Microneedles (MNs), as innovative biomedical devices, are undergoing a transformation from basic transdermal devices to intelligent and multifunctional systems. The advancement of intelligent MNs enhances tissue penetration and adhesion, triggers timely, on-demand and precise spatiotemporal controlled release, achieves sequential drug delivery and in situ monitoring, and enables close-loop theranostics via the integration of flexible electronics, wireless communication, and artificial intelligence. This review not only systematically highlights these cutting-edge breakthroughs, focusing on five major innovative advancements of conceptual design, including mimicking strategies, stimuli-responsive systems, innovative structural designs, living payload, and wearable integration, but also delineates the principal obstacles obstructing the advancement of intelligent MNs. We also emphasize that MN technology is fueling the evolution of next-generation medical paradigms based on its personalized, minimally invasive, and intelligent features through interdisciplinary integration, which is crucial for reshaping the future landscape of disease diagnosis and treatment.

The limited aqueous solubility of active pharmaceutical ingredients (APIs) remains a major challenge in drug development, severely compromising clinical performance. Crystal engineering has emerged as a powerful and versatile approach to address this issue by rationally designing API crystal structures through precise control of intermolecular interactions, thereby enhancing solubility, dissolution rates, and ultimately bioavailability. This review systematically summarizes recent advances in crystal engineering strategies for poorly water-soluble drugs, including polymorphs, cocrystals, solvates/hydrates, nanocrystals, organic framework solids, solid solutions, liquid crystals, amorphous solids, and salts. Additionally, key challenges in translational applications are discussed, including structure-property relationship, AI-driven computational modeling, in vitro-in vivo correlation establishment, and advanced crystallization techniques. The review aims to provide strategic insights of crystal engineering for overcoming solubility barriers in next-generation drug formulations.

Proteins have emerged as highly promising biomaterials for the design of drug-loaded nanocarriers due to their biocompatibility, biodegradability and reduced immunogenicity. Among them, albumin stands out as the most widely used due to its unique physicochemical and biological properties, high affinity to important cell surface receptors, structural stability, long circulation time and intrinsic binding capacities. This review provides an overview of the advantages and limitations of the main proteins that have been proposed as biomaterials for nanoparticle fabrication, with a specific focus on albumin-based systems. It explores the physicochemical characteristics of these nanosystems, receptor binding affinity and functionalization strategies for both passive and active tumor targeting. The main synthesis methods and functionalization strategies are discussed, highlighting their relevance in cancer therapy. Their clinical relevance is stressed by the US Food and Drug Administration (FDA)-approved formulations and the additional albumin-bound drugs in ongoing trials. Despite promising preclinical data and numerous active targeting approaches reported, clinical translation remains limited. This review provides the necessary information to develop improved strategies and cover the gap between preclinical research and clinical application and outlines future perspectives for enhancing the therapeutic efficacy and specificity of albumin-based drug delivery nanosystems in oncology.

Outer membrane vesicles (OMVs) are nanoscale lipid-bilayer vesicles naturally released by Gram-negative bacteria. By packaging membrane proteins, lipopolysaccharide-derived pathogen-associated molecular patterns, nucleic acids, and metabolites, OMVs integrate intrinsic bioactivity with cargo capacity and have emerged as a versatile platform for therapeutic delivery and immune modulation. In this review, we summarize current understanding of OMV biogenesis, composition, and physicochemical features that shape biodistribution and immunogenicity. We then discuss engineering strategies ranging from genetic rewiring of parental strains and detoxification of lipid A to surface functionalization, hybrid membrane assembly, and stimuli-responsive formulations that enable controllable targeting, loading, and release. Recent progress is highlighted in anti-tumor therapy, nanovaccines for infectious diseases and cancer, and nucleic acid delivery for gene regulation. Finally, we analyze key barriers to clinical translation, including endotoxin-associated safety, batch-to-batch heterogeneity, scalable manufacturing, and standardization of quality attributes. Addressing these challenges through rational design and robust production pipelines will be essential to advance OMV-based therapeutics toward safe, effective, and manufacturable clinical products.

