Molecular Pharmaceutics最新文献

Effective management of solid tumors requires both surgical resection and sustained local chemotherapy. However, immediate postoperative drug delivery remains challenging due to bleeding risk and the delay in systemic therapy. Here, we report a coassembled peptide/doxorubicin hydrogel (BDO) designed to address this clinical gap by integrating intraoperative hemostasis with localized chemotherapy. The hydrogel is formed via coassembly of a functional peptide (B-cL1), which combines VEGFR inhibition and nanofiber-forming capacity, with doxorubicin. During surgery, the hydrogel rapidly adheres to the tissue interface and promotes hemostasis while enabling sustained drug release at the tumor resection site. Structural characterization and molecular simulations confirm a stable fibrous network facilitated by hydrogen bonding and hydrophobic interactions. In vitro and in vivo studies demonstrate excellent biointerface compatibility, enhanced coagulation, and localized antitumor activity with minimal systemic toxicity. These findings highlight a promising biointerface-engineered strategy for simultaneous bleeding control and site-specific chemotherapeutic delivery during tumor surgery.

Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers due to late-stage diagnosis, aggressive progression, inadequate modalities for detection and monitoring, and treatment options, including surgery, that generally do not achieve durable responses. CUB domain containing protein 1 (CDCP1) and mesothelin (MSLN) are cell surface proteins, commonly expressed at elevated levels in PDAC, that are potential targets for detection and treatment of these tumors. In this study, we generated anti-CDCP1 antibody ch10D7 and anti-MSLN antibody amatuximab conjugated with the near-infrared fluorophore indocyanine green (ICG). With the goal of characterizing ch10D7^ICG and anti-MSLN^ICG for detection of PDAC, we noted that ICG labeling did not impact the affinity or specificity of either antibody. Both ICG-labeled antibodies selectively accumulated in subcutaneous and orthotopic PDAC models in mice, as visualized by in vivo fluorescence imaging. Postmortem fluorescence endoscopy imaging clearly delineated tumors, with optimal signal observed at 120 h post agent administration. Demonstrating specificity in vivo, depletion of CDCP1 and MSLN abolished tumor localization in xenografts of, respectively, ch10D7^ICG and amatuximab^ICG. Quantitative ex vivo fluorescence analysis of excised xenografts demonstrated that combining ch10D7^ICG and amatuximab^ICG enhances tumor-associated fluorescence by 2.8- to 12.5-fold compared with either agent alone. The results support the potential of dual-targeted, antibody-based fluorescence imaging for enhanced intraoperative visualization and excision of PDAC, including for tumors with heterogeneous expression of either or both of the targeted receptors CDCP1 and MSLN.

As the diversity of therapeutic protein structures continues to evolve, it is essential to understand the mechanisms that determine their pharmacokinetic properties. The current work was initiated to establish the physicochemical attributes and cellular processes most crucial for the target-independent disposition of proteins possessing a fragment crystallizable (Fc) region. We systematically redesigned the surface properties of five de novo-generated protein scaffolds lacking any known binding partner in mice to produce a total of 35 Fc-fused proteins exhibiting a diverse set of physicochemical characteristics. Pharmacokinetic studies in wild-type mice revealed a profound spread in elimination rates and extensive tissue accumulation that was most strongly associated with charge descriptors. A suite of in vitro studies demonstrated that these in vivo observations significantly correlated to cellular nonspecificity wherein positive surface charge caused higher nonspecific adsorptive endocytosis, diminished recycling efficiency by the neonatal Fc receptor, and net cellular accumulation. Combined, our results provide a detailed explanation for how the disposition of Fc-fused proteins is impacted by charge, which will aid protein engineering efforts aimed at optimizing pharmacokinetic features.

Gallium maltolate (GaM) targets iron-dependent processes in glioblastoma (GBM), but responses vary with the model context. We evaluated GaM across established (A-172, U-87 MG) and patient-derived (3005, 3019, 3034, 3048, 3073) GBM lines in 2D and 3D using viability modeling (IC10/IC50/IC90), transferrin receptor (TFRC) quantification, oxygen consumption rate (OCR), and PCA/PLS-DA-guided metabolomics with false discovery rate (FDR) and variable importance in projection (VIP)-based selection. GaM reduced viability in all models, but the impact of 3D culture on IC50 was line-specific rather than uniformly increasing resistance: classical/proneural patient-derived lines (3005, 3019, 3048) showed equal or lower IC50 in 3D compared with 2D, 3073 showed minimal change, whereas the mesenchymal-like line 3034 displayed a marked IC50 increase in 3D. Basal TFRC levels correlated with IC50 in 2D but not 3D, indicating that the TFRC alone does not predict GaM response once microenvironmental constraints are introduced. Instead, a broader phenotype involving TFRC/CD44/MGMT and TFR2 expression is associated with 3D sensitization versus protection. OCR was markedly suppressed in A-172, U-87 MG, 3048, and 3073, particularly in 3D, while 3005 and 3019 were more respiration-resilient. Multivariate analyses showed treatment-dominant separation in 3005 and 3048, format dominance in 3019 and 3034, and time effects in A-172/U-87 MG/3073. A concise metabolic signature consisting of tryptophan, methionine, uracil, and allantoin indicated coordinated perturbations in amino acid, nucleotide, and redox pathways. These findings support complementary 2D and 3D patient-derived GBM models for mechanistic studies and the predictive evaluation of GaM.

