AAPS Journal最新文献_第7页

Characterization and Applications of Permeabilized Hepatocytes in Drug Discovery. 药物发现中渗透肝细胞的表征和应用。

IF 5 3区医学

AAPS Journal Pub Date : 2024-03-28 DOI: 10.1208/s12248-024-00907-9

Sam Zhang, Christine C Orozco, Lloyd Wei Tat Tang, Jillian Racich, Anthony A Carlo, George Chang, David Tess, Christopher Keefer, Li Di

{"title":"Characterization and Applications of Permeabilized Hepatocytes in Drug Discovery.","authors":"Sam Zhang, Christine C Orozco, Lloyd Wei Tat Tang, Jillian Racich, Anthony A Carlo, George Chang, David Tess, Christopher Keefer, Li Di","doi":"10.1208/s12248-024-00907-9","DOIUrl":"10.1208/s12248-024-00907-9","url":null,"abstract":"Hepatocytes are one of the most physiologically relevant in vitro liver systems for human translation of clearance and drug-drug interactions (DDI). However, the cell membranes of hepatocytes can limit the entry of certain compounds into the cells for metabolism and DDI. Passive permeability through hepatocytes can be different in vitro and in vivo, which complicates the human translation. Permeabilized hepatocytes offer a useful tool to probe mechanistic understanding of permeability-limited metabolism and DDI. Incubation with saponin of 0.01% at 0.5 million cells/mL and 0.05% at 5 million cells/mL for 5 min at 37°C completely permeabilized the plasma membrane of hepatocytes, while leaving the membranes of subcellular organelles intact. Permeabilized hepatocytes maintained similar enzymatic activity as intact unpermeabilized hepatocytes and can be stored at -80°C for at least 7 months. This approach reduces costs by preserving leftover hepatocytes. The relatively low levels of saponin in permeabilized hepatocytes had no significant impact on the enzymatic activity. As the cytosolic contents leak out from permeabilized hepatocytes, cofactors need to be added to enable metabolic reactions. Cytosolic enzymes will no longer be present if the media are removed after cells are permeabilized. Hence permeabilized hepatocytes with and without media removal may potentially enable reaction phenotyping of cytosolic enzymes. Although permeabilized hepatocytes work similarly as human liver microsomes and S9 fractions experimentally requiring addition of cofactors, they behave more like hepatocytes maintaining enzymatic activities for over 4 h. Permeabilized hepatocytes are a great addition to the drug metabolism toolbox to provide mechanistic insights.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 3","pages":"38"},"PeriodicalIF":5.0,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140319786","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}

引用次数: 0

Novel Multiplexed High Throughput Screening of Selective Inhibitors for Drug-Metabolizing Enzymes Using Human Hepatocytes. 利用人体肝细胞进行药物代谢酶选择性抑制剂的新型多重高通量筛选

IF 5 3区医学

AAPS Journal Pub Date : 2024-03-28 DOI: 10.1208/s12248-024-00908-8

Jianhua Liu, Daria Vernikovskaya, Gary Bora, Anthony Carlo, Woodrow Burchett, Samantha Jordan, Lloyd Wei Tat Tang, Joy Yang, Ye Che, George Chang, Matthew D Troutman, Li Di

{"title":"Novel Multiplexed High Throughput Screening of Selective Inhibitors for Drug-Metabolizing Enzymes Using Human Hepatocytes.","authors":"Jianhua Liu, Daria Vernikovskaya, Gary Bora, Anthony Carlo, Woodrow Burchett, Samantha Jordan, Lloyd Wei Tat Tang, Joy Yang, Ye Che, George Chang, Matthew D Troutman, Li Di","doi":"10.1208/s12248-024-00908-8","DOIUrl":"10.1208/s12248-024-00908-8","url":null,"abstract":"Selective chemical inhibitors are critical for reaction phenotyping to identify drug-metabolizing enzymes that are involved in the elimination of drug candidates. Although relatively selective inhibitors are available for the major cytochrome P450 enzymes (CYP), they are quite limited for the less common CYPs and non-CYPs. To address this gap, we developed a multiplexed high throughput screening (HTS) assay using 20 substrate reactions of multiple enzymes to simultaneously monitor the inhibition of enzymes in a 384-well format. Four 384-well assay plates can be run at the same time to maximize throughput. This is the first multiplexed HTS assay for drug-metabolizing enzymes reported. The HTS assay is technologically enabled with state-of-the-art robotic systems and highly sensitive modern LC-MS/MS instrumentation. Virtual screening is utilized to identify inhibitors for HTS based on known inhibitors and enzyme structures. Screening of ~4600 compounds generated many hits for many drug-metabolizing enzymes including the two time-dependent and selective aldehyde oxidase inhibitors, erlotinib and dibenzothiophene. The hit rate is much higher than that for the traditional HTS for biological targets due to the promiscuous nature of the drug-metabolizing enzymes and the biased compound selection process. Future efforts will focus on using this method to identify selective inhibitors for enzymes that do not currently have quality hits and thoroughly characterizing the newly identified selective inhibitors from our screen. We encourage colleagues from other organizations to explore their proprietary libraries using a similar approach to identify better inhibitors that can be used across the industry.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 3","pages":"36"},"PeriodicalIF":5.0,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140319788","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}

