Molecular Pharmaceutics最新文献

The rapid development of mRNA vaccines during the COVID-19 pandemic has highlighted the critical role of lipid nanoparticles (LNPs) as delivery systems. The advantages of prefilled syringes (PFSs) in mRNA-LNP administration are widely recognized. The compatibility of mRNA-LNP drugs with tungsten in PFSs has not yet been investigated. In this study, we used polyadenylic acid (Poly A) as an mRNA model to conduct accelerated stability experiments under conditions of 4 °C, 25 °C, and light exposure, examining the effects of three tungsten sources (commercial salts or tungsten extracted from syringe pins) on the physicochemical properties of LNPs. Additionally, enhanced green fluorescent protein (eGFP)-mRNA and Poly A were compared to validate the changes in bioactivity. Our findings revealed that tungsten significantly increased the particle size and polydispersity index (PDI) of Poly A-LNPs, while reducing zeta potential and encapsulation efficiency. Transmission electron microscopy (TEM) further demonstrated that tungsten-induced structural damage to Poly A-LNPs. eGFP-LNPs spiked with 50 ppm tungsten extract completely lost activity after 6 weeks of storage at 25 °C, even though they exhibited greater physicochemical stability than Poly A-LNPs. Light exposure, while having no significant impact on physicochemical parameters, substantially diminished LNP bioactivity. Subsequent nucleic acid integrity testing of tungsten-spiked eGFP-mRNA revealed that tungsten caused minimal changes in physicochemical properties, such as particle size and PDI, under real-world conditions, but significantly compromised eGFP-mRNA integrity. This suggests that mRNA integrity, rather than physicochemical metrics such as particle size, PDI, or encapsulation efficiency, is the critical quality attribute determining mRNA-LNP bioactivity.

CXCR4 is overexpressed in various malignancies and represents an attractive target for PET imaging. However, currently available peptide-based tracers often exhibit rapid clearance and a narrow imaging window, which limit their clinical implication. Here, we report the design and preclinical characterization of a ⁶⁸Ga-labeled dimeric cyclic peptide, implementing a large-volume linker strategy aimed at achieving an optimized balance between receptor affinity and tumor retention for CXCR4-targeted PET imaging. The tracer was synthesized, radiolabeled with the ⁶⁸Ga³⁺ ion, and evaluated for radiochemical purity. In vitro stability, binding affinity, CXCR4-specific cellular uptake, pharmacokinetics, biodistribution, and PET/CT imaging were also assessed. The tracer showed high radiochemical purity and excellent stability. Although its binding affinity was moderate (IC₅₀ = 161.5 nM), the tracer exhibited clear CXCR4-specific uptake and sustained tumor retention, with PET-derived tumor uptake of 3.4-3.8%ID/g between 30 and 240 min and tumor-to-muscle ratios increasing from ∼10 to ∼62 over the same period. However, notable hepatic uptake was observed, which may be attributed to the peptide size and moderate hydrophobicity. Further structural optimization such as PEGylation or scaffold minimization may reduce hepatic uptake and enhance the clinical applicability.

Most studies on fibroblast activated protein (FAP)-targeted radiopharmaceuticals focus on administered radioactivity, often overlooking the impact of a molar dose on tumor-targeting and off-target accumulation. Here, we investigate the effect of molar dose on biodistribution and pharmacokinetics using [⁶⁸Ga]Ga-FAPI-04 PET and systematically evaluated two FAP-targeted dimers, DOTAGA.(SA.FAPi)₂ and DOTAGA.Glu.(FAPi)₂, in a 4T1 syngeneic tumor model. Dynamic PET imaging confirmed a clear molar dose-dependent effect on tumor uptake, tumor-to-organ ratios, and organ pharmacokinetics with lower molar doses prolonging tumor retention. Comparative analyses across multiple molar doses revealed that DOTAGA.Glu.(FAPi)₂ achieved comparable tumor uptake to DOTAGA.(SA.FAPi)₂ but exhibited significantly reduced liver accumulation. An optimal molar dose range of 8–32 nmol/kg was identified, balancing maximal tumor uptake with reduced off-target exposure. At this optimized dose, [¹⁷⁷Lu]Lu-DOTAGA.Glu.(FAPi)₂ demonstrated therapeutic efficacy in 4T1 tumor-bearing mice with limited systemic toxicity. These results establish molar dose optimization as a broadly applicable strategy for accurately evaluating and comparing FAP-targeted radiopharmaceuticals and provide a methodological framework to guide future preclinical development and translational studies.

