Pharmaceutics doi: 10.3390/pharmaceutics18091112

Authors: Yohan Seo Chul Soon Park

Major depressive disorder remains a leading cause of disability worldwide, and the limited efficacy and delayed onset of conventional antidepressants have intensified interest in the microbiota–gut–brain axis as a source of therapeutic targets. Microbiota-associated candidates—short-chain fatty acids, bile acid and tryptophan metabolites, and neuroactive amines—show mood-relevant activity, yet almost none has reached the clinic. This review reframes that gap as a delivery problem. Rather than treating entry into the central nervous system as a universal requirement, we distinguish strategies intended for local intestinal, peripheral systemic, and direct central action, and we argue that delivery is a major but not exclusive translational bottleneck. We outline the barriers these agents face—upper gastrointestinal loss, poor colonic targeting, rapid metabolite turnover, first-pass exposure, and the blood–brain barrier—and synthesize delivery strategies across two fronts. Colon-targeted systems are technically established but have been validated for non-depression indications, whereas brain-directed approaches—bacterial extracellular vesicles, detoxified membrane-coated carriers, receptor-mediated transcytosis, and intranasal routes—reach the brain mainly in selected preclinical models. We foreground a paradox: microbial extracellular vesicles are at once one of the better-documented bio-derived routes for brain exposure in preclinical studies and prominent drivers of neuroinflammation, which defines a risk–opportunity continuum. We close with a route-specific validation roadmap encompassing quantitative exposure, target engagement, chronic efficacy, and safety.

Pharmaceutics doi: 10.3390/pharmaceutics18091111

Authors: Diana Constanța Pelea Laura Maria Endres Laura Maghiar Teodor-Andrei Maghiar Mădălin Florin Ganea Csaba Nagy Corina Moisa Gabriela Ciavoi Georgiana Ioana Potra Cicalau Olimpia-Daniela Frenț Mariana Ganea

Background/Objectives: Antimicrobial resistance has renewed interest in plant extracts, essential oils and honey as topical adjuvants, yet antimicrobial potency, host-tissue safety and formulation performance are usually reported in separate bodies of literature, and the material tested is often characterized only superficially. This structured narrative review aimed to integrate these dimensions into a single decision-oriented pathway. Methods: The Web of Science Core Collection was searched using seventeen predefined strings across fourteen thematic domains, restricted to English-language articles and reviews from 2021 to 2026, with supplementary PubMed/MEDLINE searches, hand-searching, and normative documents from the issuing organizations. Results: The framework comprises seven decision domains: pre-analytical standardization of each matrix class; antibacterial and antifungal testing with matrix-specific controls; biofilm endpoints named according to what they measure; strain selection tiered from reference to clinical and resistant isolates; host safety from monolayer cytotoxicity to reconstructed human epidermis; selectivity, expressed per microorganism and extended to the resident cutaneous microbiota; and formulation performance, with permeation interpreted against a retention rather than a maximization target. Resistance under sub-inhibitory exposure forms an advanced stage. Conclusions: Progression is governed by proposed Go, Conditional Go and No-Go criteria, supported by a matrix-specific control table and a minimum reporting checklist. The framework has not been prospectively validated, and no universal selectivity threshold is proposed.

Pharmaceutics doi: 10.3390/pharmaceutics18091110

Authors: Jinhwan Shin Eu-min Kim Jun-hong Jang Seok-won Jee Sang-hoon Kim Seonggwan Yu Minguen Yoon Daniel Craig Ryan Swoyer Sandip Patel Praveen Alamuri Albert Price Rashmi Ravichandran Lauren Carter Sammaiah Pallerla

Background/Objectives: The rapid emergence of SARS-CoV-2 variants that evade neutralizing antibodies underscores the need for new antiviral biologics that integrate precise molecular design with scalable, cost-effective manufacturing. Computationally designed miniproteins targeting the spike protein’s receptor-binding domain (RBD) offer a promising alternative to monoclonal antibodies because of their small size, high thermal stability, and compatibility with microbial expression systems. Here, we detail the full development and cGMP manufacturing of IPD-52520, a novel antiviral miniprotein produced using an optimized E. coli platform. Methods: Two candidate miniproteins, a homotrimeric construct (Trimer, IPD-52520, 17 kDa) and a tandem fusion construct (Daisy, IPD-52521, 25 kDa), were evaluated in parallel through systematic optimization of strain selection, media formulation, fed-batch fermentation, inclusion-body solubilization, refolding, and chromatographic purification. The leading candidate was scaled from 5 L to 50 L under cGMP conditions, and biophysical and stability analyses were conducted to support nonclinical and Phase 1 clinical development. Results: The Trimer was selected as the lead molecule based on superior preclinical efficacy, favorable pharmacokinetics, and higher volumetric yields. The optimized process yields about 2 g/L of purified protein at over 90% purity. Scale-up from 5 L to 50 L demonstrated excellent batch consistency across six independent runs. Biophysical studies confirmed a well-folded, predominantly alpha-helical trimer (Tm = 73.4 °C; polydispersity = 1.005) with an intact primary structure and strong target binding (KD < 1 pM). Stability studies show the drug remains stable at 2–8 °C for at least 12 months, with ongoing long-term studies. Conclusions: These findings demonstrate that computationally designed antiviral miniproteins can be translated into scalable biologics and establish a platform for rapid therapeutic development against current and future pandemics.

Pharmaceutics doi: 10.3390/pharmaceutics18091109

Authors: Aurane Lecouffe Thomas Bouillet Bryan Thiroux Amélie Degraeve Anaïs-Camille Vreulx Romain Magnez Angelo Scuotto Christophe Barnier-Quer Didier Betbeder

Background/Objectives: Toxoplasma gondii is a globally distributed parasite responsible for significant morbidity in both humans and animals. VXN-Toxo, is an intranasal vaccine based on maltodextrin nanoparticles formulated with inactivated T. gondii parasites. A vaccination campaign conducted worldwide in zoological parks demonstrated high efficacy across multiple species and geographical regions. These findings suggest that VXN-Toxo may induce broad cross-reactive immunity against T. gondii strains circulating in various regions. Methods: To further characterize the immune mechanisms, we evaluated the cellular immune response induced by VXN-Toxo in C57BL/6 mice. Following vaccination, splenocytes were stimulated with antigens derived from multiple T. gondii strains, representing the major haplogroups (types I, II, III, and atypical strains). Results: ELISPOT analysis demonstrated that VXN-Toxo induced strong antigen-specific T cell responses, characterized by robust IFN-γ and IL-17 production upon stimulation with both homologous and heterologous antigens. Flow cytometry analysis further revealed the activation of both CD4+ and CD8+ T cells, with the notable presence of IFN-γ–producing CD8+ central and effector memory cells and CD4+ effector memory T cells. Conclusions: Altogether, these results indicate that VXN-Toxo induces a broad, T cell–mediated immune response with cross-reactive properties and highlight its potential as a promising vaccine candidate for both human and veterinary applications.

Pharmaceutics doi: 10.3390/pharmaceutics18091108

Authors: Lu Lu Yige Qiu Jiayu Wu Wei Dou Jing Yang Bo Zhang

RNA therapeutics offer considerable potential for cancer treatment. Their therapeutic application, however, remains limited by rapid degradation, inefficient cellular uptake, and restricted intracellular release. Extracellular vesicles (EVs) are cell-derived membrane vesicles that have been exploited as promising vehicles for drug delivery due to their high biocompatibility and low immunogenicity, whereas liposomes and lipid nanoparticles provide tunable lipid composition and efficient loading of exogenous nucleic acids. Combining these carriers has led to EV–lipid hybrid systems designed to integrate their complementary properties. This review summarizes recent advances in EV–lipid hybrids for cancer therapy and organizes the reported systems according to their structural architecture and preparation. EV–liposome fusion hybrids, EV–lipid nanoparticle hybrids, and EV membrane-integrated lipid nanocarriers are discussed in relation to their RNA-loading strategies and representative therapeutic designs. The review also examines the key processes involved in functional RNA delivery and summarizes representative applications across different cancer types. Challenges associated with safety evaluation are also discussed, together with future directions for clinical translation. Overall, EV–lipid hybrids represent a promising strategy for RNA-based cancer therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091107

Authors: Jun Liu Shuang Liu Jianxiong Wei Zifang Ding Xiaodan Yan Jin Sun Shujun Wang Shanhu Li Yuanqing Li

Background: The oral application of adenovirus is hindered by its poor in vitro storage stability and rapid degradation by gastric acid. To address this, a biomimetic oral adenovirus delivery system (CSP-AdV@LYO) was constructed based on three key properties of natural chrysanthemum sporopollenin (CSP): chemical inertness, intelligent “acid-shrinking/alkali-swelling” responsiveness, and mucosal adhesion via its spike structures, aiming to enhance oral stability and delivery efficiency. Methods: First, low-allergenic chrysanthemum pollen was screened using proteomics and a zebrafish allergy model. High-purity sporopollenin (SPO) was then extracted via an acidolysis method, followed by systematic characterization of its morphology, particle size, zeta potential, contact angle, and reversible acid-shrinking/alkali-swelling behavior. Subsequently, a CSP-AdV@LYO formulation was prepared by optimizing a cryoprotectant formulation (sucrose:gelatin = 1:1) and a vacuum loading process. Its protective and release properties were evaluated in vitro using simulated gastric and intestinal fluids, and its long-term stability was assessed. Further in vivo studies in mice assessed its intestinal colonization efficiency. The adhesion mechanism of the sporopollenin spike structures was investigated through mucosal retention experiments. Results: Mucosal retention experiments confirmed that the spike structures on the sporopollenin surface enhanced retention by approximately 3-fold through mechanical interlocking compared to smooth particles. In long-term stability tests, the viral genome copy number retention rate was improved more than 10-fold compared to the virus stock solution. The system enabled a steady and controlled release of the virus in simulated intestinal fluid, with the released virus maintaining its infectivity. In vivo studies demonstrated that CSP-AdV@LYO promoted efficient intestinal colonization and reduced acute mortality from 75% (AdV@LYO group) to 25%. Conclusion: By leveraging the unique physicochemical properties of chrysanthemum sporopollenin, this study successfully developed a biomimetic oral delivery system for live adenovirus that provides gastric acid protection, intelligent pH-responsive release, and mucosal adhesion. This system significantly enhances the oral stability and intestinal delivery efficiency of adenovirus while reducing systemic exposure risks. It offers a novel biomimetic strategy for the oral delivery of adenovirus and other biological macromolecules.

Pharmaceutics doi: 10.3390/pharmaceutics18091105

Authors: Qingqing Xu Lisong Sheng Hui Zhao Lingyun Du Jingjing Wei Huijie Zhang Tianyu Zhang Huanhuan Zhang Chunhong Zhang Rong Sun

Background/Objectives: Psoriasis is a chronic inflammatory skin disease driven by Th17/Treg imbalance. Colquhounia Root Tablet (CRT), derived from Tripterygium hypoglaucum, has shown clinical potential for psoriasis, but its mechanisms remain unclear. This study aimed to evaluate the anti-psoriatic effects of CRT and elucidate its underlying mechanisms. Methods: Anti-psoriatic activity was evaluated in an IMQ-induced psoriasis-like mouse model. Mice received oral CRT at 0.085, 0.17, or 0.35 g/kg daily from days 2 to 8. Immune-cell populations were analyzed by flow cytometry. Bone marrow-derived dendritic cells (BMDCs) were used for in vitro studies. Network pharmacology, transcriptomics, molecular docking, and experimental validation were integrated to explore the mechanisms. Results: CRT dose-dependently ameliorated psoriasiform dermatitis and reduced Th17/Treg ratio while inhibiting CD11c+MHC II+ DC activation in vivo. In vitro, CRT suppressed R848-induced BMDC maturation and inhibited p65/IκBα phosphorylation. Transcriptomic analysis revealed modulation of TNF, NF-κB, IL-17, and JAK-STAT pathways. Molecular docking predicted the strong binding of multiple CRT compounds to RELA. Conclusions: CRT exerts anti-psoriatic effects in a murine model with concurrent modulation of NF-κB-related DC maturation and Th17/Treg correction, suggesting a potential immunomodulatory mechanism requiring further causal validation.

Pharmaceutics doi: 10.3390/pharmaceutics18091106

Authors: Grzegorz Szynkaruk Julia Kerner Ahata Nelipovich Kacper Osuch Maria Miotk Agata Bartkowiak Joanna Sobiak

Background/Objectives: Saliva may provide a non-invasive alternative to blood sampling for therapeutic drug monitoring (TDM) of mycophenolic acid (MPA) in children. This study assessed MPA and, secondarily, mycophenolic acid glucuronide (MPAG) in non-centrifuged Salivette® devices to determine an acceptable interval between saliva collection and laboratory processing. Methods: Phosphate-buffered saline (PBS), artificial saliva, and saliva from five healthy adult volunteers were externally spiked with MPA and MPAG at 5 and 500 ng/mL and applied to Salivette® cotton swabs. Samples were stored for 12, 24, and 48 h at 22 °C and 6 °C and analyzed by liquid chromatography-tandem mass spectrometry. Percentage deviations from nominal concentration within ±15% were considered acceptable. Results: In PBS, both analytes met the acceptance criterion for up to 24 h under both conditions. In artificial saliva, both met the criterion for at least 24 h and, under some conditions, for 48 h. In human saliva, all volunteer-level MPA results met the criterion after 12 h at both temperatures, whereas MPAG did not consistently meet it. Mean MPA deviations across volunteers ranged from −10.8 to 1.9% at 22 °C and from −12.3 to 2.4% at 6 °C. Conclusions: MPA met the acceptance criterion after 12-h in non-centrifuged Salivette® devices, supporting the feasibility of a 12-h pre-centrifugation interval under comparable conditions. Prompt centrifugation and processing are advisable when MPAG determination is required. Confirmation using incurred post-dose saliva from pediatric patients receiving mycophenolate mofetil is required.

Pharmaceutics doi: 10.3390/pharmaceutics18091104

Authors: Tünay Karan Çağrı Çağlar Sinmez Sevgi Durna Daştan Murat Çakir Mücahit Seçme René van den Hoven

A wound is defined as disruption or destruction of tissue integrity. In order to support healing in wound management, a good wound bed free of necrotic tissue and infection is desired, but intensive chemical antiseptics will cause cell destruction and delay healing. In order for wound healing to be rapid, the nature and contamination of the wound should be taken into consideration and appropriate methods should be utilized. Today, many types of algae are frequently preferred as an alternative to medicine and are the subject of research. Since the metabolites contained in algae display several notable biological activities such as antimicrobial, anti-inflammatory, and antioxidant, they are a good option in wound treatment. Algae contain pigments, peptides, fatty acids, and polysaccharides that are crucial for wound healing. These compounds play vital roles at all stages of the healing process by accelerating cell proliferation, promoting collagen deposition, scavenging reactive oxygen species (ROS), and regulating key inflammatory cytokines. Furthermore, their unique physical and functional properties enable the development of novel bio-inspired wound dressings, hydrogels, and drug-delivery scaffolds. This review discusses the bioactive metabolites found in algae that are effective in wound healing.

Pharmaceutics doi: 10.3390/pharmaceutics18091103

Authors: Lara Marques Nuno Vale

Background/Objectives: Interindividual pharmacokinetic (PK) variability remains a daunting challenge for effective and safe drug therapy. Despite the widespread use of direct oral anticoagulants (DOACs) and calcium channel blockers (CCBs), a substantial number of adverse drug reactions have been reported for both classes. Herein, this study aimed to assess and analyze the PK variability of DOACs and CCBs across diverse clinical and demographic profiles under both single- and multiple-dose conditions. Methods: A PubMed search identified clinical PK studies reporting maximum plasma concentration (Cmax) and/or area under the concentration-time curve (AUC). The coefficient of variation (CV%) was calculated and used as a measure of PK variability. A CV% < 40% indicated low-to-moderate variability, and a CV% > 40% was defined as high variability. Results: A total of 264 studies were included following systematic screening, and the dataset was further characterized according to population features and clinical context. Among DOACs, edoxaban exhibited the lowest PK variability, whereas dabigatran showed the highest. CCBs demonstrated a broad variability spectrum, ranging from predictable agents (amlodipine and felodipine) to highly variable compounds (nisoldipine, isradipine, nimodipine, diltiazem, and verapamil). Studies evaluating drug–drug interactions, ethnicity, and specific drug-related factors were associated with increased PK variability. Conclusions: These findings suggest that fixed-dose strategies may not be universally appropriate for DOACs and CCBs, particularly in high-risk subgroups where altered exposure may lead to sub- or supratherapeutic concentrations and compromise clinical outcomes. Therefore, clinicians should avoid evaluating individual risk factors in isolation and instead consider the patient’s complete profile when selecting and adjusting pharmacotherapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091102

Authors: Hiroki Kato Xuhao Huang Erina Hilmayanti Yuichiro Kadonaga Kazuhiro Ooe Masashi Murakami Kazuya Kabayama Kazuko Kaneda-Nakashima Atsushi Toyoshima Xiaojie Yin Hiromitsu Haba Koichi Fukase

Background/Objectives: Peritoneal dissemination of malignancies leads to poor prognoses, and no effective treatment currently exists. The difficulty of treating such malignancies is likely because systemically administered drugs cannot easily target malignant cells in the abdominal cavity. High intraperitoneal drug retention, non-toxicity towards normal tissues, and successful targeting of malignant cells are important for an effective therapy. The aim of this study was to evaluate an intraperitoneally administered astatine-labeled, integrin-targeted nanodrug, mPEG(Mn:350)-S-AuNP[211At]-c[RGDfK(C)] ([211At]AuNP@PEG/RGD), with respect to its kinetics, therapeutic efficacy, and safety. Methods: C6 rat glioma cells (107), and BxPC3 (107) and PANC-1 (107) human pancreatic cancer cells were seeded intraperitoneally into nude mice, and [211At]AuNP@PEG/RGD (0.979 ± 0.194 MBq for C6 models (n = 3), 1.139 ± 0.035 MBq for BxPC3 models (n = 10), and 1.308 ± 0.039 MBq for PANC-1 models (n = 10) per mouse) or saline were intraperitoneally administered 4–7 days later. Cytotoxicity against malignant cells, pharmacokinetics after administration, therapeutic efficacy, and safety in abdominal organs were evaluated. Results: Intraperitoneally administered [211At]AuNP@PEG/RGD accumulated exclusively in the peritoneal cavity for a long period of time and showed minimal systemic diffusion through the blood. In the C6 model, the intraperitoneal tumor mass was significantly lower in the treated group compared with that of the controls (p = 0.05). For the BxPC3 (median survival time: control/treated = 41/65 days, p < 0.001) and PANC-1 (median survival time: control/treated = 19/35 days, p < 0.001) peritoneal dissemination models, survival analysis revealed that [211At]AuNP@PEG/RGD significantly prolonged overall survival. Although transient weight loss, leukopenia, and thrombocytopenia were observed at one week post-administration, a short recovery trend was evident thereafter. One month after administration, no abnormalities were found in hematological tests or histological analyses of intra-abdominal organs. Conclusions: The intraperitoneal administration of astatine-labeled integrin-targeted [211At]AuNP@PEG/RGD nanoparticles showed promising findings in terms of safety and efficacy for treating peritoneally disseminated malignant tumors.

Pharmaceutics doi: 10.3390/pharmaceutics18091101

Authors: Kaiyan Su Yan Li Dongmei Zhang Yuxuan Zhou Jianping Zhang Hongyan Zhu Yun Zhao Cheng Guo Quanjun Yang

Background: Breast cancer remains the most common malignant tumor among women worldwide. Although conventional treatments including surgery, chemotherapy, and radiotherapy are effective, they are confronted with challenges such as tumor heterogeneity, systemic toxicity, and recurrence driven by drug resistance. To overcome these limitations, natural products have emerged as promising therapeutic alternatives owing to their multi-target efficacy and favorable biocompatibility. However, their clinical translation is still hindered by poor chemical stability, low aqueous solubility, and inadequate bioavailability. Nanodelivery systems offer a transformative solution by enhancing bioavailability and enabling precision targeting through the enhanced permeation and retention effect, thereby widening the therapeutic window and minimizing off-target toxicity. Purpose: This review evaluates diverse nanoparticle-based delivery systems and their targeting mechanisms in natural product-based breast cancer therapy. By examining inherent advantages and translational challenges, this analysis provides critical insights into the clinical development and application of these nanoformulations. Methods: A systematic literature search was performed in PubMed, ScienceDirect, Springer, Taylor & Francis, and Web of Science to identify relevant studies on natural product-based nanoformulations for breast cancer therapy. Results: Natural products exert anti-breast cancer effects through mechanisms such as inducing apoptosis, arresting the cell cycle, inhibiting invasion and metastasis, suppressing angiogenesis, and regulating autophagy. To overcome clinical hurdles, three complementary targeting strategies have been developed: passive, active, and stimuli-responsive targeting. These advances are shifting nanomedicines toward active precision therapy, markedly improving the therapeutic index. With multiple formulations already approved or in clinical pipelines, this field is rapidly progressing from laboratory research to clinical implementation. Conclusions: Natural products possess potent anti-breast cancer effects, and the application of nanodelivery technology effectively overcomes their inherent limitations of poor stability and low bioavailability. Although preliminary findings are promising, large-scale, randomized controlled clinical trials are urgently needed to systematically evaluate their safety, efficacy, and practical potential for clinical translation in breast cancer management.

