BackgroundMicroglia drive neuroinflammation in Alzheimer's disease (AD), yet no approved therapy targets this compartment. Human genome-wide association studies consistently implicate innate immune loci in AD risk, establishing microglial transcriptional programs as therapeutically relevant but pharmacologically underexploited targets.ObjectiveWe sought to identify transcription factors (TFs) governing microglial state transitions computationally and to nominate structurally tractable drug repurposing candidates.MethodsWe applied trajectory inference (PAGA), pseudobulk DESeq2, pySCENIC gene regulatory network (GRN) inference, CellChat, and virtual screening of 1962 approved compounds to 236,002 microglial nuclei from 84 donors (SEA-AD atlas).ResultsIKZF1 was the sole target TF retained under cisTarget v10 motif constraints, with peak regulon activity in LateAD-DAM (pseudotime ρ = + 0.309) and replication in an independent bulk cohort (GSE95587; adjusted p = 0.004). CellChat identified SLIT2→ROBO2 from multiple neuron subtypes (predominantly inhibitory interneurons) as the top predicted pathway to microglia. Tafamidis (→IRF8) and diflunisal (→PPARG) were top virtual screening hits; all evaluated compounds failed the pre-specified selectivity threshold.ConclusionsIKZF1 is prioritized as a candidate late-disease microglial TF, supported by six convergent evidence dimensions including independent bulk replication. Tafamidis and diflunisal are low-confidence repurposing hypotheses requiring experimental validation.

Cancer stem cells (CSCs) drive metastasis and relapse and are resistant to conventional apoptosis-inducing chemotherapies. Methuosis, a non-apoptotic form of cell death, is largely unexplored for anti-CSC therapy. Here we report a cobalt(III)-cyclam complex (1) incorporating pyrene and diflunisal moieties that induces features consistent with methuosis in a CSC model. Mechanistic investigations revealed that 1 induces phenotypes consistent with methuosis in breast CSCs, including extensive cytoplasmic vacuolation, elevated Rab7 and LAMP1 expression, activation of JNK and p38 MAPK pathways, G2/M cell-cycle arrest, augmented intracellular reactive oxygen species levels, and disruption of the mitochondrial membrane potential. Beyond direct cytotoxicity, 1 induces an anti-breast CSC immunogenic response by perturbing the inhibitory damage-associated molecular patterns axis through cyclooxygenase-2 downregulation. When formulated as polymeric nanoparticles and evaluated in a murine metastatic triple-negative breast cancer model, 1 penetrates tumour tissue and suppresses tumour growth more than cisplatin. The nanoparticle formulation activates anti-tumour immunity, characterised by enhanced infiltration of T cells and mature dendritic cells in tumours and tumour-draining lymph nodes. Intratumoral T-cell recruitment is associated with M2-to-M1 macrophage polarisation and depletion of myeloid-derived suppressor cells. This work identifies 1 as the first metal complex shown to induce features consistent with methuosis in a CSC model.

Process analytical technology (PAT) plays an essential role in pharmaceutical manufacturing by enabling effective quality control and improving production efficiency. Terahertz (THz) spectroscopy has attracted considerable attention due to its advantages of high biosafety, distinctive spectral fingerprints, and excellent quantitative performance. However, achieving rapid classification and accurate quantification of pharmaceutical content remains challenging. Therefore, the combination of THz spectroscopy and machine learning algorithms is introduced to address this challenge. Three active pharmaceutical ingredients (APIs), namely isoniazid, 4-aminobenzoic acid, and diflunisal, were selected, and principal component analysis (PCA) was first employed for dimensionality reduction of the THz spectral data. Subsequently, the prediction accuracy and computational efficiency were optimized using four machine learning algorithms, including support vector regression (SVR), multilayer perceptron (MLP), convolutional neural networks (CNNs) and backpropagation artificial neural network (BP-ANN). Among all models, the PCA-SVR model achieved the best prediction performance, with a coefficient of determination of 0.9990 and a root mean square error of 1.2%. In addition, the prediction time is reduced by up to 90%, which greatly improves the efficiency of data processing. The robustness of the proposed framework was further validated using a multi-component pharmaceutical system to simulate practical tablet formulations. This work demonstrates the potential of THz spectroscopy for component identification and content prediction of active pharmaceutical ingredients and sheds light on the nondestructive application for monitoring pharmaceutical quality and improving production efficiency in an online manner.

