RESUMO
The interaction of active substances with molecular structures in stratum corneum (SC) is crucial for the efficacy and safety of cosmetic formulations and topical drugs. However, the molecular architecture of SC is highly complex and methods to unambiguously localize exogenous molecules within SC are lacking. Consequently, little is known about the distribution of actives within SC, and proposed penetration mechanisms through SC are typically limited to simple diffusion via a tortuous (lipid only) or transverse (across corneocytes and lipid matrix) pathway. In this work, 3D mass spectrometry imaging is used to determine the spatial distributions of four active substances at subcellular resolution in SC, including partitioning between the corneocytes and the intercellular lipid matrix. The results indicate that caffeine, 2-methyl resorcinol and oxybenzone are homogeneously distributed in the corneocytes but largely absent in the lipid matrix, despite considerable differences in lipophilicity. In contrast, the distribution- of jasmonic acid derivative is more inhomogeneous and indicates considerable localization to both the lipid phase and the corneocytes.
Assuntos
Epiderme , Epiderme/metabolismo , Lipídeos/química , Lipídeos/análise , Humanos , Cafeína/metabolismo , Animais , Benzofenonas/metabolismo , Resorcinóis/metabolismo , Resorcinóis/farmacologia , Espectrometria de MassasRESUMO
Aims: Epidemiological studies that use dietary biomarkers to investigate the association between whole grain intake and the risk of obesity are sparse. We assessed the association between urinary alkylresorcinol metabolites including 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA), biomarkers of whole grain wheat and rye intake, and body fat measures. Methods: We measured urinary excretion of DHPPA and DHBA, body weight, height, and circumferences of the waist and hip at the baseline and again after 1-year in a representative sample of 306 community-dwelling adults in Huoshan, China. We also measured liver fat accumulation [indicated by the controlled attenuation parameter (CAP)] and other body composition after 1 year. Multivariate-adjusted linear models and linear mixed-effects models were used to analyze single measurement and repeated measurements, respectively. Results: Each 1 µg g-1 creatinine increase in urinary DHPPA levels was associated with 0.21%, 0.23%, 3.64%, and 4.80% decrease in body weight, body mass index (BMI), body fat mass (BFM) and visceral fat level (VFL), respectively (all P < 0.05). Higher DHBA levels were inversely associated with CAP (percentage difference per 1 µg g-1 creatinine increment: -1.98%, P < 0.05). Higher total urinary alkylresorcinol metabolite (DHPPA + DHBA) levels were associated with lower body weight, BMI, BFM, VFL, and CAP, with the percentage differences per 1 µg g-1 creatinine increment of -0.27%, -0.27%, -3.79%, -5.12%, and -2.24%, respectively (all P < 0.05). Conclusions: Our findings suggest that the intake of whole grain wheat and rye, reflected by urinary DHPPA and DHBA, is favorably associated with liver fat and other fat measures.