Pharmaceutics doi: 10.3390/pharmaceutics18091100

Authors: Abhishek Dadhich Vikas Sharma Shivika Sharma Sweta Bawari Iyyakkannu Sivanesan

Type 2 diabetes mellitus (T2DM) remains a global health crisis, with nearly 589 million adults currently affected and projections pointing toward 853 million by 2050. Despite an expanding pharmacological armamentarium, a significant proportion of patients fail to achieve adequate glycaemic control, and the limitations of existing therapies, including adverse effects, cost, and limited accessibility, underscore the compelling need for novel therapeutic approaches. Phytochemicals such as curcumin, berberine, quercetin, resveratrol, and epigallocatechin gallate possess well-documented antidiabetic activity, operating through the PI3K/Akt, AMPK (activated protein kinase), NF-κB/JNK (nuclear factor kappa-B), and GLP-1R (glucagon-like peptide-1) signalling axes to improve insulin sensitivity, suppress gluconeogenesis, protect pancreatic beta-cells, and attenuate chronic metabolic inflammation. However, their clinical utility has been fundamentally constrained by poor oral bioavailability arising from low aqueous solubility, gastrointestinal instability, extensive first-pass metabolism, and P-glycoprotein-mediated efflux. Advanced drug delivery systems, including liposomes, solid lipid nanoparticles (SLN), nanostructured lipid carriers, PLGA (Poly (lactic-co-glycolic acid)) and chitosan nanoparticles, nanoemulsions, self-nanoemulsifying drug delivery systems, and phytosomes have demonstrated the capacity to overcome these barriers, achieving five- to ten-fold improvements in systemic bioavailability and substantially enhanced antidiabetic efficacy in preclinical models. Emerging mechanistic evidence further positions gut microbiota modulation and epigenetic reprogramming as additional therapeutic axes through which nano-encapsulated phytochemicals may exert durable metabolic benefits. Nonetheless, critical translational challenges persist, encompassing nanotoxicological risks, herb–drug interactions, the absence of harmonised regulatory frameworks for nano-phytomedicine products, phytochemical raw material variability, and formidable technical and economic barriers to scalable nanoparticle manufacturing. This review synthesises the current mechanistic, formulation, and clinical evidence within a unified analytical framework and identifies the strategic priorities of rigorous clinical development, regulatory clarity, and manufacturing standardisation required to translate nano-phytomedicine science into evidence-based T2DM therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18091099

Authors: Emily Cherny Tamara Minko

Background/Objectives: Intranasal delivery is a promising noninvasive approach to enhance central nervous system drug delivery in Parkinson’s disease (PD), potentially bypassing the blood–brain barrier and minimizing gastrointestinal side effects. This review summarizes clinical trials and translational evidence for intranasal PD therapies by treatment options and intent, highlighting key challenges in efficacy, pharmacokinetics, and safety. Methods: We reviewed human clinical trials, preclinical and pilot studies, and pharmacokinetic investigations of intranasal therapies for PD. Comparisons to established treatments were included in context. Only PD-specific clinical studies were analyzed. Therapies were grouped by rescue, antioxidant/metabolic/hormonal, and biologic or cell-based approaches, with continuous-delivery systems reviewed separately. Results: Intranasal rescue therapies, such as apomorphine, provide rapid improvement during OFF episodes (periods when the effects of PD medications fade and symptoms reappear), but tolerability issues and nasal irritation limit usage. Early-phase studies suggest intranasal delivery enables quick symptom relief and favorable pharmacokinetics, though most trials are small and focus on feasibility. New approaches include antioxidants, neurotrophic factors, gene therapy, and cell-based methods, with advanced formulations enhancing nasal retention and brain uptake. However, more translational evidence and long-term safety data are needed. Conclusions: Intranasal therapies for PD offer rapid rescue and expand options beyond standard drugs. Large, well-designed trials are needed to confirm efficacy, long-term safety, and optimal formulations. Intranasal delivery remains an emerging but potentially transformative strategy requiring further clinical research.

Pharmaceutics doi: 10.3390/pharmaceutics18091098

Authors: Shery Jacob Namitha Raichel Varkey Sai H. S. Boddu Jigar N. Shah Ibrahim Mustafa Abdi Rekha Rao

Extracellular vesicles (EVs) are naturally occurring nanoscale carriers that have gained attention as next-generation platforms for diagnostics, site-specific drug delivery, and tissue engineering owing to their high biocompatibility, minimal immunogenicity, and capacity to transport diverse bioactive cargo across biological barriers. This review discusses the classification, biogenesis, molecular constituents, and therapeutic properties of the major EV subtypes such as exosomes, microvesicles, and apoptotic bodies. It also highlights recent advances in EV engineering for cancer treatment, emphasizing immune modulation and targeted therapeutic delivery. Particular attention is given to plant-derived EVs, which have shown promise as scalable, low-toxicity nanotherapeutics with inherent bioactivity and effective drug delivery potential. Selected preclinical studies, recent patents, and ongoing clinical trials are also summarized, providing an up-to-date perspective on the clinical translation of EV-based technologies. Current challenges in EV isolation, characterization, scalable manufacturing, cargo loading, standardization, and regulatory approval, along with future directions for clinical translation, are summarized. Collectively, this review summarizes the growing applicability of EVs as next-generation platforms for precision medicine, targeted drug delivery, and regenerative therapies while identifying the major obstacles that must be addressed to facilitate their successful clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18091097

Authors: Adnan Amin Touseef Nawaz Oberdan Oliveira Ferreira Mozaniel Santana de Oliveira

Targeted protein degradation (TPD) represents a whole new paradigm in cell-level therapeutic design, with its ability to remove target proteins, normally through the endogenous proteasomal, lysosomal, or autophagic systems, rather than the traditional occupancy-driven inhibition approach. But the clinical efficacy of degraders is becoming more restricted based on delivery rather than efficacy only. Many proteolysis-targeting chimeras and new proximity-inducing systems have low solubility, are impermeable, are pharmacodynamically complicated, lack tissue selectivity, and cannot fully access the intracellular space. Nanomedicine and PD platforms could provide strategies not only to overcome these challenges, but also to provide other advantages, including enhancing exposure to degraders, biodistribution, controlled release, and context-dependent activation. This critical review is an outline of all lipid, polymeric, inorganic, biomimetic, targeted, activatable, and self-assembling delivery systems for TPD. We assess compositional considerations, in vitro and in vivo evidence, challenges for translation, and clinical endpoints required to support delivery-enabled degradation. Trusted TPD therapeutics need to relate different aspects of their design, such as degrader chemistry, carrier structure, disease biology, and pharmacodynamic biomarkers, to one another. Further investigations are needed to establish intact delivery of the degrader to the target, target depletion in relevant tissues, prolonged pharmacodynamics, favorable safety, and compelling therapeutic benefit relative to free degraders or traditional inhibitors. Thus, it is important to view delivery not simply as an additional step during formulation but as a design principle necessary for the reliable clinical outcome of degradation medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18091096

Authors: Ludwika Piwowarczyk Jagoda Szkudlarek Dariusz T. Mlynarczyk Szymon Tomczak Violetta Krajka-Kuźniak Aleksandra Majchrzak-Celińska Robert Kleszcz Ewelina Musielak Mateusz de Mezer Emilia Cicha Gabriela Anglart Anna Jelińska

Background/Objectives: Psoriasis is an incurable chronic immune-mediated inflammatory skin disorder for which effective topical treatment remains essential. Given the reported anti-inflammatory and dermatological potential of 20-hydroxyecdysone (20HE), this study evaluated its biological activity in normal and psoriatic keratinocytes and developed stable 20HE-loaded niosomes intended for potential future dermal delivery. Methods: The biological activity of 20HE was assessed in human epidermal keratinocytes (HEKs) and psoriasis-patient-derived keratinocytes (PHEKs) after 48 h of treatment by quantifying selected cytokines. Niosomes were prepared by thin-film hydration, rehydrated with water or citrate buffer, and sonicated. The formulations were characterized for particle size, polydispersity index, zeta potential, pH, encapsulation efficiency (EE), and 21-day storage stability. Potential in vivo toxicity was assessed in Danio rerio embryos by monitoring mortality, sublethal developmental changes, and locomotor behavior after exposure to nanoformulations for up to 96 h post-fertilization. Results: 20HE showed selective anti-inflammatory activity in PHEK cells, reducing IL-4, IL-17A, and IL-22 levels by 20–34% under elevated inflammatory conditions, without affecting healthy HEK cells. The formulations had particle sizes below 300 nm and acceptable polydispersity and zeta potential values, and they remained stable during storage. In the Danio rerio embryo model, the 1A/water caused sublethal developmental abnormalities at 96 hpf, whereas 1C/water (with 20HE) was associated only with mild pericardial edema after prolonged exposure. No significant behavioral alterations were observed across the tested concentration range, and the Lowest Observed Effect Concentration for 20HE was 20 μM. Conclusions: Free 20HE reduced the levels of IL-4, IL-17A, and IL-22 in PHEK cells, indicating its modulatory effect on cytokine production under the investigated inflammatory conditions. The developed 20HE-loaded niosomes showed favorable physicochemical properties and short-term stability, supporting their potential as candidate nanocarrier systems for future dermal application studies. The findings of the present study provide a basis for further investigation, particularly studies on drug release from the niosomal formulation and skin permeation from the final pharmaceutical dosage form.

Pharmaceutics doi: 10.3390/pharmaceutics18091095

Authors: Sara Hasan Seyedeh Negin Kassaee Derek J. Richard Nazrul Islam Emad L. Izake

Background/Objectives: The recent advances in pulmonary delivery have shifted the paradigm to the development of inhalable drug-loaded polysaccharide particles that can be positioned at the respiratory interface while reducing the systemic exposure. Quercetin has broad anticancer activity but remains difficult to translate because of its poor aqueous solubility, limited bioavailability and rapid metabolic loss. Here, we report quercetin-loaded chitosan/Salvia hispanica polysaccharide nanoparticles as a natural polyelectrolyte nanocarrier for pulmonary delivery in lung cancer. Methods: Central composite design (CCD) and artificial neural network (ANN) models were used for mapping the factors to responses. Results: The models displayed a higher predictive accuracy and optimization reliability for identifying the optimized formulation. The optimized nanoparticles showed a quasi-spherical morphology (mean hydrodynamic diameter = 331 ± 14.34 nm) with cationic ζ-potential of +36.3 ± 2.56 mV and polydispersity index of 0.15. The optimized formulation showed acceptable powder-flow characteristics, an encapsulation efficiency of 73.8 ± 0.73%, drug loading of 14.20 ± 0.22%, and biphasic release with sustained quercetin release over 48 h. In A549 and H460 cell lines, nanoencapsulation increased the antiproliferative effect of quercetin relative to the free compound, yielding lower IC50 values after 48 h of exposure. The nanoparticles showed greater suppression of wound closure, increased reactive oxygen species fluorescence and clear cellular uptake. Blank nanoparticles produced only limited effects. Conclusions: These results indicate that CCD/ANN-guided chitosan/Salvia hispanica polysaccharide nanoparticles provide a promising nanodelivery tool for quercetin delivery and enhanced in vitro activity in lung cancer cells, while the observed aerosol performance advocates further investigation of their pulmonary delivery potential.

Pharmaceutics doi: 10.3390/pharmaceutics18091094

Authors: Yongfeng Yang Tingting Song Kaili Huang Hong Huang Maoyuan Zhao Yi Li

Gene therapy and genome editing increasingly depend on the safe, effective, and cell-selective delivery of nucleic acids and protein–nucleic acid complexes. Although viral vectors remain important for applications requiring durable gene expression, non-viral vectors offer advantages in cargo capacity, modularity, transient expression, potential repeat dosing, and avoidance of vector–genome integration. Lipid nanoparticles (LNPs), polymeric nanoparticles, inorganic nanomaterials, extracellular vesicles (EVs), and biomimetic hybrid systems have consequently become central platforms for delivery of siRNA, mRNA, plasmid DNA, antisense oligonucleotides, and CRISPR-based genome editors. Among these, ionizable LNPs are currently the most clinically mature non-viral technology, supported by the clinical success of siRNA therapeutics and mRNA vaccines, as well as the emergence of in vivo CRISPR therapies. Nevertheless, efficient endosomal escape, cell-type-selective targeting, extrahepatic delivery, and repeat-dose tolerability remain substantial barriers. Polymeric vectors provide broad chemical tunability, allowing adjustment of charge density, degradability, stimulus responsiveness, intracellular trafficking, and cargo release. However, toxicity and batch-to-batch reproducibility remain key concerns. EVs provide a biologically derived alternative with favorable membrane interfaces and potential advantages for protein and ribonucleoprotein delivery, but their clinical translation is constrained by heterogeneity, loading efficiency, product characterization, and scalable manufacturing. This review summarizes recent advances in non-viral gene-delivery platforms, compares their strengths and limitations, and discusses future directions in cell-selective delivery, endosomal escape, transient delivery of genome-editing machinery, engineered EVs, hybrid vectors, and manufacturing-oriented development. The field is transitioning from organ-level delivery toward delivery of the correct payload to the correct cell type at a clinically relevant exposure and safety margin.

Pharmaceutics doi: 10.3390/pharmaceutics18091093

Authors: Jahanvi Patel Michael Stolinski Anil Vangala

Insulin delivery via the buccal route has shown long-standing promise for diabetes therapy, but clinical translation remains limited. Most systems address only part of the delivery problem, whereas a viable product must simultaneously provide adequate insulin residence time, effective mucus and epithelial permeation, stability, dose consistency, safety, and manufacturability. This review examines the biological and formulation barriers limiting buccal insulin delivery and compares major platform types, including mucoadhesive films and patches, nanocarrier-based systems, deformable vesicles, chemistry-led permeation approaches, and device-enabled strategies. Stability, alongside limited permeability, is considered a key barrier, while aggregation and excipient-related instability remain insufficiently addressed. This review also discusses why promising preclinical findings often fail to translate, exhibiting low bioavailability, variability, safety concerns, and scale-up challenges recurring across platforms. Overall, further progress in buccal insulin delivery will depend on formulation strategies that better integrate permeation enhancement, stability preservation, and translational feasibility.

Pharmaceutics doi: 10.3390/pharmaceutics18091092

Authors: Anna Simonsson Nicklas Bunta Sundin Tobias Bramer Alex Wimbush Göran Alderborn

Background/Objectives: Packing, flow, and aerosolization properties of a series of binary adhesive mixtures containing micronized or spray-dried drugs were investigated, and the relationships between these blend properties and the blend structure were studied. Methods: Micronized or spray-dried terbutaline sulfate and salbutamol sulfate were used as model drugs, and an α-lactose powder was used as the carrier. Binary mixtures with drug loads ranging from 2 to 20% were prepared. The bulk density, compressibility, permeability, and shearing properties of the carrier powder and mixtures were determined, along with the in vitro aerosolization propensity of the mixtures using two types of inhalers. Imaging of the mixtures was used to assess the blend structure. Conclusions: The particle engineering method gave differences in particle crystallinity and morphology. The development of the adhesive layer with drug load was broadly consistent with the blend state concept. Spray-dried particles, however, exhibited a higher propensity to localize within surface cavities on the carrier and produced a more voluminous enveloped adhesive layer. The spray-dried particles gave a higher bulk density, a lower Hausner ratio, comparable shear strength, and a lower angle of internal friction. Aerosolization performance, including metrics such as fine particle fraction (FPF), depended on inhaler design; nevertheless, for both inhalers, aerosolization behavior was influenced by blend state and physicochemical properties of the drug. At low drug loads, spray-dried particles dispersed to a lower degree, while at high drug loads, the dispersion performance of the two particle types converged. For example, at an intermediate drug load of 7.4%, the FPF was about 20% for the spray-dried drugs and about 30% for the micronized drugs using the Screenhaler device, while the corresponding FPF:s were about 25% and 50% for spray-dried drugs and 40% and 55% for crystalline drugs using the Monodose inhaler. Overall, the physical characteristics of the drug particles were found to influence the structural evolution of the blends, as well as their mechanical and aerosolization properties.

Pharmaceutics doi: 10.3390/pharmaceutics18091091

Authors: Lania Ali Hussein Alexandra Nedic Andrejs Sitovs Valentyn Mohylyuk

Although oral peptide drug administration is the most desirable for both patients and healthcare systems, it is a major pharmaceutical challenge. Typically, less than 1% of orally administered peptide therapeutics reaches the systemic circulation. Modern research has focused on overcoming the physiological barriers responsible for the poor peptide drug bioavailability, while little attention has been given to understanding how measurable absorption occurs despite the barriers. This review aimed to explore the physiological factors that influence oral peptide absorption, such as enzymatic degradation and pH effects, motility, microbiota, transporters, tight junctions, and the effect of inflammation. The available data indicate that these factors function not only as barriers but may also create limited opportunities for peptide uptake. Enzymatic degradation and microbial metabolism reduce the availability of intact peptide drugs yet may generate biologically relevant fragments. Tight junctions are generally regarded as restrictive barriers but are shown to exhibit dynamic regulations that may permit transient paracellular transport. Similarly, gastrointestinal motility, microbiota, and inflammatory processes influence epithelial permeability and duration of peptide exposure both to absorptive surfaces and to degradative enzymes. In contrast, transporter-mediated uptake appears unlikely to account for the absorption of intact peptide drugs due to their substrate size limitations. Together, the findings suggest that measurable oral peptide absorption is unlikely to result from a single dominant pathway but rather from the cumulative contribution of the mechanisms. Improved understanding of these endogenous mechanisms may open opportunities to enhance oral peptide bioavailability and support the development of more effective therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18091090

Authors: Chao Fang Lixin Xie Daihan Xie Jingting Yin Shiji Zhang Yiling Gan Xiaodong Hou Fanlei Kong Yu Huo Xiuli Liu

Introduction: Acute lung injury (ALI) is characterized by severe inflammation and oxidative stress. However, effective pharmacological interventions for ALI remain limited. Carnosine, an endogenous dipeptide known for its redox-regulating and immunomodulatory activities, has demonstrated promising protective effects. In the present study, inhalable carnosine-loaded liposomes (Cn-Ls) were developed to enhance pulmonary delivery and achieve localized treatment in a lipopolysaccharide (LPS)-induced ALI model in mice. Methods: Cn-Ls were prepared and systematically evaluated for their morphology, stability, drug-loading capacity, and release kinetics. In vitro assays were performed to evaluate their cytocompatibility, antioxidant activity, and effects on LPS-induced reactive oxygen species (ROS) generation. In an ALI model, inhaled Cn-Ls were administered to assess pulmonary retention and therapeutic efficacy, including lung inflammation, oxidative stress, circulating levels of C-reactive protein (CRP), tumor necrosis factor (TNF)-α, interleukin (IL)-6, lung architecture, and respiratory function. Results: Encapsulation of carnosine within liposomes markedly prolonged its pulmonary retention (t1/2 = 1.7 h vs. 1.0 h for free carnosine), providing a more sustained lung-retentive delivery profile for up to 12 h. In vitro assays showed that Cn-Ls had excellent cytocompatibility, reduced cell death, exhibited potent antioxidant activity, and effectively suppressed LPS-induced ROS generation. In an ALI model, inhaled Cn-Ls markedly mitigated lung inflammation and oxidative stress, reduced circulating levels of CRP, TNF-α, and IL-6, preserved lung architecture, and improved respiratory function. Compared with free carnosine, Cn-Ls exhibited enhanced pulmonary retention and superior therapeutic efficacy. Conclusions: These results identified inhalable Cn-Ls as a potential nanotherapeutic approach for targeted ALI management and provided a foundation for further translational development. However, additional investigations are required to assess the long-term safety of Cn-Ls, optimize formulation stability and scalability, and further elucidate the underlying therapeutic mechanisms before clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18091089

Authors: Smita Salunke Chi Kin Anthony Chan Sifan Hu

Background/Objectives: The administration of medicines via enteral feeding tubes presents unique challenges, including dose loss, tube blockage, and user-dependent variability. Although Quality by Design (QbD) frameworks provide a structured approach to product quality, administration-related quality attributes are rarely assessed in a systematic manner during pharmaceutical development. A conceptual framework, termed the Quality Target Administration Profile (QTAP), was developed to extend QbD principles to the administration stage. Methods: The framework comprises three phases: administration context characterization, administration attribute identification, and risk-based determination of Critical Administration Attributes (CAAs). QTAP was applied to three representative EFT administration scenarios: a ready-to-use oral suspension, a crushed tablet, and an oral solution. Results: Application of the framework identified 18–32 administration attributes across the three representative scenarios and prioritized 5–6 CAAs per scenario according to clinical impact, variability risk, and human-factor considerations. Administration quality requirements varied across scenarios, reflecting differences in administration context, including dosage form, patient population, clinical setting, and user characteristics. QTAP provides a structured approach for identifying and prioritizing administration-related quality considerations within pharmaceutical development. Conclusions: As a conceptual framework, QTAP is intended to complement rather than replace existing QbD approaches. Future prospective validation, including human-factor studies and real-world evaluations, is needed to establish its practical utility and predictive value.

Pharmaceutics doi: 10.3390/pharmaceutics18091088

Authors: Hao-Yuan Lee Yu-Chia Chang Chyi-Liang Chen Shu-Hua Ko Yu-Ling Huang Shang-Po Shen Yu-Lung Hsu Wen-Yuan Lee Hung-Chih Lin

Background/Objectives: Autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD) substantially affect quality of life. Previous studies have not evaluated whether the associations between early-life antibiotic exposure and the risks of ASD and ADHD differ according to the antibiotic agents administered during infancy. Methods: This nationwide case–control study used Taiwan’s National Health Insurance Research Database, Birth Reporting Database, and Maternal and Child Health Database. The cohort included 1,693,718 term neonates born between 2004 and 2015 and followed through to 2022. For ASD (n = 7265) and ADHD (n = 39,056) cases, antibiotic-exposed children were matched 1:1 with unexposed children based on sex, gestational age, birth weight, and birth year. Adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) were estimated using multivariable Cox proportional hazards models. Results: Oral antibiotic exposure was associated with lower ASD but higher ADHD risk, while parenteral exposure was associated with increased risks of both outcomes. Amoxicillin and erythromycin exposure during the first year of life were associated with lower ASD risk, whereas ampicillin exposure at 4–12 months was associated with higher ASD risk (all p ≤ 0.033). For ADHD, exposure to amoxicillin/clavulanic acid, cefixime, ampicillin, and ampicillin/sulbactam was associated with increased risk, whereas erythromycin, cephalexin, and sulfamethoxazole-trimethoprim were associated with reduced risk (all p < 0.047). Conclusions: Associations between early-life antibiotic exposure and the risks of ASD and ADHD varied according to antibiotic agents, route of administration, and timing of exposure. These findings may help explain inconsistencies among previous studies and inform antibiotic prescribing during infancy.

Pharmaceutics doi: 10.3390/pharmaceutics18091084

Authors: Luigi Ruccolo Aleksandra Evangelista Francesco Andresini Rossella Dorati Ida Genta Marco Benazzo Pietro Canzi Elena Carlotto Bice Conti Silvia Pisani

Background/Objectives: Shape-memory biodegradable scaffolds (4D scaffolds) represent promising platforms for minimally invasive tissue engineering and localized drug delivery. This study investigated how two different fabrication techniques, electrospinning (ES) and extrusion-based direct ink writing (DIW), influence the structural, thermal, mechanical, shape-memory, and drug-release properties of poly(L-lactide-co-caprolactone) (PLA/PCL 70:30) scaffolds loaded with dexamethasone (DXM). Methods: DXM-loaded PLA/PCL 70:30 scaffolds were fabricated by ES and DIW. The resulting matrices were characterized in terms of morphology, mass, thickness, drug-loading efficiency, thermal properties by differential scanning calorimetry, shape-memory performance, tensile mechanical properties, and in vitro DXM release. Results: Both fabrication techniques produced DXM-loaded matrices with comparable mass and thickness and high loading efficiencies (>82%). Glass transition temperatures ranged between 33 and 39 °C, supporting thermally induced shape recovery under physiologically relevant conditions, while ES processing was associated with higher polymer crystallinity. All scaffolds exhibited shape-memory behavior, with recovery ratios exceeding 90%. ES scaffolds displayed a microporous nanofibrous architecture, whereas DIW scaffolds showed a more open and highly porous structure. These morphological differences were reflected in their mechanical behavior: ES scaffolds exhibited higher tensile strength (up to 16.5 MPa vs. 1.9 MPa) and elongation at break (up to 320% vs. 243%). Drug-release profiles were also fabrication-dependent, with ES scaffolds reaching a plateau at approximately 80% DXM release, whereas DIW scaffolds showed near-complete release within 48 h. Conclusions: Both fabrication approaches preserved the thermoresponsive shape-memory behavior of PLA/PCL 70:30 but generated distinct scaffold architectures that strongly influenced mechanical performance and DXM-release kinetics.

