Pharmaceutics最新文献

{"title":"Multidrug Combinations against SARS-CoV-2 Using GS-441524 or Ivermectin with Molnupiravir and/or Nirmatrelvir in Reconstituted Human Nasal Airway Epithelia.","authors":"Denise Siegrist, Hulda R Jonsdottir, Mendy Bouveret, Bernadett Boda, Samuel Constant, Olivier B Engler","doi":"10.3390/pharmaceutics16101262","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101262","url":null,"abstract":"<p><p><b>Background.</b> The emergence, global spread, and persistence of SARS-CoV-2 resulted in an unprecedented need for effective antiviral drugs. Throughout the pandemic, various drug development and treatment strategies were adopted, including repurposing of antivirals designed for other viruses along with a multitude of other drugs with varying mechanisms of action (MoAs). Furthermore, multidrug treatment against COVID-19 is an ongoing topic and merits further investigation. <b>Method/Objectives.</b> We assessed the efficacy of multidrug treatment against SARS-CoV-2 in reconstituted human nasal epithelia, using combinations of molnupiravir and nirmatrelvir as a baseline, adding suboptimal concentrations of either GS-441524 or ivermectin, attempting to increase overall antiviral activity while lowering the overall therapeutic dose. <b>Results.</b> Nirmatrelvir combined with molnupiravir, GS-441524, or ivermectin at suboptimal concentrations show increased antiviral activity compared to single treatment. No triple combinations showed improved inhibition of SARS-CoV-2 replication beyond what was observed for double treatments. <b>Conclusions.</b> In general, we observed that the addition of a third compound is not beneficial for antiviral activity, while various double combinations exhibit increased antiviral activity over single treatment.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510096/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505572","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":"Role of In Vitro Tests in the Characterisation of Locally Applied, Locally Acting Drugs in the Throat: Application to Flurbiprofen.","authors":"Vit Perlik, Hafsa Ali, Jean M Cardot, Anuradha Kulasekaran","doi":"10.3390/pharmaceutics16101261","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101261","url":null,"abstract":"<p><p><b>Background/Objectives:</b> For locally applied, locally acting generic drug products, comparison to an originator product based on systemic exposure is usually not feasible due to low plasma concentrations and inadequate reflection of local exposure at the site of action. Where a validated PD model exists, a comparative clinical study can be performed in healthy subjects; where no surrogate endpoint is available, patients with the relevant indication need to be enrolled, with all the associated factors which could result in lack of sensitivity. Even though the need for alternative in vitro approaches has been acknowledged by both industry and regulatory bodies, the complexity of in vivo drug delivery processes makes the development of guidance documents particularly difficult. Our objective was to present in vitro approaches less classically used and to address in vivo relevance of the selected tests. <b>Methods:</b> This article analyses current regulatory approaches in Europe and the U.S., and highlights the key advantages of in vitro tests in terms of their sensitivity, reliability, reproducibility and in vivo relevance using locally applied flurbiprofen in various formulations. <b>Results:</b> The in vitro esophageal retention (IVOR) model demonstrates that the first 6-10 min after application of different flurbiprofen formulations is important for their comparison and also offers the best correlation with in vivo data using the partial area under the concentration-time curves (pAUCs). Rheological evaluations further demonstrated that the mucoadhesive properties of the gel spray formulation are based on interaction with mucin. <b>Conclusions:</b> Designing a relevant in vitro test requires adequate evaluation of the complexity of the drug substance, drug product, dosing conditions and delivery processes.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510169/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505608","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":"Developing Robust Human Liver Microsomal Stability Prediction Models: Leveraging Inter-Species Correlation with Rat Data.","authors":"Pranav Shah, Vishal B Siramshetty, Ewy Mathé, Xin Xu","doi":"10.3390/pharmaceutics16101257","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101257","url":null,"abstract":"<p><p><b>Objectives:</b> Pharmacokinetic issues were the leading cause of drug attrition, accounting for approximately 40% of all cases before the turn of the century. To this end, several high-throughput in vitro assays like microsomal stability have been developed to evaluate the pharmacokinetic profiles of compounds in the early stages of drug discovery. At NCATS, a single-point rat liver microsomal (RLM) stability assay is used as a Tier I assay, while human liver microsomal (HLM) stability is employed as a Tier II assay. We experimentally screened and collected data on over 30,000 compounds for RLM stability and over 7000 compounds for HLM stability. Although HLM stability screening provides valuable insights, the increasing number of hits generated, along with the time- and resource-intensive nature of the assay, highlights the need for alternative strategies. One promising approach is leveraging in silico models trained