ABSTRACT
Affinity selection screening of macrocycle libraries derived from DNA-programmed chemistry identified XIAP BIR2 and BIR3 domain inhibitors that displace bound pro-apoptotic caspases. X-ray cocrystal structures of key compounds with XIAP BIR2 suggested potency-enhancing structural modifications. Optimization of dimeric macrocycles with similar affinity for both domains were potent pro-apoptotic agents in cancer cell lines and efficacious in shrinking tumors in a mouse xenograft model.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/therapeutic use , Breast Neoplasms/drug therapy , Macrocyclic Compounds/chemistry , Macrocyclic Compounds/therapeutic use , X-Linked Inhibitor of Apoptosis Protein/antagonists & inhibitors , Animals , Antineoplastic Agents/pharmacokinetics , Breast/drug effects , Breast/metabolism , Breast/pathology , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Caspase 3/metabolism , Cell Line, Tumor , Crystallography, X-Ray , Drug Discovery , Female , Gene Library , Humans , Macrocyclic Compounds/pharmacokinetics , Mice , Models, Molecular , X-Linked Inhibitor of Apoptosis Protein/metabolismABSTRACT
Since the cloning of Aequorea victoria green fluorescent protein (GFP) in 1992, a family of known GFP-like proteins has been growing rapidly. Today, it includes more than a hundred proteins with different spectral characteristics cloned from Cnidaria species. For some of these proteins, crystal structures have been solved, showing diversity in chromophore modifications and conformational states. However, we are still far from a complete understanding of the origin, functions and evolution of the GFP family. Novel proteins of the family were recently cloned from evolutionarily distant marine Copepoda species, phylum Arthropoda, demonstrating an extremely rapid generation of fluorescent signal. Here, we have generated a non-aggregating mutant of Copepoda fluorescent protein and solved its high-resolution crystal structure. It was found that the protein beta-barrel contains a pore, leading to the chromophore. Using site-directed mutagenesis, we showed that this feature is critical for the fast maturation of the chromophore.
Subject(s)
Arthropods/chemistry , Copepoda/chemistry , Green Fluorescent Proteins/chemistry , Green Fluorescent Proteins/metabolism , Protein Biosynthesis , Animals , Arthropods/metabolism , Copepoda/metabolism , Crystallography, X-Ray/methods , Embryo, Nonmammalian , Green Fluorescent Proteins/genetics , HeLa Cells , Humans , Models, Molecular , Protein Conformation , Protein Folding , Protein Structure, Tertiary , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Structural Homology, Protein , Xenopus laevis/embryologyABSTRACT
The sulfamic acid phosphotyrosine mimetic was coupled with a previously known malonate template to obtain highly selective and potent inhibitors of HPTPbeta. Potentially hydrolyzable malonate ester functionalities were replaced with 1,2,4-oxadiazoles without a significant effect on HPTPbeta potency.
Subject(s)
Chemistry, Pharmaceutical/methods , Nerve Tissue Proteins/antagonists & inhibitors , Protein Tyrosine Phosphatases/antagonists & inhibitors , Crystallography, X-Ray , Drug Design , Hydrogen Bonding , Hydrolysis , Models, Chemical , Models, Molecular , Molecular Structure , Receptor-Like Protein Tyrosine Phosphatases, Class 5 , Structure-Activity RelationshipABSTRACT
High-throughput screening of the P&GP corporate repository against several protein tyrosine phosphatases identified the sulfamic acid moiety as potential phosphotyrosine mimetic. Incorporation of the sulfamic acid onto a 1,2,3,4-tetrahydroisoquinoline scaffold provided a promising starting point for PTP1B inhibitor design.
