Pub Date : 2024-09-30DOI: 10.1021/acs.molpharmaceut.4c00758
Tycho Heimbach, Flora Musuamba Tshinanu, Kimberly Raines, Luiza Borges, Shinichi Kijima, Maria Malamatari, Rebecca Moody, Shereeni Veerasingham, Paul Seo, David Turner, Lanyan Fang, Cordula Stillhart, Philip Bransford, Xiaojun Ren, Nikunjkumar Patel, David Sperry, Hansong Chen, Amin Rostami-Hodjegan, Viera Lukacova, Duxin Sun, Jean-Flaubert Nguefack, Tessa Carducci, Manuela Grimstein, Xavier Pepin, Masoud Jamei, Konstantinos Stamatopoulos, Min Li, Maitri Sanghavi, Christer Tannergren, Haritha Mandula, Zhuojun Zhao, Tzuchi Rob Ju, Christian Wagner, Sumit Arora, Michael Wang, Gregory Rullo, Amitava Mitra, Sivacharan Kollipara, Siri Kalyan Chirumamilla, James E Polli, Claire Mackie
The proceedings from the 30th August 2023 (Day 2) of the workshop "Physiologically Based Biopharmaceutics Models (PBBM) Best Practices for Drug Product Quality: Regulatory and Industry Perspectives" are provided herein. Day 2 covered PBBM case studies from six regulatory authorities which provided considerations for model verification, validation, and application based on the context of use (COU) of the model. PBBM case studies to define critical material attribute (CMA) specification settings, such as active pharmaceutical ingredient (API) particle size distributions (PSDs) were shared. PBBM case studies to define critical quality attributes (CQAs) such as the dissolution specification setting or to define the bioequivalence safe space were also discussed. Examples of PBBM using the credibility assessment framework, COU and model risk assessment, as well as scientific learnings from PBBM case studies are provided. Breakout session discussions highlighted current trends and barriers to application of PBBMs including: (a) PBBM credibility assessment framework and level of validation, (b) use of disposition parameters in PBBM and points to consider when iv data are not available, (c) conducting virtual bioequivalence trials and dealing with variability, (d) model acceptance criteria, and (e) application of PBBMs for establishing safe space and failure edges.
{"title":"PBBM Considerations for Base Models, Model Validation, and Application Steps: Workshop Summary Report.","authors":"Tycho Heimbach, Flora Musuamba Tshinanu, Kimberly Raines, Luiza Borges, Shinichi Kijima, Maria Malamatari, Rebecca Moody, Shereeni Veerasingham, Paul Seo, David Turner, Lanyan Fang, Cordula Stillhart, Philip Bransford, Xiaojun Ren, Nikunjkumar Patel, David Sperry, Hansong Chen, Amin Rostami-Hodjegan, Viera Lukacova, Duxin Sun, Jean-Flaubert Nguefack, Tessa Carducci, Manuela Grimstein, Xavier Pepin, Masoud Jamei, Konstantinos Stamatopoulos, Min Li, Maitri Sanghavi, Christer Tannergren, Haritha Mandula, Zhuojun Zhao, Tzuchi Rob Ju, Christian Wagner, Sumit Arora, Michael Wang, Gregory Rullo, Amitava Mitra, Sivacharan Kollipara, Siri Kalyan Chirumamilla, James E Polli, Claire Mackie","doi":"10.1021/acs.molpharmaceut.4c00758","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00758","url":null,"abstract":"<p><p>The proceedings from the 30th August 2023 (Day 2) of the workshop \"Physiologically Based Biopharmaceutics Models (PBBM) Best Practices for Drug Product Quality: Regulatory and Industry Perspectives\" are provided herein. Day 2 covered PBBM case studies from six regulatory authorities which provided considerations for model verification, validation, and application based on the context of use (COU) of the model. PBBM case studies to define critical material attribute (CMA) specification settings, such as active pharmaceutical ingredient (API) particle size distributions (PSDs) were shared. PBBM case studies to define critical quality attributes (CQAs) such as the dissolution specification setting or to define the bioequivalence safe space were also discussed. Examples of PBBM using the credibility assessment framework, COU and model risk assessment, as well as scientific learnings from PBBM case studies are provided. Breakout session discussions highlighted current trends and barriers to application of PBBMs including: (a) PBBM credibility assessment framework and level of validation, (b) use of disposition parameters in PBBM and points to consider when iv data are not available, (c) conducting virtual bioequivalence trials and dealing with variability, (d) model acceptance criteria, and (e) application of PBBMs for establishing safe space and failure edges.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337280","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-30DOI: 10.1021/acs.molpharmaceut.4c00859
Yuhan Wang, Hywel D Williams, Duygu Dikicioglu, Paul A Dalby
Computational methods including machine learning and molecular dynamics simulations have strong potential to characterize, understand, and ultimately predict the properties of proteins relevant to their stability and function as therapeutics. Such methods would streamline the development pathway by minimizing the current experimental testing required for many protein variants and formulations. The molecular understanding of thermostability and aggregation propensity has advanced significantly along with predictive algorithms based on the sequence-level or structural-level information on a protein. However, these approaches focus largely on a comparison of protein sequence variations to correlate the properties of proteins to their stability, solubility, and aggregation propensity. For therapeutic protein development, it is of equal importance to take into account the impact of the formulation conditions to elucidate and predict the stability of the antibody drugs. At the macroscopic level, changing temperature, pH, ionic strength, and the addition of excipients can significantly alter the kinetics of protein aggregation. The mechanisms controlling aggregation kinetics have been traced back to a combination of molecular features, including conformational stability, partial unfolding to aggregation-prone states, and the colloidal stability governed by surface charges and hydrophobicity. However, very little has been done to evaluate these features in the context of protein dynamics in different formulations. In this work, we have combined a range of molecular features calculated from the Fab A33 protein sequence and molecular dynamics simulations. Using the power of advanced, yet interpretable, statistical tools, it has been possible to uncover greater insights into the mechanisms behind protein stability, validating previous findings, and also develop models that can predict the aggregation kinetics within a range of 49 different solution conditions.
