In this report, we designed and synthesized ten N-(R-phenyl)-3-carboxamide-coumarin derivatives (2a-2j), exploring the coumarin nucleus, and an R-phenyl group as a structural scaffold and a peptide bond as a linker between them. The structure-activity relationships were investigated with different R-substituents (H, Br, NO2, Cl) in ortho, meta, or para positions from the phenyl group. Coumarins were obtained in good yields (72-95%), and in vitro screening against Canavalia ensiformis urease showed potential inhibitory percentages ranging from 42 to 65%. Half-maximal inhibitory concentration (IC50) values were determined for the best compounds 2b (R = 2-Br) and 2d (R = 4-Br). After conducting molecular docking and molecular dynamics on urease from C. ensiformis and Helicobacter pylori, potential binding modes for the most effective compounds 2b and 2d showed that these derivatives are able to interact with the crucial residue Cys592, thereby blocking the access of the urea substrate to the active site. According to density functional theory (DFT) calculations, 4-Br substitution on H. pylori urease was necessary for interacting with catalytic amino acids on the active site. report NRphenyl3carboxamidecoumarin NRphenylcarboxamidecoumarin N R 3 carboxamide 2a2j, 2a2j aj 2a 2j , j (2a-2j) nucleus Rphenyl them structureactivity structure activity Rsubstituents substituents H, H (H Br NO2 NO Cl ortho meta 7295%, 7295 72 95% 95 (72-95%) 4 65 65% Halfmaximal Half maximal IC50 IC (IC50 b 2Br 2 2-Br d 4Br. 4Br . 4-Br) C Cys592 Cys site DFT (DFT calculations NRphenyl carboxamidecoumarin (2a-2j 7295% 729 7 9 (72-95% 6 IC5 (IC5 Cys59 (72-95 (IC Cys5 (72-9 (72- (72 (7 (

Abstract Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are the latest class of drugs approved to treat type 2 DM (T2DM). Although adverse effects are often caused by a metabolite rather than the drug itself, only the safety assessment of disproportionate drug metabolites is usually performed, which is of particular concern for drugs of chronic use, such as SGLT2i. Bearing this in mind, in silico tools are efficient strategies to reveal the risk assessment of metabolites, being endorsed by many regulatory agencies. Thereby, the goal of this study was to apply in silico methods to provide the metabolites toxicity assessment of the SGLT2i. Toxicological assessment from SGLT2i metabolites retrieved from the literature was estimated using the structure and/or statistical-based alert implemented in DataWarrior and ADMET predictorTM softwares. The drugs and their metabolites displayed no mutagenic, tumorigenic or cardiotoxic risks. Still, M1-2 and M3-1 were recognized as potential hepatotoxic compounds and M1-2, M1-3, M3-1, M3-2, M3-3 and M4-3, were estimated to have very toxic LD50 values in rats. All SGLT2i and the metabolites M3-4, M4-1 and M4-2, were predicted to have reproductive toxicity. These results support the awareness that metabolites may be potential mediators of drug-induced toxicities of the therapeutic agents.

Abstract The concern about the risks of viral infections transmission through blood transfusion has led into a search for improvements on screening tests used for the selection of blood donors. Molecular biology techniques applied in researches of viral genomes, known as Nucleic Acid-amplification-Test (NAT), represent a technology capable of increasing transfusion safety by shortening the diagnostic window period. In Brazil, the implementation of this technology for the detection of HIV, HCV and HBV occurred due to the implantation of the NAT Kit – produced by Immunobiological Technology Institute (Biomanguinhos-FIOCRUZ), in the Brazilian blood centers. The National Health Surveillance Agency attaches great importance to validation, since it standardizes, disciplines and regulates criteria for the registration of health products. This work aims to establish a protocol of performance validation by real-time PCR method, taking as the object of study the Bio-Manguinhos NAT Kit, in order to update the product registration or to meet any future needs to ensure all regulatory requirements for the performance validation of the real-time PCR diagnostic kit. The protocol developed followed the ICH recommendations. The results revealed that the adopted methodology contemplates the necessary requirements for compliance with the Brazilian legislation, as well as the established validation parameters.