引用次数: 0

Development and Validation of a Cell-Based Binding Neutralizing Antibody Assay for an Antibody-Drug Conjugate. 开发并验证基于细胞的抗体药物共轭物结合中和抗体检测方法

IF 5 3区医学

AAPS Journal Pub Date : 2024-03-28 DOI: 10.1208/s12248-024-00909-7

Weifeng Xu, Nazneen Bano, Olguitza Guzman-Valdes, Jessica Amberman, Elisha Bandlamudi, Pooja Khanna, Rebecca Carmean, Roy Helmy

{"title":"Development and Validation of a Cell-Based Binding Neutralizing Antibody Assay for an Antibody-Drug Conjugate.","authors":"Weifeng Xu, Nazneen Bano, Olguitza Guzman-Valdes, Jessica Amberman, Elisha Bandlamudi, Pooja Khanna, Rebecca Carmean, Roy Helmy","doi":"10.1208/s12248-024-00909-7","DOIUrl":"10.1208/s12248-024-00909-7","url":null,"abstract":"The utilization of antibody-drug conjugates (ADCs) has gained considerable attention in the field of targeted cancer therapy due to their ability to synergistically combine the specificity of monoclonal antibodies (mAbs) and the potency of small molecular drugs. However, the immunogenic nature of the antibody component within ADCs warrants the need for robust immunogenicity testing, including a neutralizing antibody (NAb) assay. Since the mechanism of action (MOA) of the ADC is to first bind to the target cells and then release the payload intracellularly to kill the cells, the most relevant NAb assay format would be a cell-based killing assay. However, in this paper, we present a case where a cell-based killing assay could not be developed after multiple cell lines and NAb-positive controls (PC) had been tested. Surprisingly, contrary to our expectations, all NAb PCs tested exhibited an increased killing effect on the target cells, instead of the expected protective response. This unexpected phenomenon most likely is due to the non-specific internalization of drug/NAb complexes via FcγRs, as an excessive amount of human IgG1 and mouse IgG2a, but not mouse IgG1, greatly inhibited drug or drug/NAb complexes induced cell death. To overcome this obstacle, we implemented a novel cell-based binding assay utilizing the Meso Scale Discovery (MSD) platform. We also propose that an in vitro cell killing NAb assay is limited to at best monitoring the target binding and internalization induced cell death, but not by-stander killing induced by prematurely released or dead-cell released payload, hence cannot really mimic the in vivo MOA of ADC.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 3","pages":"37"},"PeriodicalIF":5.0,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140319787","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}

引用次数: 0

State of the Art of Silica Nanoparticles: An Overview on Biodistribution and Preclinical Toxicity Studies. 二氧化硅纳米粒子的技术现状：生物分布和临床前毒性研究概述。

IF 5 3区医学

AAPS Journal Pub Date : 2024-03-21 DOI: 10.1208/s12248-024-00906-w

Joshua Yu, Nirnoy Dan, Seyyed Majid Eslami, Xiuling Lu

引用次数: 0

Science- and Risk-Based Stability Strategies to Support Product Lifecycle Changes. 以科学和风险为基础的稳定性战略，支持产品生命周期的变化。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-03-14 DOI: 10.1208/s12248-024-00903-z

Lori McCaig, Steven Nowak, Alexander Abbott, Jenny Carhart, Megan E McMahon, Elke Debie, Hanlin Li, Francis Maina, Andrea J Ji, Mingkun Fu, Yan Wu, Andrew Lennard, Tony Mazzeo, Chad Wolfe, Robert Timpano, Yelizaveta Babayan, Lars Gruenig

