Molecular Pharmaceutics最新文献

In preclinical settings, flux experiments can serve multiple purposes, such as (1) to select a formulation that provides higher flux, (2) to establish use-times for supersaturating formulations such as spray dried dispersion suspensions, and (3) to explain variation in exposures in in vivo experiments. In the current manuscript, a total of five internal model compounds are shown to portray the scenarios where flux measurements have been used. The overarching aim of this study was to provide a decision-making framework for identifying optimal formulations based on flux. Two different diffusion set ups were explored: 96-well PermeaPad plates and the Pion μFLUX diffusion system. The 96-well PermeaPad plates were initially tested as a drug-sparing method for conducting the flux assay. However, undetectable concentrations in the receiver compartment hindered its application across compounds, needing to pivot to a larger size diffusion set up, using the Pion μFLUX and measuring concentrations by mass spectrometry. This study highlights the advantages and limitations of each diffusion setup. It underscores the importance of amorphous solubility values and MDCK passive permeability in determining whether PermeaPad plates will produce detectable concentrations in the receiver compartment through mass spectrometry. This study displays a good correlation between in vitro flux experiments and in vivo exposures in animal species and shows drawbacks of using dissolution data alone. Lastly, this work emphasizes the importance and potential use of flux assays to screen formulations or to explain formulation differences seen in vivo.

Tumor hypoxia promotes angiogenesis, dysfunctional vascular formation, and the epithelial-to-mesenchymal transition phenotype. We propose that a CXCR4 inhibitor (AMD3100) could decrease cancer stemness and improve the efficacy of the anticancer drug gemcitabine (GEM) in targeting hypoxic pancreatic ductal adenocarcinoma (PDAC) cells. In this study, we synthesized a hypoxia-responsive polymer containing poly(lactic acid)-diazobenzene-poly(ethylene glycol) to produce hypoxia-responsive polymersomes. Cell viability experiments showed that AMD3100 enhanced the cytotoxicity of GEM-encapsulated polymersomes under hypoxic conditions compared to normoxia. The combined treatment significantly elevated pro-apoptotic BAX mRNA levels and reduced antiapoptotic BCL2 mRNA levels. Additionally, the combination therapy decreased the size of cancer cell spheroids from PANC1 and patient-derived cells.

The clinical translation of tranexamic acid (TA) for melasma is limited by its hydrophilicity and crystallization tendency, which hinder epidermal drug accumulation in conventional formulations (<5% w/w). We developed a supersaturated emulsion-type gel (TAE Gel) containing 10% (w/w) TA using carbomer 940 (CP) as the matrix. Electrostatic interactions between TA and CP, including hydrogen bonding and ionic pairing, formed a kinetically stabilized TA@CP network that suppressed TA crystallization while enhancing transdermal delivery. The formulation maintained physical stability under stress conditions and showed no visible crystallization. Pharmacokinetic studies demonstrated superior epidermal bioavailability, with a peak dermal concentration 1.91-fold higher than commercial TA cream and minimal irritation. In UV/progesterone-induced melasma models, 10% TAE Gel significantly suppressed tyrosinase activity, restored antioxidant markers (SOD), and reduced lipid peroxidation (MDA), accompanied by normalization of hepatic biomarkers. Histological analysis confirmed melanin clearance and epidermal remodeling without cytotoxicity. Overall, this metastable supersaturation approach offers a promising strategy for high-payload transdermal delivery of hydrophilic actives. The TA@CP-based TAE Gel successfully addresses the formulation trade-off between dose escalation and physical stability, providing a safe and effective platform for topical melasma therapy and potentially other pigmentary disorders.

