Pharmaceutical Research

Mehrotra, Nidhi; Manchikanti, Padmavati

doi: 10.1007/s11095-025-03988-zpmid: 41372687

Although India ranks among the largest producers and exporters of human vaccines, the delay in launch of newer therapeutics is larger than the average in G20 nations. This study aims to assess the stakeholders’ perspective on the coherence, clarity, and operational impact of the New Drugs and Clinical Trial (NDCT) Rules 2019. The analysis revealed a nuanced appraisal of the NDCT Rules wherein three major clusters of challenges faced by pharmaceutical companies are regulatory, operational, and systemic. The regulatory barriers encompass prolonged approval timelines, ambiguous legal mechanisms, fragmented oversight across agencies. Operational factors include uneven distribution of trial sites, inadequate site capacity, ethics committee delays. Systemic challenges centred on non-uniform trial protocols, inconsistent documentation practices and absence of harmonized operational guidelines.

Martins, Frederico Severino; Bottos, Gabriel; Silva Hecktheuer, Amanda; Luiz Borges, Leonardo

doi: 10.1007/s11095-025-03983-4pmid: 41350967

Biotechnology is becoming a key driver of innovation in health, agriculture, industry, and sustainability worldwide. Brazil has made significant progress in this field, thanks to its strong research institutions, diverse ecosystems, and active pharmaceutical and agribusiness sectors. However, despite these strengths, the country still faces critical barriers that limit its global competitiveness in biotechnology. These include a lack of coordination between academia and industry, limited early-stage investment, underdeveloped infrastructure, regulatory delays, and low levels of internationalization. This article provides a comprehensive overview of the biotechnology landscape in Brazil. It analyzes the global context, highlights the country’s current position, and identifies gaps and opportunities across the biopharmaceutical value chain. Drawing from international benchmarks and national data, the paper proposes five strategic directions to strengthen Brazil’s biotechnology ecosystem: (1) fostering translational science and innovation; (2) expanding access to capital and reducing investment risk; (3) improving infrastructure through shared platforms; (4) modernizing regulatory frameworks; and (5) positioning Brazil in global markets through its unique strengths, such as biodiversity, public health expertise, and agricultural leadership. The article concludes that Brazil has the scientific foundation and market potential to become a global player in biotechnology. However, realizing this potential will require coordinated efforts from government, industry, and academia. With the right policies and investments, Brazil can turn scientific advances into innovative solutions that benefit both its population and the global bioeconomy.

Sood, Abhilasha; Mehrotra, Arpit

doi: 10.1007/s11095-025-04004-0pmid: 41507551

BackgroundMitochondria besides being the powerhouse of the cell are also involved in performing a multitude of critical cellular functions. Any failure in maintenance of these organelles is implicated in multiple human pathologies, including neurodegenerative disorders. Over the past two decades, significant efforts have been made to investigate the pharmacodynamic propensity of various potential compounds, which could be engaged as efficient therapeutic approach in modulating mitochondrial dynamics during neuronal dysfunctions.MethodThis review comprehensively overviews the contribution of potential compounds that could be employed as mitochondrial medicine in reversing neurological pathologies, with special focus on their significant roles as: metabolic antioxidants, conjugated molecules for mitochondrial function modulation, mitochondrial targeted peptides, optogenetic based induction of the mitochondria, potential mitochondrial biomarkers and other advanced transportation systems for mitochondrial delivery to brain.Results and DiscussionThe manuscript discusses the mechanism of action of potential compounds (natural and pharmacologically synthesized), and other advance approaches that could efficiently modulate mitochondrial machinery in terms of regulating mitochondrial biogenesis, mitophagy, bioenergetics pathways, oxidative stress, calcium homeostasis and mitochondrial DNA stability.ConclusionThe optimal maintenance of mitochondrial dynamics offered by variety of mitochondria targeting compounds highlights their prospective value for considering them as futuristic neurotherapeutic agents, which could be considered in managing neurodegenerative conditions.

