5-Fluorouracil(5-FU)is an anticancer drug extensively used for different cancers.Intracellular metabolic activation leads to several nucleoside and nucleotide metabolites essential to exert its cytotoxic activity on m...5-Fluorouracil(5-FU)is an anticancer drug extensively used for different cancers.Intracellular metabolic activation leads to several nucleoside and nucleotide metabolites essential to exert its cytotoxic activity on multiple cellular targets such as enzymes,DNA and RNA.In this paper,we describe the development of a method based on liquid chromatography coupled with high resolution mass spectrometry suitable for the simultaneous determination of the ten anabolic metabolites(nucleoside,nucleotide and sugar nucleotide)of 5-FU.The chromatographic separationwas optimized on a porous graphitic carbon column allowing the analysis of the metabolites of 5-FU as well as endogenous nucleotides.The detection was performed on an Orbitrap■tandem mass spectrometer.Linearity of the method was verified in intracellular content and in RNA extracts.The limit of detection was equal to 12 pg injected on column for nucleoside metabolites of 5-FU and 150 pg injected on column for mono-and tri-phosphate nucleotide metabolites.Matrix effect was evaluated in cellular contents,DNA and RNA extracts for nucleoside and nucleotides metabolites.The method was successfully applied to i)measure the proportion of each anabolic metabolite of 5-FU in cellular contents,ii)follow the consequence of inhibition of enzymes on the endogenous nucleotide pools,iii)study the incorporation of metabolites of 5-FU into RNA and DNA,and iv)to determine the incorporation rate of 5-FUrd into 18 S and 28 S sub-units of rRNA.展开更多
A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination...A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination provided enzalutamide in 41.0%total yield.This route avoids the using of toxic chemical,unstable intermediate and high-risk reaction.It is a potential efficient and economical procedure for industrialization.展开更多
The outbreak of oxidative stress, inflammatory diseases and mycoses, constitute an important public health problem. This could be due to the increase of risk factors, side effects and expensive therapeutic molecules a...The outbreak of oxidative stress, inflammatory diseases and mycoses, constitute an important public health problem. This could be due to the increase of risk factors, side effects and expensive therapeutic molecules available. In the aim to find the potential spring of new therapeutic molecules with efficient and wide spectrum of action, the antiradical, anti-inflammatory and antifungal activities of two essential oils were evaluated. The essential oils were extracted by hydrodistillation. The antiradical activity was evaluated to use DPPH （2,2-diphenyl-l-picrylhydrazyl） scavenging method and the anti-inflammatory activity was determined using the enzymatic method. The disc diffusion and dilution methods were used to evaluate growth inhibition of three yeasts, three moulds and three dermatophytes. The yields of extraction of Apium graveolens and Thymus vulgaris essential oils were 0.14% and 0.32% respectively. These essential oils showed antiradical properties with respective SC50 of 0.41 and 0.06 g/L for Apium graveolens and Thymus vulgaris. Only Thymus vulgaris presented an anti-inflammatory activity with an IC50 of 0.19 g/L. Cryptococcus neoformans was the most susceptible fungal strain while C. albicans was the most resistant one. The results were compared with the standard antifungal. These results show that these essential oils could be exploited as potential spring of molecules endowed with antiradical, anti-inflammatory and antifungal activities.展开更多
Autism spectrum disorders (ASDs) are epidemically explosive clinical entities, but their pathogenesis is still unclear and a definitive cure does not yet exist. Rett syndrome (RTT) is a rare genetically determined cau...Autism spectrum disorders (ASDs) are epidemically explosive clinical entities, but their pathogenesis is still unclear and a definitive cure does not yet exist. Rett syndrome (RTT) is a rare genetically determined cause of autism linked to mutations in the X-linked MeCP2 gene or, more rarely, in CDKL5 or FOXG1. A wide phenotypical heterogeneity is a known feature of the disease. Although several studies have focused on the molecular genetics and possible protein changes at different levels, to date very little attention has been paid to fatty acids in this disease, which could be considered as a natural paradigm for the ASDs. To this regard, a quite enigmatic feature of the disease is the evidence