Assuntos
Biomarcadores , Fígado , Resorcinóis , Secale , Triticum , Grãos Integrais , Humanos , Secale/metabolismo , Triticum/metabolismo , Masculino , Feminino , Biomarcadores/urina , Pessoa de Meia-Idade , Resorcinóis/urina , Resorcinóis/metabolismo , Adulto , Fígado/metabolismo , Índice de Massa Corporal , China , Tecido Adiposo/metabolismo , Hidroxibenzoatos/urina , Hidroxibenzoatos/metabolismo , Propionatos/urina , Propionatos/metabolismo , Obesidade/metabolismo , Obesidade/urina , FenilpropionatosRESUMO
BACKGROUND: Benvitimod (Tapinarof), as a small-molecule topical therapeutical aryl hydrocarbon receptor (AHR)-modulating agent, is in clinical development for treating psoriasis and atopic dermatitis. Benvitimod reduces proinflammatory cytokines in psoriasis by specifically binding and activation of AHR. However, whether benvitimod can inhibit keratinocyte proliferation remains unclear. Minichromosome maintenance protein 6 (MCM6) is a key element of the prereplication complex (pre-RC) assembly which is one of the essential steps in the initiation of DNA replication for cell proliferation. OBJECTIVES: This study aimed to determine whether benvitimod could reduce the excessive proliferation of psoriatic keratinocytes by inhibiting MCM6. METHODS: We examined the inhibitory effect of benvitimod on MCM6-mediated proliferation of keratinocytes by HaCaT cells in vitro and an IMQ-induced psoriatic model of mice in vivo. RESULTS: Epidermal MCM6 expression was enhanced in the skin lesions of psoriatic patients. The experiments further revealed that MCM6 was required for the proliferation of keratinocytes and governed by the IL-22/STAT3 pathway. In addition, the antiproliferation effect of benvitimod is achieved by the inhibition of p-JAK1 and p-JAK2, which further restrained the activation of STAT3 in keratinocytes. Lastly, benvitimod could repressed imiquimod-induced skin lesions and the expression of epidermal MCM6 and p-STAT3 in mice. Moreover, knockdown of AHR in keratinocytes enhanced the activation of JAK1 and JAK2. CONCLUSION: The findings reveal that benvitimod could decrease MCM6-mediated proliferation of keratinocytes by affecting the JAK/STAT3 pathway, thereby serving as a new treatment modality for psoriasis.
Assuntos
Queratinócitos , Psoríase , Animais , Camundongos , Proliferação de Células , Imiquimode/farmacologia , Queratinócitos/metabolismo , Psoríase/patologia , Resorcinóis/metabolismo , Resorcinóis/farmacologia , Resorcinóis/uso terapêuticoRESUMO
Cytochrome P450 enzymes (CYPs) are one of the most important classes of oxidative enzymes in the human body, carrying out metabolism of various exogenous and endogenous substrates. In order to expand the knowledge of these enzymes' specificity and to obtain new natural product derivatives, CYP4F11, a cytochrome P450 monooxygenase, was used in the biotransformation of dialkylresorcinols 1 and 2, a pair of antibiotic microbial natural products. This investigation resulted in four biotransformation products including two oxidative products: a hydroxylated derivative (3) and a carboxylic acid derivative (4). In addition, acetylated (5) and esterified products (6) were isolated, formed by further metabolism by endogenous yeast enzymes. Oxidative transformations were highly regioselective, and took place exclusively at the ω-position of the C-5 alkyl chain. Homology modeling studies revealed that optimal hydrogen bonding between 2 and the enzyme can only be established with the C-5 alkyl chain pointing towards the heme. The closely-related CYP4F12 was not capable of oxidizing the dialkylresorcinol 2. Modeling experiments rationalize these differences by the different shapes of the binding pockets with respect to the non-oxidized alkyl chain. Antimicrobial testing indicated that the presence of polar groups on the side-chains reduces the antibiotic activity of the dialkylresorcinols.
Assuntos
Antibacterianos , Sistema Enzimático do Citocromo P-450 , Resorcinóis , Humanos , Antibacterianos/metabolismo , Biotransformação , Sistema Enzimático do Citocromo P-450/metabolismo , Oxirredução , Resorcinóis/metabolismoRESUMO
The interactions of two conformers of newly synthesized photoswitchable azobenzene analogue of methotrexate, called Phototrexate, with two cavitand derivatives, have been investigated in dimethyl sulfoxide medium. Photoluminescence methods have been applied to determine the complex stabilities and the related enthalpy and entropy changes associated to the complex formation around room temperature. Results show opposite temperature dependence of complex stabilities. The structure of the upper rims of the host molecules and the reordered solvent structure were identified as the background of the opposite tendencies of temperature dependence at molecular level. These results can support the therapeutic application of the photoswitchable phototrexate, because the formation of inclusion complexes is a promising method to regulate the pharmacokinetics of drug molecules.