包括机器学习和分子动力学模拟在内的计算方法在表征、理解并最终预测与蛋白质稳定性和治疗功能相关的蛋白质特性方面具有强大的潜力。这些方法将最大限度地减少目前对许多蛋白质变体和制剂所需的实验测试,从而简化开发途径。随着基于蛋白质序列级或结构级信息的预测算法的发展,对热稳定性和聚集倾向的分子理解也有了长足的进步。不过,这些方法主要侧重于比较蛋白质的序列变化,从而将蛋白质的特性与其稳定性、可溶性和聚集倾向联系起来。对于治疗性蛋白质开发而言,考虑制剂条件的影响以阐明和预测抗体药物的稳定性同样重要。在宏观层面上,改变温度、pH 值、离子强度和添加辅料会显著改变蛋白质的聚集动力学。控制聚集动力学的机制可追溯到分子特征的组合,包括构象稳定性、部分展开到易聚集状态,以及由表面电荷和疏水性决定的胶体稳定性。然而,在不同制剂的蛋白质动力学背景下对这些特征进行评估的工作却少之又少。在这项研究中,我们结合了从 Fab A33 蛋白序列和分子动力学模拟中计算出的一系列分子特征。利用先进但可解释的统计工具,我们得以更深入地了解蛋白质稳定性背后的机制,验证了之前的研究结果,同时还开发了可预测 49 种不同溶液条件下聚集动力学的模型。
{"title":"Predictive Model Building for Aggregation Kinetics Based on Molecular Dynamics Simulations of an Antibody Fragment.","authors":"Yuhan Wang, Hywel D Williams, Duygu Dikicioglu, Paul A Dalby","doi":"10.1021/acs.molpharmaceut.4c00859","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00859","url":null,"abstract":"<p><p>Computational methods including machine learning and molecular dynamics simulations have strong potential to characterize, understand, and ultimately predict the properties of proteins relevant to their stability and function as therapeutics. Such methods would streamline the development pathway by minimizing the current experimental testing required for many protein variants and formulations. The molecular understanding of thermostability and aggregation propensity has advanced significantly along with predictive algorithms based on the sequence-level or structural-level information on a protein. However, these approaches focus largely on a comparison of protein sequence variations to correlate the properties of proteins to their stability, solubility, and aggregation propensity. For therapeutic protein development, it is of equal importance to take into account the impact of the formulation conditions to elucidate and predict the stability of the antibody drugs. At the macroscopic level, changing temperature, pH, ionic strength, and the addition of excipients can significantly alter the kinetics of protein aggregation. The mechanisms controlling aggregation kinetics have been traced back to a combination of molecular features, including conformational stability, partial unfolding to aggregation-prone states, and the colloidal stability governed by surface charges and hydrophobicity. However, very little has been done to evaluate these features in the context of protein dynamics in different formulations. In this work, we have combined a range of molecular features calculated from the Fab A33 protein sequence and molecular dynamics simulations. Using the power of advanced, yet interpretable, statistical tools, it has been possible to uncover greater insights into the mechanisms behind protein stability, validating previous findings, and also develop models that can predict the aggregation kinetics within a range of 49 different solution conditions.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-27DOI: 10.1021/acs.molpharmaceut.4c00751
Da Hye Yang, Saeed Najafian, Bodhisattwa Chaudhuri, Na Li
The particle drifting effect, where nanosized colloidal drug particles overcome the diffusional resistance of the aqueous boundary layer adjacent to the intestinal wall and increase drug absorption rates, is drawing increasing attention in pharmaceutical research. However, mechanistic understanding and accurate prediction of the particle drifting effect remain lacking. In this study, we systematically evaluated the extent of the particle drifting effect affected by drug and colloidal properties, including the size, number, and type of the moving species using biphasic diffusion experiments combined with computational fluid dynamics simulations and mass transport analyses. The results showed that the particle drifting effect is a sequential reaction of particle dissolution/dissociation in the diffusional boundary layer, followed by absorption of the free drug. Therefore, factors affecting the rate-limiting step, which can be either process or both under different circumstances, alter the particle drifting effect. Experimental results also agree with the theory that the particle dissolution rate is dependent on particle size, concentration, and drug solubility. In addition, rapid bile micelle dissociation and bile salt absorption facilitated drug absorption by the particle drifting effect. Our findings explain the highly dynamic nature of the particle drifting effect and will contribute to rational formulation development and better bioavailability prediction for formulations containing colloidal particles.