The remarkable clinical success of mRNA-lipid nanoparticle (mRNA-LNP) vaccines, particularly evidenced by their pandemic-era impact and subsequent 2023 Nobel Prize recognition, has intensified global efforts to expand their application toward therapeutic biologics. Despite showing strong preventive effects, there are still fundamental knowledge gaps in comprehensively defining the hierarchical biological trajectory of mRNA-LNP formulations, which spans from initial systemic exposure, tissue-specific biodistribution, and intracellular delivery to ultimate protein expression dynamics. These unresolved mechanistic questions pose significant constraints on the rational design of next-generation mRNA-LNP therapeutics, which face increased demands for precision dosing, therapeutic efficacy optimization, and enhanced safety profiles. This review discusses the in vivo fate of mRNA-LNP, focusing specifically on blood/tissue/cellular distribution and kinetics/exposure profiles of both intact mRNA-LNP and dissociated components post-administration. It discusses key factors influencing the distribution of expressed protein and summarizes the corresponding research toolkits along with their advantages and disadvantages. Additionally, it compiles strategies to enhance delivery efficacy, tissue tropism, and safety profiles of mRNA-LNP. We anticipate a better understanding of the biological processes and dynamic pattern of mRNA-LNP, which will accelerate the development of next-generation mRNA biologics with predictable safety and delivery efficiency.

Cancer metastasis is a critical indicator of cancer progression and serves as a major cause of cancer-related deaths. Cuproptosis is a novel form of regulated cell death proposed in 2022. Unlike ferroptosis and other known regulated cell deaths (RCDs), cuproptosis has a unique regulatory pathway, and its major biochemical features include copper overload, lipoylated tricarboxylic acid cycle protein aggregation, and the loss of iron-sulfur cluster protein. Cuproptosis-based nanomedicine provides novel therapeutic insights for metastatic cancer treatment. The close link between cuproptosis and cancer therapy has been explored, and several therapeutic strategies have been developed, including copper ionophores and drug delivery systems. Cuproptosis-based nanotherapeutic strategies may enable controlled and selective drug release, and through the multifaceted actions of copper metallocompounds, achieve multimodal theranostic modalities or organically synergize with other regulated RCD pathways. This integration enhances antitumor effects and biosafety, overcomes tumor resistance, and improves the tumor microenvironment. In this review, we systematically delineate the mechanisms of cuproptosis and its current therapeutic implications in metastatic malignancies. We further critically analyze these synergistic therapeutic approaches, prospect emerging applications of cuproptosis-based nanomedicine in metastatic oncology, and highlight their untapped therapeutic potential. Ultimately, we anticipate this exploration will inform innovative clinical management strategies for cancer patients with metastasis.

The introduction of environment-responsive probes has greatly improved the accuracy of fluorescence bioimaging in evaluating drug nanocarriers. This review highlights the key roles of Förster resonance energy transfer, aggregation-induced emission, and aggregation-caused quenching in advancing nanomedicine. These technologies have enhanced our understanding of nanocarrier pharmacokinetics, biodistribution, and intracellular behavior, providing valuable insights for optimizing drug delivery systems. Their integration into imaging platforms has enabled precise monitoring of nanocarriers in complex biological environments. This review outlines detailed progress in the use of environment-responsive probes, emphasizing their importance in improving the design and effectiveness of nanomedicines. Looking forward, advances in probe engineering and multimodal imaging, combined with computational tools, are expected to drive the development of more targeted, efficient, and personalized therapeutic strategies.

Oral dosage is the most commonly used and preferred method of drug administration due to its several advantages, including non-invasiveness, patient adherence, and ease of use. However, oral bioavailability can be influenced by several factors, such as drug solubility and mucosal permeability in the gastrointestinal tract, sometimes leading to poor and/or inconsistent absorption. In particular, low aqueous solubility presents a significant challenge in achieving adequate oral bioavailability and therapeutic effectiveness for many pharmaceutical compounds. Attempts to overcome these limitations have focused on deeper understanding of the physicochemical, biochemical, and biological barriers that limit overall drug bioavailability. To ensure better and stable pharmacokinetic behavior of orally administered drugs, various formulation strategies have been developed to enhance solubility, dissolution rate, membrane permeability, and overall oral bioavailability. This review article explores recent advancements in formulation techniques aimed at improving the biopharmaceutical properties of orally administered drugs. The challenges and development aspects of oral dosage forms are also addressed.