Assuntos
Compostos Azo/química , Éteres Cíclicos/química , Metotrexato/química , Resorcinóis/química , Compostos Azo/metabolismo , Éteres Cíclicos/metabolismo , Isomerismo , Metotrexato/metabolismo , Modelos Moleculares , Estrutura Molecular , Resorcinóis/metabolismo , Temperatura , TermodinâmicaRESUMO
Most small molecules interact with several target proteins but this polypharmacology is seldom comprehensively investigated or explicitly exploited during drug discovery. Here, we use computational and experimental methods to identify and systematically characterize the kinase cross-pharmacology of representative HSP90 inhibitors. We demonstrate that the resorcinol clinical candidates ganetespib and, to a lesser extent, luminespib, display unique off-target kinase pharmacology as compared with other HSP90 inhibitors. We also demonstrate that polypharmacology evolved during the optimization to discover luminespib and that the hit, leads, and clinical candidate all have different polypharmacological profiles. We therefore recommend the computational and experimental characterization of polypharmacology earlier in drug discovery projects to unlock new multi-target drug design opportunities.
Assuntos
Descoberta de Drogas , Evolução Molecular , Proteínas de Choque Térmico HSP90/metabolismo , Inibidores de Proteínas Quinases/metabolismo , Sítios de Ligação , Receptor com Domínio Discoidina 1/antagonistas & inibidores , Receptor com Domínio Discoidina 1/metabolismo , Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Humanos , Isoxazóis/química , Isoxazóis/metabolismo , Simulação de Acoplamento Molecular , Inibidores de Proteínas Quinases/química , Proteínas Proto-Oncogênicas c-abl/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-abl/metabolismo , Resorcinóis/química , Resorcinóis/metabolismo , Triazóis/química , Triazóis/metabolismoRESUMO
Antibiotic resistance and emerging viral pandemics have posed an urgent need for new anti-infective drugs. By screening our microbial extract library against the main protease of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the notorious ESKAPE pathogens, an active fraction was identified and purified, leading to an initial isolation of adipostatins A (1) and B (2). In order to diversify the chemical structures of adipostatins toward enhanced biological activities, a type III polyketide synthase was identified from the native producer, Streptomyces davawensis DSM101723, and was subsequently expressed in an E. coli host, resulting in the isolation of nine additional adipostatins 3-11, including two new analogs (9 and 11). The structures of 1-11 were established by HRMS, NMR, and chemical derivatization, including using a microgram-scale meta-chloroperoxybenzoic acid epoxidation-MS/MS analysis to unambiguously determine the double bond position in the alkyl chain. The present study discovered SARS-CoV-2 main protease inhibitory activity for the class of adipostatins for the first time. Several of the adipostatins isolated also exhibited antimicrobial activity against selected ESKAPE pathogens.
Assuntos
Aciltransferases/metabolismo , Anti-Infecciosos/química , Proteínas de Bactérias/metabolismo , Resorcinóis/química , Aciltransferases/antagonistas & inibidores , Aciltransferases/classificação , Aciltransferases/genética , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/classificação , Proteínas de Bactérias/genética , COVID-19/patologia , COVID-19/virologia , Proteases 3C de Coronavírus/antagonistas & inibidores , Proteases 3C de Coronavírus/metabolismo , Avaliação Pré-Clínica de Medicamentos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Conformação Molecular , Filogenia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Resorcinóis/isolamento & purificação , Resorcinóis/metabolismo , Resorcinóis/farmacologia , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/metabolismo , Streptomyces/enzimologia , Espectrometria de Massas em TandemRESUMO
For seven days of wheat growth, caryopsis remained the main source of 5-n-alkylresorcinols with C19 and/or C21 homolog as a main compound. Shoot contained small amount of these phenolic lipids; their average content was 3.23% of level obtained in the whole seedling. Moreover, 41.38% of resorcinolic lipids of seven-day-old shoot was accumulated in part of leaf covered by coleoptile. Interestingly, a removal of 1.07% of the primary pool of kernel alkylresorcinols by short-term washing (10 s) of wheat seed with acetone before planting decreased their level only in seed of seven-day old seedling. Compared to the respective controls, this treatment did not affect the amount of these lipids in the green part of seedling that proved that de novo synthesis of 5-n-alkylresorcinols takes place in shoots. The very similar homolog profiles of these lipids in four- and seven-day-old shoots turned out to be markedly less diversified than those found in respective seed samples. Compared to the mature wheat caryopsis, the rise in the content of very-long-chain homologs was observed only in the oldest shoot. Their increased accumulation was probably connected with formation of cuticular layer providing the defensive barrier against various phytopathogens.
