RESUMO
Harvesting sunlight to drive carbon dioxide (CO2) valorisation represents an ideal concept to support a sustainable and carbon-neutral economy. While the photochemical reduction of CO2 to carbon monoxide (CO) has emerged as a hot research topic, the full CO2-to-CO conversion remains an often-overlooked criterion that prevents a productive and direct valorisation of CO into high-value-added chemicals. Herein, we report a photocatalytic process that unlocks full and fast CO2-to-CO conversion (<10 min) and its straightforward valorisation into human health related field of radiochemistry with carbon isotopes. Guided by reaction-model-based kinetic simulations to rationalize reaction optimisations, this manifold opens new opportunities for the direct access to 11C- and 14C-labeled pharmaceuticals from their primary isotopic sources [11C]CO2 and [14C]CO2.
RESUMO
The need for carbon-labeled radiotracers is increasingly higher in drug discovery and development (carbon-14, ß-, t1/2 = 5730 years) as well as in positron emission tomography (PET) for in vivo molecular imaging applications (carbon-11, ß+, t1/2 = 20.4 min). However, the structural diversity of radiotracers is still systematically driven by the narrow available labeled sources and methodologies. In this context, the emergence of carbon dioxide radical anion chemistry might set forth potential unexplored opportunities. Based on a dynamic isotopic equilibration between formate salts and [13C, 14C, 11C]CO2, C-labeled radical anion CO2â¢- could be accessed under extremely mild conditions within seconds. This methodology was successfully applied to hydrocarboxylation and dicarboxylation reactions in late-stage carbon isotope labeling of pharmaceutically relevant compounds. The relevance of the method in applied radiochemistry was showcased by the whole-body PET biodistribution profile of [11C]oxaprozin in mice.
Assuntos
Dióxido de Carbono , Sais , Camundongos , Animais , Isótopos de Carbono , Radioisótopos de Carbono , Dióxido de Carbono/química , Distribuição Tecidual , Ânions , Tomografia por Emissão de Pósitrons/métodos , Formiatos , Marcação por IsótopoRESUMO
Carbon isotope labeling is a traceless technology, which allows tracking the fate of organic compounds either in the environment or in living organisms. This article reports on a general approach to label urea derivatives with all carbon isotopes, including 14C and 11C, based on a Staudinger aza-Wittig sequence. It provides access to all aliphatic/aromatic urea combinations.
Assuntos
Dióxido de Carbono/química , Radioisótopos de Carbono/química , Marcação por Isótopo/métodos , Ureia/químicaRESUMO
A general procedure for the late-stage [11C], [13C] and [14C]carbon isotope labeling of cyclic carbamates is reported. This protocol allows the incorporation of carbon dioxide, the primary source of carbon-14 and carbon-11 radioisotopes, in a direct, cost-effective and sustainable manner. A disconnection/reconnection strategy, involving ring opening/isotopic closure, was also implemented.
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A transition-metal-free carbon isotope exchange procedure on phenyl acetic acids is described. Utilizing the universal precursor CO2 , this protocol allows the carbon isotope to be inserted into the carboxylic acid position, with no need of precursor synthesis. This procedure enabled the labeling of 15 pharmaceuticals and was compatible with carbon isotopes [14 C] and [13 C]. A proof of concept with [11 C] was also obtained with low molar activity valuable for distribution studies.
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The understanding of the cellular uptake and the intracellular fate of nanoparticles and their subsequent influence on cell viability is challenging as far as micelles are concerned. Such systems are dynamic by nature, existing as unimers under their critical micelle concentration (CMC), and as micelles in equilibrium with unimers above the CMC, making canonical dose-response relationships difficult to establish. The purpose of this study was to investigate the in vitro cytotoxicity and uptake of two micellar sytems that are relevant for drug delivery. The two micelles incorporate a poly(ethylene glycol) coating and a pentacosadiynoic core which is either polymerized (pDA-PEG micelles) or non-polymerized (DA-PEG micelles), with the aim of evaluating the influence of the micelles status ("particle-like" or "dynamic", respectively) on their toxicological profile. Intracellular distribution and cytotoxicity of polymerized and non-polymerized micelles were investigated on RAW 264.7 macrophages in order to compare any different interactions with cells. Non-polymerized micelles showed significantly higher cytotoxicity than polymerized micelles, especially in terms of cell permeabilization, correlated to a higher accumulation in cell membranes. Other potential toxicity endpoints of polymerized micelles were then thoroughly studied in order to assess possible responses resulting from their endocytosis. No specific mechanisms of cytotoxicity were observed, neither in terms of apoptosis induction, cell membrane damage, release of inflammatory mediators nor genotoxicity. These data indicate that non-polymerized micelles accumulate in the cell membrane and induce cell membrane permeabilization, resulting in significant toxicity, whereas polymerized, stable micelles are internalized by cells but exert no or very low toxicity.
