Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
Más filtros












Base de datos
Intervalo de año de publicación
1.
Chimia (Aarau) ; 77(6): 376-383, 2023 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38047776

RESUMEN

The fortieth anniversary of biocatalysis started at Ciba-Geigy and later at Novartis is a great time to pause and reflect on development of science and technology in this field. Enzyme-based synthesis became a highly valued enabling tool for pharmaceutical research and development over the last decades. In this perspective we aim to discuss how the scientific approaches and trends evolved over the time and present future challenges and opportunities.


Asunto(s)
Biocatálisis
2.
Cell Chem Biol ; 28(10): 1407-1419.e6, 2021 10 21.
Artículo en Inglés | MEDLINE | ID: mdl-33794192

RESUMEN

Three limonoid natural products with selective anti-proliferative activity against BRAF(V600E) and NRAS(Q61K)-mutation-dependent melanoma cell lines were identified. Differential transcriptome analysis revealed dependency of compound activity on expression of the mitochondrial cytochrome P450 oxidase CYP27A1, a transcriptional target of melanogenesis-associated transcription factor (MITF). We determined that CYP27A1 activity is necessary for the generation of a reactive metabolite that proceeds to inhibit cellular proliferation. A genome-wide small interfering RNA screen in combination with chemical proteomics experiments revealed gene-drug functional epistasis, suggesting that these compounds target mitochondrial biogenesis and inhibit tumor bioenergetics through a covalent mechanism. Our work suggests a strategy for melanoma-specific targeting by exploiting the expression of MITF target gene CYP27A1 and inhibiting mitochondrial oxidative phosphorylation in BRAF mutant melanomas.


Asunto(s)
Colestanotriol 26-Monooxigenasa/metabolismo , Limoninas/farmacología , Mitocondrias/efectos de los fármacos , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Productos Biológicos/química , Productos Biológicos/metabolismo , Productos Biológicos/farmacología , Productos Biológicos/uso terapéutico , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Colestanotriol 26-Monooxigenasa/antagonistas & inhibidores , Colestanotriol 26-Monooxigenasa/genética , Humanos , Limoninas/química , Limoninas/metabolismo , Limoninas/uso terapéutico , Melanoma/tratamiento farmacológico , Melanoma/patología , Factor de Transcripción Asociado a Microftalmía/genética , Factor de Transcripción Asociado a Microftalmía/metabolismo , Mitocondrias/metabolismo , Fosforilación Oxidativa/efectos de los fármacos , Regiones Promotoras Genéticas , Unión Proteica , Proteínas Proto-Oncogénicas B-raf/antagonistas & inhibidores , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas B-raf/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/metabolismo
3.
J Med Chem ; 63(19): 10773-10781, 2020 10 08.
Artículo en Inglés | MEDLINE | ID: mdl-32667203

RESUMEN

Visceral leishmaniasis is responsible for up to 30,000 deaths every year. Current treatments have shortcomings that include toxicity and variable efficacy across endemic regions. Previously, we reported the discovery of GNF6702, a selective inhibitor of the kinetoplastid proteasome, which cleared parasites in murine models of leishmaniasis, Chagas disease, and human African trypanosomiasis. Here, we describe the discovery and characterization of LXE408, a structurally related kinetoplastid-selective proteasome inhibitor currently in Phase 1 human clinical trials. Furthermore, we present high-resolution cryo-EM structures of the Leishmania tarentolae proteasome in complex with LXE408, which provides a compelling explanation for the noncompetitive mode of binding of this novel class of inhibitors of the kinetoplastid proteasome.