Abstract Sildenafil is a potent selective inhibitor of phosphosdiesterase-5 previously used in erectile dysfunction and subsequently approved in 2005 for pulmonary arterial hypertension treatment. Since oral administration of sildenafil shows pharmacokinetic problems with mean absolute bioavailability of 41%, the goal of this work was to develop a novel sildenafil self-emulsifying drug delivery system (SEDDS) for oral absorption improvement and management of dosage. One pharmaceutical solution and four SEDDS containing sildenafil were successfully obtained and SEDDS formed O/W nanoemulsion with droplet size less than 300 nm. The stability studies evidenced that the SEDDS containing 3.3% w/w of sildenafil yielded the best results. The safety of 2-pyrrolidone/isobutanol in oral formulations was assessed in mice and no lethality was achieved in the placebo groups with LD50 of 490 mg/Kg for SEDDS II-3.3, suggesting it as a safe excipient for humans. Therewithal, in silico studies using PBPK models provided the pharmacokinetic profile of sildenafil SEDDS. Subsequently, in silico evaluation indicated that the sildenafil SEDDS droplet size influenced its bioavailability, enhancing absorption, assuring a good pharmacokinetic profile. These findings suggest that the formulations developed here presented the potential to enhance drug oral absorption with the possibility to control drug dosage as they are liquid pharmaceutical formulations.

This article reports a novel virtual screening algorithm seeking the rational identification of novel lead anticoagulants. Seven 5-(3-methyl-1-aryl-1H-pyrazol-4-yl)-1H-tetrazoles and seven novel 1-aryl-4-(4,5-dihydro-1H-imidazol-2-yl)-3-methyl-1H-pyrazoles were obtained in three steps starting from arylhydrazine hydrochlorides as raw materials in good yields: 50-72% and 50-85%, respectively. All compounds were submitted to an in silico target-base pipeline named integrated multiplex analysis virtual screening (IMA-VS), which comprises the evaluation of their (i) fitting physicochemical properties to the chemical environment of the target enzyme; (ii) active-site homing electrostatic potential to the target enzyme; (iii) structural fitting to the target active site through molecular docking; and (iv) overall absorption, distribution, metabolism, excretion and toxicity (ADMET) profile. After the virtual selection of potential anticoagulant hits, all molecules were synthesized and candidates were evaluated in vitro for their anticoagulant and hemolytic profile. The most promising candidate pointed out by IMA-VS was compound 1-(3',4'-dichlorophenyl)-4-(4,5-dihydro-1H-imidazol-2-yl)-3-methyl-1H-pyrazole that shown to display factor Xa (FXa)-specific inhibitory activity in vitro, acting as an uncompetitive inhibitor with an inhibition constant (Ki) = 61.16 ± 12.96 µM, in addition to the lowest hemolytic activity of the series. Further experiments revealed the antithrombotic activity of this compound in an in vivo model of arterial thrombosis induced by FeCl3.

ABSTRACT This study presents the bioreduction of six β-ketoesters by whole cells of Kluyveromyces marxianus and molecular investigation of a series of 13 β-ketoesters by hologram quantitative structure-activity relationship (HQSAR) in order to relate with conversion and enantiomeric excess of β-stereogenic-hydroxyesters obtained by the same methodology. Four of these were obtained as (R)-configuration and two (S)-configuration, among them four compounds exhibited >99% enantiomeric excess. The β-ketoesters series LUMO maps showed that the β-carbon of the ketoester scaffold are exposed to undergo nucleophilic attack, suggesting a more favorable β-carbon side to enzymatic reduction based on adopted molecular conformation at the reaction moment. The HQSAR method was performed on the β-ketoesters derivatives separating them into those provided predominantly (R)- or (S)-β-hydroxyesters. The HQSAR models for both (R)- and (S)-configuration showed high predictive capacity. The HQSAR contribution maps suggest the importance of β-ketoesters scaffold as well as the substituents attached therein to asymmetric reduction, showing a possible influence of the ester group carbonyl position on the molecular conformation in the enzyme catalytic site, exposing a β-carbon side to the bioconversion to (S)- and (R)-enantiomers.

