Pharmaceutical Research最新文献

{"title":"Synthesis and Biological Evaluation of Novel Triazine Analogs as Rad6 Inhibitors.","authors":"Qian Lin, Ambikai Gajan, Ignatius Nguyen, Shiv Sharma, Pratima Nangia-Makker, Steven Firestine, Malathy P Shekhar","doi":"10.1007/s11095-025-03838-y","DOIUrl":"10.1007/s11095-025-03838-y","url":null,"abstract":"<p><p>Rad6 is an E2 ubiquitin-conjugating enzyme that plays critical roles in genome maintenance and proteostasis. Rad6 is frequently overexpressed in many cancers and promotes cancer development, progression, and chemotherapy resistance.</p><p><strong>Purpose: </strong>Given its role in cancer development and progression, Rad6 is an underexplored therapeutic target. Previous research identified compound SMI#9 as a small molecule inhibitor of Rad6. Despite its potency, SMI#9 has limited efficacy in vivo due to its limiting water solubility and the presence of a labile ester group.</p><p><strong>Methods: </strong>To address these limitations, we prepared a series of SMI#9 analogs in which the ester group was replaced with a secondary amine, and their effects on Rad6B-mediated ubiquitination of histone H2A were evaluated. In vivo interaction with Rad6 was assessed using cellular thermal shift assays. SMI#9 analog effects on cell survival and migration of triple negative and endocrine-resistant breast cancer, and melanoma cells were measured using MTT and Boyden chamber assays. Autophagy, mitochondrial function, and β-catenin localization were measured using CytoID, Mitotracker, and immunostaining, respectively. Cellular uptakes of analogs were determined by mass spectroscopy.</p><p><strong>Results: </strong>Analogs #4 and #6 inhibited H2A ubiquitination, induced autophagy and mitochondrial dysfunction, downregulated intracellular β-catenin, and inhibited proliferation. #6 targets Rad6 in vivo. #4 and #6 are chemically related, and #4 undergoes in vivo conversion to #6.</p><p><strong>Conclusions: </strong>#6 retains all the properties of SMI#9 but with lesser potency. However, its improved water solubility and metabolic stability allows for in vivo studies that were previously precluded due to the poor physicochemical properties of SMI#9.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"511-527"},"PeriodicalIF":3.5,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531010","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":"Structure Bioinformatics of Six Human Integral Transmembrane Enzymes and their AlphaFold3 Predicted Water-Soluble QTY Analogs: Insights into FACE1 and STEA4 Binding Mechanisms.","authors":"Edward Chen, Emily Pan, Shuguang Zhang","doi":"10.1007/s11095-025-03822-6","DOIUrl":"10.1007/s11095-025-03822-6","url":null,"abstract":"<p><strong>Objective: </strong>Human integral membrane enzymes are essential for catalyzing a wide range of biochemical reactions and regulating key cellular processes. However, studying these enzymes remains challenging due to their hydrophobic nature, which necessitates the use of detergents. This study explores whether applying the QTY code can reduce the hydrophobicity of these enzymes while preserving their structures and functions, thus facilitating bioinformatics analysis of six key integral membrane enzymes: MGST2, LTC4S, PTGES, FACE1, STEA4, and SCD.</p><p><strong>Methods: </strong>The water-soluble QTY analogs of the six membrane enzymes were predicted using AlphaFold3. The predicted structures were superposed with CyroEM determined native structures in PyMOL to observe changes in structure and protein-ligand binding ability.</p><p><strong>Results: </strong>The native membrane enzymes superposed well with their respective QTY analogs, with the root mean square deviation (RMSD) ranging from 0.273 Å to 0.875 Å. Surface hydrophobic patches on the QTY analogs were significantly reduced. Importantly, the protein-ligand interactions in FACE1 and STEA4 were largely preserved, indicating maintained functionality.</p><p><strong>Conclusion: </strong>Our structural bioinformatics studies using the QTY code and AlphaFold3 not only provide the opportunities of designing more water-soluble integral membrane enzymes, but also use these water-soluble QTY analogs as antigens for therapeutic monoclonal antibody discovery to specifically target the key integral membrane enzymes.