RESUMEN
A molecularly imprinted polymer of Tenofovir (1), an FDA-approved acyclic nucleoside phosphonate with antiviral activity, was synthesized using a non-covalent approach. A pre-polymerization complex was formed between (1) and DMAEMA and in-house synthetic N1-[(2-methacryloyloxy)ethyl] thymine, with EGDMA as a cross-linker in an MeCN/H2O (9:1, 1:1) mixture as a porogen, giving an imprinting factor (IF) of 5.5 at 2.10-5 mol/L. Binding parameters were determined by the Freundlich-Langmuir model, Qmax and Ka, and well as the particle morphology for MIP and NIP. Finally, the release profiles, for MIP and NIP, were obtained at 25 °C and 37 °C, which is body temperature, in a phosphate buffer saline, pH 7.4, mimicking the blood pH value, to determine the potential sustained release of our polymeric materials.
RESUMEN
New acyclic pyrimidine nucleoside phosphonate prodrugs with a 4-(2,4-diaminopyrimidin-6-yl)oxy-but-2-enyl]phosphonic acid skeleton (O-DAPy nucleobase) were prepared through a convergent synthesis by olefin cross-metathesis as the key step. Several acyclic nucleoside 4-(2,4-diaminopyrimidin-6-yl)oxy-but-2-enyl]phosphonic acid prodrug exhibited in vitro antiviral activity in submicromolar or nanomolar range against varicella zoster virus (VZV), human cytomegalovirus (HCMV), human herpes virus type 1 (HSV-1) and type 2 (HSV-2), and vaccinia virus (VV), with good selective index (SI). Among them, the analogue 9c (LAVR-289) proved markedly inhibitory against VZV wild-type (TK+) (EC50 0.0035 µM, SI 740) and for thymidine kinase VZV deficient strains (EC50 0.018 µM, SI 145), with a low morphological toxicity in cell culture at 100 µM and acceptable cytostatic activity resulting in excellent selectivity. Compound 9c exhibited antiviral activity against HCMV (EC50 0.021 µM) and VV (EC50 0.050 µM), as well as against HSV-1 (TK-) (EC50 0.0085 µM). Finally, LAVR-289 (9c) deserves further (pre)clinical investigations as a potent candidate broad-spectrum anti-herpesvirus drug.
Asunto(s)
Antivirales , Virus ADN , Pruebas de Sensibilidad Microbiana , Profármacos , Antivirales/farmacología , Antivirales/síntesis química , Antivirales/química , Profármacos/farmacología , Profármacos/síntesis química , Profármacos/química , Humanos , Virus ADN/efectos de los fármacos , Relación Estructura-Actividad , Herpesvirus Humano 1/efectos de los fármacos , Estructura Molecular , Herpesvirus Humano 3/efectos de los fármacos , Organofosfonatos/farmacología , Organofosfonatos/química , Organofosfonatos/síntesis química , Citomegalovirus/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Virus Vaccinia/efectos de los fármacos , Herpesvirus Humano 2/efectos de los fármacosRESUMEN
The emergence of Multidrug Resistance (MDR) strains of bacteria has accelerated the search for new antibacterials. The specific bacterial peptidoglycan biosynthetic pathway represents opportunities for the development of novel antibacterial agents. Among the enzymes involved, Mur ligases, described herein, and especially the amide ligases MurC-F are key targets for the discovery of multi-inhibitors, as they share common active sites and structural features.
Asunto(s)
Antibacterianos , Ligasas , Ligasas/metabolismo , Antibacterianos/farmacología , Antibacterianos/química , Bacterias/metabolismo , Dominio Catalítico , Farmacorresistencia Microbiana , PeptidoglicanoRESUMEN
In 2002, a new class of thymidylate synthase (TS) involved in the de novo synthesis of dTMP named Flavin-Dependent Thymidylate Synthase (FDTS) encoded by the thyX gene was discovered; FDTS is present only in 30% of prokaryote pathogens and not in human pathogens, which makes it an attractive target for the development of new antibacterial agents, especially against multi-resistant pathogens. We report herein the synthesis and structure-activity relationship of a novel series of hitherto unknown pyrido[1,2-e]purine-2,4(1H,3H)-dione analogues. Several synthetics efforts were done to optimize regioselective N1-alkylation through organopalladium cross-coupling. Modelling of potential hits were performed to generate a model of interaction into the active pocket of FDTS to understand and guide further synthetic modification. All those compounds were evaluated on an in-house in vitro NADPH oxidase assays screening as well as against Mycobacterium tuberculosis ThyX. The highest inhibition was obtained for compound 23a with 84.3% at 200 µM without significant cytotoxicity (CC50 > 100 µM) on PBM cells.
