Kinases have a pivotal role in phosphorylation and kinase dysregulation has been implicated in the occurrence and progression of various diseases. By mid-2024 the Food and Drug Administration had approved 81 small-molecule kinase inhibitors targeting greater than 30 kinases, providing substantial clinical benefits for patients with cancer and other diseases. However, many reported kinase inhibitors exhibit poor target selectivity, limited solubility, and issues with drug resistance. To address these challenges medicinal chemists have used a macrocyclization strategy to optimize these molecules and three macrocyclic kinase inhibitors (lorlatinib [ALK/ROS1], pacritinib [Flt3/JAK2], and repotrectinib [TRK/ROS1]) have received FDA approval. The macrocyclization strategy is now widely used to enhance kinase inhibitory activity, overcome drug resistance caused by point mutations, improve kinase selectivity, and optimize drug-like properties. In this review we provide a comprehensive overview of the utilization of macrocyclization in the optimization of kinase inhibitors by detailing the structural modification process from lead compounds to macrocyclic molecules that has been applied in recent years. In addition, we discuss the limitations of existing macrocyclic kinase inhibitors and propose key considerations for the development of improved macrocyclic molecules. We aim for this review to offer valuable insights for future advances in kinase inhibitor development.

Complex biological mechanisms and unidentified therapeutic targets for amyotrophic lateral sclerosis (ALS) significantly hinder the development of effective treatments. Given these challenges, reliable disease models that accurately replicate ALS phenotypes with relevant biological underpinnings are essential for advancing precision medicine in ALS. Patient-derived induced pluripotent stem cell (iPSC) organoids have emerged as an innovative tool for disease modeling and drug evaluation. Growing evidence highlights the advantages of organoids in replicating ALS phenotypes and supporting drug development. However, challenges remain in utilizing organoids for ALS drug testing and other neurodegenerative diseases. In this review we summarize the current progress in ALS model development, encompassing both in vitro and in vivo non-human models, as well as iPSC-derived human models. Furthermore, within the context of ALS drug screening, we discuss critical considerations for applying organoids to evaluate disease-associated phenotypes and to accurately reflect disease-related symptoms.

The UPLC-QqQ-MS/MS method was established, validated, and used for the simultaneous detection of berberine (BBR), astragaloside IV (AST), and the main metabolites to demonstrate the comparative pharmacokinetics of BBR and AST in normal and T2DM rats. BBR and AST had reduced the internal exposure of each other and their main metabolites in normal rats. However, AST had few significant effects on the pharmacokinetic parameters of BBR and the main metabolites in T2DM rats. Similarly, BBR had no significant effect on the pharmacokinetic parameters of AST but significantly increased the exposure to cycloastragenol (CAG) in vivo. Molecular docking of BBR and AST with the P-glycoprotein (P-gp) was performed, which indicated that both BBR and AST are potential substrates for P-gp. The differences in gut microbiota between normal and T2DM rats were compared by 16S rRNA sequencing. Git microbiota that could produce β-glucosidase and β-xylosidase were highly abundant in T2DM rats. The current study indicated that BBR and AST had reduced oral bioavailability. The gut microbiota was enriched in the intestines of T2DM rats and promoted the hydrolysis of AST to produce CAG, while the drug-drug interaction between AST and BBR was blocked.

Cardiovascular diseases (CVDs) pose a significant threat to human health due to the high mortality and morbidity rates. Traditional drugs often have limited efficacy due to inherent constraints, such as low bioavailability and notable side effects. As a highly regarded therapeutic strategy, nanotechnology offers new perspectives and means for treating CVDs. Nanozyme-based targeted therapeutic drugs specifically address the biological processes in areas affected by CVDs, thereby achieving precise treatment. Compared to traditional drugs, targeted nanozymes offer advantages, such as high efficiency, specificity, controllability, and fewer side effects, showing great efficacy in treating CVDs. This paper first explores the design strategies and mechanisms of nanozyme-based targeted therapy, then introduces its application in key CVDs, such as ischemic stroke, myocardial infarction, and coronary heart disease. Finally, the paper discusses the challenges of introducing targeted nanozymes into clinical applications and the future development prospects in the treatment of CVDs.