{"title":"The Particle Drifting Effect: A Combined Function of Colloidal and Drug Properties.","authors":"Da Hye Yang, Saeed Najafian, Bodhisattwa Chaudhuri, Na Li","doi":"10.1021/acs.molpharmaceut.4c00751","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00751","url":null,"abstract":"<p><p>The particle drifting effect, where nanosized colloidal drug particles overcome the diffusional resistance of the aqueous boundary layer adjacent to the intestinal wall and increase drug absorption rates, is drawing increasing attention in pharmaceutical research. However, mechanistic understanding and accurate prediction of the particle drifting effect remain lacking. In this study, we systematically evaluated the extent of the particle drifting effect affected by drug and colloidal properties, including the size, number, and type of the moving species using biphasic diffusion experiments combined with computational fluid dynamics simulations and mass transport analyses. The results showed that the particle drifting effect is a sequential reaction of particle dissolution/dissociation in the diffusional boundary layer, followed by absorption of the free drug. Therefore, factors affecting the rate-limiting step, which can be either process or both under different circumstances, alter the particle drifting effect. Experimental results also agree with the theory that the particle dissolution rate is dependent on particle size, concentration, and drug solubility. In addition, rapid bile micelle dissociation and bile salt absorption facilitated drug absorption by the particle drifting effect. Our findings explain the highly dynamic nature of the particle drifting effect and will contribute to rational formulation development and better bioavailability prediction for formulations containing colloidal particles.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lipid nanoparticle-encapsulated mRNA (mRNA-LNP) vaccines have been approved for use to combat coronavirus disease 2019 (COVID-19). The mRNA-LNPs contain PEG-conjugated lipids. Clinical studies have reported that mRNA-LNPs induce the production of anti-PEG antibodies, but the anti-PEG antibodies do not affect the production of neutralizing antibodies. However, the detailed influence of anti-PEG antibodies on mRNA-LNP vaccines remains unclear. Therefore, in this study, we prepared ovalbumin (OVA) as a model antigen-encoding mRNA-loaded LNP (mRNA-OVA-LNP), and we determined whether anti-PEG antibodies could affect the antigen-specific immune response of mRNA-OVA-LNP vaccination in mice pretreated with PEG-modified liposomes to induce the production of anti-PEG antibodies. After intramuscular (i.m.) injection of the mRNA-LNP, the anti-PEG antibodies did not change the expression of protein or induction of cytokine and cellular immune response but did slightly increase the induction of antigen-specific antibodies. Furthermore, repeated mRNA-LNP i.m. injection induced the production of anti-PEG IgM and anti-PEG IgG. Our results suggest that mRNA-LNP induces the production of anti-PEG antibodies, but the priming of the antigen-specific immune response of mRNA-LNP vaccination is not notably affected by anti-PEG antibodies.
{"title":"Effect of Anti-PEG Antibody on Immune Response of mRNA-Loaded Lipid Nanoparticles.","authors":"Daiki Omata, Eigo Kawahara, Lisa Munakata, Hiroki Tanaka, Hidetaka Akita, Yasuo Yoshioka, Ryo Suzuki","doi":"10.1021/acs.molpharmaceut.4c00628","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00628","url":null,"abstract":"<p><p>Lipid nanoparticle-encapsulated mRNA (mRNA-LNP) vaccines have been approved for use to combat coronavirus disease 2019 (COVID-19). The mRNA-LNPs contain PEG-conjugated lipids. Clinical studies have reported that mRNA-LNPs induce the production of anti-PEG antibodies, but the anti-PEG antibodies do not affect the production of neutralizing antibodies. However, the detailed influence of anti-PEG antibodies on mRNA-LNP vaccines remains unclear. Therefore, in this study, we prepared ovalbumin (OVA) as a model antigen-encoding mRNA-loaded LNP (mRNA-OVA-LNP), and we determined whether anti-PEG antibodies could affect the antigen-specific immune response of mRNA-OVA-LNP vaccination in mice pretreated with PEG-modified liposomes to induce the production of anti-PEG antibodies. After intramuscular (i.m.) injection of the mRNA-LNP, the anti-PEG antibodies did not change the expression of protein or induction of cytokine and cellular immune response but did slightly increase the induction of antigen-specific antibodies. Furthermore, repeated mRNA-LNP i.m. injection induced the production of anti-PEG IgM and anti-PEG IgG. Our results suggest that mRNA-LNP induces the production of anti-PEG antibodies, but the priming of the antigen-specific immune response of mRNA-LNP vaccination is not notably affected by anti-PEG antibodies.