Lyotropic liquid crystals (LLCs), formed through the self-assembly of amphiphilic molecules in polar solvents, offer thermodynamic stability and tunable mesophases (lamellar, hexagonal, and cubic), making them a versatile platform for biomedical applications. Their structural adaptability enables enhanced drug stability, improved bioavailability, and controlled release, which are advantageous for various therapeutic strategies. LLC-based systems have shown significant promise in drug delivery, long-acting therapies, anti-infective treatments, and wound healing. Their diverse formulation options, including gels, nanoparticles, and in situ forming precursors, support multiple routes of administration, such as oral, intravenous, dermal, ocular, and intranasal. This review summarizes recent advances in the design and functionalization of LLC systems, with a focus on their ability to overcome physiological barriers, enhance therapeutic efficacy, enable targeted delivery, and support prolonged treatment regimens. Challenges in clinical translation and future research directions are also discussed to facilitate the transition from bench to bedside.

The global pharmaceutical drug delivery market is forecasted to grow to USD 2546.0 billion by 2029. The expanding pharmaceutical market urgently needs a more efficient drug research and development paradigm. Artificial intelligence (AI) is revolutionizing drug delivery by offering alternatives to traditional trial-and-error experimental approaches. This review systematically traces the technological evolution from early simple models to current advanced AI algorithms in various applications, ranging from formulation optimization to the prediction of critical formulation parameters and de novo material design. To enhance the reliability of AI applications in drug delivery, we present comprehensive guidelines and “Rule of Five” (Ro5) principles to systematically direct researchers in utilizing AI in formulation development. This “Ro5” includes the following criteria: a formulation dataset containing at least 500 entries, coverage of a minimum of 10 drugs and all significant excipients, appropriate molecular representations for both drugs and excipients, inclusion of all critical process parameters, and utilization of suitable algorithms and model interpretability. The review concludes with insights into emerging trends and future directions, including the utilization of large language models, multidisciplinary collaboration opportunities, talent development, and culture transformation, aimed at facilitating a paradigm shift toward AI-driven drug formulation development.

The treatment of interstitial lung diseases (ILDs) was important to reduce the inflammation or fibrosis within the interstitial space. In recent years, a variety of undruggable ILDs targets were emerged for anti-inflammation and anti-fibrosis therapy. The development of RNA delivery system provided the potential for undruggable targets to the lung via inhalation. However, the RNA delivery systems still faced the challenges, including the protection of the stability of RNA platform, increasing the effective delivery to the targeted cells, and selective escaping of RNA molecules into the cytoplasm. In this review, we first summarized the physiological and biological barriers of RNA inhaled platform. Subsequently, the progress of inhaled RNA delivery system and their therapeutic efficiency have been systematically addressed for the application of ILDs. Finally, in the design of an inhaled RNA delivery system, key factors needed to consider and perspectives are discussed.

Advancements in drug discovery, such as artificial intelligence, computational technology, and combinatorial chemistry, have led to numerous new promising drug candidates. However, most still fail to reach the market due to poor physicochemical properties, resulting in off-target or toxic effects. As a potential solution, non-invasive and targeted drug delivery at mucosal surfaces can deliver drugs directly to therapeutic sites to avoid off-target effects, decrease required drug doses, bypass hepatic first-pass metabolism, and circumvent uncontrolled drug release. Liquid-based drug delivery systems have advanced tremendously over recent years, with novel dosage forms such as ionic liquids, liquid crystals, stimuli-responsive phase transforming liquid systems, nanoemulsions, double-emulsions, self-emulsifying delivery systems, and eutectic systems being developed. These systems hold vast promise for transmucosal drug delivery as liquids generally provide superior spreadability compared to solid dosage forms, and some liquid systems provide prolonged mucosal retention times compared to conventional solutions. However, liquid dosage forms present regulatory challenges such as preservation, sterility, and stability requirements. This review discusses novel liquid-based formulation approaches to cross the ocular-, nasal-, oromucosal-, and vaginal mucosal barriers emphasizing advanced liquid drug delivery systems and the progress made toward clinical translation as a platform for future transmucosal liquid-based dosage form development.