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"291-305"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880043/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143449910","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":"Theranostic Near-Infrared Monoamine Oxidase Inhibitor (NMI) Protein Binding Interactions with MAOA and Albumin.","authors":"Ronald W Irwin, Unnati H Shah, Shivani Soni, Heinz Josef Lenz, Jean C Shih","doi":"10.1007/s11095-025-03827-1","DOIUrl":"10.1007/s11095-025-03827-1","url":null,"abstract":"<p><strong>Purpose: </strong>The protein binding interactions of near-infrared monoamine oxidase inhibitor (NMI) are reported here.</p><p><strong>Methods: </strong>NMI-bound proteins were examined by fluorescent SDS-PAGE and mass spectrometry using tumor tissues from brain and colon cancer mouse models.</p><p><strong>Results: </strong>This study shows protein interactions with NMI, a chemical conjugate of MAOA inhibitor clorgyline and tumor-seeking dye, MHI-148. NMI fluorescence in MAOA knock-out (KO) mice was significantly lower compared to WT mice, including whole animal, organs, and tissue lysates which indicated that NMI binds to MAOA. Pure recombinant MAOA protein was detectable as a single fluorescent band that migrated at ~ 65kD. NMI inhibited MAOA activity (IC₅₀ 1-5 µM). In a glioma mouse model, NMI targeted specifically to tumor with high contrast to adjacent normal brain, shown by a 65 kD protein band. Recent studies demonstrated heptamethine cyanine dyes (e.g., MHI-148) interact with serum albumin, contributing to tumor uptake and cancer cell internalization. Our study shows NMI binds to albumin but highly prefers MAOA, providing a plausible mechanism for systemic drug delivery via serum albumin to the tumor target and subsequent MAOA inhibition. Further studies in a colon cancer mouse model found the ~ 65 kD SDS-PAGE band, bound to NMI, contained both MAOA and albumin proteins by mass spectrometry.</p><p><strong>Conclusion: </strong>NMI was shown to interact with MAOA and the blood carrier protein, albumin. This study provides insights for drug delivery and protein target specificity of NMI to image and treat cancer.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"307-318"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880180/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143190123","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":"Dietary phytochemical indole-3-carbinol regulates metabolic reprogramming in mouse prostate tissue.","authors":"Rebecca Mary Peter, Md Shahid Sarwar, Lujing Wang, Pochung Chou, Chao Wang, Yujue Wang, Xiaoyang Su, Ah-Ng Kong","doi":"10.1007/s11095-025-03820-8","DOIUrl":"10.1007/s11095-025-03820-8","url":null,"abstract":"<p><strong>Purpose: </strong>Indole-3-carbinol (I3C) is shown to possess multiple pharmacological activities such as anti-inflammatory, antimicrobial, antioxidant, antiviral, and anti-cancer activities. It is widely accepted as modulator of multiple signaling pathways particularly those related to cell cycle, cell growth and division, angiogenesis, apoptosis and immunity. We explored the metabolic reprogramming based on treatment with I3C in mice prostate tissue.</p><p><strong>Methods: </strong>In this study we utilized Pten knockout (KO)-induced prostate tumorigenesis mouse model to examine mechanism of action of I3C via metabolic rewiring. Phosphatase and tensin homolog deleted on chromosome 10 (Pten), a tumor suppressor gene is frequently found to be mutated or deleted in prostate cancer. Untargeted metabolomics was performed using liquid-chromatography mass-spectrometry (LC-MS) based platform to investigate Pten-dependent and Pten-independent metabolic targets of I3C.</p><p><strong>Results: </strong>The most impacted pathways by I3C included pyrimidine metabolism, arginine and proline metabolism, porphyrin metabolism, citrate cycle and lipoic acid metabolism.</p><p><strong>Conclusion: </strong>These pathways taken together help in understanding the overall health beneficial effects of I3C.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"237-247"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880055/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143190120","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":"Pharmacokinetics of Nivolumab and Erythropoietin in a Rat Model of Diet-Induced Obesity.","authors":"Yi-Hua Sheng, Celine Park, Yae Eun Chong, Christine Yohn, Anna Siemiątkowska, Katarzyna Kosicka-Noworzyń, Amrit Kaur, Karan Sapra, Luigi Brunetti, Leonid Kagan","doi":"10.1007/s11095-025-03819-1","DOIUrl":"10.1007/s11095-025-03819-1","url":null,"abstract":"<p><strong>Purpose: </strong>To investigate how obesity affects the pharmacokinetics of biologics in a rat model.