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Given the aging populations in advanced countries globally, many pharmaceutical companies have focused on developing central nervous system (CNS) drugs. However, due to the blood–brain barrier, drugs do not easily reach the target area in the brain. Although conventional screening methods for drug discovery involve the measurement of (unbound fraction of drug) brain-to-plasma partition coefficients, it is difficult to consider nonequilibrium between plasma and brain compound concentration–time profiles. To truly understand the pharmacokinetics/pharmacodynamics of CNS drugs, compound concentration–time profiles in the brain are necessary; however, such analyses are costly and time-consuming and require a significant number of animals. Therefore, in this study, we attempted to develop an in silico prediction method that does not require a large amount of experimental data by combining modeling and simulation (M&S) with machine learning (ML). First, we constructed a hybrid model linking plasma concentration–time profile to the brain compartment that takes into account the transit time and brain distribution of each compound. Using mouse plasma and brain time experimental values for 103 compounds, we determined the brain kinetic parameters of the hybrid model for each compound; this case was defined as scenario I (a positive control experiment) and included the full brain concentration–time profile data. Next, we built an ML model using chemical structure descriptors as explanatory variables and rate parameters as the target variable, and we then input the predicted values from 5-fold cross-validation (CV) into the hybrid model; this case was defined as scenario II, in which no brain compound concentration–time profile data exist. Finally, for scenario III, assuming that the brain concentration is obtained at only one time point, we used the brain kinetic parameters from the result of the 5-fold CV in scenario II as the initial values for the hybrid model and performed parameter refitting against the observed brain concentration at that time point. As a result, the RMSE/R2-values of the brain compound concentration–time profiles over time were 0.445/0.517 in scenario II and 0.246/0.805 in scenario III, indicating the method provides high accuracy and suggesting that it is a practical method for predicting brain compound concentration–time profiles.
鉴于全球先进国家的人口老龄化问题,许多制药公司都把重点放在开发中枢神经系统(CNS)药物上。然而,由于血脑屏障的存在,药物并不容易到达大脑中的目标区域。虽然药物发现的传统筛选方法涉及测量(药物未结合部分)脑-血浆分配系数,但很难考虑血浆和大脑化合物浓度-时间曲线之间的非平衡。要真正了解中枢神经系统药物的药代动力学/药效学,就需要脑内的化合物浓度-时间曲线;然而,这种分析既费钱又费时,而且需要大量动物。因此,在本研究中,我们尝试通过将建模和仿真(M&S)与机器学习(ML)相结合,开发一种无需大量实验数据的硅学预测方法。首先,我们构建了一个将血浆浓度-时间曲线与脑区联系起来的混合模型,该模型考虑了每种化合物的转运时间和脑区分布。利用 103 种化合物的小鼠血浆和大脑时间实验值,我们确定了混合模型中每种化合物的大脑动力学参数;这种情况被定义为情景 I(阳性对照实验),包括完整的大脑浓度-时间曲线数据。接下来,我们建立了一个以化学结构描述符为解释变量、速率参数为目标变量的 ML 模型,然后将 5 倍交叉验证(CV)的预测值输入混合模型;这种情况被定义为情景 II,即不存在脑部化合物浓度-时间曲线数据。最后,对于情景 III,假设只在一个时间点获得脑部浓度,我们将情景 II 中 5 倍交叉验证结果中的脑部动力学参数作为混合模型的初始值,并根据该时间点的观察脑部浓度进行参数再拟合。结果,脑部化合物浓度-时间曲线随时间变化的 RMSE/R2- 值在方案 II 中为 0.445/0.517,在方案 III 中为 0.246/0.805,表明该方法具有较高的准确性,是预测脑部化合物浓度-时间曲线的实用方法。
{"title":"A Practical In Silico Method for Predicting Compound Brain Concentration–Time Profiles: Combination of PK Modeling and Machine Learning","authors":"Koichi Handa*, Daichi Fujita, Mariko Hirano, Saki Yoshimura, Michiharu Kageyama and Takeshi Iijima, ","doi":"10.1021/acs.molpharmaceut.4c0058410.1021/acs.molpharmaceut.4c00584","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00584https://doi.org/10.1021/acs.molpharmaceut.4c00584","url":null,"abstract":"<p >Given the aging populations in advanced countries globally, many pharmaceutical companies have focused on developing central nervous system (CNS) drugs. However, due to the blood–brain barrier, drugs do not easily reach the target area in the brain. Although conventional screening methods for drug discovery involve the measurement of (unbound fraction of drug) brain-to-plasma partition coefficients, it is difficult to consider nonequilibrium between plasma and brain compound concentration–time profiles. To truly understand the pharmacokinetics/pharmacodynamics of CNS drugs, compound concentration–time profiles in the brain are necessary; however, such analyses are costly and time-consuming and require a significant number of animals. Therefore, in this study, we attempted to develop an <i>in silico</i> prediction method that does not require a large amount of experimental data by combining modeling and simulation (M&S) with machine learning (ML). First, we constructed a hybrid model linking plasma concentration–time profile to the brain compartment that takes into account the transit time and brain distribution of each compound. Using mouse plasma and brain time experimental values for 103 compounds, we determined the brain kinetic parameters of the hybrid model for each compound; this case was defined as scenario I (a positive control experiment) and included the full brain concentration–time profile data. Next, we built an ML model using chemical structure descriptors as explanatory variables and rate parameters as the target variable, and we then input the predicted values from 5-fold cross-validation (CV) into the hybrid model; this case was defined as scenario II, in which no brain compound concentration–time profile data exist. Finally, for scenario III, assuming that the brain concentration is obtained at only one time point, we used the brain kinetic parameters from the result of the 5-fold CV in scenario II as the initial values for the hybrid model and performed parameter refitting against the observed brain concentration at that time point. As a result, the RMSE/R2-values of the brain compound concentration–time profiles over time were 0.445/0.517 in scenario II and 0.246/0.805 in scenario III, indicating the method provides high accuracy and suggesting that it is a practical method for predicting brain compound concentration–time profiles.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142403625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-26DOI: 10.1021/acs.molpharmaceut.4c00604
