Molecular Pharmaceutics最新文献

Trop2 exhibits significantly elevated expression in numerous solid malignancies, playing a crucial role in tumor advancement, whereas its presence in healthy tissues is minimal. In this study, we investigated Trop2 expression in bladder cancer models using [⁶⁴Cu]Cu-NOTA-Trodelvy for immunoPET imaging. In HT-1376 models, [⁶⁴Cu]Cu-NOTA-Trodelvy effectively visualized tumor as early as 12 h p.i. (10.30 ± 1.45% ID/g), with tumor uptake increasing and peaking at 48 h p.i. (13.73 ± 1.16% ID/g), highlighting its potential for tumor imaging. Control groups also demonstrated low tumor uptake (5.27 ± 1.14% ID/g at 48 h in the blocking group; 6.33 ± 0.74% ID/g at 48 h in UM-UC-3; 4.50 ± 0.30% ID/g at 48 h in the [⁶⁴Cu]Cu-NOTA-IgG group). Long-term fluorescence imaging further confirmed the tumor uptake rate in the IRDye 800CW-Trodelvy group was significantly higher than in the IRDye 800CW-Trodelvy blockade group (P < 0.001). Our findings demonstrated that [⁶⁴Cu]Cu-NOTA-Trodelvy enables specific and prolonged tumor accumulation in bladder cancer models, providing precise and noninvasive monitoring of Trop2 expression.

As the major driving factor of hepatic fibrosis, the activated hepatic stellate cells (aHSCs) rely on active glycolysis to support their aberrant proliferation and secretion of the extracellular matrix. Sorafenib (Sor) can combat liver fibrosis by suppressing HIF-1α and glycolysis, but its poor solubility, rapid metabolism, and low bioavailability restrict such a clinical application. Here, Sor was loaded onto polydopamine nanoparticles and then encapsulated by a retinoid-decorated red blood cell membrane, yielding HSC-targeted Sor nanovesicles (PDA/Sor@RMV-VA) with a high Sor-loading capacity and photothermally controlled drug release for antifibrotic treatment. These Sor RMVs not only exhibited a good particle size, dispersity and biocompatibility, prolonged circulation time, enhanced aHSC targetability, and hepatic accumulation both in vitro and in vivo, but also displayed a mild photothermal activity proper for promoting sorafenib release and accumulation in CCl₄-induced fibrotic mouse livers without incurring phototoxicity. Compared with nontargeting Sor formulations, PDA/Sor@RMV-VA more effectively downregulated HIF-1α and glycolytic enzyme in both cultured aHSCs and fibrotic mice and reversed myofibroblast phenotype and amplification of aHSCs and thus more significantly improved liver damage, inflammation, and fibrosis, all of which could be even further advanced with NIR irradiation. These results fully demonstrate the antifibrotic power and therapeutic potential of PDA/Sor@RMV-VA as an antifibrotic nanomedicine, which would support a new clinical treatment for hepatic fibrosis.

The optimization of absorption, distribution, metabolism, and excretion (ADME) profiles of compounds is critical to the drug discovery process. As such, machine learning (ML) models for ADME are widely used for prioritizing the design and synthesis of compounds. The effectiveness of ML models for ADME depends on the availability of high-quality experimental data for a diverse set of compounds that is relevant to the emerging chemical space being explored by the drug discovery teams. To that end, ADME data sets from Genentech and Roche were combined to evaluate the impact of expanding the chemical space on the performance of ML models, a first experiment of its kind for large-scale, historical ADME data sets. The combined ADME data set consisted of over 1 million individual measurements distributed across 11 assay end points. We utilized a multitask (MT) neural network architecture that enables the modeling of multiple end points simultaneously and thereby exploits information transfer between interconnected ADME end points. Both single- and cross-site MT models were trained and compared against single-site, single-task baseline models. Given the differences in assay protocols across the two sites, the data for corresponding end points across sites were modeled as separate tasks. Models were evaluated against test sets representing varying degrees of extrapolation difficulty, including cluster-based, temporal, and external test sets. We found that cross-site MT models appeared to provide a greater generalization capacity compared to single-site models. The performance improvement of the cross-site MT models was more pronounced for the relatively “distant” external and temporal test sets, suggesting an expanded applicability domain. The data exchange exercise described here demonstrates the value of expanding the learning from ADME data from multiple sources without the need to aggregate such data when the experimental methods are disparate.