Pharmaceutics doi: 10.3390/pharmaceutics18091087

Authors: Pierpaolo Palermo Davide De Angelis Elisa Sgarbi Irene Bassanetti Michael M. Tunney Dimitrios A. Lamprou

Backgorund/Objectives: Respiratory diseases represent a substantial global health burden and require effective localised pulmonary delivery strategies, particularly for poorly water-soluble therapeutic molecules. Nanoparticle-based drug delivery systems, especially those manufactured using microfluidics, have emerged as promising approaches to overcome pulmonary barriers, enhance local drug retention, and reduce systemic side effects. Among these nanocarriers, solid lipid nanoparticles (SLNs) and solid hybrid nanoparticles (SHNs) combine biocompatibility with controlled release and improved formulation stability. Methods: In this study, SLNs and lipid–chitosan SHNs were developed using microfluidic technology as candidate platforms for pulmonary drug delivery, with Cyclosporine A (CyA) used as a model hydrophobic cyclic peptide. Nanocarriers were produced using 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol as lipids, with low-molecular-weight chitosan incorporated to obtain hybrid systems. Physicochemical properties were evaluated using dynamic light scattering (DLS) and ζ potential measurements, while morphology and structural organisation were investigated using transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). Results: The microfluidic approach enabled the production of nanoparticles with controlled sizes below 200 nm, narrow size distributions, and good reproducibility. In addition, the SHNs exhibited a positive surface charge, high encapsulation efficiency (~80%), and good colloidal and thermal stability. In vitro release studies showed an initial burst release followed by sustained CyA release, reaching approximately 94% cumulative release within 6 h. The Korsmeyer–Peppas model was used as the standard kinetic model. No blank nanoparticles were used as controls in the EE and release assay. Conclusions: Overall, these findings support further investigation of microfluidic-produced lipid and hybrid nanoparticles as candidate platforms for pulmonary drug delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18091086

Authors: Jessica R. Boulos Karen E. Russi Jordan Kinnitt Daphne Gomez Escudero Tyler Willis Jorrian Abadeer Emalee Mann Aaron Cristina Anderson Angela Alexander-Bryant

Background/Objectives: RNA interference (RNAi) represents a promising therapeutic approach for silencing oncogenes involved in cancer progression by utilizing small interfering RNA (siRNA). However, siRNA requires an efficient delivery system to overcome cellular uptake and endosomal escape barriers. This study aimed to evaluate a multifunctional tandem peptide, GE11-599, designed to enhance the targeted delivery of siRNA and maintain its bioactivity in glioblastoma (GBM) cells. Methods: The GE11-599 peptide, consisting of an EGFR-targeting GE11 motif and a 599 fusogenic domain, was complexed with siRNA via electrostatic interactions to form nanoparticles. We assessed nanoparticle physicochemical properties, protection of siRNA from serum and RNase degradation, and cellular uptake in two GBM cell lines (U118MG and U87MG). Mechanistic studies evaluated receptor-mediated endocytosis and the subsequent escape from endosomes. Functional assays quantified STAT3 gene silencing and downstream effects on cell migration following treatment with GE11-599–siSTAT3 complexes. Results: GE11-599 formed positively charged, monodisperse nanoparticles capable of protecting siRNA from degradation. The tandem peptide significantly enhanced cellular internalization through EGFR-mediated endocytosis and facilitated endosomal escape of siRNA. Treatment with GE11-599–siSTAT3 resulted in robust gene silencing, achieving up to an 80% reduction in STAT3 mRNA expression. Downstream functional assessment showed a 40% decrease in migration in GBM cells treated with GE11-599–siSTAT3 complexes. Conclusions: The GE11-599 tandem peptide effectively enhances cell-specific internalization and endosomal escape of siRNA in GBM cells, resulting in increased siRNA bioactivity and functional gene silencing. These findings support GE11-599 as a promising siRNA delivery platform for targeting EGFR-expressing cancers.

Pharmaceutics doi: 10.3390/pharmaceutics18091085

Authors: Yitong Yang Dandan Guo Wei Chen Binbin Sun Mengyu Qiu Kai Wang Wenbo Dou Kang Wang Zhanjiang Zhang Shuying Feng

Background: Methotrexate (MTX) is a well-established therapeutic agent for psoriasis owing to its anti-inflammatory and immunomodulatory effects. However, its clinical use is limited by insufficient local accumulation in skin lesions and the potential risk of systemic exposure. Dunaliella salina-derived exosome (DsEXO) has favorable biocompatibility, low immunogenicity and potential skin delivery capacity, making it a promising natural nanocarrier for topical MTX delivery. This study aimed to construct Dunaliella salina-derived exosome loaded with methotrexate (DsEXO@MTX) and evaluate its therapeutic efficacy and potential mechanisms in psoriasis-like skin inflammation. Methods: DsEXO@MTX was prepared and characterized in terms of morphology, particle size, surface charge and drug-loading capacity. Cellular uptake, skin retention and tissue distribution were evaluated using fluorescence imaging and skin section analysis. Therapeutic efficacy was evaluated in an imiquimod-induced psoriasis-like mouse model by clinical scoring, histopathological examination, spleen index measurement and Ki-67 immunofluorescence staining. STAT3 phosphorylation and Th17/Treg differentiation were further examined to explore the potential immunomodulatory mechanism. Results: DsEXO@MTX exhibited a relatively uniform particle size distribution and drug-loading capacity. In vivo fluorescence imaging and skin section analysis showed that DsEXO@MTX enhanced local skin retention and promoted fluorescence distribution in epidermal and dermal regions. It significantly alleviated IMQ-induced erythema, scaling, epidermal thickening, inflammatory infiltration, splenomegaly and abnormal keratinocyte proliferation in psoriasis-like mice. Mechanistically, DsEXO@MTX reduced STAT3 phosphorylation and modulated Th17/Treg differentiation, suggesting restoration of immune balance in psoriatic inflammation. Conclusions: DsEXO@MTX represents a natural exosome-like nanovesicle-mediated topical MTX delivery system that improves local drug delivery and enhances therapeutic efficacy in IMQ-induced psoriasis-like skin inflammation, particularly when administered topically. These findings provide a potential strategy for safer and more efficient local treatment of psoriasis.

Pharmaceutics doi: 10.3390/pharmaceutics18091083

Authors: Adnan Murad Bhayo Ying Li Alaa R. Aboushanab Junyi Lin Ashkan Hassankhanirad Wei Li Jingjing Sun

Triple-negative breast cancer (TNBC) is an aggressive and heterogeneous subtype lacking effective targeted therapies. Its tumor microenvironment (TME) exhibits distinct features, including acidity, redox imbalance, elevated reactive oxygen species (ROS), and hypoxia, which provide exploitable triggers for targeted drug delivery. Stimuli-responsive polymeric nanocarriers have emerged as promising platforms that enable spatiotemporally controlled and site-specific therapeutic release in response to these endogenous cues, as well as exogenous stimuli such as temperature and light. These systems improve drug accumulation, penetration, and therapeutic efficacy while reducing systemic toxicity. Unlike previous reviews that broadly discuss nanocarriers in cancer therapy, this review focuses on the structure–function relationships of TME-responsive polymeric systems in TNBC and their translational limitations. We summarize recent advances in pH-, redox-, ROS-, hypoxia-, photo- and temperature-responsive polymers, highlighting their design strategies and therapeutic applications. Key challenges, including stimulus heterogeneity, limited in vivo validation, and clinical translation barriers, are also discussed. This review provides a concise framework for the rational design of programmable, multi-responsive polymeric nanomedicines for TNBC therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091082

Authors: Abdullah A. Assiri

Oral targeted therapies now constitute a substantial and growing proportion of anticancer drug therapy, shifting administration from the controlled intravenous setting to patient-managed oral therapy in the outpatient setting, where systemic exposure depends on factors that parenteral therapy largely bypasses. Food effects, gastric pH, first-pass metabolism, transporter activity, organ function, concomitant medications and adherence contribute to variability in exposure, and many oral anticancer agents have narrow therapeutic indices in which modest exposure changes carry clinical consequence. This review synthesizes the pharmacokinetic determinants of oral anticancer drug exposure across twenty-seven exemplar agents spanning the main mechanistic classes and translates them into actionable pharmacy practice. Its contribution is a cross-class agent-level comparison in which within-class divergences are made explicit, the integration of determinants usually reviewed separately, and an explicit statement of the evidence level behind every entry. We examine food effects, the interaction between acid-suppressive therapy and pH-dependent agents, the dominant role of cytochrome P450 3A4 and of the efflux transporters P-glycoprotein and breast cancer resistance protein, and the exposure–response relationships that motivate therapeutic drug monitoring, for which the evidence remains uneven and does not yet support routine use. We propose a structured framework for operationalizing these principles in daily practice.

Pharmaceutics doi: 10.3390/pharmaceutics18091081

Authors: Na-Young Yu Kyu-Jin Cho Saeeun Ryu Gyulim Kim Jae-Woo Shin Donghee Park Jongho Won Young-Kee Shin Eun-Ah Kim Sang-Goo Cho Nae-Won Kang Byung-Hoon Lee Nam-Young Kim Eun-Seong Kim Dae-Duk Kim

Background/Objectives: Androgenetic alopecia (AGA) is the most prevalent form of hair loss. Although topical minoxidil (MNX) is widely used to treat AGA, its efficacy is limited by poor penetration across the stratum corneum. This study evaluated radiofrequency (RF) microporation as a means of enhancing cutaneous MNX delivery and hair-regrowth efficacy in a dihydrotestosterone (DHT)-induced AGA mouse model. Methods: RF-induced skin permeabilization and barrier recovery were assessed in rats using methylene blue and rhodamine B staining. In vivo skin deposition and pharmacokinetic studies were conducted to quantify cutaneous MNX accumulation and systemic exposure. Hair-regrowth efficacy was evaluated in DHT-treated mice. Results: RF microporation generated transient microchannels in the stratum corneum, increased rhodamine B penetration into deeper skin layers, and allowed substantial barrier recovery within 24 h. RF pretreatment significantly increased MNX deposition in the epidermis/dermis by 2.40-fold at 1 h and 1.94-fold at 3 h compared with topical MNX alone. RF-assisted topical administration resulted in a relative bioavailability of 11.17%, compared with 4.74% for topical administration without RF, while dose-normalized systemic exposure remained substantially lower than that following oral administration. In the AGA model, RF-assisted MNX treatment significantly increased hair coverage, length, and shaft thickness. Histological analysis further showed more prominent follicular structures in the RF-assisted MNX groups. Conclusions: RF microporation creates transient microchannels that enhance cutaneous MNX delivery and improve hair-regrowth efficacy. Importantly, RF-assisted topical administration maintained substantially lower systemic exposure than oral administration, supporting its potential as a needle-free strategy for topical AGA therapy and further translational evaluation.

Pharmaceutics doi: 10.3390/pharmaceutics18091080

Authors: Justyna Srebro Witold Brniak Paulina Poloczek Marian Paluch Aleksander Mendyk

Background/Objectives: Bisoprolol fumarate (BF) is a cardioselective beta-blocker used to treat pediatric heart failure and certain tachyarrhythmias. Currently, it is administered mainly as extemporaneously prepared suspensions from crushed tablets, which may cause issues with dose accuracy, stability, and palatability. Orodispersible minitablets (MODTs) may facilitate administration and individualized dosing, while prolonged release could potentially reduce peak-to-trough fluctuations and, thus, minimize adverse effects such as hypotension or bradycardia. This proof-of-concept study aimed to develop spray-dried prolonged-release microparticles containing BF, incorporate them into MODTs, and evaluate the effect of compression on drug release. Methods: Microparticles were prepared by spray drying ethanolic solutions of BF and ethylcellulose at 40 °C, 50 °C, and 60 °C. Their characterization included scanning electron microscopy, X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis and dissolution studies. Selected microparticles were compressed into 3 mm MODTs, which were evaluated for mechanical properties, disintegration, and BF release. Results: Formulations containing 10% BF and 90% ethylcellulose released from 52.6% to 73% of BF after 2 h, increasing to 84.8–90.0% after 8 h and reaching 96% after 24 h. Solid-state analyses indicated complete amorphization of the drug in these microparticles. The spray-drying temperature affected process efficiency but did not significantly impacted morphology or dissolution behavior. The optimized MODTs disintegrated in less than 30 s and had a tensile strength of up to 2.62 MPa. Importantly, compression increased the initial BF release from 26.4% for the microparticles to 43.9% for the MODTs at 0.5 h, resulting in f2 values of 45.04–48.46. Despite the higher initial BF release, the moderately prolonged-release profile was maintained in the case of MODTs. Conclusions: These findings demonstrate the feasibility of combining prolonged-release microparticles with orodispersible minitablets as a proof-of-concept for a pediatric drug delivery approach.

Pharmaceutics doi: 10.3390/pharmaceutics18091079

Authors: Mark Mandrik Veronika Makarova Ludmila Korol Ivan Sadkovskii Ivan Krasnyuk Sergey Antonov

Background: Hot-melt extrusion (HME) is a scalable pharmaceutical technology increasingly relevant to flexible manufacturing, including small-batch production, personalized dosage-form development, and potential use in pharmacy compounding. When translated into compounding practice, however, HME introduces a risk that routine quality-control methods available in pharmacies may be insufficient to reliably assess the stability of extrusion-based preparations. Methods: Granules containing 50% (w/w) darunavir were prepared by HME at 70 and 90 °C using a previously developed polymeric premix. Samples were stored for 24 months under ambient conditions. During storage, routine quality attributes were evaluated, including appearance, particle size distribution, loss on drying, disintegration time, content uniformity, and assay. Solid-state changes were investigated using differential scanning calorimetry (DSC) and X-ray diffraction (XRD), with a reference PEG-associated darunavir sample prepared and characterized for comparative analysis. Changes in drug release and darunavir content were assessed by dissolution testing and HPLC analysis, respectively. Results: Granules produced at both extrusion temperatures retained acceptable routine quality attributes throughout the 24-month storage period. No substantial changes were detected by visual inspection, pharmacopoeial tests, or UV assay. However, DSC revealed a new thermal event after storage, while XRD showed the formation of a new crystalline phase. Comparison with the reference PEG-associated sample supported the assignment of this phase as a PEG-associated crystalline phase of darunavir. Importantly, this transformation occurred even though the routine quality attributes evaluated in pharmacy compounding practice remained unchanged. Dissolution profiles differed between samples tested immediately after preparation and after long-term storage, with a more pronounced overall difference for granules produced at 90 °C, whereas HPLC confirmed comparable darunavir content in all investigated samples. Discussion: Our results show that routine compounding quality control can meet conventional acceptance criteria while failing to detect API solid-state changes in the investigated HME-derived system. In the PEG-containing matrix, amorphous darunavir undergoes storage-induced crystallization, forming a PEG-associated crystalline phase consistent with its known affinity for polyol-containing media. Conclusions: Acceptable routine quality attributes do not necessarily reflect the solid-state stability of APIs in HME-based formulations. These results highlight the need for solid-state risk assessment when developing extrusion-based systems intended for pharmacy compounding and other personalized manufacturing models in which routine quality control may not include advanced solid-state characterization.

Pharmaceutics doi: 10.3390/pharmaceutics18091078

Authors: Mantosh Kumar Satapathy Ting-Lin Yen Jing-Shiun Jan Ruei-Dun Tang Jia-Yi Wang Rajeev Taliyan Chih-Hao Yang

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Pharmaceutics doi: 10.3390/pharmaceutics18091077

Authors: Diana-Maria Mateescu Dragos-Mihai Gavrilescu Ruxandra-Ioana Mincioaga Ana-Maria Pah Ana-Olivia Toma Daniela-Vasilica Serban Cristiana Adina Avram Maria-Laura Craciun Bogdan Enache Camelia-Oana Muresan

Background/Objectives: Glycyl-L-histidyl-L-lysine (GHK) and its copper(II) complex GHK-Cu have been studied for matrix remodeling, inflammation, redox regulation, angiogenesis, and tissue repair, yet the literature frequently treats GHK-Cu as a single active ingredient despite formulation-dependent variation in coordination state, speciation, stability, pharmacokinetics, and toxicity. We critically evaluate GHK-Cu simultaneously as a bioactive metallopeptide and as a drug-delivery cargo, with explicit separation of apo-GHK, canonical GHK-Cu, GHK-derived copper peptides, and non-GHK copper-peptide systems. Methods: We conducted a structured systematic evidence-mapping review of PubMed/MEDLINE, Europe PMC, major publisher platforms, ClinicalTrials.gov, backward citation chains, and official European Union, U.S., and ICH regulatory sources from database inception through 12 August 2026. Biological evidence level and chemical/formulation quality were graded independently using an author-defined two-axis framework. Delivery studies were extracted against a fixed matrix comprising formulation, claimed loading, molar copper occupancy, labile copper, species-resolved release, factorial controls, stability/manufacturability, and objective outcome. Quantitative pooling was not performed because active-entity definitions, formulations, doses, models, comparators, and endpoints were not quantitatively commensurable. Results: Preclinical data consistently support effects on matrix remodeling, epithelial repair, inflammatory/redox regulation, and angiogenesis, but the clinical evidence remains sparse and does not meet contemporary active-entity quality standards. Historical cosmetic reports are small or incompletely characterized; a 13-participant post-CO2-laser study was negative on objective endpoints, whereas a 2026 18-participant split-face eyebrow study reported positive cosmetic hair outcomes but did not define GHK-Cu speciation or local exposure. The ongoing phase 2 acute-wound study NCT07437586 remains recruiting and has no efficacy results; its registration cannot be used as evidence of clinical translation. Across delivery studies, particle size, polydispersity, encapsulation efficiency, total peptide/copper content, and bulk release are often reported, whereas molar occupancy, labile copper, and release of intact GHK-Cu versus apo-GHK/free copper are usually not resolved. Conclusions: Translation is limited less by biological plausibility than by pharmaceutical definition and evidence attribution. We define the “active pharmaceutical entity” operationally as the reproducible chemical state intended to mediate pharmacology at administration, not as an established regulatory designation or a claim that one immutable molecular species persists in biological fluids. We further propose, explicitly as an author-derived development framework rather than a consensus standard, a control strategy based on molar occupancy, route-specific labile copper specifications, orthogonal speciation, mechanism-linked potency, species-resolved release, factorial controls, route-specific safety decision thresholds, ICH-aligned stability, and GMP-scalable manufacture. Until these requirements are met and controlled clinical efficacy is demonstrated, GHK-Cu should be regarded as a promising but unproven therapeutic cargo rather than a clinically validated regenerative drug.

Pharmaceutics doi: 10.3390/pharmaceutics18091076

Authors: Agata Olszewska Maria Brycka Anita Umerska Lidia Tajber Marek Pyda Marcin Skotnicki

Objective: In this study, the solid-state properties of two amorphous forms of rosuvastatin calcium (ROS) and co-amorphous systems of rosuvastatin with carvedilol (CAR) were investigated. Methods: The studied systems were investigated using TGA, DSC, TMDSC, PXRD, SAXS, SSNMR, and DVS. Results: DSC, TMDSC and SAXS revealed subtle differences between as-received ROS (ROS AR) and freshly prepared amorphous ROS (ROS AM), providing evidence of relaxation polyamorphism. Analysis of the scanning-rate dependence of the glass transition temperature classified ROS as a fragile glass former. Water sorption studies of ROS AR using Brunauer–Emmett–Teller analysis, the Guggenheim–Anderson–de Boer equation, and Young–Nelson models indicated a moderate specific surface area and multilayer adsorption, with water binding both at the surface and within the bulk. Co-amorphous CAR–ROS systems in various molar ratios exhibited single glass transitions, confirming the formation of homogenous amorphous phases. Negative deviations from the Couchman–Karasz equation indicated non-ideal mixing behaviour. Physical ageing studies showed high stability of the ROS AM and CAR-ROS 1:1 co-amorphous system, which remained amorphous after three years of storage. Conclusions: Overall, ROS forms stable amorphous and co-amorphous systems with CAR, exhibiting favourable miscibility, structural stability, and potential for application in fixed-dose combination formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18091075

Authors: Nader Amanatchi Ilyesse Bihi Matthieu Briet Wim De Malsche Karine H. Hellemans

Background/Objectives: Poly(lactic-co-glycolic acid) (PLGA) microparticles are used for sustained drug delivery, yet their final quality depends on both initial droplet formation and the subsequent solvent-extraction-driven droplet-to-particle transition. Microfluidics provides excellent control over precursor droplets, but this control may be partially lost during particle formation. We previously developed dynamic solvent extraction (DSE), in which the droplet-to-particle transition occurs progressively during continuous transport through an extended microfluidic channel. Here, we investigated whether this control improves quality attributes and release of drug-loaded PLGA microparticles. Methods: Dexamethasone (DEX)-loaded PLGA microparticles were produced using DSE or static solvent extraction (SSE), in which microfluidic droplets were transferred to an external aqueous medium for particle formation. Three PLGA concentrations (2.5, 5, and 10% (w/v)) were investigated, with processing conditions selected to obtain comparable final particle sizes and reduce size as a confounding variable. Results: DSE significantly reduced particle-size coefficient of variation compared with SSE, indicating better preservation of size uniformity. Drug loading and encapsulation efficiency were governed mainly by PLGA concentration, with no significant effect of extraction strategy. Morphological effects were formulation-dependent, with the most pronounced defects in 5% (w/v) SSE particles. All formulations provided sustained DEX release over 70 days. Although the overall effect of extraction strategy on 24 h burst release was not statistically significant, DSE consistently produced numerically lower burst release, with the largest and most variable burst observed for the morphologically heterogeneous 5% (w/v) SSE formulation. Conclusions: These findings extend DSE to drug-loaded sustained-release microparticles and identify the solvent-extraction environment as an important determinant of final particle quality.

