ABSTRACT Alectinib is an oral anaplastic lymphoma kinase inhibitor, and M4 is its major pharmacologically active metabolite. Simultaneous quantification of alectinib and M4 in limited‐volume plasma is important for pediatric clinical studies, particularly when concentrations are low or exceed the calibration range. An LC–MS/MS method was developed and validated for simultaneous quantification of alectinib and its major metabolite, M4, in human plasma. Alectinib and M4 were extracted from 20 μL of plasma by protein precipitation and quantified using stable isotope‐labeled internal standards. Calibration ranges were 0.100–20.0 ng/mL for alectinib and 1.00–200 ng/mL for M4, with lower limits of quantification of 0.100 and 1.00 ng/mL, respectively. Full validation demonstrated acceptable selectivity, linearity, matrix effects, dilution reproducibility, stability, and carryover. Inter‐day accuracy (RE) ranged from −9.9% to 5.1%, and inter‐day precision (CV) was ≤ 8.3% for both analytes. Partial validation after analytical column replacement also met predefined acceptance criteria. The method was successfully applied to plasma samples obtained from one patient enrolled in an investigator‐initiated clinical trial of alectinib, with both alectinib and M4 remaining quantifiable up to 72 h after a single oral dose. This sensitive LC–MS/MS method supports the simultaneous quantification of alectinib and M4 in limited‐volume pediatric clinical study samples.
ABSTRACT A stability‐indicating reversed‐phase high‐performance liquid chromatographic method was developed and validated for quantitative determination of evogliptin in bulk drug substance and a marketed tablet formulation, followed by supporting liquid chromatography‐mass spectrometry investigation of selected stress‐associated components. Separation was achieved on an Eclipse XDB C8 column (250 × 4.6 mm, 5 μm) using acetonitrile and 0.1% formic acid in water (80:20, v/v) at 1.0 mL/min, with photodiode‐array detection at 268 nm and a 10 min run time. The method was linear over 10–60 μg/mL ( R 2 = 0.9993), with repeatability and intermediate precision below 2%. Mean recoveries ranged from 99.657 to 100.555%. The marketed tablet assay was 100.161 ± 0.333%. Forced‐degradation studies showed parent‐drug response losses of 10.90%, 23.61%, and 9.08% under acidic hydrolysis, alkaline hydrolysis, and oxidative stress conditions, respectively. Thermal and photolytic solid‐state exposure produced only 0.40% and 0.08% response loss, respectively, under the conditions investigated. Full‐scan LC–MS showed ions at m/z 186.2, 363.3, and 417.4 associated with DP1, DP2, and DP3, respectively. These findings support stress association but not definitive structures, as product‐ion MS/MS, accurate‐mass measurements, and authentic degradation‐product standards were not used. This study integrates quantitative stability‐indicating chromatographic assessment with conservative mass‐spectrometric investigation of selected stress‐associated components.
ABSTRACT This study combined systems pharmacology and computational biology to systematically investigate the multi‐target mechanisms and network features of Astilbin, a natural flavonoid glycoside, in Alzheimer's disease ( AD ) intervention. We collected potential targets of astilbin and AD ‐associated genes from PharmGKB, GeneCards, and OMIM databases and screened their intersecting targets. A PPI network was built via STRING, and hub genes were screened by topological analysis in Cytoscape. GO and KEGG enrichment analyses were performed, followed by molecular docking to assess binding affinity between Astilbin and core targets. In total, 250 overlapping targets were identified. Enrichment showed these targets participate in hormone/lipid responses, carboxylic acid metabolism and kinase activities. KEGG revealed Astilbin acts via PI3K‐Akt, MAPK and lipid metabolism pathways. Molecular docking verified that Astilbin exhibits low binding energy toward the core targets of Alzheimer's disease, implying a possible role in modulating neural signaling and ameliorating AD ‐related pathological processes. Collectively, Astilbin may contribute to AD intervention through a multi‐target, multi‐pathway network involving neural signaling, metabolism, inflammation, and cerebrovascular protection. This study clarifies the pharmacological mechanism of Astilbin and provides theoretical support for the development of natural product‐based therapeutic agents for neurodegenerative diseases.
ABSTRACT Yindan Pinggan capsule (YDPG) is a traditional Chinese formula for liver disorders, but its mechanism against nonalcoholic fatty liver disease (NAFLD) remains unclear. Here, UHPLC‐Q‐TOF‐MS was used to characterize YDPG's chemical profile, and the YDPG intestinal absorption solution (YDPGIAS) obtained via the everted gut sac model was analyzed to identify absorbable components. Network pharmacology, transcriptomic sequencing, molecular docking, and in vitro assays in oleic acid–induced BRL‐3A hepatocytes were integrated to predict targets and validate effects. A total of 164 compounds were identified from YDPG; 84 were detected in YDPGIAS, with 15 confirmed by reference standards. YDPGIAS significantly reduced lipid accumulation and lowered triglyceride, total cholesterol, LDL‐cholesterol, and HDL‐cholesterol levels. Integrated analysis identified the NF‐κB pathway as the core candidate mechanism, involving targets LYN, TNF, and IL1B, and key compounds including baicalin, ferulic acid, and gentiopicroside. Molecular docking predicted favorable binding affinities, and cellular experiments confirmed lipid‐lowering effects. These results demonstrate that YDPG ameliorates NAFLD through multicomponent, multitarget regulation of the NF‐κB‐mediated inflammatory pathway, providing a preliminary basis for further pharmacological and clinical investigation in NAFLD management.
ABSTRACT A rapid UPLC–MS/MS method was developed and validated for the simultaneous quantification of lodenafil (LDN) and its prodrug lodenafil carbonate (Helleva; LDC), a phosphodiesterase type 5 (PDE5) inhibitor developed by Cristália and approved by ANVISA for the treatment of erectile dysfunction in Brazil, in HLMs. Chromatographic separation was conducted utilizing Eclipse plus RP‐C18 (1.8 μm, 50 × 2.1 mm), under isocratic elution with 0.1% HCOOH in water and 0.1% HCOOH in ACN (30:70, v/v) at a 0.4 mL/min for 1.0 min. The method was linear from 1 to 4000 ng/mL ( R 2 = 0.9991; y = 0.2466x − 0.3277, 1/x weighting) for LDN and 1 to 4000 ng/mL ( R 2 = 0.9931; y = 5.5928x + 0.0311, 1/x weighting) for LDC. Substrate depletion of LDN in HLMs +NADPH yielded an in vitro t½ of 23.09 min and a Cl int of 35.11 mL/min/kg. LDC depletion in HLMs −NADPH yielded an in vitro t½ of 17.24 min and a Cl int of 47.02 mL/min/kg. LDC depletion proceeded at a comparable rate with and without NADPH. This finding is consistent with NADPH‐independent, esterase‐catalyzed hydrolysis of the carbonate linkage. Metabolic stability of the active moiety (LDN) and the depletion kinetics of the prodrug (LDC) were investigated.
