, in MIN6 insulinoma cells) 31/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 LY303511 is structurally identical t

4、o LY294002 except for a substitution of -O for -NH in the morpholine ring,and does not potently inhibit PI3K. Treatment of cells with LY303511 causes an increase in calcein spreadsimilar to levels of LY294002. The ability of LY303511 to increase gap junctional intercellular communication(GJIC) does

5、not occur concomitant with inhibition of phosphorylation of AKT as measured by immunoblotting1. LY303511 enhances TRAIL sensitivity of SHEP-1 neuroblastoma cells via H2O2-MAPK activation andup-regulation of death receptors. SHEP-1 cells are exposed to varying concentrations of LY303511 (LY30),TRAIL,

6、 and a combination of the two (1 h preincubation with LY303511 followed by TRAIL for 4 hours).SHEP-1 cells are responsive to TRAIL (10%, 15%, and 30% reduction in the surviving fraction at 25, 50,and 100 ng/mL, respectively); however, treatment with LY303511 (12.5, 25, or 50 M) has no effect on cell

7、viability. However, incubation of cells with LY303511 (25 M) for 1 hour followed by 4 hours exposure to 50ng/mL of TRAIL has a strong synergistic effect (40% reduction in viable cells with LY303511+TRAIL versus15% with TRAIL alone) 2. LY303511 is a negative control compound with respect to PI3K acti

8、vity. In MIN6insulinoma cells, Wortmannin (100 nM) has no effect on whole-cell outward K+ currents, but LY294002 andLY303511 reversibly block currents in a dose-dependent manner (IC50=9.00.7 M and 64.69.1 M,respectively). Kv2.1 and Kv1.4 are highly expressed in beta-cells, and in Kv2.1-transfected t

9、sA201 cells, 50 M LY294002 and 100 M LY303511 reversibly inhibit currents by 99% and 41%, respectively. LY303511blocks currents with an IC50 of 64.69.1 M, with a maximal inhibition of 90% at 500 M (n5 cells at eachconcentration) 3.體內(nèi)研究 Intraperitoneal administration of vehicle or LY303511 (10 mg/kg/

10、day) is performed when tumors reach avolume of 150 mm3, at which time 35 mice have developed a tumor. After 21 days, 15% of the micerequire euthanasia because of excessive tumor growth, and these data are censored due to unreliableestimates of average tumor volume. The administration of LY303511, 10

11、 mg/kg/day, is sufficient to inhibitPC-3 tumor growth in vivo 4.PROTOCOLCell Assay 2 Human neuroblastoma SHEP-1 cells are maintained in DMEM supplemented with 10% fetal bovine serumand 1% Penicillin. In a typical survival assay, SHEP-1 cells (8104 per well) plated in 24-well plates for 24 hare expos

12、ed to LY303511 (12.5, 25, and 50 M), TRAIL (25, 50, and 100 ng/mL), and a combination of thetwo (1 h preincubation with LY303511 followed by TRAIL for 4 h). Cytotoxicity is determined by the crystalviolet assay. After drug exposure, cells are washed with PBS and incubated for 20 min with crystal vio

13、letsolution (200 L). The excess crystal violet solution is washed away with distilled water, and the remainingcrystals are dissolved with 20% acetic acid. Viability is determined by absorbance at 595 nm wavelengthusing an automated ELISA reader. Cell viability experiments are performed similarly wit

14、h 2,000 units/mL ofcatalase, 4 M JNK inhibitor SP600125, 10 M p38 inhibitor SB202190, 20 M MAPK/ERK kinase (MEK)inhibitor PD98059, 50 M of caspase-8 inhibitor Z-IETD-FMK or pan-caspase inhibitor Z-VAD-FMK, or deathreceptor blocking antibodies (4 g/mL anti-DR4 or 1 g/mL anti-DR5), or in cells transfe

15、cted with smallinterfering RNA (siRNA) for silencing JNK and ERK expression, respectively. Cells are preincubated for 1 hwith LY303511 and the respective inhibitor or catalase before the addition of TRAIL. Similar sensitizing effectof LY303511 on TRAIL-induced apoptosis is carried out with SY5Y neur

16、oblastoma, T98G glioblastoma,Jurkat leukemia, CEM myelogenous leukemia, HeLa ovarian carcinoma, and HT29 colorectal carcinoma celllines 2.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEMCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 4Admin

17、istration 4 Human prostate adenocarcinoma (PC-3) cells (ATCC CRL-1435) are cultured in vitro before harvesting andimplantation of 1106 cells in 20% Matrigel per athymic NCR nude mouse by subcutaneous injection at theflank. Inoculated mice are subdivided into four groups of 10. Administration of vehi

18、cle or LY303511, 10mg/kg/day, is begun (day 1) when tumors reach 150 mm3 (n=35), and tumor volumes are measured for 30days at the indicated time points.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Bodenstine TM, et al. Homotypic gap junc

19、tional communication associated with metastasis suppression increases with PKA activity andis unaffected by PI3K inhibition. Cancer Res. 2010 Dec 1;70(23):10002-11.2. Shenoy K, et al. LY303511 enhances TRAIL sensitivity of SHEP-1 neuroblastoma cells via hydrogen peroxide-mediated mitogen-activated protein kinase activation and up-regulation of death receptors. Cancer Res. 2009 Mar 1;69(5):1941-50.3. El-Kholy W, Macdonald PE