Molecular Pharmaceutics最新文献

To combat multidrug resistance and cancer stem cell (CSC) persistence, we constructed a tumor-targeted nanoplatform integrating silver/copper alloy nanoparticles (Cu–Ag NPs) and camptothecin (CPT) nanocrystals for synergistic multimodal therapy. The nanocomposite was fabricated by stepwise assembly of CPT nanocrystals, a polydopamine coating, and functionalization with Cu–Ag NPs plus a tumor-mitochondria dual-targeting peptide. It exhibited a hydrodynamic diameter of ∼152.67 nm, high colloidal stability, favorable photothermal performance, and pH/NIR-responsive drug release. Under NIR irradiation, it showed potent and selective cytotoxicity against triple-negative breast cancer cells (IC₅₀ = 16.92 ± 0.22 μg/mL), with strong synergy (CI < 0.3) between inorganic Cu–Ag NPs and organic CPT. Actively targeting both cancer cells and mitochondria, it induced severe mitochondrial dysfunction─loss of MMP, ATP depletion, ROS burst, and mtDNA damage. Moreover, it acted as a potent cuproptosis inducer via exogenous copper, evidenced by FDX1 and DLAT downregulation (48.23% and 68.61%) and HSP70 upregulation (61.42%). Additional cell death pathways, including apoptosis, necrosis and pyroptosis, were also activated through nuclear DNA damage and plasma membrane rupture. Importantly, this nanoplatform effectively targeted stubborn breast CSCs, exhibiting an IC₅₀ as low as 13.70 ± 0.36 μg/mL─attributed to the mitochondrial targeting and subsequent inhibition of robust oxidative phosphorylation within CSCs, which rely more heavily on this pathway than on glycolysis compared to conventional cancer cells. In summary, this work presents a novel “multi-targeting” therapeutic strategy that orchestrates mitochondrial dysfunction, cuproptosis, apoptosis, and pyroptosis via a chemo-photothermal combination, offering a robust and broad-spectrum approach to eradicate both conventional resistant cancer cells and refractory CSCs.

The heterogeneous expression of tumor biomarkers limits the diagnostic performance of single-target imaging agents. Carbonic anhydrase IX (CAIX) is highly expressed in hypoxic regions of clear cell renal cell carcinoma (ccRCC) and multiple solid tumors, whereas prostate-specific membrane antigen (PSMA) is specifically upregulated in tumor-associated neovasculature. Both targets have been implicated in tumor metastasis and poor clinical outcomes. This study aimed to design and evaluate a novel bispecific PET tracer, [⁶⁸Ga]Ga-PCA, targeting both CAIX and PSMA, with the goal of achieving improved tumor-specific uptake. Subcutaneous xenograft models were established in nude mice by inoculation with OS-RC-2, PC3-PIP, and HEK-293 cells. PET/CT imaging and biodistribution studies were performed following intravenous administration of [⁶⁸Ga]Ga-PCA. Target specificity was evaluated via competitive blocking assays employing excess unlabeled ligand. Immunohistochemical staining was performed to validate the expression profiles of the targets within the tumors. After being labeled with gallium-68, [⁶⁸Ga]Ga-PCA showed favorable physicochemical properties, such as a high radiolabeling yield (>80%), radiochemical purity over 95%, good stability in vitro, and an albumin-binding rate of 93.44 ± 0.81%. PET/CT imaging revealed pronounced and specific tracer accumulation in both OS-RC-2 and PC3-PIP tumor models. In OS-RC-2 tumors (PSMA⁺/CAIX⁺), the SUVmax (13.10 ± 0.84) was higher than those of the single-target tracers [⁶⁸Ga]Ga-DOTA-NY104 (5.31 ± 0.77) and [⁶⁸Ga]Ga-PSMA (2.31 ± 0.49) at 60 min postinjection. An excess of unlabeled DOTA-NY104, a PSMA-targeted ligand, or a mix of the two ligands can block the uptake of [⁶⁸Ga]Ga-PCA. These results demonstrate that the tracer can bind to both targets at once. In conclusion, [⁶⁸Ga]Ga-PCA is a bispecific PET tracer that targets both hypoxic tumor cells and tumor neovasculature by binding to both CAIX and PSMA. The probe exhibited significant specificity, advantageous imaging contrast, and robust blocking validation, indicating its potential for molecular imaging of malignancies, including clear cell renal cell carcinoma (ccRCC).