Courtney Culkins, Roman Adomanis, Nathan Phan, Blaise Robinson, Ethan Slaton, Elijah Lothrop, Yinuo Chen, Blaise R Kimmel
The blood-brain barrier (BBB) is a highly selective network of various cell types that acts as a filter between the blood and the brain parenchyma. Because of this, the BBB remains a major obstacle for drug delivery to the central nervous system (CNS). In recent years, there has been a focus on developing various modifiable platforms, such as monoclonal antibodies (mAbs), nanobodies (Nbs), peptides, and nanoparticles, as both therapeutic agents and carriers for targeted drug delivery to treat brain cancers and diseases. Methods for bypassing the BBB can be invasive or noninvasive. Invasive techniques, such as transient disruption of the BBB using low pulse electrical fields and intracerebroventricular infusion, lack specificity and have numerous safety concerns. In this review, we will focus on noninvasive transport mechanisms that offer high levels of biocompatibility, personalization, specificity and are regarded as generally safer than their invasive counterparts. Modifiable platforms can be designed to noninvasively traverse the BBB through one or more of the following pathways: passive diffusion through a physio-pathologically disrupted BBB, adsorptive-mediated transcytosis, receptor-mediated transcytosis, shuttle-mediated transcytosis, and somatic gene transfer. Through understanding the noninvasive pathways, new applications, including Chimeric Antigen Receptors T-cell (CAR-T) therapy, and approaches for drug delivery across the BBB are emerging.
{"title":"Unlocking the Gates: Therapeutic Agents for Noninvasive Drug Delivery Across the Blood-Brain Barrier.","authors":"Courtney Culkins, Roman Adomanis, Nathan Phan, Blaise Robinson, Ethan Slaton, Elijah Lothrop, Yinuo Chen, Blaise R Kimmel","doi":"10.1021/acs.molpharmaceut.4c00604","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00604","url":null,"abstract":"<p><p>The blood-brain barrier (BBB) is a highly selective network of various cell types that acts as a filter between the blood and the brain parenchyma. Because of this, the BBB remains a major obstacle for drug delivery to the central nervous system (CNS). In recent years, there has been a focus on developing various modifiable platforms, such as monoclonal antibodies (mAbs), nanobodies (Nbs), peptides, and nanoparticles, as both therapeutic agents and carriers for targeted drug delivery to treat brain cancers and diseases. Methods for bypassing the BBB can be invasive or noninvasive. Invasive techniques, such as transient disruption of the BBB using low pulse electrical fields and intracerebroventricular infusion, lack specificity and have numerous safety concerns. In this review, we will focus on noninvasive transport mechanisms that offer high levels of biocompatibility, personalization, specificity and are regarded as generally safer than their invasive counterparts. Modifiable platforms can be designed to noninvasively traverse the BBB through one or more of the following pathways: passive diffusion through a physio-pathologically disrupted BBB, adsorptive-mediated transcytosis, receptor-mediated transcytosis, shuttle-mediated transcytosis, and somatic gene transfer. Through understanding the noninvasive pathways, new applications, including Chimeric Antigen Receptors T-cell (CAR-T) therapy, and approaches for drug delivery across the BBB are emerging.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-26DOI: 10.1021/acs.molpharmaceut.4c00306
Abhijit Das, Barshana Bhattacharya, Sakuntala Gayen, Souvik Roy
Flavonoid-based organometallic complexes were revealed to be novel bioactive compounds. The taxifolin ruthenium-p-cymene nanoparticle (TaxRu-NPs) was produced in this study, and the toxicological assessment was done prior to in vivo chemotherapeutic research. Furthermore, the in vitro chemotherapeutic investigation used the A549 and NCI-H460 lung cancer cell lines. The in vitro study found that TaxRu-NPs induced apoptosis in lung cancer cells and hindered their ability to form colonies and migrate. The in vivo study showed that treatment with TaxRu-NPs restored the histological structure of a normal lung with less hyperplasia and lymphocytic infiltration. Furthermore, the treatment downregulated the angiogenic marker VEGF and the cell survival protein β-catenin and upregulated apoptotic markers like p53 and caspase-3. TaxRu-NPs treatment additionally raised the apoptotic index and decreased cancer cell growth. Finally, TaxRu-NPs effectively alleviate lung cancer by activating p53-mediated apoptosis and preventing angiogenesis and metastasis by decreasing the VEGF/β-catenin pathway.
