ABSTRACT
Upon binding, ligands can chaperone their protein targets by preventing them from misfolding and aggregating. Thus, an organic molecule that works as folding chaperone for a protein might be its specific ligand, and, similarly, the chaperone potential could represent an alternative readout in a molecular screening campaign toward the identification of new hits. Here we show that small molecules selected for acting as pharmacological chaperones on a misfolded mutant of the Frizzled4 (Fz4) receptor bind and modulate wild-type Fz4, representing what are to our knowledge the first organic ligands of this until-now-undruggable GPCR. The novelty and the advantages of the screening platform, the allosteric binding site addressed by these new ligands and the mechanism they use to modulate Fz4 suggest new avenues for development of inhibitors of the Wnt-ß-catenin pathway and for drug discovery.
Subject(s)
Frizzled Receptors/chemistry , Molecular Chaperones/chemistry , Allosteric Site , Amino Acid Motifs , Base Sequence , Cell Line, Tumor , Chemistry, Pharmaceutical/methods , Dose-Response Relationship, Drug , Drug Design , Glycerol/chemistry , HEK293 Cells , HeLa Cells , Humans , Ligands , Microscopy, Fluorescence , Molecular Sequence Data , Mutagenesis , Protein Binding , Protein Folding , Receptors, G-Protein-Coupled/chemistryABSTRACT
The first direct activator of BAX, a pro-apoptotic member of the BCL-2 family, has been recently identified. Herein, a structure-based lead optimization turned out into a small series of analogues, where 8 is the most potent compound published so far. 8 was used as pharmacological tool to ascertain, for the first time, the anticancer potential of BAX direct activators and the obtained results would suggest that BAX direct activators are potential future anticancer drugs rather than venoms.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Drug Screening Assays, Antitumor , Hydrazones/chemistry , Hydrazones/pharmacology , Lung Neoplasms/drug therapy , Pyrazoles/chemistry , Pyrazoles/pharmacology , bcl-2-Associated X Protein/antagonists & inhibitors , Animals , Apoptosis/drug effects , Cell Survival/drug effects , Cells, Cultured , Female , Fluorescent Antibody Technique , Humans , Immunoblotting , Immunoprecipitation , Jurkat Cells , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitochondria/drug effects , Mitochondria/metabolism , Models, Molecular , Structure-Activity Relationship , bcl-2-Associated X Protein/physiologyABSTRACT
A library of 41 aryloxyimino amides was prepared via solution phase parallel synthesis by extending the multicomponent reaction of (Z)-chlorooximes and isocyanides to the use of electron-deficient phenols. The resulting aryloxyiminoamide derivatives can be used as intermediates for the synthesis of benzo[d]isoxazole-3-carboxamides, dramatically reducing the number of synthetic steps required by other methods reported in literature.
Subject(s)
Amides/chemical synthesis , Combinatorial Chemistry Techniques/methods , Cyanides/chemistry , Oximes/chemistry , Phenols/chemistry , Amides/chemistry , Cyanides/chemical synthesis , Halogenation , Hydrocarbons, Aromatic/chemical synthesis , Hydrocarbons, Aromatic/chemistry , Oximes/chemical synthesis , Phenols/chemical synthesisABSTRACT
(Z)-Arylchlorooximes and α-isocyanoacetamides undergo a smooth reaction to produce 1,3-oxazol-2-oxime derivatives in good yields. Opening of the oxazole ring and deoximation reaction give a facile access to aryl-α-ketoamide amides, a class of privileged scaffolds in medicinal chemistry and important synthetic intermediates in organic chemistry.