The self-assembly of hyaluronic acid (HA) into stable nanoassemblies remains a significant challenge. To address this, we report a novel strategy utilizing l-histidine (His) as a molecular bridge to integrate HA with a zeolitic imidazolate framework-8 (ZIF-8). The key to this approach is the covalent conjugation of His to the HA backbone, which enables Zn²⁺ from ZIF-8 to coordinate with the imidazole groups of His. This coordination facilitates the ZIF-8-induced assembly of HA into stable hybrid nanoparticles (HA-His/ZIF-8 NPs). The resulting system synergistically combines the CD44-targeting capability of HA with the pH-responsive dissociation of ZIF-8. These NPs demonstrated a high doxorubicin (DOX) loading capacity (0.34 mg/mg) and encapsulation efficiency (76.8%). Importantly, they exhibited controlled drug release with significant pH-dependency, achieving a cumulative release of 50.2% under weakly acidic conditions (pH 5.0) compared to only 12.4% at physiological pH (7.4). In vitro studies confirmed the target-specificity of the DOX-loaded HA-His/ZIF-8 NPs, which were efficiently internalized by CD44-positive MKN-45 gastric cancer cells via receptor-mediated endocytosis, leading to a potent cytotoxic effect (IC₅₀ = 1.71 μg•mL^–1). In contrast, the efficacy was significantly lower in CD44-negative SNU-216 cells (IC₅₀ = 5.22 μg•mL^–1). This work highlights the strategic use of His as a bridge to create a synergistic HA-ZIF-8 platform, offering a powerful and promising approach to the targeted therapy of CD44-overexpressing cancers.

A solubilizing receiver medium has been documented to increase drug flux in vitro, but the mechanisms underlying this effect remain poorly understood. This study investigated these mechanisms and established a mathematical model to describe the increase in apparent permeability. Flow rate experiments were performed to quantify the individual boundary layer and membrane resistances associated with diffusion. The impact of nine solubilizing receiver additives, including surfactants, cyclodextrins, and bovine serum albumin, on the flux of griseofulvin was investigated. The increase in apparent permeability followed the rank-order, though not the magnitude, of the solubility enhancement in the receiver (Spearman’s ρ = 0.93, p < 0.001, n = 20). The mechanistic model, termed the reduced-resistances model, demonstrates that a solubilizing receiver reduces diffusional resistance in the membrane and in the receiver-side boundary layer. At high ratios of receiver to donor solubility, a hyperbolic relationship was observed where diffusion through the donor-side boundary layer becomes rate-limiting. Additional drug cocktail permeability studies with antipyrine, phenytoin, and meloxicam confirmed the broader applicability of this model. These findings provide a framework for informed receiver selection in permeability assays and underscore the importance of considering the receiver medium when comparing results across experiments.

The limited penetration of nanomedicines into tumor tissues remains a major obstacle to their therapeutic efficacy. To overcome this barrier, we designed a novel nanodrug that leverages receptor/cation dual pathway-mediated transcytosis to achieve deep tumor penetration and targeted disruption of mitochondria, resulting in significantly enhanced antitumor outcomes. The multifunctional carrier, P1, was constructed through the synthesis of an amphiphilic block copolymer, terminal conjugation of the CRGDK peptide, and side-chain modification with 7-diethylaminocoumarin (DEAC) for mitochondrial targeting. Dynamic light scattering analyses confirmed the pH/ROS-responsive behavior of P1 micelles, including acid-triggered charge reversal. Drug release kinetics, cellular uptake and endocytic mechanisms, lysosomal escape efficiency, mitochondrial colocalization, induction of ROS generation, mitochondrial membrane potential (ΔΨ_m) depolarization, apoptosis induction, penetration in multicellular tumor spheroids (MTSs) and in vivo tumors, and antitumor efficacy (in vitro and in vivo) were systematically evaluated. Results indicated that DOX/P1 micelles initially target tumor tissue via CRGDK binding, followed by NRP-1-mediated transcytosis. Subsequent acidity-induced charge reversal activates a secondary cation-mediated transcytosis pathway, synergistically promoting deep tumor infiltration. Upon mitochondrial localization, the carrier undergoes ROS-triggered degradation, leading to concurrent release of doxorubicin (DOX) and cinnamaldehyde (CA) within mitochondria. This dual release acts synergistically to amplify oxidative stress, collapse ΔΨ_m, and induce mitochondrial DNA damage, collectively precipitating irreversible apoptosis. This study establishes a programmable platform for developing tumor-penetrating nanotherapeutics with precise subcellular organelle-targeting capabilities.

