AAPS Journal最新文献

{"title":"Context-of-use-Guided Development and Validation of a Transthyretin Immunoassay: A Framework for Biomarker Assay Design.","authors":"Tanya Watanabe, John Canfield, Erin Anderson, Leighla Tayefeh, John L Allinson, Nicholas J Viney, Joel A Mathews","doi":"10.1208/s12248-026-01262-7","DOIUrl":"10.1208/s12248-026-01262-7","url":null,"abstract":"<p><p>Transthyretin (TTR) is a circulating protein biomarker central to the evaluation of therapies for transthyretin amyloidosis. Clinical trials with Eplontersen, a TTR-targeting antisense oligonucleotide, required an assay with a context of use differing from existing assay, requiring a more sensitive method. This study describes the development, optimization, and validation of a quantitative immunoassay for TTR, guided by a context-of-use framework. Method development focused on calibration curve modeling, weighting, parallelism, and determination of the minimal required dilution. Antibody selection and calibrator characterization followed established best practices. Pre-validation experiments optimized antibody concentrations and curve-fitting (five-parameter logistic with 1/y² weighting). Assay performance was then evaluated through validation, including assessments of accuracy, precision, parallelism, specificity, stability, interference of endogenous binding partners, and comparison against other commercially available assays. The assay demonstrated a functional lower limit of quantitation of approximately 0.07 mg/dL, which would enable measurement of > 95% reductions in TTR after treatment. Parallelism improved at higher dilutions. The assay showed robust performance across endogenous samples, recombinant TTR variants and in the presence of potential interferents. Total analytical variability was shown to be 29.4%. Comparisons with commercial assays showed strong correlation, though absolute values varied by platform. This work establishes a robust, context-driven assay for quantifying TTR in clinical studies. The development process highlights generalizable principles in biomarker assay design, including matrix considerations, variability analysis, and curve optimization. These insights provide a transferable framework for biomarker method development in regulated environments.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148145432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New Frontiers of Drug Development Through the Use of New Approach Methodologies.","authors":"M N Martinez, J P Mochel, K Allenspach, C Zdyrski, S Krishna, I Rusyn","doi":"10.1208/s12248-026-01263-6","DOIUrl":"10.1208/s12248-026-01263-6","url":null,"abstract":"<p><p>On November 2025, AAPS PharmSci 360 convened a symposium that included experts in the application of various types of New Approach Methodologies (NAMs). They shared their experiences and insights on topics that included microphysiological systems (MPS), the use of 3D organoids, and in silico tools. MPS systems were explored from the perspective of their testing and qualification through an academia-industry-government tissue chip testing consortium whose mission is to perform context-of-use-based testing of MPS and conduct comparative evaluations to support the use of MPS systems in safety assessments. 3D organoids were described and insights shared on how and when they may provide an appropriate tool for evaluating drug safety and effectiveness, and their use in supporting personalized medicine. Regarding in silico tools, examples were provided to describe their growing utility in predicting drug toxicity, population variability, and its potential benefits over traditional evidence-based toxicology. These tools include in silico models (e.g., physiologically based pharmacokinetic models), Artificial Intelligence (AI), and Machine Learning (ML) which, unlike other aspects of model informed drug development, reduce reliance on predefined models and allows for the integration of diverse sources of data. Computationally, these tools can generate predictions in hours where it would otherwise have taken weeks or months (and extensive experimentation). Within this meeting report, we highlight the key issues discussed during that symposium and share additional aspects to consider when developing a NAMs-based roadmap for specific contexts of use.