Asunto(s)
Mycobacterium tuberculosis , Antibacterianos/farmacología , Dinitrocresoles , Flavinas/metabolismo , Flavinas/farmacología , Humanos , Mycobacterium tuberculosis/genética , NADPH Oxidasas , Purinas/farmacología , Relación Estructura-Actividad , Timidina Monofosfato , Timidilato Sintasa/metabolismoRESUMEN
The synthesis of (1R)-2-amino-2-deoxy-ß-l-gulopyranosyl benzene and the α and ß forms of 2-amino-2-deoxy-l-idopyranosyl benzene derivatives was accomplished through stereospecific addition of tributylstannyllithium to readily available (SR)- or (SS)-N-tert-butanesulfinyl-arabinofuranosylamine building blocks, followed by stereoretentive Pd-catalyzed Migita-Kosugi-Stille cross-coupling, stereoselective reduction, and an activation-cyclization strategy. Application of this methodology paves the way to new three-dimensional chemical space and preparation of unknown (non-natural) and complex 2-amino-2-deoxy sugars of biological interest.
Asunto(s)
Desoxiazúcares , Paladio , Benceno , Ciclización , EstereoisomerismoRESUMEN
With several US Food and Drug Administration (FDA)-approved drugs and high barriers to resistance, nucleoside and nucleotide analogs remain the cornerstone of antiviral therapies for not only herpesviruses, but also HIV and hepatitis viruses (B and C); however, with the exception of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which vaccines have been developed at unprecedented speed, there are no vaccines or small antivirals yet available for (re)emerging viruses, which are primarily RNA viruses. Thus, herein, we present an overview of ribonucleoside analogs recently developed and acting as inhibitors of the viral RNA-dependent RNA polymerase (RdRp). They are new lead structures that will be exploited for the discovery of new antiviral nucleosides.
Asunto(s)
Antivirales , Nucleósidos , Antivirales/química , Antivirales/farmacología , Humanos , Nucleósidos/análogos & derivados , Nucleósidos/farmacología , ARN Polimerasa Dependiente del ARN/antagonistas & inhibidores , Estados UnidosRESUMEN
Nucleoside analogues are at the forefront of antiviral therapy for last decades. To circumvent some of their limitations, based on their metabolism, and in order to improve their anti-viral potency and selectivity, several families of nucleoside analogues have been described through structural modifications at the sugar and heterocycles. The replacement of the oxygen of the nucleoside by a methylene has led to the family of carbocyclic (or cyclopentane) nucleoside analogues. Various potent anti-HIV and anti-HBV drugs belong to this family. Main syntheses of carbocyclic analogues of nucleosides used Diels-Alder reactions (in racemic or asymmetric series), but also started from carbohydrates (ribose, glucose), as a source of optically active compounds, which then had to be transformed into carbacycles under various conditions. The growing interest in carbocyclic nucleosides has led several groups, including ours, to develop new analogues and to explore novel routes. This article will review some of the recent chemistry developed on selected five-membered ring carbocyclic nucleosides.
Asunto(s)
Antivirales , Nucleósidos , Antivirales/química , Antivirales/farmacología , Reacción de Cicloadición , Nucleósidos/química , Nucleósidos/farmacologíaRESUMEN
This study describes the synthesis and evaluation of different imprinted hydrogels using ribavirin as template molecule. Ribavirin serves as a model molecule because it possesses a broad-spectrum antiviral effect against RNA viruses, which are expected as emerging viruses. The choice of monomers enables to stabilize the pre-polymerization complex and to synthesize biocompatible polymers. Predictive studies as well as experimental works conclude similar results on best ribavirin:monomers ratios. Thus, materials exhibit high selective cavities toward ribavirin. These affinities allow to show release profiles drastically different from the non-imprinted ones at two temperatures. The imprinted materials show a sustained profile able to release antiviral for more than 24 h. The hydrogels obtained are biocompatible with model cells retained, human lung epithelial BEAS-2B cells. Cell viability is excellent and pro-inflammatory response is insignificant when imprinted polymers are incubated with cells. Finally, viral tests carried out on Influenza A infected lung cells show that imprinted delivery systems delivering 1 to 3 µg of antiviral have the same efficiency as a medium containing 30 µg mL-1 of active agent. As a very interesting result, the molecularly imprinted polymers as drug delivery systems allow to increase the local concentration of antiviral, to improve their delivery when its bioavailability is low.