Hereditary transthyretin amyloidosis with polyneuropathy (ATTRv-PN) is an autosomal dominant disorder caused by pathogenic variants in the transthyretin (TTR) gene resulting in progressive sensorimotor neuropathy, autonomic dysfunction, and effects on other organs, particularly the heart. Historically, treatment options have been limited to liver transplantation and symptomatic management, and the disease was associated with poor prognosis and progressive disability. However, over the past decade, the therapeutic landscape has been transformed by the development of disease-modifying therapies that target the underlying pathophysiology of TTR amyloid formation. These therapies include TTR stabilizers, such as tafamidis, diflunisal, and acoramidis, as well as RNA-silencing agents, such as patisiran, vutrisiran, inotersen, and eplontersen, which reduce hepatic production of TTR and slow disease progression. Clinical trials of RNA interference therapies have demonstrated significant improvements in neuropathy impairment scores, quality of life, and functional outcomes, with acceptable safety profiles and sustained long-term benefits. Emerging gene-editing approaches, including CRISPR-based therapies like NTLA-2001, may provide a one-time treatment and long-term disease control, representing a promising future direction for managing ATTRv-PN. Despite these advances, important challenges remain. These include high treatment costs, limited global accessibility, uncertainty regarding optimal treatment selection and sequencing, and the need for long-term real-world, patient-centered outcome data. This review summarizes current disease-modifying therapies for ATTRv-PN, compares their mechanisms and clinical evidence, and discusses unmet needs and future research directions, with the aim of providing a practical and comprehensive overview for clinicians treating patients with ATTRv-PN.

The pathogenesis of hereditary ATTR (ATTRv) amyloidosis has been elucidated, and disease-modifying drugs with multiple mechanisms of action are used clinically. The International Society of Amyloidosis (ISA) guidelines strongly recommend treating ATTRv amyloidosis patients with transthyretin (TTR) tetramer stabilizers or TTR gene silencers (e.g. small-interfering RNA, antisense oligonucleotide). Tafamidis and vutrisiran are indicated for both polyneuropathy (ATTRv-PN) and cardiomyopathy (ATTR-CM), patisiran, eplontersen and diflunisal are for ATTRv-PN only, and acoramidis is for ATTR-CM only. Eligibility for treatment varies by country and is heterogeneous. These disease-modifying drugs are most effective before significant end-organ dysfunction has ensued, making early diagnosis of ATTRv amyloidosis essential. With multiple treatment options existing, it is important to accumulate evidence regarding treatment selection, switching and combination therapy. The currently available disease-modifying therapeutics have a limited effect on central nervous system (CNS) and ocular manifestations since they target either blood circulating TTR from the liver or ATTR amyloid fibrils misfolding in the blood and depositing in systemic (non-CNS, non-ocular) tissues. Development of disease-modifying therapies for CNS and ocular ATTR amyloidosis is an urgent unmet medical need.

The YAP/TAZ-TEAD complex, a core downstream co-regulatory module of the Hippo pathway, drives tumorigenesis and progression upon aberrant activation. Targeting its protein-protein interaction (PPI) is a highly promising anticancer strategy. TEAD palmitoylation maintains TEAD homeostasis and facilitates YAP/TAZ binding, and this pocket has been successfully exploited to develop covalent and non-covalent TEAD inhibitors. To discover novel chemotype TEAD inhibitors, we established a strategy integrating virtual screening, drug repurposing and structure-based optimization, and recently reported several FDA-approved drugs as potential TEAD inhibitors. Herein, we identify NSAID diflunisal (DF) as a potent TEAD inhibitor, elucidating its undefined anticancer mechanism. Structure-based optimization of DF further yielded novel biphenyl-structured covalent TEAD inhibitors, and representative derivative DF-CIV-2 features a unique sagitta-shaped binding conformation, potently inhibiting TEAD palmitoylation, transcriptional activity and YAP-TEAD complex formation, downregulating Hippo target genes, suppressing YAP-dependent cancer cell proliferation and 3D spheroid growth, and synergizing with MEK/PI3Kα/mTORC1 inhibitors. Collectively, we established a feasible TEAD inhibitor discovery pipeline, and identified DF and its sagitta-shaped derivative DF-CIV-2 as promising TEAD inhibitors, providing novel molecular tools and insights for YAP/TAZ-TEAD-targeted cancer therapy.