A melissa (Melissa officinalis) é uma planta medicinal comumente usada como calmante e ingerida na forma de chá. Para otimizar sua produção, este trabalho objetivou avaliar o efeito de diferentes doses de esterco bovino, com relação ao fertilizante mineral na produção de biomassa foliar e teor de óleo essencial. O experimento foi conduzido na Universidade Federal de Uberlândia e no Instituto Agronômico de Campinas. O delineamento estatístico foi de blocos casualisados com seis tratamentos (0, 1, 2, 4, 8 kg m-2 de esterco bovino e 30 g m-2 de NPK 4-14-8), em quatro repetições. O óleo essencial foi extraído por hidrodestilação em equipamento tipo Clevenger modificado. As doses de esterco bovino influenciaram a altura de plantas, massa fresca total e massa seca foliar por planta e por hectare. As duas formas de adubação foram superiores à testemunha para praticamente todas as variáveis, exceto em relação ao comprimento e largura foliar, teor de óleo na matéria seca e fresca foliar. Conclui-se que a melissa responde à adubação orgânica com esterco bovino e adubação mineral para produção de biomassa.

Lemon balm (Melissa officinalis) is a medicinal plant commonly used as a sedative and ingested as a tea. Studies on agricultural practices are required to optimize its yield. To optimize its production, this study evaluated the effect of different doses of organic fertilizer (cow manure) in comparison to mineral fertilizer on biomass production and essential oil yield. The experiment was carried out at the Federal University of Uberlândia, Minas Gerais state, Brazil, and at the Agronomic Institute of Campinas, São Paulo state, Brazil. The experimental design was randomized blocks with six treatments (0, 1, 2, 4, 8 kg m-2 of manure and 30 g m-2 of NPK 4-14-8), and four replications. Hydro-distillation was done with a modified Clevenger distiller for essential oil extraction. Cattle manure influenced plant height, total fresh and dry mass of leaves, per plant and per hectare. The two fertilization forms were better than the non-fertilized control for all variables, except for leaf length and width and oil content in the leaf fresh and dry matter. It can be concluded that lemon balm is responsive to organic fertilization with manure and mineral fertilization for biomass production.

HIV-1 reverse transcriptase (HIV-1 RT) is a therapeutic target for the treatment of HIV-positive individuals or those already showing AIDS symptoms. In this perspective, the identification of new inhibitors for this enzyme is of great importance in view of the growing viral resistance to the existing treatments. This resistance has compromised the quality of life of those infected with multidrug-resistant strains, whose treatment options are already limited, putting at risk these individuals lives. The literature has recognized marine organisms and their products as natural sources for the identification of new therapeutic options for different pathologies. In this brief review, we consider the structure of HIV-1 RT and its most common inhibitors, as well as some marine diterpenes originally reported as HIV-1 RT inhibitors to encourage the identification and development of new marine antiviral prototypes.

Melissa officinalis L., Lamiaceae, is an herb with great growth prospects in the cosmetic industry due to its essential oil. In order to improve its production, it is necessary to study related agricultural practices. This study evaluated the effect of organic and mineral fertilization on the chemical composition of lemon balm (Melissa officinalis L.) essential oil. The assay was conducted at the "Fazenda Experimental do Glória" of the Federal University of Uberlândia, and essential oil extraction and GC/MS analyses were completed by the Centre for Research and Development on Plant Genetic Resources of the Campinas Agronomic Institute. The assay was conducted in a randomized complete block design with three replications. The tested treatments were six types of fertilization (0, 1, 2, 4, 8 kg.m-2 of cattle manure and mineral fertilizing with 60 g.m-2 of NPK 4-14-8 + 4 g.m-2 of boric acid) with four replications. The essential oil was extracted by hydrodistillation in a modified Clevenger apparatus. The chemical composition was analyzed by GC/MS. The essential oil presented the same compounds for all treatments; however, the relative proportion of some chemical constituents was altered according to the treatment. Neral, geranial, and citronellal were the major constituents.

A quimioterapia para as leishmanioses, doenças causadas por protozoários do gênero Leishmania, ainda permanece ineficiente em diversos tratamentos. Portanto, existe a necessidade de pesquisa por novos fármacos. Nesse trabalho, foram sintetizados derivados 1-aril-1H-pirazol-4-carboximidamidas, avaliadas as atividades leishmanicida e os efeitos citotóxicos in vitro, e realizado um estudo de relação estrutura-atividade (REA) com a série de compostos. O composto 2 apresentou um perfil de atividade que pode ser melhorado através de estratégias de modificação molecular da química medicinal.

