Efficacy of 2-hydroxy-3-phenylsulfanylmethyl-[1,4]-naphthoquinone derivatives against different Trypanosoma cruzi discrete type units: Identification of a promising hit compound.

The limited efficacy of benznidazole (Bz) indicated by failures of current Phase II clinical trials emphasizes the urgent need to identify new drugs with improved safety and efficacy for treatment of Chagas disease (CD). Herein, we analyzed the efficacy of a series of 2-hydroxy-3-phenylsulfanylmethyl-[1,4]-naphthoquinones against different Trypanosoma cruzi discrete type units (DTUs) of relevant clinical forms of CD. Cytotoxic and trypanocidal effect of naphthoquinone derivatives were assessed in mammalian cells, trypomastigotes and intracellular amastigotes using, luminescent assays (CellTiter-Glo and T. cruzi Dm28c-luciferase) and/or counting with a light microscope. Reactive oxygen species (ROS) production and intracellular targets of promising compounds were assessed with 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) probe and ultrastructural analysis, respectively. ADMET properties were analyzed by in silico modeling. Most of the compounds showed low cytotoxic effect. Only two compounds (Compounds 2 and 11) had IC50 values lower than Bz, showing higher susceptibility of bloodstream trypomastigotes. Compound 2 exhibited greater efficacy against trypomastigotes from different T. cruzi DTUs, even better than Bz against Brazil and CL strains. Ultrastructural analysis revealed changes in intracellular compartments, suggesting autophagy as one possible mechanism of action. Oxidative stress, induced by Compound 2, resulted in elevated level of ROS, leading to parasite death. Compound 2 was also effective against intracellular amastigotes, showing high selectivity index. ADMET analysis predicted good oral bioavailability, reduced drug metabolism and no carcinogenic potential for Compound 2. The data highlight Compound 2 as a hit compound and stimulate further structural and pharmacological optimization to potentiate its trypanocidal activity and selectivity.

Searching for a potential antibacterial lead structure against bacterial biofilms among new naphthoquinone compounds.

AIMS: The aims of this study were to design, synthesize and to evaluate 2-hydroxy-3-phenylsulfanylmethyl-[1,4]-naphthoquinones against Gram-negative and Gram-positive bacterial strains, including methicillin-resistant Staphylococcus aureus (MRSA) and its biofilm, to probe for potential lead structures. METHODS AND RESULTS: Thirty-six new analogues were prepared with good yields using a simple, fast, operational three-procedure reaction and a thiol addition to an ο-quinone methide using microwave irradiation. All compounds were tested against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Proteus mirabilis ATCC 15290, Serratia marcescens ATCC 14756, Klebsiella pneumoniae ATCC 4352, Enterobacter cloacae ATCC 23355, Enterococcus faecalis ATCC 29212, S. aureus ATCC 25923, Staphylococcus simulans ATCC 27851, Staphylococcus epidermidis ATCC 12228 and a hospital strain of MRSA. Their antibacterial activity was determined using the disc diffusion method, revealing the activity of 19 compounds, mainly against Gram-positive strains. Interestingly, the minimal inhibitory concentration ranges detected for the hit molecules (32-128 µg ml-1 ) were within Clinical and Laboratory Standards Institute levels. Promisingly, compound 15 affected the MRSA strain, with a reduction of up to 50% in biofilm formation, which is better than vancomycin as biofilm forms a barrier against the antibiotic that avoids its action. CONCLUSIONS: After probing 36 naphthoquinones for a potential antibacterial lead structure against the bacterial biofilm, we found that compound 15 should be explored further and also should be structurally modified in the near future to test against Gram-negative strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Since vancomycin is one of the last treatment options currently available, and it is unable to inhibit biofilm, the research of new antimicrobials is urgent. In this context, 2-hydroxy-3-phenylsulfanylmethyl-[1,4]-naphthoquinones proved to be a promising lead structure against MRSA and bacterial biofilm.

Variação de matéria seca e de nutrientes nas folhas e nos frutos, produção de ácido ascórbico e suco, em seis cultivares de citros, durante um ciclo

The experiment was carried out in a commercial citrus orchard located in Mogi-Guaçu (22° 22'S., 46° 56'WGr.), State of São Paulo, Brazil. Five orange types were studied: Cravo (Citrus reticulata Blanco); Hamlin (Citrus sinensis (L.) Osbeck); Natal (Citrus sinensis (L.) Osbeck); Valencia (Citrus sinensis (L.) Osbeck) and Pera (Citrus sinensis (L.) Osbeck) and a tangerine Murcott (Citrus reticulata Blanco x Citrus sinensis (L.) Osbeck). Fruits were picked from 30 days after flowering to the peak of commercial harvest. Leaves nearest the fruits were collected at the same time as the fruits. Sampled material was used to determine: fruit dry weight, macronutrient and micronutrient concentration in the leaves, nutrient extraction by the fruits, juice production per fruit and ascorbic acid concentration in the juice. Fruit dry weight increased following the second month after flowering. Except for Cravo orange fruit dry weight decreased at the end of the cycle. Macronutrient and micronutrient concentrations in the leaves showed variations during fruit development. Rate of nutrient extraction in decreasing order was: K, N, Ca, Mg, P = S, Fe, B, Zn, Mn and Cu. The cultivar that exported the largest amount of nutrients was Pera followed by Hamlin = Cravo, Murcott, Valencia and Natal. Juice per fruit ranged between 43 and 95 mililiters whereas ascorbic acid concentration ranged between 30 and 95 miligrams per 100 mililiters of juice.