{"title":"Suppression of Metastasis and Angiogenesis by Taxifolin Ruthenium-<i>p</i>-cymene Loaded PLGA Nanoparticles in Lung Carcinoma.","authors":"Abhijit Das, Barshana Bhattacharya, Sakuntala Gayen, Souvik Roy","doi":"10.1021/acs.molpharmaceut.4c00306","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00306","url":null,"abstract":"<p><p>Flavonoid-based organometallic complexes were revealed to be novel bioactive compounds. The taxifolin ruthenium-<i>p</i>-cymene nanoparticle (TaxRu-NPs) was produced in this study, and the toxicological assessment was done prior to in vivo chemotherapeutic research. Furthermore, the in vitro chemotherapeutic investigation used the A549 and NCI-H460 lung cancer cell lines. The in vitro study found that TaxRu-NPs induced apoptosis in lung cancer cells and hindered their ability to form colonies and migrate. The in vivo study showed that treatment with TaxRu-NPs restored the histological structure of a normal lung with less hyperplasia and lymphocytic infiltration. Furthermore, the treatment downregulated the angiogenic marker VEGF and the cell survival protein β-catenin and upregulated apoptotic markers like p53 and caspase-3. TaxRu-NPs treatment additionally raised the apoptotic index and decreased cancer cell growth. Finally, TaxRu-NPs effectively alleviate lung cancer by activating p53-mediated apoptosis and preventing angiogenesis and metastasis by decreasing the VEGF/β-catenin pathway.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142337297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p >B7-H3 has emerged as a promising target and potential biomarker for diagnosing tumors, evaluating treatment efficacy, and determining patient prognosis. Hu4G4 is a recombinant humanized antibody that selectively targets the extracellular domain of human B7-H3. In this study, we describe the radiolabeling of hu4G4 with the positron emission tomography (PET) emitter radionuclide zirconium 89 (<sup>89</sup>Zr) and evaluate its potency as an immuno-PET tracer for B7-H3-targeted imaging by comparing it <i>in vitro</i> and <i>in vivo</i> to [<sup>89</sup>Zr]Zr-DFO–DS-5573a using various models. The radiolabeled compound, [<sup>89</sup>Zr]Zr-desferrioxamine-hu4G4 ([<sup>89</sup>Zr]Zr-DFO-hu4G4), demonstrated a high radiochemical purity (RCP) of greater than 99% and a specific activity of 74 MBq/mg following purification. Additionally, it maintained stability in human serum albumin (HSA) and acetate buffer, preserving over 90% of its RCP after 7 days. Three cell lines targeting human B7-H3(U87/CT26<sub><i>-CD276</i></sub>/GL261<sub><i>-CD276</i></sub>) were used. Flow cytometry analysis indicated that the B7-H3-positive cells (U87/CT26<sub><i>-CD276</i></sub>/GL261<sub><i>-CD276</i></sub>) had a higher B7-H3 protein level with no expression in the B7-H3-negative cells (CT26<sub><i>-wt</i></sub>/GL261<sub><i>-wt</i></sub>) (<i>P</i> < 0.001). Moreover, the cellular uptake was 45.71 ± 3.78% for [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> cells versus only 0.93 ± 0.47% in CT26<sub><i>-wt</i></sub> cells and 30.26 ± 0.70% when [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> cells were blocked with 100× 8H9. The cellular uptake of [<sup>89</sup>Zr]Zr-DFO-hu4G4 was akin to that observed with [<sup>89</sup>Zr]Zr-DFO-DS-5573a with no significant differences (45.71 ± 3.78 % vs 47.07 ± 0.86 %) in CT26<sub><i>-CD276</i></sub> cells. Similarly, the CT26<sub><i>-CD276</i></sub> mouse model demonstrated markedly low organ uptake and elevated tumor uptake 48 h after [<sup>89</sup>Zr]Zr-DFO-hu4G4 injection. PET/CT analysis showed that the tumor-to-muscle (T/M) ratios were substantially higher compared to other imaging groups: 27.65 ± 3.17 in CT26<sub><i>-CD276</i></sub> mice versus 11.68 ± 4.19 in CT26<sub><i>-wt</i></sub> mice (<i>P</i> < 0.001) and 16.40 ± 0.78 when 100× 8H9 was used to block [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> mice (<i>P</i> < 0.01) at 48 h post-injection. Additionally, the tracer showed markedly high accumulation in the tumor region (22.57 ± 3.03% ID/g), comparable to the uptake of [<sup>89</sup>Zr]Zr-DFO–DS-5573a (24.76 ± 5.36% ID/g). A dosimetry estimation study revealed that the effective dose for [<sup>89</sup>Zr]Zr-DFO-hu4G4 was 2.96 × 10<sup>–01</sup> mSv/MBq, which falls within the acceptable range for further research in nuclear medicine. Collectively, these results indicated that [<sup>89</sup>Zr]Zr-DFO-hu4G4 was successfully fabricated and applied in B7-H3-targeted tumo