on these experimental datasets. <b>Methods:</b> We describe the development of an HLM stability prediction model using our in-house HLM stability dataset. <b>Results:</b> Employing both classical machine learning methods and advanced techniques, such as neural networks, we achieved model accuracies exceeding 80%. Moreover, we validated our model using external test sets and found that our models are comparable to some of the best models in literature. Additionally, the strong correlation observed between our RLM and HLM data was further reinforced by the fact that our HLM model performance improved when using RLM stability predictions as an input descriptor. <b>Conclusions:</b> The best model along with a subset of our dataset (PubChem AID: 1963597) has been made publicly accessible on the ADME@NCATS website for the benefit of the greater drug discovery community. To the best of our knowledge, it is the largest open-source model of its kind and the first to leverage cross-species data.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510424/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505634","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":"Quality by Design (QbD) Approach to Develop Colon-Specific Ketoprofen Hot-Melt Extruded Pellets: Impact of Eudragit^® S 100 Coating on the In Vitro Drug Release.","authors":"Sateesh Kumar Vemula, Sagar Narala, Prateek Uttreja, Nagarjuna Narala, Bhaskar Daravath, Chamundeswara Srinivasa Akash Kalla, Srikanth Baisa, Siva Ram Munnangi, Naveen Chella, Michael A Repka","doi":"10.3390/pharmaceutics16101265","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101265","url":null,"abstract":"<p><strong>Background: </strong>A pelletizer paired with hot-melt extrusion technology (HME) was used to develop colon-targeted pellets for ketoprofen (KTP). Thermal stability and side effects in the upper gastrointestinal tract made ketoprofen more suitable for this work.</p><p><strong>Methods: </strong>The pellets were prepared using the enzyme-triggered polymer Pectin LM in the presence of HPMC HME 4M, followed by pH-dependent Eudragit^® S 100 coating to accommodate the maximum drug release in the colon by minimizing drug release in the upper gastrointestinal tract (GIT). Box-Behnken Design (BBD) was used for response surface optimization of the proportion of different independent variables like Pectin LM (A), HPMC HME 4M (B), and Eudragit^® S 100 (C) required to lower the early drug release in upper GIT and to extend the drug release in the colon.</p><p><strong>Results: </strong>Solid-state characterization studies revealed that ketoprofen was present in a solid solution state in the hot-melt extruded polymer matrix. The desired responses of the prepared optimized KTP pellets obtained by considering the designed space showed 1.20% drug release in 2 h, 3.73% in the first 5 h of the lag period with the help of Eudragit^® S 100 coating, and 93.96% in extended release up to 24 h in the colonic region.</p><p><strong>Conclusions: </strong>Hence, developing Eudragit-coated hot-melt extruded pellets could be a significant method for achieving the colon-specific release of ketoprofen.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11509973/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505595","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 Halloysite Nanohybrids-Based Films: Enhancing Mechanical and Hydrophilic Properties for Wound Healing.","authors":"Francisco Ramón Rodríguez Pozo, Daiana Ianev, Tomás Martínez Rodríguez, José L Arias, Fátima Linares, Carlos Miguel Gutiérrez Ariza, Caterina Valentino, Francisco Arrebola Vargas, Pablo Hernández Benavides, José Manuel Paredes, María Del Mar Medina Pérez, Silvia Rossi, Giuseppina Sandri, Carola Aguzzi","doi":"10.3390/pharmaceutics16101258","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101258","url":null,"abstract":"<p><p>Most of the therapeutic systems developed for managing chronic skin wounds lack adequate mechanical and hydration properties, primarily because they rely on a single component. This study addresses this issue by combining organic and inorganic materials to obtain hybrid films with enhanced mechanical behavior, adhesion, and fluid absorption properties. To that aim, chitosan/hydrolyzed collagen blends were mixed with halloysite/antimicrobial nanohybrids at 10% and 20% (<i>w</i>/<i>w</i>) using glycerin or glycerin/polyethylene glycol-1500 as plasticizers. The films were characterized through the use of Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and electron microscopy. The mechanical properties were evaluated macroscopically using tensile tests, and at a nanoscale through atomic force microscopy (AFM) and nanoindentation. Thermodynamic studies were conducted to assess their hydrophilic or hydrophobic character. Additionally, in vitro cytocompatibility tests were performed on human keratinocytes. Results from FTIR, TGA, AFM and electron microscopy confirmed the hybrid nature of the films. Both tensile tests and nanomechanical measurements postulated that the nanohybrids improved the films' toughness and adhesion and optimized the nanoindentation properties. All nanohybrid-loaded films were hydrophilic and non-cytotoxic, showcasing their potential for skin wound applications given their enhanced performance at the macro- and nanoscale.