Asunto(s)
Virus de la Influenza A , Impresión Molecular , Antivirales/farmacología , Sistemas de Liberación de Medicamentos , Humanos , Hidrogeles/farmacología , Impresión Molecular/métodos , Nucleósidos , Ribavirina/farmacologíaRESUMEN
Flavin-Dependent Thymidylate Synthase (FDTS) encoded by ThyX gene was discovered as a new class of thymidylate synthase involved in the de novo synthesis of dTMP named only in 30 % of human pathogenic bacteria. This target was pursed for the development of new antibacterial agents against multiresistant pathogens. We have developed a new class of ANPs based on the mimic of two natural's cofactors (dUMP and FAD) as inhibitors against Mycobacterium tuberculosis ThyX. Several synthetic efforts were performed to optimize regioselective N1-alkylation, cross-coupling metathesis and Sonogashira cross-coupling. Compound 19c showed a poor 31.8% inhibitory effect on ThyX at 200 µM.
Asunto(s)
Antibacterianos/farmacología , Mycobacterium tuberculosis/efectos de los fármacos , Nucleósidos/farmacología , Timidilato Sintasa/antagonistas & inhibidores , Antibacterianos/síntesis química , Antibacterianos/química , Relación Dosis-Respuesta a Droga , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Mycobacterium tuberculosis/enzimología , Nucleósidos/síntesis química , Nucleósidos/química , Relación Estructura-Actividad , Timidilato Sintasa/metabolismoRESUMEN
A series of hitherto unknown (1,4-disubstituted-1,2,3-triazol)-(E)-2-methyl-but-2-enyl nucleosides phosphonate prodrugs bearing 4-substituted-1,2,3-triazoles were prepared in a straight approach through an olefin acyclic cross metathesis as the key synthetic step. All novel compounds were evaluated for their antiviral activities against HBV, HIV and SARS-CoV-2. Among these molecules, only compound 15j, a hexadecyloxypropyl (HDP)/(isopropyloxycarbonyl-oxymethyl)-ester (POC) prodrug, showed activity against HBV in Huh7 cell cultures with 62% inhibition at 10 µM, without significant cytotoxicity (IC50 = 66.4 µM in HepG2 cells, IC50 = 43.1 µM in HepG2 cells) at 10 µM.
Asunto(s)
Antivirales/síntesis química , Antivirales/farmacología , Compuestos Azo/química , Nucleósidos/química , Organofosfonatos/química , Profármacos/síntesis química , Profármacos/farmacología , Alquenos/química , Animales , Línea Celular Tumoral , Chlorocebus aethiops , VIH-1/efectos de los fármacos , Virus de la Hepatitis B/efectos de los fármacos , Humanos , Espectroscopía de Resonancia Magnética , Metilación , SARS-CoV-2/efectos de los fármacos , Relación Estructura-Actividad , Triazoles/química , Células VeroRESUMEN
Novel 2-substituted-6-[(4-substituted-1-piperidyl)methyl]-1H-benzimidazoles were designed and synthesized as Ebola virus inhibitors. The proposed structures of the new prepared benzimidazole-piperidine hybrids were confirmed based on their spectral data and CHN analyses. The target compounds were screened in vitro for their anti-Ebola activity. Among tested molecules, compounds 26a (EC50=0.93 µM, SI = 10) and 25a (EC50=0.64 µM, SI = 20) were as potent as and more selective than Toremifene reference drug (EC50 = 0.38 µM, SI = 7) against cell line. Data suggests that the mechanism by which 25a and 26a block EBOV infection is through the inhibition of viral entry at the level of NPC1. Furthermore, a docking study revealed that several of the NPC1 amino acids that participate in binding to GP are involved in the binding of the most active compounds 25a and 26a. Finally, in silico ADME prediction indicates that 26a is an idealy drug-like candidate. Our results could enable the development of small molecule drug capable of inhibiting Ebola virus, especially at the viral entry step.
Asunto(s)
Antivirales/farmacología , Bencimidazoles/farmacología , Diseño de Fármacos , Fiebre Hemorrágica Ebola/tratamiento farmacológico , Antivirales/síntesis química , Antivirales/química , Bencimidazoles/síntesis química , Bencimidazoles/química , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células HeLa , Humanos , Estructura Molecular , Relación Estructura-ActividadRESUMEN
The synthesis of hitherto unknown 5'-deoxy-5'-(4-substituted-1,2,3-triazol-1-yl)-uridine and its evaluation, through an one-pot screening assay, against MurA-F enzymes involved in Mycobacterium tuberculosis (Mtb), are described. Starting from UDP-N-acetylmuramic acid (UDP-MurNAc), the natural substrate involved in the peptidoglycan biosynthesis, our strategy was to substitute the diphosphate group of UDP-MurNAc by a 1,2,3-triazolo spacer under copper-catalyzed azide-alkyne cycloaddition conditions. The structure-activity relationship was discussed and among the 23 novel compounds developed, N-acetylglucosamine analogues 11c and 11e emerged as the best inhibitors against the Mtb MurA-F enzymes reconstruction pathway with an inhibitory effect of 56% and 50%, respectively, at 100 µM. Both compounds are selective inhibitors of Mtb MurE, the molecular docking and molecular dynamic simulation suggesting that 11c and 11e are occupying the active site of Mtb MurE ligase.