Wu, Haomin; Fan, QinXi; Zhang, Zheng; Ji, Yuanhui

doi: 10.1007/s11095-025-03980-7pmid: 41266728

BackgroundTherapeutic proteins are playing an increasingly important role in marketed drugs and clinical candidates. However, their development still faces major challenges, particularly aggregation.ObjectivesThis review explores the recent advancements, current limitations, and future directions of new research methods for therapeutic proteins.ResultsCharacterization techniques identify aggregation tendencies and elucidate underlying mechanisms, while computational chemistry provides microscopic insights into the aggregation process. Theoretical modeling and machine learning offer tools for predicting protein stability, enabling high-throughput screening in early formulation development.ConclusionFostering interdisciplinary collaboration will be essential. The integration of diverse approaches offers a more comprehensive understanding of protein aggregation and unlocks new opportunities for innovation in protein formulation development.

Basu, Koushiki; Chelagamsetty, Venkata S.; Ruiz-Avila, Veronica A.; Li, Tonglei

doi: 10.1007/s11095-025-03981-6pmid: 41298984

Nanoparticles (NPs), due to their small size and large surface area, have advanced their use as drug carriers for delivering various therapeutic molecules. When entering biological environments, nanoparticles typically adsorb proteins, forming a surface layer known as a protein corona that significantly affects the biological and therapeutic functions of a delivery system. Understanding and predicting protein adsorption is essential for optimizing nanoparticle design in drug delivery, diagnostics, and therapy. Machine learning and deep learning (ML/DL) offer promising methods for designing nanoparticles with specific properties, particularly given recent advancements in computation and nanoparticle analysis. This review explores ML/DL studies of nanoparticle-protein interactions and emphasizes the popularity of Random Forest (RF) and Deep Learning (DL) models in predicting protein corona compositions. RF models are highly valued for managing high-dimensional data and offering interpretability, which helps identify key NP features influencing protein adsorption. Conversely, DL excels at modeling non-linear relationships and detecting subtle interaction patterns. While most current research focuses on protein coronas, future models may also include other biocorona components. This is particularly relevant for soft materials, such as lipid nanoparticles (LNPs), which are now approved for delivering mRNA and peptide-based vaccines. Our findings underscore the need for advanced modeling techniques and high-quality, diverse experimental data to drive innovations in nanomedicine. Combining RF and DL approaches leverages their complementary strengths to overcome the challenge of limited experimental data and further improve NP designs for biomedical use.Graphical Abstract[graphic not available: see fulltext]

Nguyen-Thi, Phuong-Trang; Nguyen, Nhat Thang Thi; Le, Huong Thuy; Nguyen, Thuy Trang; Van Vo, Giau

doi: 10.1007/s11095-025-03995-0pmid: 41429993

Microneedles (MNs) provide a minimally invasive and efficient platform for transdermal drug delivery, offering precise control over dosage and release kinetics. Recent advances in dual-drug delivery using dissolvable MNs have focused on optimizing structural design, material composition, and programmable release mechanisms. Dual-layer or core–shell MN configurations allow spatial and temporal separation of drugs, while stimuli-responsive polymers enable release in response to physiological cues such as glucose concentration, pH, or reactive oxygen species (ROS). This review summarizes emerging strategies for co-delivery through dissolving MNs, emphasizing how design parameters including morphology, materials, and nanoformulations influence mechanical performance and drug-release profiles. Applications in cancer, diabetes, wound healing, and inflammatory diseases are highlighted. For example, a dual-drug MN co-loaded with an anti-PD-L1 antibody and 1-methyl-D,L-tryptophan (1-MT) achieved prolonged intratumoral retention and enhanced antitumor efficacy. Similarly, MNs incorporating MnSH nanozymes and polymyxin B demonstrated synergistic antibacterial and pro-angiogenic effects in wound models. The integration of nanocarriers and responsive polymers has expanded the therapeutic potential of MN-based systems, enabling precise, localized, and sustained co-delivery of active agents. Finally, current challenges including large-scale manufacturing, reproducibility, clinical validation, and regulatory approval are discussed to outline future directions for translating MN-based dual-drug delivery into clinical practice.