Transthyretin (TTR) amyloidosis is one of the most common forms of systemic amyloidosis, involving deposits of pathogenic TTR aggregates in tissues and organs throughout the body. TTR aggregation is initiated by the dissociation of a native TTR tetramer into a monomeric intermediate, which subsequently misfolds and assembles into insoluble aggregates. Using an efficient [19]F-NMR aggregation assay, we previously showed that designed peptide inhibitors can interact with multiple TTR species. However, quantifying species-specific binding affinities has remained difficult because the populations of these TTR species change over time and include a low-abundance monomeric intermediate. Here, we develop a quantitative method to extract species-dependent binding affinities directly from population-resolved [19]F-NMR aggregation data. Using diflunisal as a model compound, we determine its binding affinities for both the tetramer and the monomeric intermediate under acidic, aggregating conditions. At acidic pH, tetramer binding becomes approximately 2-fold tighter, and monomer binding becomes 15-fold stronger, compared to neutral pH. We then apply this method to previously collected [19]F-NMR aggregation data for designed peptide inhibitors. The results show that concatenating two capping peptides increases their binding affinity to both TTR tetramers and monomeric intermediates by about 2-fold, relative to the same peptides mixed separately at equal concentrations. Our method enables direct, quantitative comparison of species-dependent inhibitor binding, providing mechanistic insights useful for designing and optimizing TTR aggregation inhibitors.

The polyfluorinated biaromatic framework is a key structural motif in pharmaceutical chemistry and advanced functional materials. However, its synthesis is frequently hindered by multistep processes and stringent reaction conditions. In this study, we present a nickel-catalyzed reductive cross-coupling strategy between halogenated arenes, offering a streamlined and efficient route to polyfluorinated biaryl hydrocarbons via a radical mechanism. This method exhibits broad substrate scope and significantly reduces the complexity of synthetic routes to a range of fluorinated compounds, including liquid crystals, the nonsteroidal anti-inflammatory drug diflunisal, and aminopeptidase inhibitors, highlighting its practical relevance. Mechanistic investigations indicate that a nickel(0) species facilitates the activation of polyfluorinated aryl bromides, generating key polyfluoroaryl radicals that drive the cross-coupling process. Overall, this work introduces a practical and versatile approach to accessing polyfluorinated biaryl structures and broadens the applicability of reductive biaryl coupling methodologies.

Dendrimers (DDs) have become a pioneering tool in dermatological nanomedicine thanks to their branched nanostructure, high precision, and multifunctional nature. The purpose of this review is to highlight the revolutionary potential of DDs regarding the primary barrier for skin drug delivery - the stratum corneum. The enhanced solubility, controlled drug release, and facilitated drug penetration achieved by the customized dendrimer architecture are described in terms of diffusion, degradation, and stimuli-responsive mechanisms. The latest achievements are critically assessed through the topical approaches for psoriasis, wound healing, and skin cancer treatment, as well as transdermal delivery for systemic vaccines and gene therapy. The essential aspect of surface engineering is also analyzed in terms of its dual-edged features-chemical functionalization optimizes skin affinity and therapeutic specificity while addressing toxicological issues. Quantitative data from the literature have been provided to support the benefits, such as a 4.5-fold increase in the transdermal flux of indomethacin using PAMAM G4 and 2.7-fold and 2.5-fold increases in the bioavailability of ketoprofen and diflunisal, respectively. Moreover, in the studies applying dendrimers combined with sonophoresis, the permeability results demonstrated a noticeable trend - e.g., 935.21 μg/cm2 versus 56.69 μg/cm2 permeation, which indicates the possibility of synergism with physical enhancement methods Moreover, in the studies applying dendrimers combined with sonophoresis, the permeability results demonstrated a noticeable trend - e.g., 935.21 μg/cm2 versus 56.69 μg/cm2 permeation, which indicates the possibility of synergism with physical enhancement methods. To ensure successful transfer to clinics, an increased focus on long-term safety, synthesis scalability, and reproducibility is needed. This article provides an in-depth, objective, and holistic assessment of dendrimers as a versatile, continually expanding and improving toolkit for targeted skin therapies.

