Excretion Of 6β-hydroxycortisol Research Articles

Different Types of Urinary Steroid Profiling Obtained by High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectrometry in Patients with Adrenocortical Carcinoma.

Urinary steroid profiling (USP) was studied using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods in 108 patients with adrenocortical adenoma (ACA) and in 31 patients with adrenocortical carcinoma (ACC). Thirteen ACC and Cushing's syndrome (ACC-CS) patients had two types of USP as well as 18 ACC patients without hypercortisolism. These four types differed by androgen and glucocorticoid secretion of the adrenal cortex. Fifteen main ACC features were observed by GC-MS. Urinary excretion of dehydroepiandrosterone (DHEA) was increased in 67.7% of ACC patients and tetrahydro-11-deoxycortisol (THS) in 74.2%. By combination of the following parameters: THS >900μg/24h and/or DHEA >1500μg/24h with ratios of 3α,16,20-pregnentriol/3β,16,20-pregnentriol (3α,16,20dP3/3β,16,20dP3) less than 6.0 and 3α,17,20dP3/3β,17,20dP3 less than 9.0 and the detection of "non-classical" 5-en-pregnens, not found in ACA and healthy persons, 100% sensitivity and specificity of ACC and ACA differential diagnosis were achieved. Features of 21-hydroxylase and 11β-hydroxylase deficiency were observed by GC-MS in 32.2 and 61.3% of the ACC patients, respectively. Additional features for ACC-CS diagnostic were increased urinary excretion of 6β-hydroxycortisol, 18-hydroxycorticosterone, the sum (UFF + UFE) obtained by HPLC, tetrahydrocorticosterone, and the sum (THF + THE + allo-THF) obtained by GC-MS.

Effects of Seijo‐bofu‐to, a Traditional Japanese Herbal Medicine Containing Furanocoumarin Derivatives, on the Drug‐Metabolizing Enzyme Activities in Healthy Male Volunteers

Seijo-bofu-to, a traditional medicine used to treat acne in Asian countries, contains twelve herbal components, including Angelica dahurica root, a source of furanocoumarin derivatives. In this study, we investigated potential herb-drug interactions of seijo-bofu-to in healthy male volunteers. Thirty-two young, healthy, non-smoking males were assessed for the baseline activity of cytochrome P450 (CYP) 1A2, CYP3A, CYP2D6, N-acetyltransferase 2 and xanthine oxidase according to the urinary metabolic indices of 8-hr urine samples collected after the administration of a 150-mg dose of caffeine and a 30-mg dose of dextromethorphan, and the ratio of urinary excretion of 6β-hydroxycortisol to cortisol. Thereafter, the volunteers received 3.75 g of seijo-bofu-to twice daily for 7 days and underwent the same tests on post-dose day 7. The geometric mean ratio of the CYP1A2 activity on day 7 to that observed at baseline was 0.66 (95% CI, 0.55-0.79, p = 0.001). The geometric mean phenotypic indices for CYP3A, CYP2D6, N-acetyltransferase 2 and xanthine oxidase on day 7 did not differ from the baseline values. The findings of the present study suggest that seijo-bofu-to may inhibit the activity of CYP1A2, whereas it is unlikely to participate in herb-drug interactions involving medications predominantly metabolized by CYP3A, CYP2D6, N-acetyltransferase 2 or xanthine oxidase.

Mitotane Therapy in Adrenocortical Cancer Induces CYP3A4 and Inhibits 5α-Reductase, Explaining the Need for Personalized Glucocorticoid and Androgen Replacement

