Background/Objectives: Astragalus root is a classical qi-tonifying traditional Chinese medicine that has demonstrated potential therapeutic efficacy in type 2 diabetes mellitus (T2DM) and non-alcoholic fatty liver disease (NAFLD). However, the precise mechanisms underlying its effects on the comorbidity of these two disorders remain unclear. This study investigated the molecular mechanisms by which Astragalus root ameliorated T2DM-NAFLD comorbidity. Methods: Network pharmacology, molecular docking, molecular dynamics simulation, and in vitro experiments were employed to elucidate the potential roles and mechanisms of Astragalus root in the management of T2DM-NAFLD comorbidity. Results: A total of 25 bioactive constituents and 152 corresponding targets associated with Astragalus root were identified. PPI network analysis revealed the top ten core candidate targets, among which six possessed suitable crystal structures for molecular docking, including interleukin-6 (IL-6), threonine-protein kinase 1(AKT1), transcription factor AP-1(JUN), tumor necrosis factor (TNF), cysteine-dependent aspartate-specific protease 3 (CASP3), and estrogen Receptor 1(ESR1). Kyoto encyclopedia of genes and genomes (KEGG) analysis further identified the phosphatidylinositol 3-kinase (PI3K)-AKT as the most significantly enriched pathway. Molecular docking validated the potential binding modes of formononetin to the six core targets, a finding that was further confirmed by molecular dynamics simulations, which proved the stability of the resulting complexes. In vitro experiments demonstrated that formononetin obviously decreased lipid droplet accumulation, downregulated total cholesterol (TC) and triglyceride (TG) levels, suppressed the expression of TNF-α, IL-6, and interleukin-1β (IL-1β), decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels, and enhanced glutathione (GSH) content and superoxide dismutase (SOD) activity. These therapeutic effects were achieved through inhibition of protein expression within the PI3K/AKT/mechanistic target of rapamycin (mTOR) signaling pathway. Conclusions: This study determined the potential therapeutic targets and underlying mechanisms of formononetin derived from Astragalus root in the T2DM-NAFLD management, thereby providing a scientific basis for its clinical application.

The optimization of membrane permeability is a pivotal approach for mitigating late-stage failures in peptide drug development. By leveraging linker chemical diversity, stapled peptides utilize linker engineering to precisely modulate key physicochemical parameters—such as lipophilicity and conformational constraints—to overcome the desolvation energy penalty. This review systematically evaluates linker-based strategies for enhancing the permeability of stapled peptides, categorized into two primary dimensions: (1) high-throughput screening (HTS) compatibility, focusing on the integration of functionalized linkers into mRNA display, phage display, and DNA-encoded libraries (DELs) to identify lead scaffolds with inherent permeability potential during early discovery; and (2) post-screening structural refinement, covering rational design strategies including intramolecular hydrogen-bond (IMHB) shielding, “chameleonic” adaptations, and stimuli-responsive reversible stapling. Furthermore, we analyze the paradigm shift in assessment methodologies from qualitative imaging to quantitative cytosolic delivery assays, which have deepened our understanding of mechanisms such as the charge/lipophilicity threshold balance and metabolism-driven trapping. Overall, linker engineering provides a robust technical roadmap for developing the next generation of cell-permeable stapled peptide therapeutics.

Background/Objectives: Astragalus root is a classical qi-tonifying traditional Chinese medicine that has demonstrated potential therapeutic efficacy in T2DM and NAFLD. However, the precise mechanisms underlying its effects on the comorbidity of these two disorders remain unclear. This study investigated the molecular mechanisms by which astragalus root ameliorated T2DM-NAFLD comorbidity. Methods: Network pharmacology, molecular docking, molecular dynamics simulation, and in vitro experiments were employed to elucidate the potential roles and mechanisms of astragalus root in the management of T2DM-NAFLD comorbidity. Results: A total of 25 bioactive constituents and 152 corresponding targets associated with astragalus root were identified. PPI network analysis revealed the top ten core candidate targets, among which six possessed suitable crystal structures for molecular docking, including IL-6, AKT1, JUN, TNF, CASP3, and ESR1. KEGG analysis further identified the PI3K-AKT as the most significantly en-riched pathway. Molecular docking of the principal bioactive constituent formononetin from astragalus root with the six core targets was conducted using AutoDock4 software. Molecular dynamics simulations verified the stability of the interactions between for-mononetin and each of the six core target proteins. In vitro experiments demonstrated that formononetin obviously decreased lipid droplet accumulation, downregulated TC and TG levels, suppressed the expression of TNF-α, IL-6, and IL-1β, decreased ROS and MDA levels, and enhanced GSH content and SOD activity. These therapeutical effects were achieved through inhibition of protein expression within the PI3K/AKT/mTOR signaling pathway. Conclusions: This study determined the potential therapeutic targets and underlying mechanisms of formononetin derived from astragalus root in the T2DM-NAFLD management, thereby providing a scientific basis for its clinical application.

