Synthesis of Inhibitors of Histone Deacet

5 4.2 P ip e r id y l SAHA A n a l o g u e ................................................................................................................1 18 4.2.1 Mono-anilide Analogue..............................................................................................118 4.2.2 Di-anilide Analogue................................................................................................... 120 4.3 M a lo n a m id e -B a s e d C o m p o u n d s .................................................................................................... 122 4.4 P ip e r id in o n e A n a l o g u e s ..................................................................................................................... 128 4.5 R e f e r e n c e s ....................................................................................................................................................133 5 BIOLOGICAL ANALYSIS............................................................................................... 134 5.1 SST4-4 A N D SST4-5.......................................................................................................134 5.1.1 Protein Analysis......................................................................................................... 135 5.1.2 Results..........................................................................................................................136 5.1.3 Cell Cycle Anaysis...................................................................................................... 141 5.2 SS581 A N D BM577.........................................................................................................147 5.3 SAHA A n a l o g u e s 150 a n d 151..................................................................................152 5.4 E x p e r i m e n t a l ............................................................................................................... 157 5.4.1 Solutions and Buffers..................................................................................................157 5.4.2 Tissue Culture..............................................................................................................157 5.4.3 Protein Analysis......................................................................................................... 157 5.4.4 Cell Cycle Analysis.....................................................................................................158 5.4.5 Histone Deacetylase Inhibition Assay......................................................................159 5 .5 R e f e r e n c e s ................................................................................................................................................... 160 6 EXPERIMENTAL..............................................................................................................161 6.1 T h e O x y g e n S e r i e s .................................................................................................................................. 162 6 .2 C y c l ic -L in k e r N o n -H y d r o x a m a t e s ............................................................................................193 6.3 A n a l o g u e s o f SAHA................................................................................................... 217 6.4 R e f e r e n c e s ...................................................................................................................................................249 6 Abbreviations and Acronyms Bn benzyl Boc terf-butoxycarbonyl BOP benzotriazol-1 -yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate Bz benzoyl CBHA m-carboxycinnamic acid bis hydroxamide DCC N,A/-dicyclohexycarbodiimide DCM dichloromethane DHAD dihydroxyacetone dimer DIPEA ethyl diisopropylamine DMAP A/,A/-dimethyl-4-aminopyridine DMEM Dulbecco's minimal essential medium DMF A/,A/-dimethylformamide DMSO dimethyl sulfoxide DNA deoxyribonucleic acid DTT dithiothreitol EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Et3N triethylamine Et20 diethyl ether EtOAc ethyl acetate EtOH ethanol FITC fluorescein isothiocyanate GF growth factor HAT histone acetyl transferase HBTU 2-(1 /-/-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HDAC histone deacetylase HMT histone methyl transferase HOBT 1-hydroxybenzotriazole HRMS high-resolution mass spectrometry HRP horse-raddish peroxidase Hz Hertz IR infra-red LHMDS lithium hexamethyldisilazide LRMS low-resoluition mass spectrometry MAPK mitogen-activated protein kinase MeOH methanol mp melting point NAD+ nicotinamide adenine dinucleotide (oxidised) NADH nicotinamide adenine dinucleotide (reduced) NBS /V-bromosuccinnimide NFF neonatal forskin fibroblast NLS nuclear-locating signal NMM A/-methylmorpholine NMR nuclear magnetic resonance NO nitric oxide PARP poly(ADP-ribose) polymerase PBS phosphate buffered solution