Mitotane [1-(2-chlorophenyl)-1-(4-chlorophenyl)-2,2-dichloroethane] is the first-line treatment for metastatic adrenocortical carcinoma (ACC) and is also regularly used in the adjuvant setting after presumed complete removal of the primary tumor. Mitotane is considered an adrenolytic substance, but there is limited information on distinct effects on steroidogenesis. However, adrenal insufficiency and male hypogonadism are widely recognized side effects of mitotane treatment. Our objective was to define the impact of mitotane treatment on in vivo steroidogenesis in patients with ACC. At seven European specialist referral centers for adrenal tumors, we analyzed 24-h urine samples (n = 127) collected from patients with ACC before and during mitotane therapy in the adjuvant setting (n = 23) or for metastatic ACC (n = 104). Urinary steroid metabolite excretion was profiled by gas chromatography/mass spectrometry in comparison with healthy controls (n = 88). We found a sharp increase in the excretion of 6β-hydroxycortisol over cortisol (P < 0.001), indicative of a strong induction of the major drug-metabolizing enzyme cytochrome P450 3A4. The contribution of 6β-hydroxycortisol to total glucocorticoid metabolites increased from 2% (median, interquartile range 1-4%) to 56% (39-71%) during mitotane treatment. Furthermore, we documented strong inhibition of systemic 5α-reductase activity, indicated by a significant decrease in 5α-reduced steroids, including 5α-tetrahydrocortisol, 5α-tetrahydrocorticosterone, and androsterone (all P < 0.001). The degree of inhibition was similar to that in patients with inactivating 5α-reductase type 2 mutations (n = 23) and patients receiving finasteride (n = 5), but cluster analysis of steroid data revealed a pattern of inhibition distinct from these two groups. Longitudinal data showed rapid onset and long-lasting duration of the observed effects. Cytochrome P450 3A4 induction by mitotane results in rapid inactivation of more than 50% of administered hydrocortisone, explaining the need for doubling hydrocortisone replacement in mitotane-treated patients. Strong inhibition of 5α-reductase activity is in line with the clinical observation of relative inefficiency of testosterone replacement in mitotane-treated men, calling for replacement by 5α-reduced androgens.

Effect of the Fluoroquinolone Antibacterial Agent DX-619 on the Apparent Formation and Renal Clearances of 6β-Hydroxycortisol, an Endogenous Probe for CYP3A4 Inhibition, in Healthy Subjects

To examine the effect of the fluoroquinolone DX-619 on CYP3A4 and urinary excretion of 6β-hydroxycortisol, an endogenous probe of hepatic CYP3A4 activity, in healthy subjects. The effect of DX-619 on CYP3A4 was examined in human liver microsomes. The apparent formation and renal clearance of 6β-hydroxycortisol (CL(6β-OHF) and CL(renal,6β-OHF), respectively) were determined in placebo- and DX-619-treated subjects. 6β-hydroxycortisol uptake was determined in HEK293 cells expressing OAT1, OAT3, OCT2, MATE1, and MATE2-K. DX-619 was a mechanism-based inhibitor of CYP3A4, with K(I) and k(inact) of 67.9 ± 7.3μmol/l and 0.0730 ± 0.0033min(-1), respectively. Pharmacokinetic simulation suggested in vivo relevance of CYP3A4 inhibition by DX-619. CL(6β-OHF) and CL(renal,6β-OHF) were decreased 72% and 70%, respectively, on day 15 in DX-619-treated group compared with placebo (P < 0.05). 6β-hydroxycortisol was a substrate of OAT3 (K(m) = 183 ± 25μmol/l), OCT2, MATE1, and MATE2-K. Maximum unbound concentration of DX-619 (9.1 ± 0.4μmol/l) was above K(i) of DX-619 for MATE1 (4.32 ± 0.79μmol/l). DX-619 caused a moderate inhibition of hepatic CYP3A4-mediated formation and significant inhibition of MATE-mediated efflux of 6β-hydroxycortisol into urine. Caution is needed in applying CL(6β-OHF) as an index of hepatic CYP3A4 activity without evaluating CL(renal,6β-OHF).

Poor correlation between 6β‐hydroxycortisol:cortisol molar ratios and midazolam clearance as measure of hepatic CYP3A activity: a comment

