RESUMO
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.
Assuntos
Antineoplásicos/química , Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Compostos Macrocíclicos/química , Compostos Macrocíclicos/uso terapêutico , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/antagonistas & inibidores , Animais , Antineoplásicos/farmacocinética , Mama/efeitos dos fármacos , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Cristalografia por Raios X , Descoberta de Drogas , Feminino , Biblioteca Gênica , Humanos , Compostos Macrocíclicos/farmacocinética , Camundongos , Modelos Moleculares , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismoRESUMO
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.
Assuntos
Artrópodes/química , Copépodes/química , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Biossíntese de Proteínas , Animais , Artrópodes/metabolismo , Copépodes/metabolismo , Cristalografia por Raios X/métodos , Embrião não Mamífero , Proteínas de Fluorescência Verde/genética , Células HeLa , Humanos , Modelos Moleculares , Conformação Proteica , Dobramento de Proteína , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Homologia Estrutural de Proteína , Xenopus laevis/embriologiaRESUMO
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.
Assuntos
Química Farmacêutica/métodos , Proteínas do Tecido Nervoso/antagonistas & inibidores , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Cristalografia por Raios X , Desenho de Fármacos , Ligação de Hidrogênio , Hidrólise , Modelos Químicos , Modelos Moleculares , Estrutura Molecular , Proteínas Tirosina Fosfatases Classe 5 Semelhantes a Receptores , Relação Estrutura-AtividadeRESUMO
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.