Atherosclerosis, the leading cause of cardiovascular diseases, has become increasingly prevalent worldwide, driving the need for innovative therapeutic strategies to improve clinical outcomes. Lipid-based nanosystems have emerged as promising drug delivery platforms due to their biocompatibility, versatility, and proven clinical track records. Among them, traditional systems such as liposomes have been extensively applied in atherosclerosis therapy, primarily for their well-established safety profile and their capacity to deliver both hydrophilic and hydrophobic agents. However, emerging research has expanded the scope of lipid-based nanosystems to include lipid nanoparticles, lipoprotein-based nanosystems, cell membrane-coated nanosystems, and so on. These systems have demonstrated great promise in addressing the complex and heterogeneous nature of the disease. This review aims to provide a comprehensive overview of lipid-based nanosystems, from traditional formulations to cutting-edge innovations, and their evolving applications in the treatment of atherosclerosis. We also discuss the challenges associated with the clinical translation of these systems, as well as future prospects for developing more effective and personalized therapeutic strategies. In conclusion, lipid-based nanosystems provide a promising option for atherosclerosis treatment, potentially driving the progress of novel therapeutic strategies.

This review explores the molecular and cellular pathways of soft tissue wound healing and the potential therapeutic use of the non-psychotropic cannabinoid cannabidiol (CBD), integrating findings from in vitro and in vivo preclinical studies as well as completed and ongoing clinical trials. It provides a comprehensive summary of the next steps in new CBD-based product development by analyzing current trends in dosage optimization, treatment guidance, delivery systems, ranging from liposomes, microemulsions to hydrogels. Additionally, the review examines clinical trials related to CBD formulations, delivery routes, and participant outcomes, offering a deeper understanding of the mechanisms guiding the activity beyond binding to cannabinoid 1 (CB1) and CB2 receptors. Furthermore, it highlights challenges and future perspectives in CBD formulation studies, presenting both currently studied approaches and emerging possibilities for innovation. Therapeutic potential of CBD has proved itself in the recent years and only regulatory issues and clarity in treatment and delivery routes will limit its widespread use in soft tissue healing.

Cancer immunotherapy is an innovative treatment approach that leverages the immune system's ability to identify and attack tumor cells. Its goal is to initiate or re-establish the tumor-immune cycle. However, challenges such as limited response rates and adverse immune responses have hindered the further application and advancement of this therapy. Recent progress in nanomedicine, particularly in self-assembled nanomaterials, has attracted significant attention due to their excellent physical and chemical properties. Self-assembled nanoplatforms can be designed to selectively deliver immunoadjuvants, therapeutic drugs, photosensitizers, and sonosensitizers, overcoming the limitations of traditional monotherapies. By utilizing these self-assembled nanoplatforms to synergistically combine cancer immunotherapy with photodynamic therapy (PDT), photothermal therapy (PTT), radiotherapy (RT), and sonodynamic therapy (SDT), it becomes possible to amplify and enhance the immune responses elicited by these localized treatments, thus offering new strategies for cancer therapy. In this review, we discussed various immunotherapy platforms based on the self-assembly of nucleic acids, peptides and proteins, metals, and supramolecules. We also highlight the significant research advancements over the past three years in the use of self-assembled nanomaterials for combination therapies centered on immunotherapy, aiming to provide valuable insights and references for ongoing tumor therapy research.