I read with great interest the paper of Chen and coworkers [1], who reported on the ‘relationship between a validated biomarker, intravenous midazolam clearance and the urinary cortisol ratio under constitutive conditions and with the influence of a moderate CYP3A inhibitior’ (p.1). The authors concluded that the ‘high intra-individual variability of the urinary cortisol ratio, compared with midazolam, makes this a suboptimal CYP3A phenotyping tool’ (p.1). A shortcoming may confound their results making the interpretation of their cortisol ratio difficult, if not impossible: the authors did not pay attention to a possible influence of varying urine volume. This criticism is based on the well-documented relationship between urine volume and urinary free cortisol excretion [2–8]. Unfortunately, the results of Chen et al. [1] would not be improved if results were reworked and the volume of each 24-h urine taken into calculation, since urinary steroid excretion depends also on daytime: an increase of urine volume in the morning (zenith of adrenocortical secretion [9]) will have a more pronounced effect on steroid excretion than a similar change of urine flow in the evening (nadir of adrenocortical secretion [9]). Conclusion: urinary free cortisol and probably of 6β-hydroxycortisol excretion in 24-h urines should be determined only if the urine volume is kept as constant as possible – this demands that patients or volunteers strictly observe their fluid intake (volume, daytime). Future work will then show that the ratio of urinary 6β-hydroxycortisol/cortisol can be used an index for CYP3A phenotyping.

Combined Exposure to Deet (N,N-Diethyl-m-Toluamide) and Permethrin: Pharmacokinetics and Toxicological Effects

Permethrin and DEET are concurrently used for pests control inside homes, in public places, and in military shelters. Combined exposure to these compounds produced greater biochemical, behavioral, and metabolic alterations in animals compared to each individual compound. Concurrent application of DEET and permethrin induced urinary excretion of 3-nitrotyrosine and 8-hydroxy-2′-deoxyguanosine, markers of DNA damage and oxidative stress in rats, increased the release of rat brain mitochondrial cytochrome c, disrupted the blood–brain barrier (BBB) in rats, decreased m2 muscarinic acetylcholine receptor ligand binding density in rat brain, increased urinary excretion of 6β-hydroxycortisol, a marker CYP3A4 induction, altered sensorimotor and locomotor activities in rats, and changed in vivo and in vitro metabolism and pharmacokinetic profiles of the individual compound. These findings show that more research is needed to examine adverse effects of the combined use of DEET and permethrin on other biochemical/physiological system(s) and to predict mechanistic pathways for these effects, particularly mechanism of action at cellular and molecular levels and alterations of genes transcription.

Influence of Amiodarone on Urinary Excretion of 6β-Hydroxycortisol in Humans

Influence of Amiodarone on Urinary Excretion of 6β-Hydroxycortisol in Humans

Influence of CYP3A Metabolizer Status on the Pharmacokinetics and Pharmacodynamics of Amiodarone

The present work was designed to determine whether the individual differences in pharmacokinetics and pharmacodynamics of amiodarone and its N-desethyl metabolite are related to cytochrome CYP3A metabolizer status. 12 cardiac patients with inducible ventricular tachyarrhythmias during the baseline electrophysiologic study were enrolled in this study. Urinary 24-hour excretion of 6 beta-hydroxycortisol (6 beta-OHC and the ratio of 6 beta-hydroxycortisol to urinary free cortisol (6 beta-OHC/UFC) were measured before the first amiodarone administration. Trough plasma concentrations of amiodarone and N-desethylamiodarone (N-DEA) were measured after 79 +/- 11 days (2nd period) and after 182 +/- 25 days (3rd period). Electrophysiologic effects of amiodarone therapy were established with serial electrophysiologic studies in 9 of these patients at the baseline and after 79 +/- 11 days (during the second period). Both the 6 beta-OHC excretion and 6 beta-OHC/UFC ratio varied approximately 6-fold between the patients. We found significant inverse correlation between the 6 beta-OHC excretion and the trough plasma concentrations of amiodarone at the time of the 3rd period (rs = -0.58, p < 0.05). Similarly, there was correlation between the 24-hour urinary 6 beta-OHC excretion and trough plasma concentrations of amiodarone during the 3rd period (rs = -0.64, p < 0.025). We were unable to detect any association between CYP3A activity and amiodarone pharmacodynamics. This study points toward important information value of CYP3A metabolizer status in the context of therapeutic drug monitoring of amiodarone.

Increase in urinary excretion of 6beta-hydroxycortisol in common marmosets as a marker of hepatic CYP3A induction.