Assuntos
Micelas , Polímero Poliacetilênico/toxicidade , Animais , Apoptose , Portadores de Fármacos , Endocitose , Inflamação , L-Lactato Desidrogenase/metabolismo , Lipopolissacarídeos , Camundongos , Mitocôndrias/metabolismo , Nanopartículas/química , Nanopartículas/toxicidade , Nanoestruturas , Necrose , Permeabilidade , Polímero Poliacetilênico/química , Polietilenoglicóis/química , Polimerização , Células RAW 264.7RESUMO
A copper-catalyzed procedure enabling dynamic carbon isotope exchange is described. Utilizing the universal precursor [14C]CO2, this protocol allows to insert, in one single step, the desired carbon tag into carboxylic acids with no need of structural modifications. Reducing synthetic costs and limiting the generation of radioactive waste, this procedure will facilitate the access to carboxylic acids containing drugs and accelerate early 14C-based ADME studies supporting drug development.
Assuntos
Dióxido de Carbono/química , Ácidos Carboxílicos/química , Compostos Radiofarmacêuticos/química , Isótopos de Carbono/química , Radioisótopos de Carbono/química , Ácidos Carboxílicos/síntese química , Catálise , Cobre/química , Marcação por Isótopo/métodos , Compostos Radiofarmacêuticos/síntese químicaRESUMO
A robust, click-chemistry-inspired procedure for radiolabeling of cyclic ureas was developed. This protocol, suitable for all carbon isotopes (11 C, 13 C, 14 C), is based on the direct functionalization of carbon dioxide: the universal building block for carbon radiolabeling. The strategy is operationally simple and reproducible in different radiochemistry centers, exhibits remarkably wide substrate scope with short reaction times, and demonstrates superior reactivity as compared to previously reported systems. With this procedure, a variety of pharmaceuticals and an unprotected peptide were labeled with high radiochemical efficiency.
Assuntos
Dióxido de Carbono/química , Marcação por Isótopo , Compostos Radiofarmacêuticos/síntese química , Ureia/síntese química , Isótopos de Carbono , Química Click , Estrutura Molecular , Compostos Radiofarmacêuticos/química , Ureia/análogos & derivados , Ureia/químicaRESUMO
One-carbon metabolism is an ubiquitous metabolic pathway that encompasses the reactions transferring formyl-, hydroxymethyl- and methyl-groups bound to tetrahydrofolate for the synthesis of purine nucleotides, thymidylate, methionine and dehydropantoate, the precursor of coenzyme A. An alternative cyclic pathway was designed that substitutes 4-hydroxy-2-oxobutanoic acid (HOB), a compound absent from known metabolism, for the amino acids serine and glycine as one-carbon donors. It involves two novel reactions, the transamination of l-homoserine and the transfer of a one-carbon unit from HOB to tetrahydrofolate releasing pyruvate as coproduct. Since canonical reactions regenerate l-homoserine from pyruvate by carboxylation and subsequent reduction, every one-carbon moiety made available for anabolic reactions originates from CO2. The HOB-dependent pathway was established in an Escherichia coli auxotroph selected for prototrophy using long-term cultivation protocols. Genetic, metabolic and biochemical evidence support the emergence of a functional HOB-dependent one-carbon pathway achieved with the recruitment of the two enzymes l-homoserine transaminase and HOB-hydroxymethyltransferase and of HOB as an essential metabolic intermediate. Escherichia coli biochemical reprogramming was achieved by minimally altering canonical metabolism and leveraging on natural selection mechanisms, thereby launching the resulting strain on an evolutionary trajectory diverging from all known extant species.
Assuntos
Acetoacetatos/metabolismo , Carbono/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Melhoramento Genético/métodos , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/genética , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Glicina/genética , Glicina/metabolismo , Ácido Pirúvico/metabolismo , Serina/genética , Serina/metabolismo , Biologia Sintética/métodosRESUMO
The development of robust and straightforward methods to efficiently label aromatic moieties starting from simple and convenient radio-synthetic sources still represents a considerable challenge. In this report, a new palladium-catalyzed decarboxylative cyanation protocol has been described. This procedure utilizes [(14)C]-labeled potassium cyanide, one of the simplest and commercially available sources of carbon-14. Under the optimized reaction conditions, a series of [(13)C] and [(14)C]-aromatic nitriles were easily prepared (12-74% yield starting from potassium cyanide). The usefulness of this methodology is highlighted by a rare example of a formal two-step [(12)C]-[(14)C] carbon isotope exchange. The current synthetic approach may represent a promising alternative to traditional preparations of relevant building blocks such as labeled aromatic nitriles.
