Predicting the physicochemical profile of diastereoisomeric histidine-containing dipeptides by property space analysis.

OBJECTIVES: This study aimed at measuring the lipophilicity and ionization constants of diastereoisomeric dipeptides, interpreting them in terms of conformational behavior, and developing statistical models to predict them. METHODS: A series of 20 dipeptides of general structure NH(2) -L-X-(L or D)-His-OMe was designed and synthetized. Their experimental ionization constants (pK(1) , pK(2) and pK(3) ) and lipophilicity parameters (log P(N) and log D(7.4) ) were measured by potentiometry. Molecular modeling in three media (vacuum, water, and chloroform) was used to explore and sample their conformational space, and for each stored conformer to calculate their radius of gyration, virtual log P (preferably written as log P(MLP) , meaning obtained by the molecular lipophilicity potential (MLP) method) and polar surface area (PSA). Means and ranges were calculated for these properties, as was their sensitivity (i.e., the ratio between property range and number of rotatable bonds). RESULTS: Marked differences between diastereoisomers were seen in their experimental ionization constants and lipophilicity parameters. These differences are explained by molecular flexibility, configuration-dependent differences in intramolecular interactions, and accessibility of functional groups. Multiple linear equations correlated experimental lipophilicity parameters and ionization constants with PSA range and other calculated parameters. CONCLUSION: This study documents the differences in lipophilicity and ionization constants between diastereoisomeric dipeptides. Such configuration-dependent differences are shown to depend markedly on differences in conformational behavior and to be amenable to multiple linear regression. Chirality 24:566-576, 2012. © 2012 Wiley Periodicals, Inc.

Screening of the inhibitory effect of xenobiotics on alcohol metabolism using S9 rat liver fractions.

The purpose of this work was to develop and optimize a simple and suitable method to detect the potential inhibitory effect of drugs and medicines on alcohol dehydrogenase (ADH) activity in order to evaluate the possible interactions between medicines and alcohol metabolism. Commonly used medicines that are often involved in court litigations related with driving under the influence of alcohol were selected. Alprazolam, flunitrazepam and tramadol were tested as drugs with no known effect on ADH activity. Cimetidine, reported previously as having inhibitory effect on ADH, and 4-methylpyrazole (4-MP), a well known ADH inhibitor, were tested as positive controls. Apart from 4-MP, tramadol was identified as having the higher inhibitory effect with an IC50 of 44.7×10(-3)mM, followed by cimetidine (IC50 of 122.9×10(-3)mM). Alprazolam and flunitrazepam also reduced liver ADH activity but to a smaller extent (inhibition of 11.8±5.0% for alprazolam 1.0mM and 34.5±7.1% for flunitrazepam 0.04mM). Apart from cimetidine, this is the first report describing the inhibitory effect of these drugs on ethanol metabolism. The results also show the suitability of the method to screen for inhibitory effect of drugs on ethanol metabolism helping to identify drugs for which further study is justified.

Aspectos químicos e estruturais da qualidade fisiológica de sementes de soja

O objetivo deste trabalho foi avaliar os efeitos gênicos aditivos e não aditivos para as características da espessura das camadas da testa e da lignina presente nessas camadas, e verificar se os caracteres utilizados para avaliação da qualidade fisiológica das sementes de soja se correlacionam com o teor de lignina do tegumento. Foram usadas três cultivares (M-Soy 8400, CD201, CD206) de alta e três (CD215, CD202, Savana) de baixa qualidade fisiológica, para se obterem sementes híbridas F1 e F2, inclusive os recíprocos, pelo sistema de cruzamento dialelo parcial e as linhagens parentais. Os parâmetros primeira contagem de germinação, contagem final de germinação e velocidade de germinação, utilizados para avaliação da qualidade fisiológica, podem ser correlacionados com o teor de lignina. Foram observadas correlações positivas entre o teor de lignina e a percentagem de plântulas normais no teste de envelhecimento ao 5º e 11º dias após a semeadura, com valores maiores de correlação no 5º dia. A correlação entre velocidade de germinação e teor de lignina foi negativa. A análise do dialelo apontou significância do efeito recíproco, o que indica que a espessura das camadas da testa e a espessura de lignina nessas camadas podem ser explicadas pelo efeito materno.

