Chemoenzymatic Synthesis of Luliconazole Mediated by Lipases

A straightforward chemoenzymatic synthesis of luli- conazole has been developed. The key step involved the preparation of the enantiomerically pure beta-halohydrin (1S)-2-chloro- 1-(2,4-dichlorophenyl)-1-ethanol through kinetic resolution of the corresponding racemic acetate. This was achieved by a hydrolytic approach, mediated by the lipase from Thermomyces lanuginosus or Novozym 435\uae. The latter enzyme proved to be a robust biocatalyst for the kinetic resolution, and the halohydrin was obtained with high selectivity (ee > 99%, E > 200) after just 15 min, at 45 \ub0C. It could be reused five times with maintenance of high values of both conversion and enantioselectivity. Subsequently, the (S)-beta-halohydrin was sub- jected to a mesylation reaction; the mesylated derivative re- acted with 1-cyanomethylimidazole in the presence of CS2 to give luliconazole in 43 % yield with >99 % ee

Lipase mediated enzymatic kinetic resolution of phenylethyl halohydrins acetates: A case of study and rationalization

Racemic phenylethyl halohydrins acetates containing several groups attached to the aromatic ring were resolved via hydrolysis reaction in the presence of lipase B from Candida antarctica (Novozym\uae 435). In all cases, the kinetic resolution was highly selective (E > 200) leading to the corresponding (S)-\u3b2-halohydrin with ee > 99 %. However, the time required for an ideal 50 % conversion ranged from 15 min for 2,4-dichlorophenyl chlorohydrin acetate to 216 h for 2-chlorophenyl bromohydrin acetate. Six chlorohydrins and five bromohydrins were evaluated, the latter being less reactive. For the \u3b2-brominated substrates, steric hindrance on the aromatic ring played a crucial role, which was not observed for the \u3b2-chlorinated derivatives. To shed light on the different reaction rates, docking studies were carried out with all the substrates using MD simulations. The computational data obtained for the \u3b2-brominated substrates, based on the parameters analysed such as NAC (near attack conformation), distance between Ser-O and carbonyl-C and oxyanion site stabilization were in agreement with the experimental results. On the other hand, the data obtained for \u3b2-chlorinated substrates suggested that physical aspects such as high hydrophobicity or induced change in the conformation of the enzymatic active site are more relevant aspects when compared to steric hindrance effects

Chemoenzymatic synthesis of (S)-Pindolol using lipases

A straightforward chemoenzymatic synthesis of (S)-Pindolol has been developed. The key step involved the enzymatic kinetic resolution of rac-2-acetoxy-1-(1H-indol-4-yloxy)-3-chloropropane with lipase from Pseudomonas fluorescens via hydrolytic process to obtain enantiomerically enriched halohydrin (2S)-1-(1H-indol-4-yloxy)-3-chloro-2-propanol (96% ee) and (2R)-2-acetoxy-1-(1H-indol-4-yloxy)-3-chloropropane (97% ee). The latter was subjected to a hydrolysis reaction catalyzed by Candida rugosa leading to (2R)-1-(1H-indol-4-yloxy)-3-chloro-2-propanol (97% ee), followed by a reaction with isopropylamine, producing (S)-Pindolol (97% ee) in quantitative yield

Lipase mediated enzymatic kinetic resolution of phenylethyl halohydrins acetates: A case of study and rationalization

11siRacemic phenylethyl halohydrins acetates containing several groups attached to the aromatic ring were resolved via hydrolysis reaction in the presence of lipase B from Candida antarctica (Novozym® 435). In all cases, the kinetic resolution was highly selective (E>200) leading to the corresponding (S)-β-halohydrin with ee>99 %. However, the time required for an ideal 50 % conversion ranged from 15 min for 2,4-dichlorophenyl chlorohydrin acetate to 216 h for 2-chlorophenyl bromohydrin acetate. Six chlorohydrins and five bromohydrins were evaluated, the latter being less reactive. For the β-brominated substrates, steric hindrance on the aromatic ring played a crucial role, which was not observed for the β-chlorinated derivatives. To shed light on the different reaction rates, docking studies were carried out with all the substrates using MD simulations. The computational data obtained for the β-brominated substrates, based on the parameters analysed such as NAC (near attack conformation), distance between Ser-O and carbonyl-C and oxyanion site stabilization were in agreement with the experimental results. On the other hand, the data obtained for β-chlorinated substrates suggested that physical aspects such as high hydrophobicity or induced change in the conformation of the enzymatic active site are more relevant aspects when compared to steric hindrance effects.partially_openopenThiago de Sousa Fonseca, Kimberly Benedetti Vega, Marcos Reinaldo da Silva, Maria da Conceição Ferreira de Oliveira, Telma Leda Gomes de Lemosa, Martina Letizia Contenteb, Francesco Molinari, Marco Cespugli, Sara Fortuna, Lucia Gardossi, Marcos Carlos de Mattos,de Sousa Fonseca, Thiago; Benedetti Vega, Kimberly; Reinaldo da Silva, Marcos; da Conceição Ferreira de Oliveira, Maria; Leda Gomes de Lemosa, Telma; Letizia Contenteb, Martina; Molinari, Francesco; Cespugli, Marco; Fortuna, Sara; Gardossi, Lucia; Carlos de Mattos, Marco