Assuntos
Radioisótopos de Carbono/química , Éteres de Hidroxibenzoatos/química , Paládio/química , Cianeto de Potássio/química , CatáliseRESUMO
Adenosine is a pleiotropic endogenous nucleoside with potential neuroprotective pharmacological activity. However, clinical use of adenosine is hampered by its extremely fast metabolization. To overcome this limitation, we recently developed a new squalenoyl nanomedicine of adenosine [Squalenoyl-Adenosine (SQAd)] by covalent linkage of this nucleoside to the squalene, a natural lipid. The resulting nanoassemblies (NAs) displayed a dramatic pharmacological activity both in cerebral ischemia and spinal cord injury pre-clinical models. The aim of the present study was to investigate the plasma profile and tissue distribution of SQAd NAs using both Squalenoyl-[(3)H]-Adenosine NAs and [(14)C]-Squalenoyl-Adenosine NAs as respective tracers of adenosine and squalene moieties of the SQAd bioconjugate. This study was completed by radio-HPLC analysis allowing to determine the metabolization profile of SQAd. We report here that SQAd NAs allowed a sustained circulation of adenosine under its prodrug form (SQAd) for at least 1h after intravenous administration, when free adenosine was metabolized within seconds after injection. Moreover, the squalenoylation of adenosine and its formulation as NAs also significantly modified biodistribution, as SQAd NAs were mainly captured by the liver and spleen, allowing a significant release of adenosine in the liver parenchyma. Altogether, these results suggest that SQAd NAs provided a reservoir of adenosine into the bloodstream which may explain the previously observed neuroprotective efficacy of SQAd NAs against cerebral ischemia and spinal cord injury.
Assuntos
Adenosina , Nanopartículas , Pró-Fármacos , Esqualeno , Adenosina/administração & dosagem , Adenosina/química , Adenosina/farmacocinética , Animais , Radioisótopos de Carbono , Cromatografia Líquida de Alta Pressão , Masculino , Camundongos , Nanopartículas/administração & dosagem , Nanopartículas/química , Pró-Fármacos/administração & dosagem , Pró-Fármacos/química , Pró-Fármacos/farmacocinética , Esqualeno/administração & dosagem , Esqualeno/química , Esqualeno/farmacocinética , Distribuição Tecidual , TrítioRESUMO
There is an urgent need to develop new therapeutic approaches for the treatment of severe neurological trauma, such as stroke and spinal cord injuries. However, many drugs with potential neuropharmacological activity, such as adenosine, are inefficient upon systemic administration because of their fast metabolization and rapid clearance from the bloodstream. Here, we show that conjugation of adenosine to the lipid squalene and the subsequent formation of nanoassemblies allows prolonged circulation of this nucleoside, providing neuroprotection in mouse stroke and rat spinal cord injury models. The animals receiving systemic administration of squalenoyl adenosine nanoassemblies showed a significant improvement of their neurologic deficit score in the case of cerebral ischaemia, and an early motor recovery of the hindlimbs in the case of spinal cord injury. Moreover, in vitro and in vivo studies demonstrated that the nanoassemblies were able to extend adenosine circulation and its interaction with the neurovascular unit. This Article shows, for the first time, that a hydrophilic and rapidly metabolized molecule such as adenosine may become pharmacologically efficient owing to a single conjugation with the lipid squalene.
RESUMO
A simple and efficient method for constructing 4-arylidene-5-imidazolones was developed using a phosphine-catalyzed tandem umpolung addition and intramolecular cyclization of amidine pronucleophiles on arylpropiolates. The reaction offers a robust route to heterocycle analogues of the fluorescent protein chromophores.
Assuntos
Imidazóis/síntese química , Fosfinas/química , Catálise , Ciclização , Imidazóis/química , Proteínas Luminescentes/química , Estrutura MolecularRESUMO
The development of new efficient syntheses of labeled reagents is a great challenge. Avoidance of overcomplicated procedures, availability and cost of starting materials are important considerations in choosing the synthetic route. In this report, we describe a facile and rapid preparation of labeled cyanate by ozonation of cyanide, a basic precursor. The crude cyanate was used without purification for the synthesis of various [(13)C] or [(14)C]ureidocarboxylic acids (20-68% yield from potassium cyanide). According to these results, cyanide ozonation may prove to be a promising alternative to traditional preparations of labeled cyanate.
