An in vitro approach for comparative interspecies metabolism of agrochemicals

Whalley, Paul; Bartels, Michael; Bentley, Karin S.; Corvaro, Marco; Funk, Dorothee; Himmelstein, Matthew W.; Neumann, Birgit; Strupp, Christian; Zhang, Fagen; Mehta, Jyotigna

doi:10.1016/j.yrtph.2017.03.020

Cited by 16 publications

(6 citation statements)

References 21 publications

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“…Being commercially available, liver S9 fractions are easily accessible and reliable. One of the advantages of using the S9 mix is that co-factors are already included and therefore is ready to use upon purchase [59]. Although this is a tremendous benefit when studying drug metabolism and DDIs in vitro, the S9 mix shows direct toxicity to the cells at the necessary working concentration [60].…”

Section: Liver S9 Fractionsmentioning

confidence: 99%

Application of In Vitro Metabolism Activation in High-Throughput Screening

Ooka

Lynch

Xia

2020

IJMS

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In vitro methods which incorporate metabolic capability into the assays allow us to assess the activity of metabolites from their parent compounds. These methods can be applied into high-throughput screening (HTS) platforms, thereby increasing the speed to identify compounds that become active via the metabolism process. HTS was originally used in the pharmaceutical industry and now is also used in academic settings to evaluate biological activity and/or toxicity of chemicals. Although most chemicals are metabolized in our body, many HTS assays lack the capability to determine compound activity via metabolism. To overcome this problem, several in vitro metabolic methods have been applied to an HTS format. In this review, we describe in vitro metabolism methods and their application in HTS assays, as well as discuss the future perspectives of HTS with metabolic activity. Each in vitro metabolism method has advantages and disadvantages. For instance, the S9 mix has a full set of liver metabolic enzymes, but it displays high cytotoxicity in cell-based assays. In vitro metabolism requires liver fractions or the use of other metabolically capable systems, including primary hepatocytes or recombinant enzymes. Several newly developed in vitro metabolic methods, including HepaRG cells, three-dimensional (3D) cell models, and organ-on-a-chip technology, will also be discussed. These newly developed in vitro metabolism approaches offer significant progress in dissecting biological processes, developing drugs, and making toxicology studies quicker and more efficient.

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Section: Liver S9 Fractionsmentioning

confidence: 99%

Application of In Vitro Metabolism Activation in High-Throughput Screening

Ooka

Lynch

Xia

2020

IJMS

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“…The major metabolites identified in the in vivo samples were the acetyl-conjugates, the phase 2 metabolites of the benzyl-amine moiety of the vipadenant. The reason for the difference of in vitro/vivo metabolites would be likely due to the limitation of metabolic enzymes or cofactors existing in the in vitro liver microsome compared to the whole in vivo system [28,29,30,31,32].…”

Section: Discussionmentioning

confidence: 99%

Analysis of Vipadenant and Its In Vitro and In Vivo Metabolites via Liquid Chromatography-Quadrupole-Time-of-Flight Mass Spectrometry

et al. 2018

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A simple and sensitive liquid chromatography–quadrupole-time-of-flight–mass spectrometric (LC-QTOF-MS) assay has been developed for the evaluation of drug metabolism and pharmacokinetics (PK) properties of vipadenant in rat, a selective A2a receptor antagonist as one of the novel immune checkpoint inhibitors. A simple protein precipitation method using acetonitrile was used for the sample preparation and the pre-treated samples were separated by a reverse-phase C18 column. The calibration curve was evaluated in the range of 3.02 ~ 2200 ng/mL and the quadratic regression (weighted 1/concentration) was used for the best fit of the curve with a correlation coefficient ≥0.997. The in vivo PK studies in rats showed that vipadenant bioavailability was 30.4 ± 8.9% with a low to moderate drug clearance. In addition, in vitro/in vivo metabolite profiles in rat were also explored. Five different metabolites were observed in our experimental conditions and the major metabolites were different between in vitro and in vivo conditions. As far as we know, there has been no report on the development of quantitative methods for its PK samples nor the identification of its metabolites since vipadenant was developed. Therefore, this paper would be very useful to better understand the pharmacokinetic and drug metabolism properties of vipadenant in rat as well as other species.

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“…For the above‐mentioned reasons, the design of 3 × 3 × 3 (concentrations, time points, technical replicates of pooled test systems) would allow a reasonable setting to identify UHM and DHM, which may be less likely with simpler designs such as 1 concentration and incubation time (Whalley et al, 2017 ).…”

Section: Overview Of the Structure And Key Points Of The Scientific Opinionmentioning

confidence: 99%

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

Hernández‐Jerez¹,

Adriaanse²,

Aldrich³

et al. 2021

EFS2

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EFSA asked the Panel on Plant Protection Products and their residues to deliver a Scientific Opinion on testing and interpretation of comparative in vitro metabolism studies for both new active substances and existing ones. The main aim of comparative in vitro metabolism studies of pesticide active substances is to evaluate whether all significant metabolites formed in the human in vitro test system, as a surrogate of the in vivo situation, are also present at comparable level in animal species tested in toxicological studies and, therefore, if their potential toxicity has been appropriately covered by animal studies. The studies may also help to decide which animal model, with regard to a particular compound, is the most relevant for humans. In the experimental strategy, primary hepatocytes in suspension or culture are recommended since hepatocytes are considered the most representative in vitro system for prediction of in vivo metabolites. The experimental design of 3 9 3 9 3 (concentrations, time points, technical replicates, on pooled hepatocytes) will maximise the chance to identify unique (UHM) and disproportionate (DHM) human metabolites. When DHM and UHM are being assessed, test item-related radioactivity recovery and metabolite profile are the most important parameters. Subsequently, structural characterisation of the assigned metabolites is performed with appropriate analytical techniques. In toxicological assessment of metabolites, the uncertainty factor approach is the first alternative to testing option, followed by new approach methodologies (QSAR, read-across, in vitro methods), and only if these fail, in vivo animal toxicity studies may be performed. Knowledge of in vitro metabolites in human and animal hepatocytes would enable toxicological evaluation of all metabolites of concern, and, furthermore, add useful pieces of information for detection and evaluation of metabolites in different matrices (crops, livestock, environment), improve biomonitoring efforts via better toxicokinetic understanding, and ultimately, develop regulatory schemes employing physiologically based or physiology-mimicking in silico and/or in vitro test systems to anticipate the exposure of humans to potentially hazardous substances in plant protection products.

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An in vitro approach for comparative interspecies metabolism of agrochemicals

Cited by 16 publications

References 21 publications

Application of In Vitro Metabolism Activation in High-Throughput Screening

Application of In Vitro Metabolism Activation in High-Throughput Screening

Analysis of Vipadenant and Its In Vitro and In Vivo Metabolites via Liquid Chromatography-Quadrupole-Time-of-Flight Mass Spectrometry

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

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