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13334498/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148058201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to: Considerations in Kp,uu,brain-based Strategy for Selecting CNS-targeted Drug Candidates with Sufficient Target Coverage and Substantial Pharmacodynamic Effect.","authors":"Ling Zou, Huan-Chieh Chien, Devendra Pade, Yanfei Li, Minhkhoi Nguyen, Ravi Kanth Bhamidipati, Zhe Wang, Osatohanmwen Jessica Enogieru, Jan Wahlstrom","doi":"10.1208/s12248-026-01256-5","DOIUrl":"10.1208/s12248-026-01256-5","url":null,"abstract":"","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148058209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and Implementation of a Hinderance-based In Vitro Model for Porous Membranes.","authors":"Daniel A Paterson, Lucy Coleman, Yuri Dancik, Kanika Thakur, Yanling Zhang, Alice Maciel Tabosa, Sebastian Polak, James F Clarke","doi":"10.1208/s12248-026-01254-7","DOIUrl":"10.1208/s12248-026-01254-7","url":null,"abstract":"<p><p>In vitro release testing (IVRT) is an inexpensive and routine step in dermal formulation development where an inert membrane separates the donor formulation from a receptor solution. Experimental IVRT data provides insights into formulation behaviour in the absence of skin permeation complexities. This source of formulation data has the potential to be incorporated into a modelling workflow to aid parameterisation of a mechanistic dermal physiologically based pharmacokinetic (PBPK) model. Therefore, a hinderance-based IVRT model was developed within the Simcyp in vitro permeation testing (IVPT) module framework, using membrane characteristics to inform drug movement through inert, non-silicone, membranes. Modelling as a cylindrical pore system, the membranes key characteristics (pore size, porosity, tortuosity and membrane thickness) are utilised to parameterise a 2-step model where drug enters the membrane pores, diffuses across and partitions into the receptor solution. A description of the model's equations and setup are provided in addition to an ibuprofen in vitro case study using a range of membranes for model performance validation. Comparison of simulated IVRT to experimental data using membranes of microfiltration (pore diameters > 0.1 µm) resulted in acceptable performance, while simulation of nanofiltration membranes (pore diameters < 0.01 µm) require optimisation steps.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148044405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploration of Colonic Mucus as A Dissolution Compartment for Orally Administered Drugs.","authors":"Mingjun Wu, Eva M Karlsson, Christel A S Bergström","doi":"10.1208/s12248-026-01257-4","DOIUrl":"10.1208/s12248-026-01257-4","url":null,"abstract":"<p><p>Colonic mucus has gained interest as a dissolution compartment in the large intestine due to the large fraction of bound water. Notably, the mucus layer contains more water than what is freely mobile within the colonic lumen. Here, we studied solubility of a series of structurally and physicochemically diverse orally administered drugs. In total, 15 compounds were measured for their solubility in an artificial colonic mucus solution. Equilibrium solubility was measured using the traditional shake-flask method at 37°C in several important media for dissolution studies in the colon: the mucus solution, fed-state simulated colonic fluid (FeSSCoF) and the corresponding buffers. Molecular descriptors were generated by RDKit to identify the molecular features that might benefit from solubilization in the mucus solution. Of the studied compounds, 93% presented their highest solubility in the colonic mucus solution. Molecules with high lipophilicity were more likely to benefit from the solubilization potential of the mucus, herein identified to be a result of interactions with BSA and lipids in the system. To summarize, the large dataset investigated in this study clearly shows that the colonic mucus holds great potential as a dissolution compartment facilitating the drug absorption. Hence, the colonic mucus may drive absorption for poorly water-soluble compounds and should not only be viewed as a potential diffusion barrier for drug molecules and particles.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148007476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Mechanistic Framework Integrating Renal QSP-PK-PD and Machine Learning for Baseline-Informed Stratification of Diuretic Resistance.","authors":"Jialiang Zhou, Vingyou Lou, Yixin Zhao, Linxiu Tang, Zihan Hu, Jiaqi Sun, Jun Chen, Peng Cheng, Yu Cheng, Hua He","doi":"10.1208/s12248-026-01255-6","DOIUrl":"10.1208/s12248-026-01255-6","url":null,"abstract":"<p><p>Diuretic resistance represents a major source of heterogeneity in loop diuretic response and remains a key barrier to effective decongestion in heart failure. A key clinical challenge is the early identification of patients at high risk of an inadequate response to standard-dose furosemide in order to inform timely treatment intensification or alternative decongestive strategies. However, current approaches remain reactive and rely on post-treatment response, limiting prospective risk stratification and treatment decision-making. To address this problem, we integrated a mechanistic renal quantitative systems pharmacology (QSP) model with furosemide pharmacokinetics and pharmacodynamics to enable baseline-informed risk stratification of diuretic resistance. The model was calibrated using published clinical pharmacokinetic and renal response data and applied to