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11509966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505641","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":"Electrospun PCL Nerve Wrap Coated with Graphene Oxide Supports Axonal Growth in a Rat Sciatic Nerve Injury Model.","authors":"Meaghan E Harley-Troxell, Richard Steiner, Steven D Newby, Austin J Bow, Thomas J Masi, Nicholas Millis, Alicia Adina Matavosian, Dustin Crouch, Stacy Stephenson, David E Anderson, Madhu Dhar","doi":"10.3390/pharmaceutics16101254","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101254","url":null,"abstract":"<p><p><b>Background/Objectives:</b> Peripheral nerve injuries (PNIs) are a debilitating problem, resulting in diminished quality of life due to the continued presence of both chronic and acute pain. The current standard of practice for the repair of PNIs larger than 10 mm is the use of autologous nerve grafts. Autologous nerve grafts have limitations that often result in outcomes that are not sufficient to remove motor and sensory impairments. Bio-mimetic nanocomposite scaffolds combined with mesenchymal stem cells (MSCs) represent a promising approach for PNIs. In this study, we investigated the potential of an electrospun wrap of polycaprolactone (PCL) + graphene oxide (GO), with and without xenogeneic human adipose tissue-derived MSCs (hADMSCs) to use as a platform for neural tissue engineering. <b>Methods:</b> We evaluated, in vitro and in vivo, the potential of the nerve wrap in providing support for axonal growth. To establish the rat sciatic nerve defect model, a 10 mm long limiting defect was created in the rat sciatic nerve of 18 Lewis rats. Rats treated with the nanocomposites were compared with autograft-treated defects. Gait, histological, and muscle analyses were performed after sacrifice at 12 weeks post-surgery. <b>Results:</b> Our findings demonstrate that hADMSCs had the potential to transdifferentiate into neural lineage and that the nanocomposite successfully delivered hADMSCs to the injury site. Histologically, we show that the PCL + GO nanocomposite with hADMSCs is comparable to the autologous nerve graft, to support and guide axonal growth. <b>Conclusions:</b> The novel PCL + GO nerve wrap and hADMSCs used in this study provide a foundation on which to build upon and generate future strategies for PNI repair.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510652/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505646","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":"Experimental and Modelling Study of Controlled Release from Dextran-Based Cryogels.","authors":"Carolina Lauriola, Laura Di Muzio, Patrizia Paolicelli, Maria Antonietta Casadei, Claudia Sergi, Jacopo Tirillò, Vito Cosimo Carriero, Alessandra Adrover","doi":"10.3390/pharmaceutics16101256","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101256","url":null,"abstract":"<p><p>In this work, five different dextran-based cryogels for controlled drug release are investigated. Vitamin B12 was used as a model drug for in vitro release tests. Two different drug-loading procedures were adopted, leading to very different drug release curves. Indeed, a fast Fickian release was observed when freeze-dried samples of DEX₄₀PEG₃₆₀MA and DEX₄₀PEG₅₀₀MA were infused with the drug after cryogel formation. On the contrary, a slowed highly non-Fickian behavior arises when the drug is loaded before the low-temperature crosslinking step, leading to the cryogel formation. The non-Fickian drug release, observed for all the five different dextran-based cryogels investigated, is actually due to the cryoconcentration phenomenon, modeled with a two-step release process. The proposed transport model accurately predicts experimental release curves characterized by a long lag time, confirming that dextran-based cryogels are suitable for controlled release.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510673/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505653","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":"Tailoring Crystallization Kinetics in Thin Sucrose Films during Convective Drying: Impact of Temperature and Humidity on Onset, Growth, and Nucleation Rate.","authors":"Martin Schugmann, Petra Foerst","doi":"10.3390/pharmaceutics16101260","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101260","url":null,"abstract":"<p><p>Drying experiments with varying air temperature and humidity were conducted to investigate the influence of the drying process on the crystallization of thin sucrose films. For the first time, the effects of the nucleation onset, nucleation rate, and growth rate were investigated in situ and their differentiated influence on product crystallinity could be assessed. The growth rate was not influenced by air humidity but showed a strong dependence on temperature. It increased with drying temperature; however, at high temperatures, growth was inhibited when the water content falls below a critical level. Noticeable differences in nucleation behavior could be observed with regard to air humidity. Dry air led to crystallization onsets at lower levels of supersaturation, while moderately humid air retarded it. At higher temperatures, nucleation onset commenced at lower water contents but at a constant supersaturation level. The nucleation rate doubled in experiments with moderately humid air (15% RH), while an elevated drying temperature showed generally lower nucleation rates. The observed differences in the nucleation onset and rate could be explained by the film-internal concentration profile, which is strongly influenced by drying parameters. The insights therefore provide a differentiated understanding of the formation of the physical state and how it can be influenced during convective drying.