In Vitro Metabolism of Tresperimus by Human Vascular Semicarbazide-Sensitive Amine Oxidase

Claud, Philippe; Artur, Yves; Laine, Romuald

doi:10.1124/dmd.30.6.747

Cited by 4 publications

(11 citation statements)

References 30 publications

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“…Plasma stability assessments demonstrated interspecies differences that were consistent with the highest reported soluble plasma deamination activity being observed in pigs and dogs (Schwelberger, 2007). Prior studies have used human umbilical artery microsomes to demonstrate the involvement of VAP-1 in the metabolism of tresperimus (Claud et al, 2002). Since then, potent, selective VAP-1 inhibitors have been developed (Salter-Cid et al, 2005), and recombinant VAP-1 has become commercially available.…”

Section: Introductionmentioning

confidence: 52%

“…Although BA is not a physiologic substrate for VAP-1, there are numerous assays based on the oxidative deamination of BA to form BAL as a marker substrate for quantifying various amine oxidase activities, including radiometric detection of extracted 14 C-BAL (Lizcano et al, 1998), HPLC with fluorometric detection of derivatized BAL (van Dijk et al, 1995), UV monitoring of BAL (Heuts et al, 2011), and measurement of the H 2 O 2 produced using Amplex red (Holt et al, 1997). A common method used for the measurement of aldehydes is an acid-catalyzed derivatization with DNPH followed by HPLC with UV or fluorometric detection (Claud et al, 2002). A direct-injection LC-MS/MS method of the DNPH-derivatized BAL (BALDNP) was developed to measure the amount of BAL formed per minute per milligram of protein in in vitro incubations.…”

Section: In Vitro Vap-1 Oxidative Deamination Activitymentioning

confidence: 99%

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Oxidative Deamination of Emixustat by Human Vascular Adhesion Protein-1/Semicarbazide-Sensitive Amine Oxidase

2019

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Emixustat potently inhibits the visual cycle isomerase retinal pigment epithelium protein 65 (RPE65) to reduce the accumulation of toxic bisretinoid by-products that lead to various retinopathies. Orally administered emixustat is cleared rapidly from the plasma, with little excreted unchanged. The hydroxypropylamine moiety that is critical in emixustat's inhibition of RPE65 is oxidatively deaminated to three major carboxylic acid metabolites that appear rapidly in plasma. These metabolites greatly exceed the plasma concentrations of emixustat and demonstrate formation-rate-limited metabolite kinetics. This study investigated in vitro deamination of emixustat in human vascular membrane fractions, plasma, and recombinant human vascular adhesion protein-1 (VAP-1), demonstrating single-enzyme kinetics for the formation of a stable aldehyde intermediate in all in vitro systems. The in vitro systems used herein established sequential formation of the major metabolites with addition of assay components for aldehyde dehydrogenase and cytochrome P450. Reaction phenotyping experiments using selective chemical inhibitors and recombinant enzymes of monoamine oxidase, VAP-1, and lysyl oxidase showed that only VAP-1 deaminated emixustat. In individually derived human vascular membranes from umbilical cord and aorta, rates of emixustat deamination were highly correlated to VAP-1 marker substrate activity (benzylamine) and VAP-1 levels measured by enzyme-linked immunosorbent assay. In donormatched plasma samples, soluble VAP-1 activity and levels were lower than in aorta membranes. A variety of potential comedications did not strongly inhibit emixustat deamination in vitro. This work was supported by Acucela Inc., and has not been previously presented. https://doi.org/10.1124/dmd.118.085811. s This article has supplemental material available at dmd.aspetjournals.org. human MAO; rhVAP-1, recombinant human VAP-1; RPE65, retinal pigment epithelium protein 65; SSAO, semicarbazide-sensitive amine oxidase; VAP-1, vascular adhesion protein-1. 504

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Section: Introductionmentioning

confidence: 52%

Section: In Vitro Vap-1 Oxidative Deamination Activitymentioning

confidence: 99%

Oxidative Deamination of Emixustat by Human Vascular Adhesion Protein-1/Semicarbazide-Sensitive Amine Oxidase

2019

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“…Other structural clues include carboxylic acid16 or phenol17 groups, which can be problematic due to their potential metabolic conversion into thermally unstable glucuronide conjugates. Many other potentially problematic structural motifs have been reported 6–11, 13, 18–25, 29, 31–39…”

Section: Resultsmentioning

confidence: 99%

“…Metabolites with similar molecular masses to the parent compound have the potential to interfere with LC/MS/MS quantification of the parent via isotopic contributions of the metabolite. For example, the oxidation of a primary amine moiety to an aldehyde is accompanied by the loss of 1 Da 22. The [M + 1] isotopic contribution of the aldehyde metabolite is isobaric with the parent compound, which can potentially lead to inaccurate quantification of the latter.…”

mentioning

confidence: 99%

Identifying and overcoming bioanalytical challenges associated with chlorine-containing dehydrogenation metabolites

Furlong

Wujcik

et al. 2010

Rapid Commun. Mass Spectrom.