</p><p><strong>Method: </strong>Male Long-Evans rats were fed a high-fat diet from the age of 3 weeks and development of obesity was monitored by measuring body size and composition (fat and lean mass). The animals received nivolumab (1 and 8 mg/kg) or recombinant human erythropoietin (rHuEPO, 1000 IU/kg) by intravenous or subcutaneous injection. Serum samples were collected and analyzed using an enzyme-linked immunosorbent assay (ELISA). Endogenous rat IgG was also measured in the nivolumab study. A standard noncompartmental analysis was performed to calculate pharmacokinetic parameters.</p><p><strong>Results: </strong>When dosed at mg/kg of total body weight approach, no significant differences in pharmacokinetics of nivolumab and rHuEPO between lean and obese cohorts were observed despite significant differences in the body composition. Subcutaneous bioavailability of nivolumab was inversely dependent on the dose level.</p><p><strong>Conclusions: </strong>Pharmacokinetic parameters of two biologics tested in this work were not affected by obesity, and mg/kg dosing approach was necessary to achieve equivalent exposure in serum. The results were different from our previous findings of significant effect of obesity on pharmacokinetics of human IgG in rats. Additional studies with other biologics are urgently needed in preclinical and clinical settings.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"271-280"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880188/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143029251","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":"Prediction of the Extent of Blood-Brain Barrier Transport Using Machine Learning and Integration into the LeiCNS-PK3.0 Model.","authors":"Berfin Gülave, Helle W van den Maagdenberg, Luke van Boven, Gerard J P van Westen, Elizabeth C M de Lange, J G Coen van Hasselt","doi":"10.1007/s11095-025-03828-0","DOIUrl":"10.1007/s11095-025-03828-0","url":null,"abstract":"<p><strong>Introduction: </strong>The unbound brain-to-plasma partition coefficient (K_p,uu,BBB) is an essential parameter for predicting central nervous system (CNS) drug disposition using physiologically-based pharmacokinetic (PBPK) modeling. K_p,uu,BBB values for specific compounds are however often unavailable, and are moreover time consuming to obtain experimentally. The aim of this study was to develop a quantitative structure-property relationship (QSPR) model to predict the K_p,uu,BBB and to demonstrate how QSPR-model predictions can be integrated into a physiologically-based pharmacokinetic model for the CNS.</p><p><strong>Methods: </strong>Rat K_p,uu,BBB values were obtained for 98 compounds from literature or in house historical data. For all compounds, 2D and 3D physico-chemical and structural properties were derived using the Molecular Operating Environment (MOE) software. Multiple machine learning (ML) regression models were compared for prediction of the K_p,uu,BBB, including random forest, support vector machines, K-nearest neighbors, and (sparse-) partial least squares. Finally, we demonstrate how the developed QSPR model predictions can be integrated into a CNS PBPK modeling workflow.</p><p><strong>Results: </strong>Among all ML algorithms, a random forest showed the best predictive performance for K_p,uu,BBB on test data with R² value of 0.61 and 61% of all predictions were within twofold error. The obtained K_p,uu,BBB were successfully integrated into the LeiCNS-PK3.0 CNS PBPK model.</p><p><strong>Conclusions: </strong>The developed random forest QSPR model for K_p,uu,BBB prediction was found to have adequate performance, and can support drug discovery and development of novel investigational drugs targeting the CNS in conjunction with CNS PBPK modeling.