{"title":"Development of a Specifically Labeled 89Zr Antibody for the Noninvasive Imaging of Tumors Overexpressing B7-H3","authors":"Meng Zheng, Qingfeng Liu, Hua Zhang, Yanan Wang, Kaijie Zhang, Huiwen Mu, Fengqing Fu, Xueguang Zhang*, Yan Wang* and Liyan Miao*, ","doi":"10.1021/acs.molpharmaceut.4c0059710.1021/acs.molpharmaceut.4c00597","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00597https://doi.org/10.1021/acs.molpharmaceut.4c00597","url":null,"abstract":"<p >B7-H3 has emerged as a promising target and potential biomarker for diagnosing tumors, evaluating treatment efficacy, and determining patient prognosis. Hu4G4 is a recombinant humanized antibody that selectively targets the extracellular domain of human B7-H3. In this study, we describe the radiolabeling of hu4G4 with the positron emission tomography (PET) emitter radionuclide zirconium 89 (<sup>89</sup>Zr) and evaluate its potency as an immuno-PET tracer for B7-H3-targeted imaging by comparing it <i>in vitro</i> and <i>in vivo</i> to [<sup>89</sup>Zr]Zr-DFO–DS-5573a using various models. The radiolabeled compound, [<sup>89</sup>Zr]Zr-desferrioxamine-hu4G4 ([<sup>89</sup>Zr]Zr-DFO-hu4G4), demonstrated a high radiochemical purity (RCP) of greater than 99% and a specific activity of 74 MBq/mg following purification. Additionally, it maintained stability in human serum albumin (HSA) and acetate buffer, preserving over 90% of its RCP after 7 days. Three cell lines targeting human B7-H3(U87/CT26<sub><i>-CD276</i></sub>/GL261<sub><i>-CD276</i></sub>) were used. Flow cytometry analysis indicated that the B7-H3-positive cells (U87/CT26<sub><i>-CD276</i></sub>/GL261<sub><i>-CD276</i></sub>) had a higher B7-H3 protein level with no expression in the B7-H3-negative cells (CT26<sub><i>-wt</i></sub>/GL261<sub><i>-wt</i></sub>) (<i>P</i> < 0.001). Moreover, the cellular uptake was 45.71 ± 3.78% for [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> cells versus only 0.93 ± 0.47% in CT26<sub><i>-wt</i></sub> cells and 30.26 ± 0.70% when [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> cells were blocked with 100× 8H9. The cellular uptake of [<sup>89</sup>Zr]Zr-DFO-hu4G4 was akin to that observed with [<sup>89</sup>Zr]Zr-DFO-DS-5573a with no significant differences (45.71 ± 3.78 % vs 47.07 ± 0.86 %) in CT26<sub><i>-CD276</i></sub> cells. Similarly, the CT26<sub><i>-CD276</i></sub> mouse model demonstrated markedly low organ uptake and elevated tumor uptake 48 h after [<sup>89</sup>Zr]Zr-DFO-hu4G4 injection. PET/CT analysis showed that the tumor-to-muscle (T/M) ratios were substantially higher compared to other imaging groups: 27.65 ± 3.17 in CT26<sub><i>-CD276</i></sub> mice versus 11.68 ± 4.19 in CT26<sub><i>-wt</i></sub> mice (<i>P</i> < 0.001) and 16.40 ± 0.78 when 100× 8H9 was used to block [<sup>89</sup>Zr]Zr-DFO-hu4G4 in CT26<sub><i>-CD276</i></sub> mice (<i>P</i> < 0.01) at 48 h post-injection. Additionally, the tracer showed markedly high accumulation in the tumor region (22.57 ± 3.03% ID/g), comparable to the uptake of [<sup>89</sup>Zr]Zr-DFO–DS-5573a (24.76 ± 5.36% ID/g). A dosimetry estimation study revealed that the effective dose for [<sup>89</sup>Zr]Zr-DFO-hu4G4 was 2.96 × 10<sup>–01</sup> mSv/MBq, which falls within the acceptable range for further research in nuclear medicine. Collectively, these results indicated that [<sup>89</sup>Zr]Zr-DFO-hu4G4 was successfully fabricated and applied in B7-H3-targeted tumo","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142403697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-25DOI: 10.1021/acs.molpharmaceut.4c0102610.1021/acs.molpharmaceut.4c01026
Malcolm Lim*,
{"title":"Voices in Molecular Pharmaceutics: Meet Dr. Malcolm Lim, Who Advances Treatment for Brain Metastases with Targeted Radiopharmaceuticals","authors":"Malcolm Lim*, ","doi":"10.1021/acs.molpharmaceut.4c0102610.1021/acs.molpharmaceut.4c01026","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c01026https://doi.org/10.1021/acs.molpharmaceut.4c01026","url":null,"abstract":"","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142403609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-24DOI: 10.1021/acs.molpharmaceut.4c0064310.1021/acs.molpharmaceut.4c00643
