ABSTRACT
Respiratory syncytial virus (RSV) represents a threat to infants, the elderly, and the immunocompromised. RSV entry blockers are in clinical trials, but escape mutations challenge their potential. In search of RSV inhibitors, we have integrated a signature resistance mutation into a recombinant RSV virus and applied the strain to high-throughput screening. Counterscreening of candidates returned 14 confirmed hits with activities in the nano- to low-micromolar range. All blocked RSV polymerase activity in minigenome assays. Compound 1a (GRP-74915) was selected for development based on activity (EC50 = 0.21 µM, selectivity index (SI) 40) and scaffold. Resynthesis confirmed the potency of the compound, which suppressed viral RNA synthesis in infected cells. However, metabolic testing revealed a short half-life in the presence of mouse hepatocyte fractions. Metabolite tracking and chemical elaboration combined with 3D-quantitative structure-activity relationship modeling yielded analogues (i.e., 8n: EC50 = 0.06 µM, SI 500) that establish a platform for the development of a therapeutic candidate.
Subject(s)
Antiviral Agents/chemistry , Antiviral Agents/pharmacology , DNA-Directed RNA Polymerases/antagonists & inhibitors , Drug Design , Respiratory Syncytial Virus Infections/drug therapy , Respiratory Syncytial Virus, Human/drug effects , Respiratory Syncytial Virus, Human/enzymology , Animals , Antiviral Agents/metabolism , Cell Line , DNA-Directed RNA Polymerases/metabolism , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Humans , Mice , Quantitative Structure-Activity Relationship , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Human/metabolism , Small Molecule Libraries/chemistry , Small Molecule Libraries/metabolism , Small Molecule Libraries/pharmacology , Viral Proteins/antagonists & inhibitors , Viral Proteins/metabolismABSTRACT
A substrate-induced stereocontrol strategy was used to gain access to the tetracyclic core of (-)-lemonomycin. An advanced intermediate was prepared from a known substituted tyrosinol through a 16-step sequence, which involved a Pictet-Spengler reaction, a [3+2] dipolar cycloaddition and an enamide hydrogenation.
ABSTRACT
Natural concentration of antimalaric components in Tropical arthropods (in vitro). Alcohol, hexane and dichlorometane extracts of 751 samples of Costa Rican arthropods were studied for the presence of antimalaric components. With Plasmodium berghei we set an in vitro model in which the effect of the extract was determined by staining of the parasites with cresil brilliant blue. Active extracts at concentration of 50 mg or less, were considered positive. Promissory extracts were found in the orders Lepidoptera (24.1%), Coleoptera (32.8%), Hemiptera (38.5%) and Polydesmida (81.3%). Since most of the Lepidoptera samples were in the immature stages, the relation with the host plant was analyzed. Cannaceae, Flacourtiaceae, Crisobalanaceae, Lauraceae, Fagaceae, Ulmaceae, Rosaceae, Asteraceae, Rubiaceae, Lauraceae and Caprifoliaceae were related with the Lepidoptera larvae, and an antimalaric effect has been reported in most of these families. In the orders Polydesmida, Opiliones and Blattodea, the extract from adults also had some important effect, probably because all of them fed on plants. Polydesmida and Opiliones have chemical substances that probably serve as defensive purposes; these chemicals could also have some antiparasitic effect. Therefore, the detection of antimalaric components in arthropod species led to the identification of plants with promissory antimalaric components. Rev. Biol. Trop. 56 (2): 473-485. Epub 2008 June 30.
