In this study, the degradation behavior and reaction mechanisms of triazolobenzodiazepines (TBZDs)-alprazolam (ALZ), triazolam, and etizolam-in artificial gastric juice were investigated. Degradation kinetics were evaluated by monitoring time-dependent changes in TBZD standard solutions after the addition of artificial gastric juice at various temperatures using LC with photodiode array detection. Arrhenius analysis provided kinetic parameters for each compound. All TBZDs underwent rapid degradation under acidic conditions, and the reaction was reversible, yielding a single major product that was characterized as the final degradation product. Structural elucidation of the ALZ degradation product by spectroscopic and mass spectrometric analyses identified it as (2-[3-(aminomethyl)-5-methyl-4H-1,2,4-triazol-4-yl]-5-chlorophenyl)(phenyl)methanone. To gain mechanistic insight, quantum chemical calculations were performed, including geometry optimization, electrostatic potential analysis, and frontier molecular orbital analysis. These analyses indicated that protonation enhances the electrophilicity of the azomethine carbon, facilitating nucleophilic attack by water. These findings demonstrate that electronic structure analysis based on quantum chemical calculations provides valuable mechanistic insight into the hydrolysis of TBZDs under acidic conditions and may contribute to the rational assessment of chemical stability in future drug development.

To address the lack of visual detection methods for the screening of abused benzodiazepines (BZDs), this study proposes a biomimetic probe design strategy. Specifically, molecular docking simulations of the γ-aminobutyric acid type-A receptor and BZD complexes revealed multiple shared π … π interactions with the TYR-58, PHE-77, PHE-100, and TYR-160 residues, along with a hydrophobic interaction with the VAL-203. The emulation of these receptor-ligand π … π interactions guided the biomimetic design of the styrylbenzothiazole-based fluorescent probes featuring a large π-conjugation structure. The optimal probe (E)-4-(2-(5-methoxybenzo[d]thiazol-2-yl)vinyl)benzonitrile (BCN), recognizes BZDs by forming π … π interactions with its phenyl/triazole moieties and weak hydrogen bond interactions with the imine and triazole nitrogen atoms, or carbonyl oxygen. Moreover, the BCN-functionalized sensing chip enables ratiometric fluorescence detection of typical BZDs (e.g., bromazolam, triazolam, diazepam, and oxazepam) via a target-assisted rightward shift of the excited-state cis-to-trans isomerization equilibrium. And this chip could enable visual qualitative detection of diazepam in actual tablet samples and four typical BZDs in complex matrices such as urine and wastewater.

1. Traditional Japanese medicines (Kampo medicines) are increasingly used concurrently with other medications, but evidence regarding their potential for drug-drug interactions (DDIs) is insufficient. Given the important role of cytochrome P450 3 A (CYP3A) in DDIs, this study assessed the CYP3A-mediated DDI risk of three Kampo medicines (Hochuekkito, Goreisan, and Ninjin'yoeito).2. We utilised a CYP3A-humanised mouse model to comprehensively evaluate the effects of the three Kampo medicines on CYP3A activity, including an analysis of the area under the plasma concentration-time curve (AUC) and AUC ratio (AUCR).3. None of the three Kampo medicines tested significantly affected the AUC of the CYP3A substrate triazolam or the AUCRs of triazolam metabolites.4. These results provide important preclinical evidence indicating the lack of a significant net effect on CYP3A-mediated DDIs for Hochuekkito, Goreisan, and Ninjin'yoeito under the tested repeated-dosing conditions, thereby supporting their safe and proper use.

Ketoconazole (KC), a potent cytochrome P450 (CYP) 3A inhibitor, is hydrolyzed by arylacetamide deacetylase (AADAC) in the liver to N-deacetylketoconazole (DAK). Both KC and DAK are known to inhibit CYP3A in human liver microsomes, and DAK accumulates extensively in the liver. We investigate the impact of AADAC on KC-induced drug-drug interactions (DDI) in vivo. DAK inhibited Cyp3a-mediated triazolam hydroxylation in mouse liver microsomes, whose potency was lower than KC (Ki was 0.11 ± 0.03 μM vs 0.008 ± 0.002 μM). Upon KC administration (20 mg/kg, p.o.) to wild-type mice, KC was cleared from the liver within 8 h, whereas DAK remained after 24 h. These findings suggested a smaller impact on DDI in Aadac knockout mice than in wild-type mice. However, the plasma AUC of triazolam (0.5 mg/kg, p.o.) following KC preadministration was significantly higher in Aadac knockout mice than in wild-type mice, indicating that accumulated hepatic DAK in wild-type mice does not affect triazolam hydroxylation. This can be explained by preferential DAK accumulation in lipid-rich organelles, limiting its effective cytosolic concentration to inhibit Cyp3a. In conclusion, Aadac attenuated KC-related DDIs by increasing DAK, which preferentially accumulated in intracellular organelles and thus had a limited impact on cytosolic Cyp3a inhibition.