Assuntos
Resorcinóis/metabolismo , Triticum/metabolismo , Germinação , Metabolismo dos Lipídeos , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Triticum/crescimento & desenvolvimentoRESUMO
Toward a sustainable synthesis of value-added chemicals, the method of CO2 utilization attracts great interest in chemical process engineering. Biotechnological CO2 fixation is a promising technology; however, efficient methods that can fix carbon dioxide are still limited. Instead, some parts of microbial decarboxylases allow the introduction of carboxy group into phenolic compounds using bicarbonate ion as a C1 building block. Here, we identified a unique decarboxylase from Arthrobacter sp. K8 that acts on resorcinol derivatives. A high-throughput colorimetric decarboxylase assay facilitated gene cloning of orsellinic acid decarboxylase from genomic DNA library of strain K8. Sequence analysis revealed that the orsellinic acid decarboxylase belonged to amidohydrolase 2 family, but shared low amino acid sequence identity with those of related decarboxylases. Enzymatic characterization unveiled that the decarboxylase introduces a carboxy group in a highly regio-selective manner. We applied the decarboxylase to enzymatic carboxylation of resorcinol derivatives. Using Escherichia coli expressing the decarboxylase gene as a whole cell biocatalyst, orsellinic acid, 2,4-dihydroxybenzoic acid, and 4-methoxysalicylic acid were produced in the presence of saturated bicarbonate. These findings could provide new insights into the production of useful phenolic acids from resorcinol derivatives.
Assuntos
Arthrobacter/enzimologia , Arthrobacter/genética , Carboxiliases/química , Carboxiliases/genética , Clonagem Molecular , Resorcinóis/química , Resorcinóis/metabolismo , Sequência de Aminoácidos , Arthrobacter/isolamento & purificação , Escherichia coli/genética , Hidroxibenzoatos , Cinética , Fenóis/metabolismo , Análise de Sequência , Solo , Microbiologia do Solo , Especificidade por SubstratoRESUMO
Wheat bran was solid state fermented by Fomitopsis pinicola. The results showed that the processing properties were increased by fermentation and the content of total phenol and alkylresorcinols was 5.91 and 1.55 times of the unfermented bran respectively by the 6th day. The total antioxidant capacity was 5.73 times of the unfermented sample by the 4th day. Electronic nose analysis showed that the fermented wheat bran had a special flavor. GC-MS analysis found that 4-ethyl-2-methoxy-phenol was the main flavor substance, which was sharply increased during the fermentation. Furthermore, the textural properties of the dough and bread containing fermented bran were significantly improved. The content of phytic acid in the bread was significantly decreased, while the protein, total phenol and alkylresorcinols contents were significantly increased. Results suggest that solid state fermentation by Fomitopsis pinicola is a promising way to improve wheat bran to a nutritious and flavorful cereal food ingredient.
Assuntos
Coriolaceae/metabolismo , Fibras na Dieta/análise , Antioxidantes/química , Técnicas de Cultura Celular por Lotes , Pão/análise , Nariz Eletrônico , Aromatizantes/análise , Cromatografia Gasosa-Espectrometria de Massas , Fenóis/química , Fenóis/metabolismo , Resorcinóis/química , Resorcinóis/metabolismo , Compostos Orgânicos Voláteis/análiseRESUMO
Sporormielones A-E (1-5), novel C-C coupled orsellinic acid derivative dimers containing tricyclic cores with a dimethylcyclopentenone unit, were obtained, of which 1-3 and 5 showed obvious short-term memory improvement activity in AD flies. Based on transcriptome analysis, 13C labelling, and gene deletion, their plausible biosynthetic mechanism was proposed.