Xiaohui Wang*, Zhijian Han, Jun Zhang, Ming Chen and Wenbo Meng*,
Heat shock protein 90 (Hsp90) is a promising target for cancer therapy and imaging. Accurate detection of Hsp90 levels in tumors via noninvasive PET imaging might be beneficial for management. To achieve this, the precursor compound Dimer-Sansalvamide A (Dimer-San A) was PEGylated and modified by conjugating it with the bifunctional chelator 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). The 18F-labeled PEGylated Dimer-SanA decapeptide (18F-PEGylated San A) was completed within 30 min using a two-step process. In vitro stability and specificity were assessed, including competition studies with the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG). MicroPET imaging was performed on PL45 tumor-bearing mice to evaluate probe accumulation and tumor-to-muscle ratios. Biodistribution studies determined the route of excretion. The probe resulted in a radiochemical yield of 23.11% with a purity exceeding 95%. In vitro, 18F-PEGylated San A exhibited high stability and selectively accumulated in Hsp90-positive PL45 cells, with binding effectively blocked by the Hsp90 inhibitor 17AAG, confirming its specificity. MicroPET imaging of PL45 tumor-bearing mice showed significant probe accumulation in tumor tissues at 1 and 2 h postinjection (4.06 ± 0.30 and 3.72 ± 0.61%ID/g, respectively), with optimal tumor-to-muscle ratios observed at 2 h postinjection (6.09 ± 1.92). While 18F-PEGylated San A demonstrates enhanced water solubility, as indicated by increased kidney uptake relative to liver accumulation. The study successfully incorporated PEG units to create the novel probe 18F-PEGylated San A targeting to Hsp90 without affecting its targeting capability, aimed at improving the pharmacokinetics and PET imaging of Hsp90 expression noninvasively.
{"title":"Development and Preclinical Evaluation of 18F-Labeled PEGylated Sansalvamide A Decapeptide for Noninvasive Evaluation of Hsp90 Status in Pancreas Cancer","authors":"Xiaohui Wang*, Zhijian Han, Jun Zhang, Ming Chen and Wenbo Meng*, ","doi":"10.1021/acs.molpharmaceut.4c0064310.1021/acs.molpharmaceut.4c00643","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.4c00643https://doi.org/10.1021/acs.molpharmaceut.4c00643","url":null,"abstract":"<p >Heat shock protein 90 (Hsp90) is a promising target for cancer therapy and imaging. Accurate detection of Hsp90 levels in tumors via noninvasive PET imaging might be beneficial for management. To achieve this, the precursor compound Dimer-Sansalvamide A (Dimer-San A) was PEGylated and modified by conjugating it with the bifunctional chelator 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). The <sup>18</sup>F-labeled PEGylated Dimer-SanA decapeptide (<sup>18</sup>F-PEGylated San A) was completed within 30 min using a two-step process. <i>In vitro</i> stability and specificity were assessed, including competition studies with the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG). MicroPET imaging was performed on PL45 tumor-bearing mice to evaluate probe accumulation and tumor-to-muscle ratios. Biodistribution studies determined the route of excretion. The probe resulted in a radiochemical yield of 23.11% with a purity exceeding 95%. <i>In vitro</i>, <sup>18</sup>F-PEGylated San A exhibited high stability and selectively accumulated in Hsp90-positive PL45 cells, with binding effectively blocked by the Hsp90 inhibitor 17AAG, confirming its specificity. MicroPET imaging of PL45 tumor-bearing mice showed significant probe accumulation in tumor tissues at 1 and 2 h postinjection (4.06 ± 0.30 and 3.72 ± 0.61%ID/g, respectively), with optimal tumor-to-muscle ratios observed at 2 h postinjection (6.09 ± 1.92). While <sup>18</sup>F-PEGylated San A demonstrates enhanced water solubility, as indicated by increased kidney uptake relative to liver accumulation. The study successfully incorporated PEG units to create the novel probe <sup>18</sup>F-PEGylated San A targeting to Hsp90 without affecting its targeting capability, aimed at improving the pharmacokinetics and PET imaging of Hsp90 expression noninvasively.</p>","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142403960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}