generate a physiologically constrained virtual patient population capturing heterogeneity in renal function, tubular sodium handling, and neurohormonal activation. Machine learning methods were incorporated as complementary analytical tools to identify and validate baseline physiological determinants that define risk categories of diuretic response within the mechanistic simulation framework. Model-based analyses indicated that diuretic resistance arises from the combined effects of reduced filtration capacity, enhanced tubular sodium avidity, diminished pharmacodynamic sensitivity and sustained neurohormonal activation. Across analytical approaches, baseline fractional excretion of sodium and glomerular filtration rate emerged as integrative biomarkers that stratify patients into distinct risk categories of diuretic response, reflecting a mechanism-informed reduction of underlying physiological variability. These findings provide a mechanistically grounded framework for baseline-informed risk stratification and may inform earlier identification of patients at risk of inadequate response, supporting model-informed evaluation of treatment intensification or alternative decongestive strategies.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148007411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HLA Class II Alleles DRB1*11:01 and DQB1*03:01 Unmask Immunogenetic Susceptibility to Anti-Nivolumab Antibodies in Combination with Ipilimumab.","authors":"Surendran Rajendran, Andres H Gutierrez, Ming-Shan Chien, Tracy Tang, Jochem Gokemeijer, Daron Forman, Anne S De Groot, Vibha Jawa, Lora Hamuro","doi":"10.1208/s12248-026-01244-9","DOIUrl":"10.1208/s12248-026-01244-9","url":null,"abstract":"<p><p>Biologic combinations that modulate immune responses are increasingly used in oncology, enhancing anti-tumor immunity but also raising the risk of anti-drug antibody (ADA) development. Nivolumab administered in combination with ipilimumab results in significantly higher nivolumab ADA incidence (44%) compared to nivolumab monotherapy (12%). While previous analyses showed no clinical impact of nivolumab ADA on safety and efficacy for the combination, it provided a unique dataset to evaluate genetic predispositions to nivolumab ADA development and impact of co-administered ipilimumab. HLA Class II alleles were evaluated for associations with nivolumab ADA. Logistic regression analyses were performed on HLA-DRB1, DQB1, and DQA1 alleles from melanoma subjects treated with nivolumab plus ipilimumab and nivolumab alone. Univariate screening identified candidate alleles that were associated with immunogenicity. This was followed by multivariable modeling to assess independent and combined effects after accounting for sex, age and region. Multivariable analyses indicated that DRB1*11:01 (Odds Ratio (OR) = 2.5, p-value (p) = 0.011) and DQB1*03:01 (OR = 3.2, p = 0.0002) were each significantly associated with nivolumab ADA positivity in combination therapy. The combined presence of both alleles conferred an even stronger association (OR = 4.1, p = 0.0017). No significant associations were observed for nivolumab monotherapy. In silico predictions and MHC-associated peptide proteomics (MAPPs) confirmed high-affinity binding of a nivolumab framework peptide (a known Treg epitope) to DRB1*11:01. DRB1*11:01 and DQB1*03:01 may predict anti-nivolumab antibody development in the context of combination therapy. These findings underscore the role of host genetics and immune modulation by ipilimumab in shaping immunogenicity.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147984469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunogenicity Assessment for siRNA Therapeutics: A Risk-Based Proposal for Event-Driven Testing.","authors":"An Zhao, Sarah Bond, Valerie Clausen, Mu Chen, Ching-Ha Lai, Joshua Zylstra, Christine Grimaldi, Michael A Partridge","doi":"10.1208/s12248-026-01260-9","DOIUrl":"10.1208/s12248-026-01260-9","url":null,"abstract":"<p><p>Oligonucleotide therapeutics (ONTs) have emerged as a transformative treatment modality with numerous products approved globally for addressing a variety of diseases. Within this class, siRNAs therapeutics have achieved remarkable success, with eight approvals to date. siRNA therapeutics are short double-stranded RNAs designed to hybridize with complementary mRNA sequences and regulate disease-related protein expression through endogenous RNA interference (RNAi) mechanism. This manuscript outlines a framework of immunogenicity risk assessment for siRNA therapeutics, discusses the challenges associated with anti-drug antibody (ADA) assay development, and reviews available immunogenicity data from both approved and investigational GalNAc-siRNA therapeutics. Across clinical development programs, GalNAc- conjugated siRNA therapeutics have demonstrated a low immunogenicity risk profile, supported by comprehensive immunogenicity risk assessment and accumulated clinical