Pharmaceutics doi: 10.3390/pharmaceutics18091074

Authors: Maria Medina de Azevedo Maria Luiza Fidelis da Silva Gabriela Pereira da Silva Joyner David Anaya Miranda Annye Vitória Moraes Luana Ale Bertoncello Pael Telma Lélia Gonçalves Schultz de Carvalho Giselle Nathaly Calaça Thainá Aparecida Rafael Silva Bianca Viana Silva Thaylla Bianca de Almeida Vilela João Pedro Vilella Neto Ariany Carvalho dos Santos Ana Paula de Carlos Sela Monique Assis de Vasconcelos Barros Francislaine Aparecida dos Reis Lívero Otávio Akira Sakai Érica Marusa Pergo Coelho Arquimedes Gasparotto Junior

Background: Hypertensive disorders of pregnancy present a major global health challenge. Green nanotechnology offers a strategy to enhance the stability and efficacy of bioactive natural products. This study evaluated gold nanoparticles green-synthesized with Pfaffia glomerata extract (AuNPs-PG) against gestational hypertension and fetal complications in spontaneously hypertensive rats (SHRs). Methods: P. glomerata root extract was characterized by UHPLC-MS/MS. AuNPs-PG were green-synthesized with 2 mmol/L HAuCl4 (1:5 v/v) at 40 °C and characterized by UV-Vis and DLS. Pregnant Wistar-Kyoto and SHRs were divided into naive, negative control (NC), amlodipine (5 mg/kg), and AuNPs-PG groups (0.03, 0.1, and 0.3 mg/kg) treated daily on gestational days 1–18 to evaluate maternal cardiovascular, renal, biochemical, and reproductive outcomes. Results: UHPLC-MS/MS identified 38 compounds. AuNPs-PG showed a plasmon band at 520–550 nm and 88.1 nm hydrodynamic diameter. In SHRs, 0.3 mg/kg AuNPs-PG reduced systolic blood pressure by 18.5% and mean arterial pressure by 15.4%, while mitigating vascular dysfunction by lowering phenylephrine vasoconstriction by 52.7% and boosting acetylcholine vasodilation by 239.3%. Treatment shortened QTc interval by 26.4%, normalized T-wave inversion, and restored renal excretion, increasing urinary volume by 75.0% and sodium by 94.2%. Furthermore, AuNPs-PG elevated placental growth factor by 180.9%, lowered malondialdehyde by 63.5%, and preserved placental histology. Consequently, mean fetal weight increased by 21.6%, implantation index by 50.5%, and offspring-to-mother ratio by 129.6%. Conclusions: AuNPs-PG (0.3 mg/kg) effectively mitigate maternal and fetal complications in gestational hypertension. Gold nanoparticles serve as nanocarriers, whereas surface-adsorbed phytochemicals drive the biological effects.

Pharmaceutics doi: 10.3390/pharmaceutics18091073

Authors: Sophia Leslie Stella Rios Hana Elnahas Megan Keniry

Clinical outcomes for brain cancer are often poor because the blood–brain/tumor barrier hinders effective drug delivery to malignant tissue. Glioblastoma, the most common primary brain malignancy in adults, has an average survival of approximately fourteen months. Here, we discuss novel strategies that our research group and others are developing to deliver chemotherapy to the brain via the nasal cavity. Although significant hurdles remain, intranasal delivery holds substantial promise for improving outcomes for patients with brain cancer. Intranasal delivery is noninvasive, permits repeated dosing, and has been shown to enable direct nose-to-brain transport that bypasses the blood–brain barrier. Challenges such as accurately targeting drugs to the appropriate region of the nasal cavity at therapeutically relevant doses, while maintaining reproducibility, make this cutting-edge approach a regulatory challenge. The prolonged path to clinical translation discourages many researchers from pursuing this potentially life-saving strategy. Nevertheless, preclinical studies demonstrate that intranasal delivery can achieve up to ten-fold higher concentrations of select drugs in the brain. Cancer chemotherapeutics span a wide range of molecular formats, from small molecules to 150-kilodalton antibodies. Accordingly, delivery strategies must be carefully matched to the molecular properties of each therapeutic. Here, we focus on the intranasal delivery of small molecule inhibitors using nanomaterial-based platforms, including aerosols, lipids, gold nanoparticles, gels, emulsions, fibers, and their combinations. Ultimately, we hope that intranasal delivery approaches will be translated to provide patients with better therapeutic outcomes.

Pharmaceutics doi: 10.3390/pharmaceutics18091072

Authors: Nur Aisyah Ikra Nurohman Sriwidodo Sriwidodo Patihul Husni Cecep Suhandi Gofarana Wilar Ahmad Choibar Tridakusumah Sabreena Safuan

Background/Objectives: Mangosteen pericarp extract (Garcinia mangostana L.) exhibits robust antioxidant properties. However, its pharmaceutical application is hindered by poor water solubility and low physicochemical stability. This study aimed to develop a lipid-based nanoemulsion to overcome these limitations and assess its physicochemical characteristics, radical scavenging capacity, and dissolution profiles. Methods: Nanoemulsions were prepared by high-shear homogenization followed by ultrasonication method using sacha inchi oil (Plukenetia volubilis L.) across varying hydrophilic–lipophilic balance (HLB) values. Formulations were characterized by emulsion type, pH, rheology, droplet size, surface charge, encapsulation efficiency, and morphology via transmission electron microscopy. Antioxidant capacity was quantified using the DPPH assay, while in vitro dissolution experiments measured solubility enhancement relative to unformulated extract. Results: All formulations formed stable oil-in-water systems without phase separation. The HLB 10 formulation exhibited optimal performance, yielding a mean droplet diameter of 490.03 ± 32.12 nm, zeta potential of −52.73 ± 0.09 mV, and encapsulation efficiency of 89.14 ± 0.19%. Micrographs revealed well-structured spherical droplets. The raw extract showed strong antioxidant activity (IC50 = 21.68 ppm), which remained functional in nanoemulsion (IC50 = 84.48 ppm). Dissolution testing indicated a 13.4-fold improvement over raw powder. However, storage evaluation indicated chemical loss of bioactive constituents over one month. Conclusions: Sacha inchi oil-based nanoemulsions significantly enhance the dissolution rate of mangosteen pericarp extract while maintaining functional antioxidant potential. Although chemical stability during storage remains a limitation requiring further optimization, this nanocarrier platform offers strong potential to overcome solubility barriers for oral bioactive delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18091070

Authors: Huaying Pan Juan Yang Jiaqi Fu Shenao Yan Yue Yan Yining Xu Minglu Zhou Lian Li Yucheng Xiang

Nanodrug delivery systems have shown great promise against tumors, inflammatory diseases and central nervous system disorders. However, several limitations still exist before further clinical application. The unsatisfactory circulation time in vivo, rapid clearance by the mononuclear phagocyte system, insufficient tissue penetration and low targeting efficiency all hinder the clinical translation of nanomedicines. Cells are the fundamental functional units of the body and their intrinsic biological properties make them promising drug delivery vehicles for targeted therapy. Conjugating cells with nanomedicines combines the carrier functions of living cells with the therapeutic effects of nanodrugs, offering an effective strategy to improve targeted drug delivery. Compared with the existing literature, this review specifically summarizes the biological properties and major applications of six types of living cell carriers. The recent strategies for constructing cell–nanomedicine conjugates are also discussed. Finally, we highlight the current challenges for clinical translation and discuss future directions for the development of cell–nanomedicine delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18091071

Authors: Sandra Miočić Andrea Rašić Jelena Torić Michela Abrami Kristina Ferderber Biserka Cetina-Čižmek Mario Grassi Jelena Filipović-Grčić

Objectives: The physical stability of concentrated azelaic acid nanosuspensions cannot be predicted solely from interfacial stabilisation but reflects the interplay between particle size-dependent thermodynamic driving forces, polymer-mediated rheological structuring and the resulting bulk microstructural organisation. This study employed an A-optimal design of experiments (18 runs) to investigate formulation–process relationships in concentrated azelaic acid nanosuspensions (10–20% w/w) stabilised with a dual HPMC–chitosan system. Methods: Particle size, ζ-potential, rheological behaviour, solid-state properties and in vitro permeation across Strat-M® membranes were evaluated, and formulation–process relationships were analysed using multivariate modelling and logistic regression. Results: Particle size (342–1118 nm; R2 = 0.97) was primarily governed by the applied milling regime. Logistic regression demonstrated a size-dependent probability of crystal growth, with the estimated particle-size transition point (predicted probability = 0.5) shifting from approximately 424 nm after preparation to 594 nm following accelerated storage at 40 °C. CHI concentration controlled ζ-potential (+18.5 to +47.9 mV), although ζ-potential alone did not adequately explain the observed storage stability. Where measurable, zero-shear viscosity (68–45,462 mPa·s) reflected substantial differences in low-shear rheological structuring, while formulations containing higher HPMC concentrations generally exhibited improved stability, consistent with polymer-mediated kinetic constraints on crystal growth. In vitro permeation studies using Strat-M® membranes demonstrated permeation behaviour consistent with structured diffusion-controlled systems, exhibiting lower flux but more uniform permeation profiles than the reference formulation under the applied experimental conditions. Conclusions: Integration of multivariate modelling with rheological, solid-state and permeation characterisation provided an integrated understanding of the formulation–process relationships governing the short-term physical stability and comparative in vitro transport behaviour of concentrated dermal AZA nanosuspensions.

Pharmaceutics doi: 10.3390/pharmaceutics18091069

Authors: Binaya Sapkota Susmita Phuyal Arjun Dhwoj Bamjan Seung-Sik Cho Jung-Hyun Shim Jin Woo Park Laxman Subedi

Topical and transdermal dosage forms can localize therapy or provide controlled systemic exposure, but translation is limited by the selective stratum corneum (SC) barrier and post-application changes in formulation. Existing reviews often address individual enhancers or carriers without integrating drug properties, vehicle microstructure, post-application transformation, quality of evidence, and product development constraints. This review fills that gap by comparing passive formulations, chemical permeation enhancers, vesicular and lipid-based carriers, supersaturating systems, and physical barrier bypass technologies within a skin barrier-informed framework. Increased permeation alone does not establish translational value. Passive delivery remains most feasible for potent, moderately lipophilic small molecules. Chemical enhancers are scalable but limited by irritation and drug-dependent compatibility; nanocarriers may improve solubilization and cutaneous deposition but often lack human confirmation; and physical methods broaden delivery to macromolecules while adding device, manufacturing, usability, and regulatory burdens. Solvent evaporation, residual film composition, supersaturation, precipitation, and drug–vehicle affinity further determine the effective post-application driving force. Accordingly, we propose an evidence-ranked framework linking payload properties and target compartment to mechanism, safety, clinical readiness, and regulatory complexity. It distinguishes mechanistic promise from clinically demonstrated delivery and identifies the evidence needed to advance reproducible topical and transdermal products.

Pharmaceutics doi: 10.3390/pharmaceutics18091068

Authors: Rumeysa Ceylan Eren Aytekin Heybet Kerem Polat Ozan Kaplan Mustafa Çelebier Sibel Bozdağ Pehlivan

Background/Objectives: The aim of this study was to develop and validate a simple, accurate, reproducible, and sensitive reverse-phase high-performance liquid chromatography method for the quantification of reproxalap (RP) in cyclodextrin (CD) inclusion complexes and poloxamer 407 hydrogel formulations. Methods: Chromatographic separation was achieved using a mobile phase of deionized water and an organic phase (methanol:acetonitrile, 55:45 v/v) in a 40:60 (v/v) ratio, at a flow rate of 0.9 mL/min and a run time of 10 min. Validation assessed linearity, specificity, accuracy, and sensitivity over a concentration range of 1–60 μg/mL. Phase-solubility studies were conducted to determine the apparent stability constant (K1:1), and inclusion complexes were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. Results: The method demonstrated specificity, linearity, sensitivity, and accuracy within the tested concentration range of 1–60 μg/mL. The apparent stability constants (K1:1), calculated from the phase-solubility plot slopes, were 2666.66 M−1 for Sulfobutyl Ether β-Cyclodextrin (SBE-β-CD) and 526.3 M−1 for Hydroxypropyl-β-Cyclodextrin (HP-β-CD). The solubility of RP in deionized water (0.25 mM) increased 8.6-fold to 2.15 mM with the HP-β-CD inclusion complex and 22.4-fold to 5.6 mM with the SBE-β-CD inclusion complex. The method was successfully applied to determine RP in inclusion complexes and poloxamer 407 hydrogel formulations, with no interference from formulation excipients. Conclusions: The validated RP-HPLC method is suitable for quality control analysis of RP in CD inclusion complexes and poloxamer 407 hydrogel formulations, supporting its application in the future development of these drug delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18091067

Authors: Lin Zhang Wenbo Yang Shipu Jia Jing Shang Jincheng Wang Xin Zhao Haotian Bai Chenyu Wang

Objectives: Critical-sized bone defects continue to represent a substantial challenge in orthopedic clinical practice. Decellularized bone matrix (DBM) possesses favorable osteoconductive properties due to its retention of native extracellular matrix architecture and collagen components. However, its limited osteogenic bioactivity restricts its application in complex bone defect repair. This study aimed to construct a dendritic mesoporous silica (DMSN)-modified DBM composite scaffold loaded with teriparatide (DBM-DMSN@TPTD) and to systematically evaluate its physicochemical properties, drug release behavior, biocompatibility, and osteogenic differentiation-promoting capacity. Methods: A DBM scaffold was prepared from bovine femoral cancellous bone via a combined freeze–thaw and chemical detergent decellularization method. DMSNs were synthesized through a sol–gel method, amine-functionalized with APTES, and covalently grafted onto the DBM surface via EDC/NHS crosslinking. Teriparatide was loaded onto the composite scaffolds at three concentrations (1, 10, and 100 nmol/L). The scaffolds were characterized via SEM, TEM, BET, EDS and XPS. Decellularization efficacy was assessed by DAPI staining and nucleic acid quantification. Drug release behavior was evaluated through in vitro release studies, while biocompatibility and osteogenic differentiation of rat BMSCs were examined using Live/Dead staining, phalloidin/DAPI cytoskeletal staining, CCK-8 assays, ALP staining, and RUNX2/OCN immunofluorescence. Results: DMSNs demonstrated a dendritic mesoporous architecture, featuring a specific surface area of 390.44 ± 1.78 m2/g and pore diameters within the range of 15–20 nm. DBM showed effective removal of immunogenicity, with well-preserved collagen architecture. Drug release displayed a biphasic pattern, with 56.03% released within the first 72 h and 83.23% by day 16. None of the tested scaffolds showed obvious cytotoxicity under the experimental conditions. The DBM-DMSN@TPTD-M group (10 nmol/L) produced the strongest effects on BMSC proliferation and osteogenic differentiation, as indicated by the highest ALP activity and elevated RUNX2 and OCN expression (p < 0.05). Conclusions: The DBM-DMSN@TPTD scaffold offers a native bone microenvironment, sustained drug release, and osteogenic activity in vitro. These features may support BMSC proliferation and osteogenic differentiation. Accordingly, this scaffold warrants further investigation as a potential strategy for bone defect repair.

Pharmaceutics doi: 10.3390/pharmaceutics18091066

Authors: In Gyu Yang Jun-Young Han Min Young Jeong Dong-Wan Seo Myung Joo Kang Sun Ho Kim

Objectives: Fixed-dose combination tablets containing sitagliptin hydrochloride (SG) and metformin hydrochloride (MF) are a mainstay in the clinical management of type 2 diabetes. However, SG is highly susceptible to chemical degradation during storage, particularly in the presence of MF. Herein, a bilayer tablet in which SG and MF are physically separated into distinct layers was designed to enhance the chemical stability of SG while ensuring pharmacokinetic equivalence to the marketed reference product Janumet®. Methods: The compositions of individual SG and MF compartments were selected based on evaluations of their physical properties and dissolution profiles. Critical process parameters, including the pre- and main compression forces and coating levels of bilayer tablets, were fine-tuned to achieve dissolution characteristics comparable to those of the reference product. Results: Under accelerated storage conditions (40 °C, 75% relative humidity), the optimized bilayer tablet exhibited superior stability, with total SG-related impurity levels of 0.18% compared with 0.86% in the reference product after six months. Furthermore, in a randomized bioequivalence study in healthy volunteers (n = 30), the SG/MF bilayer tablet was pharmacokinetically equivalent to the reference product, with all parameters falling within the Food and Drug Administration-mandated regulatory criteria. Conclusions: In conclusion, this SG/MF bilayer tablet has better storage stability than the reference product and may be an alternative to conventional SG/MF combination tablets.

Pharmaceutics doi: 10.3390/pharmaceutics18091065

Authors: Moataz M. Rashad Shaimaa M. Kasem Khaled E. El-Kelany Sara A. Abdel Gaber

Purpose: Cryptosporidiosis is a parasitic inflammatory disease that causes the death of around 1.6 million people annually worldwide and is associated with Cryptosporidium parvum (C. parvum) infection. This study aimed to evaluate the therapeutic efficacy of carvacrol (CV) loaded into mesoporous silicate nanoparticles (MSNs) against C. parvum using in vitro, in vivo, histopathological, immunohistochemical, biochemical, and computational approaches. Methods: In vitro assays were conducted to evaluate the oocysticidal activity of CV-loaded MSNs (CV-MSNs) for 96 h. In silico assays were carried out to investigate target inflammatory proteins involved in cryptosporidiosis. In vivo tests were performed on male Swiss albino mice (22 ± 5 g, 4–6 weeks) to assess the anti-inflammatory and antioxidant effects of CV after loading onto MSNs. Results: CV showed its highest oocysticidal efficacy after loading onto spherical 72 nm MSNs (MSN2), exhibiting a 0.07 mg/mL LC50. At the in vivo level, CV-MSN2 markedly restored ileal, pulmonary, and hepatic histoarchitecture and normalized biochemical indices in infected mice. CV-MSN2 also showed the strongest inhibitory action against computationally selected target proteins, TLR-4, NF-κb-P65, STAT-3, NOS2, and JAK-1, with accompanying downregulation of IL-6, IL-1β, and TNF-α. Similarly, antioxidant markers were significantly decreased following CV-MSN2 treatment compared to the positive controls. Conclusions: This study shows that MSNs enhanced CV efficacy against ileal inflammation induced by C. parvum, preserving CV’s molecular targets.

Pharmaceutics doi: 10.3390/pharmaceutics18091064

Authors: Noelia Fernández-Ceballos Laura Gallego-Yerga Rafael Peláez

Background: Antimitotic agents are very successful antitumor therapies, but lack tumor selectivity, causing toxicity. Antitumor alkylphospholipids (APLs) selectively accumulate in tumor cells but display low potencies. Hypothesis: Incorporating APL moieties onto antimitotic N-trimethoxyphenyl naphthalene sulfonamides (TMNS) might afford SMDCs with the potency of antimitotics and the tumor selectivity of APLs. Methods: 24 new TMNSs with spacers of different lengths (4 to 9 atoms) and nature (alkanes or ethers) on the sulfonamide nitrogen and capped with phosphorus-containing groups such as diethylphosphonates, phosphonic acids, and hydrogenophosphonate esters of aminoalcohols (diethylaminopropanol, choline, or dimethylaminoethanol) were designed and synthesized. Their antiproliferative effects against several cancer cell lines and their cotreatment with verapamil to assess whether they are substrates of MDR pumps were evaluated. The mechanism of action was studied: cell cycle effects, apoptosis induction, and immunofluorescence microscopy. Computational studies considered binding to tubulin and pharmacokinetics. Results: Diethyl phosphonates and phosphonic acids are antiproliferative in the micromolar to submicromolar range. P-gp inhibitor verapamil renders inactive compounds active, suggesting that efflux, not binding, removes activity. Mechanistic studies agree with an antimitotic action. Proposed binding to tubulin is similar to TMNS, with the phospholipid-like substituent projecting towards the interdimer space. Conclusions: Hybridization of TMNSs with potentially tumor–targeting APLs yields microtubule-disrupting antitumor compounds. However, the modifications assayed turn the compounds into substrates of MDR. These compounds are a proof of concept of the strategy that might succeed if future modifications avoid MDR and might target the compounds towards cancer cells.

Pharmaceutics doi: 10.3390/pharmaceutics18091063

Authors: Saif-alnasr H. Mohammed Ayman B. Mousa Mohammed Taj-Eldin Abdalla Anita Sztojkov-Ivanov Kálmán F. Szűcs Róbert Gáspár

Background: Sex hormones play crucial functions in the body via the genomic and non-genomic pathways. 5α- and 5β-dihydrotestosterone (5α- and 5β-DHT) are 5-reduced testosterone metabolites. We aimed to investigate the non-genomic effect of 5α- and 5β-DHT on uterine muscle contractility in vitro and in vivo for non-pregnant and 22-day-pregnant rats. Methods: The rapid in vitro action of 5α-DHT and 5β-DHT (10−9–10−3 M) on KCl (25 mM)-stimulated contractions was examined in an organ bath in the presence of several blockers and after endometrium removal. The actions of DHTs (10−4 M) and nifedipine (10−7 M) were also examined in contractions stimulated by KCl (40 mM) with a cumulative addition of CaCl2 (3–120 mM). Plasma DHT levels were measured by ELISA after a single intraperitoneal (i.p.) administration of DHT (10 mg/kg), and kinetic curves were obtained. The in vivo relaxing action of DHTs was detected by strain-gauge sensors. The animals received 5α- or 5β-DHT alone (3/10/30/100/300 mg/kg i.p.) or with flutamide (100 mg/kg i.p.). Results: DHT showed concentration-dependent relaxation of uterine muscle in vitro, with a higher potency observed for 5β-DHT. Among the blockers used, G15 and L-NAME reduced the potency of 5α-DHT in pregnant rats only. Both DHTs inhibited the contraction-increasing effect of CaCl2, proving their Ca2+ inhibiting effects. DHTs had similar cmax and tmax values in both non-pregnant and pregnant rats. DHT plasma levels before and 30 min after administration were proportional to the administered doses. Their single doses (30/100/300 mg/kg) elicited a flutamide-resistant uterine relaxing effect in vivo. Conclusions: DHTs or their analogs are candidates for further studies in the human uterus to establish their potential to treat conditions associated with uterine hyperactivity.