PRMT5, a member of the arginine methyltransferase family, is mainly distributed in the nucleus and cytoplasm and is closely involved in tumorigenesis, development, and metastasis. Based on GSK3326595, we first designed and synthesized a PRMT5-targeted precursor (DOTA-FZ-P5R). MOE simulation results indicated a strong binding affinity (KD ≈ 10^–11) of DOTA-FZ-P5R toward PRMT5 (PDB: 4 × 61). The ⁶⁸Ga-labeled tracer exhibited high RCP (>98%) and excellent stability. Western blot and cell uptake studies confirmed that PRMT5 was highly expressed in MDA-MB-231 and AsPC1 cells, while expression in A549 cells was comparatively low. Correspondingly, microPET-CT imaging demonstrated a significantly higher uptake of [⁶⁸Ga]Ga-DOTA-FZ-P5R in MDA-MB-231 and AsPC1 tumor-bearing mice compared to A549. At 30 min postinjection, uptake values were 3.47 ± 1.53%ID/g for MDA-MB-231, 3.63 ± 0.81%ID/g for AsPC1, and 1.53 ± 0.25%ID/g for A549. These results were consistent with PRMT5 expression levels confirmed by IHC, further validating the tracer’s specificity and potential for imaging PRMT5 expression in vivo. [⁶⁸Ga]Ga-DOTA-FZ-P5R can dynamically visualize and quantify PRMT5 expression levels in real time across pan-cancer. This research demonstrates that [⁶⁸Ga]Ga-DOTA-FZ-P5R enables rapid imaging of PRMT5-positive tumors. The probe has significant potential to enable individualized and precise diagnosis in patients with PRMT5-positive tumors, define an optimal treatment window, assess therapeutic efficacy, and serve as a predictive imaging modality for tumor resistance.

Glioblastoma multiforme (GBM) is highly angiogenic, which promotes its growth and invasion. Photodynamic effects not only kill tumor cells but also disrupt or seal the tumor blood vessels. Targeting strategies that integrate effective tumor photodynamic therapy (PDT) with imaging-based vessel monitoring could lead to improvement in the diagnosis and treatment of GBM. Herein, we developed a biomimetic photosensitizer using a macrophage membrane hybrid lipid as the carrier of chlorin e6 to form a nanocomposite (MLCNPs). The MLCNPs demonstrated good biocompatibility and targeted GBM cells. Under laser irradiation, the MLCNPs showed significant photodynamic effects, which induced massive cell apoptosis. The targeting ability and PDT effect were investigated in an orthotopic glioma mouse model. During PDT in vivo, high-resolution photoacoustic (PA) imaging was used to monitor changes in the structure and function of the tumor vasculature. MLCNPs combined with high-resolution PA imaging provide a new strategy for GBM tumor diagnosis, treatment, and monitoring.

Pancreatic cancer presents significant imaging challenges due to its poor vascularization, while the hypoxic tumor microenvironment further contributes to chemoresistance. To address these limitations, we engineered exosome-ultrasmall iron oxide (Exo-USIO), a targeted exosomal nanoprobe encapsulating USIO nanoparticles (USIO NPs), designed to enable precise tumor imaging and enhance chemotherapy efficacy in pancreatic cancer. Exosomes derived from Panc-02 pancreatic cancer cells were isolated and loaded with USIO NPs via electroporation to synthesize Exo-USIO. The nanoprobe’s targeting specificity, MRI contrast enhancement, and catalase-like activity (converting H₂O₂ to O₂) were systematically evaluated. In vitro assays assessed cellular uptake, hypoxia modulation, and chemosensitivity, while in vivo studies validated tumor-targeted MRI imaging, hypoxia alleviation, and synergistic therapeutic effects with gemcitabine (GEM). Exo-USIO demonstrated a 2.3-fold increase in T₁-weighted MRI signal intensity compared to free USIO NPs (P < 0.01), alongside efficient enzymatic conversion of H₂O₂ to O₂, significantly reducing HIF-1α expression (P < 0.05). Combined with GEM, Exo-USIO reduced tumor cell viability to 39.8% in vitro and suppressed tumor growth by 62% in vivo (P < 0.001). Biosafety evaluations revealed negligible systemic toxicity or metastatic risk. By leveraging exosome-mediated targeted delivery and the dual enzyme-mimetic activity of USIO NPs, Exo-USIO achieves dual functionality: enhanced MRI-guided tumor localization and catalytic alleviation of hypoxia to reverse chemoresistance. This strategy overcomes key limitations of the pancreatic tumor microenvironment, offering a translatable platform for precision theranostics.