Asunto(s)
Diseño de Fármacos , Mycobacterium tuberculosis/enzimología , Péptido Sintasas/antagonistas & inhibidores , Triazoles/química , Uridina/síntesis química , Uridina/farmacología , Dominio Catalítico , Técnicas de Química Sintética , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Simulación del Acoplamiento Molecular , Péptido Sintasas/química , Péptido Sintasas/metabolismo , Uridina/químicaRESUMEN
DNA glycosylases are emerging as relevant pharmacological targets in inflammation, cancer and neurodegenerative diseases. Consequently, the search for inhibitors of these enzymes has become a very active research field. As a continuation of previous work that showed that 2-thioxanthine (2TX) is an irreversible inhibitor of zinc finger (ZnF)-containing Fpg/Nei DNA glycosylases, we designed and synthesized a mini-library of 2TX-derivatives (TXn) and evaluated their ability to inhibit Fpg/Nei enzymes. Among forty compounds, four TXn were better inhibitors than 2TX for Fpg. Unexpectedly, but very interestingly, two dithiolated derivatives more selectively and efficiently inhibit the zincless finger (ZnLF)-containing enzymes (human and mimivirus Neil1 DNA glycosylases hNeil1 and MvNei1, respectively). By combining chemistry, biochemistry, mass spectrometry, blind and flexible docking and X-ray structure analysis, we localized new TXn binding sites on Fpg/Nei enzymes. This endeavor allowed us to decipher at the atomic level the mode of action for the best TXn inhibitors on the ZnF-containing enzymes. We discovered an original inhibition mechanism for the ZnLF-containing Fpg/Nei DNA glycosylases by disulfide cyclic trimeric forms of dithiopurines. This work paves the way for the design and synthesis of a new structural class of inhibitors for selective pharmacological targeting of hNeil1 in cancer and neurodegenerative diseases.
Asunto(s)
ADN Glicosilasas/antagonistas & inhibidores , ADN Glicosilasas/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Purinas/química , Purinas/farmacología , Compuestos de Sulfhidrilo/química , Compuestos de Sulfhidrilo/farmacología , Bacterias/enzimología , Sitios de Unión , Cristalografía por Rayos X , Reparación del ADN , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Humanos , Modelos Moleculares , Conformación Molecular , Estructura Molecular , Unión Proteica , Relación Estructura-Actividad , Tioxantenos/química , Tioxantenos/farmacologíaRESUMEN
Nucleosides analogues are the cornerstone of the treatment of several human diseases. They are especially at the forefront of antiviral therapy. Their therapeutic efficiency depends on their capacity to be converted to the active nucleoside triphosphate form through successive phosphorylation steps catalyzed by nucleoside/nucleotide kinases. In this context, it is mandatory to develop a rapid, reliable and sensitive enzyme activity test to evaluate their metabolic pathways. In this study, we report a proof of concept to directly monitor on-line nucleotide multiple phosphorylation. The methodology was developed by on-line enzyme bioreactors hyphenated with High-Resolution Mass Spectrometry detection. Human Thymidylate Kinase (hTMPK) and human Nucleoside Diphosphate Kinase (hNDPK) were covalently immobilized on functionalized silica beads, and packed into micro-bioreactors (40⯵L). By continuous infusion of substrate into the bioreactors, the conversion of thymidine monophosphate (dTMP) into its di- (dTDP) and tri-phosphorylated (dTTP) forms was visualized by monitoring their Extracted Ion Chromatogram (EIC) of their [Mâ¯-â¯H]- ions. Both bioreactors were found to be robust and durable over 60 days (storage at 4⯰C in ammonium acetate buffer), after 20 uses and more than 750â¯min of reaction, making them suitable for routine analysis. Each on-line conversion step was shown rapid (<5â¯min), efficient (conversion efficiencyâ¯>â¯55%), precise and repeatable (CVâ¯<â¯3% for run-to-run analysis). The feasibility of the on-line multi-step conversion from dTMP to dTTP was also proved. In the context of selective antiviral therapy, this proof of concept was then applied to the monitoring of specificity of conversion of two synthesized Acyclic Nucleosides Phosphonates (ANPs), regarding human Thymidylate Kinase (hTMPK) and vaccina virus Thymidylate Kinase (vvTMPK).
