In vitro metabolism studies of natural products are crucial in the early development of medicines

Moreira, Fernanda de Lima; Marques, Lucas Maciel Mauriz; Lopes, Norberto Peporine; Oliveira, Anderson Rodrigo Moraes de

doi:10.1007/s11101-016-9482-0

Cited by 6 publications

(6 citation statements)

References 140 publications

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“…In vitro metabolism assays are a valuable tool to assess drug metabolism properties in the primary stages of drug discovery and development. Liver microsomes pooled from animals or humans are the main in vitro model used for metabolite profiling due to ethical and economic reasons, reproducibility of the results, and the ability to compare data from different species [25,26]. Considering the potential use of CAS X as an antitumor agent, the aim of this work was to characterize the metabolism of CAS X using the in vitro microsomal model.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Casearin X Metabolism by Rat and Human Liver Microsomes

et al. 2018

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Casearin X (CAS X) is the major clerodane diterpene isolated from the leaves of Casearia sylvestris and has been extensively studied due to its powerful cytotoxic activity at low concentrations. Promising results for in vivo antitumor action have also been described when CAS X was administered intraperitoneally in mice. Conversely, loss of activity was observed when orally administered. Since the advancement of natural products as drug candidates requires satisfactory bioavailability for their pharmacological effect, this work aimed to characterize the CAS X metabolism by employing an in vitro microsomal model for the prediction of preclinical pharmacokinetic data. Rat and human liver microsomes were used to assess species differences. A high-performance liquid chromatography with diode-array detection (HPLC-DAD) method for the quantification of CAS X in microsomes was developed and validated according to European Medicines Agency guidelines. CAS X was demonstrated to be a substrate for carboxylesterases via hydrolysis reaction, with a Michaelis-Menten kinetic profile. The enzyme kinetic parameters were determined, and the intrinsic clearance was 1.7-fold higher in humans than in rats. The hepatic clearance was estimated by in vitro-in vivo extrapolation, resulting in more than 90% of the hepatic blood flow for both species. A qualitative study was also carried out for the metabolite identification by mass spectrometry and indicated the formation of the inactive metabolite CAS X dialdehyde. These findings demonstrate that CAS X is susceptible to first-pass metabolism and is a substrate for specific carboxylesterases expressed in liver, which may contribute to a reduction in antitumor activity when administered by the oral route.

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Section: Introductionmentioning

confidence: 99%

Characterization of Casearin X Metabolism by Rat and Human Liver Microsomes

et al. 2018

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“…Several in vitro methods are now in use and commercially available to investigate the metabolic fate of a chemical entity (Brandon et al 2003;Moreira et al 2017). Cellular preparations, such as liver microsomes and cultured hepatocytes, may hinder the detection of reactive metabolites, preventing them Electronic supplementary material The online version of this article (https://doi.org/10.1007/s43450-020-00084-8) contains supplementary material, which is available to authorized users.…”

Section: Introductionmentioning

confidence: 99%

Biomimetic Metabolism of Kaurenoic Acid Validated by Microsomal Reactions

Fernandes¹,

Oliveira²,

Barros³

et al. 2020

Rev. Bras. Farmacogn.

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The cytochrome P450 is a remarkable family of enzymes with widespread occurrence in animals and plants. In humans, they catalyze phase I metabolism reactions, such as oxidations and hydroxylations, playing a significant role in xenobiotics detoxification. Although mechanistic details of the catalytic cycle of cytochrome P450 are known, and human isoforms have their crystal structure solved, mimicking cytochrome P450 reactions using in vitro systems is still challenging. For this reason, biological assays using isolated enzymes or microsomal fractions are typically required to study the oxidative metabolism of new drugs before approval by regulatory agencies. Here we demonstrate that a series of metalloporphyrin catalysts generate the same oxidative product in vitro as a liver microsomal assay. The bioactive diterpene kaurenoic acid was chosen as a substrate model because it is a component of widely used phytomedicines in Brazil, such as guaco syrups and copaiba waxes. We evaluated the influence of reaction parameters such as catalyst type, oxygen donor type, solvent, and catalyst concentration on substrate consumption. Three products had their structures proposed by mass spectrometry, and among them, a dihydroxylated oxidation product was detected as a major metabolite after liver microsomal biotransformation and using porphyrin catalysts. We envision that the use of metalloporphyrins could be a useful strategy for biomimetic oxidation of kaurenic terpenes.

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“…Este mecanismo tem papel importante na cinética de compostos químicos absorvidos pelo organismo, seja para sua excreção ou para sua ativação (MOHUTSKY et al, 2008). O metabolismo compreende processos catalisados por enzimas, podendo ser dividido em duas fases, responsáveis por diferentes reações: reações de fase Ioxidação, redução e hidrólisee reações de fase IIconjugação de grupos polares diretamente ao composto absorvido ou a um produto de reações de fase I. Estudos de metabolismo in vitro podem fornecer dados sobre as vias metabólicas de fase I e fase II, os sítios moleculares mais vulneráveis à metabolização, os metabólitos que poderão ser formados in vivo, a toxicidade dos metabólitos formados e os efeitos da inibição e/ou indução enzimática no metabolismo (COLEMAN 2 , 2010apud FERNANDES, 2013NASSAR;TALAAT, 2004;MOREIRA et al, 2017). modelo é o mais empregado para avaliação de reações de oxidação na fase I e de conjugações na fase II (enzimas do CYP e do sistema UGTuridina-difosfatoglucoronil transferase -, respectivamente).…”

Section: Estudos De Metabolismo In Vitrounclassified

Estudo do metabolismo in vitro da caramboxina

Pereira¹

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In vitro metabolism studies of natural products are crucial in the early development of medicines

Cited by 6 publications

References 140 publications

Characterization of Casearin X Metabolism by Rat and Human Liver Microsomes

Characterization of Casearin X Metabolism by Rat and Human Liver Microsomes

Biomimetic Metabolism of Kaurenoic Acid Validated by Microsomal Reactions

Estudo do metabolismo in vitro da caramboxina

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