data. Reported treatment-emergent ADA response has been generally low (≤ 6%), with no observed impact on pharmacokinetics (PK), pharmacodynamics (PD), efficacy and/or safety. Given the favorable immunogenicity profile observed to date, routine prospective ADA assessments throughout clinical development may not be scientifically warranted for all siRNA therapeutics. Instead, the manuscript advocates adopting a risk-based immunogenicity assessment strategy. This approach emphasizes immunogenicity risk assessment early in the program development, for low-risk molecules, such as GalNAc- or lipid-conjugated siRNA, ADA samples can be collected and banked during early phase studies for retrospective testing if evidence emerges of altered PK, PD, or immune-mediated adverse events. This risk-based immunogenicity assessment strategy ensures patient safety and therapeutic efficacy while streamlining development process for siRNA therapeutics.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147976927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanotechnology-enhanced Natural Products for Cancer Chemoprevention: Molecular Mechanisms and Clinical Translation.","authors":"Rajat Nath, Sibashish Kityania, Deepa Nath, Jayanta Kumar Patra, Emran Habibi, Satyajit D Sarker, Anupam Das Talukdar, Lutfun Nahar","doi":"10.1208/s12248-026-01258-3","DOIUrl":"10.1208/s12248-026-01258-3","url":null,"abstract":"<p><p>Cancer remains a major global health challenge. Natural compounds, such as curcumin, resveratrol, genistein, thymoquinone, and paclitaxel, show chemopreventive activity by modulating signaling pathways, including PI3K/Akt, NF-κB, and p53. These agents also promote apoptosis, autophagy, and DNA repair. However, their clinical use is restricted by poor solubility, instability, and low bioavailability. Nanotechnology offers solutions by improving stability, enhancing pharmacokinetics, and enabling targeted delivery. Liposomes, polymeric nanoparticles, dendrimers, and albumin-bound systems amplify the anticancer effects of natural compounds. Preclinical studies confirm improved efficacy, while early clinical trials reveal both promise and barriers. The key translational challenges include immune clearance, large-scale reproducibility, and regulatory approval. This review highlights the synergy between nanotechnology and natural compounds in cancer chemoprevention and outlines opportunities for future research.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147935152","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In-depth Characterization of Photo-Stressed Antibody-Drug Conjugates by Mass Spectrometry.","authors":"Xiao Pan, Yimeng Zhao, Shukun Luo, Joshua Bulos, Jieqiang Zhong, Yuetian Yan, Zhijie Wu, Yue Su, Sisi Zhang, Shunhai Wang, Hui Xiao, Wenhua Wang, Dingjiang Liu, Ning Li, Haibo Qiu","doi":"10.1208/s12248-026-01247-6","DOIUrl":"10.1208/s12248-026-01247-6","url":null,"abstract":"<p><p>Camptothecin (CPT)-based antibody-drug conjugates (ADCs) have become popular oncology biotherapeutics that integrate the targeted specificity of monoclonal antibodies (mAbs) with camptothecin and its derivatives. Because mAbs are vulnerable to photodegradation and the light-absorbing characteristics of camptothecin, understanding the effects of light exposure on CPT-based ADCs is critical to ensure product quality, efficacy, and safety. Herein, we harnessed the capabilities of advanced liquid chromatography-mass spectrometry (LC-MS) to perform in-depth molecular-level characterization of photo-stressed antibodies conjugated with a camptothecin derivative payload, deruxtecan (DXd). Peptide mapping via LC-MS/MS revealed significant oxidative modifications in methionine, tryptophan, and histidine residues, and degradation of the linker payload in the photo-stressed ADC molecules. Native SEC-MS with post-column denaturation was conducted to further characterize the covalent nature of high molecular weight species formed under photo stress and provide insights into aggregation mechanisms. Additionally, coupling LC to high-resolution MS enabled the identification and quantitation of degradants from excipients and the linker payload, thus providing a comprehensive understanding of light exposure effects on DXd ADCs. Application of these LC-MS based assays to evaluate the effects of various formulations on photostability revealed that histidine and methionine act as oxidative scavengers that protect ADC molecules against light-induced degradation. This study highlights the utility of LC-MS methods in the comprehensive characterization of photo-stressed CPT-based ADCs, and provides valuable insights for process optimization, formulation development, and storage condition recommendations.</p>","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"28 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2026-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147876392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}