Ferulic acid lowers body weight and visceral fat accumulation via modulation of enzymatic, hormonal and inflammatory changes in a mouse model of high-fat diet-induced obesity

Previous studies have reported on the glucose and lipid-lowering effects of ferulic acid (FA) but its anti-obesity potential has not yet been firmly established. This study investigated the possible anti-obesitogenic effects of FA in mice fed a high-fat diet (HFD) for 15 weeks. To assess the antiobesity potential of FA, 32 male Swiss mice, weighing 20–25 g (n=6–8 per group) were fed a normal diet (ND) or HFD, treated orally or not with either FA (10 mg/kg) or sibutramine (10 mg/kg) for 15 weeks and at the end of this period, the body weights of animals, visceral fat accumulation, plasma levels of glucose and insulin hormone, amylase and lipase activities, the satiety hormones ghrelin and leptin, and tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCH-1) were analyzed. Results revealed that FA could effectively suppress the HFD-associated increase in visceral fat accumulation, adipocyte size and body weight gain, similar to sibutramine, the positive control. FA also significantly (P<0.05) decreased the HFD-induced elevations in serum lipid profiles, amylase and lipase activities, and the levels of blood glucose and insulin hormone. The markedly elevated leptin and decreased ghrelin levels seen in HFD-fed control mice were significantly (P<0.05) reversed by FA treatment, almost reaching the values seen in ND-fed mice. Furthermore, FA demonstrated significant (P<0.05) inhibition of serum levels of inflammatory mediators TNF-α, and MCH-1. These results suggest that FA could be beneficial in lowering the risk of HFD-induced obesity via modulation of enzymatic, hormonal and inflammatory responses

Teor de óleo essencial de alecrim-pimenta em função do horário de colheita Essential oil content of pepper-rosmarin as a function of harvest time

O alecrim-pimenta é uma importante planta medicinal, nativa do nordeste brasileiro, que, devido ao seu óleo essencial, apresenta grande potencial antifúngico e antibacteriano. Um dos fatores que afetam o teor de metabólitos secundários das plantas é o horário de colheita, assim, o conhecimento desse fator subsidia as decisões agronômicas da espécie, favorecendo o aumento do teor de óleo essencial no momento da colheita. Dessa forma, o objetivo do presente trabalho foi avaliar o efeito do horário de colheita na produção de óleo essencial de alecrim-pimenta (Lippia sidoides Cham.). O experimento foi desenvolvido no Instituto de Ciências Agrárias da Universidade Federal de Minas Gerais (ICA/UFMG), Montes Claros, Minas Gerais, Brasil. Utilizou-se o delineamento experimental em blocos casualizados com cinco tratamentos, definidos pelos horários de coleta das folhas (6:00, 9:00, 12:00, 15:00 e 18:00) e seis repetições, sendo esse o número de coletas em cada tratamento. As folhas foram obtidas de plantas matrizes do Horto Medicinal do ICA/UFMG. O óleo essencial foi obtido pelo método de hidrodestilação usando o aparelho de Clevenger. A análise de regressão apresentou um efeito cúbico para o teor de óleo essencial, sendo que o maior valor, 5,78% em relação à matéria seca, foi obtido às 9h59min. Portanto, recomenda-se a colheita das folhas de alecrim-pimenta próximo das 10h.<br>The pepper-rosmarin is an important medicinal plant, native of Northeast of Brazil, that, because of its essential oil, presents great antifungal and antibacterial potential. One of the factors that affect the secondary metabolites content of plants is the harvest time, thus, the knowledge of this factor subsidizes the decisions agronomic species, favoring the increase of essential oil content at the harvest moment. Thus, the aim of this research was to evaluate the effect of harvest time on the essential oil production of pepper-rosmarin (Lippia sidoides Cham.). The experiment was carried out in the Instituto of Ciências Agrárias of Universidade Federal of Minas Gerais (ICA/UFMG), Montes Claros, Minas Gerais, Brazil. It was installed in a randomized block design, with five treatments consisted by harvest times (6:00, 9:00, 12:00, 15:00 and 18:00) with six replications (number of collections in each treatment). The leaves were obtained from stock plants of medicinal vegetable garden of ICA/ UFMG. The essential oil was obtained by hydrodistillation in a Clevenger apparatus. The regression analysis presented a cubic effect to the essential oil content, being that the highest value, 5,78% in relation to dry matter, was obtained at 9h59min. Therefore, it was recommended the harvest time of pepper-rosmarin leaves next to 10h