Photothermal therapy (PTT) uses near-infrared (NIR) light and a photothermal agent (PTA) to generate heat to kill tumor cells. PTT is an attractive therapy for highly metastatic tumors─such as triple-negative breast cancer (TNBC)─because PTT is a potent activator of immunogenic cell death (ICD). ICD is characterized by the production of damage-associated molecular patterns (DAMPs) that help the immune system recognize cancer cells as “nonself.” This generates an immune response against the tumor cells and helps to combat both primary and metastatic tumors. However, an unknown thermal window remains in which ICD is most prevalent. Here, we conjugate an NIR-absorbing dye to the surface of bacteriophage Qβ to generate a viral-based PTA. Additionally, we demonstrate that mild PTT (<45 °C) is not enough to cause significant apoptosis in the murine TNBC model. In comparison, hot PTT (>60 °C) effectively eliminates cancer cells but is less likely to induce ICD. An optimal temperature range is moderate PTT (50–60 °C), where effective cell killing and ICD occur. We show an increased surface expression of DAMPs within this range, along with an increased ratio of pro- to anti-inflammatory cytokines by dendritic cells.

Poor stability of biological products such as proteins is a major challenge facing the biopharmaceutical industry. Poor stability is usually mitigated by formulating these products in the solid state, employing sugars as stabilizers. Several studies have pointed out the superior stabilizing ability of disaccharides, including sucrose and trehalose, as compared to polysaccharides such as dextrans. The aim of this study was to investigate the impact of excipient molecular weight on miscibility with a model protein, bovine serum albumin (BSA). Aqueous solutions containing a binary combination of a sugar-based stabilizer and BSA were dried using different methods (air drying to form films, spray drying, and lyophilization). The stabilizers tested varied in molecular weight and were dextran 6, 70, or 2000 kDa, hydroxypropyl methyl cellulose (HPMC), and trehalose. Miscibility was evaluated using a variety of techniques including confocal fluorescence microcopy, infrared and Raman microscopy, and solid-state nuclear magnetic resonance (ssNMR) spectroscopy. The stability of BSA in dried mixtures subjected to accelerated storage conditions was also measured. BSA was more stable in the presence of dextran 2000 kDa compared to dextran 70 and 6 kDa, while stability was highest in trehalose and lowest in HPMC. From ssNMR spectroscopy, BSA-Dex 2000 kDa and BSA-trehalose were miscible over 20 and 5 nm length scales, BSA-Dex 6 kDa was miscible over a 20 nm length scale and phase-separated over a 5 nm length scale, while BSA-Dex 70 kDa and BSA-HPMC were phase-separated over both length scales. It was postulated that for dextran, the size of the polysaccharide relative to the size of the protein determined the extent of the system miscibility and stability. A smaller or similar polysaccharide size compared to that of the protein, as in the case of BSA-Dex 6 kDa and BSA-Dex 70 kDa, leads to depletion-induced phase separation. A much larger polysaccharide size compared to that of the protein allows the protein molecules to be trapped within a polysaccharide mesh, resulting in a miscible system. This study suggests that the impact of the relative size of the stabilizer and protein on miscibility is more complex than previously considered.