Feng, Chunlai; Deng, Wen; Cai, Min; Hu, Yujiao; Liang, Wenyan; Dong, Hangyu; Rui, Mengjie

doi: 10.1007/s11095-025-03990-5pmid: 41372692

ObjectiveBioactive peptides derived from natural or synthetic sources have shown significant potential for cancer therapy; however, their clinical application is often limited by poor tumor selectivity and systemic toxicity. In this work, we aim to develop a multifunctional antitumor peptide with enhanced tumor- targeting capability and therapeutic efficacy.MethodsA cationic antimicrobial peptide (AMP, (KKWW)2 K-NH2) was chemically modified via cysteine-mediated conjugation with 4-vinylphenylboronic acid, yielding the phenylboronic acid-conjugated AMP (PBA-AMP). Molecular dynamics (MD) simulations were performed to evaluate peptide-membrane interactions. Cellular uptake, cytotoxicity, and in vivo tumor-targeting and antitumor efficacy were assessed using MCF-7 cells and 4T1 tumor-bearing mice.ResultsMD simulations demonstrated that PBA-AMP exhibited rapid and stable binding to tumor cell membranes, maintaining consistent membrane interactions over a 50 ns simulation. Cellular studies revealed enhanced cellular uptake and increased cytotoxicity of PBA-AMP against MCF-7 breast cancer cell line (IC50 = 38.46 μM) compared to naive AMP (IC50 = 110 μM). In vivo imaging confirmed selective and prolonged tumor accumulation of PBA-AMP. Treatment with PBA-AMP significantly suppressed tumor growth in 4T1 tumor-bearing mice without observable systemic toxicity.ConclusionsThis study presents a rational design strategy for engineering tumor-selective, microenvironment-responsive therapeutic peptides. PBA-AMP represents a promising candidate for targeted cancer therapy, offering improved efficacy and reduced off-target effects.

Kurosawa, Toshiki; Ohtsuki, Sumio; Ishikawa, Hiroshi; Schroten, Horst; Schwerk, Christian; Kubo, Yoshiyuki; Deguchi, Yoshiharu

doi: 10.1007/s11095-025-03987-0pmid: 41372689

PurposeHuman choroid plexus papilloma (HIBCPP) cells derived from choroid plexus papilloma in the lateral ventricle are considered suitable model cells for the blood-cerebrospinal fluid barrier (BCSFB). Therefore, in this study, the transporters expressed in HIBCPP cells were identified, and the functions of representative transporters were evaluated.MethodsTransporter protein and mRNA expression were analyzed in HIBCPP cells using quantitative proteomics and quantitative reverse transcription-PCR. Transporter functions were evaluated by cellular uptake and transcellular transport studies using typical substrates.ResultsThe following solute carrier (SLC) and ATP-binding cassette (ABC) transporters involved in drug and nutrient transport were expressed in HIBCPP cells: glucose transporter 1 (GLUT1), monocarboxylate transporter 1 (MCT1), L-type amino acid transporter 1 (LAT1), cationic amino acid transporter 1 (CAT1), glutamate transporter (GLAST), SLC35F2, multidrug resistance-associated protein 1 (MRP1), and breast cancer resistance protein (BCRP). Furthermore, the mRNA of organic anion/cation transporters, such as organic anion transporting polypeptide 1B3 (OATP1B3) and plasma membrane monoamine transporter (PMAT), were detected. Additionally, uptake of representative substrates of the SLC transporters LAT1, CAT1, GLAST, GLUT1, MCT1, and SLC35F2 in HIBCPP cells occurred in a time- and temperature-dependent manner and decreased in the presence of specific inhibitors. Furthermore, the representative substrates of these transporters were transported in a symmetric or asymmetric manner through HIBCPP cells between the apical (cerebrospinal fluid [CSF]) and basolateral (blood) sides.ConclusionVarious nutrient and drug transporters are functionally expressed in HIBCPP cells. Therefore, HIBCPP cells could serve as a useful human BCSFB model to evaluate nutrient and drug transport between the CSF and blood.