Hydrogen bonds (H-bonds) can have a critical impact on the stability and drug release characteristics of amorphous solid dispersions (ASDs). Based on the structure of the ASD components, it remains however difficult to predict the strength and influence of these H-bond interactions on the phase behavior of the ASDs. Therefore, this study assessed the miscibility and H-bond interactions of diflunisal (DIF) and four structural analogues in ASDs with Eudragit S100 (ES100) as a model polymer that contains H-bond-donor as well as H-bond-acceptor groups. The highest possible drug loading in the ASDs was smaller for native DIF and the DIF methyl ester derivative (25 wt %) as compared to the methoxy DIF and dimethyl DIF derivatives (35 wt %). Solid-state NMR relaxometry was employed to evaluate the molecular miscibility of the ASD components. In addition, 13C-CPMAS ssNMR spectroscopy was performed on spray-dried ASDs to evaluate the H-bond interactions between the API and polymer. It was observed that the ES100 carboxyl group interacted as an H-bond donor with the C═O carbonyl group of the DIF derivatives, leading to homogeneous mixing for all ASDs. However, it was also shown that competition between intermolecular H-bonds and intramolecular H-bonds, present as stable six-membered rings, is possible. The competition limited the availability of the API acceptor C═O group and explains the lower maximum drug loadings for native DIF and the DIF methyl ester. A comparison was also made between these ES100-based ASDs and PVPVA-based ASDs studied in a previous study. It became clear that for ASDs with a polymer that only carries H-bond-acceptor groups (like PVPVA), the availability of API H-bond-donor groups is crucial for the formation of H-bonds between the drug and the polymer. Contrastingly, for ASDs formed with a polymer that carries both H-bond-donor and -acceptor groups (like ES100), it seems to be the availability of API H-bond-acceptor groups that is crucial for intermolecular H-bond formation and drug-polymer miscibility.

Replacing benzene rings with C(sp[3])-rich bioisosteres to yield compounds with improved drug-like properties is an attractive strategy in medicinal chemistry. While many caged hydrocarbons have been validated as bioisosteres of ortho- and meta-disubstituted benzenes, 3D analogues of 1,2,4-trisubstituted benzenes-the second most prevalent benzenoid pattern in drugs-remain elusive because vector fidelity and enantioselective access are still formidable challenges. Here we report a practical route to (enantiomerically pure) 2-thiabicyclo[3.1.1]heptanes (thia-BCHeps) by cycloadditions of bicyclo[1.1.0]butanes with 1,4-dithiane-2,5-diol. This method produces cycloadducts with two and three exit vectors, which serve as promising bioisosteres for ortho-substituted and 1,2,4-trisubstituted benzenes, respectively. Moreover, the cycloadducts can be transformed into a diverse chemical space, including 1,5-disubstituted thiabicyclo[3.1.1]heptenes. Crystallographic analysis and a comparison of the pharmacokinetic properties, along with an evaluation of the biological activity of diflunisal, salicylanilide and the anticancer drug sonidegib, in relation to their 3D thia-BCHep analogues, demonstrate that the thia-BCHeps obtained can provide new surrogates for 1,2,4-trisubstituted, meta- and ortho-substituted benzene rings in drug discovery programmes.