Asunto(s)
Antiprotozoarios/química , Antiprotozoarios/farmacología , Leishmaniasis Visceral/tratamiento farmacológico , Oxazoles/química , Oxazoles/farmacología , Inhibidores de Proteasoma/química , Inhibidores de Proteasoma/farmacología , Pirimidinas/química , Pirimidinas/farmacología , Animales , Antiprotozoarios/uso terapéutico , Perros , Humanos , Leishmania donovani/efectos de los fármacos , Leishmania donovani/aislamiento & purificación , Leishmania major/efectos de los fármacos , Leishmania major/aislamiento & purificación , Leishmaniasis Visceral/parasitología , Hígado/parasitología , Macaca fascicularis , Ratones , Ratones Endogámicos BALB C , Oxazoles/uso terapéutico , Inhibidores de Proteasoma/uso terapéutico , Pirimidinas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Triazoles/química
4.
Xenobiotica ; 50(2): 150-169, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31006307

RESUMEN

Asciminib is a potent, specific BCR-ABL1 inhibitor being developed for the treatment of patients with chronic myelogenous leukemia (CML) and Philadelphia chromosome positive acute lymphoblastic leukemia (Ph + ALL).Here, we present the results of human oral absorption, distribution, metabolism, excretion (ADME) and in vitro studies that together provide an overall understanding of the metabolism, distribution and clearance of asciminib in humans.Asciminib was rapidly absorbed with a maximum plasma concentration at two hours post-dose. Total radioactivity and asciminib showed similar terminal half-lives in plasma.Oral asciminib absorption ranged between a minimum of 33%, and a maximum of 57% based on the metabolite profiles of late time-point feces collections.Asciminib was eliminated mainly through feces via unchanged asciminib excretion and metabolism.Direct glucuronidation and oxidation were major metabolic pathways in human that were catalyzed predominantly by UDP-glucuronosyltransferase (UGT)2B7 and cytochrome P450 (CYP)3A4, respectively.The relative contribution of the glucuronidation pathway to the total clearance of asciminib via metabolism is estimated to range ∼28-58%, whereas the relative contribution of the oxidative pathway is estimated to range ∼37-64%, based upon the maximum oral absorption in humans.


Asunto(s)
Niacinamida/análogos & derivados , Inhibidores de Proteínas Quinasas/metabolismo , Pirazoles/metabolismo , Adulto , Proteínas de Fusión bcr-abl/metabolismo , Humanos , Masculino , Niacinamida/metabolismo
5.
Biotechnol Bioeng ; 115(9): 2156-2166, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-29943426

RESUMEN

Cytochrome P450 mono-oxygenases (P450) are versatile enzymes which play essential roles in C-source assimilation, secondary metabolism, and in degradations of endo- and exogenous xenobiotics. In humans, several P450 isoforms constitute the largest part of phase I metabolizing enzymes and catalyze oxidation reactions which convert lipophilic xenobiotics, including drugs, to more water soluble species. Recombinant human P450s and microorganisms are applied in the pharmaceutical industry for the synthesis of drug metabolites for pharmacokinetics and toxicity studies. Compared to the membrane-bound eukaryotic P450s, prokaryotic ones exhibit some advantageous features, such as high stability and generally easier heterologous expression. Here, we describe a novel P450 from Streptomyces platensis DSM 40041 classified as CYP107L that efficiently converts several commercial drugs of various size and properties. This P450 was identified by screening of actinobacterial strains for amodiaquine and ritonavir metabolizing activities, followed by genome sequencing and expression of the annotated S. platensis P450s in Escherichia coli. Performance of CYP107L in biotransformations of amodiaquine, ritonavir, amitriptyline, and thioridazine resembles activities of the main human metabolizing P450s, namely CYPs 3A4, 2C8, 2C19, and 2D6. For application in the pharmaceutical industry, an E. coli whole-cell biocatalyst expressing CYP107L was developed and evaluated for preparative amodiaquine metabolite production.


Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Oxigenasas de Función Mixta/metabolismo , Streptomyces/enzimología , Xenobióticos/metabolismo , Amodiaquina/metabolismo , Antimaláricos/metabolismo , Antivirales/metabolismo , Biotransformación , Clonación Molecular , Sistema Enzimático del Citocromo P-450/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Humanos , Inactivación Metabólica , Oxigenasas de Función Mixta/genética , Ritonavir/metabolismo , Análisis de Secuencia de ADN , Streptomyces/genética
6.
Genome Announc ; 5(28)2017 Jul 13.
Artículo en Inglés | MEDLINE | ID: mdl-28705965

RESUMEN

The three Actinobacteria strains Streptomyces platensis DSM 40041, Pseudonocardia autotrophica DSM 535, and Streptomyces fradiae DSM 40063 were described to selectively oxyfunctionalize several drugs. Here, we present their draft genomes to unravel their gene sets encoding promising cytochrome P450 monooxygenases associated with the generation of drug metabolites.