Pharmaceutics doi: 10.3390/pharmaceutics18091062

Authors: Eunseok Jang Gaeun Lee Yoseph Seo Hyunjun Park Suk Min Yun Sang Deuk Lee Giwon Lee Chulhwan Park Taek Lee

The biological fate of lipid-based nanocarriers (LBNs) is shaped at the interface. Whereas core architecture governs cargo loading, protection, and baseline release, surface architecture mediates the carrier’s initial interactions with proteins, cells, extracellular matrices, and tissue barriers, thereby influencing colloidal stability, immune recognition, targeting, biodistribution, barrier transport, and release initiation. Layer-by-layer (LbL) engineering provides a modular strategy for programming this interface through sequentially assembled coatings in which functional components are spatially separated yet mechanistically coordinated. By integrating polymers, biomolecules—including peptides and nucleic acids—and stimuli-responsive materials, LbL systems can decouple functions that are difficult to regulate independently within conventional single-layer or compositionally mixed surface architectures. This review examines recent advances in LbL-engineered LBNs (LbL-LBNs), focusing on how multilayer surface architecture reshapes physicochemical properties, cargo localization and release, biological identity, cellular interactions, and transport across physiological barriers. Particular attention is given to the multilayer interface as a dynamic biointerfacial bridge between a cargo-specific core architecture and the surrounding biological environment, including its capacity for stimuli-responsive switching in pathological microenvironments. The discussion further extends to biomimetic hybrid interfaces and establishes a framework for translating hierarchical surface architectures into reproducible, clinically tractable platforms for precision therapeutic delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18091061

Authors: Katarzyna Reczyńska-Kolman Jan Grabiński Konrad Kwiecień Dorota Ochońska Kinga Pielichowska Monika Brzychczy-Włoch Elżbieta Pamuła

Background/Objectives: Targeted pulmonary delivery of antibiotics offers a favorable approach for the treatment of lower respiratory tract infections (LRTIs) by enhancing local drug concentrations while minimizing systemic exposure. This study aimed to develop and characterize fatty acid-based nanostructured lipid carrier (NLC) microparticles, including tobramycin-loaded formulations, and assess their potential application as carriers for pulmonary drug delivery. Methods: Tobramycin-loaded NLC microparticles were prepared using a hot emulsification method, with lauric acid as the solid lipid and oleic acid as the liquid lipid. The obtained formulations were characterized in terms of morphology, particle size distribution, zeta potential, thermal properties, encapsulation efficiency, drug-release behavior, aerodynamic properties, and powder flowability. Antibacterial activity was evaluated against S. aureus and P. aeruginosa, while cytocompatibility was assessed using BEAS-2B human-lung epithelial cells. Results: The developed microparticles exhibited spherical morphology, with particle characteristics influenced by lipid composition. Incorporation of oleic acid increased tobramycin encapsulation efficiency, reaching approximately 75% at liquid lipid concentrations of 5–15%. Theoretical aerodynamic diameters remained within the respirable range, and increasing oleic acid content improved powder flowability. Drug-release behavior was strongly dependent on lipid composition, with the optimized formulation providing prolonged tobramycin release over 72 h. All formulations preserved antibacterial activity against S. aureus and P. aeruginosa and demonstrated good cytocompatibility with BEAS-2B cells. Conclusions: Fatty acid-based nanostructured lipid microparticles represent a promising active carrier platform for the pulmonary delivery of tobramycin. The developed system demonstrated efficient drug encapsulation, satisfactory particle characteristics, controlled drug release, preserved antibacterial activity, and good biocompatibility, making them a suitable component of dry powders for inhalation (DPI).

Pharmaceutics doi: 10.3390/pharmaceutics18091060

Authors: Archana Krovi Leanna Levin Greg J. Gatto Ellen H. Luecke Rhonda Brand Amanda Swistok Mackenzie L. Cottrell Amanda P. Schauer Leah M. Johnson

Background/Objectives: Multipurpose prevention technologies (MPTs) that combine protection against HIV and unintended pregnancy can improve women’s health outcomes by simplifying use and enhancing adherence. We developed a long-acting (LA) implant for the simultaneous and sustained delivery of the antiretroviral islatravir (ISL) and the contraceptive hormone etonogestrel (ENG). Methods: Using extruded poly-ε-caprolactone (PCL) tubing, reservoir-style implants were fabricated and evaluated in three configurations: (Group 1) a two-segment implant formed by joining separate ISL- and ENG-containing tubes; (Group 2) a single tube divided into two drug compartments by a heat-sealed PCL spacer; (Group 3) separate single-drug implants. In vitro release was assessed in phosphate-buffered saline at 37 °C, and safety and pharmacokinetic (PK) profiles were evaluated in New Zealand White rabbits (n = 4/group) over approximately 90 days. Results: The average ISL daily in vitro release rates for Groups 1, 2, and 3 were 29 ± 5 µg/day, 18 ± 5 µg/day, and 47 ± 8 µg/day, respectively, and ENG daily in vitro release rates were 39 ± 9 µg/day, 41 ± 8 µg/day, and 33 ± 7 µg/day. From day 28 through the end of the study, median (IQR) plasma concentrations were 0.22 (0.18–0.27) ng/mL, 0.28 (0.24–0.33) ng/mL, and 0.31 (0.28–0.33) ng/mL for ISL in Groups 1, 2, and 3, respectively, and 0.26 (0.21–0.31) ng/mL, 0.25 (0.16–0.47) ng/mL, and 0.30 (0.23–0.38) ng/mL for ENG in Groups 1, 2 and 3, respectively. Implants across all groups were well tolerated and demonstrated favorable local tolerability over the 90-day dosing period. Conclusions: Integration of multiple indications into a single platform capable of sustained release over an extended duration holds potential to address persistent gaps in women’s sexual and reproductive health needs.

Pharmaceutics doi: 10.3390/pharmaceutics18091059

Authors: Philippe Espeau

The context of 3D printing in the pharmaceutical sector and its significance [...]

Pharmaceutics doi: 10.3390/pharmaceutics18091058

Authors: Barbara Zawidlak-Węgrzyńska Joanna Rydz

Polymeric drug delivery systems continue to play a pivotal role in modern pharmaceutical research, offering innovative solutions for overcoming limitations associated with conventional drug administration [...]

Pharmaceutics doi: 10.3390/pharmaceutics18091057

Authors: Ravi Maharjan Ivana D’Angelo

The respiratory tract is one of the most fascinating yet challenging routes for drug delivery [...]

Pharmaceutics doi: 10.3390/pharmaceutics18091056

Authors: Luca Éva Uhljar Zsófia Viktória Tagscherer Rita Ambrus

Background/Objectives: Electrospun polyvinylpyrrolidone (PVP) fibers are highly promising for fast-dissolving drug delivery. Methods: In this study, five PVP grades with molecular weights ranging from 4000 to 1,300,000 Da were investigated over a broad concentration range (10–60 w/w%) to evaluate their electrospinnability and dissolution behavior. Results: A well-defined viscosity window of approximately 150–680 mPa·s was identified for the formation of continuous, bead-free fibers. Deviations from this optimal window resulted in electrospraying or jet instability. Continuous fibers were successfully prepared from all investigated PVP grades, including low-molecular-weight PVP K-12 (Mw 4000), demonstrating that appropriate solution viscoelasticity can compensate for limited chain entanglement. Remarkably, in vitro testing revealed that all fibrous formulations exhibited ultrafast disintegration (0.29–1.33 s) and dissolution (0.39–2.32 s). Statistical analysis confirmed no significant differences attributable to polymer molecular weight or fiber diameter, effectively challenging the common assumption that higher-molecular-weight PVP delays disintegration. Instead, the immediate dissolution originates from rapid wetting and capillary-driven fluid uptake, facilitated by the highly porous nano- and microfibrous network. Conclusions: By highlighting the dominant role of macroscopic structural properties over polymer chain length, these findings provide a practical framework for the development of fast-dissolving electrospun PVP-based drug delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18091055

Authors: Anastassiya Khrustaleva Azamat Yedrissov Dmitriy Khrustalev Ivan Chernykh Aleksandr Samorodov Saule Akhmetova Artyom Savelyev Marlen Kiikbayev Polina Rusyaeva Vladimir Kazantsev Kristina Perepelitsyna Sofiya Shapovalenko

Background/Objectives: The development of multifunctional biomaterials for bone regeneration remains a key challenge in additive manufacturing. Although polylactic acid (PLA) is widely used in fused deposition modeling (FDM), its limited bioactivity and lack of intrinsic antibacterial functionality restrict its application in implantable constructs. This study aimed to develop a PLA-based composite filament combining ion-mediated bioactive potential and local antibacterial functionality using a Quality by Design (QbD) approach. Methods: PLA-based composite filaments incorporating a mollusk shell-derived biogenic calcium-containing filler (20 wt.%) and gentamicin (5 wt.%) were fabricated by solvent-free melt extrusion. A QbD framework was applied to define the Quality Target Product Profile (QTPP), identify critical quality attributes (CQAs), and assess critical material attributes (CMAs) and critical process parameters (CPPs). The material was characterized by SEM–EDS combined with ImageJ-based quantitative image analysis, TGA/DSC, mechanical testing, ICP-AES analysis of aqueous extracts, agar diffusion antibacterial assays, FDM printability assessment, and in vivo biocompatibility testing in a rat subcutaneous implantation model. Results: The developed PLA–Gen–MS material was obtained as a continuous filament with a diameter of 1.75 ± 0.05 mm and was successfully used for FDM printing of model scaffold structures. SEM–EDS confirmed matrix continuity and distribution of the calcium-containing mineral phase. ICP-AES revealed a calcium-dominant multicomponent ion release profile, with Ca as the predominant element and measurable levels of Sr, Mg, P, Mn, and Fe. TGA/DSC confirmed thermal compatibility of the components under melt-processing conditions. PLA–Gen–MS demonstrated antibacterial activity against all tested strains, with inhibition zones of approximately 20–21 mm. In vivo, the material showed a favorable preliminary tissue response compared with TiLOOP®, including faster reduction of inflammatory infiltration and absence of foreign body giant cells by day 14. Conclusions: The QbD-guided strategy enabled the development of a multifunctional PLA-based filament integrating melt processability, structural integrity, ion-mediated bioactive potential, antibacterial functionality, printability, and favorable preliminary biocompatibility. PLA–Gen–MS can be considered a promising platform for further development of personalized bioactive and antibacterial scaffold constructs for bone regeneration.

Pharmaceutics doi: 10.3390/pharmaceutics18091054

Authors: Akshay Kumar Devesh Kumar Mohit Agrawal Jaspreet Kaur Mohit Kumar Dinesh Kumar Neeraj Choudhary Thakur Gurjeet Singh Ankit Awasthi Emad M. Abdallah

Wound healing is a dynamic and highly coordinated process that involves inflammation, cell proliferation, angiogenesis, re-epithelialization, extracellular matrix remodeling, and tissue maturation. The altered expression of important signaling pathways, such as transforming growth factor-β (TGF-β)/Smad, nuclear factor-κB (NF-κB), phosphoinositide 3-kinase/protein kinase B (PI3K/Akt), mitogen-activated protein kinase (MAPK), and Wnt/β-catenin, may be responsible for slower wound healing, chronic inflammation, excessive fibrosis, and impaired tissue regeneration. Multifunctional nanomaterials are a promising strategy for tuning these highly coordinated processes due to their tunable physicochemical properties, high surface area, and the ability to deliver cargo, as well as the integration of antimicrobial, antioxidant, anti-inflammatory, and pro-angiogenic properties. The aim of current review is to summarize the potential of multifunctional nanomaterials to promote wound healing, with a focus on mechanisms of action and modulation of key cellular signaling pathways. A systematic review of the literature was conducted using PubMed, Scopus, Web of Science, and Google Scholar, searching for publications from 1996 to June 2026, and representative experimental, mechanistic, preclinical, and translational studies were critically evaluated. In this review, the authors discuss the role of nanomaterial properties, therapeutic payload, molecular targets, modulation of cellular signaling pathways, and regenerative effects. These platforms have been shown in in vitro and animal studies to influence inflammatory signaling, oxidative stress, angiogenesis, collagen remodeling, re-epithelialization, cellular proliferation, and migration. However, the modulation of these pathways are dose-responsive, time-dependent, and cell- and wound-stage-specific. Despite the promising therapeutic potential of nanomaterial-based wound care strategies, the available evidence remains predominantly preclinical, with relatively limited clinical data supporting their use in humans. Concerns regarding long-term toxicity, biodistribution, batch-to-batch reproducibility, sterilization, scalable manufacturing, regulatory approval, and commercial feasibility further challenge translation into clinical practice. Multifunctional nanomaterials may offer a promising approach for pathway-specific and multimodal wound management; however, comprehensive mechanistic studies, long-term safety and biodistribution assessments, and well-designed clinically relevant investigations are required to establish their efficacy, safety, and true translational potential.

Pharmaceutics doi: 10.3390/pharmaceutics18091053

Authors: Arianna Minzoni Benjamin Bobay Shriarjun Shastry Eduardo Barbieri Brandon Brino Crystal Collazo Shizuo Kamita Danni Wang Ciera Khuu Alexander Polgar Joseph Siino Sushmita Koley Peyton Russelburg Mark Snyder Christopher Belisle Michael Daniele Stefano Menegatti

Background/Objectives: Adeno-associated virus (AAV) capsids undergo pH-dependent conformational gating at the 5-fold symmetry pore, but how these structural dynamics shape serotype-specific behavior and affinity-ligand recognition remains unclear, particularly for the clinically important serotypes AAV8 and AAV9. This study aimed to establish a pH-resolved structural framework linking 5-fold pore dynamics to peptide-ligand recognition and to translate this framework into sequence-based design principles for affinity capture of gene therapy vectors. Methods: AAV8 and AAV9 5-fold capsid assemblies were subjected to 500 ns molecular dynamics simulations under acidic (pH 5), neutral (pH 7), and basic (pH 9) conditions, with analysis of pore volume, inter-residue contact networks, electrostatic potential, and solvent-accessible surface area. In parallel, affinity chromatography using three mixed-mode peptide ligands (RVVAVYRI, TTFRAHHI, and TYHHHHII) was performed on clarified HEK293 lysates containing AAV8 or AAV9, with capsid yield, host-cell-protein clearance, and transduction activity assessed by ELISA, SEC-HPLC, and flow-cytometry-based transduction assays. Results: AAV8 displayed a heterogeneous, bimodal pore conformational landscape at pH 7, whereas AAV9 exhibited a discrete gate-like transition with maximal pore constriction at physiological pH; both serotypes showed pore-proximal contact remodeling with distinct network topologies. Experimentally, TYHHHHII achieved the highest selectivity for genome-containing capsids at pH 7, with transduction activity enrichment factors of 2.82 (AAV8) and 5.61 (AAV9), while TTFRAHHI provided the broadest operational pH range for bulk capsid recovery. Conclusions: These findings establish a structural framework linking pH-dependent pore dynamics to affinity ligand recognition and suggest practical sequence-design rules for ligand engineering: clustered histidines for neutral-pH selectivity, Arg-containing motifs for broad-pH robustness, and aromatic or hydrophobic residues for reinforcement of capsid binding.

Pharmaceutics doi: 10.3390/pharmaceutics18091052

Authors: Kulzhan Berikkhanova Alexandr Gulyayev Yernur Zakirov Askhat Zhilkaidarov Azhar Zhaisanova Nurgul Daniyeva Ardak Omarbekov Gulsara Berikkhanova Yessenkhan Sultan Zhannat Zhakiyanova Gulyash Tanysheva

Background/Objectives: Ceftriaxone (Ctx) is a third-generation cephalosporin widely used to treat infections caused by Gram-positive and Gram-negative bacteria. However, its clinical efficacy may be limited by rapid systemic elimination and suboptimal tissue distribution. Erythrocyte-based targeted drug delivery systems (TDDSs) have emerged as a promising approach to prolong drug circulation and enhance site-specific accumulation. This study investigated the pharmacokinetic profile and tissue distribution of ceftriaxone encapsulated in autologous erythrocytes (RBC-Ctx) compared with free ceftriaxone (Free-Ctx) following intravenous administration in rats. Methods: Ceftriaxone was encapsulated into autologous rat erythrocytes using a hypoosmotic hemolysis loading technique. Drug-loaded erythrocytes are called pharmacocytes. Adult male Wistar rats received a single intravenous injection of Free-Ctx or RBC-Ctx at an equivalent ceftriaxone dose of 340 mg/kg. Plasma samples were collected over 24 h for pharmacokinetic analysis, while the liver, spleen, lungs, kidneys, heart, pancreas, and skeletal muscle were harvested at 1 and 12 h for tissue distribution studies. Ceftriaxone concentrations were quantified by high-performance liquid chromatography with UV detection. Results: Erythrocyte encapsulation significantly modified the pharmacokinetic behavior of ceftriaxone. Compared with Free-Ctx, RBC-Ctx prolonged the elimination half-life (4.4 ± 0.6 vs. 1.8 ± 0.1 h), increased systemic exposure (AUC0–last, 1.6 ± 0.1 vs. 1.2 ± 0.2 mg·h/mL), reduced total body clearance (218.2 ± 11.0 vs. 294.0 ± 48.8 mL/h/kg), and increased the apparent volume of distribution at steady state (688.3 ± 61.0 vs. 435.0 ± 23.1 mL/kg). In addition, RBC-Ctx was associated with a distinct relative tissue-distribution pattern of ceftriaxone, particularly in reticuloendothelial system-rich organs such as the liver and spleen, while ceftriaxone remained detectable in several tissues at 12 h after administration. In contrast, ceftriaxone concentrations following Free-Ctx declined markedly or became undetectable over the same period. Conclusions: Encapsulation of ceftriaxone into autologous erythrocytes substantially prolonged systemic circulation, enhanced drug exposure, reduced clearance, and altered the relative tissue distribution of ceftriaxone. These findings demonstrate that erythrocyte-based carriers effectively modulate ceftriaxone pharmacokinetics and tissue distribution, supporting their potential as a targeted antibiotic delivery platform for improving antimicrobial therapy, particularly for infections involving reticuloendothelial system-associated tissues. Further studies in experimental models of infection and inflammation are warranted to evaluate therapeutic efficacy under pathological conditions and to optimize this delivery strategy.

Pharmaceutics doi: 10.3390/pharmaceutics18091051

Authors: Hui Liu Ting Li Xinlei Chen Hongling Yu Yuchun Men Huiwen Tan Jiaqi Li Yerong Yu

Background/Objectives: Standard protocols for euglycemic clamp studies involving long-acting insulin formulations typically recommend a dosage range of 0.4~0.6 U/kg for a single dose. This investigation aimed to evaluate the bioequivalence of an insulin degludec (IDeg) biosimilar versus its reference product, while simultaneously analyzing intra-subject variability in pharmacokinetic (PK) and pharmacodynamic (PD) metrics at doses of 0.4 U/kg and 0.5 U/kg in healthy Chinese adults. Methods: This randomized, single-dose, crossover euglycemic clamp study involved 52 participants who received either 0.4 U/kg (Group A, n = 26) or 0.5 U/kg (Group B, n = 26) of both formulations. Key outcomes measured included AUCIDeg,0–24h, Cmax,IDeg, and AUCGIR,0–24h. Secondary endpoints encompassed AUC of IDeg or GIR at specified intervals and time-related metrics. Intra-subject variability (intra-CV) was assessed for PK/PD parameters. Results: All subjects completed the study without dropping out. Baseline demographics showed no significant disparities between groups. Bioequivalence criteria were satisfied for all PK/PD endpoints, with 90% confidence intervals (CIs) falling within the 0.80~1.25 range, with the exception of AUCGIR,0–12h at the 0.4 U/kg dose (90% CI: 0.944~1.333). Notably, the 0.4 U/kg dose exhibited significantly higher intra-CV for AUCGIR,0–24h (24.9% vs. 20.2%, p = 0.049) and AUCGIR,0–12h (37.6% vs. 27.6%, p = 0.005) compared to the 0.5 U/kg dose. Conclusions: Applying a 0.5 U/kg dose reduced intra-subject PD variability and improved equivalence likelihood for secondary endpoints, indicating that a higher single dose might be optimal in bioequivalence study design.

Pharmaceutics doi: 10.3390/pharmaceutics18091050

Authors: Hanan Aati Jawaher H. Alqahtani Areej Al-Taweel Sultan Y. Aati

Background/Objectives: Propolis is a resinous natural product rich in phenolic acids and flavonoids, recognized in ethnopharmacology for its antioxidant and anti-inflammatory properties. This study aimed to identify the most bioactive Saudi propolis extract using a bioassay-guided strategy and investigate its chemical profile and mechanistic anti-inflammatory potential. Methods: Four propolis extracts (P1–P4) collected from different regions of Saudi Arabia were evaluated for their antioxidant (DPPH and ABTS) and anti-inflammatory (COX-1 and COX-2) activities. The most active extract was profiled using liquid chromatography–mass spectrometry (LC–MS), followed by molecular docking and molecular dynamics simulations. Results: Among all samples, P3 demonstrated the strongest antioxidant activity, with IC50 values of 25.84 ± 0.96 µg/mL (DPPH) and 32.30 ± 1.20 µg/mL (ABTS), comparable to ascorbic acid (27.45 ± 1.42 and 21.22 ± 0.79, respectively). P3 also exhibited potent and selective COX-2 inhibition with an IC50 of 6.19 ± 0.21 µg/mL (relative to celecoxib, IC50 0.681 ± 0.02 as positive control). LC–MS analysis identified 26 secondary metabolites. Computational studies ranked kaempferol as forming the most conformationally stable COX-2 complex among the ligands examined, including the reference inhibitor, on the basis of molecular dynamics descriptors of pose persistence and conformational confinement. Conclusions: Saudi propolis P3 is a potent source of bioactive compounds with strong antioxidant and selective COX-2 inhibitory activity. These findings highlight its anti-inflammatory potential and suggest its value as a natural lead for developing safer therapeutics targeting inflammation-related chronic diseases.

Pharmaceutics doi: 10.3390/pharmaceutics18091049

Authors: Alba Escolà-Rodríguez Elena Sandoval Jorge Moisés Adrián Téllez Santoyo Albert Carramiñana Jaime I. Sainz de Medrano Cristina Espinosa Carlos Roca Marta Hernández Meneses Sabina Herrera Mercè Brunet Serra Pedro Castro Dolors Soy Muner Carla Bastida

Background: Isavuconazole, a broad-spectrum triazole antifungal, exhibits high lipophilicity and extensive plasma protein binding, properties that may predispose it to sequestration within extracorporeal membrane oxygenation (ECMO) circuits. This study aimed to characterize the pharmacokinetics (PK) of isavuconazole and to evaluate drug sequestration within the ECMO circuit in critically ill patients receiving ECMO support. Methods: We conducted a prospective, exploratory, single-center observational study including critically ill patients receiving ECMO (veno-venous (VV) or veno-arterial (VA)) and treated with intravenous isavuconazole. Serial blood samples were collected simultaneously from the patient’s arterial line and from pre- and post-membrane oxygenator sampling sites. Non-compartmental analysis was performed on arterial line samples to estimate PK measures, and concentration differences across sampling sites were analyzed to estimate circuit-related drug loss. PK/pharmacodynamic (PD) target attainment was assessed using established efficacy thresholds (AUC0–24/MIC ≥ 25 and Cmin > 2 mg/L). Results: A total of 41 plasma samples from 3 critically ill patients (2 VV-ECMO, 1 VA-ECMO) were included in the analysis. Limited, component-specific isavuconazole loss was observed in tubing and connectors (6.62% ± 20.8%, p = 0.294) and across the entire ECMO circuit (7.74% ± 20.2%, p = 0.211). Likewise, no relevant concentration difference was detected across the membrane oxygenator (0.849% ± 6.15%, p = 0.642). Interindividual variability was observed across PK parameters, particularly in measures of elimination and distribution. All patients achieved predefined PK/PD efficacy targets, with mean Cmin and AUC0–24/MIC of 3.07 ± 0.261 mg/L and 85.3 ± 4.03, respectively, and none exceeded the established toxicity threshold. Conclusions: Preliminary results showed variable concentration differences across ECMO sampling sites, with no consistent pattern of isavuconazole loss across the ECMO circuit under the conditions evaluated. All patients achieved predefined PK/PD efficacy targets using currently recommended dosing regimens; however, interindividual PK variability was observed, supporting the potential value of therapeutic drug monitoring (TDM) to guide individualized dosing decisions in this population. Larger population PK studies are warranted to further characterize determinants of isavuconazole exposure during ECMO support and refine evidence-based dosing strategies.