The ratio of urinary 6beta-hydroxycortisol (6beta-OHF) to free cortisol (F), i.e., the 6beta-OHF/F ratio, has been reported to be a specific marker for human CYP3A induction by in vivo studies of human subjects. In the development of drugs, it is quite beneficial to predict human CYP3A induction in preclinical safety studies using urine samples from experimental animals. We examined the 6beta-OHF/F ratio in urine of common marmosets administered with rifampicin, a potent inducer of CYP3A, to evaluate the usefulness of common marmosets for the prediction of CYP3A induction. Rifampicin was orally administered to three groups of four male common marmosets at doses of 0, 10, and 20 mg/kg per day for 4 days. Amounts of 6beta-OHF and F in urine samples were determined by means of high-performance liquid chromatography (HPLC) during the experimental period. One day after the 4th dosing, animals were killed, and P450 contents and P450-catalyzed. 7-alkoxycoumarin O-dealkylase (ACD) activities in the liver were measured. Western blot analysis of liver microsomes was also performed using anti-rat P450 (CYP1A1, 2B1/2, 3A, and 4A) antibodies. The results indicated elevations in the 6beta-OHF/F ratios that were dependent on both the dosing period and dose levels adopted. The ratios on day 4 reached 4.7- and 5.3-fold the pre-administration values in the 10 and 20 mg/kg per day groups, respectively. P450 contents and ACD activities were also elevated in all of the groups. Western blot analysis showed specific increases in the protein which cross-reacts with anti-rat CYP3A antibody in all of the groups. Furthermore, the 6beta-OHF/F ratio was well correlated with the CYP3A contents in liver (r = 0.906). These results indicated that increase in urinary excretion of 6beta-OHF is a specific marker of the induction of hepatic CYP3A in common marmosets just as in humans. Consequently, the present study suggested that human CYP3A induction elicited by chemical agents can be predicted in common marmosets by measuring the urinary excretion of 6beta-OHF.

CYP 3A proteins are expressed in human neutrophils and lymphocytes but are not induced by rifampicin

Cytochrome P-450 3A (CYP 3A) enzymes, the prominent subfamily in the cytochrome system, are expressed in various extrahepatic tissues. Until now, their expression has been demonstrated in human polymorphic neutrophils (PMNs) but not in lymphocytes using immunohistochemistry and immunoblot analysis. Moreover, their potential modulation has not been determined yet. To study such an expression in different peripheral blood cell populations, rifampicin (600 mg/day during 6 days) was given to 8 healthy subjects. PMNs and lymphocytes were isolated by centrifugation of whole white blood cell fractions using Ficoll gradients before drug administration, immediately after, and 3 days after drug withdrawal. PMN and lymphocyte smears and homogenates were subjected to immunostaining and immunoblotting, respectively, with a mouse monoclonal antibody recognizing all CYP 3A proteins. These proteins were quantified by densitometric analysis. Before and after rifampicin administration, a positive cytoplasmic staining was observed in all PMNs and in about 50% of lymphocytes. CYP 3A expression in lymphocytes was further confirmed by positive immunoblots for lymphocyte homogenates. Neither in PMNs nor in lymphocytes, induction of CYP 3A protein expression was observed after rifampicin treatment despite overall induction of CYP 3A activity assessed by the urinary excretion of 6β-hydroxycortisol. These results demonstrate that CYP 3A proteins are constitutively expressed not only in PMNs but also in lymphocytes. However, in both cell lineages CYP 3A protein expression was not induced by rifampicin.

Assessment of Doxylamine Influence on Mixed Function Oxidase Activity upon Multiple Dose Oral Administration to Normal Volunteers

The primary purpose of this study was to assess the influence of doxylamine and phenobarbital on antipyrine/metabolites pharmaco-kinetics and 6β-hydroxycortisol urinary excretion. This study was conducted in 48 healthy male human volunteers (16 per treatment group) using a parallel study design. Treatment groups consisted of 12.5mg of doxylamine succinate, placebo, or 30mg of phenobarbital administered orally every 6h for 17days. Results indicate that no statistically significant differences were observed between the doxylamine and placebo groups that are indicative of enzyme induction. For the phenobarbital group, a significant increase for antipyrine total (36 versus 45mL/h/kg) and nonrenal (35 versus 44mL/h/kg) clearances and 6β-hydroxycortisol excretion (338 versus 529 µg) and a significant decrease in the terminal exponential half-life (11 versus 9 h) of antipyrine were observed.