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Liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) is a widely utilized analytical tool for quantifying small molecules in complex biological matrices. In certain situations the mass-selection capabilities of the tandem mass spectrometer may be insufficient to discriminate between the analyte of interest and its metabolites, particularly those metabolites that are isobaric with the analyte. One scenario by which isobaric interference may occur is the metabolism of a chlorine- or bromine-containing small molecule to a metabolite with the concomitant loss of 2 Da. This report describes the detection and characterization of two distinct dehydrogenation [M-2] metabolites during LC/MS/MS quantification of a chlorinated small molecule in rat plasma samples derived from a toxicokinetic study. The potential isotope-related impact of these metabolites on quantification of the parent compound was assessed. Several alternate precursor ion and product ion combinations were evaluated and shown to minimize the quantitative impact of the interfering metabolites without having to rely on their stringent chromatographic resolution from the parent compound. These results indicate that when quantifying chlorine- or bromine-containing small molecules from in vivo samples or in vitro metabolic incubations: (1) efforts to detect potential dehydrogenation metabolites should be undertaken and (2) if such metabolites are detected, the judicious choice of alternate multiple-reaction monitoring (MRM) transitions can limit their impact on quantification of the parent molecule without the need for robust chromatographic resolution.

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“…These metabolites can potentially interfere with LC/MS/MS quantification of the analyte via contribution to the analyte MS/MS response as a result of the isobaric nature of the [M + 1] isotope of the metabolite. For example, the oxidation of a primary amine moiety to an aldehyde is accompanied by the loss of 1 Da 5. The [M + 1] isotope of the aldehyde metabolite is isobaric with the parent compound, which could potentially lead to inaccurate quantification of the latter.…”

mentioning

confidence: 99%

Use of high‐resolution mass spectrometry to investigate a metabolite interference during liquid chromatography/tandem mass spectrometric quantification of a small molecule in toxicokinetic study samples

Furlong

Bessire

Song

et al. 2010

Rapid Comm Mass Spectrometry

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During routine liquid chromatography/tandem mass spectrometric (LC/MS/MS) bioanalysis of a small molecule analyte in rat serum samples from a toxicokinetic study, an unexpected interfering peak was observed in the extracted ion chromatogram of the internal standard. No interfering peaks were observed in the extracted ion chromatogram of the analyte. The dose-dependent peak area response and peak area response versus time profiles of the interfering peak suggested that it might have been related to a metabolite of the dosed compound. Further investigation using high-resolution mass spectrometry led to unequivocal identification of the interfering peak as an N-desmethyl metabolite of the parent analyte. High-resolution mass spectrometry (HRMS) was also used to demonstrate that the interfering response of the metabolite in the multiple reaction monitoring (MRM) channel of the internal standard was due to an isobaric relationship between the (13)C-isotope of the metabolite and the internal standard (i.e., common precursor ion mass), coupled with a metabolite product ion with identical mass to the product ion used in the MRM transition of the internal standard. These results emphasize (1) the need to carefully evaluate internal standard candidates with regard to potential interferences from metabolites during LC/MS/MS method development, validation and bioanalysis of small molecule analytes in biological matrices; (2) the value of HRMS as a tool to investigate unexpected interferences encountered during LC/MS/MS analysis of small molecules in biological matrices; and (3) the potential for interference regardless of choice of IS and therefore the importance of conducting assay robustness on incurred in vitro or in vivo study samples.

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In Vitro Metabolism of Tresperimus by Human Vascular Semicarbazide-Sensitive Amine Oxidase

Cited by 4 publications

References 30 publications

Oxidative Deamination of Emixustat by Human Vascular Adhesion Protein-1/Semicarbazide-Sensitive Amine Oxidase

Oxidative Deamination of Emixustat by Human Vascular Adhesion Protein-1/Semicarbazide-Sensitive Amine Oxidase

Identifying and overcoming bioanalytical challenges associated with chlorine-containing dehydrogenation metabolites

Use of high‐resolution mass spectrometry to investigate a metabolite interference during liquid chromatography/tandem mass spectrometric quantification of a small molecule in toxicokinetic study samples

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