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Both mackinazolinone (5) and isaindigotone (6) only demonstrated cytotoxicity at > 100 μM concentrations in the selected cancer cell lines in the MTS assay described above. For primary screening, our activity cutoff was ≤50 μM.
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For the design of libraries possessing maximal information, content, and diversity, see: Patterson, J. E.; Zhang, Y.; Smellie, A.; Li, D.; Hartsough, D. S.; Yu, L.; Baldino, C. M. Abstracts of Papers, 229th ACS National Meeting, San Diego, CA, March 13-17, 2005, CINF-75.
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We employed a Biotage Smith Synthesizer integrated into the ArQule AMAP high-throughput parallel chemistry platform.
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The purification of crude products were performed on the Preparative HPLC. The preparative HPLCs were conducted using a ProntoSIL 120-10-C18 column (50 × 20 mm). The flow rate was at 40 mL/min utilizing an acetonitrile/water mobile phase and 0.1% trifluoroacetic acid as a modifier with mass-triggered fraction collection. The crude and purified products were characterized by LC/MS with an integrated LC/MS system of a Shimadzu LC instrument and Micromass DMZ MS instrument. The HPLC was conducted using a Zorbax SB-C8 column (4.6 × 30 mm 3.5μm) with a flow rate at 3.0 mL/min, an acetonitrile/water mobile phase, and 0.05% trifluoroacetic acid as a modifier. For detailed information, see: (a) Kyranos, J. N.; Cai, H.; Zhang, B.; Goetzinger, W. K. Curr. Opin. Drug Discovery Dev. 2001, 4, 719.
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The three cancer cell lines represent diverse tissue sources. Two sets of assays (n = 2) were run with 20 data points using Taxol as a positive control.
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