Urinary 6 beta-hydroxycortisol excretion in Hong Kong Chinese patients with hepatocellular carcinoma and other chronic liver diseases.

The biotransformation of xenobiotics into toxic metabolites by cytochrome P-450 has been implicated in carcinogenesis. This study investigated CYP3A4 activity, which metabolically activates procarcinogens such as aflatoxin B1, by measuring the urinary 6 beta-hydroxycortisol (6 beta OHF) to free cortisol (F) ratio in patients with hepatocellular carcinoma (HCC) and other chronic liver diseases. One hundred forty-three controls and 150 patients with different liver diseases, including chronic liver disease (due to alcoholism and/or chronic hepatitis B virus infection), cirrhosis (any cause), and resectable and nonresectable HCC, were recruited. Twenty-four hour urine samples were collected for measurement of 6 beta OHF and free cortisol by an enzyme-linked immunosorbent assay (ELISA) and a radioimmunoassay, respectively. Patients with nonresectable HCC showed a significant increase in 6 beta OHF excretion as well as their 6 beta OHF/F ratio (P < 0.05) when compared with the controls and other liver disease groups including patients with resectable HCC. The nonresectable HCC group showed a bimodal distribution in the 6 beta OHF/F ratio. Using a ratio of 9 or more in all HCC patients, the sensitivity and specificity of using the 6 beta OHF/F ratio to predict nonresectability of HCC was 48.8% and 92.6%, respectively. Our results show an increase in mean CYP3A4 enzyme activity, reflected as an increase in the 6 beta OHF/F ratio, in Hong Kong Chinese with nonresectable HCC compared with those with resectable HCC and other liver diseases. Although the role of increased CYP3A4 activity in the aetiology of HCC is not known, our specificity and sensitivity estimates suggest that a high 6 beta OHF/F ratio indicates probable inoperability. However, a normal level is a poor predictor of resectability.

Urinary excretion of 6β-hydroxycortisol in women during treatment with different oral contraceptive formulations

The measurement of the urinary excretion ratio of 6β-hydroxycortisol (6β- OHC) cortisol was used as a non-invasive method to investigate possible changes in the activity of drug-metabolizing enzymes in women receiving different oral contraceptive formulations for 1 up to 3 treatment cycles. The contraceptive preparations were either levonorgestrel, gestodene or cyproterone acetate, each in combination with ethinyl estradiol, or only the progestogens levonorgestrel or gestodene. There was either no or only a small decrease in the 6β- OHC cortisol ratio. Thus, only a minor inhibitory effect, if any, can be ascribed to the investigated contraceptive steroids in vivo. Previously observed differences between selected contraceptive steroids in vitro were not observed in the same way in vivo. This may be due either to the absence of a marked inhibitory activity in vivo or to the insufficient sensitivity of the marker 6β- OHC cortisol to detect these changes. Another possible reason may be the considerably higher drug concentrations used in the in vitro studies as compared to those present in the serum of women under oral contraceptive therapy.

Effects of Simvastatin and Pravastatin on 6β-hydroxycortisol Excretion, a Potential Marker of Cytochrome P-450 3A

The influence of two inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase, simvastatin and pravastatin, on the level of urinary 6β-hydroxycortisol/17-hydroxycorticosteroids (6β-OHC/17-OHCS) ratio was determined in two groups of normolipidemic Caucasian subjects ( n= 18 and n= 14, respectively). The 6β-OHC/17-OHCS ratio increased significantly after simvastatin administration (20 mg day -1 for 17 days) ( P=0.0125) whereas no modification was observed after pravastatin administration (20 mg day -1 during 17 days). As the level of 6β-OHC/17-OHCS ratio is a function of cytochrome P-450 3A activity, these results suggest that in Caucasian subjects, simvastatin but not pravastatin could be a weak inducer of cytochrome P-450 3A. This contrasting effect could be related to the major pharmacological differences existing between these two drugs.