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"281-289"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880073/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143391563","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":"Development of an Infant Air-Jet Dry Powder Aerosol Delivery System (iDP-ADS) Including a New Multifunctional Bifurcating Two-Prong Nasal Interface.","authors":"Sarah C Strickler, Dale R Farkas, Mohammad A M Momin, Laura Vargas, Ghali Aladwani, Michael Hindle, Worth Longest","doi":"10.1007/s11095-024-03814-y","DOIUrl":"10.1007/s11095-024-03814-y","url":null,"abstract":"<p><strong>Purpose: </strong>To improve the quality of aerosol delivery to infants, the iDP-ADS was advanced to include dual-prong nose-to-lung aerosol administration with a bifurcating interface, consistently monitor lung pressures and control ventilatory parameters with a pressure monitoring and control (PMC) unit, and implement flexible nasal prongs for use across a range of subject sizes.</p><p><strong>Methods: </strong>Four bifurcating flow pathways were integrated into the iDP-ADS and tested in vitro with a full-term infant nose-throat (NT) model for comparison to the performance of a single-prong interface. After selecting the best-performing flow pathway, flexible prong designs were evaluated in the same model and chosen for additional testing. Realistic pulmonary mechanics (PM) and age-appropriate tidal volumes were used to simulate ventilation with the PMC unit and aerosol delivery in full-term and 34-week gestational age preterm NT models.</p><p><strong>Results: </strong>Three of the four bifurcating flow pathways matched the performance of the single-prong design (tracheal filter delivery of ~55%), and the FP4 design with co-flow was selected. A flexible prong version of FP4 produced similar performance to the rigid version. Measurements from the PMC unit demonstrated that consistent air volumes under safe operating pressures could be delivered with a PEEP between 4-6 cmH₂O. Considering aerosol delivery, PM conditions resulted in ~4% decrease in filter deposition but high lung delivery efficiencies of ~45% and ~34% for the full-term and preterm models, respectively.</p><p><strong>Conclusions: </strong>The best-performing interface with flexible prongs matched the lung delivery efficiency of a high-transmission single-prong interface and delivered high aerosol doses through late-preterm to full-term NT models.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"365-384"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880044/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143391560","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":"In-vitro Evaluation of Solution Pressurised Metered Dose Inhaler Sprays with Low-GWP Propellants.","authors":"Daniel J Duke, Lingzhe Rao, Benjamin Myatt, Phil Cocks, Stephen Stein, Nirmal Marasini, Hui Xin Ong, Paul Young","doi":"10.1007/s11095-025-03830-6","DOIUrl":"10.1007/s11095-025-03830-6","url":null,"abstract":"<p><strong>Purpose: </strong>The transition from high Global Warming Potential (GWP) propellants such as HFA134a to low-GWP alternatives such as HFA152a and HFO1234ze(E) in pressurised metered dose inhalers (pMDIs) poses a number of challenges for inhaled pharmaceutical product development. Changes in chemicophysical properties will alter product performance, impacting in-vitro bioequivalence metrics. This study investigates those differences using equivalent pMDI hardware and formulations.</p><p><strong>Methods: </strong>Aerodynamic particle size distribution (APSD) measurements, laser diffraction and high-speed imaging were used to compare the performance of HFA134a, HFA152a and HFO1234ze(E) solution formulations of beclomethasone dipropionate. Propellant-only placebos, cosolvent-free solutions, and ethanol solutions at 8% and 15% w/w were investigated.</p><p><strong>Results: </strong>HFA152a formulations had increased drug deposition on the actuator and throat while HFO1234ze(E) produced comparable APSD performance to HFA134a formulations. Plumes from HFA152a formulations spread more rapidly and were less stable and repeatable than those from HFA134a. HFO1234ze(E) plumes spread more slowly than HFA134a, but converged with HFA134a ex-mouthpiece. Differences between propellants were moderated by the addition of ethanol.</p><p><strong>Conclusion: </strong>Plume stability is a driver of differences between formulations in the near-orifice region. Shot-to-shot repeatability differences are more pronounced ex-mouthpiece, where mixing with ambient air is dominant. Modifications to low-GWP pMDI product actuator orifice and mouthpiece geometries may provide a route to improved in-vitro product bioequivalence relative to current pMDIs. Differences between formulations are modest and may be managed through a combination of formulation, orifice and mouthpiece geometry changes. These generic formulations provide a database of benchmark data against which the performance of low-GWP products may be compared.