7.
Drug Metab Dispos ; 45(7): 817-825, 2017 07.
Artículo en Inglés | MEDLINE | ID: mdl-28442499

RESUMEN

Fevipiprant is a novel oral prostaglandin D2 receptor 2 (DP2; also known as CRTh2) antagonist, which is currently in development for the treatment of severe asthma and atopic dermatitis. We investigated the absorption, distribution, metabolism, and excretion properties of fevipiprant in healthy subjects after a single 200-mg oral dose of [14C]-radiolabeled fevipiprant. Fevipiprant and metabolites were analyzed by liquid chromatography coupled to tandem mass spectrometry and radioactivity measurements, and mechanistic in vitro studies were performed to investigate clearance pathways and covalent plasma protein binding. Biotransformation of fevipiprant involved predominantly an inactive acyl glucuronide (AG) metabolite, which was detected in plasma and excreta, representing 28% of excreted drug-related material. The AG metabolite was found to covalently bind to human plasma proteins, likely albumin; however, in vitro covalent binding to liver protein was negligible. Excretion was predominantly as unchanged fevipiprant in urine and feces, indicating clearance by renal and possibly biliary excretion. Fevipiprant was found to be a substrate of transporters organic anion transporter 3 (OAT3; renal uptake), multidrug resistance gene 1 (MDR1; possible biliary excretion), and organic anion-transporting polypeptide 1B3 (OATP1B3; hepatic uptake). Elimination of fevipiprant occurs via glucuronidation by several uridine 5'-diphospho glucuronosyltransferase (UGT) enzymes as well as direct excretion. These parallel elimination pathways result in a low risk of major drug-drug interactions or pharmacogenetic/ethnic variability for this compound.


Asunto(s)
Hepatocitos/metabolismo , Ácidos Indolacéticos/farmacocinética , Microsomas Hepáticos/metabolismo , Piridinas/farmacocinética , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Administración Oral , Adolescente , Adulto , Biotransformación , Heces/química , Voluntarios Sanos , Humanos , Técnicas In Vitro , Ácidos Indolacéticos/sangre , Ácidos Indolacéticos/orina , Masculino , Tasa de Depuración Metabólica , Metaboloma , Persona de Mediana Edad , Unión Proteica , Piridinas/sangre , Piridinas/orina , Eliminación Renal , Distribución Tisular , Adulto Joven
8.
J Biotechnol ; 235: 3-10, 2016 Oct 10.
Artículo en Inglés | MEDLINE | ID: mdl-27021957

RESUMEN

Human xanthine oxidoreductase (XOR), which is responsible for the final steps of the purine metabolism pathway and involved in oxidative drug metabolism, was successfully expressed in Escherichia coli BL21(DE3) Gold. Recombinant human (rh) XOR yielded higher productivity with the gene sequence optimized for expression in E.coli than with the native gene sequence. Induction of XOR expression with lactose or IPTG resulted in complete loss of activity whereas shake flasks cultures using media rather poor in nutrients resulted in functional XOR expression in the stationary phase. LB medium was used for a 25L fermentation in fed-batch mode, which led to a 5 fold increase of the enzyme productivity when compared to cultivation in shake flasks. Quinazoline was used as a substrate on the semi-preparative scale using an optimized whole cell biotransformation protocol, yielding 73mg of the isolated product, 4-quinazolinone, from 104mg of starting material.