Nanotheranostics have brought a new era in cancer treatment. However, obstacles to their wider use persist in the areas of biodistribution, safety profiling, and clinical translation. Consequently, for nanotechnology to work properly, it is crucial to assess its safety and potential therapeutic applications. However, the traditional mammalian models face constraints due to their high cost, ethical concerns, and low throughput. An effective substitute to overcome these restrictions in nanotheranostics research are the xenotransplant, genetic, and chemically induced zebrafish (Danio rerio) models. The zebrafish offers several benefits as a model organism for research into cancer treatments. Due to their short life cycle, high degree of genetic resemblance to humans, and well-studied organ systems, they are an excellent candidate for pharmacokinetic and toxicological studies. Factors including complicated pharmacokinetics, organ-specific toxicity, and unexpected in vivo behavior frequently impede the progress of nanotheranostics for biomedical uses, especially in cancer treatment and drug delivery. To overcome the aforementioned limitations, various nanotheranostics such as liposomes, mesoporous silica nanoparticles (NPs), magnetic NPs, exosomes, micelles, polymerosomes, etc., showed a promising outcome in enhancing drug delivery, improving therapeutic efficacy, and reducing systemic toxicity. This review discusses the utility of zebrafish larvae cancer models in evaluating nanotheranostics, with emphasis on factors influencing nanoparticle biodistribution and the potential of targeted drug delivery within these models.

Breast cancer remains one of the most prevalent cancers among women, with triple-negative breast cancer (TNBC), lacking estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2, accounting for approximately 15–20% of all patients with breast cancer. TNBC is notably aggressive, with a high invasive, metastatic, and recurrence potential. In this study, we found that the migration and invasion capabilities of MDA-MB-231 cells, derived from human TNBC, were strongly influenced by the serum concentration. Transwell assays revealed that TNBC cell migration varied depending on the fetal bovine serum (FBS) level, with an optimal concentration that substantially enhanced migration and invasion. In contrast, non-TNBC MCF-7 cells exhibited no such serum-dependent migration pattern. In addition to data-independent acquisition (DIA) phosphoproteomic analysis for understanding the mechanisms, the cellular uptake of the flock house virus coat (35–49) peptide, a type of arginine-rich cell-penetrating peptide with serum-dependent cellular uptake efficacy, was significantly increased under the optimal serum conditions, which induces cell migration, leading to efficient delivery of apoptosis-inducible peptide and TNBC-killing activity. Our findings highlight the critical role of serum concentration in regulating TNBC behavior and offer insights into leveraging serum-responsive delivery systems for targeted breast cancer therapy.

Nanobody molecules are single-domain antibodies derived from stable and fully functional heavy-chain-only (VHH) camelid antibodies. Due to their small size, approximately 15 kDa, their tissue distribution differs from that of conventional antibodies. This study investigates the whole-body pharmacokinetics (WBPK) and specific binding of half-life extended VHHs (HLE-VHHs) using dynamic whole-body immunoPET imaging in nonhuman primates. A HLE-VHH targeting the IL-6 receptor (CD126-VHH) and its nontargeting control (IRR-VHH) were evaluated in vivo after zirconium-89 radiolabeling. We compared microdoses (0.18 ± 0.11 mg/kg, n = 2) and/or pharmacological doses (coinjection, 3.19 ± 0.02 mg/kg, n = 2) of IRR-VHH, alongside microdoses of CD126-VHH (0.25 ± 0.14 mg/kg, n = 2). Initial dynamic whole-body PET scans (240 min) were followed by static PET scans on days 1, 7, and 14. We validated an image-derived input function against the arterial blood sampling method for kinetic modeling. [⁸⁹Zr]IRR-VHHs showed predominantly vascular distribution from day 0 to day 14, with comparable activity patterns and tissue distribution between microdose and pharmacological dose administrations. Early [⁸⁹Zr]CD126-VHH distribution mirrored the nonspecific IRR-VHH pattern, showing strong correlation (R² = 0.87, slope = 1.1) in uptakes of organs, normalized to blood (standard uptake value ratio, SUVR) values at day 0. Interestingly, by days 7 and 14, [⁸⁹Zr]CD126-VHH exhibited higher uptake in bone marrow, spleen, liver, and kidneys, suggesting specific binding. The distribution of [⁸⁹Zr]IRR-VHH in these organs was assumed to reflect the nonspecific uptake of VHHs and was used to estimate the specific binding of [⁸⁹Zr]CD126-VHH. The specific binding of [⁸⁹Zr]CD126-VHH was similarly estimated using either the arterial input function or the image-derived input function (R² = 0.997, p < 0.0001). In plasma, the shorter elimination half-life of [⁸⁹Zr]CD126-VHH (2.0–1.8 days) compared to microdose [⁸⁹Zr]IRR-VHH (5.1–5.0 days) or pharmaco-dose [⁸⁹Zr]IRR-VHH (4.9–4.8 days) suggested target-mediated drug disposition. This study demonstrates the potential of immunoPET for evaluating the target-mediated drug disposition and WBPK of VHH molecules in vivo, providing valuable insights into VHH-based therapeutic strategies.