{"title":"Science- and Risk-Based Stability Strategies to Support Product Lifecycle Changes.","authors":"Lori McCaig, Steven Nowak, Alexander Abbott, Jenny Carhart, Megan E McMahon, Elke Debie, Hanlin Li, Francis Maina, Andrea J Ji, Mingkun Fu, Yan Wu, Andrew Lennard, Tony Mazzeo, Chad Wolfe, Robert Timpano, Yelizaveta Babayan, Lars Gruenig","doi":"10.1208/s12248-024-00903-z","DOIUrl":"10.1208/s12248-024-00903-z","url":null,"abstract":"ICH Q12 asserts that science- and risk-based approaches are applicable to stability studies supporting Chemistry, Manufacturing and Controls (CMC) post-approval changes (PAC) to enable more timely implementation; however, no guidance or specific examples are provided to demonstrate how prior knowledge of the product can inform the risk assessment for the proposed change(s). Ten diverse case studies are presented in this manuscript to demonstrate how science- and risk-based stability strategies were used to support drug substance and product CMC PAC and lifecycle management activities. The accumulated stability knowledge held by original manufacturers of marketed products is substantial, and different elements of this knowledge base were used to assess the risks and impact of the proposed changes for confident change management. This paper provides ways to leverage science- and risk-based stability strategies as part of the post-approval change-management risk-mitigation strategy, which may enable a reduced stability data commitment and/or a reduced reporting category for change implementation.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 2","pages":"34"},"PeriodicalIF":4.5,"publicationDate":"2024-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140133150","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}

引用次数: 0

Quality by Design Framework Applied to GMMA Purification. 质量源于设计框架应用于 GMMA 净化。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-03-08 DOI: 10.1208/s12248-024-00902-0

Carlo Giannelli, Francesca Necchi, Elena Palmieri, Davide Oldrini, Beatrice Ricchetti, Maria M Papathanasiou, Zoltan Kis, Cleo Kontoravdi, Cristiana Campa, Francesca Micoli

引用次数: 0

Implications of Immunogenicity Testing for Therapeutic Monoclonal Antibodies: A Quantitative Pharmacology Framework. 治疗性单克隆抗体免疫原性测试的意义：定量药理学框架。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-03-07 DOI: 10.1208/s12248-024-00901-1

Jason H Williams, Kai H Liao, Donghua Yin, Xu Meng

{"title":"Implications of Immunogenicity Testing for Therapeutic Monoclonal Antibodies: A Quantitative Pharmacology Framework.","authors":"Jason H Williams, Kai H Liao, Donghua Yin, Xu Meng","doi":"10.1208/s12248-024-00901-1","DOIUrl":"10.1208/s12248-024-00901-1","url":null,"abstract":"The interpretation of immunogenicity results for a mAb product and prediction of its clinical consequences remain difficult, despite enormous advances in methodologies and efforts toward the best practice for consistent data generation and reporting. To this end, the contribution from the clinical pharmacology discipline has been largely limited to comparing descriptively the pharmacokinetic (PK) profiles by antidrug antibodies (ADA) status or testing the significance of ADA as a covariate in a population PK setting, similar to the practice for small-molecule drugs in investigating the effect of an intrinsic/extrinsic factor on the drug disposition. There is a need for a mAb disposition framework that captures the dynamics of ADA formation and drug's interactions with the ADA and target as parts of the drug distribution and elimination. Here we describe such a framework and examine it against the PK, ADA, and clinical response data from a phase 3 trial in patients treated with adalimumab. The proposed framework offered a generalized understanding of how the dose, target affinity, and drug/ADA analyte forms affects the manifestation of ADA response with regard to its detections and alterations of drug disposition and effectiveness. Furthermore, as an example, its utility for dose considerations was demonstrated through predicting for late-stage trials of a PCSK9 inhibitor in terms of development in ADA incidence and titers, and consequences on the drug disposition, interaction with target, and downstream lowering effect on LDL-C.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 2","pages":"31"},"PeriodicalIF":4.5,"publicationDate":"2024-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140061156","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}

引用次数: 0

A Short Update on the Use of Monoclonal Antibodies in COVID-19. 关于在 COVID-19 中使用单克隆抗体的简短更新。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-03-05 DOI: 10.1208/s12248-024-00904-y

Antonio Vitiello, Michela Sabbatucci, Annarita Ponzo, Antonio Salzano, Andrea Zovi

引用次数: 0

Investigation of Antibody Pharmacokinetics in the Brain Following Intra-CNS Administration and Development of PBPK Model to Characterize the Data. 中枢神经系统内给药后脑内抗体药代动力学的研究，以及为描述数据而开发的 PBPK 模型。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-03-05 DOI: 10.1208/s12248-024-00898-7

Shengjia Wu, Hsueh-Yuan Chang, Ekram Ahmed Chowdhury, Hsien Wei Huang, Dhaval K Shah