mRNA-based therapeutics represent a major advancement in modern medicine, offering programmable and nonintegrating treatment options for infectious diseases, malignancies, and hereditary disorders. This review addresses the chronological evolution, structural optimization, and delivery challenges of mRNA drugs, highlighting developments such as nucleoside modifications and lipid nanoparticle (LNP) platforms that improve the stability and promote cellular entry. Comparative analysis highlights the benefits of mRNA over DNA-, siRNA-, and protein-based medicine in safety, scalability, and rapid rearrangement. Applications vary from COVID-19 vaccines to individualized cancer immunotherapy and protein replacement strategies. New methods, including self-amplifying mRNA (saRNA), CRISPR-Cas9 gene editing, and tissue-specific delivery systems, enhance the therapeutic potential. While mRNA technology faces challenges in terms of immunogenicity, multiple dosing, and durability of safety considerations, it offers unparalleled precision, transient expression, and swift manufacturability. This review emphasizes the comparative design principles of mRNA delivery systems, bridging formulation innovation with translational biomedical applications. By integrating lipid-based and nonlipid nanocarrier insights, it highlights critical advances shaping next-generation mRNA therapeutics.

Radiolabeled fibroblast activation protein inhibitors (FAPIs) have limited clinical translation potential due to suboptimal tumor retention. In this study, we engineered novel polymeric FAPI tracers to enhance molecular affinity, tumor uptake, and retention. These tracers were comprehensively evaluated for their binding specificity, biodistribution characteristics, and potential applications in both preclinical PET/CT imaging and preliminary clinical studies involving human volunteers, which revealed that the ⁶⁸Ga radiolabeling of polymerized FAPIs enhanced all three parameters. For therapeutic evaluation, we compared the tumor growth inhibition effects of [¹⁷⁷Lu]Lu-FAPI-A1 and [¹⁷⁷Lu]Lu-FAPI-46. FAPI-A1 exhibited a high FAP affinity and specificity. In HT-1080-FAP tumor models, [⁶⁸Ga]Ga-FAPI-A1 demonstrated significantly higher tumor uptake, longer retention, and slower clearance compared to [⁶⁸Ga]Ga-FAPI-46. [¹⁷⁷Lu]Lu-FAPI-A1 induced a significant tumor growth suppression. Clinical uptake of [⁶⁸Ga]Ga-FAPI-A1 mirrored that of FAPI-46. FAPI-A1 exhibits enhanced binding affinity, which improves tumor uptake and prolongs the retention of [¹⁷⁷Lu]Lu-FAPI-A1, supporting its theranostic potential.

Adjuvants, as vital components of vaccines, possess the capacity to augment the intensity, breadth, and persistence of immune responses. Traditional alum adjuvants are incapable of inducing Th1 cellular immunity. To enhance the efficacy and alter the type of immune responses triggered by alum adjuvants, we constructed a Pickering emulsion platform stabilized by Chinese yam polysaccharide-loaded aluminum hydroxide nanoparticles (C-AlPE). The Chinese yam polysaccharide (CYP) was utilized as the immunopotentiator, and the aluminum hydroxide nanoparticles (Al NPs) loaded with CYP were employed as solid nanoparticle stabilizers. The C-AlPE as an adjuvant for the H9N2 vaccine elicited a robust humoral immune response. Compared to commercial Algel adjuvant and ISA206 adjuvant, the C-AlPE adjuvant obviously promoted the generation of CD4⁺ and CD8⁺ T cells and increased the production of Th1- and Th2-type cytokines, thereby stimulating a balanced Th1/Th2 immune response. In addition, the C-AlPE adjuvant exhibited great biosafety in vivo against the H9N2 vaccine. RNA-seq analysis of the spleen indicated that the C-AlPE adjuvant modulated complex signaling pathways, thereby activating innate immune responses and subsequently adaptive immune responses. Our findings highlighted the considerable potential of C-AlPE as a safe and effective vaccine adjuvant, providing valuable insights for the development of novel polysaccharide- and alum-based adjuvants in a Pickering emulsion formulation, with potential applications in veterinary vaccines.

Acute liver injury (ALI), often triggered by an acetaminophen (APAP) overdose, is characterized by severe oxidative stress, inflammation, and hepatocyte apoptosis. Current therapies, such as N-acetylcysteine (NAC), are limited by narrow treatment windows, highlighting the need for more effective antioxidant strategies. In this study, cerium oxide nanoparticles (CeO₂ NPs, nanoceria) were synthesized and comprehensively characterized using the transmission electron microscopy (TEM), dynamic light scattering method (DLS), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) to confirm their branched morphology, high crystallinity, and mixed Ce³⁺/Ce⁴⁺ valence states. Their enzyme-mimetic antioxidant activities were evaluated through superoxide, hydrogen peroxide, and hydroxyl radical scavenging assays. Nanoceria exhibited excellent cytocompatibility and effectively suppressed the generation of lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) and lipid peroxidation and caspase-3-mediated apoptosis in macrophages. They also downregulated pro-inflammatory mediators (nitric oxide synthase (iNOS), TNF-α, IL-1β, and NLRP3) while enhancing anti-inflammatory markers (Arg1 and IL-10). In an APAP-induced ALI mouse model, nanoceria preferentially accumulated in the liver, alleviated oxidative stress and inflammation, and significantly reduced aspartate aminotransferase (AST) levels, showing hepatoprotective efficacy comparable to NAC. Nanoceria protect against APAP-induced ALI via synergistic antioxidative and anti-inflammatory mechanisms based on reversible Ce³⁺/Ce⁴⁺ redox cycling. These findings underscore nanoceria’s potential as a next-generation nanotherapeutic for oxidative stress-related liver diseases.