Residual venous obstruction, alone and in combination with D-dimer, as a risk factor for recurrence after anticoagulation withdrawal following a first idiopathic deep vein thrombosis in the prolong study.

OBJECTIVE: This study aims to assess the predictive value of residual venous obstruction (RVO) for recurrent venous thrombo-embolism (VTE) in a study using D-dimer to predict outcome. DESIGN: This is a multicentre randomised open-label study. METHODS: Patients with a first episode of idiopathic VTE were enrolled on the day of anticoagulation discontinuation when RVO was determined by compression ultrasonography in those with proximal deep vein thrombosis (DVT) of the lower limbs. D-dimer was measured after 1 month. Patients with normal D-dimer did not resume anticoagulation while patients with abnormal D-dimer were randomised to resume anticoagulation or not. The primary outcome measure was recurrent VTE over an 18-month follow-up. RESULTS: A total of 490 DVT patients were analysed (after excluding 19 for different reasons and 118 for isolated pulmonary embolism (PE)). Recurrent DVT occurred in 19% (19/99) of patients with abnormal D-dimer who did not resume anticoagulation and 10% (31/310) in subjects with normal D-dimer (adjusted hazard ratio: 2.1; p = 0.02). Recurrences were similar in subjects either with (11%, 17/151) or without RVO (13%, 32/246). Recurrent DVT rates were also similar for normal D-dimer, with or without RVO, and for abnormal D-dimer, with or without RVO. CONCLUSIONS: Elevated D-dimer at 1 month after anticoagulation withdrawal is a risk factor for recurrence, while RVO at the time of anticoagulation withdrawal is not.

Reactions and enzymes in the metabolism of drugs and other xenobiotics.

In this article, we offer an overview of the compared quantitative importance of biotransformation reactions in the metabolism of drugs and other xenobiotics, based on a meta-analysis of current research interests. Also, we assess the relative significance the enzyme (super)families or categories catalysing these reactions. We put the facts unveiled by the analysis into a drug discovery context and draw some implications. The results confirm the primary role of cytochrome P450-catalysed oxidations and UDP-glucuronosyl-catalysed glucuronidations, but they also document the marked significance of several other reactions. Thus, there is a need for several drug discovery scientists to better grasp the variety of drug metabolism reactions and enzymes and their consequences.

Exploring the space of histidine containing dipeptides in search of novel efficient RCS sequestering agents.

The study reports a set of forty proteinogenic histidine-containing dipeptides as potential carbonyl quenchers. The peptides were chosen to cover as exhaustively as possible the accessible chemical space, and their quenching activities toward 4-hydroxy-2-nonenal (HNE) and pyridoxal were evaluated by HPLC analyses. The peptides were capped at the C-terminus as methyl esters or amides to favor their resistance to proteolysis and diastereoisomeric pairs were considered to reveal the influence of configuration on quenching. On average, the examined dipeptides are less active than the parent compound carnosine (βAla + His) thus emphasizing the unfavorable effect of the shortening of the βAla residue as confirmed by the control dipeptide Gly-His. Nevertheless, some peptides show promising activities toward HNE combined with a remarkable selectivity. The results emphasize the beneficial role of aromatic and positively charged residues, while negatively charged and H-bonding side chains show a detrimental effect on quenching. As a trend, ester derivatives are slightly more active than amides while heterochiral peptides are more active than their homochiral diastereoisomer. Overall, the results reveal that quenching activity strongly depends on conformational effects and vicinal residues (as evidenced by the reported QSAR analysis), offering insightful clues for the design of improved carbonyl quenchers and to rationalize the specific reactivity of histidine residues within proteins.

Qualitative structure-metabolism relationships in the hydrolysis of carbamates.

The aims of this review were 1) to compile a large number of reliable literature data on the metabolic hydrolysis of medicinal carbamates and 2) to extract from such data a qualitative relation between molecular structure and lability to metabolic hydrolysis. The compounds were classified according to the nature of their substituents (R³OCONR&supl;R²), and a metabolic lability score was calculated for each class. A trend emerged, such that the metabolic lability of carbamates decreased (i.e., their metabolic stability increased), in the following series: Aryl-OCO-NHAlkyl >> Alkyl-OCO-NHAlkyl ~ Alkyl-OCO-N(Alkyl)? ? Alkyl-OCO-N(endocyclic) ? Aryl-OCO-N(Alkyl)? ~ Aryl-OCO-N(endocyclic) ? Alkyl-OCO-NHAryl ~ Alkyl-OCO-NHAcyl?>> Alkyl-OCO-NH? > Cyclic carbamates. This trend should prove useful in the design of carbamates as drugs or prodrugs.