Macrophages play a crucial role in the pathophysiology of sepsis, serving as central regulators of both disease onset and progression. Among the therapeutic agents investigated to modulate macrophage-driven inflammation, compounds extracted from the roots of Panax ginseng have been distinguished for their potent anti-inflammatory properties. Recently, attention has shifted toward ginseng-derived exosome-like nanoparticles (GDEs) because of their ability to encapsulate diverse bioactive compounds with high stability and biocompatibility. In this study, we investigated the protective effects of GDEs against lipopolysaccharide (LPS)-induced septic shock, with a particular focus on macrophage-mediated mechanisms. GDEs isolated from Korean Panax ginseng exhibited a spherical morphology, high stability, and effective encapsulation of bioactive ginsenosides, including Rb1, Rg1, and Rg3. GDEs significantly attenuated LPS-induced inflammatory responses in macrophages by reducing toll-like receptor 4 (TLR4) glycosylation, thereby inhibiting LPS binding. This suppression of TLR4 glycosylation led to decreased production of nitric oxide and proinflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-α), as well as inhibition of intracellular reactive oxygen species accumulation and NF-κB activation. Furthermore, GDE treatment markedly improved survival and alleviated lung, liver, and spleen damage in an LPS-induced sepsis mouse model. In summary, these findings suggest that GDEs represent a promising nanomedicine strategy for sepsis prevention, offering targeted modulation of macrophage activity without apparent adverse effects.

Tumor vascular abnormalities can reduce drug and oxygen delivery and induce tumor resistance. However, it has recently been found that a few antiangiogenic drugs can achieve significant efficacy when combined with chemoradiotherapy, which is contrary to the effect of antiangiogenic drugs in clearing blood vessels. The theory of vascular normalization, originating from clinical observations, has explained this paradox and has become a promising strategy to overcome the bottleneck of tumor treatment, but there is a lack of innovative drugs. Based on the previous identification of the novel vascular normalization target EphrinB2, a synergistic strategy, “vascular normalization─chemotherapy” was proposed. Based on the principle of twin drugs in drug design and the controlled release strategy, we found a new type of vascular normalization-chemotherapy twin drug based on the active molecule QDAU5, which showed good membrane permeability, strong stability, rapid toxic payload release characteristics, and strong inhibitory activity against colorectal tumors. The study further enriches the theory of vascular normalization and promotes its expansion into the field of new drug development.

Bispecific immune checkpoint inhibitors (ICIs), with their ability to target multiple immune checkpoint molecules, demonstrate promising potential as carriers for radioimmunotherapy (RIT). We used ¹³¹I-labeled cadonilimab (AK104), a PD-1/CTLA-4 tetravalent bispecific antibody, as a novel radio-immunotherapeutic agent for the treatment of non-small-cell lung cancer (NSCLC) and explored its underlying therapeutic mechanism. SPECT/CT imaging was performed to evaluate the systemic and tumor distributions of [¹³¹I]I-AK104. The therapeutic effect of [¹³¹I]I-AK104 in tumor-bearing mice was evaluated by tumor growth monitoring and histopathology analysis. Imaging mass cytometry and single-cell RNA sequencing were used to analyze the changes in the tumor microenvironment after the [¹³¹I]I-AK104 treatment. [¹³¹I]I-AK104 exhibited good tumor-targeting retention ability and antitumor effect. It promoted CD8⁺ T cell infiltration and antigen-specific clonal expansion in the tumor microenvironment (TME). Furthermore, it induces immunogenic cell death (ICD) by recruiting neutrophils and triggering the release of reactive oxygen species (ROS) and nitric oxide (NO). Additionally, [¹³¹I]I-AK104 enhanced interactions among immune cell subsets, increasing the prevalence of the cell neighborhood (CN) composed of neutrophils, CD8⁺ T cells, and dendritic cells (DCs). The combination of ¹³¹I-based radiotherapy and bispecific ICIs remodels the TME and enhances the therapeutic efficacy in NSCLC. CD8⁺ T cell activation and neutrophil-mediated ROS and NO release are key mechanisms contributing to enhanced tumor cell killing.