Asunto(s)
Escherichia coli/genética , Proteínas Recombinantes , Xantina Oxidasa , Biotecnología , Fermentación , Humanos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Xantina Oxidasa/química , Xantina Oxidasa/genética , Xantina Oxidasa/aislamiento & purificación , Xantina Oxidasa/metabolismo
9.
Drug Metab Dispos ; 44(5): 653-64, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26921386

RESUMEN

KAE609 [(1'R,3'S)-5,7'-dichloro-6'-fluoro-3'-methyl-2',3',4',9'-tetrahydrospiro[indoline-3,1'-pyridol[3,4-b]indol]-2-one] is a potent, fast-acting, schizonticidal agent being developed for the treatment of malaria. After oral dosing of KAE609 to rats and dogs, the major radioactive component in plasma was KAE609. An oxidative metabolite, M18, was the prominent metabolite in rat and dog plasma. KAE609 was well absorbed and extensively metabolized such that low levels of parent compound (≤11% of the dose) were detected in feces. The elimination of KAE609 and metabolites was primarily mediated via biliary pathways (≥93% of the dose) in the feces of rats and dogs. M37 and M23 were the major metabolites in rat and dog feces, respectively. Among the prominent metabolites of KAE609, the isobaric chemical species, M37, was observed, suggesting the involvement of an isomerization or rearrangement during biotransformation. Subsequent structural elucidation of M37 revealed that KAE609, a spiroindolone, undergoes an unusual C-C bond cleavage, followed by a 1,2-acyl shift to form a ring expansion metabolite M37. The in vitro metabolism of KAE609 in hepatocytes was investigated to understand this novel biotransformation. The metabolism of KAE609 was qualitatively similar across the species studied; thus, further investigation was conducted using human recombinant cytochrome P450 enzymes. The ring expansion reaction was found to be primarily catalyzed by cytochrome P450 (CYP) 3A4 yielding M37. M37 was subsequently oxidized to M18 by CYP3A4 and hydroxylated to M23 primarily by CYP1A2. Interestingly, M37 was colorless, whereas M18 and M23 showed orange yellow color. The source of the color of M18 and M23 was attributed to their extended conjugated system of double bonds in the structures.


Asunto(s)
Indoles/metabolismo , Indoles/farmacología , Malaria/tratamiento farmacológico , Compuestos de Espiro/metabolismo , Compuestos de Espiro/farmacología , Animales , Bilis/metabolismo , Biotransformación/efectos de los fármacos , Sistema Enzimático del Citocromo P-450/metabolismo , Perros , Heces/química , Hepatocitos/metabolismo , Humanos , Hidroxilación , Masculino , Ratas , Ratas Wistar
10.
Microb Cell Fact ; 14: 82, 2015 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-26062974

RESUMEN

BACKGROUND: Getting access to authentic human drug metabolites is an important issue during the drug discovery and development process. Employing recombinant microorganisms as whole-cell biocatalysts constitutes an elegant alternative to organic synthesis to produce these compounds. The present work aimed for the generation of an efficient whole-cell catalyst based on the flavin monooxygenase isoform 2 (FMO2), which is part of the human phase I metabolism. RESULTS: We show for the first time the functional expression of human FMO2 in E. coli. Truncations of the C-terminal membrane anchor region did not result in soluble FMO2 protein, but had a significant effect on levels of recombinant protein. The FMO2 biocatalysts were employed for substrate screening purposes, revealing trifluoperazine and propranolol as FMO2 substrates. Biomass cultivation on the 100 L scale afforded active catalyst for biotransformations on preparative scale. The whole-cell conversion of trifluoperazine resulted in perfectly selective oxidation to 48 mg (46% yield) of the corresponding N (1)-oxide with a purity >98%. CONCLUSIONS: The generated FMO2 whole-cell catalysts are not only useful as screening tool for human metabolites of drug molecules but more importantly also for their chemo- and regioselective preparation on the multi-milligram scale.


Asunto(s)
Escherichia coli/genética , Oxigenasas de Función Mixta/química , Oxigenasas de Función Mixta/metabolismo , Preparaciones Farmacéuticas/metabolismo , Biocatálisis , Dinitrocresoles/metabolismo , Escherichia coli/metabolismo , Expresión Génica , Humanos , Oxigenasas de Función Mixta/genética , Propranolol/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad por Sustrato , Trifluoperazina/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...