Purpose: This retrospective study assessed the presence of adverse events during moderate sedation for pediatric dental procedures performed in a dental clinic. Methods: Dental treatment completed with sedation between June 2014 to November 2024 was assessed for the presence of any adverse events (AEs). AEs were classified by the modified Tracking and Reporting Outcomes of Procedural Sedation (TROOPS) scale and an adverse event list from the Pediatric Sedation Research Consortium (PSRC). Results: There were 923 completed patient sedation records between the ages of 2 and 16 years from 9 different sedation regimens within the dental clinic included in this study. Overall, the proportion of adverse events was low (4.55%) with an increase in AE with the intranasal dexmedetomidine/orally administered midazolam (odds ratio [OR]=4.7, P<0.01) and a decrease in AE with triazolam (OR equals 0.2, P<0.01). No adverse events required medical intervention. Conclusions: Overall, adverse event proportions were low with pediatric dental moderate sedation supporting the safety of this modality of treatment. More studies are needed to assess adverse events and adverse event scales in pediatric dental sedation.

Prolonged immersion in a reheated bathtub can cause extensive thermal alteration and produce intracranial findings that mimic traumatic lesions on postmortem computed tomography (PMCT). Here, we report a bath-related fatality in a woman in her late 60s in which PMCT demonstrated a crescent-shaped epidural collection along the right parietal convexity. The collection exhibited layered components, consisting of a fat-attenuation upper layer and a dependent low-attenuation layer with a small amount of gas. PMCT also demonstrated diffuse bilateral ground-glass opacities, bilateral pleural effusions, and left pneumothorax, as well as fluid accumulation in the paranasal sinuses and airway fluid with foam. At autopsy, extensive epidermal stripping and blistering with heat rigor were observed, and red frothy fluid was present within the airway. Autopsy revealed a brick-colored epidural deposit (approximately 15 g) without skull fracture, consistent with heat-coagulated blood. Toxicological analysis detected triazolam and mirtazapine in the blood. Drowning was considered highly likely; however, definitive confirmation was limited by severe systemic thermal alteration. This case highlights a forensic diagnostic pitfall in which heat hematoma-like epidural changes following prolonged hot-water immersion may mimic traumatic epidural hematoma on PMCT, underscoring the importance of integrating scene information, toxicology, PMCT, and autopsy findings.

Triazole-containing compounds will result in the production of various pharmacologically effective drugs. Epilepsy is a severe neurological disorder primarily managed by controlling seizures using antiepileptic drugs, which are often related with side effects in rare circumstances and can be life-threatening. Triazolam and Alprazolam are both drugs containing a triazole ring, and these are well-established drugs that are used in the treatment of epilepsy. In the five-membered azole ring, triazole ring containing compounds consist of two carbon and three nitrogen atoms. Triazole, a heterocyclic compound, which contains at least two distinct elements in the ring that includes three nitrogen atoms and two carbon atoms as a part of the five-membered azole chain. The nitrogenous heterocyclic moiety triazole, which is commonly called pyrrodiazole, has the chemical formula C2H3N3. The triazole-containing derivatives with two different isomers are 1, 2, 3-triazoles and 1, 2, 4-triazoles. Triazole-containing compounds can be synthesized by various methods such as substitution reactions, ring opening, and rearrangement. Among these isomers of triazole, drugs containing 1,2,4-triazole possess different pharmacological properties such as antifungal, antineoplastic, antibacterial, antimycobacterial, and antiepileptic properties. 1,3,4-thiadiazole-linked 1,2,4-triazoles and 1,2,4-triazole-3-thione derivatives are important targets for potent anticonvulsant drug candidates in both clinical and preclinical studies. Many synthetic compounds have a structural characteristic of a 1, 2, 4-triazole ring with diverse therapeutic efficacy. 1,2,4-triazoles are a key scaffold in a large number of triazole derivatives that produce diverse pharmacological effects. This review focuses on the pharmacological significance and the synthesis of triazole derivatives in medicinal chemistry.