Assuntos
Cianatos/síntese química , Marcação por Isótopo/métodos , Ozônio/química , Radioisótopos de Carbono/química , Ácidos Carboxílicos/química , Técnicas de Química Sintética/métodos , Oxirredução , Ureia/análogos & derivados , Ureia/químicaRESUMO
Conjugated linoleic acid (CLA) reduces adiposity in vivo. However, mechanisms mediating these changes are unclear. Therefore, we treated cultures of human adipocytes with trans-10, cis-12 (10,12) CLA, cis-9, trans-11 (9,11) CLA or other trans fatty acids (FA), and measured indices of lipid metabolism. The lipid-lowering effects of 10,12 CLA were unique, as other trans FA did not reduce TG content to the same extent. Using low levels of [(14)C]-CLA isomers, it was shown that both isomers were readily incorporated into acylglycerols and phospholipids, albeit at lower levels than [(14)C]-oleic or [(14)C]-linoleic acids. When using [(14)C]-acetic acid and [(14)C]-pyruvic acid as substrates, 30 µM 10,12 CLA, but not 9,11 CLA, decreased de novo synthesis of triglyceride, free FA, diacylglycerol, cholesterol esters, cardiolipin, phospholipids and ceramides within 3-24 h. Treatment with 30 µM 10,12 CLA, but not 9,11 CLA, decreased total cellular lipids within 3 days and the ratio of monounsaturated FA (MUFA) to saturated FA, and increased C18:0 acyl-CoA levels within 24 h. Consistent with these data, stearoyl-CoA desaturase (SCD)-1 mRNA and protein levels were down-regulated by 10,12 CLA within 7-12 h, respectively. The mRNA levels of liver X receptor (LXR)α and sterol regulatory element binding protein (SREBP)-1c, transcription factors that regulate SCD-1, were decreased by 10,12 CLA within 5 h. These data suggest that the isomer-specific decrease in de novo lipid synthesis by 10,12 CLA is due, in part, to the rapid repression of lipogenic transcription factors that regulate MUFA synthesis, suggesting an anti-obesity mechanism unique to this trans FA.
Assuntos
Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Ácidos Linoleicos Conjugados/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Adulto , Células Cultivadas , Ácidos Graxos/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Feminino , Humanos , Isomerismo , Receptores X do Fígado , Pessoa de Meia-Idade , Receptores Nucleares Órfãos/genética , Receptores Nucleares Órfãos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Estearoil-CoA Dessaturase/genética , Estearoil-CoA Dessaturase/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Ácidos Graxos trans/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Triglicerídeos/biossíntese , Adulto JovemRESUMO
Elaidic acid (trans-9-C18:1 or trans-9) is assumed to exert atherogenic effects due to its double bond configuration. The possibility that trans-9 and vaccenic acid (trans-11-C18:1 or trans-11), its positional isomer, were biochemically equivalent and interchangeable compounds, was investigated by reference to their cis-isomers through esterification-related activities using rat liver cells and subcellular fractions. In hepatocytes, both trans-C18:1 were incorporated to the same extent in triacylglycerols, but trans-9 was more esterified than trans-11 into phospholipids (P < 0.05). Glycerol-3-phosphate acyltransferase activity in microsomes was lower with trans-11 than with trans-9, while this activity in mitochondria was ~40% greater with trans-11 than with trans-9 (P < 0.05). Activity of 2-lysophosphatidic acid acyltransferase in microsomes was of comparable extent with both trans isomers, but activity of 2-lysophosphatidylcholine acyltransferase was significantly greater with trans-9 than with trans-11 at P < 0.01. Lipoproteins secreted by hepatocytes reached equivalent levels in the presence of any isomers, but triacylglycerol production was more elevated with trans-11 than with trans-9 at P < 0.05. Cholesterol efflux from previously labelled hepatocytes was lower with trans-11 than with trans-9. When these cells were exposed to either trans-C18:1, the gene expression of proteins involved in fatty acid esterification and lipoprotein synthesis was unaffected, which indicates that the biochemical differences essentially depended on enzyme/substrate affinities. On the whole, vaccenic and elaidic acid were shown to incorporate cell phospholipids unequally, at least in vitro, which suggests they can differently affect lipid metabolic pathways in normal cells.