Autoimmune diseases are characterized by an aberrant immune response directed against the body's own components, leading to immune dysfunction and loss of tolerance. This ultimately results in tissue damage and organ impairment. Despite the availability of a range of pharmacological agents for the treatment of autoimmune diseases, numerous challenges still remain. These include reduced bioavailability, first-pass metabolism, severe adverse effects, and low patient compliance associated with oral or injectable routes of administration. Microneedles (MNs) based delivery systems offer a promising alternative for transdermal drug administration, as they can circumvent the aforementioned limitations via the generation of microchannels in the skin in a minimally invasive manner with low pain. This review examines the applications and recent advances in MN systems for treating autoimmune diseases, focusing on six key areas: an overview of autoimmune diseases, current treatment modalities, an introduction to MNs, advantages of MNs for autoimmune disease treatment, advancements in MN-mediated therapies for diverse autoimmune diseases, and relevant MN technologies currently undergoing clinical trials. Additionally, it considers the prospective future advancements of MN systems in the management of autoimmune diseases.

Despite the different degrees of blood-brain barrier (BBB) damage in diverse brain diseases, it remains a formidable barrier that restricts most drugs from penetrating the brain. A comprehensive understanding and elucidation of the disease-specific changes of BBB in various brain pathologies are essential for directing the customized brain-targeted drug delivery systems, potentially improving cerebral delivery efficiency and therapeutic efficacy. Hence, this review compared anatomical and physiological changes of BBB under healthy and pathological states and discussed the effects of these changes on cerebral delivery efficiency. Thereafter, a particular emphasis was placed on the pathology-directed drug delivery strategies tailored to different brain diseases, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, stroke, and brain tumors. By combining insights from cutting-edge studies and emerging technologies, we proposed forward-looking suggestions on future directions to brain-targeted drug delivery, thereby improving the therapeutic efficacy and accelerating the translation from preclinical attempts into clinical practice.

The limited repertoire of experimentally validated RNA-targeting nucleases has constrained both mechanistic studies and the efficient discovery of novel enzymes for RNA biotechnology. This challenge is particularly pronounced for prokaryotic Argonaute (Ago) proteins, where the scarcity of confirmed RNA-targeting members and a lack of clarity regarding RNA specificity determinants hinder systematic exploration. Although machine learning offers a potential solution, its application is often impeded by the scarcity of labeled training data in this field. To address these limitations, we developed the self-iterative hierarchical ensemble model (SIM), which integrates hierarchical ensemble learning with a self-training strategy. This approach bypasses the dependency on large-scale experimental datasets, allowing SIM to iteratively expand its predictive capability from minimal initial labeled data. When applied to prokaryotic Agos, SIM identified six high-confidence RNA-targeting candidates, five of which were experimentally validated (83% success rate). Notably, SIM identified three uncharacterized Agos harboring a novel N-terminal domain, defining a previously unrecognized subclass. Biochemical and in vivo validations of Haloferax profundi Ago (HpAgo) confirmed its RNA cleavage activity and a distinctive RNA modification-sensing capability. We leveraged this latter finding to develop a rapid, cost-effective method for quantifying modified RNAs. Our study not only expands the repertoire of RNA-targeting tools but also establishes SIM as a generalizable framework for protein function prediction under data-scarce conditions. This work has broad implications for both RNA biotechnology and the application of machine learning in data-limited fields.

Gene essentiality (synonymous with dependency) mapping reveals therapeutic vulnerabilities for intractable oncogenes, yet integrated platforms bridging CRISPR functional genomics with drug discovery remain limited. To address this gap, we developed GEMap, A Gene Essentiality-Guided Platform for Drug Discovery, by combining genome-wide CRISPR-Cas9 essentiality profiles (1912 screens across 1135 cancer cell lines) with multi-omics annotations, drug profiles and drug targets information (20,000+ compounds). GEMap can be used to (1) explore multimodal gene data (Dependency/Expression/CNV/Mutation) across cell lines and tissues; (2) prioritize context-specific therapeutic targets by quantifying differential genetic dependencies in molecularly stratified cancers, such as KRAS mutant-cancers; and (3) discover tailored treatments and drug candidates targeting particular gene using large-scale drug perturbation data and drug physical targets. To the best of our knowledge, GEMap represents the first platform bridging CRISPR-derived genetic dependencies with pharmacological responses and biological networks and establishes an open-access paradigm for accelerating precision oncology against undruggable targets. GEMap is available at https://web.biotcm.net/GEMap/.