Pharmaceutics doi: 10.3390/pharmaceutics18091048

Authors: Adeola Aminu Zhenjia Wang

Acute and chronic inflammation underlies the pathogenesis of numerous diseases, including autoimmune disorders, atherosclerosis, infections, and cancer. Although non-steroidal anti-inflammatory drugs (NSAIDs) and corticosteroids are widely used to control inflammation, their clinical utility is limited by adverse effects such as gastrointestinal toxicity and immunosuppression. Specialized pro-resolving mediators (SPMs), a family of endogenous lipid mediators derived from omega-3 fatty acids, have emerged as promising therapeutics because they actively promote the resolution of inflammation without suppressing host immunity. However, their clinical translation is hindered by poor chemical stability, rapid metabolic degradation, and short circulation half-lives. To overcome these limitations, a variety of delivery platforms—including liposomes, extracellular vesicles, PLGA nanoparticles, and hydrogels—have been developed to improve SPM stability, pharmacokinetics, and therapeutic efficacy. This review summarizes the cellular targets of SPMs, current delivery challenges, and emerging strategies for cell- and tissue-specific SPM delivery. We also discuss future opportunities for targeted SPM therapies in the treatment of inflammatory diseases and cancer.

Pharmaceutics doi: 10.3390/pharmaceutics18091047

Authors: Monika Michalak

Background: Plants have been traditionally used for centuries to treat wounds and, over time, have been tested for their healing properties. There is a constant search for new natural resources that could be used to develop various topical wound care products. Methods: A comprehensive search of the literature was conducted in PubMed/MEDLINE, Scopus, and Web of Science databases (2022–2026) in accordance with the PRISMA 2020 guidelines. Included studies focused on current in vitro and in vivo research on hydrogels containing plant extracts from the Asteraceae and Lamiaceae families and their potential application in wound healing and skin regeneration. Results: An analysis of 24 included studies confirms that both families include interesting and valuable plants with antioxidant, antimicrobial and anti-inflammatory properties; these plants also influence collagen deposition, fibroblast proliferation, and epithelialization, and reduce the risk of infection, thereby contributing to faster wound healing. The most frequently tested phytoextract in this respect was Calendula officinalis (Asteraceae) incorporated into a hydrogel. A variety of materials, including natural, semi-synthetic, and synthetic polymers, as well as hybrid matrix, but also diverse formulation strategies, from simple solutions to more advanced and modern methods, have been used to produce hydrogels. Conclusions: This systematic review summarizes the current evidence, highlights directions and possibilities for the use of phytoextract-based hydrogels, and discusses limitations and future perspectives in the development of effective externally applied formulations to support wound healing.

Pharmaceutics doi: 10.3390/pharmaceutics18091046

Authors: Christian Fleck Isameddin Aghrbi Kristina Vlahovic Franciska Erdő András József Laki Nikolett Kállai-Szabó István Antal Miléna Lengyel

Background/Objectives: Matrix pellet formulations enable homogeneous incorporation of the active ingredient and advanced control of release depending on the excipients added. The aim of this study was to develop, characterize, and assess the drug release profiles of microcrystalline cellulose (MCC) and isomalt-based matrix pellets with direct drug incorporation, advancing beyond the earlier concept of isomalt as a mere inert core with layered drug application, under simulated physiological conditions in vitro. Methods: Matrix pellets with varying MCC–isomalt ratios (90:10, 70:30, and 50:50) were produced by extrusion/spheronization and subsequently coated with film-forming polymers. Dissolution experiments were performed under varying osmolarity to characterize release profiles. The experimental design data were statistically evaluated. Results: The extrusion/spheronization technique yielded uniform, robust matrix pellets with acceptable sphericity and mechanical integrity, even at high isomalt levels. Release from coated pellets was significantly influenced by the polymer coating and osmolarity of the dissolution medium. However, with increasing isomalt content in the matrix, the dependence of drug release kinetics on medium osmolarity was substantially reduced. Conclusions: The production of MCC–isomalt matrix pellets in which isomalt acts as a functional matrix component reduced the effect of osmolarity of dissolution medium on the release of ibuprofen sodium salt in in vitro experiments.

Pharmaceutics doi: 10.3390/pharmaceutics18091045

Authors: Hamad Alrbyawi

Therapeutic glycoproteins, a pivotal class of biopharmaceuticals, have transformed modern medicine through their broad applications in oncology, immunotherapy, and infectious disease management. Their structural complexity and biological specificity make them highly effective in targeting disease pathways; however, challenges related to stability, bioavailability, and delivery efficacy limit their full potential. Recent advancements in delivery technologies have sought to address these challenges through innovative approaches such as nanotechnology-based carriers, controlled-release systems, and molecular engineering. These strategies have demonstrated the ability to enhance glycoprotein stability, optimize pharmacokinetics, and achieve targeted delivery with minimal off-target effects. This review provides a comprehensive overview of state-of-the-art lipid-based delivery systems specifically designed to overcome the unique pharmaceutical challenges associated with therapeutic glycoproteins, highlighting their design principles, formulation strategies, mechanisms of encapsulation and release, and therapeutic advantages in improving glycoprotein stability, bioavailability, targeted delivery, and treatment efficacy. In addition to surveying the current landscape, this review delves into the key challenges impeding the widespread adoption of advanced delivery systems, including immunogenicity, manufacturing scalability, and clinical translation. The review concludes with insights into emerging trends in the development of lipid-based delivery systems, positioning glycoprotein therapeutics at the forefront of innovation in biopharmaceuticals. This overview of advancements and challenges aims to provide a roadmap for future progress in the field of glycoprotein delivery and therapeutic applications.

Pharmaceutics doi: 10.3390/pharmaceutics18081044

Authors: Manickam Rajkumar Nadarajan Prathap Vivekanand Ankush Kashid Bhupendra G. Prajapati Kokila Palani Parappurath Narayanan Sudha Prabhakaran Rajkumar Biswajit Basu

Mesoporous silica nanoparticles (MSNs) have emerged as versatile nanocarriers for biomedical applications because of their unique physicochemical properties, including high surface area, large pore volume, excellent drug-loading capacity, controllable biodegradation, and facile surface functionalization. These characteristics have enabled the development of advanced drug delivery systems with enhanced therapeutic efficacy, targeted delivery, improved bioavailability, and reduced systemic toxicity. Recent advances in MSN synthesis, physicochemical properties, surface engineering, and functionalization strategies have significantly improved their biological performance and therapeutic potential. In particular, integrating polymers, lipids, and liposomes with MSN platforms has enhanced colloidal stability, circulation time, cellular uptake, and target specificity, thereby facilitating efficient, stimuli-responsive drug delivery. This review highlights MSN-based drug delivery systems in cancer therapy, where multifunctional nanocarriers enable site-specific delivery, controlled drug release, enhanced tumor accumulation, and reduced off-target effects. The review discusses the expanding roles of MSNs in antimicrobial therapy, wound healing, tissue engineering, and regenerative medicine, emphasizing their ability to promote localized therapeutic delivery, immunomodulation, angiogenesis, and tissue regeneration. The review discusses the diagnostic and theragnostic capabilities of MSNs for disease imaging and monitoring. It also critically evaluates current challenges related to biocompatibility, biodegradation, toxicity, biological barriers, large-scale manufacturing, clinical translation, and regulatory considerations. This review provides a comprehensive overview of recent progress, current limitations, and future opportunities for MSN-based platforms in targeted drug delivery and advanced biomedical applications, supporting their continued advancement toward clinical translation and precision medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18081043

Authors: Adila Nazli Bernadett Szögi-Tatár Szilvia Bősze Gergő Tóth Katalin Solymosi Szabolcs Béni Imre Boldizsár

Background: Rosmarinic acid (RA) is a major phenylpropanoid constituent of many medicinal plants belonging to the Nepetoideae subfamily of the Lamiaceae family and contributes significantly to their biological activities. Traditionally, RA-containing extracts have been prepared using organic solvents or hot-water extraction. Methods: In addition to conventional extraction techniques, the phenylpropanoid composition of medicinally important tissues from 13 Nepetoideae species was investigated following incubation in an aqueous medium at room temperature. Phenylpropanoids in the differently prepared extracts were identified by extensive HPLC-UV-HR-MS/MS and NMR analyses. The antioxidant and cytostatic activities of isolated phenylpropanoids were evaluated using DPPH and Alamar Blue assays, respectively. Results: In tissues of all investigated species, RA underwent rapid enzyme-catalyzed conversion during aqueous incubation through oxidative decarboxylation, accompanied by the transient accumulation of nepetoidins A and B. The RA-related phenylpropanoid salvianolic acid K was identified in the roots of three Salvia species, while a previously undescribed isomer, designated salvianolic acid O, was discovered in the roots of Salvia verticillata. Both compounds underwent enzymatic transformations analogous to those of RA in aqueous medium, yielding two previously undescribed phenylpropanoids, salvianolic acids P and Q. Optimized aqueous incubation conditions enabled the high-yield accumulation of these conversion products. Together with their parent compounds, seven phenylpropanoids were isolated from selected plant tissues and were found to exhibit structure-dependent antioxidant and metabolic inhibitory activities in vitro. Conclusions: Aqueous incubation of Nepetoideae plant tissues markedly alters their phytochemical composition and provides access to previously unavailable bioactive phenylpropanoids.

Pharmaceutics doi: 10.3390/pharmaceutics18081042

Authors: Mebarka Ouassaf Bader Y. Alhatlani

Background/Objectives: Pharmacokinetic and pharmacodynamic drug–drug interactions are major determinants of drug safety in polypharmacy, with potential consequences including reduced therapeutic efficacy, altered drug exposure, and increased adverse effects. This study presents DDI-HierPred, an artificial intelligence-based hierarchical framework for drug–drug interaction prediction using Morgan fingerprint-based drug-pair representations. Methods: At Level 1, Logistic Regression, Random Forest, Linear SVM, and XGBoost were compared for pharmacokinetic/pharmacodynamic (PK/PD) classification. At Level 2, an XGBoost multiclass classifier was used to predict 61 interaction subtype classes. End-to-end performance was evaluated using predicted Level 1 routing, and additional drug-identity-disjoint evaluations were conducted to assess generalization when one or both drugs were unseen during training. Y-randomization analyses were performed for both classification levels. Results: Linear SVM achieved the best Level 1 performance, yielding an accuracy of 0.8661, a ROC-AUC of 0.9380, and an MCC of 0.7303 on the independent test set. At Level 2, the XGBoost classifier achieved an accuracy of 0.8712, a balanced accuracy of 0.9087, a macro F1-score of 0.9115, and a Top-3 accuracy of 0.9868. Because the Level 2 test partition was also used for algorithm comparison and model selection, these results should be regarded as exploratory and potentially optimistic rather than as an independent final evaluation. When evaluated end-to-end using predicted Level 1 routing, performance decreased to an accuracy of 0.5767, balanced accuracy of 0.3573, and macro F1-score of 0.4379. Additional drug-identity-disjoint evaluations showed further performance reductions when one or both drugs were unseen during training, highlighting the greater difficulty of generalization to previously unseen drug identities. Y-randomization analyses supported the robustness of both classification levels. Conclusions: The framework was deployed as a publicly accessible web platform integrating documented interaction lookup, hierarchical PK/PD classification, Level 1-constrained subtype prediction, confidence scoring, single-pair and batch analysis, ranked Top-3 predictions, and downloadable reports. These findings indicate that upstream routing and unseen-drug generalization remain important limitations of the current framework. DDI-HierPred therefore provides a computational platform for research-oriented screening, interpretation, and prioritization of potential drug–drug interactions.

Pharmaceutics doi: 10.3390/pharmaceutics18081041

Authors: Hua Jiang Zhong-Kang Yang Jun Li Yu-Pin Wang

Background/Objectives: Co-amorphous solid dispersions (c-ASD) are a promising strategy for enhancing the dissolution of poorly water-soluble drugs. Naringin (NA) and neohesperidin (NE) are dihydroflavonoid components and are poorly soluble. They can form a c-ASD. Concerning the c-ASD formation mechanism, the intermolecular forces between the non-sugar aglycones of NA and NE have been determined; however, the roles of the sugar chains, which account for nearly 50% of the overall molecular weight of both compounds, are unclear. Therefore, the impact of the sugar moiety on the solubility of NA-NE c-ASD needs to be investigated. Methods: The sugar removal products of NA and NE are naringenin-7-O-glucoside, naringenin, hesperetin-7-O-glucoside, and hesperetin. Therefore, in this study, the solubility profiles of NA with hesperetin-7-O-glucoside and hesperetin, and of NE with naringenin-7-O-glucoside and naringenin were assessed by dissolution determination and analyzed by PXRD. Results: These results indicated that the rhamnose and glucose moieties on the NE sugar chain and the glucose moiety on the NA sugar chain are vital to the stability and solubility of NE-NA c-ASD. The rhamnose moiety on the sugar chain of NA is removable, without which a stable soluble aggregator may also be constructed, but the soluble aggregator formation ability is weakened. Conclusions: The results of this study not only inform the rational selection of co-formers for structurally similar flavanone glycosides but can also help chemists modify the c-ASD structure based on the sugar moiety.

Pharmaceutics doi: 10.3390/pharmaceutics18081040

Authors: Xavier Buin Rosy Ghanem Ines Pankonien Frédéric Becq Margarida Amaral Tristan Montier

Since the development of the first cystic fibrosis transmembrane conductance regulator (CFTR) modulator in 2012, these therapies have revolutionized patients’ health. They are now the most effective treatment for people with cystic fibrosis (pwCF). In fact, elexacaftor/tezacaftor/ivacaftor and vanzacaftor/tezacaftor/deutivacaftor, the latest combination therapies consisting of a CFTR potentiator and two CFTR correctors, improved lung function by 14% in pwCF. Other modulator therapies targeting CFTR mRNA and/or protein are currently under preclinical/clinical investigation. However, due to the variant-specific nature of these therapies, about 10% of pwCF in Europe remains without effective treatment, and many treated pwCF experience various adverse events such as headaches, infections, hepatotoxicity, hypertension, and depression. Therefore, mutation-agnostic strategies such as gene therapy are needed. They could expand treatment eligibility for all pwCF and improve outcomes. In fact, nucleic acid delivery (e.g., pDNA, mRNA, oligonucleotides, genome editing) or targeting non-CFTR channels to restore ion transport represent promising future additional directions for CF therapy. This review aims to discuss a potential combination between gene therapy approaches and existing modulators to improve treatment eligibility, safety, and efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18081039

Authors: Anna Cargnoni Serafina Farigu Pietro Romele Andrea Papait Marta Magatti Antonietta Silini Ornella Parolini

Background/Objectives: Mesenchymal stromal cells from the amniotic membrane (hAMSCs) counteract fibrosis progression, primarily via anti-inflammatory effects like promoting macrophage polarization toward an anti-inflammatory, pro-regenerative phenotype. Methods: We investigated hAMSCs’ ability to prevent and halt lung fibrosis in a bleomycin-induced fibrosis murine model. We focused on their impact on recruitment and polarization of different macrophage populations, including SPARC- and CD169-expressing macrophages, implicated in resolving pulmonary inflammation and fibrosis. Results: hAMSCs, administered early (concomitant with bleomycin, during acute inflammation), or late (at day 7 post-bleomycin, during established fibrosis), showed anti-fibrotic activity, preserving alveolar area, reducing the extent of lung fibrosis, and decreasing α-SMA levels. These preventive and late anti-fibrotic effects of hAMSCs are associated with a context-dependent presence of distinct macrophage signatures. Early treatment reduced macrophage recruitment and increased levels of Arg1+/iNOS− macrophages, curbing injury-induced inflammation. Late treatment uniquely increased the lung levels of CD169+ macrophages, suggesting their contribution to hAMSCs’ anti-fibrotic effect. We hypothesized a potential involvement of lung CD169+ macrophages in promoting recruitment of regulatory T cells (Tregs) to the lungs. Although these macrophages can establish a CCL22-CCR4 axis with Tregs, and treatment effectively boosted Treg lung levels, the Treg increase is not directly attributable to higher CD169+ macrophage numbers, implying other potentially IL-10-driven mechanisms. Conclusions: hAMSC treatment effectively prevents and blocks lung fibrosis. These effects are associated with distinct lung macrophage marker profiles, suggesting a potential involvement of different macrophage populations in a time-dependent manner; thus highlighting administration timing’s role in optimizing therapeutic synergy.

Pharmaceutics doi: 10.3390/pharmaceutics18081038

Authors: Mohamed Aghyad Al Kabbani Laura Köhler Tamara Wied Daniel Adam Jennifer Klimek Hans Zempel

Background: Microtubules play a crucial role in neuronal structure and function, with their stability and dynamics regulated by posttranslational modifications (PTMs) such as polyglutamylation. In Alzheimer’s disease (AD), the microtubule-associated protein TAU becomes mislocalized into the somatodendritic compartment (‘TAU missorting’), dissociates from microtubules, aggregates into neurofibrillary tangles, and contributes to microtubule destabilization and neuronal death. Objectives and Methods: Here, we investigated the role of tubulin tyrosine ligase-like proteins (TTLLs) in TAU missorting and microtubule dysregulation using human-induced pluripotent stem cell (hiPSC)-derived cortical neurons treated with oligomeric amyloid-beta (oAβ) to replicate AD-like conditions. TTLL1, TTLL4, and TTLL6 were selectively knocked down (KD) to assess their impact on TAU missorting and microtubule stability. Fluorescence resonance energy transfer (FRET) microscopy was used to examine proximities between TAU and TTLL proteins. Results: We observed TAU missorting, increased tubulin polyglutamylation, decreased tubulin acetylation associated with microtubule destabilization, and synaptic declustering in oAβ-treated neurons. TTLL1 KD significantly reduced TAU missorting, tubulin polyglutamylation, and synaptic disintegration, while TTLL4 KD showed moderate effects, and TTLL6 KD restored microtubule acetylation. Importantly, TTLL KD did not impair neuritic networks, dendritic complexity, or neuronal activity. FRET microscopy in HEK293T cells revealed a close molecular proximity between TAU and TTLL1 consistent with a potential direct or complex-mediated association, but not with other TTLLs, suggesting a direct role of TTLL1 in TAU-mediated toxicity. Conclusions: Our findings identify TTLL1 as a promising therapeutic target for limiting TAU-associated cytoskeletal pathology in AD. These results support further development of pharmacological or genetic strategies targeting TTLL1 as a disease-modifying approach for AD and related tauopathies.

Pharmaceutics doi: 10.3390/pharmaceutics18081037

Authors: Yaqian Gao Yiheng Liang Haiyan Chen Zigui Wang Yanchang Huang Wei Zhou Zhiyun Du

Background: Adequate inner-ear exposure is a key challenge in treating age-related hearing loss (ARHL). Astragaloside IV (AS-IV), the principal saponin of Astragalus membranaceus, has anti-aging activity but poor oral bioavailability; its aglycone cycloastragenol (CAG) may represent a pharmacokinetically optimized active moiety. Methods: We compared the pharmacokinetics, cochlear exposure, and biotransformation of CAG and AS-IV after oral dosing in mice by LC-MS/MS, and evaluated CAG in a D-galactose-induced accelerated-aging rat model (ABR, hair-cell morphology, redox and cytokine assays) and in D-galactose-stressed HEI-OC1 cells. Results: CAG achieved markedly higher plasma and perfused whole-cochlea exposure than intact AS-IV after equivalent dosing, even after molar-dose normalization. AS-IV was biotransformed to CAG in vivo and in liver microsomes, supporting CAG as a quantitatively important active metabolite. In rats, oral CAG reduced ABR threshold elevation and preserved hair-cell architecture, improved cochlear redox status (higher SOD/GSH; lower ROS/MDA), and lowered serum TNF-α/IL-6. In HEI-OC1 cells, CAG attenuated mitochondrial membrane-potential loss and apoptosis. Conclusions: At equimolar concentrations, CAG and AS-IV showed comparable protective activity in vitro, indicating that the advantage of CAG resides mainly in its pharmacokinetic profile. CAG thus attenuates auditory dysfunction in an accelerated-aging model with superior exposure, supporting its further development for age-associated hearing disorders.

Pharmaceutics doi: 10.3390/pharmaceutics18081036

Authors: Yu Zhang Jian Guo Haonan Qiu Jiale Liu Chi Zhang Lutan Zhou Chunfei Wang Lihua Li Xuefeng Hou

Background: Imperatorin (IPT) is a natural furanocoumarin featuring robust anti-inflammatory, antifibrotic and antioxidant activities. However, poor aqueous solubility and insufficient oral bioavailability restrict its clinical application. Multiple gastrointestinal barriers, including the mucus barrier, limited epithelial penetration and P-glycoprotein-triggered drug efflux, are major obstacles hindering IPT oral absorption. Methods: N-acetylcysteine (NAC)-functionalized TPGS conjugates were synthesized first. Using Pluronic® F108 and Lipoid® S-100 as a matrix, imperatorin@N-acetylcysteine-TPGS/Pluronic® F108/Lipoid® S-100 (IPT@NAC-TFS) micelles were fabricated. We characterized their physicochemical features and in vitro release behavior. The Caco-2/HT29-MTX-E12 co-culture cell model was adopted to explore mucus permeation, cellular uptake and transepithelial transport mechanisms. In vivo intestinal distribution and pharmacokinetic tests in rats were carried out to confirm the oral absorption-enhancing effect of micelles. Results: Optimized micelles displayed a uniform shape and favorable encapsulation efficiency. Low CMC maintained structural stability upon gastrointestinal dilution. NAC modification conferred mucus-penetrating capacity on micelles. TPGS simultaneously improved epithelial barrier permeability and inhibited drug efflux, switching IPT transport mode. The micelles effectively cleared intracellular ROS, recovered SOD activity and lowered MDA levels in BLM-impaired MLg fibroblasts. In vivo results revealed enhanced intestinal drug accumulation, with the relative oral bioavailability of IPT increased by 6.07-fold. Conclusions: IPT@NAC-TFS micelles overcome multiple gastrointestinal barriers for oral IPT delivery. Combining mucus penetration, efflux suppression and antioxidative capacity, this system offers a promising strategy to develop oral formulations of poorly soluble antifibrotic natural products.