Proteolysis-targeting chimeras (PROTACs) are transforming targeted therapeutics by enabling the selective and catalytic degradation of disease-associated proteins, offering new treatment avenues in oncology, neurodegeneration, immunology, and infectious diseases. However, their clinical translation is limited by poor membrane permeability, low oral bioavailability, suboptimal pharmacokinetics, a narrow range of useable E3 ligases, emerging resistance mechanisms, and safety concerns associated with their use. Nanocarrier-based delivery systems, including liposomes, polymeric nanoparticles, dendrimers, exosomes, and metal–organic frameworks, have emerged as promising solutions to these challenges. By enhancing solubility, protecting PROTACs from enzymatic degradation, improving cellular uptake, and enabling controlled stimuli-responsive release, nanocarriers can significantly improve pharmacological performance and tumor accumulation and reduce off-target toxicity. This review critically explores the intersection of PROTAC technology and nanocarrier-enabled delivery, summarizes key physicochemical and biological barriers, highlights recent advances in nanocarrier design and preclinical applications, and discusses the integration of artificial intelligence in rational PROTAC development. The persistent challenges of endosomal escape, tumor heterogeneity, immunogenicity, and regulatory hurdles were addressed, and future directions were proposed for optimizing nanoenabled PROTAC therapies to fully realize their clinical potential.

Tuberculosis (TB) remains a major global health challenge, aggravated by limitations of current chemotherapeutic regimens, including poor oral bioavailability, systemic side effects, and patient noncompliance. This study aimed to develop and evaluate macrophage-targeted, mannose-conjugated solid lipid nanoparticles (Mn-ETB-SNs) for enhanced oral delivery of ethylbutol (ETB), a first-line anti-TB drug. The Mn-ETB SNs were fabricated using high-pressure homogenization followed by surface mannosylation through Schiff’s base formation between mannose and amine-functionalized nanoparticles. The prepared formulations were characterized for particle size, ζ-potential, drug loading, entrapment efficiency, morphology, and stability. In vitro release, GI stability, cytotoxicity, and cellular uptake studies using J774A.1 macrophages were conducted. Further, in vivo pharmacokinetic and biodistribution studies were performed in Sprague–Dawley rats. The optimized Mn-ETB-SNs exhibited a uniform particle size of approximately 491 nm, a high entrapment efficiency of around 84%, and spherical morphology with stable physicochemical properties under varied storage and GI conditions. Mannosylation significantly enhanced macrophage uptake by 2.02-fold compared to unconjugated nanoparticles, as confirmed through fluorescence-activated cell sorting (FACS) and fluorescence microscopy. In vivo pharmacokinetic studies demonstrated an 8.5-fold increase in ETB bioavailability with Mn-ETB-SNs compared to the pure drug, accompanied by prolonged circulation and reduced hepatic metabolism. Biodistribution analysis revealed preferential and sustained lung accumulation, with Mn-ETB-SNs achieving 4.74-fold higher pulmonary concentrations at 48 h compared to free drug, owing to mannose receptor-mediated uptake by alveolar macrophages. Collectively, the findings highlight the potential of orally administered Mn-ETB-SNs as a promising nanocarrier system for targeted TB therapy. The developed formulation offers improved bioavailability, site-specific drug delivery, and enhanced pulmonary targeting, addressing key limitations of conventional TB treatment.

Understanding differences in compression behavior between amorphous and crystalline forms of organic drugs provides valuable insights into how amorphization affects tabletability, which is critical for the rational development of pharmaceutical tablets. In this work, we evaluate the compaction properties of both solid forms across several structurally diverse model drugs and found that the tabletability profiles of amorphous forms cluster within a narrower range than those of their crystalline counterparts. Moreover, the differences observed at high compaction pressures are primarily governed by variations in interparticulate bonding strength, as supported by infrared spectroscopy. Finally, we discuss the broader implications of these findings for the design and development of amorphous solid dispersions in tablet formulations.