ABSTRACT
Guided by structure-based design, we synthesized two novel series of potent inhibitors of BACE1 and generated extensive SAR around both the prime and non-prime side binding pockets. The key feature of both series is a cyclic amine motif specifically crafted to achieve interactions with both the flap and with the S2' pocket.
Subject(s)
Amyloid Precursor Protein Secretases/antagonists & inhibitors , Aspartic Acid Endopeptidases/antagonists & inhibitors , Drug Design , Imidazolidines/chemical synthesis , Imidazolidines/pharmacology , Piperazines/chemical synthesis , Piperazines/pharmacology , Animals , Crystallography, X-Ray , Disease Models, Animal , Humans , Imidazolidines/chemistry , Mice , Mice, Transgenic , Models, Molecular , Molecular Conformation , Molecular Structure , Piperazines/chemistry , Structure-Activity RelationshipABSTRACT
The examination of a key series of chromophore analogues of sandramycin (1) is detailed employing surface plasmon resonance to establish binding constants within a single high affinity bis-intercalation binding site 5'-d(GCATGC)2, and to establish the preference for sandramycin binding to 5'-d(GCXXGC)2 where XX=AT, TA, GC, and CG. From the latter studies, sandramycin was found to exhibit a preference that follows the order: 5'-d(GCATGC)2 > 5'-d(GCGCGC)2, delta deltaG(o)= 0.4 kcal/mol > 5'-d(GCTAGC)2, delta deltaG(o) = 0.9 kcal/mol> or =5'-d(GCCGGC)2, delta delta G(o) = 1.0 kcal/mol although it binds with high affinity to all four deoxyoligonucleotides. The two highest affinity sequences constitute repeating 5'-PuPy motifs with each intercalation event occurring at a 5'-PyPu step. The most effective sequence constitutes the least stable duplex, contains the sterically most accessible minor groove central to the bis-intercalation site, and the ability to accept two gly-NH/T C2 carbonyl H-bonds identified in prior NMR studies. Similarly, the contribution of the individual structural features of the chromophore were assessed with the high affinity duplex sequence 5'-d(GCATGC)2. In addition to the modest affinity differences, one of the most distinguishing features of the high affinity versus lower affinity bis-intercalation or mono-intercalation directly observable by surface plasmon resonance was the temporal stability of the complexes characterized by the exceptionally slow off-rates.
Subject(s)
DNA-Binding Proteins/chemistry , DNA/chemistry , Kinetics , Models, Chemical , Peptides, Cyclic/chemistry , Spectrometry, Fluorescence , Surface Plasmon Resonance , Time FactorsABSTRACT
The preparation and examination of 2-22 constituting a systematic study of the chromophore of sandramycin (1) are detailed. Fluorescence quenching studies were used to establish binding constants for 1-24 within calf thymus DNA, within a single high affinity bis-intercalation binding site 5'-d(GCATGC)2, and to establish the preference for sandramycin binding to 5'-d(GCXXGC)2 where XX = AT, TA, GC, and CG. From the latter studies, sandramycin was found to exhibit a preference that follows the order: 5'-d(GCATGC)2 > 5'-d(GCGCGC)2, deltadeltaGo = 0.3kcal/mol > 5'-d(GCTAGC)2, 5'-d(GCCGGC)2, deltadeltaGo = 0.6 kcal/mol although it binds with high affinity to all four deoxyoligonucleotides. The two highest affinity sequences constitute repeating 5'-PuPy motifs with each intercalation event occurring at a 5'-PyPu step. The most effective sequence constitutes the less stable duplex, contains the sterically most accessible minor groove central to the bis-intercalation site, and the ability to accept two gly-NH/TC2 carbonyl H-bonds identified in prior NMR studies. Similarly, the contribution of the individual structural features of the chromophore were assessed with the high affinity duplex sequence 5'-d(GCATGC)2. To a first approximation, the cytotoxic properties were found to parallel trends established in the DNA binding affinities. The exception to this generalization was 4 which lacks the sandramycin chromophore phenol. Although typically 4-10x less potent than sandramycin against leukemia cell lines, it proved to be 1-10,000x more potent against melanomas, carcinomas, and adenocarcinomas exhibiting IC50 values of 1 pM-10 nM placing it among the most potent agents identified to date. Additionally, the first disclosure of the HIV-1 reverse transcriptase inhibitory activity of sandramycin (1) as well as that of its key analogs are described and define the chromophore structural features required for their exceptional potency. Two analogs, 18 and 3, roughly maintain the HIV-1 reverse transcriptase inhibitory potency of 1 but exhibit substantially diminished cytotoxic activity (10(2)-10(3)x).
Subject(s)
Antibiotics, Antineoplastic/pharmacology , DNA/drug effects , Intercalating Agents/pharmacology , Animals , Antibiotics, Antineoplastic/chemical synthesis , Binding Sites , Cattle , Cell Death , DNA/metabolism , Female , HIV Reverse Transcriptase/antagonists & inhibitors , Humans , Hydroxyquinolines/pharmacology , Intercalating Agents/chemical synthesis , Mice , Oligonucleotides/metabolism , Peptides, Cyclic/chemical synthesis , Peptides, Cyclic/metabolism , Peptides, Cyclic/pharmacology , Phenol/chemistry , Spectrometry, Fluorescence , Structure-Activity Relationship , Tumor Cells, Cultured/drug effectsABSTRACT
The synthesis of hosts with improved binding affinities for nitroaromatic guests is described. Association constants for several host-guest complexes were measured in chloroform solution and ranged over three orders of magnitude. Two hosts were covalently linked to silica gel to produce chemically bonded stationary phases for HPLC. The use of these phases for HPLC analysis of nitro-substituted polycyclic aromatic hydrocarbons is discussed.