As an emerging modality for treatment, photothermal therapy demonstrates significant potential for clinical application. However, the inflammatory reaction after photothermal therapy can lead to tumor recurrence and metastasis. As a novel photothermal agent, biliverdin (BV) also demonstrates a remarkable anti-inflammatory effect. In this study, goat milk-derived extracellular vesicles (GEVs) is used to encapsulate BV. The objective was to enhance tumor uptake of the photothermal agent while alleviating the inflammatory responses associated with photothermal therapy, thereby achieving superior therapeutic outcomes. N₃-GEV@BV was successfully synthesized. Additionally, it exhibited notable efficacy in photothermal therapy and demonstrated anti-inflammatory effects in vitro. Utilizing a pretargeting strategy, N₃-GEV@BV can accomplish PET/CT imaging in both subcutaneous and orthotopic tumor models. After photothermal treatment, the tumor volume in the N₃-GEV@BV+laser group exhibited a significant decrease relative to the other groups, with reductions of up to 1/13 observed. Furthermore, compared to N₃-GEV@ICG, mice injected with N₃-GEV@BV exhibited lower expression levels of inflammatory factors in both the serum and tumor tissues. As an integrated nanoprobe for diagnosis and treatment, N₃-GEV@BV can successfully mediate the photothermal therapy of tumor tissue. Notably, it contributes to enhanced tumor prognosis by mitigating the inflammatory response induced by photothermal therapy, underscoring its broad potential for application.

Pharmaceutical cocrystallization is a promising strategy to enhance the solubility and bioavailability of hydrophobic active pharmaceutical ingredients (APIs). However, when API–coformer cocrystals exhibit incongruent melting, understanding and predicting their solubility in water becomes significantly more complex. In this work, a combined experimental and thermodynamic modeling approach is presented to investigate the API solubility enhancement in a ternary API–coformer–water system. Hydrochlorothiazide (HCT), a biopharmaceutics classification system (BCS) class IV diuretic, and nicotinamide (Nic), a generally recognized as safe (GRAS)-listed coformer, were selected as a representative system that forms an incongruently melting 1:1 cocrystal, which was confirmed experimentally using differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD). Binary solid–liquid equilibrium (SLE) data for the HCT–Nic, HCT–water, and Nic–water systems were experimentally measured at different temperatures. The nonrandom two-liquid (NRTL) model was then used to regress the binary interaction parameters from the binary SLE data. These parameters were then used to predict ternary SLE phase diagrams of the HCT–Nic–water system at 310.15 K, 330.15 K, and 350.15 K. The NRTL-modeled SLE diagrams revealed the key features of the ternary system, including the absence of cocrystal formation at 310.15 K and the emergence of a cocrystal phase region with incongruent dissolution behavior at elevated temperatures. The highest HCT solubility was obtained at the ternary API-rich eutectic composition, with a solubility enhancement factor (Φ) of 2.1–2.4 across the studied temperatures. In contrast, dissolving the 1:1 cocrystal directly in water yielded significantly lower solubility enhancements (Φ ≈ 1.0–1.3). These findings clearly demonstrate that selecting a binary HCT–Nic mixture that, upon dilution in water, reaches the eutectic composition in the ternary HCT–Nic–water system yields greater solubility enhancement than starting from the cocrystal composition. This study emphasizes the importance of thermodynamic modeling in understanding solubility behavior and guiding the rational design of cocrystal-based pharmaceutical formulations, especially for API–coformer systems exhibiting incongruent melting.