in the affected patients of an extensive peroxidation of polyunsaturated fatty acids (arachidonic acid, AA, docosaexahenoic acid, DHA, adrenic acid, AdA and, to a lesser extent, eicosapentaenoic acid, EPA), in contrast with amelioration of the redox changes and phenotypical severity following the supplementation of some of those same fatty acids (DHA + EPA). Therefore, fatty acids may represent a kind of Janus Bifrons in the particular context of RTT. Here, we propose a rational explanation for this apparent “fatty acid paradox” in RTT. A better understanding of this paradox could also be of help to get a better insight into the complex mechanism of action for polyunsaturated fatty acids in health and disease.展开更多
Gram‐negative bacteria are particularly prone to developing antimicrobial resistance(AMR),as evidenced by the WHO's ESKAPEE list of high‐priority pathogens.One strategy that has increased is the use of antibioti...Gram‐negative bacteria are particularly prone to developing antimicrobial resistance(AMR),as evidenced by the WHO's ESKAPEE list of high‐priority pathogens.One strategy that has increased is the use of antibiotic enhancers,which can reempower abandoned or poorly active antibiotics against the resistant strain of interest.In this study,the polyamino‐isoprenyl antibiotic enhancer,NV716,was tested in combination with two families of multi‐target Ser/Cys‐based enzyme inhibitors,the oxadiazolone derivatives(OX)and the Cyclipostins and Cyclophostin analogs(CyC),which are inactive against Gram‐negative ESKAPEE bacteria,to potentiate their antibacterial activity and thus make them active against these bacteria.We demonstrated that NV716 potentiates some OX and CyC compounds by permeabilizing the outer membrane and thus by increasing the inhibitor accumulation,as shown by fluorescence microscopy.By using the click‐chemistry activity‐based protein profiling(ABPP)approach coupled with proteomic analysis,we also confirmed the multi‐target nature of the best OX and CyC inhibitors by identifying their target proteins on a bacterial culture of Enterobacter cloacae.Remarkably,a large set of these identified proteins had already been captured in previous ABPP experiments conducted on Mycobacterium tuberculosis and/or Mycobacterium abscessus culture.Furthermore,we showed that five of the identified target proteins were present in a total lysate of Pseudomonas aeruginosa.Importantly,these latter enzymes are highly conserved among Gram‐negative bacteria,with two of them annotated as essential for bacterial survival.These results provide proof of concept that both OX and CyC,if successfully potentiated,could be used against ESKAPEE Gram‐negative bacteria.展开更多
基金supported by Institut National Du Cancer(grant number 2018-131).
文摘5-Fluorouracil(5-FU)is an anticancer drug extensively used for different cancers.Intracellular metabolic activation leads to several nucleoside and nucleotide metabolites essential to exert its cytotoxic activity on multiple cellular targets such as enzymes,DNA and RNA.In this paper,we describe the development of a method based on liquid chromatography coupled with high resolution mass spectrometry suitable for the simultaneous determination of the ten anabolic metabolites(nucleoside,nucleotide and sugar nucleotide)of 5-FU.The chromatographic separationwas optimized on a porous graphitic carbon column allowing the analysis of the metabolites of 5-FU as well as endogenous nucleotides.The detection was performed on an Orbitrap■tandem mass spectrometer.Linearity of the method was verified in intracellular content and in RNA extracts.The limit of detection was equal to 12 pg injected on column for nucleoside metabolites of 5-FU and 150 pg injected on column for mono-and tri-phosphate nucleotide metabolites.Matrix effect was evaluated in cellular contents,DNA and RNA extracts for nucleoside and nucleotides metabolites.The method was successfully applied to i)measure the proportion of each anabolic metabolite of 5-FU in cellular contents,ii)follow the consequence of inhibition of enzymes on the endogenous nucleotide pools,iii)study the incorporation of metabolites of 5-FU into RNA and DNA,and iv)to determine the incorporation rate of 5-FUrd into 18 S and 28 S sub-units of rRNA.
基金supported by Scientific Research Cadre Project(AMSCP)2021National Key Research and Development Program of China(No.2020YFA0509200 to C.Sheng)。
文摘A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination provided enzalutamide in 41.0%total yield.This route avoids the using of toxic chemical,unstable intermediate and high-risk reaction.It is a potential efficient and economical procedure for industrialization.