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510720/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142516565","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":"Self-Tumor Antigens in Solid Tumors Turned into Vaccines by α-gal Micelle Immunotherapy.","authors":"Uri Galili","doi":"10.3390/pharmaceutics16101263","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101263","url":null,"abstract":"<p><p>A major reason for the failure of the immune system to detect tumor antigens (TAs) is the insufficient uptake, processing, and presentation of TAs by antigen-presenting cells (APCs). The immunogenicity of TAs in the individual patient can be markedly increased by the in situ targeting of tumor cells for robust uptake by APCs, without the need to identify and characterize the TAs. This is feasible by the intra-tumoral injection of α-gal micelles comprised of glycolipids presenting the carbohydrate-antigen \"α-gal epitope\" (Galα1-3Galβ1-4GlcNAc-R). Humans produce a natural antibody called \"anti-Gal\" (constituting ~1% of immunoglobulins), which binds to α-gal epitopes. Tumor-injected α-gal micelles spontaneously insert into tumor cell membranes, so that multiple α-gal epitopes are presented on tumor cells. Anti-Gal binding to these epitopes activates the complement system, resulting in the killing of tumor cells, and the recruitment of multiple APCs (dendritic cells and macrophages) into treated tumors by the chemotactic complement cleavage peptides C5a and C3a. In this process of converting the treated tumor into a personalized TA vaccine, the recruited APC phagocytose anti-Gal opsonized tumor cells and cell membranes, process the internalized TAs and transport them to regional lymph-nodes. TA peptides presented on APCs activate TA-specific T cells to proliferate and destroy the metastatic tumor cells presenting the TAs. Studies in anti-Gal-producing mice demonstrated the induction of effective protection against distant metastases of the highly tumorigenic B16 melanoma following injection of natural and synthetic α-gal micelles into primary tumors. This treatment was further found to synergize with checkpoint inhibitor therapy by the anti-PD1 antibody. Phase-1 clinical trials indicated that α-gal micelle immunotherapy is safe and can induce the infiltration of CD4+ and CD8+ T cells into untreated distant metastases. It is suggested that, in addition to converting treated metastases into an autologous TA vaccine, this treatment should be considered as a neoadjuvant therapy, administering α-gal micelles into primary tumors immediately following their detection. Such an immunotherapy will convert tumors into a personalized anti-TA vaccine for the period prior to their resection.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510312/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505610","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":"Novel Anti-Trop2 Nanobodies Disrupt Receptor Dimerization and Inhibit Tumor Cell Growth.","authors":"Junwen Deng, Zhongmin Geng, Linli Luan, Dingwen Jiang, Jian Lu, Hanzhong Zhang, Bingguan Chen, Xinlin Liu, Dongming Xing","doi":"10.3390/pharmaceutics16101255","DOIUrl":"https://doi.org/10.3390/pharmaceutics16101255","url":null,"abstract":"<p><p><b>Background:</b> Trop2 (trophoblast cell-surface antigen 2) is overexpressed in multiple malignancies and is closely associated with poor prognosis, thus positioning it as a promising target for pan-cancer therapies. Despite the approval of Trop2-targeted antibody-drug conjugates (ADCs), challenges such as side effects, drug resistance, and limited efficacy persist. Recent studies have shown that the dimeric forms of Trop2 are crucial for its oncogenic functions, and the binding epitopes of existing Trop2-targeted drugs lie distant from the dimerization interface, potentially limiting their antitumor efficacy. <b>Method:</b> A well-established synthetic nanobody library was screened against Trop2-ECD. The identified nanobodies were extensively characterized, including their binding specificity and affinity, as well as their bioactivities in antigen-antibody endocytosis, cell proliferation, and the inhibition of Trop2 dimer assembly. Finally, ELISA based epitope analysis and AlphaFold 3 were employed to elucidate the binding modes of the nanobodies. <b>Results:</b> We identified two nanobodies, N14 and N152, which demonstrated high affinity and specificity for Trop2. Cell-based assays confirmed that N14 and N152 can facilitate receptor internalization and inhibit growth in Trop2-positive tumor cells. Epitope analysis uncovered that N14 and N152 are capable of binding with all three subdomains of Trop2-ECD and effectively disrupt Trop2 dimerization. Predictive modeling suggests that N14 and N152 likely target the epitopes at the interface of Trop2 <i>cis</i>-dimerization. The binding modality and mechanism of action demonstrated by N14 and N152 are unique among Trop2-targeted antibodies. <b>Conclusions:</b> we identified two novel nanobodies, N14 and N152, that specifically bind to Trop2. Importantly, these nanobodies exhibit significant anti-tumor efficacy and distinctive binding patterns, underscoring their potential as innovative Trop2-targeted therapeutics.</p>","PeriodicalId":19894,"journal":{"name":"Pharmaceutics","volume":"16 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510716/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505574","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}