Zhou, Simon; Rajanayake, Krishani Kumari; He, Miao; Wen, Bo; Lkhagva, Ankhbayar; Yap, Ernie; Sun, Duxin; Cross, Jennifer; Engelke, Kory; Huizinga, Robert B.

doi: 10.1007/s11095-025-03943-ypmid: 41507552

BackgroundCalcineurin inhibitors (CNIs) are immunosuppressive agents that inhibit calcineurin (CN) and are recommended for the treatment of lupus nephritis (LN). In clinical trials, differences in the safety profiles of CNIs have been observed. Emerging data suggests that small molecule therapeutics may be differentially distributed and retained within organ tissues, potentially explaining these safety profile disparities.MethodsThis study investigated the renal distribution and retention of the CNIs voclosporin (VCS), tacrolimus (TAC), and cyclosporine A (CSA) in CD-1 mice using matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-MSI).ResultsDistinct patterns of the distribution and retention of these compounds were observed. VCS showed a moderate cortical distribution, peaking between 15- and 30 min post administration, and was then rapidly excreted with minimal renal retention observed by 2 h post-dosing. In contrast, CSA exhibited a diffuse, persistent distribution in renal structures for up to 4 h post-dosing. TAC showed a diffuse distribution pattern, with retention observed in the renal medulla for up to 7 h post-dosing.ConclusionsThese data indicate that CNIs display different renal handling profiles. The shorter duration of renal retention of VCS demonstrated in the healthy mice indicates a differentiated profile compared to the other CNIs. Further research on the body-wide tissue distribution and renal handling of TAC, VCS and CSA in humans will aid in delineating the distinct clinical profiles of CNIs and optimize their use in treating immune disorders.

Wang, Yuncheng; Stevens, Maxwell T.; Wang, Trixie; Alreja, Adit B.; Nelson, Daniel C.; Britton, Warwick J.; Chan, Hak-Kim

doi: 10.1007/s11095-025-03942-zpmid: 41526528

PurposeInhalable liquid formulation of endolysins represents a promising alternative to conventional antibiotics. Dry powder formulations offer improved stability for endolysin pulmonary delivery. This study aimed to evaluate the efficacy of an inhalable dry powder or liquid formulation of endolysin Cpl-1 alone and to compare it with liquid combinations of Cpl-1 with either gentamicin or endolysin Pal in a murine model of S. pneumoniae lung infection.MethodsA dry powder formulation of Cpl-1 was produced via spray drying, while liquid formulations were prepared by dissolving Cpl-1, or in combination with gentamicin or endolysin Pal in liquid. The droplet size distribution of aerosolized formulations was also characterized. Mice were intratracheally infected with S. pneumoniae and treated with either powder or liquid formulations. The bacterial load in respiratory system was assessed 26 h post-infection. The stability and activity of Cpl-1 in BALF were also evaluated ex vivo.ResultsA single dose of Cpl-1 powder formulation or Cpl-1 liquid formulation (40 µg/mouse) reduced pulmonary bacterial load by approximately 1 log10. Importantly, the combination of Cpl-1 and Pal in liquid form resulted in a synergistic 2.0 log10 reduction, significantly greater than either endolysin alone, while combining Cpl-1 with gentamicin did not enhance antibacterial activity. Ex vivo assays confirmed that Cpl-1 retained full enzymatic activity after incubation in BALF.ConclusionThis proof-of-principle study demonstrated that inhalable endolysin liquid and powder formulations could potentially be used to treat bacterial lung infections. Moreover, the combination of multiple endolysins could increase antimicrobial activity over endolysin monotherapy.