De uma plantação de citros, com os cultivares T. Cravo (Citrus reticulata Blanco), L.Hamlin (Citrus sinensis (L.) Osbeck), T. Murcott (Citrus reticulata Blanco x Citrus sinensis (L.) Osbeck), L. Natal (Citrus sinensis (L.) Osbeck, L. Valencia (Citrus sinensis (L.) Osbeck) e L. Pera (Citrus sinensis (L.) Osbeck), situada na "Fazenda Sete Lagoas", no município de Mogi-Guaçu (22° 22% 46° 56'W.Gr.), em Latossolo Vermelho amarelo, fase arenosa, foram coletados frutos 30 dias após florescimento, até a idade da coleta comercial. No material coletado, foram determinadas a variação da matéria seca, a concentração dos macro e micronutrientes nas folhas adjacentes ao fruto, a extração de macro e micronutríentes pelos frutos, a produção de suco (ml) por fruto e a concentração de ácido ascórbico (mg/100 ml de suco). Concluiu-se que: 1. O aumento da matéria seca, intensifica-se a partir do segundo mês apos o florescimento; 2. Com exceção da T. Cravo, ocorre uma diminuição na produção de matéria seca no final do ciclo; 3. A concentração dos macro e micronutrientes nas folhas apresenta oscilações durante o desenvolvimento do fruto; 4. A ordem decrescente de extração de nutrientes é: K, N, Ca, Mg, P = S, Fe, B, Zn, Mn, Cu; 5. A capacidade de exportação de nutrientes pelos cultivares é, em ordem decrescente: L. Pera, L. Hamlin = T. Cravo, T. Murcott, L. Valencia, L. Natal; 6. A quantidade de suco produzido por fruto, oscila entre 43 a 95 ml; 7. A concentração de ácido ascórbico (mg/100 ml de suco), varia entre 30 a 95.

Variação de matéria seca e de nutrientes nas folhas e nos frutos, produção de ácido ascórbico e suco, em seis cultivares de citros, durante um ciclo

The experiment was carried out in a commercial citrus orchard located in Mogi-Guaçu (22° 22'S., 46° 56'WGr.), State of São Paulo, Brazil. Five orange types were studied: Cravo (Citrus reticulata Blanco); Hamlin (Citrus sinensis (L.) Osbeck); Natal (Citrus sinensis (L.) Osbeck); Valencia (Citrus sinensis (L.) Osbeck) and Pera (Citrus sinensis (L.) Osbeck) and a tangerine Murcott (Citrus reticulata Blanco x Citrus sinensis (L.) Osbeck). Fruits were picked from 30 days after flowering to the peak of commercial harvest. Leaves nearest the fruits were collected at the same time as the fruits. Sampled material was used to determine: fruit dry weight, macronutrient and micronutrient concentration in the leaves, nutrient extraction by the fruits, juice production per fruit and ascorbic acid concentration in the juice. Fruit dry weight increased following the second month after flowering. Except for Cravo orange fruit dry weight decreased at the end of the cycle. Macronutrient and micronutrient concentrations in the leaves showed variations during fruit development. Rate of nutrient extraction in decreasing order was: K, N, Ca, Mg, P = S, Fe, B, Zn, Mn and Cu. The cultivar that exported the largest amount of nutrients was Pera followed by Hamlin = Cravo, Murcott, Valencia and Natal. Juice per fruit ranged between 43 and 95 mililiters whereas ascorbic acid concentration ranged between 30 and 95 miligrams per 100 mililiters of juice.

De uma plantação de citros, com os cultivares T. Cravo (Citrus reticulata Blanco), L.Hamlin (Citrus sinensis (L.) Osbeck), T. Murcott (Citrus reticulata Blanco x Citrus sinensis (L.) Osbeck), L. Natal (Citrus sinensis (L.) Osbeck, L. Valencia (Citrus sinensis (L.) Osbeck) e L. Pera (Citrus sinensis (L.) Osbeck), situada na "Fazenda Sete Lagoas", no município de Mogi-Guaçu (22° 22% 46° 56'W.Gr.), em Latossolo Vermelho amarelo, fase arenosa, foram coletados frutos 30 dias após florescimento, até a idade da coleta comercial. No material coletado, foram determinadas a variação da matéria seca, a concentração dos macro e micronutrientes nas folhas adjacentes ao fruto, a extração de macro e micronutríentes pelos frutos, a produção de suco (ml) por fruto e a concentração de ácido ascórbico (mg/100 ml de suco). Concluiu-se que: 1. O aumento da matéria seca, intensifica-se a partir do segundo mês apos o florescimento; 2. Com exceção da T. Cravo, ocorre uma diminuição na produção de matéria seca no final do ciclo; 3. A concentração dos macro e micronutrientes nas folhas apresenta oscilações durante o desenvolvimento do fruto; 4. A ordem decrescente de extração de nutrientes é: K, N, Ca, Mg, P = S, Fe, B, Zn, Mn, Cu; 5. A capacidade de exportação de nutrientes pelos cultivares é, em ordem decrescente: L. Pera, L. Hamlin = T. Cravo, T. Murcott, L. Valencia, L. Natal; 6. A quantidade de suco produzido por fruto, oscila entre 43 a 95 ml; 7. A concentração de ácido ascórbico (mg/100 ml de suco), varia entre 30 a 95.