PURPOSE: We previously developed micro-segmental analysis (MSA), which measured detailed distributions of drugs in hair by segmenting a single hair strand at a 0.4-mm interval. MSA is effective in estimating the day of ingestion of hypnotics in drug-facilitated crimes. However, the disadvantage is that it needs considerable time to provide analytical results because hundreds of segments are analyzed using liquid chromatography (LC)-tandem mass spectrometry. In this study, the LC conditions were modified to improve the throughput of measurements. METHODS: Instrumental parts and parameters for LC were optimized to shorten the cycle time while maintaining separation between etizolam and triazolam, which have relatively similar retention times and the same nominal mass. Additionally, parallel column regeneration (PCR) available in two-dimensional LC system, which can switch channels through two identical columns, was used. RESULTS: The cycle time was reduced from 429 s to 235 s by changing the sample needle volume (from 15 to 30 µL), column length (from 150 to 50 mm) and injection volume (from 50 to 30 µL), and steepening the mobile phase gradient. Moreover, the use of PCR reduced the cycle time to 194 s by paralleling sample measurements with washing and equilibration of the lines. The effectiveness of the LC method modified using PCR was confirmed by the MSA of a long hair (978 segments) collected from a participant who had ingested hypnotics. CONCLUSIONS: This high-throughput measurement greatly contributed to saving machine time and early providing evidence of drug-facilitated crimes, showing detailed distributions of drugs in hair.

Triazolam (TRZ) is a representative benzodiazepine sedative-hypnotic drug that has gradually been abused due to the increasing societal pressures. To further provide a theoretical basis for the rationale use of TRZ and obtain more information for its metabolic process, in this study, human CYP3A4 was employed as the metabolic enzyme to investigate the metabolic mechanism of TRZ by multiple computational methods. Here, three types of substrate-binding conformations related to the diversity of TRZ metabolites are identified (pose A, pose B and pose C). The "sandwich" structure and the π-π stacking between TRZ and F304/porphyrin ring may be the key factors in dominating three substrate-binding conformations. Furthermore, we discovered pose A is the predominant binding mode, with Cα-H serving as the key metabolic site and CYP3A4-catalyzed Cα-H hydroxylation follows a hydrogen abstraction-rebound mechanism. More importantly, in hydroxylation process, the spin states of iron can regulate the metabolic reaction rate of TRZ and the highest rate of metabolism (5.96 s-1) is found in the quartet spin states. Based on our findings, it can be suggested that rational incorporating aromatic groups into TRZ could improve its metabolic stability. Meanwhile, the transition of the heme iron from a low-spin to a high-spin state appears to accelerate TRZ metabolism, potentially leading to the accumulation of α-OH triazolam in vivo, which may pose risks to human health. These results could enhance our understanding of CYP3A4-mediated regioselective metabolism of TRZ and provide a theoretical foundation and new perspective for studies on the metabolism of other triazole drugs.

Ensuring analyte stability is essential for accurate forensic and clinical detection of sedative-type drugs. This study systematically evaluated the stability of 22 sedative-type drugs and metabolites in human urine under controlled conditions varying by pH (4.0, 7.0), temperature (25°C, 4°C, -20°C), and freeze-thaw cycles (5 cycles), using a fully validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. While compounds such as midazolam, clobazam, and zolpidem remained highly stable, others-including alprazolam, triazolam, and lorazepam-exhibited notable degradation, particularly under acidic pH and elevated temperature. Flunitrazepam and clonazepam showed distinct degradation with the formation of 7-amino metabolites at neutral pH. Notably, this transformation occurred only in urine and not in phosphate-buffered saline, suggesting a urine-specific mechanism. These findings highlight the importance of compound-specific preservation strategies. In scenarios where analyte identity or sample pH cannot be verified promptly, immediate refrigeration or freezing (ideally at -20°C), along with minimizing freeze-thaw cycles, is strongly recommended to preserve sample integrity and ensure reliable toxicological interpretation.