Estimation of urinary 6β-hydroxycortisol by "on-line" liquid chromatography

Urinary excretion of 6β-hydroxycortisol is widely used as a simple, non-invasive index of enzyme induction in man and as a biochemical marker for chronic hyperglucocorticoid states. Beside radioimmunological assays, liquid Chromatographie methods for its estimation have been described. Because of the immense challenge to eliminate interfering urinary chromogens, these techniques are very troublesome and time-consuming. Using a previously described principle of sample clean-up, we developed a fully automated, liquid Chromatographie method for 6β-hydroxycortisol estimation in human urine.

Diurnal variation in the excretion of 6 beta-hydroxycortisol and estriol at 32 weeks gestation.

We have previously shown a marked diurnal variation in the excretion of cortisol and 6 beta-hydroxycortisol at 36-38 weeks pregnancy with no variation in estrogen excretion. Since others have shown a reciprocal relationship between plasma levels of estriol and cortisol at 30-35 weeks of pregnancy, we decided to determine whether a similar relationship was present in urinary concentrations of cortisol, 6 beta-hydroxycortisol and estriol at this gestational age. A significant variation in excretion of both cortisol and 6 beta-hydroxycortisol was found. The variation in urinary levels of estriol was less than that of the corticosteroids but followed the same pattern.

The urinary 6β-hydroxycortisol excretion in man on inducers and inhibitors of the hepatic mixed function oxidase

The urinary 6β-hydroxycortisol excretion in man on inducers and inhibitors of the hepatic mixed function oxidase

Can the urinary excretion of 6β hydroxycortisol allow for the prediction of the optimal posology of cyclosporin A in transplant recipients

Can the urinary excretion of 6β hydroxycortisol allow for the prediction of the optimal posology of cyclosporin A in transplant recipients

Age- and sex-related differences in urinary excretion of 6β-hydroxycortisol in humans

A study of normal subjects showed that the 24-h urinary 6β-hydroxycortisol level is age-dependent, with a maximum at 14–20 yr in both men and women. The ratio of 6β-hydroxycortisol cortisol also showed an age-dependent variation, being higher in children than adults. There was a significant difference between boys and girls aged 5–10 yr, but no difference in other subjects. No sex difference was demonstrable in the ratio of 6β-hydroxycortisol cortisol . Examination of circadian rhythm revealed that 6β-hydroxycortisol and cortisol correlated well in children, but showed poor correlation in adults, with a lowered morning peak of 6β-hydroxycortisol.

The effect of spironolactone on the liver microsomal enzyme system in patients.

British Journal of Clinical PharmacologyVolume 17, Issue 4 p. 485-486 Free Access The effect of spironolactone on the liver microsomal enzyme system in patients. EE Ohnhaus, EE OhnhausSearch for more papers by this authorE Gerber-Taras, E Gerber-TarasSearch for more papers by this author EE Ohnhaus, EE OhnhausSearch for more papers by this authorE Gerber-Taras, E Gerber-TarasSearch for more papers by this author First published: April 1984 https://doi.org/10.1111/j.1365-2125.1984.tb02378.xCitations: 2AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References Feller, D. R. & Gerald, M. C. (1971). Interactions of spironolactone with hepatic microsomal drug metabolising systems. Biochem. Pharmac., 20, 1991– 2000. Huffman, D. H., Shoeman, D. W., Pentikäinen, P. & Azarnoff, D. L. (1973). The effect of spironolactone on antipyrine metabolism in man. Pharmacology, 10, 338– 344. Ohnhaus, E. E. & Park, B. K. (1979). Measurement of urinary 6β-hydroxycortisol excretion as an in vivo parameter in the clinical assessment of the microsomal enzyme inducing capacity of antipyrine, phenobarbitone and rifampicin. Eur. J. clin. Pharmac., 15, 139– 145. Talcott, R. E. & Stohs, S. J. (1972). Metabolism of aniline and hexobarbital by liver homogenates of spironolactone pretreated rats. J. pharm. Sci., 61, s296– 297. Taylor, S. A., Rawlins, M. D. & Smith, S. E. (1972). Spironolactone—a weak inducer in man. J. Pharm. Pharmac., 24, 578– 579. Citing Literature Volume17, Issue4April 1984Pages 485-486 ReferencesRelatedInformation