Pharmaceutics doi: 10.3390/pharmaceutics18081035

Authors: Emilija Nemickaite Pooja Todke Vaidotas Cicenas Elena Jasiūnienė Mindaugas Marksa Jurga Bernatoniene

Background: Three-dimensional printing (3DP) is rapidly advancing personalised medicine, yet systematic performance comparison with conventional manufacturing remains limited, particularly for plant-based formulations. Methods: This study compared tablets containing plant-based APIs (cannabidiol, apigenin, and luteolin) produced via conventional hot moulding and semi-solid extrusion (SSE) 3DP. The formulations were evaluated for physicochemical, mechanical, rheological, structural, and drug-release properties. Results: Both manufacturing methods produced tablets with comparable dimensions and mass; however, pronounced formulation-dependent differences were observed in mechanical strength, rheology, and microstructure. The molecular modelling predictions were consistent with the experimental findings. Agar–pectin exhibited the strongest predicted polymer–polymer and polymer–API interactions, including multiple hydrogen bonds, and formed a comparatively dense and cohesive matrix associated with slower API release. In contrast, the weaker interactions predicted for gelatine–pectin were associated with a less cohesive and more porous matrix that facilitated medium penetration, API diffusion, and drug release. SSE printlets generally exhibited greater porosity and more heterogeneous internal architectures than moulded tablets, resulting in enhanced drug release of approximately 95%. Micro-CT analysis provided important structural confirmation; API incorporation increased the void volume of gelatine–pectin printlets from 1.15% to 8.77%, demonstrating that disruption of polymer interactions contributed to pore formation and enhanced molecular diffusion. The observed release behaviour correlated with predicted molecular interactions and experimentally observed microstructural features, where increased porosity and weaker polymer–API interactions facilitated enhanced drug diffusion. Conclusions: Overall, SSE-3DP outperformed conventional moulding, demonstrating superior tunability and performance. This work provides a mechanistically informed strategy for designing plant-based, personalised natural products using 3DP technologies.

Pharmaceutics doi: 10.3390/pharmaceutics18081034

Authors: Gorana Ilić Ivana Beara Ljiljana Milovanović Aleksandra Jovanović Dragana Dekanski Andrea Pirković

Background/Objectives: Vitis vinifera is a rich source of polyphenolic compounds that are known for their benefits on skin health, including antioxidant, anti-inflammatory, and photoprotective effects. Red wines have shown a positive impact on cardiovascular disease, diabetes, and cancer prevention; however, evidence regarding their effects on skin biology remains scarce. This study evaluated red wine concentrates obtained from Cabernet Sauvignon and Merlot varieties, selected based on their previously demonstrated biological activities, to identify the samples with the highest potential for skin health applications by assessing their inhibitory activity against skin-related enzymes (tyrosinase, elastase, and collagenase), along with their effects on HaCaT cells, including cytotoxic, antioxidant, anti-inflammatory, and wound healing activities. Methods: Enzyme inhibition studies were based on spectrophotometric methods; the MTT and CV assays were carried out to test the cytotoxicity, and the DCFH-DA assay was employed to evaluate the antioxidant activity of the wine concentrates; wound healing potential was examined by the cell scratch assay, and anti-inflammatory activity was investigated using ELISA and qPCR analyses. PCA of the samples and their biological activity was performed as well. Results: Examined red wine concentrates exhibited significant enzyme inhibitory activity and showed no cytotoxicity on HaCaT cells up to the concentration of 50 μg/mL; the samples stimulated cell migration, inhibited intracellular ROS production in AAPH-stressed keratinocytes, and selectively modulated TNF-α-induced inflammatory responses, reducing IL-8 secretion while enhancing certain cytokine transcripts, underscoring the complexity of their bioactive effects. Conclusions: Red wine concentrates showed significant potential as skin-beneficial agents, demonstrating strong antiaging, marked cellular antioxidant effects and wound healing activity when applied at non-cytotoxic concentrations. Multivariate analysis revealed distinct bioactivity profiles among the wine concentrates.

Pharmaceutics doi: 10.3390/pharmaceutics18081033

Authors: Filipe Figueiredo Kristian Gillebo Sørmo Aleksander Pettersen Heng Chi Jaya Kumari Swain Toni Erkinharju Trilochan Swain Shripathi Bhat Kim Præbel Stefano Peruzzi Jacques Godfroid Øystein Evensen Hetron Mweemba Munang’andu Roy Ambli Dalmo

Background: Pancreas disease, caused by salmonid alphavirus subtype 3 (SAV3), remains a significant threat to Atlantic salmon (Salmo salar L.) aquaculture in Norway despite the widespread use of inactivated vaccines. Molecular adjuvants represent a promising strategy to improve vaccine efficacy by enhancing immune responses. Methods: In this study, we evaluated the immunological and protective effects of two transcription factors, gata3 and eomes encoded by plasmid DNA adjuvants in combination with a low-dose inactivated SAV3 vaccine. Atlantic salmon were immunized with inactivated virus alone or in combination with either gata3- or eomes-encoding plasmids, followed by SAV3 challenge. Vaccine performance was assessed by growth, antibody response (ELISA and virus neutralization assay), and histopathological scoring of heart and pancreas. Results: The effects of gata3 and eomes were context-dependent: eomes reduced pancreas lesions at 4 weeks post-challenge, while both plasmid-adjuvant groups showed improved growth at 10 weeks post-challenge compared to the plasmid control. Although antibody responses were increased in all vaccinated groups at 6 weeks post-immunization, the plasmid control performed comparably to the GATA3 or EOMES groups, and no significant differences were detected in the virus neutralization test. Interestingly, the empty plasmids given to control fish exhibited immunostimulatory activity, warranting further investigation. In vitro and in vivo fluorescence microscopy confirmed expression of the transcription factors in transfected cells and at the injection site. Conclusions: Our results suggest that transcription factor-encoding plasmids can provide selective adjuvant effects, while also highlighting the potential that plasmid DNA can have as adjuvants in fish vaccine development.

Pharmaceutics doi: 10.3390/pharmaceutics18081032

Authors: Lee Ann Applegate Farid Hadjab Sandra Jaccoud Alexandre Porcello Virginie Philippe Nathalie Hirt-Burri Corinne Scaletta Brigitte M. Jolles Dominique P. Pioletti Robin Martin Alexis E. Laurent

Background: The clinical translation of cell-based therapies for knee articular cartilage repair is fundamentally restricted by the severe biological unpredictability of autologous cell sources, inherent manufacturing bottlenecks, and the rapid phenotypic dedifferentiation of cells expanded in conventional 2D monolayers. To overcome these translational hurdles, this study engineered a scaffold-free, 3D formulation of highly characterized allogeneic FE002-Cart chondroprogenitor spheroids. Methods: We systematically investigated the specific microenvironmental cues and Good Manufacturing Practice (GMP) formulation parameters required to direct functional chondrogenesis. The structural and biochemical performance of this allogeneic formulation was benchmarked against multiple primary adult autologous chondrocyte types. Finally, we evaluated the phenotypic resilience of the microtissues in simulated osteoarthritic (OA) environments and investigated both short-term liquid storage and advanced terminal preservation strategies to establish off-the-shelf logistical viability. Results: Precise microenvironmental regulation proved to be a critical biological prerequisite. The synergistic combination of physiological hypoxia (2% O2) and stringent glucocorticoid limitation (10 nM dexamethasone) induced robust glycosaminoglycan (GAG) deposition and a > 200-fold upregulation of ACAN and COL2, while suppressing the terminal hypertrophic drift observed in adult chondrocytes. Benchmarking revealed that the allogeneic FE002-Cart formulation substantially mitigates the profound morphological and biochemical unpredictability inherent to adult autologous cell sources. Furthermore, the scaffold-free spheroid geometry yielded a 10-fold increase in GAG production per cell compared to traditional matrix-seeded (MACI) platforms. Transitioning to a GMP-compatible manufacturing process revealed extreme cellular sensitivities; excipients within standard pharmaceutical-grade dexamethasone severely aborted chondrogenic differentiation, emphasizing the necessity of rigorous raw-material qualification. Functionally, the 3D architecture acted as a protective physical shield, sustaining high cellular viability when subjected to severe inflammatory stress and 100% OA patient synovial fluid. Logistically, the viable spheroids maintained matrix integrity and inter-spheroid fusion potential for up to 7 days at ambient temperature in transport medium. Finally, advanced spheroid preservation via lyophilization and high-dose gamma irradiation eliminated biological viability but successfully transitioned the microtissues into highly organized, terminally irradiated matrices capable of heterologous in vitro structural merging. Conclusions: These findings define the critical biological thresholds for manufacturing, demonstrate the enhanced in vitro biosynthetic efficiency of 3D allogeneic microtissues compared to specific autologous and matrix-dependent baselines, and establish a highly practical, off-the-shelf logistical framework for the regenerative treatment of large knee chondral defects.

Pharmaceutics doi: 10.3390/pharmaceutics18081029

Authors: Sarah Hunachagi Hoor Hashim Alqudihi Sayed AbdulAzeez J. Francis Borgio Dana Almohazey

Background/Objectives: Colorectal cancer (CRC) remains a leading cause of cancer-associated mortality worldwide. The current therapeutic interventions are heavily constrained by the development of resistance and severe systemic toxicity. To address these challenges, this study integrated a multi-disciplinary framework involving high-throughput in silico screening followed by in vitro experimental validation to identify novel genetic targets of CRC and evaluate the efficacy of FDA-approved drugs. The primary objective was to identify safe and selective therapeutic agents capable of modulating their effect. Methods: The methodology employed a systematic screening of recent large-scale Genome-Wide Association Studies (GWASs) to pinpoint novel targets, followed by in silico pathogenicity prediction, homology modelling and high-throughput virtual screening of over 1615 FDA-approved drugs. The prioritized candidates were validated in vitro using MTT cytotoxicity assays and differential gene expression analysis across CRC cell lines (HCT116 and HT29) and a non-tumorigenic control, Human embryonic kidney cell line HEK293. Results: In silico analysis identified CLUH, CLSTN3 and SLC11A2 as novel potential targets. Based on in silico predicted deleterious mutations and subsequent molecular docking-based virtual screening, Telmisartan, Dutasteride and Venetoclax were prioritized. This prioritization was supported by their high binding affinity and dose-dependent cytotoxicity in MTT assays; thus, suggesting their repurposing potential for CRC treatment. Telmisartan exhibited a superior therapeutic profile not only in terms of the statistically significant cytotoxicity (p < 0.01), but also its selective effect on HCT116 and HT29 when compared to high safety profile in HEK293. This was further validated when Telmisartan selectively downregulated CLUH and SLC11A2 in CRC cell lines, HCT116 and HT29 while maintaining expression levels in the non-cancerous HEK293 cell line remained significantly unaffected. Furthermore, a 100 ns molecular dynamics simulation confirmed the stable binding conformation and structural reliability of the SLC11A2 (Trp179Ser)–Telmisartan complex. Conclucions: Our findings conclude that Telmisartan is a promising candidate for drug repurposing for CRC treatment and capable of modulating selected novel biomarkers CLUH and SLC11A2. However, further multi-omics-based confirmatory studies and pre-clinical validation studies are needed in the future to confirm the long-term efficacy of this repositioning strategy.

Pharmaceutics doi: 10.3390/pharmaceutics18081030

Authors: Mengmeng Liu Guoqing Liu Qixian Ling Yongbo Chen Haojie Xu Runhui Wu Zhenping Chen Libo Zhao

Background: Prophylaxis in children with hemophilia B (HB) lacks quantitative approaches that integrate both plasma exposure and tissue distribution. This study aimed to develop and validate a physiologically based pharmacokinetic (PBPK) model of factor IX (FIX) for pediatric HB. The model incorporated the binding of FIX to type IV collagen (Col4) to characterize its distribution in both plasma and extravascular tissues. Methods: A total of 20 children with severe HB were included, contributing 219 plasma samples. The base PBPK model was first established and verified using adult and plasma-derived FIX (pdFIX) data. It was subsequently extrapolated to children by integrating FIX-CTBB parameters and pediatric observations for model calibration. The validated model was used to characterize plasma pharmacokinetics, predict tissue distribution and target attainment, and simulate alternative prophylactic dosing regimens. Results: The model adequately described the plasma pharmacokinetics of FIX in children and predicted substantial extravascular distribution. Total extravascular exposure was approximately sixfold higher than plasma exposure. Marked heterogeneity in target attainment was identified across tissues. Lower target attainment was observed in the colon, pancreas, and brain, whereas delayed attainment occurred in bone and muscle. Simulations of prophylactic dosing regimens suggested that 75 IU/kg twice weekly may provide a favorable balance among sustained FIX exposure, tissue-level target attainment, and treatment burden. Conclusions: This PBPK model provides a mechanistic and quantitative framework for characterizing plasma and tissue exposure to FIX in children with HB and may support individualized optimization of FIX prophylactic dosing.

Pharmaceutics doi: 10.3390/pharmaceutics18081031

Authors: Yuchen Qu Zhuan Yang Wen Ma Peng Xiao Yani Gu Jie Pan Xinyun Zhang Chen Zhao Yunli Yu

Background: In patients with atrial fibrillation receiving long-term anticoagulation therapy with dabigatran etexilate (DABE), moxifloxacin (MFLX) is frequently coadministered to treat concurrent infections; however, the potential drug–drug interaction (DDI) between these agents remains unclear. Herein, we examined the underlying mechanism by which MFLX attenuates DABE pharmacokinetics in rats; subsequently, we elucidated the DDI in humans by establishing a physiologically based pharmacokinetic (PBPK) model based on these animal data. Methods: The 3- and 14-day effects of 40 mg/kg MFLX once daily and secondary bile acid (SBA)-containing dietary intervention on the pharmacokinetic profile of DABE and its active form, dabigatran (DAB), were examined in a rat model. Ileum tissues were harvested to measure the expression of P-glycoprotein (P-gp), pregnane X receptor (PXR), and peroxisome proliferator-activated receptor alpha (PPARα). In addition, we examined the effects of secondary bile acids (SBAs) on P-gp expression and quantified P-gp-mediated DABE efflux transport activity in Caco-2 cells. A PBPK model was used to predict the risk of DAB exposure under this DDI scenario and under combined high-risk conditions, including renal impairment and advanced age. Results: Treatment with MFLX for 3 and 14 days inhibited SBA-producing gut microbiota, thereby suppressing the conversion of primary bile acids to SBAs. Concurrently, a marked reduction in intestinal P-gp expression was observed, along with a significant enhancement of the oral bioavailability of DABE. These effects were reversed by SBA-containing diets. In vitro experiments using Caco-2 cells revealed that physiologically relevant concentrations of SBA significantly upregulated P-gp expression and function, whereas MFLX incubation alone showed no direct modulatory effect on these transporters or regulators. PBPK simulation results showed that in vivo exposure of DAB would increase by 41.7%, 104%, 251%, and 115% when coadministered with MFLX alone, with coexisting mild renal impairment, moderate renal impairment, and aging, respectively. Conclusions: MFLX increases DAB exposure by reducing SBA-regulated intestinal P-gp function. PBPK simulations suggest a low risk of DDI from MFLX coadministration alone; however, caution is warranted in patients with aging or renal impairment.

Pharmaceutics doi: 10.3390/pharmaceutics18081028

Authors: Yi Su Yingxin Lu Jiahui Yu Chen Jin Yi Chen Wei Lu Jian Ding

Background and Objective: Pancreatic ductal adenocarcinoma (PDAC) is characterized by dense extracellular matrix (ECM) with overexpression of extra domain B fibronectin (EDB-FN). Methods: Herein, we investigated novel albumin-binding peptide–drug conjugates, Mc-ZD2-SN38 and SSC-ZD2-SN38, which integrate the prolonged circulation properties of albumin with the active targeting capability of the ZD2 peptide toward EDB-FN. Covalent and noncovalent albumin-binding strategies extended the half-life by approximately 100-fold and 15-fold, respectively, compared with the non-albumin-binding peptide–drug conjugate (PDC) (ZD2-SN38). Results: Notably, ZD2-mediated targeting markedly reduced payload accumulation in major organs, resulting in an improved safety profile. In a BxPC-3 xenograft model, Mc-ZD2-SN38 and SSC-ZD2-SN38 demonstrated potent and sustained antitumor efficacy. Conclusions: These findings establish a long circulation and active targeting drug delivery strategy, providing an ideal framework for the development of PDCs with enhanced therapeutic indices.

Pharmaceutics doi: 10.3390/pharmaceutics18081027

Authors: Lomass Soliman Dóra Paróczai Katalin Burián Rita Ambrus

Background/Objectives: The stability and biological safety of newly developed formulations must be established to support their therapeutic efficacy and clinical translation in pulmonary drug delivery. Therefore, this follow-up study comprehensively evaluated the short- and long-term stability and the in vitro cytotoxicity of optimized, co-spray-dried theophylline (THN) dry powders for inhalation against A549 lung epithelial cells. Methods: Two established formulations were selected: THN-RAF (raffinose–leucine–glycine based) and THN-TRE (trehalose–leucine based). Stability was assessed under accelerated conditions (40 °C/75% RH, 3 months) and long-term desiccator storage (25 °C, 1 year) using laser diffraction, SEM, XRPD, FTIR, DSC, TGA, and Andersen Cascade Impaction. As THN-TRE had been previously confirmed to be cytocompatible, only THN-RAF and its components were evaluated against A549 human alveolar epithelial cells using the MTT assay. Results: Under accelerated conditions, both formulations exhibited pronounced recrystallization (Xc up to 89.9%), agglomeration (D [0.9] up to 217.08 µm for THN-TRE), and deterioration in aerodynamic performance (FPF as low as 11.55%, MMAD up to 6.68 µm). By contrast, long-term desiccator storage induced substantial recrystallization (Xc up to 80.7%) while preserving thermal, chemical, and aerodynamic performance (FPF ≈ 40%; MMAD 4.99–5.21 µm). THN-RAF was more resistant to stress-induced agglomeration than THN-TRE. Cytotoxicity assessment confirmed cytocompatibility of THN-RAF, with cell viability exceeding 70.99% at all tested concentrations (up to 500 µg/mL). Conclusions: These findings reveal a marked discrepancy between the outcomes of ICH accelerated testing and long-term desiccator storage. They underscore the importance of considering moisture-protective packaging configurations when designing stability protocols for amorphous inhalable formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18081026

Authors: Harshada B. Pawar Pawan Ganesh Nayak Amatha Sreedevi Ramya Ravi Pradeep M. Muragundi

Advancements in pulmonary drug delivery have enabled effective treatment approaches for more severe disease conditions, such as chronic obstructive pulmonary diseases, asthma, cystic fibrosis, and other pulmonary disorders, via targeted, sustained, and immediate drug delivery routes with minimal systemic side effects. However, conventional delivery systems have many limitations, such as poor drug targeting, adherence, and deposition, which ultimately cause variations in drug profiles and therapeutic efficacy. Recent advances in artificial intelligence (AI) and machine learning (ML) have enabled the development of smart inhaler drug–device combination systems for personalized therapy using predictive formulation parameters, design variables, device performance, and inhalation pattern monitoring. Advanced AI techniques, such as artificial neural networks, deep learning, random forests, support vector machines, deep learning algorithms, and computational modeling, predict the mass median aerodynamic diameter (MMAD), fine-particle fraction (FPF), emitted dose, and regional lung deposition. Smart inhalation devices coupled with digital sensors and computing systems enable the real-time monitoring of inhalation profiles and adherence. Moreover, AI- and ML-enabled Quality by Design (QbD) and digital twin framework technologies enhance the optimization of manufacturing process parameters, consistency, robustness, and scale-up performance. Although several developments have been reported, there is still room for improvement in terms of data heterogeneity, algorithm transparency, interpretability, cybersecurity, regulations, and long-term clinical standardization. This review emphasizes the use of AI to improve the performance of pulmonary drug delivery through smart inhaler drug–device combination therapies, focusing on technological advancements, formulation optimizations, smart inhalers, regulatory issues, current limitations, and future perspectives of AI-based pulmonary drug delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18081025

Authors: Qing Sun Yang Fu Tianwei Wang Zongyuan Xu Zeping Gui Kun Liu Xuzhong Liu

Background/Objectives: Renal ischemia–reperfusion injury (IRI) and infection-associated renal damage are characterized by persistent inflammation, oxidative stress, microvascular dysfunction, and impaired tissue regeneration, creating a hostile microenvironment that limits effective repair. We developed an injectable, photocrosslinkable methacrylated chitosan (CSMA) hydrogel for the localized co-delivery of cerium oxide nanoparticles (CeO2NPs) and vascular endothelial growth factor (VEGF), aiming to integrate redox modulation, antibacterial activity, and regenerative support. Methods: Gelation, microstructure, rheology, degradation, and CeO2NP/VEGF were characterized. Tubular epithelial and fibroblast migration and endothelial network formation, angiogenic gene expression, and antibacterial activity against Staphylococcus aureus and Escherichia coli were evaluated in vitro. Theraputic performance was assessed by renal surface application in a rat renal IRI model and catheter-mediated interavsical administration in an ascending urinary tract infection model. Systematic biocompatibility was evaluated separately in a 14-day subcutaneous implantation study. Renal response were further investigated using transcriptomic and targeted molecular analyses. Results: The CSMA/VEGF/CeO2NPs hydrogel exhibited rapid in situ gelation, interconnected porous architecture, stable viscoelasticity, gradual degradation, and sustained release of both CeO2NPs and VEGF. The formulation enhanced tubular epithelial and fibroblast migration, promoted endothelial network formation and angiogenic gene expression and effectively inhibited both S. aureus and E. coli. In a surgically controlled rat renal IRI model, direct renal-surface application of the hydrogel reduced tubular injury, inflammatory infiltration, and fibrotic remodeling. In a separate ascending urinary tract infection model, catheter-based intravesical administration reduced the ascending renal bacterial burden and infection-associated inflammatory injury. No detectable adverse systemic effects observed under the tested conditions over the 14-day observation period in the subcutaneous implantation. Transcriptomic analyses further revealed that CSMA/VEGF/CeO2NPs treatment was associated with marked remodeling of the renal injury microenvironment, characterized by suppression of antigen presentation and immune activation pathways, alongside restoration of metabolic programs associated with amino acid, lipid, and purine metabolism. These molecular changes were accompanied by downregulation of CIITA/CD74/MHC-II signaling, recovery of metabolic regulators AGXT and ACOX1, modulation of Hippo/YAP- and ECM-associated pathways, and preservation of renal structural markers including nephrin and WT1. Conclusions: The localized CSMA-mediated co-delivery of CSMA/VEGF/CeO2NPs hydrogel promotes renal repair through resolution of maladaptive immune activation, metabolic reprogramming, angiogenic enhancement, and preservation of renal structural integrity, providing a promising biomaterial strategy for the treatment of ischemic and infection-associated renal injuries.