Caracterização morfológica da semente e da plântula de bacurizinho (Rheedia acuminata (Ruiz et Pav.) Plachon et Triana - Clusiaceae)

O bacurizinho é uma espécie frutífera do gênero Rheedia, nativa da Amazônia, e raramente cultivada, pois seus frutos, embora comestíveis, apresentam polpa escassa. Os estudos com o bacurizinho visam a sua utilização como porta-enxerto, redutor de porte para outras espécies do gênero Rheedia e Garcinia que apresentam altura elevada, como o bacuripari (Rheedia macrophylla Planchon et Triana- Clusiaceae) e o mangostão (Garcinia mangostana L.). Este trabalho teve como objetivo caracterizar, morfologicamente, a semente e a plântula dessa espécie. A semente é exalbuminosa, bitegumentada, com a testa de coloração marrom, apresentando vários feixes vasculares bem visíveis, distinguindo-se por sua coloração um pouco mais clara. O hilo é arredondado, de coloração escura, com pequena região mais clara ao centro, correspondendo no ponto de entrada de feixe vascular. A micrópila situa-se próxima ao hilo, sobre uma pequena protuberância triangular. O embrião é anômalo, hipocotilar e é representado unicamente por um longo eixo hipocótilo-radícula, sendo os cotilédones diminutos, aparecendo apenas na extremidade superior no lado oposto da micrópila. Durante a germinação, ocorre primeiramente a emergência de delgada raiz primária, no pólo oposto onde será originado o epicótilo. Essa raiz cresce aproximadamente 5-7cm e cessa, fenecendo posteriormente. Por ocasião da emergência do epicótilo, ocorre ao mesmo tempo a formação de uma raiz adventícia, na base do epicótilo, bem mais robusta que a anterior e que se constituirá no sistema radicular da planta. Precedendo a abertura do primeiro par de metáfilos, desenvolvem-se no epicótilo três pares de catafilos opostos e de coloração esverdeada. A germinação é hipógea e a plântula do tipo criptocotiledonar.

Development of a hepatocyte hollow fiber bioreactor for the study of contrast agents by magnetic resonance imaging

Thanks to the continuous progress made in recent years, medical imaging has become an important tool in the diagnosis of various pathologies. In particular, magnetic resonance imaging (MRI) permits to obtain images with a remarkably high resolution without the use of ionizing radiation and is consequently widely applied for a broad range of conditions in all parts of the body. Contrast agents are used in MRI to improve tissue discrimination. Different categories of contrast agents are clinically available, the most widely used being gadolinium chelates. One can distinguish between extracellular gadolinium chelates such as Gd-DTPA, and hepatobiliary gadolinium chelates such as Gd-BOPTA. The latter are able to enter hepatocytes from where they are partially excreted into the bile to an extent dependent on the contrast agent and animal species. Due to this property, hepatobiliary contrast agents are particularly interesting for the MRI of the liver. Actually, a change in signal intensity can result from a change in transport functions signaling the presence of impaired hepatocytes, e.g. in the case of focal (like cancer) or diffuse (like cirrhosis) liver diseases. Although the excretion mechanism into the bile is well known, the uptake mechanisms of hepatobiliary contrast agents into hepatocytes are still not completely understood and several hypotheses have been proposed. As a good knowledge of these transport mechanisms is required to allow an efficient diagnosis by MRI of the functional state of the liver, more fundamental research is needed and an efficient MRI compatible in vitro model would be an asset. So far, most data concerning these transport mechanisms have been obtained by MRI with in vivo models or by a method of detection other than MRI with cellular or sub-cellular models. Actually, no in vitro model is currently available for the study and quantification of contrast agents by MRI notably because high cellular densities are needed to allow detection, and no metallic devices can be used inside the magnet room, which is incompatible with most tissue or cell cultures that require controlled temperature and oxygenation. The aim of this thesis is thus to develop an MRI compatible in vitro cellular model to study the transport of hepatobiliary contrast agents, in particular Gd-BOPTA, into hepatocytes directly by MRI. A better understanding of this transport and especially of its modification in case of hepatic disorder could permit in a second step to extrapolate this knowledge to humans and to use the kinetics of hepatobiliary contrast agents as a tool for the diagnosis of hepatic diseases.