Asunto(s)
Reactores Biológicos , Enzimas Inmovilizadas/química , Nucleósido-Fosfato Quinasa/química , Organofosfonatos/química , Timidina Monofosfato/química , Nucleótidos de Timina/química , Humanos , Espectrometría de Masas/métodos , Fosforilación , Prueba de Estudio Conceptual , Virus Vaccinia/enzimologíaRESUMEN
Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes is a series of Editorials which is published on a biannual basis by the Editorial Board of the Medicinal Chemistry section of the journal Molecules [...].
Asunto(s)
Química Farmacéutica/tendencias , Descubrimiento de Drogas/tendencias , HumanosRESUMEN
Herein, we propose the synthesis of a microspherical imprinted hydrogel meant for the controlled release of a nucleotide, adenosine 5'-monophosphate (5'-AMP). Indeed, molecularly imprinted polymers-based (MIPs) materials possess remarkable selective molecular recognition ability that mimicks biological systems. MIPs have been used in numerous applications and hold great promise for the vectorization and/or controlled release of therapeutics and cosmetics. But, the conception of imprinted hydrogels-based drug delivery systems that are able to release polar bioactive compounds is explored weakly. Herein, the synthesis of imprinted hydrogel microbeads by inverse Pickering emulsion is detailed. Microspheres showed a large 5'-AMP loading capacity, around 300â¯mg·g-1, and a high binding capacity comparatively to the non-imprinted counterpart. The MIP had a thermo-responsive release behavior providing sustained release of adenosine 5'-monophosphate in an aqueous buffer simulating both human skin pH and temperature.
Asunto(s)
Adenosina Monofosfato/administración & dosificación , Emulsiones/química , Hidrogeles/síntesis química , Microesferas , Impresión Molecular , Reactivos de Enlaces Cruzados/química , Preparaciones de Acción Retardada , Liberación de Fármacos , Cinética , Nanopartículas/química , Nanopartículas/ultraestructura , Tamaño de la Partícula , Polimerizacion , Dióxido de Silicio/química , Solventes , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Nucleosides and their analogues play a crucial role in the treatment of several diseases including cancers and viral infections. Their therapeutic efficiency depends on their capacity to be converted to the active nucleoside triphosphates form through successive phosphorylation steps catalyzed by nucleoside/nucleotide kinases. It is thus mandatory to develop an easy, rapid, reliable and sensitive enzyme activity tests. In this study, we monitored the three-step phosphorylation of thymidine to thymidine triphosphate respectively by (1) human thymidine kinase 1 (hTK1), (2) human thymidylate kinase (hTMPK) and (3) human nucleoside diphosphate kinase (hNDPK). Free and immobilized kinase activities were characterized by using the Michaelis-Menten kinetic model. Flow Injection Analysis (FIA) with High-Resolution Mass Spectrometry (HRMS) was used as well as capillary electrophoresis (CE) with UV detection. The three-step cascade phosphorylation of thymidine was also monitored. FIA-HRMS allows a sensitive and rapid evaluation of the phosphorylation process. This study proposes simple, rapid, efficient and sensitive methods for enzyme kinetic studies and successive phosphorylation monitoring with immobilized enzymes.
Asunto(s)
Enzimas Inmovilizadas/química , Nucleósido-Difosfato Quinasa/química , Nucleósido-Fosfato Quinasa/química , Timidina Quinasa/química , Timidina/química , Análisis de Inyección de Flujo/métodos , Humanos , Cinética , Espectrometría de Masas/métodos , Nanopartículas/química , FosforilaciónRESUMEN
Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes is a series of Editorials that is published on a biannual basis by the Editorial Board of the Medicinal Chemistry section of the journal Molecules [...].
Asunto(s)
Química Farmacéutica , Descubrimiento de Drogas , HumanosRESUMEN
Hitherto unknown chromophoric nucleosides are reported. This novel set of visibly coloured dye-labeled 5'-nucleosides, including 1,2,4,5-tetrazine, dicyanomethylene-4H-pyran, benzophenoxazinone, 9,10-anthraquinone and azobenzene chromophores, were prepared mainly under Cu-catalyzed azide-alkyne cycloaddition (CuAAC). The design criteria are outlined. Several derivatives possess in supplement a fluorescence property. The absorption and fluorescence spectra of all coloured nucleosides were recorded to study their potential as visible-range probes. Such nucleodyes are of great interest for future competitive lateral flow test MIP-based strips.