Pharmaceutics doi: 10.3390/pharmaceutics18081024

Authors: Anam Sajjad Khan Daniela Müller Cornelia M. Keck

Background: Lipid nanoparticles are widely investigated as oral drug delivery systems, but their intestinal performance remains difficult to predict based only on physicochemical properties. This study aimed to elucidate how the lipid matrix composition influences the intestinal permeation of curcumin from lipid nanoparticles. Methods: Curcumin-loaded nanoemulsions, nanostructured lipid carriers with defined solid-to-liquid lipid ratios, and solid lipid nanoparticles were prepared by high-pressure homogenization. All formulations were characterized with respect to particle size, polydispersity index, and zeta potential before and after simulated intestinal pre-incubation in a simplified SDS-containing intestinal fluid. Intestinal permeation was evaluated ex vivo using porcine gut tissue by analysis of semi-quantitative fluorescence-based permeation readouts (ART) and mean permeation depth (MPD) after 30 and 60 min. Results: All formulations maintained stable physicochemical properties with particle sizes around 200 nm and negative zeta potentials; pre-incubation increased the negativity of the zeta potential but left particle size unchanged. Despite similar attributes, the formulations differed in intestinal curcumin permeation based on time and composition. At 30 min, nanoemulsions and mixed nanostructured lipid carriers achieved the highest performance. By 60 min, lipid carriers with more liquid lipid significantly increased both the fluorescence intensity and the depth of curcumin permeation, while other systems showed little further improvement. Conclusions: The intestinal permeation of drug from lipid nanoparticles is governed by the lipid matrix architecture and its interaction with the hydrated intestinal environment, which together affect drug-release kinetics and the ability to sustain a trans-epithelial concentration gradient over time. Thus, optimizing oral lipid nanoparticles requires time-resolved, biologically relevant models rather than physicochemical characterization alone, consistent with observed similar matrix-driven effects in dermal delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18081023

Authors: Mirna Perkušić Laura Nižić Nodilo Mario Jug Cvijeta Jakobušić Brala Regina Scherließ Anita Hafner

Background/Objectives: The aim of this study was to develop and apply a novel integrated approach for predicting local mucosal tolerability of spray-dried chitosan/mannitol microspheres previously developed for nose-to-brain donepezil delivery. Methods: Microspheres were prepared by ultrasonic spray-drying, and process reproducibility was evaluated based on particle size distribution, entrapment efficiency, and process yield across independent batches. A lactose-based formulation served as a comparative control. A novel biomimetic model was developed to investigate water evaporation under simulated nasal conditions, enabling prediction of formulation dehydration and crust-like layer formation on the nasal mucosa during nasal residence time. Donepezil-loaded chitosan microspheres and their physical mixture with mannitol were used as controls. Analyses were complemented by solid-state and rheological characterization to elucidate the effects of formulation composition and processing on the observed behavior. Irritation potential was further assessed using the established slug mucosal irritation (SMI) assay. Results: Reproducible microsphere size distribution (Dv10 11.5 ± 1.1 µm, RSD 9.6%; Dv50 28.4 ± 3.9 µm, RSD 13.7; Dv90 61.3 ± 8.4 µm, RSD 8.4%), entrapment efficiency (99.6 ± 1.8%, RSD 1.8%) and process yield (40.9 ± 5.5%, RSD 13.3%) confirmed the robustness of the ultrasonic spray-drying. Replacing mannitol with lactose failed to achieve the desired particle size distribution, highlighting the key role of mannitol under the investigated processing conditions. The biomimetic model coupled with rheological studies demonstrated that chitosan-based gels formed by microsphere swelling in simulated nasal fluid, maintain viscosity, resist dehydration, and undergo rehydration. Additionally, mannitol enhanced water retention and reduced evaporation without increasing occlusivity or the risk of mucosal dehydration. Furthermore, powders containing mannitol exhibited a lower irritation potential in the SMI assay compared to chitosan microspheres alone. Conclusions: Mannitol is a critical determinant of the performance of donepezil-loaded chitosan-based microspheres, contributing to the desired particle size distribution, process reproducibility, favorable hydration and improved mucosal tolerability, thereby supporting the suitability of this platform for nasal donepezil delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18081022

Authors: Hiep X. Nguyen Mai Phuong Ho

Vaccination ranks among the most consequential interventions in medicine, yet cold-chain dependence, sharps hazards, needle phobia, and reliance on trained vaccinators limit coverage where vaccine-preventable mortality is highest. Microneedle patches deposit antigen into the antigen-presenting-cell-rich epidermis and upper dermis through projections that penetrate the stratum corneum without reaching dermal nociceptors. Such targeting yields immunogenicity matching or exceeding intramuscular injection at a fraction of the antigen mass, with dose-sparing up to six-fold recorded for influenza, polio, and SARS-CoV-2 antigens. Solid-state formulation converts that immunological advantage into a logistical one, since polymeric matrices preserve potency for as long as two years at ambient temperature, removing the refrigeration infrastructure that consumes significant delivery cost. Six microneedle types have reached preclinical or clinical maturity, with dissolving microneedle patches the most advanced. Two trials provide the clinical evidence: a Phase I influenza study showing non-inferior antibody responses and successful self-application, and a Phase I/II measles–rubella trial in The Gambia reaching 93% measles and 100% rubella seroconversion in infants without related serious adverse events. Engineering advances currently include an automated printer producing thermostable mRNA–lipid nanoparticle patches that retain bioactivity for six months at ambient temperature, the first intradermal self-amplifying RNA patch, and quantum-dot on-body immunization records. Sterility assurance, dose uniformity, nucleic acid integrity within solid matrices, and fragmented regulatory guidance remain the challenging obstacles, alongside a clinical pipeline concentrated on few antigens. This review integrates the mechanistic, materials, manufacturing, clinical, regulatory, and global health dimensions of the field to guide translation and equitable deployment.

Pharmaceutics doi: 10.3390/pharmaceutics18081021

Authors: Denis E. Yakobson Mikhail N. Zharkov Oleg A. Kulikov Vasilisa I. Kulikova Vladislav S. Bobrov Aleksey O. Makarov Ekaterina P. Brodovskaya Larisa A. Balykova Ran Yan Nikolay A. Pyataev

Background/Objectives: Combining chemotherapy with local magnetic hyperthermia (MHT) is promising because heating tumor tissue can increase cell damage, sensitize cells to cytostatic drugs, impair DNA repair, and change tumor microenvironment permeability. This creates conditions for enhancing the antitumor efficacy of chemotherapy while potentially reducing systemic toxicity. The aim of this study was to evaluate the efficacy of MHT with Zn0.2Mn0.8Fe2O4@OA nanoparticles alone and in combination with cisplatin in a Lewis lung carcinoma model. Methods: Four types of magnetic nanoparticles were synthesized and characterized: Fe3O4 and Zn0.2Mn0.8Fe2O4 coated with oleic acid (OA) or SiO2-NH2. Their physicochemical and magnetothermal properties, cytotoxicity, reactive oxygen species generation, and biodegradation in vivo were evaluated. Antitumor efficacy was studied in C57Bl/6 mice with LLC tumors after intratumoral administration of nanoparticles and two MHT sessions (100 kHz, 8 kA/m, 30 min). In combination therapy, ZnMn@OA and cisplatin at doses of 9 or 18 mg/kg were used. Results/Conclusions: Zn0.2Mn0.8Fe2O4@OA combined efficient heating, biodegradation, and the most pronounced effect among the MHT-alone groups, although MHT without chemotherapy did not provide sustained inhibition of tumor growth or a significant increase in survival. The combination of Zn0.2Mn0.8Fe2O4@OA-MHT with cisplatin 9 mg/kg produced the best therapeutic outcome: median survival increased significantly by two fold compared with the control group and by 1.8-fold compared with the chemotherapy-alone group at the comparable cisplatin dose. This regimen also stabilized body weight, reduced systemic toxicity, and restored RBC, HGB, and HCT parameters to the level of healthy animals by day 7 of the experiment. These data confirm the potential of MHT as a chemosensitizing approach that improves the efficacy and tolerability of cisplatin therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18081020

Authors: Amedeo De Nicolò Alessandra Manca Edoardo Ceraolo Giulio Luca Rosboch Antonio Toscano Luca Neitzert Eleonora Balzani Jessica Cusato Alice Palermiti Giorgia Giuseppina Montrucchio Antonio D’Avolio

Background/Objectives: Local anesthetics (LAs) are used in a variety of different contexts, from loco-regional anesthesia to analgesia. In cardiothoracic surgery, fascial plane blocks are deserving of attention, including erector spinae plane block (ESPB). In this context, ropivacaine is convenient, due to its peculiar pharmacokinetic/pharmacodynamic properties. Nevertheless, LAs can still cause systemic toxicity (LAST), due to erroneous injection and/or variable systemic adsorption/distribution. This interindividual pharmacokinetic variability can be intensified by the tendency to use a fixed ropivacaine dose in ESPB. The aims of this work were investigating systemic exposure to ropivacaine during ESPB in the context of cardiac and thoracic surgery, comparing it to the literature-reported maximum tolerated concentrations, and identifying potential predictors of exposure. Methods: Patients receiving ultrasound-guided injection of 40 mL of ropivacaine 0.375% solution for ESPB were enrolled. Arterial blood was sampled at 5, 15, 30, 45, 60, 120, and 180 min after the injection, and total and free concentrations of ropivacaine were determined by means of LC-MS/MS analysis of arterial plasma. Results: Concentrations showed wide variability, particularly during the first hour post-dose. Significant differences were observed between patients undergoing cardiac and thoracic surgery, with the latter showing higher concentrations, potentially above the cutoff values predictive of LAST. Pharmacokinetic differences were mainly explained by anthropometric (body weight and BSA) and clinical/hemodynamic (NYHA score) characteristics. Conclusions: This study shows that about 5% of patients could reach arterial plasma concentrations of ropivacaine above the cutoff level for LAST after ESPB with a 150 mg dose. Considering patients’ weight could be beneficial to maintain exposure below the toxicity cutoff.

Pharmaceutics doi: 10.3390/pharmaceutics18081019

Authors: Sarfaraz K. Niazi

Background/Objectives: Nanoparticle carriers can enhance cancer immunotherapy by improving tumor delivery, activating innate immune responses, and remodeling tumor microenvironments. This review evaluates four categories of formulations: poly(lactic-co-glycolic acid) (PLGA) nanoparticles, mesoporous silica nanoparticles (MSNs), magnetic or iron oxide nanoparticles (MNPs), and covalent organic frameworks (COFs). Methods: A reproducible PubMed audit identified 568 records. A rule-assisted title-and-abstract screen, followed by verification, removed 320 reviews, non-primary publications, and reports lacking qualifying in vivo formulation evidence. Of 248 potentially relevant reports, 15 primary studies were selected as representative examples; the remaining 233 were not classified as ineligible but were not selected as representative examples. These reports yielded 16 formulation-level records. One author conducted screening and extraction without protocol registration, duplicate review, or formal risk-of-bias scoring. Results: PLGA demonstrates the most robust polymer-level regulatory and manufacturing precedent; however, it remains limited by cargo instability, burst release, and challenges associated with process transfer. Biodegradable mesoporous silica nanoparticles (MSNs) facilitate pore-based protection and cytosolic delivery of cyclic dinucleotides, although their degradation and clearance are dependent on formulation specifics. Magnetic nanoparticles (MNPs) integrate magnetic targeting, imaging, and hyperthermia capabilities but necessitate formulation-specific magnetic characterization, field dosimetry, and repeated-dose safety assessments. Covalent organic frameworks (COFs) provide extensive stimulus-responsive and catalytic functionalities but exhibit the least mature evidence concerning biodegradation, scalable manufacturing, and independent reproducibility. Efficacy data across different studies were not pooled due to heterogeneity in models, schedules, comparators, and tumor-growth-inhibition formulas. Conclusions: The evidence does not endorse a universal platform ranking. Translation depends on standardized immune endpoints, explicit efficacy formulas, quantitative biodistribution assessments, mechanism-confirming experiments, repeat-dose toxicology studies, scalable manufacturing processes, and independent replication. The resulting evidence map serves as a descriptive and hypothesis-generating tool rather than a meta-analysis or clinical-priority scoring system.

Pharmaceutics doi: 10.3390/pharmaceutics18081018

Authors: Saurabh Tiwari Katarzyna Mądra-Gackowska Marcin Gackowski Łukasz Szeleszczuk

Background/Objectives: Predicting blood–brain barrier (BBB) permeability remains a major challenge in central nervous system (CNS) drug discovery. Three-dimensional (3D) conformer-derived molecular descriptors are often proposed as improvements over conventional two-dimensional (2D) topological representations; however, their incremental predictive value remains unclear. Methods: Here, we systematically evaluated six molecular feature sets comprising curated 2D Mordred descriptors, Morgan/ECFP4 fingerprints, 3D conformer-derived descriptors, and their combinations, using LightGBM regression on the B3DB benchmark dataset (1054 compounds with experimental logBB values). The model performance was assessed using 30 independent random splits and 30 Murcko scaffold-based splits. Results: The 2D descriptor model achieved mean test R2 values of 0.567 ± 0.060 and 0.418 ± 0.085 under random and scaffold splitting, respectively, whereas the 3D descriptor model performed substantially worse (0.430 ± 0.066 and 0.298 ± 0.097, respectively). Incorporating 3D descriptors into the 2D feature set yielded only a marginal improvement under random splitting (+0.008 R2; p = 0.039), which disappeared during scaffold-based validation (p = 0.599). Similarly, adding 3D descriptors to the combined 2D + fingerprint representation did not yield any significant benefits. Fingerprints alone exhibited pronounced scaffold fragility, with the mean R2 decreasing from 0.441 to 0.312 between the random and scaffold evaluations. SHAP analysis identified TopoPSA (NO) as the dominant predictor (mean |SHAP| = 0.332), highlighting the central role of desolvation in BBB permeation. Applicability domain analysis showed that 94.8% of the test compounds fell within the structural coverage of the training set. Conclusions: Overall, on the B3DB benchmark and under the descriptor implementation evaluated here, 3D conformer descriptors provided limited incremental value and no scaffold-robust advantage over well-curated 2D molecular representations for BBB permeability predictions.

Pharmaceutics doi: 10.3390/pharmaceutics18081017

Authors: Simona Hisirová Patrícia Hudecová Vanda Hajdučková Stanislav Lauko Nikola Dančová Lívia Mačák Oksana Velgosova Peter Paľove-Balang Ján Király

Background: This study evaluated the antibacterial, antibiofilm, and biofilm eradication activities of biogenically synthesized silver nanoparticles (AgNPs-L, AgNPs-R) mediated by extracts of Lavandula angustifolia and Salvia rosmarinus, individually and in combination with their respective essential oils (EOs) and ampicillin (AMP), against a multidrug-resistant biofilm-forming Staphylococcus equorum strain. Physicochemical characterization confirmed the successful biosynthesis of spherical AgNPs-L (10–25 nm) and AgNPs-R (5–15 nm). Individual treatments exhibited distinct antibacterial activity, with MIC values of 25 µg/mL for AgNPs-L, 12.5 µg/mL for AgNPs-R, and 0.1% (v/v) for both EOs; however, they showed no ability to eradicate preformed biofilms. Dual AgNPs/EO combinations at subinhibitory concentrations demonstrated borderline additive effects (FICI = 0.501) and significantly potentiated antibacterial and antibiofilm activity compared with individual treatments. Triple AgNPs/EO/AMP combinations exhibited the most pronounced biological effects, with predominantly additive interactions depending on AMP concentration. Lavender-based triple combinations achieved up to 63.9% inhibition of planktonic growth, 78.1% prevention of biofilm formation, and 37.3% eradication of mature biofilms, whereas rosemary-based combinations resulted in 57.1%, 69.4%, and 42.7% inhibition, respectively. These findings highlight the potential of multi-component systems integrating biogenic nanomaterials, phytochemicals, and conventional antibiotics as a promising strategy to enhance antimicrobial efficacy against persistent biofilm-forming non-aureus staphylococci.

Pharmaceutics doi: 10.3390/pharmaceutics18081016

Authors: Cláudia S. Paiva Hadi El Radi Diogo Chagas Kévin Grollier Johan Cailletaud Sébastien Delacroix Paolo Gabaldi Tiago Q. Faria Cristina Peixoto

Background: Process analytical technology (PAT) enhances product quality by monitoring and controlling critical quality attributes (CQAs), offering better insights into process performance. Raman spectroscopy is a promising PAT tool to support the development of control systems for continuous and automated processes. In this study, Raman spectroscopy was used to monitor in real-time ultra- and diafiltration (UF/DF) of adeno-associated virus (AAV) and lentiviral vector (LV). Method: The incorporation of a Raman probe in a tangential flow filtration (TFF) system, when operated in open loop, enabled spectral acquisition during a five-fold concentration of clarified bulk followed by a buffer exchange to PBS with five diafiltration volumes. Results: Principal Component Analysis reduced the dimensionality of the training set, and a Partial Least Squares model was developed for each parameter in each phase. After validation, Raman monitoring platforms showed good performance in predicting CQA of both viral vectors. Batch-to-batch variability was identified as the principal source of spectral variation, and its impact on UF/DF operation was observed. Discussion: This study emphasises the potential of Raman spectroscopy as a PAT for monitoring and control strategies in the downstream processing of viral vectors. Conclusions: These monitoring platforms enhance understanding of TFF’s operation and enable timely decision making by providing real-time information on multiple parameters.

Pharmaceutics doi: 10.3390/pharmaceutics18081015

Authors: Hala Rayya Raghad Alsheikh Dániel Nemes Lajos Nagy Géza Regdon Ildikó Bácskay Krisztián Pamlényi Katalin Kristó

Background/Objectives: The buccal mucosa offers a promising non-invasive route for systemic drug delivery, particularly for hydrophilic compounds like captopril (CAP), which exhibit low permeability and are subject to gastrointestinal instability and first-pass metabolism. This study aimed to develop and characterize captopril-loaded, chitosan-based mucoadhesive buccal films with different permeation enhancers and to evaluate their physicochemical properties, drug release, cytocompatibility, and in vitro transport across a TR146 buccal epithelial cell model. Methods: Films were prepared by the solvent-casting method using chitosan as the film-forming polymer. Different enhancers were investigated, including organic acid salts of chitosan (ascorbate, citrate, and lactate) and chemical permeation enhancers (sodium lauryl sulfate, polyethylene glycol 400, Span 20, and EDTA). Results: The resulting films exhibited acceptable thickness, moisture content, appropriate mechanical properties, and good mucoadhesive strength. In vitro dissolution studies demonstrated rapid CAP release, with >50% released within 15 min and near-complete release by 180 min across all formulations. Cytotoxicity assessment via a Neutral Red uptake assay in TR146 cells confirmed high cell viability (>81%) after 4 h of exposure, indicating good biocompatibility. In vitro permeation experiments revealed that films prepared with chitosan ascorbate and chitosan lactate enhanced CAP transport compared to other formulations, achieving the highest flux and apparent permeability coefficients. Conclusions: These findings demonstrate that chitosan ascorbate and lactate salts effectively improve the buccal permeability of captopril while maintaining good film properties and biocompatibility. This work highlights the potential of chitosan ascorbate- and lactate-based mucoadhesive films as an efficient platform for the buccal delivery of CAP.

Pharmaceutics doi: 10.3390/pharmaceutics18081014

Authors: Xinxin Deng Dandan Mu Fei Song Yeqing Miao Qiang Yin Hailong Yin

Background/Objectives: Oral solid dosage forms of traditional Chinese and ethnic medicines are currently undergoing modernisation. The objective of this study is to explore the scope for formulation variation arising from batch-to-batch fluctuations in intermediates, and to identify the factors influencing key quality attributes of honey-containing tablets. In this regard, a machine-learning-based predictive model is being formulated that will integrate and analyse formulation factors and the results characterised by the SeDeM expert system. Utilising the SeDeM index as a mediating variable, the study endeavours to establish a comprehensible and predictable stepwise research pathway to provide a foundation for industrial-scale upscaling. Methods: Twelve SeDeM expert systems were utilised to characterise honey-containing granules for formulation screening, to evaluate their suitability for use in traditional Chinese medicine honey-containing tablet systems, and to identify key limiting factors and the feasibility space affecting the quality of the final product; Based on the QBD philosophy, a TriAD (Tri-criterion Adaptive Design) design scheme was proposed, integrating the horizontal balance of orthogonal designs, the spatial coverage of uniform designs, and the parameter estimation efficiency of D-optimal designs into the experimental layout of the formulation feasibility space; Through further data aggregation, a multi-layer feature set comprising four formulation factors, six SeDeM indicators, and three critical quality attributes (CQAs) was constructed. The mediating effects of the SeDeM indicators were revealed through different pathways involving 37 combinations of simple, linear, and Bootstrap models. Furthermore, 180 linear and non-linear machine learning models (comprising 12 categories of algorithms) were trained to predict formulation and CQA outcomes, ultimately completing the design space mapping and validation. Results: The results show that the SeDeM parameters effectively bridge the CQA results of different honey formulations, with these indicators acting as selective mediators between formulation factors and CQAs. Compared with a pure data model relying solely on raw formulation variables, the introduction of SeDeM knowledge, combined with high-information-content samples obtained via TriAD, improved the predictive performance and robustness of the SeDeM–ML hybrid model in terms of disintegration time and hardness; its R2_LOO increased by 0.267 and 0.510, respectively, and the overall predictive space was significantly expanded. Experimental validation was conducted using formulations within the design space predicted by the optimal model; the results showed that both the prediction bias and the relative standard deviation were less than 5 percent. Conclusions: The present study demonstrates that SeDeM can not only be used to evaluate formulations of honey-containing TCM tablets but also serves as an intermediary bridge linking formulation factors, granule-mechanism variables, and tablet quality outcomes. TriAD, in turn, further translates the QbD philosophy into an actionable formulation space design, thereby providing a development pathway for honey-containing tablets that combines interpretability, predictability, and QbD consistency, and offers new insights for the industrial application of oral TCM preparations.

Pharmaceutics doi: 10.3390/pharmaceutics18081013

Authors: K. P. D. H. Pathirana D. A. Upeka Chathurangani Julian Vlad D. M. W. S. Dissanayake Yasiru Vindula Alwis G. Mashooda Jayah K. P. S. S. Pathirana Dinusha Nishani Udukala Nishal M. Egodawaththa Jason P. Farrah Nasri Nesnas Medha Jaimini Gunaratna

Background/Objectives: Diabetes mellitus is a common endocrine disorder characterized by hyperglycemia, which is increasing steadily all over the world. Current medications are limited due to lower efficacy and side effects; therefore, the introduction of novel compounds is crucial to improve therapeutic outcomes for diabetic patients. The objective of this study was to synthesize and evaluate 2-phenyl-3H-quinazolin-4-one and its derivatives for the glucose uptake across the yeast cell membranes, alpha-amylase inhibition, and alpha-glucosidase inhibition in silico and in vitro. Methods: The synthesis of 2-phenyl-3H-quinazolin-4-one and its derivatives was carried out by oxidative cyclocondensation of 2-aminobenzamide (anthranilamide) with various benzaldehydes in the presence of aqueous iron (III) chloride. The synthesized compounds were characterized by melting point determination and spectroscopic techniques, including FTIR, NMR and HRMS. Results: In the glucose uptake by yeast assay, compound 3f had a lower 50% glucose uptake value of 24.02 ± 0.96 mM, compared with the standard drug metformin (25.99 ± 2.41 mM). The alpha-amylase inhibition bioassay showed that compound 3e exhibited very potent inhibition, with an IC50 of 0.43 ± 0.05 mM, compared with the standard drug acarbose (0.53 ± 0.21 mM). Compound 3l exhibited strong alpha-glucosidase inhibition, with an IC50 of 0.42 ± 0.10 mM, compared with the standard drug acarbose (0.29 ± 0.50 mM). Coclusions: These findings suggest the potential of synthesized 2-phenyl-3H-quinazolin-4-one derivatives, 3a–o, as promising candidates for managing diabetes mellitus. However, further studies are required to validate their efficacy and determine their mechanisms of action.