T-cell dysfunction imposed by the tumor microenvironment limits the durability of cellular immunotherapies, motivating interventions that strengthen effector programs before infusion. Here we describe an ex vivo pretreatment that uses zinc nanoparticles (Zn NPs) as a transient intracellular Zn²⁺ modulator to condition T cells without altering receptor engineering. Brief priming with Zn NPs yielded T cells with consistently improved tumor-cell killing across effector-to-target ratios and increased IL-2, IFN-γ, and TNF-α secretion while avoiding rises in intracellular ROS. In murine melanoma models, adoptive transfer of primed T cells delayed tumor growth and prolonged survival without toxicity; combination with Galectin-9 blockade further enhanced control. Translational relevance was supported in an ovarian cancer patient-derived xenograft model, where primed CAR T cells achieved superior tumor control, showed greater intratumoral CAR T infiltration, and again benefited from Galectin-9 inhibition. Transcriptome profiling of primed T cells revealed coordinated remodeling of metal-ion response, consistent with reinforced signaling and stress resilience. This study establishes Zn NP–enabled priming as a practical, drop-in step for T-cell manufacturing that enhances effector function and cooperates with galectin-axis inhibition to overcome microenvironmental suppression and improve therapeutic potency.

Modulating the structure of nanoassemblies to achieve the desired properties remains a challenge. In this study, we present a strategy for regulating nanoassembly by introducing different numbers of polyethylene glycol (PEG) chains to control the self-assembly behavior of zinc phthalocyanine. The morphology, water solubility, and optical properties of nanoassemblies can be effectively tuned by adjusting assembly behaviors. Notably, zinc phthalocyanine nanoassemblies with four PEG chains (Pc4 NPs) showed the highest reactive oxygen species (ROS) generation efficiency and photothermal capacity. Furthermore, the incorporation of PEG chains enhanced the water solubility of nanoassemblies, thereby promoting their accumulation at the tumor site. This study provides a simple and feasible strategy for constructing nanomaterials with tunable optical properties.

Mesothelin (MSLN) is overexpressed in various solid tumors but limited in normal tissues, making it a promising candidate for molecular imaging and targeted therapy. Nanobodies, the smallest antibody derived fragments, are facile to modify for functional agent conjugation. With radionuclides, nanobody-based immunopositron emission tomography (immuno-PET) will provide valuable whole-body information for analysis of related target properties. The objective of this work was to evaluate the in vivo distribution peculiarity of ⁶⁸Ga-NOTA-MS3 as an MSLN-overexpressing tumor-directing probe. After rigorous quality control, xenograft models exhibiting differential MSLN expression were constructed for in vivo behavior evaluation of ⁶⁸Ga-NOTA-MS3. In addition, to improve the visualization of intra-abdominal tumors and mitigate potential radiation-related nephrotoxicity, a strategy involving preadministration of sodium maleate was employed to reduce the renal uptake of ⁶⁸Ga-NOTA-MS3. The nondecay corrected radiolabeling yield of ⁶⁸Ga-NOTA-MS3 was approximately 47%, and the radiochemical purity was >98%. Both immuno-PET imaging and biodistribution studies revealed that the uptake of ⁶⁸Ga-NOTA-MS3 was high in BxPC-3 and OVCAR-3 tumors, while HuH-7 tumors showed low uptake. These results aligned with the MSLN expression assessment. Moreover, 2 h postinjection of the imaging agent was identified as the optimal timing for immuno-PET. Furthermore, the preadministration of sodium maleate significantly reduced the renal accumulation of ⁶⁸Ga-NOTA-MS3 (from 61.14 ± 7.57%ID/g to 4.31 ± 1.31%ID/g), hence ameliorating the delineation of orthotopic pancreatic tumors. The study validates ⁶⁸Ga-NOTA-MS3 as an effective probe for the precise, noninvasive imaging of MSLN, which, in turn, allows for the accurate diagnosis of MSLN-expressing lesions.