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"385-400"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880099/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143409564","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":"Nanobody-Oligonucleotide Conjugates (NucleoBodies): The Next Frontier in Oligonucleotide Therapy.","authors":"Rajan K Tripathy, Abhay H Pande","doi":"10.1007/s11095-025-03829-z","DOIUrl":"10.1007/s11095-025-03829-z","url":null,"abstract":"<p><p>As of now, more than 15 oligonucleotide drugs, primarily small interfering RNAs and antisense oligonucleotide classes, have been approved by the US FDA for therapeutic use, and many more are under clinical trials. However, safe and effective delivery of the oligonucleotide-based drugs to the target tissue still remains a major challenge. For enhanced plasma half-life, effective endosomal release, and other multiple functionalities, various carrier molecules have been used over the years. The successful therapeutic application of antibody-drug conjugates has made antibodies a popular choice for the delivery of oligonucleotide payloads into the target tissues. Single-chain variable domains of heavy chain antibodies (nanobodies) have proven a promising alternative to antibodies in recent years due to their small size, high affinity for the target, cell-penetrating potency, simple and easy production. The present review highlights the oligonucleotide drug types and their conjugation with nanobodies called NucleoBodies for effective targeted delivery, detection and diagnostics.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"219-236"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143426034","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 vitro Stability Study of a Panel of Commercial Antibodies at Physiological pH and Temperature as a Guide to Screen Biologic Candidate Molecules for the Potential Risk of In vivo Asparagine Deamidation and Activity Loss.","authors":"Richa Garg, Sean McCarthy, Alayna George Thompson, Jiang Zhang, Emily Mattson, Anca Clabbers, Aimalohi Acquah, Jianwen Xu, Chen Zhou, Amr Ali, Dana Filoti, Rajeeva Singh","doi":"10.1007/s11095-025-03825-3","DOIUrl":"10.1007/s11095-025-03825-3","url":null,"abstract":"<p><strong>Objective: </strong>Biologic drug molecules such as antibodies are exposed to the physiological stress conditions of pH 7.4 and 37°C during their long circulation lifetime in vivo. The stress on biologic molecules in vivo is more severe compared to that under typical storage conditions of low pH formulation and cold temperature. Chemical degradation of critical residues such as asparagine may occur in vivo, leading to potential loss of biological activity. This study describes a physiologically relevant and convenient in vitro PBS stress condition of pH 7.4 and 40°C for pre-clinical stability screening of biologic molecules.</p><p><strong>Methods: </strong>As benchmarks, multiple commercial antibodies (alirocumab, evolocumab, golimumab, ramucirumab, and trastuzumab) were tested in parallel for formulation stability at storage and accelerated temperature conditions and for physiological stability at pH 7.4 and 40°C stress both for 3-4 weeks. The stressed antibodies were monitored for chemical modification and target binding, without requiring affinity purification.</p><p><strong>Results: </strong>The major CDR chemical modifications observed in PBS-stressed commercial antibodies were deamidations of asparagine residues. Although slight decreases in target binding were observed for two antibodies, the affinities overall remained strong after PBS stress.</p><p><strong>Conclusions: </strong>This benchmarking study of commercial antibodies would be useful as a guide to screen discovery-stage biologic molecules both for drug product stability at formulation pH under storage and accelerated temperature conditions and for physiological stability under in vivo-mimicking pH and temperature stress condition.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"353-363"},"PeriodicalIF":3.5,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880144/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468720","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}