⁹⁰Y-microsphere radioembolization is an established treatment for primary and metastatic liver cancer. Post-treatment PET/CT enables voxel-level absorbed dose estimation, but the optimal calculation method remains under evaluation. In this study, we compared three dosimetry approaches: Monte Carlo (MC), voxel S-value (VSV) method, and local energy deposition method (LDM). Post-treatment PET/CT data sets from 68 patients were analyzed (6 from the Deep Blue Data Repository and 62 from Beijing Tsinghua Changgung Hospital). MC dosimetry was implemented in GATE v9.0 and served as the reference standard. VSV was derived by convolving PET images with a precomputed ⁹⁰Y voxel kernel, while LDM assumed complete local absorption of β-energy. Agreement with MC was assessed qualitatively using relative-difference maps, dose profiles, and isodose overlays, and quantitatively via voxelwise root-mean-square error (RMSE), Pearson correlation, Bland–Altman analysis of mean absorbed dose and equivalent uniform biological effective dose (EUBED), and absolute maximum deviations between cumulative and differential dose-volume histograms (DVHs) (1 Gy bins). MC simulations required ∼41 h per case, whereas the VSV kernel calculation required only one simulation with ∼6 h. Then VSV and LDM costed only a few seconds to obtain the dose images. VSV preserved MC isodose geometry and dose-profile shapes, while LDM overestimated doses in high-dose regions. At profile maxima, VSV’s peak relative deviation was ∼1–3% versus ∼12–19% for LDM. Across patients, VSV achieved a voxelwise RMSE of 2.21% and <3% differences in mean dose and EUBED, with tighter Bland–Altman limits than LDM. LDM showed larger bias, wider limits, and greater DVH deviations at higher doses, confirming that VSV attains near-MC accuracy with orders-of-magnitude faster computation. Overall, all three methods enable voxel-level dosimetry of ⁹⁰Y-microsphere therapy, but VSV combines MC-like accuracy with dramatically reduced computation time. VSV thus offers a clinically practical solution for rapid and reliable post-treatment dose verification and may support more personalized treatment evaluation in ⁹⁰Y radioembolization.

Developing solid dosage forms containing pharmaceutical cocrystals through a one-step process remains a major challenge in drug delivery. Here, we present a hybrid electrospinning–electrospraying strategy for fabricating indomethacin–saccharin (IND–SAC) cocrystal-loaded nanofibers, specifically engineered for ocular application. Poly(vinyl alcohol) (PVA) and poloxamer P407 were electrospun to form nanofiber scaffolds, followed by electrospray deposition of IND–SAC cocrystals onto the fiber surface. This dual fabrication approach aimed to enhance the drug solubility, enable in situ cocrystal incorporation, and achieve sustained release. The nanofibers were evaluated for rheological and mucoadhesive properties, demonstrating favorable shear thinning behavior and strong adhesion. IND–SAC cocrystals were characterized by scanning electron microscopy, differential scanning calorimetry, X-ray diffraction, Fourier-transform infrared spectroscopy, and dynamic light scattering, confirming the crystalline integrity, morphological uniformity, and absence of undesirable physicochemical interactions. In vitro release studies compared single-layer and bilayer nanofiber designs. Both single-layer and bilayer designs showed comparable 24 h release profiles which followed Higuchi diffusion kinetics in both systems. Mechanistic differences were observed (n = 0.52 vs 0.97, respectively), but these did not translate into significant differences in cumulative release. Overall, this work highlights the promise of hybrid electrospinning–electrospraying as a scalable, single-step platform for manufacturing cocrystal-loaded nanofibers. The approach combines solubility enhancement, mucoadhesion, and controlled release, addressing critical challenges in ophthalmic drug delivery and offering potential for translation into patient-friendly ocular therapies.