the most developed and best characterized medical inhibitor of Akt could be the book alkylphospholipid, perifosine. Decitabine solubility We first proved that reduction of mTOR signaling by rapamycin was related to up-regulation of Akt activation. We consequently asked whether perifosine could: prevent rapamycin induced r Akt, augment rapamycin induced cytotoxicity in vitro, and lead to increased in vivo anti-tumor activity when used with the nab based rapamycin. Our data suggests that rapamycin induced cytotoxicity was predominantly triggered as a result of autophagy in MM cells. The mix of rapamycin and perifosine led to 2 cell death inducing events: autophagy and apoptosis. Furthermore, the mixture of perifosine and nab rapamycin resulted in significant antitumor activity in an in vivo human MM cell xenograft murine model. Eventually, using the in silico predictive analysis based on a systems Extispicy biology approach we confirmed our experimental results concerning the natural effects of this drug combination. These studies consequently provide the rationale for combination clinical trials in patients with MM. MM derived cell lines Dexamethasone vulnerable MM cell line was given by Dr. Steven Rosen. The INA 6 cell line was kindly provided by Dr Martin Gramatzki. OPM1 cell line was supplied by Dr P. Leif Bergsagel. All MM cell lines were cultured in RPMI 1640 containing 100 U/mL penicillin, 2 M L glutamine, one hundred thousand fetal bovine serum, and 100 g/ mL streptomycin. After appropriate IRB approved informed consent has been previously described generation of bone-marrow stroma cells from BM specimens from MM patients obtained. Once confluent, the cells were trypsinized and passaged as needed. BMSC were incubated in 96 well culture plates for 24 hours, MM. 1S cells were then added to the wells and incubated with media alone, rapamycin, perifosine, or mixture for 48-hours at 37 C at the specified concentrations. Avagacestat solubility Rapamycin Rapamycin was obtained from Calbiochem. Perifosine Perifosine, an artificial replaced heterocyclic alkylphospholipid, was supplied by Keryx Biopharmaceuticals. nab rapamycin nab rapamycin was provided by Abraxis Bioscience LLC. Akti?? Akti?? was purchased from Calbiochem. Cell viability and proliferation assays Colorimetric assay Colorimetric assays were performed to assay drug action. Forty eight hour cultures were pulsed with 10 uL of 5 mg/mL 3 2,5 diphenyl tetrasodium bromide to each well, followed by incubation at 37 C for 4 hours, and addition of 100 uL isopropanol with 0. 04 HCl. Absorbance readings at a wavelength of 570 nm were taken on the spectrophotometer. Proliferation analysis DNA synthesis was measured by tritiated thymidine uptake as previously described.