GC-MS–Based Quantitative Signatures of Cytochrome P450–Mediated Steroid Oxidation Induced by Rifampicin

Moon, Ju Yeon; Kang, Se Mi; Lee, Jeongae; Cho, Joo Youn; Moon, Myeong Hee; Jang, In Jin; Chung, Bong Chul; Choi, Man Ho

doi:10.1097/ftd.0b013e318286ee02

Cited by 15 publications

(5 citation statements)

References 41 publications

(73 reference statements)

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“…Eluent was dried and resuspended in 100 l of derivitizing reagent prepared as a 500:4:2 (v/w/w) mixture of N-methyl-N-(trimethylsilyl)-trifluoroacetamide/ammonium iodide/dithiothreitol and heated to 60°C for 40 min. Derivatized samples were injected onto GC/MS (3-l injection for lyase samples and 5-l injections for hydroxylase samples) and analyzed in selected ion monitoring mode using instrument parameters as described (44) except that the column was Agilent DB-5MS, 15 ϫ 0.25-mm inner diameter with 0.25-micron film. Silylated derivatives of products 17␣-hydroxypregnenolone (retention time ϳ16.5 min; m/z ϭ 548) and dehydroepiandrosterone (retention time, ϳ7.9 min; m/z ϭ 432) were quantified based on their molecular ion with respect to that of silylated estriol internal standard (retention time ϳ15.7 min; m/z ϭ 504).…”

Section: Methodsmentioning

confidence: 99%

Structures of Human Steroidogenic Cytochrome P450 17A1 with Substrates

Petrunak

DeVore

Porubsky

et al. 2014

Journal of Biological Chemistry

109

167

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Background: Structural information for substrate binding to human steroidogenic cytochrome P450 17A1 (CYP17A1) is unavailable. Results: Within a common overall orientation, different steroids adopt subtly different positions. Conclusion: Steric and hydrogen-bonding modulation of lateral/vertical orientation controls CYP17A1-mediated steroid oxidation. Significance: Understanding the CYP17A1 mechanism provides opportunities for better targeted drug design.

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Section: Methodsmentioning

confidence: 99%

Structures of Human Steroidogenic Cytochrome P450 17A1 with Substrates

Petrunak

DeVore

Porubsky

et al. 2014

Journal of Biological Chemistry

109

167

View full text Add to dashboard Cite

show abstract

“…Free steroids without deconjugation steps were also evaluated in this study. Analytical sensitivities were improved for most androgens and estrogens (4 to 10 times), while those of progestogens and corticoids were similar or 2‐4 times more sensitive than results obtained from our previous method 15 . However, several progestogens and corticoids including Preg and F and their hydroxylated metabolites show better analytical sensitivities in a method coupled with supported liquid extraction purification 17 …”

Section: Discussionmentioning

confidence: 79%

“…Analytical sensitivities were improved for most androgens and estrogens (4 to 10 times), while those of progestogens and corticoids were similar or 2-4 times more sensitive than results obtained from our previous method. 15 However, several progestogens and corticoids including Preg and F and their hydroxylated metabolites show better analytical sensitivities in a method coupled with supported liquid extraction purification. 17 Although a lineage association between fetal and adult Leydig cells has not been fully identified, 20 fetal Leydig cells are known to persist up to 30% of total Leydig cells in adult testes.…”

Section: Discussionmentioning

confidence: 99%

“…In this study, gas chromatography-mass spectrometry (GC-MS)based quantitative profiling 14,15 was further developed and applied to measure localized concentrations of 23 androgens, 7 estrogens, 14 progestogens, and 13 corticoids in both fetal and adult mouse testes. These quantitative results may reveal testicular metabolic signatures including steroidogenic and oxidation pathways along with male development.…”

Section: Introductionmentioning

confidence: 99%

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GC‐MS‐based metabolic signatures reveal comparative steroidogenic pathways between fetal and adult mouse testes