Chemotherapy for leishmaniasis, diseases caused by protozoa of the genus Leishmania, remains inefficient in several treatments. So there is a need to search for new drugs. In this work, we have synthesized 1-aryl-1H-pyrazole-4-carboximidamides derivatives and evaluated antileishmanial activities in vitro, as well as cytotoxic effects. Structure-activity relationship (SAR) studies were carried out with all the compounds of the series. Compound 2 showed an activity profile that can be improved through medicinal chemistry strategies.

A cenoura é a principal hortaliça em valor econômico cujo produto é a raiz, encontrando-se entre as dez espécies de olerícolas mais cultivadas no Brasil. O presente trabalho avaliou a produtividade, número e qualidade de raízes comerciais de cenoura, cultivar Brasília-RL, em diferentes adubações de cobertura. O experimento foi conduzido num Latossolo Vermelho-Amarelo, da Fazenda Olhos D'Água, em São Gotardo-MG, de janeiro a abril de 2006. O delineamento experimental foi em blocos casualizados com nove tratamentos (T0= testemunha sem cobertura; T1= 100 kg ha-1 de 20-00-20 aos 30 e 45 dias após semeadura; T2= 200 kg ha-1 de 20-00-20 aos 30 e 45 dias após semeadura; T3= 300 kg ha-1 de 20-00-20 aos 30 e 45 dias após semeadura; T4= 200 kg ha-1 de 20-00-20 aos 30 dias após semeadura; T5= 200 kg ha-1 de 20-00-20 aos 45 dias após a semeadura; T6= 300 kg ha-1 de 20-00-20 aos 45 dias após a semeadura; T7= 130 kg ha-1de nitrocálcio aos 30 e 45 dias após semeadura e T8= 130 kg ha-1 de nitrocálcio aos 30 dias após semeadura e T9= 133 kg ha-1 de 15-00-14 aos 45 dias após semeadura) e cinco repetições. Foram avaliados o número e a massa de raízes nas classes comerciais: C-10 (>10<14 cm), C-14 (>14<18 cm), C-18 (>18<22 cm), C- 22 (>22<26 cm) e C-26 (>26 cm), o número e massa de raízes comerciais (C) e não comerciais (NC) e número total de raízes (T). O número total de raízes de cenoura (T) não foi afetado significativamente pelos tratamentos de adubação de cobertura, sendo a máxima produtividade de raízes (27,5 t ha-1), utilizando-se o formulado 20-00-20, obtida com a aplicação de 229,1 kg ha-1. A aplicação conjunta de fontes de nitrogênio, potássio e cálcio aumentou a produtividade de raízes comerciais, principalmente na classe de maior valor (C-18).

The carrot is included among the ten most economically important vegetables in Brazil. We evaluated the root yield, number and quality of commercial carrots, cultivar Brasília-RL, under different top-dressing fertilizations. The experiment was carried out on a Latosol, at Fazenda Olhos D'Água, in São Gotardo, Minas Gerais State, from January to April 2006. The experiment was set up in randomized blocks with nine treatments (T0= no top-dressing control; T1= 100 kg ha-1 20-00-20 at 30 and 45 days after sowing; T2= 200 kg ha-1 20-00-20 at 30 and 45 days after sowing; T3= 300 kg ha-1 20-00-20 at 30 and 45 days after sowing; T4= 200 kg ha-1 20-00-20 at 30 days after sowing; T5= 200 kg ha-1 20-00-20 at 45 days after sowing; T6= 300 kg ha-1 20-00-20 at 45 days after sowing; T7= 130 kg ha-1 nitrocalcium at 30 and 45 days after sowing; T8= 130 kg ha-1 nitrocalcium at 30 days after sowing and T9= 133 kg ha-1 15-00-14 at 45 days after sowing) and five replications. The root number and weight in each commercial class were evaluated: C-10 (>10<14 cm), C-14 (>14<18 cm), C-18 (>18<22 cm), C-22 (>22<26 cm), C-26 (>26 cm), the commercial (C), the non commercial (NC) and total (T) root number and weight were also evaluated. The total number of carrot roots (T) did not present significant differences with the top-dressing fertilizations. Maximum root yield (27.5 t ha-1) using 20-00-20 was obtained with the fertilization with 229.1 kg ha-1. However the joint application of nitrogen, potassium and calcium sources increased commercial root yield, especially of those classified as C-18, which is the major commercial class.

