Evaluation of the therapeutic effects of AGA drugs by measuring finasteride, dutasteride, and dihydrotestosterone in hair.

BACKGROUND: Androgenetic alopecia (AGA) is treated by 5α-reductase inhibitors (5ARI) such as finasteride and dutasteride, which are widely used as therapeutic agents. However, their pharmacokinetics in target organs (scalp and hair follicles) have not yet been investigated. PURPOSE: To confirm the effective action of finasteride and dutasteride in the hair follicle tissues, we developed a method to measure these concentrations in hair. RESULTS: Compared to the non-detection (N.D.) group, the dihydrotestosterone (DHT) concentrations decreased significantly in both the finasteride and dutasteride groups. The dutasteride group showed significantly lower DHT concentrations among all groups. CONCLUSIONS: Measurement of finasteride, dutasteride, and DHT concentrations in hair would aid in evaluating the drug pharmacokinetics and its therapeutic effects on AGA patients.

Serum diiodotyrosine - a biomarker to differentiate destructive thyroiditis from Graves' disease.

OBJECTIVE: Conventional diagnostic methods are limited in their ability to differentiate destructive thyroiditis from Graves' disease. We hypothesised that serum diiodotyrosine (DIT) and monoiodotyrosine (MIT) levels could be biomarkers for differentiating destructive thyroiditis from Graves' disease. DESIGN: Patients with destructive thyroiditis (n = 13) and Graves' disease (n = 22) were enrolled in this cross-sectional study. METHODS: We assayed the serum DIT and MIT levels using liquid chromatography-tandem mass spectrometry. A receiver operating characteristic (ROC) curve analysis was used to determine the sensitivity and specificity of the serum DIT and MIT levels as biomarkers for differentiating destructive thyroiditis from Graves' disease. RESULTS: The serum DIT and MIT levels were significantly higher in patients with destructive thyroiditis than in those with Graves' disease. The ROC curve analysis showed that the serum DIT levels (≥359.9 pg/mL) differentiated destructive thyroiditis from Graves' disease, significantly, with 100.0% sensitivity and 95.5% specificity (P < 0.001). The diagnostic accuracy of the serum MIT levels (≥119.4 pg/mL) was not as high as that of the serum DIT levels (sensitivity, 84.6%; specificity, 77.3%; P = 0.001). CONCLUSIONS: The serum DIT levels may serve as a novel diagnostic biomarker for differentiating destructive thyroiditis from Graves' disease.

Analysis of hair steroid hormone concentrations at different parts of the head by liquid chromatography-tandem mass spectrometry.

BACKGROUND: Steroid hormones are known to be associated with diseases like androgenetic alopecia (AGA) resulting in hair loss. The lack of a detailed analysis of the local concentration of steroids in different parts of the head underlies the rationale and purpose of this study. METHODS: To evaluate the concentration distributions of steroid hormones in hair in different parts of the head, hair samples of 8 healthy men from 9 point-areas covering the frontal, parietal, and occipital regions were collected. Eight steroid hormones were measured by using the LC-MS/MS and region-wise comparison for different hormones was done using the mean z-score and Tukey's HSD. RESULTS: Five of the 8 hormones had a high concentration in the parietal region, with dihydrotestosterone (DHT) showing a peak in the central parietal region (z = 1.59) suggesting a correlation with AGA's clinical presentation. Whereas, no significant differences were observed for testosterone and cortisol between the parietal and occipital regions. Higher DHT levels at the parietal region were also verified with a small group of AGA patients. CONCLUSIONS: This research expands upon the role of steroid hormones in hair follicle tissue elucidating their relationship with disease, thus contributing to disease management.

Measurement of steroid hormones by liquid chromatography-tandem mass spectrometry with small amounts of hair.

Steroid hormone levels in hair reflect the integrated values (average values) of hormone secretion over the past few months. We have used a method to evaluate diseases and chronic stress, discrimination of banned drug use, and so on. In contrast, the hair analysis methods reported so far required at least 10 mg (about 50 to 100 hair strands) of hair to analyze multiple steroid hormones from the same sample. Here, we developed a new method for measuring steroid hormones in hair by liquid chromatography-tandem mass spectrometry, which identifies multiple steroid hormones from 5 to 10 (about 1 mg) hair strands. Ten steroid hormones (cortisol, cortisone, testosterone, dihydrotestosterone, dehydroepiandrosterone, androstenedione, progesterone, pregnenolone, androstenediol and estradiol) covering from sex hormones to stress hormones were derivatized and measured by four different measuring systems. The method showed good linearity for all steroids with correlation coefficients of 0.999 or more. The accuracy and precision of intra- and inter-assay ranged from 96.0 to 106.4% and 4.8 to 8.1% for intra-assay, and from 96.9 to 104.9% and 6.9 and 10.6% for inter-assay, respectively. A mixed solution containing 0.1 M trifluoroacetic acid and 50% acetonitrile was used to extract hair and to enhance the cortisol extraction efficiency approximately twice compared to the previously reported extraction with methanol. This method has the potential to clarify the relationship between steroid hormone levels and diseases that show alopecia such as chronic stress and androgenetic alopecia.

Analysis of the disposition of a novel p38 MAPK inhibitor, AKP-001, and its metabolites in rats with a simple physiologically based pharmacokinetic model.

5-[(2-Chloro-6-fluorophenyl)acetylamino]-3-(4-fluorophenyl)-4-(4-pyrimidinyl)isoxazole (AKP-001) is a potent p38 mitogen-activated protein kinase inhibitor that is being developed to specifically target the intestines for the treatment of inflammatory bowel disease. According to the ante-drug concept, AKP-001 was designed to be metabolized to inactive forms via the first-pass metabolism to avoid undesirable systemic exposure. The purpose of this study is to investigate the pharmacokinetic characteristics of AKP-001 and its metabolites (M1 and M2) in rats, utilizing a simple physiologically based pharmacokinetic (PBPK) model. In vitro metabolic activity of AKP-001 in the S9 fraction of rat liver was examined, and plasma concentration-time profiles were developed following intravenous and/or oral administration of AKP-001 and its metabolites. AKP-001 was primarily metabolized to M1; however, M2 was not detected in liver S9 fractions. In accordance with this observation in vitro, M2 was detected in plasma after oral dosing of AKP-001 with a lag time of 1.5 hours, but not after intravenous dosing. To analyze pharmacokinetics in rats in vivo, a simple PBPK model was developed by simultaneous fitting of the plasma concentrations after treatment with AKP-001 and its metabolites. The observed plasma concentration-time profiles of AKP-001 and metabolites were described by the model adequately. Intestinal and systemic exposures of AKP-001 were simulated using the model to assess the relationship between pharmacokinetics and efficacy/safety. Model analysis suggested that oral bioavailability of intestine-targeting ante-drugs should be low to avoid systemic side effects. The pharmacokinetic properties of AKP-001 meet this criterion owing to extensive first-pass metabolism.