Many new drug substances exhibit poor physicochemical properties and therefore require significant time and material resources to develop into safe and efficacious medicinal products. This typically involves exploring a large amount of compositional space and may require excessive amounts of drug compounds, which may not be adequate at the early stage of drug development. Scaled-down screening methods have been used as a cost-effective approach to the early-stage formulation. However, even the most material-efficient methods used in product development require milligrams or grams of drug material, which is often not available until relatively late in the lead optimization process. Herein, we report the application of picoliter inkjet printing of drugs and polymers from solution to create addressable formulation microarrays. This allows the efficient screening of drug–polymer compositions while only requiring micrograms or less of the drug substance. A total of eight model compounds, namely, carbamazepine, griseofulvin, saccharin, theophylline, 4-aminobenzoic acid, caffeine, salicylic acid, and benzocaine, were screened against seven commonly used amorphous solid dispersion (ASD) matrix polymers at 5% w/w composition intervals in the range of 5–80% w/w, with five replicates each. Each dispensed spot contains a total of only 1 μg of material (model compound and/or polymer). Across the tested ASD formulations, we ranked the different polymers based on their ability to hinder drug recrystallization across different compositions. Also, we identified distinct physicochemical behaviors in their crystallization kinetics, such as moisture resolubilization. We expect this approach to enable the rapid time- and material-efficient development of new amorphous solid dispersion formulations in an industrial setting.

The microspecies-specific physicochemical properties of eutectic solids of sparingly water-soluble micronutrient, Vitamin B₂ (Riboflavin), with a few representative BCS drugs, viz., Theophylline, Theobromine, Mesalamine, and Barbituric acid are established. The interplay of solubility and drug permeation behavior is experimentally determined for the eutectic solids, and properties are corroborated with the concomitant relative concentrations of pH-dependent microspecies of Riboflavin and the drugs. Partner drug candidates are selected from different quadrants of BCS classification to apprehend the influence of their solubility on the overall efficacy of the eutectic solids. The coexistence and inseparable ionic, neutral, and/or zwitterionic microspecies are spotted, and the pH-reliant isomer-specific inflection of physicochemical and pharmacokinetic properties in such multicomponent solid formulations is demonstrated.

Structural studies of pharmaceutical hydrates are essential to understanding stability-related issues, especially during the heating process of formulation. A thorough understanding of the hydration and dehydration behavior of active pharmaceutical ingredient (API) hydrate is also important since phase transitions can occur during the formulation process. This is because dehydration could result in a considerable rearrangement in the structure if water–API hydrogen bonding is present. We perform advanced solid-state NMR experiments on regorafenib monohydrate to investigate the role of fluorine in hydrogen bonding interaction, and the results are compared to its anhydrous form and its structural analogue, namely, sorafenib. Our results show that significant structural changes could not be observed on dehydration. Based on our study, it can be concluded that the introduction of fluorine restricts the intramolecular hydrogen bonding and the asymmetry in the structure of regorafenib monohydrate is absent, in comparison to sorafenib.

The engineerability of lipid nanoparticles (LNPs) and their ability to deliver nucleic acids make LNPs attractive tools for cancer immunotherapy. LNP-based gene delivery can be employed for various approaches in cancer immunotherapy, including encoding tumor-associated antigens and silencing of negative immune checkpoint proteins. For example, LNPs carrying small interfering RNAs can offer several advantages, including sustained and durable inhibition of an immune checkpoint protein. Due to their tunable design, modifying the lipid composition of LNPs can regulate the rate of their uptake by immune cells and the rate of gene silencing. Controlling the kinetics of LNP uptake provides additional flexibility and strategies to generate appropriate immunomodulation in the tumor microenvironment. Here, we evaluated the effects of polyethylene glycol (PEG) content ranging from 0.5 to 6 mol % on the cellular uptake of LNPs by immune cells and gene silencing of PD-L1 after intratumoral administration. We evaluated the cellular uptake and PD-L1 blockade in vitro in cell studies and in vivo using the YUMM1.7 melanoma tumor model. Cell studies showed that the rate of cell uptake was inversely correlated to an increasing mol % of PEG in a linear relationship. In the in vivo studies, 0.5% PEG LNP initiated an immediate effect in the tumor with a significant decrease in the PD-L1 expression of immune cells observed within 24 h. In comparison, the gene silencing effect of 6% PEG LNP was delayed, with a significant decrease of PD-L1 expression in immune cell subsets being observed 72 h after administration. Notably, performance of the 6% PEG LNP at 72 h was comparable to that of the 0.5% PEG LNP at 24 h. Overall, this study suggests that PEG modifications and intratumoral administration of LNPs can be a promising strategy for an effective antitumor immune response.