Assuntos
Hepatócitos/metabolismo , Fígado/metabolismo , Ácido Oleico/farmacocinética , Ácidos Oleicos/farmacocinética , Fosfolipídeos/metabolismo , Triglicerídeos/metabolismo , Animais , Técnicas de Cultura de Células , Colesterol/metabolismo , Esterificação/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Glicerol-3-Fosfato O-Aciltransferase/efeitos dos fármacos , Glicerol-3-Fosfato O-Aciltransferase/metabolismo , Hepatócitos/efeitos dos fármacos , Isomerismo , Lipoproteínas/metabolismo , Fígado/efeitos dos fármacos , Masculino , Microssomos/efeitos dos fármacos , Microssomos/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Ácido Oleico/administração & dosagem , Ácido Oleico/química , Ácidos Oleicos/administração & dosagem , Ácidos Oleicos/química , Ratos , Ratos Wistar , Equivalência TerapêuticaRESUMO
Few studies report the individual effect of 9c,11t- and 10t,12c-CLA on human energy metabolism. We compared the postprandial oxidative metabolism of 9c,11t- and 10t,12c-CLA and oleic acid (9c-18:1) in 22 healthy moderately overweight volunteers. After 24 weeks supplementation with 9c,11t-, 10t,12c-CLA or 9c-18:1 (3 g/day), subjects consumed a single oral bolus of the appropriate [1-(13)C]-labeled fatty acid. 8 h post-dose, cumulative oxidation was similar for 9c-18:1 and 10t,12c (P = 0.66), but significantly higher for 9c,11t (P < 0.01).
Assuntos
Saúde , Ácido Oleico/metabolismo , Sobrepeso/metabolismo , Período Pós-Prandial , Adulto , Progressão da Doença , Método Duplo-Cego , Feminino , Humanos , Isomerismo , Ácidos Linoleicos Conjugados/administração & dosagem , Ácidos Linoleicos Conjugados/química , Ácidos Linoleicos Conjugados/metabolismo , Ácidos Linoleicos Conjugados/farmacocinética , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Ácido Oleico/administração & dosagem , Ácido Oleico/química , Ácido Oleico/farmacocinética , Oxirredução , PlacebosRESUMO
A new drug carrier system based on self-assembly and polymerization of polydiacetylenic amphiphiles is described. Although classical amphiphiles can help in solubilizing hydrophobic molecules upon self-arrangement into a variety of nanometric structures, a greater effect on drug loading was observed for our polymerized micelles as compared to the non-polymerized analogues. This permitted higher aqueous solubilization of lipophilic drugs with low micelle concentration. (14)C labeling of a model drug on one side and of the amphiphile on the other side permitted assessment, after intravenous injection, of biodistribution and excretion profiles of the drug cargo.
Assuntos
Micelas , Polímeros/química , Poli-Inos/química , Animais , Interações Hidrofóbicas e Hidrofílicas , Masculino , Camundongos , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Polímero Poliacetilênico , Polímeros/metabolismo , Poli-Inos/metabolismo , Ratos , Ratos Wistar , SolubilidadeRESUMO
Vaccenic acid (trans-11-C(18:1)) chemically resembles elaidic acid (trans-9-C(18:1)) which is assumed to increase the risk of cardiovascular diseases, and thus could exert similar effects. Possible different oxidation rates of vaccenic versus elaidic acid were checked in muscles and liver, and through related gene expression in normal rat liver cells. In hepatic mitochondria, carnitine palmitoyltransferase (CPT) I exhibited comparable activity rates with both trans-isomers. CPT II activity was 30% greater (P < 0.05) with vaccenic than with elaidic acid as nonesterified fatty acids (NEFAs) or acyl-CoAs. Activity of the first beta-oxidation step was similar between the isomers in all the tissue slices and liver extracts assayed. Respiration rates were comparable with both trans-isomers as NEFAs in various liver extracts, but were 30% greater (P < 0.05) with vaccenoyl-CoA than with elaidoyl-CoA in liver mitochondria. Vaccenic acid was oxidised 25% more (P < 0.05) by liver peroxisomes than elaidic acid. In hepatocytes cultured with trans- and corresponding cis-C(18:1) isomers, gene expression of CPT I, hydroxyacyl-CoA dehydrogenase and hydroxymethylglutaryl-CoA synthase was at least 100% increased (P < 0.05), but was unchanged with vaccenic acid, relative to controls. In conclusion, the position and geometry of the double bonds in acyl chains are suggested to confer on vaccenic and elaidic acid specific biochemical properties that might differently affect their fates in tissues.