Avaliou-se o efeito de concentrações de P e épocas de coleta sobre o crescimento, nutrição mineral e teor de óleo essencial da menta (Mentha piperita L.). O delineamento experimental foi em blocos casualizados com parcelas subdivididas, em esquema fatorial 2x5, sendo 2 épocas de coleta (65 e 95 dias de cultivo (DC)) e 5 concentrações de P na solução nutritiva (6; 12; 18; 24 e 30 mg L-1), com 4 repetições. Foi determinada a massa seca de folhas (MSF), caules (MSC), raízes (MSR) e total (MST) e as relações folha:caule (F:C) e parte aérea:raiz (PA:R). Também, foi determinado o teor de óleo essencial em folhas frescas. Os níveis críticos foliares e acúmulo de nutrientes na parte aérea da menta foram estimados por meio da análise química das folhas e caules. As concentrações de P na solução influenciaram o crescimento e a produção de óleo essencial pela menta. Na coleta realizada aos 65 DC, apenas a MSF foi influenciada pelo P, não se observando efeito sobre os teores de óleo essencial. Aos 95 DC, o aumento da concentração de P promoveu aumentos significativos na massa seca das partes e do total da menta e, também, nos teores de óleo essencial, sendo o teor máximo (2,192 dag kg-1) obtido com 19,50 mg L-1 de P. As maiores concentrações de P avaliadas (24 e 30 mg L-1), promoveram aumentos na produção de matéria fresca e seca da parte aérea da menta, mas, reduziram os teores de óleo essencial, devido ao efeito de diluição, não significando, portanto, ganhos de produção de óleo por planta. Os teores foliares dos nutrientes associados ao teor máximo de óleo aos 95 DC foram em g kg-1: N=37,2; =3,9; K=21,2; Ca=9,3; Mg=3,8; S=3,0; e, em mg kg-1: B=35; Cu=8; Fe=323; Mn=145 e Zn=22. A exigência nutricional aos 95 DC relacionada ao teor máximo de óleo essencial seguiu a seqüência: N>K>Ca>P>Mg>S>Fe>Mn>B>Zn>Cu.

The effect of P concentrations and harvest period upon the growth, mineral nutrition and essential oil content of mint (Mentha piperita L.) was evaluated. The experimental design was of randomized blocks in a split-plot scheme, in a 2x5 factorial with four replications, being two harvest periods (65 and 95 days of cultivation (DC)) and five concentrations of P in the nutrient solution (6; 12; 18; 24 and 30 mg L-1) evaluated. The leaves dry weight (MSF), stems dry weight (MRS), roots dry weight (MSR), total dry weight (MST), leaf:stem ratio (F:C) and aerial part:root ratio (PA:R) were evaluated. The essential oil content in fresh leaves was determined. The critical levels and nutrient accumulation in leaves and stems were estimated through leaves and stems chemical analysis. The level of P increased the leaf dry matter production 65 DC. At 95 DC, the P concentration in the nutrient solution, did significantly increase plant growth and essential oil content, the maximum essential oil content being (2.192 dag kg-1) obtained with 19.48 mg L-1 of P. The higher P concentration in nutrient solution (24 and 30 mg L-1), increased the fresh and dry matter of the aerial portion of the plants, but, decreased the essential oil content. Due to the dilution effect, the increase of fresh and dry matter production did not result in an increase in the oil yield per plant. The critical levels of nutrients in the leaves corresponding to the maximum essential oil content at 95 DC in g kg-1 were: N=37,2; P=3,9; K=21,2; Ca=9,3; Mg=3,8; S=3,0 and, in mg kg-1: B=35; Cu=8; Fe=323; Mn=145 and Zn=22. The nutrients requested at 95 DC related to the maximum essential oil content was: N>K>Ca>P>Mg>S>Fe>Mn>B>Zn>Cu.

In this article are described new bioactive N-acylhydrazone (NAH) derivatives, structurally designed as optimization of aryl hydrazones precursors planned by molecular hybridization of two 5-lipoxigenase inhibitors, e.g. CBS-1108 and BW-755c. The analgesic, antiedematogenic and anti-platelet aggregating profile of several isosteric compounds was investigated by using classic pharmacological assays in vivo and ex-vivo, allowing to identify new potent peripheric analgesic lead, a new anti-inflammatory and an antithrombotic agent. During this study was discovered dozen of active NAH compounds clarifying the structure-activity relationship for this series of NAH derivatives, indicating the pharmacophore character of the N-acylhydrazone functionality.