Epithelial cell adhesion molecule (EpCAM) is a biomarker for epithelial cell-derived tumors. However, the specific role of EpCAM itself in early-stage hepatocellular carcinoma progression remains unclear, and small molecules targeting EpCAM have not yet been reported. Here, the protein expression profile of EpCAM in tumor-adjacent regions was found to be higher than that in tumor regions, and to be positively associated with the progression of early-stage liver cancer, as well as high frequency of recurrence, cirrhosis, lymph node metastasis, microvascular invasion and cancer stemness, in 68 patients with hepatocellular carcinoma (HCC). In vitro, EpCAM enhanced cancer cell stemness, as reflected by increased abilities of proliferation, self-renewal, migration and invasion, which was counteracted by arenobufagin. Furthermore, arenobufagin inhibited the viability of Hep3B and Huh7 cells with IC₅₀ values of 36.4 nM and 123.4 nM after 72 h of treatment, respectively. Molecular docking data further indicated that arenobufagin binds EpCAM. Moreover, arenobufagin inhibited early progression of HCC through EpCAM in a zebrafish xenograft tumor model mimicking early-stage hepatocellular carcinoma without blood vessels in vivo. This study supports a tumor-promoting role of EpCAM in early-stage hepatocellular carcinoma by facilitating cancer stemness and suggests that arenobufagin might be promising candidate for EpCAM inhibition.

Prostate cancer (PrCa) is the most prevalent urogenital cancer affecting men. PrCa is marked by uncontrolled cellular growth that leads to abnormal enlargement of the prostate gland. The metastatic spread of PrCa is the primary cause of mortality, causing cancer cell dissemination to distant sites, such as bones, the pelvis, and various visceral organs. Key contributors to PrCa progression include genetic mutations, elevated androgen receptor expression, gene amplification, and the rise of androgen receptor splice variants. Although androgen deprivation therapy remains the mainstay for early-stage PrCa treatment, efficacy is temporary because many cases advance to castration-resistant PrCa (CRPC), presenting a significant therapeutic hurdle. This review explores key biomarkers for PrCa and the latest therapeutic strategies for CRPC with a particular focus on the innovative proteolysis-targeting chimera (PROTAC) technology. This approach offers a novel means of degrading target proteins and we discuss how PROTAC holds potential as effective strategies to combat resistance mechanisms in CRPC.

The binding affinity of aptamers to targets has a crucial role in the pharmaceutical and biosensing effects. Despite diverse post-systematic evolution of ligands by exponential enrichment (post-SELEX) modifications explored in aptamer optimization, accurate prediction of high-affinity modification strategies remains challenging. Sclerostin, which antagonizes the Wnt signaling pathway, negatively regulates bone formation. Our screened sclerostin aptamer was previously shown to exert bone anabolic potential. In the current study, an interactive methodology involving the exchange of mutual information between experimental endeavors and machine learning was initially proposed to design a high-affinity post-SELEX modification strategy for aptamers. After four rounds of interactive training (a total of 422 modified aptamer-target affinity datasets with diverse modification types and sites), an antifcial intelligence model with high predictive accuracy with a correlation coefficient of 0.82 between the predicted and actual binding affinities was obtained. Notably, the machine learning-powered modified aptamer selected from this work exhibited 10⁵-fold higher affinity (picomole level K_D value) and a 3.2-folds greater Wnt-signal re-activation effect compared to naturally unmodified aptamers. This approach harnessed the power of machine learning to predict the most promising high-affinity modification strategy for aptamers.

