S7= 3

S7= 3. HDAC6 (tubacin IC50 = 1.62 nM) and has a dramatically simplified synthesis (three steps, 40% overall yield). Table S1. Biochemical inhibitory activity of SAHA, WT161, and tubacin against HDAC1C9 and Fig. S2and and = 3. We previously have shown that HDAC6 inhibition by either tubacin or siRNA causes growth inhibition in MM cells (4). WT161 inhibited cell growth more potently than tubacin (Fig. 3and Fig. S2= 3. (= 4. (and Fig. S4and = 3. (= 3. Open in a separate windows Fig. S5. WT161 with BTZ induces significant cytotoxicity in patient tumor cells but not in normal PBMCs. (and indicates the assessment of WT161 vs. Tubacin (fixed concentration) in the presence of BTZ (one dose), whereas shows the combination of WT161 with BTZ at different concentrations. Moreover, these MM tumor cells are from different patients. (and and Fig. S6and Fig. S6and Fig. S7= 3. CPM, counts per minute. (= 3. (= 9 Rabbit polyclonal to SORL1 mice per group. All data represent mean SD. (= 3. (= 0.07) or WT161-treated (= 0.095) cohorts, BTZ combined with WT161 demonstrated a significant antitumor effect (= 0.0078) (Fig. 6= 0.327 and = 0.079, respectively). The on-target activity of WT161 in vivo was confirmed by assessing Ac–tubulin levels in resected tumor samples (Fig. 6and Fig. S8= 3) were injected i.v. with WT161 at 5 mg/kg. Mean plasma concentrationCtime profiles were used to calculate drug exposure [area under the curve (AUC) = 3,049 ng?L?1?h?1], t1/2 = approximately 1.4 EPZ020411 hydrochloride h, and Cmax = 18,663 ng/L. CLz, clearance; MRT, mean residence time extrapolated to infinity; t1/2z, terminal elimination half-life; Tmax, time to maximum concentration; Vz, volume of EPZ020411 hydrochloride distribution at terminal phase. ((31). Reagents and General Synthetic Procedure. Tubacin was synthesized in the J.E.B. laboratory (32). BTZ, CFZ, tubastatin A, and panobinostat were purchased from Selleck Chemicals. Antibodies used in this study were purchased directly from the vendors listed in and as previously reported by Tang et al. (33). All reactions were performed and monitored by LCMS. The intermediates and final product were fully characterized with proton and carbon-13 NMR (1H NMR and 13C NMR) spectra and high-resolution mass spectra (HRMS). Compounds were biochemically profiled against HDAC1C9 as previously reported EPZ020411 hydrochloride (14). Cell Lines. MM.1S, NCI-H929, RPMI8226, and U266 cells were obtained from American Type Culture Collection (ATCC). The KMS11 cell line was obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Lender. OPM-2 cells were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (German Collection of Microorganisms and Cell Cultures). ANBL-6 and ANBL-6-VR5 cell lines were kindly provided by Robert Orlowski, MD Anderson Cancer Center, Houston, TX. Statistical Analysis. The statistical significance of differences observed in drug-treated versus control cultures was decided using the Wilcoxon signed-ranks test. SI Materials and Methods Instrumentation. Proton and carbon-13 NMR (1H NMR and 13C NMR) spectra were recorded with a Varian inverse probe 600 INOVA spectrometer at the Harvard Medical School East Quad NMR Facility. Chemical shifts are reported in parts per million around the scale and are referenced from the residual protium in the NMR solvent (DMSO-d6: 2.50) for 1H NMR and the carbon resonances of the solvent (DMSO-d6: 40.0) for 13C NMR. Data are reported as follows: chemical shift [multiplicity (s =.