et al. 2020

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Background Previous studies on gonadal steroidogenesis have not compared metabolic pathways between fetal and adult mouse testes to date. Objectives To evaluate comparative metabolic signatures of testicular steroids between fetus and adult mice using gas chromatography‐mass spectrometry (GC‐MS)‐based steroid profiling. Materials and methods GC‐MS with molecular‐specific scan modes was optimized for selective and sensitive detection of 23 androgens, 7 estrogens, 14 progestogens, and 13 corticoids from mouse testes with a quantification limit of 0.1‐5.0 ng/mL and reproducibility (coefficient of variation: 0.3%‐19.9%). Based on 26 steroids quantitatively detected in testes, comparative steroid signatures were analyzed for mouse testes of 8 fetuses on embryonic day 16.5 and 8 adults on postnatal days 56‐60. Results In contrast to large amounts of steroids in adult testes (P < .0002), all testicular levels per weight unit of protein were significantly increased in fetal testes (P < .002, except 6β‐hydroxytestosterone of P = .065). Both 11β‐hydroxyandrostenedione and 7α‐hydroxytestosterone were only measurable in fetal testes, and metabolic ratios of testosterone to androstenediol and androstenedione were also increased in fetal testes (P < .05 for both). Discussion and conclusion Testicular steroid signatures showed that both steroidogenic Δ4 and Δ5 pathways in the production of testosterone were activated more during prenatal development. Both 7α‐ and 11β‐hydroxylations were predominant, while hydroxylations at C‐6, C‐15, and C‐16 of testosterone and androstenedione were decreased in the fetus. The present GC‐MS‐based steroid profiling may facilitate understanding of the development of testicular steroidogenesis.

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“…The concentration of five distinct endogenous steroids (four in urine and one in plasma samples) was quantified by gas chromatography and triple quadrupole mass spectrometry. The urine concentration of cortisol, 6β-OH-cortisol, cortisone, and 6β-OH-cortisone19 and the plasma concentration of 4β-OH-cholesterol20 were quantified using previously described protocols, with minor modifications.…”

Section: Methodsmentioning

confidence: 99%

Assessment of Hepatic Cytochrome P450 3A Activity Using Metabolic Markers in Patients with Renal Impairment

et al. 2018

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BackgroundThe renal function of individuals is one of the reasons for the variations in therapeutic response to various drugs. Patients with renal impairment are often exposed to drug toxicity, even with drugs that are usually eliminated by hepatic metabolism. Previous study has reported an increased plasma concentration of indoxyl sulfate and decreased plasma concentration of 4β-hydroxy (OH)-cholesterol in stable kidney transplant recipients, implicating indoxyl sulfate as a cytochrome P450 (CYP) inhibiting factor. In this study, we aimed to evaluate the impact of renal impairment severity-dependent accumulation of indoxyl sulfate on hepatic CYP3A activity using metabolic markers.MethodsSixty-six subjects were enrolled in this study; based on estimated glomerular filtration rate (eGFR), they were classified as having mild, moderate, or severe renal impairment. The plasma concentration of indoxyl sulfate was quantified using liquid chromatography-mass spectrometry (LC-MS). Urinary and plasma markers (6β-OH-cortisol/cortisol, 6β-OH-cortisone/cortisone, 4β-OH-cholesterol) for hepatic CYP3A activity were quantified using gas chromatography-mass spectrometry (GC-MS). The total plasma concentration of cholesterol was measured using the enzymatic colorimetric assay to calculate the 4β-OH-cholesterol/cholesterol ratio. The correlation between variables was assessed using Pearson's correlation test.ResultsThere was a significant negative correlation between MDRD eGFR and indoxyl sulfate levels. The levels of urinary 6β-OH-cortisol/cortisol and 6β-OH-cortisone/cortisone as well as plasma 4β-OH-cholesterol and 4β-OH-cholesterol/cholesterol were not correlated with MDRD eGFR and the plasma concentration of indoxyl sulfate.ConclusionHepatic CYP3A activity may not be affected by renal impairment-induced accumulation of plasma indoxyl sulfate.

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GC-MS–Based Quantitative Signatures of Cytochrome P450–Mediated Steroid Oxidation Induced by Rifampicin

Abstract: This CYP-mediated steroid signature profile allows simultaneous assessment of CYP1A, CYP1B, CYP2C, CYP3A, CYP11B, CYP17A, CYP19A, and CYP21A in urine samples. This method could therefore be a useful tool for assessing drug efficacy.

Cited by 15 publications

References 41 publications

Structures of Human Steroidogenic Cytochrome P450 17A1 with Substrates

Structures of Human Steroidogenic Cytochrome P450 17A1 with Substrates

GC‐MS‐based metabolic signatures reveal comparative steroidogenic pathways between fetal and adult mouse testes

Assessment of Hepatic Cytochrome P450 3A Activity Using Metabolic Markers in Patients with Renal Impairment

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