Single-cell RNA sequencing (scRNA-seq) data from published datasets were obtained to investigate the expression and dysregulation of RNA-binding proteins (RBPs), which are critical for alternative mRNA splicing and translational control in rheumatoid arthritis (RA). How RBP regulation differs between RA and osteoarthritis (OA) was examined using RBP for single-cell sub-clustering. Quantitative polymerase chain reactions (PCRs) were performed to confirm differentially expressed RBPs in RA fibroblast-like synoviocytes (FLSs) and OA-FLSs, as well as in mice with collagen-induced arthritis (CIA) and control mice. Additionally, bulk RNA-seq data were collected and RBP-alternative splicing event (ASE) co-expression analyses were performed to reveal the potential regulatory role of RA-related RBPs on ASEs. Significant variations in relative proportions of cell subtypes were demonstrations between RA and OA with downregulated RBPs outnumbering upregulated RBPs in each cell type and showing high specificity for particular subsets. One hundred five upregulated and 133 downregulated RBPs were identified in fibroblasts. Y-Box binding protein 3 (YBX3) and splicing factor 3b subunit 6 (SF3B6) were confirmed to be upregulated in RA-FLS and CIA mice, while eukaryotic translation initiation factor 4A1 (EIF4A1) and U2 small nuclear RNA auxiliary factor 1 (U2AF1) were downregulated in RA-FLS. The RA group displayed stronger cell type interactions compared to the OA group with enhanced signaling pathways, such as fibronectin 1-cluster of differentiation 44 (FN1-CD44) and C-X-C motif chemokine ligand 12-C-X-C motif chemokine receptor 4 (CXCL12-CXCR4). Furthermore, three upregulated genes (spectrin repeat containing nuclear envelope protein 2 [SYNE2], S100 calcium binding protein A9 [S100A9], and interferon induced protein with tetratricopeptide repeats 3 [IFIT3]) and four downregulated genes (ribonuclease 1 [RNASE1], granulin [GRN], FN1, and sorbin and SH3 domain containing 2 [SORBS2]) were co-expressed in RA-associated RBPs and ASEs. These findings suggest that dysregulation of RBPs may contribute to the development of RA and provide potential targets for therapeutic interventions.

Estrogens have been reported to cause dysfunction in biliary transport systems, thereby inducing cholestasis. Multidrug resistance-associated protein 2 (MRP2) is a transporter responsible for independent bile flow. Emerging evidence indicates that PDZ domain containing 1 (PDZK1) regulates localization of MRP2; however, PDZK1’s role and regulatory machinery in MRP2-mediated estrogen-induced cholestasis (EIC) remain unclear. Herein, in a mouse model of EIC, we observed downregulated PDZK1 expression in the liver and enhanced intracellular domain MRP2 internalization. Notably, expression of miR-128-3p, a potential biomarker of estrogen-related cholestasis discovered by our group, was significantly elevated. We demonstrated that miR-128-3p targeted the 3’-untranslated region of PDZK1 in EIC and consequently promoted MRP2 internalization. Accordingly, miR-128-3p suppression upregulated PDZK1, thereby suppressing MRP2 internalization and significantly attenuating cholestatic liver disease. Furthermore, we observed MRP2 internalization and PDZK1 downregulation, as well as excessive miR-128-3p, in clinical samples from patients with cholestatic liver injury. Overall, our findings illustrate that miR-128-3p inhibits PDZK1 expression, thereby inhibiting the membrane localization of MRP2 in EIC. Enhancing or restoring PDZK1 expression might therefore have therapeutic potential for cholestatic liver injury.

Isosteviol and its derivatives, diterpene compounds with a wide range of biological activities, play important antiviral, antibacterial, antioxidant, antitumor, analgesic, anti-inflammatory, antipyretic, antifungal, and anticardiovascular disease roles. Because of its unique diterpene skeleton, isosteviol is frequently used as an active framework in drug synthesis, thus helping medicinal chemists design highly selective, potentially active, and multifunctional isosteviol analogs for the treatment of various diseases. In recent years, rapid developments in the design and synthesis of isosteviol derivatives have proven effective and important in the field of medicinal chemistry research. This article briefly reviews the novel derivatives obtained through structural modification of isosteviol, including their pharmacological activities, to provide a reference for the development of new synthetic strategies and the construction of new isosteviol derivatives, and to guide preclinical studies and development of new drugs with greater pharmacological efficacy.