{"title":"Investigation of Antibody Pharmacokinetics in the Brain Following Intra-CNS Administration and Development of PBPK Model to Characterize the Data.","authors":"Shengjia Wu, Hsueh-Yuan Chang, Ekram Ahmed Chowdhury, Hsien Wei Huang, Dhaval K Shah","doi":"10.1208/s12248-024-00898-7","DOIUrl":"10.1208/s12248-024-00898-7","url":null,"abstract":"Despite the promising potential of direct central nervous system (CNS) antibody administration to enhance brain exposure, there remains a significant gap in understanding the disposition of antibodies following different intra-CNS injection routes. To bridge this knowledge gap, this study quantitatively investigated the brain pharmacokinetics (PK) of antibodies following intra-CNS administration. The microdialysis samples from the striatum (ST), cerebrospinal fluid (CSF) samples through cisterna magna (CM) puncture, plasma, and brain homogenate samples were collected to characterize the pharmacokinetics (PK) profiles of a non-targeting antibody, trastuzumab, following intracerebroventricular (ICV), intracisternal (ICM), and intrastriatal (IST) administration. For a comprehensive analysis, these intra-CNS injection datasets were juxtaposed against our previously acquired intravenous (IV) injection data obtained under analogous experimental conditions. Our findings highlighted that direct CSF injections, either through ICV or ICM, resulted in ~ 5-6-fold higher interstitial fluid (ISF) drug exposure than IV administration. Additionally, the low bioavailability observed following IST administration indicates the existence of a local degradation process for antibody elimination in the brain ISF along with the ISF bulk flow. The study further refined a physiologically based pharmacokinetic (PBPK) model based on new observations by adding the perivascular compartments, oscillated CSF flow, and the nonspecific uptake and degradation of antibodies by brain parenchymal cells. The updated model can well characterize the antibody PK following systemic and intra-CNS administration. Thus, our research offers quantitative insight into antibody brain disposition pathways and paves the way for determining optimal dosing and administration strategies for antibodies targeting CNS disorders.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 2","pages":"29"},"PeriodicalIF":4.5,"publicationDate":"2024-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140040823","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}

引用次数: 0

The Role of Model Master Files for Sharing, Acceptance, and Communication with FDA. 示范主文件在与 FDA 共享、接受和交流方面的作用。

IF 4.5 3区医学

AAPS Journal Pub Date : 2024-02-27 DOI: 10.1208/s12248-024-00897-8

Lanyan Fang, Yuqing Gong, Andrew C Hooker, Viera Lukacova, Amin Rostami-Hodjegan, Mark Sale, Stella Grosser, Rebeka Jereb, Rada Savic, Carl Peck, Liang Zhao

{"title":"The Role of Model Master Files for Sharing, Acceptance, and Communication with FDA.","authors":"Lanyan Fang, Yuqing Gong, Andrew C Hooker, Viera Lukacova, Amin Rostami-Hodjegan, Mark Sale, Stella Grosser, Rebeka Jereb, Rada Savic, Carl Peck, Liang Zhao","doi":"10.1208/s12248-024-00897-8","DOIUrl":"10.1208/s12248-024-00897-8","url":null,"abstract":"With the evolving role of Model Integrated Evidence (MIE) in generic drug development and regulatory applications, the need for improving Model Sharing, Acceptance, and Communication with the FDA is warranted. Model Master File (MMF) refers to a quantitative model or a modeling platform that has undergone sufficient model Verification & Validation to be recognized as sharable intellectual property that is acceptable for regulatory purposes. MMF provides a framework for regulatorily acceptable modeling practice, which can be used with confidence to support MIE by both the industry and the U.S. Food and Drug Administration (FDA). In 2022, the FDA and the Center for Research on Complex Generics (CRCG) hosted a virtual public workshop to discuss the best practices for utilizing modeling approaches to support generic product development. This report summarizes the presentations and panel discussions of the workshop symposium entitled \"Model Sharing, Acceptance, and Communication with the FDA\". The symposium and this report serve as a kick-off discussion for further utilities of MMF and best practices of utilizing MMF in drug development and regulatory submissions. The potential advantages of MMFs have garnered acknowledgment from model developers, industries, and the FDA throughout the workshop. To foster a unified comprehension of MMFs and establish best practices for their application, further dialogue and cooperation among stakeholders are imperative. To this end, a subsequent workshop is scheduled for May 2-3, 2024, in Rockville, Maryland, aiming to delve into the practical facets and best practices of MMFs pertinent to regulatory submissions involving modeling and simulation methodologies.","PeriodicalId":50934,"journal":{"name":"AAPS Journal","volume":"26 2","pages":"28"},"PeriodicalIF":4.5,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139984449","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}

引用次数: 0