The optimization of membrane permeability is a decisive strategy for mitigating late-stage failures in peptide drug development. By leveraging linker chemical diver-sity, stapled peptides utilize linker engineering to precisely modulate key physico-chemical parameters—such as lipophilicity and conformational constraints—to over-come the desolvation energy penalty. This review systematically evaluates link-er-based strategies for enhancing the permeability of stapled peptides, categorized into two primary dimensions: (1) High-throughput screening (HTS) compatibility, focusing on the integration of functionalized linkers into mRNA display, phage display, and DNA-encoded libraries (DELs) to identify lead scaffolds with inherent permeability potential during early discovery ; and (2) Post-screening structural refinement, cover-ing rational design strategies including intramolecular hydrogen bond (IMHB) shield-ing, "chameleonic" adaptations, and stimuli-responsive reversible stapling . Further-more, we analyze the paradigm shift in assessment methodologies from qualitative imaging to quantitative cytosolic delivery assays, which have deepened our under-standing of mechanisms such as the charge/lipophilicity threshold balance and meta-bolic-driven trapping. Overall, linker engineering provides a robust technical roadmap for developing the next generation of cell-permeable stapled peptide therapeutics.

Glycyrrhizic acid class compounds are used empirically for checkpoint inhibitor-induced liver injury (ChILI), but their effects on antitumour efficacy remain uncertain. This narrative review examines isomer-specific pharmacology, clinical evidence and the candidate mechanisms proposed for this class, of which the high-mobility group box 1 (HMGB1) axis is the most developed. In a propensity-score-matched cohort of 229 patients, magnesium isoglycyrrhizinate (MgIG) plus corticosteroids was associated with higher day-10 alanine aminotransferase normalization than corticosteroids alone (30.7% versus 17.4%). A single-arm study of 67 patients reported study-defined liver injury in 9.0% during prophylaxis. Neither study reported antitumour efficacy or established causal clinical benefit. Modulating extracellular HMGB1 might reduce organ inflammation while altering tumor immunosuppression. However, neither direct HMGB1 engagement by MgIG nor a causal contribution of HMGB1 to ChILI was established in the identified literature. HMGB1 can also support antitumour immune priming; its net effect depends on redox state, tissue and timing. The corresponding 18α aglycone shows no measurable inhibition of 11β-hydroxysteroid dehydrogenase type 2 up to 20 µM in vitro, which does not establish clinical safety. Controlled trials should assess liver injury, corticosteroid exposure, antitumour outcomes and electrolyte safety together; routine prophylaxis is not supported.

Background/Objectives: Astragalus root, a traditional Chinese herbal remedy, has shown potential benefits against diabetic nephropathy (DN). However, the mechanisms driving its effects remain poorly understood. This study explored the molecular pathways through which Astragalus root improves DN. Methods: To identify possible targets and mechanisms of Astragalus root in DN treatment, we applied network pharmacology, molecular docking, molecular dynamics simulation, and in vitro assays. Results: Network pharmacology screening uncovered 46 overlapping targets between Astragalus root and DN. Protein-protein interaction (PPI) network analysis identified five core candidate targets: CASP3, VEGFA, CTNNB1, MYC, and PRKCB. KEGG pathway analysis indicated that the AGE-RAGE signaling pathway was the most significantly enriched. Molecular docking revealed that quercetin, β-carotene, daidzein, capsaicin, and kaempferol—major bioactive components of Astragalus root—bound strongly to each of the five core targets. Molecular dynamics simulations further confirmed the conformational stability of kaempferol when complexed with these target proteins. In vitro experiments showed that kaempferol markedly reduced protein levels of α-SMA, Col I, and Col IV; lowered secretion of TNF-α, IL-6, and IL-1β; and decreased ROS and MDA content. Additionally, kaempferol's therapeutic effects were mediated through suppression of the AGE-RAGE-PKC-TGF-β signaling axis. Conclusions: This work identified kaempferol, a bioactive ingredient of Astragalus root, as a potential therapeutic agent against DN, along with its target pathways. These findings provide a scientific foundation for its clinical translation.