Development of a shelling method to recover whole kernels of the Cutia nut (Couepia edulis)

The kernel of the cutia nut (castanha-de-cutia, Couepia edulis (Prance) Prance) of the western Amazon, which is consumed by the local population, has traditionally been extracted from the nut with a machete, a dangerous procedure that only produces kernels cut in half. A shelling off machine prototype, which produces whole kernels without serious risks to its operator, is described and tested. The machine makes a circular cut in the central part of the fruit shell, perpendicular to its main axis. Three ways of conditioning the fruits before cutting were compared: (1) control; (2) oven drying immediately prior to cutting; (3) oven drying, followed by a 24-hour interval before cutting. The time needed to extract and separate the kernel from the endocarp and testa was measured. Treatment 3 produced the highest output: 63 kernels per hour, the highest percentage of whole kernels (90%), and the best kernel taste. Kernel extraction with treatment 3 required 50% less time than treatment 1, while treatment 2 needed 38% less time than treatment 1. The proportion of kernels attached to the testa was 93%, 47%, and 8% for treatments 1, 2, and 3, respectively, and was the main reason for extraction time differences.

Predicting drug metabolism: experiment and/or computation?

Drug metabolism can produce metabolites with physicochemical and pharmacological properties that differ substantially from those of the parent drug, and consequently has important implications for both drug safety and efficacy. To reduce the risk of costly clinical-stage attrition due to the metabolic characteristics of drug candidates, there is a need for efficient and reliable ways to predict drug metabolism in vitro, in silico and in vivo. In this Perspective, we provide an overview of the state of the art of experimental and computational approaches for investigating drug metabolism. We highlight the scope and limitations of these methods, and indicate strategies to harvest the synergies that result from combining measurement and prediction of drug metabolism.

Poliembrionia e atributos morfológicos de sementes de porta-enxertos de citros

A poliembrionia é um caráter fundamental no melhoramento genético e na multiplicação comercial de porta-enxertos de citros. Este trabalho avaliou as relações entre poliembrionia e diferentes atributos morfológicos de sementes de 12 genótipos de citros selecionados como porta-enxertos. Foram analisadas 50 sementes por genótipo, quanto a massa, altura, largura, número de embriões, taxa de poliembrionia, grau de dificuldade de extração da testa, coloração da semente, estimada por um matiz das duas notas de cores predominantes, e razão de cor, calculada pela divisão entre a menor e a maior nota do matiz. Os resultados foram submetidos à análise de variância, comparando-se as médias pelo teste de Scott-Knott e determinando-se os coeficientes de correlação de Pearson entre as variáveis (P < 0,05). Os porta-enxertos apresentaram atributos de sementes distintos, podendo ser agrupados em três classes de poliembrionia, destacando-se três seleções de limoeiro 'Cravo' com poliembrionia inferior a 50%, citrandarins como totalmente poliembriônicos e outros híbridos de Poncirus trifoliata com taxas baixas a intermediárias. O grau de dificuldade de extração da testa foi maior nas sementes menores. Na maioria dos genótipos avaliados, o número de embriões por semente correlacionou- se positivamente à massa, altura e largura de sementes e negativamente à razão de cor, sugerindo que a análise de coloração de sementes inteiras pode ser ferramenta alternativa para uma avaliação visual rápida da ocorrência de poliembrionia.

Types of stereoselectivity in drug metabolism: a heuristic approach.

Stereochemical factors are known to play a significant role in the metabolism of drugs and other xenobiotics. Following Prelog's lead, types of metabolic stereoselectivity can be categorized as (i) substrate stereoselectivity (the differential metabolism of two or more stereoisomeric substrates) and (ii) product stereoselectivity (the differential formation of two or more stereoisomeric metabolites from a single substrate). Combinations of the two categories exist as (iii) substrate-product stereoselectivities, meaning that product stereoselectivity itself is substrate stereoselective. Here, published examples of metabolic stereoselectivities are examined in the light of these concepts. In parallel, a graphical scheme is presented with a view to facilitate learning and help researchers to solve classification problems.