文摘The outbreak of oxidative stress, inflammatory diseases and mycoses, constitute an important public health problem. This could be due to the increase of risk factors, side effects and expensive therapeutic molecules available. In the aim to find the potential spring of new therapeutic molecules with efficient and wide spectrum of action, the antiradical, anti-inflammatory and antifungal activities of two essential oils were evaluated. The essential oils were extracted by hydrodistillation. The antiradical activity was evaluated to use DPPH （2,2-diphenyl-l-picrylhydrazyl） scavenging method and the anti-inflammatory activity was determined using the enzymatic method. The disc diffusion and dilution methods were used to evaluate growth inhibition of three yeasts, three moulds and three dermatophytes. The yields of extraction of Apium graveolens and Thymus vulgaris essential oils were 0.14% and 0.32% respectively. These essential oils showed antiradical properties with respective SC50 of 0.41 and 0.06 g/L for Apium graveolens and Thymus vulgaris. Only Thymus vulgaris presented an anti-inflammatory activity with an IC50 of 0.19 g/L. Cryptococcus neoformans was the most susceptible fungal strain while C. albicans was the most resistant one. The results were compared with the standard antifungal. These results show that these essential oils could be exploited as potential spring of molecules endowed with antiradical, anti-inflammatory and antifungal activities.
文摘Autism spectrum disorders (ASDs) are epidemically explosive clinical entities, but their pathogenesis is still unclear and a definitive cure does not yet exist. Rett syndrome (RTT) is a rare genetically determined cause of autism linked to mutations in the X-linked MeCP2 gene or, more rarely, in CDKL5 or FOXG1. A wide phenotypical heterogeneity is a known feature of the disease. Although several studies have focused on the molecular genetics and possible protein changes at different levels, to date very little attention has been paid to fatty acids in this disease, which could be considered as a natural paradigm for the ASDs. To this regard, a quite enigmatic feature of the disease is the evidence in the affected patients of an extensive peroxidation of polyunsaturated fatty acids (arachidonic acid, AA, docosaexahenoic acid, DHA, adrenic acid, AdA and, to a lesser extent, eicosapentaenoic acid, EPA), in contrast with amelioration of the redox changes and phenotypical severity following the supplementation of some of those same fatty acids (DHA + EPA). Therefore, fatty acids may represent a kind of Janus Bifrons in the particular context of RTT. Here, we propose a rational explanation for this apparent “fatty acid paradox” in RTT. A better understanding of this paradox could also be of help to get a better insight into the complex mechanism of action for polyunsaturated fatty acids in health and disease.
基金supported by the CNRS,INSERM,and Aix Marseille University.Proteomics analyses were carried out using the mass spectrometry facility of Marseille Proteomics(marseille‐proteomique.univ‐amu.fr)supported by IBISAthe Cancéropôle PACA,the Provence‐Alpes‐Côte d'Azur Region,the Institut Paoli‐Calmettes,and Fonds Européen de Développement Regional(FEDER)+1 种基金Emma Forest PhD fellowship is supported by the Ministère de l'Enseignement Supérieur et de la RechercheThe authors would like to thank Pierre Santucci(LISM UMR7255 CNRS)for his help with Biorender.
文摘Gram‐negative bacteria are particularly prone to developing antimicrobial resistance(AMR),as evidenced by the WHO's ESKAPEE list of high‐priority pathogens.One strategy that has increased is the use of antibiotic enhancers,which can reempower abandoned or poorly active antibiotics against the resistant strain of interest.In this study,the polyamino‐isoprenyl antibiotic enhancer,NV716,was tested in combination with two families of multi‐target Ser/Cys‐based enzyme inhibitors,the oxadiazolone derivatives(OX)and the Cyclipostins and Cyclophostin analogs(CyC),which are inactive against Gram‐negative ESKAPEE bacteria,to potentiate their antibacterial activity and thus make them active against these bacteria.We demonstrated that NV716 potentiates some OX and CyC compounds by permeabilizing the outer membrane and thus by increasing the inhibitor accumulation,as shown by fluorescence microscopy.By using the click‐chemistry activity‐based protein profiling(ABPP)approach coupled with proteomic analysis,we also confirmed the multi‐target nature of the best OX and CyC inhibitors by identifying their target proteins on a bacterial culture of Enterobacter cloacae.Remarkably,a large set of these identified proteins had already been captured in previous ABPP experiments conducted on Mycobacterium tuberculosis and/or Mycobacterium abscessus culture.Furthermore,we showed that five of the identified target proteins were present in a total lysate of Pseudomonas aeruginosa.Importantly,these latter enzymes are highly conserved among Gram‐negative bacteria,with two of them annotated as essential for bacterial survival.These results provide proof of concept that both OX and CyC,if successfully potentiated,could be used against ESKAPEE Gram‐negative bacteria.