Assuntos
Fármacos Neuroprotetores , Resorcinóis , Doença de Alzheimer , Animais , Ascomicetos/metabolismo , Dimerização , Dípteros , Perfilação da Expressão Gênica , Memória de Curto Prazo/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/metabolismo , Fármacos Neuroprotetores/farmacologia , Resorcinóis/química , Resorcinóis/metabolismo , Resorcinóis/farmacologiaRESUMO
Tapinarof is a stilbene drug that is used to treat psoriasis and atopic dermatitis, and is thought to function through regulation of the AhR and Nrf2 signaling pathways, which have also been linked to inflammatory bowel diseases. It is produced by the gammaproteobacterial Photorhabdus genus, which thus represents a model to probe tapinarof structural and functional transformations. We show that Photorhabdus transforms tapinarof into novel drug metabolism products that kill inflammatory bacteria, and that a cupin enzyme contributes to the conversion of tapinarof and related dietary stilbenes into novel dimers. One dimer has activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE), and another undergoes spontaneous cyclizations to a cyclopropane-bridge-containing hexacyclic framework that exhibits activity against Mycobacterium. These dimers lack efficacy in a colitis mouse model, whereas the monomer reduces disease symptoms.
Assuntos
Antibacterianos/metabolismo , Autoimunidade/efeitos dos fármacos , Fatores Imunológicos/metabolismo , Photorhabdus/metabolismo , Resorcinóis/metabolismo , Estilbenos/metabolismo , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Biotransformação , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Camundongos , Resorcinóis/química , Resorcinóis/farmacologia , Estilbenos/química , Estilbenos/farmacologiaRESUMO
Pseudomonas chlororaphis isolates have been studied intensively for their beneficial traits. P. chlororaphis species function as probiotics in plants and fish, offering plants protection against microbes, nematodes and insects. In this review, we discuss the classification of P. chlororaphis isolates within four subspecies; the shared traits include the production of coloured antimicrobial phenazines, high sequence identity between housekeeping genes and similar cellular fatty acid composition. The direct antimicrobial, insecticidal and nematocidal effects of P. chlororaphis isolates are correlated with known metabolites. Other metabolites prime the plants for stress tolerance and participate in microbial cell signalling events and biofilm formation among other things. Formulations of P. chlororaphis isolates and their metabolites are currently being commercialized for agricultural use.
Assuntos
Anti-Infecciosos/metabolismo , Biofilmes/crescimento & desenvolvimento , Fenazinas/metabolismo , Plantas/microbiologia , Probióticos , Pseudomonas chlororaphis/classificação , Acil-Butirolactonas/metabolismo , Agricultura , Antinematódeos/metabolismo , Cianeto de Hidrogênio/metabolismo , Inseticidas/metabolismo , Fenótipo , Plantas/imunologia , Pseudomonas chlororaphis/química , Pseudomonas chlororaphis/crescimento & desenvolvimento , Pseudomonas chlororaphis/fisiologia , Pirrolnitrina/metabolismo , Resorcinóis/metabolismo , Sideróforos/metabolismo , Compostos Orgânicos Voláteis/metabolismoRESUMO
Analyte-responsive chromo-fluorogenic reactions under accessible conditions are important for designing small-molecule spectroscopic probes. We describe a series of newly constructed motifs based on the chromo-fluorogenic reaction between catechol derivatives (typically hydroxytyrosol, dopamine, and levodopa) and naphthoresorcin (NR) in aqueous solution under ambient conditions. The weakly absorptive and fluorogenic catechols/NR was converted to products having visible absorption and bright fluorescence within several minutes. The chromo-fluorophores produced from this reaction had a maximum absorbance at 458 nm and emission at 480 nm with high fluorescence quantum yields (30-84%). Inspired by the tyrosinase-catalyzed hydroxylation of monophenols to catechols, the tyrosinase-enabled chromo-fluorogenic reaction was verified by using monophenol (typically tyrosol) as the substrate. In this regard, a dual-readout tyrosinase activity assay was developed by virtue of the in situ "turn-on" optical signals. Furthermore, a test of tyrosinase inhibition, by using a common inhibitor kojic acid, demonstrated the potential of the chromo-fluorogenic reaction for developing other tyrosinase related assays and signal transduction.
