The development of cruzain inhibitors has been driven by the urgent need to develop novel and more effective drugs for the treatment of Chagas' disease. Herein, we report the lead optimization of a class of noncovalent cruzain inhibitors, starting from an inhibitor previously cocrystallized with the enzyme (K i = 0.8 μM). With the goal of achieving a better understanding of the structure−activity relationships, we have synthesized and evaluated a series of over 40 analogues, leading to the development of a very promising competitive inhibitor (8r, IC 50 = 200 nM, K i = 82 nM). Investigation of the in vitro trypanocidal activity and preliminary cytotoxicity revealed the potential of the most potent cruzain inhibitors in guiding further medicinal chemistry efforts to develop drug candidates for Chagas' disease.

Chagas disease causes ~10,000 deaths each year, mainly in Latin America, where it is endemic. The currently available chemotherapeutic agents are ineffective in the chronic stage of the disease, and the lack of pharmaceutical innovation for Chagas disease highlights the urgent need for the development of new drugs. The enzyme cruzain, the main cysteine protease of Trypanosoma cruzi, has been explored as a validated molecular target for drug discovery. Herein, the design, molecular modeling studies, synthesis, and biological evaluation of cyclic imides as cruzain inhibitors are described. Starting with a micromolar-range cruzain inhibitor (3a, IC50 = 2.2 μM), this molecular optimization strategy resulted in the nanomolar-range inhibitor 10j (IC50 = 0.6 μM), which is highly active against T. cruzi intracellular amastigotes (IC50 = 1.0 μM). Moreover, most compounds were selective toward T. cruzi over human fibroblasts, which were used as host cells, and are less toxic to hepatic cells than the marketed drug benznidazole. This study enabled the discovery of novel chemical diversity and established robust structure-activity relationships to guide the design of optimized cruzain inhibitors as new trypanocidal agents.

A virtual screening conducted with nearly 4 000 000 compounds from lead-like and fragment-like subsets enabled the identification of a small-molecule inhibitor (1) of the Trypanosoma cruzi cruzain enzyme, a validated drug target for Chagas disease. Subsequent comprehensive structure-based drug design and structure−activity relationship studies led to the discovery of carbamoyl imidazoles as potent, reversible, and competitive cruzain inhibitors. The most potent carbamoyl imidazole inhibitor (45) exhibited high affinity with a K i value of 20 nM, presenting both in vitro and in vivo activity against T. cruzi. Furthermore, the most promising compounds reduced parasite burden in vivo and showed no toxicity at a dose of 100 mg/kg. These carbamoyl imidazoles are structurally attractive, nonpeptidic, and easy to prepare and synthetically modify. Finally, these results further advance our understanding of the noncovalent mode of inhibition of this pharmaceutically relevant enzyme, building strong foundations for drug discovery efforts.

Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is an enticing antimalarial drug target. Novel chemotypes are needed because existing inhibitors have safety issues that may prevent further development. This work demonstrates isoxazole-based compounds are potent ATP competitive inhibitors of PfPKG and discloses a new analogue in this series. Isoxazoles 3 and 5 had K i values that are comparable to a known standard, 4-[2-(4-fluorophenyl)-5-(1-methylpiperidine-4-yl)-1H pyrrol-3-yl] pyridine. They also exhibited excellent selectivity for PfPKG over the human orthologue and the gatekeeper mutant T618Q PfPKG, which mimics the less accessible binding site of the human orthologue. The human orthologue's larger binding site volume is predicted to explain the selectivity of the inhibitors for the P. falciparum enzyme.

Resumo -O objetivo deste trabalho foi verificar o efeito da época de poda e de sistemas de condução sobre crescimento, produção e qualidade de frutos da figueira cultivada na região norte mineira, sob irrigação. O delineamento utilizado foi em blocos ao acaso, com quatro repetições, e os tratamentos arranjados em esquema fatorial 4x2, tomando por fatores épocas de poda (junho, setembro, dezembro e março) e sistemas de condução (com desponte e sem desponte). Em cada parcela, constituída de três plantas úteis, foram coletados dados nos ciclos de produção 2000/2001 e 2001/2002. Não houve interação entre os fatores época de poda e sistema de condução. Houve diferenças de produção entre épocas de poda; plantas podadas em março e junho apresentaram maiores produções. Não houve diferença de produção entre os dois sistemas de condução. Diferentes épocas de poda e sistemas de condução não influíram na qualidade dos frutos em relação a teores de sólidos solúveis totais, pH, acidez total titulável, açúcares totais, glucose, sacarose e amido. A poda da figueira em diferentes épocas do ano pode propiciar produção na entressafra da cultura no norte de Minas Gerais, sem alterar a qualidade dos frutos verdes com padrão para a indústria.Termos para indexação: Ficus carica, crescimento de frutos, qualidade de frutos. Pruning and conduction system in the yield of green figAbstract -The objective of the work was to verify the effect of pruning time and conduction system on growth, yield, and quality of the fig tree fruit grown in the northern Minas Gerais region, under irrigation. The design utilized was randomized blocks with four replicates, and the treatments arranged in a 4x2 factorial scheme, taking as factors pruning times (June, September, December and March) and conduction system (with lopping and without lopping). In each plot, made up of three useful plants, data concerning the cropping cycles of 2000/2001 and 2001/2002 were collected. No interaction between the factors pruning time and conduction system was verified. There were yield differences between pruning times; the March-and June-pruned plants presented higher yields. There were no yield differences between the conduction systems. Different pruning times and conduction systems did not affect quality of fig tree fruit and contents of total soluble solids, pH, total titrable acidity, total sugars, glucose, sucrose and starch.