Extractos alcohólicos, hexánicos y diclorometánicos de 751 muestras de artrópodos fueron estudiados por la presencia de actividad antimalárica. En este trabajo se empleó un modelo murino usando el Plasmodium berghei, modelo que es biológicamente similar a la malaria humana. El estudio fue realizado determinando el efecto del extracto sobre el parásito por la inclusión o no del colorante azul de cresil brillante. Estimando como positivos aquellos extractos cuya actividad antimalárica se mostró en concentraciones no mayores de 50 mg, se encontró que los órdenes más promisorios fueron Lepidoptera (24.1%), Polydesmida (81.3%), Blattodea (25%) y Opiliones, entre otros. Las formas inmaduras de Lepidoptera fueron las más positivas, por lo que se analizaron las plantas hospederos de donde se alimentaban dichos organismos. Las familias de estas plantas eran Malvaceae, Acanthaceae, Rutaceae, Myrtaceae, Solanaceae, Fabaceae, Urticaceae, Anacardiaceae, Rosaceae, Asteraceae, Rubiaceae, Lauraceae y Caprifoliaceae. Especies de casi todas estas familias han sido reportadas con actividad antimalárica. En el caso de los órdenes Polydesmida, Opiliones y Blattodea, cuyas formas adultas presentaron alguna actividad contra P. berghei, encontramos que todos esos grupos se alimentan también de plantas. En el caso de Opiliones sus especies son predadores de lepidópteros, coleópteros, hemípteros fitófagos y otros artrópodos, además de que producen sustancias de defensas tales como alcoholes, cetonas y quinonas, entre otros, todo lo cual podría explicar la actividad encontrada. Algunas especies del Orden Polydesmida, también secretan ciertas sustancias químicas, las cuales podrían tener un efecto antiparasitario. Así, a través de este trabajo en artrópodos hemos llegado a identificar fuentes vegetales potenciales para componentes antimaláricos.
Subject(s)
Animals , Mice , Antimalarials/pharmacology , Arthropods/chemistry , Feeding Behavior/physiology , Malaria/drug therapy , Plant Extracts/pharmacology , Tissue Extracts/pharmacology , Antimalarials/isolation & purification , Arthropods/classification , Arthropods/physiology , Disease Models, Animal , Tissue Extracts/isolation & purificationABSTRACT
Alcohol, hexane and dichlorometane extracts of 751 samples of Costa Rican arthropods were studied for the presence of antimalaric components. With Plasmodium berghei we set an in vitro model in which the effect of the extract was determined by staining of the parasites with cresil brilliant blue. Active extracts at concentration of 50 mg or less, were considered positive. Promissory extracts were found in the orders Lepidoptera (24.1%), Coleoptera (32.8%), Hemiptera (38.5%) and Polydesmida (81.3%). Since most of the Lepidoptera samples were in the immature stages, the relation with the host plant was analyzed. Cannaceae, Flacourtiaceae, Crisobalanaceae, Lauraceae, Fagaceae, Ulmaceae, Rosaceae, Asteraceae, Rubiaceae, Lauraceae and Caprifoliaceae were related with the Lepidoptera larvae, and an antimalaric effect has been reported in most of these families. In the orders Polydesmida, Opiliones and Blattodea, the extract from adults also had some important effect, probably because all of them fed on plants. Polydesmida and Opiliones have chemical substances that probably serve as defensive purposes; these chemicals could also have some antiparasitic effect. Therefore, the detection of antimalaric components in arthropod species led to the identification of plants with promissory antimalaric components.
Subject(s)
Antimalarials/pharmacology , Arthropods/chemistry , Feeding Behavior/physiology , Malaria/drug therapy , Plant Extracts/pharmacology , Tissue Extracts/pharmacology , Animals , Antimalarials/isolation & purification , Arthropods/classification , Arthropods/physiology , Disease Models, Animal , Mice , Tissue Extracts/isolation & purificationABSTRACT
Treatment of toxoplasmosis usually causes secondary effects. It is important to find active substances extracted from natural organisms. In this work we studied some arthropod extracts that have effect against Toxoplasma multiplication inside mouse macrophages. After studying 382 extracts, 23 were selected on the basis of the activity and we found that 13 extracts from orders Polydesmida, Lepidoptera, Orthoptera and Hymenoptera exerted an important inhibition of Toxoplasma multiplication.
Subject(s)
Animals , Mice , Arthropods , Macrophages, Peritoneal , Tissue Extracts , Toxoplasma , Arthropods , Time FactorsABSTRACT
Treatment of toxoplasmosis usually causes secondary effects. It is important to find active substances extracted from natural organisms. In this work we studied some arthropod extracts that have effect against Toxoplasma multiplication inside mouse macrophages. After studying 382 extracts, 23 were selected on the basis of the activity and we found that 13 extracts from orders Polydesmida, Lepidoptera, Orthoptera and Hymenoptera exerted an important inhibition of Toxoplasma multiplication.