Assuntos
Catecóis/metabolismo , Corantes Fluorescentes/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Naftalenos/metabolismo , Resorcinóis/metabolismo , Agaricales/enzimologia , Catecóis/química , Corantes Fluorescentes/química , Estrutura Molecular , Monofenol Mono-Oxigenase/análise , Naftalenos/química , Resorcinóis/químicaRESUMO
Protein kinases play an indispensable role in signaling pathways that regulate tumor cell functions, which represent potent therapeutic targets in cancers. Dual-specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A) as a serine/threonine kinase has recently been reported to be upregulated in pancreatic ductal adenocarcinoma (PDAC) and show protumorigenic effect. By activity-guided phytochemical investigation of the extracts from Glycyrrhiza uralensis Fisch, we expect to find the effective constituents that can suppress pancreatic cancer cell proliferation and/or induce cells apoptotic by inhibiting DYRK1A. Eight isopentenyl-substituted compounds (1-8), including four coumarins (1-4), one benzofuran (5), and three flavonoids (6-8), were isolated and identified from G. uralensis Fisch. Among them, licocoumarone (LC, 5) showed effective inhibitory activity against DYRK1A with an IC50 value of 12.56 µM. Molecular docking analysis suggested that LC completely occupied the whole pocket of DYRK1A and formed obvious hydrophobic interactions and hydrogen bonds with DYRK1A residues. Further in vitro validation, including Microscale Thermophoresis (MST) and drug affinity responsive target stability (DARTS) techniques, demonstrated the specific combining capacity of LC to DYRK1A. Meanwhile, LC induced significant cytotoxicity against DYRK1A-overexpressing BxPC-3 cells with an IC50 value of 50.77 µM. Mechanism studies revealed that LC reduced c-MET protein level by inhibiting DYRK1A. These findings provide preliminary evidences that LC as a natural DYRK1A inhibitor suppresses human pancreatic adenocarcinoma BxPC-3 cell proliferation and induces cell apoptotic, which might present new options and possibilities for targeted therapies in pancreatic cancer therapy.
Assuntos
Apoptose/efeitos dos fármacos , Benzofuranos/farmacologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Resorcinóis/farmacologia , Benzofuranos/química , Benzofuranos/metabolismo , Sítios de Ligação , Domínio Catalítico , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Glycyrrhiza/química , Glycyrrhiza/metabolismo , Humanos , Ligação de Hidrogênio , Simulação de Acoplamento Molecular , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismo , Resorcinóis/química , Resorcinóis/metabolismo , Quinases Dyrk , Neoplasias PancreáticasRESUMO
Aims: Nonsteroidal anti-inflammatory drugs (NSAIDs), among the most commonly used drugs worldwide, are associated with gastrointestinal (GI) complications that severely limit the clinical utility of this essential class of pain medications. Here, we mechanistically dissect the protective impact of a natural product, malabaricone C (Mal C), on NSAID-induced gastropathy. Results: Mal C dose dependently diminished erosion of the stomach lining and inflammation in mice treated with NSAIDs with the protective impact translating to improvement in survival. By decreasing oxidative and nitrative stress, Mal C treatment prevented NSAID-induced mitochondrial dysfunction and cell death; nuclear factor κ-light-chain enhancer of activated B cell induction, release of proinflammatory cytokines and neutrophil infiltration; and disruptions in the vascular endothelial growth factor/endostatin balance that contributes to mucosal autohealing. Importantly, Mal C failed to impact the therapeutic anti-inflammatory properties of multiple NSAIDs in a model of acute inflammation. In all assays tested, Mal C proved as or more efficacious than the current first-line therapy for NSAID-dependent GI complications, the proton pump inhibitor omeprazole. Innovation: Given that omeprazole-mediated prophylaxis is, itself, associated with a shift in NSAID-driven GI complications from the upper GI to the lower GI system, there is a clear and present need for novel therapeutics aimed at ameliorating NSAID-induced gastropathy. Mal C provided significant protection against NSAID-induced gastric ulcerations impacting multiple critical signaling cascades contributing to inflammation, cell loss, extracellular matrix degradation, and angiogenic autohealing. Conclusion: Thus, Mal C represents a viable lead compound for the development of novel gastroprotective agents.
Assuntos
Indometacina/antagonistas & inibidores , Inflamação/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Substâncias Protetoras/farmacologia , Resorcinóis/farmacologia , Úlcera Gástrica/tratamento farmacológico , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Morte Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Indometacina/administração & dosagem , Inflamação/induzido quimicamente , Inflamação/patologia , Masculino , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Estrutura Molecular , Neovascularização Patológica/induzido quimicamente , Neovascularização Patológica/patologia , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/metabolismo , Resorcinóis/administração & dosagem , Resorcinóis/metabolismo , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/patologia , Cicatrização/efeitos dos fármacosRESUMO
An understanding of the bioavailability of topically applied cosmetics ingredients is key to predicting their local skin and systemic toxicity and making a safety assessment. We investigated whether short-term incubations with S9 from the reconstructed epidermal skin model, EpiSkin™, would give an indication of the rate of chemical metabolism and produce similar metabolites to those formed in incubations with human skin explants. Both have advantages: EpiSkin™ S9 is a higher-throughput assay, while the human skin explant model represents a longer incubation duration (24 hours) model integrating cutaneous distribution with metabolite formation. Here, we compared the metabolism of 10 chemicals (caffeine, vanillin, cinnamyl alcohol, propylparaben, 4-amino-3-nitrophenol, resorcinol, 4-chloroaniline, 2-amino-3-methyl-3H-imidazo[4,5-F]quinoline and 2-acetyl aminofluorene) in both models. Both models were shown to have functional Phase 1 and 2 enzymes, including cytochrome P450 activities. There was a good concordance between the models with respect to the level of metabolism (stable vs. slowly vs. extensively metabolized chemicals) and major early metabolites produced for eight chemicals. Discordant results for two chemicals were attributed to a lack of the appropriate cofactor (NADP+ ) in S9 incubations (cinnamyl alcohol) and protein binding influencing chemical uptake in skin explants (4-chloroaniline). These data support the use of EpiSkin™ S9 as a screening assay to provide an initial indication of the metabolic stability of a chemical applied topically. If required, chemicals that are not metabolized by EpiSkin™ S9 can be tested in longer-term incubations with in vitro human explant skin to determine whether it is slowly metabolized or not metabolized at all.
Assuntos
Células Cultivadas/efeitos dos fármacos , Cosméticos/metabolismo , Cosméticos/toxicidade , Testes de Irritação da Pele/métodos , Pele/efeitos dos fármacos , Acetofenonas/metabolismo , Acetofenonas/toxicidade , Compostos de Anilina/metabolismo , Compostos de Anilina/toxicidade , Animais , Benzaldeídos/metabolismo , Benzaldeídos/toxicidade , Benzilaminas/metabolismo , Benzilaminas/toxicidade , Cafeína/metabolismo , Humanos , Parabenos/metabolismo , Parabenos/toxicidade , Ácidos Pentanoicos/metabolismo , Ácidos Pentanoicos/toxicidade , Propanóis/metabolismo , Propanóis/toxicidade , Resorcinóis/metabolismo , Resorcinóis/toxicidadeRESUMO
Alzheimer´s disease (AD) is the most common form of dementia involving Aß and tau protein. So far, AD cure remains elusive, but considering that AD progresses throughout tau pathology, which turns tau protein an appropriate target, besides tau is also included in other neurodegenerative disorders named as tauopathies. Here, we have isolated seventeen compounds belonging to six lichens species. Due to scarce of spectroscopic data of the compound 5,7-dihydroxy-6-methylphthalide, we explained their structural elucidation based on NMR data. In this study, we show that only tenuiorin from Umbilicaria antarctica inhibited 50% of tau 4R at 100 µM. Then, we shown that molecular interactions of tenuiorin with the steric zipper model of the hexapeptide 306VQIVYK311 were studied by docking calculations and the results suggested that tenuiorin forms both hydrogen bonds with lysine and glutamine side chains and forms several hydrophobic interactions with valine and lysine from 306VQIVYK311 motif.
Assuntos
Ascomicetos/química , Depsídeos/isolamento & purificação , Líquens/química , Resorcinóis/isolamento & purificação , Proteínas tau/antagonistas & inibidores , Doença de Alzheimer/tratamento farmacológico , Regiões Antárticas , Ascomicetos/metabolismo , Sítios de Ligação , Depsídeos/química , Humanos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Ligação Proteica , Resorcinóis/química , Resorcinóis/metabolismo , Proteínas tau/metabolismoRESUMO
In the native pathway to therapeutic cannabinoid biosynthesis in Cannabis sativa, the three-step production of a key intermediate, olivetolic acid, is catalysed by the enzymes tetraketide synthase (TKS; linear tetraketide intermediate production in two stages) and olivetolic acid cyclase (OAC; final C2 â C7 aldol condensation). In the absence of OAC, a nonenzymatic C2 â C7 decarboxylative aldol condensation of the tetraketide intermediate occurs forming olivetol. TKS is a type III polyketide synthase, and the question arises why it is unable to form olivetolic acid directly, but instead forms this unwanted side product. We determined the TKS, CoA complex structure, and performed structurally guided mutagenesis studies to identify potential residues responsible for cyclization pathway discrimination in type III polyketide synthases. Prior studies suggested an 'aldol switch' is necessary to allow linear tetraketide intermediate release prior to cyclization, thereby enabling subsequent olivetolic acid production by OAC. However, our studies do not support the presence of a universal or predictable 'aldol switch' consensus sequence. Instead, we propose the mode of ordered active site water activation between type III polyketide synthases catalysing different cyclization mechanisms is subtle and homologue-specific. Our work indicates that subtle structural variations between homologous enzymes can have a major mechanistic impact on the catalytic outcome. This highlights the importance of embedding high-resolution structural analysis of multiple enzyme homologues with classical site-directed mutagenesis studies when investigating highly similar enzymes with different mechanistic pathway outcomes. ENZYMES: TKS, EC 2.3.1.206; OAC, EC 4.4.1.26; chalcone synthase, EC 2.3.1.74; stilbene synthase, EC 2.3.1.95; 2-PS, EC 2.3.1.-. ACCESSION NUMBERS: The atomic coordinates and structure factors for the crystal structure of TKS have been deposited in the Protein Data Bank with accession number 6GW3.
Assuntos
Cannabis/enzimologia , Policetídeo Sintases/metabolismo , Resorcinóis/metabolismo , Ciclização , Modelos Moleculares , Policetídeo Sintases/química , Conformação ProteicaRESUMO
Azotobacter vineladii is a Gram-negative bacterium that produces alginate and poly-hydroxybutyrate (PHB), two polymers of biotechnological interest. This bacterium has the ability to form desiccation-resistant cysts. In the cyst the membrane phospholipids are replaced with a family of phenolic lipids called alkylresorcinols (ARs). The alginate, PHB, and ARs are controlled by the GacS/A two-component system and the small regulatory RNA (sRNA) RsmZ1, belonging to the Rsm (Csr) regulatory system. The Rsm (Csr) systems usually possess two or more sRNAs, in this regard A. vinelandii is the bacterium with the highest number of rsm-sRNAs. Originally, the presence of two sRNAs of the RsmY family (RsmY1 and RsmY2) was reported, but in a subsequent work it was suggested that they conformed to a single sRNA. In this work we provide genetic evidence confirming that rsmY1 and rsmY2 constitute a single gene. Also, it was established that rsmY mutation decreased alginate and ARs production, but did not affect the PHB synthesis. Transcriptional studies showed that rsmY has its higher expression during the stationary growth phase, and in the absence of RsmZ1, rsmY increases its transcription. Interestingly, rsmY expression was influenced by the carbon source, but its expression did not correlate with alginate production.