The suppressive microenvironment of AML limits anti-PD-1 efficacy, making pyroptosis induction a key strategy for its remodeling. Isoalantolactone (IAL), a naturally occurring small molecule, has been identified herein to inhibit cytosolic thioredoxin reductase1 (TXNRD1) and trigger pyroptosis in AML cells, thereby enhancing the efficacy of anti-PD-1 antibody therapy. Mechanistically, the α,β-unsaturated carbonyl group of IAL covalently bound to the selenocysteine residue at the position 498 (Sec⁴⁹⁸) of TXNRD1 through a Michael addition reaction. Clinical sample analysis revealed that TXNRD1 is overexpressed in AML, which correlates with poor prognosis. Additionally, we found that the TXNRD inhibitor auranofin has demonstrated good efficacy against AML. The inhibition of TXNRD1 activates the transcription factor peroxisome proliferator-activated receptor gamma (PPARγ), which, in turn, upregulates the transcription of caspase-3, subsequently increasing the cleavage of gasdermin E to induce pyroptosis via the non-classical pathway in AML cells. Additionally, enhanced caspase-3 activity promotes poly ADP-ribose polymerase 1 (PARP-1) cleavage and upregulates PD-L1 expression, thereby increasing sensitivity to anti-PD-1 monoclonal antibodies. Overall, the current study highlights a promising approach for augmenting therapy using anti-PD-1 monoclonal antibodies in AML by targeting TXNRD1 to induce pyroptosis and ameliorate the immunosuppressive microenvironment.

Attenuated Salmonella VNP20009 (VNP) shows promising anti-cancer therapeutic potential. Limited understanding of its anti-tumor mechanism has hindered broader clinical application. Recent studies have reported cellular senescence is involved in tumor progression; however, its critical role in VNP therapy remains elusive. Our study revealed that VNP exerts anti-tumor growth and anti-angiogenesis effects by inducing cellular senescence in tumor cells and vascular endothelial cells. While VNP-induced senescence inhibits tumor growth, it concurrently promotes neutrophil extracellular traps (NETs), which paradoxically enhance tumor progression. To address this challenge, we engineered VNP-SNase, a novel variant capable of releasing the DNA-degrading enzyme Staphylococcus aureus nuclease directly within tumors. VNP-SNase significantly inhibited NETs formation across multiple tumor types, effectively promoted anti-tumor immunity, and exhibited improved tumor suppression effects with enhanced biosafety. Our findings elucidate the critical role of cellular senescence in VNP therapy and propose targeting NETs as a strategic approach to enhance the efficacy of VNP-based cancer treatments.

Prenatal herbicide exposure is increasingly linked to neurodevelopmental disorders, yet effective pharmacological interventions remain lacking due to unclear pathogenic mechanisms. Here, we demonstrate that prenatal exposure to glufosinate ammonium (GLA), a widely used herbicide, triggers autism-like behaviors, including social deficits and repetitive grooming, in offspring mice. Whole-brain c-Fos mapping, in vivo calcium imaging, and patch-clamp recordings identified hypoactive pyramidal neurons in the anterior cingulate cortex (ACC) as the neural substrate of these behavioral deficits in prenatally GLA-exposed offspring mice. Mechanistically, transcriptomic and multi-omics analyses revealed that astrocyte activation in the ACC drove Kir4.1 potassium channel upregulation, which suppressed CaMKIIα⁺ neuronal excitability via impaired astrocyte-neuron communication. Pharmacological inhibition of astroglial Kir4.1 not only restored neuronal activity but also rescued social deficits in GLA-exposed offspring, underscoring Kir4.1’s pivotal role in ACC dysfunction. Our study uncovers a novel astrocyte-neuron axis underlying herbicide-induced neurodevelopmental impairments and identifies Kir4.1 as a therapeutic target for environmental factor-associated autism.

Prednisone is widely used to treat pregnancy-complicated autoimmune diseases, but the impact of prenatal prednisone exposure (PPE) on offspring neurodevelopment is unclear. Clinical follow-up using the Ages and Stages Questionnaires (ASQ:SE-2 and ASQ-3) revealed a strong association between PPE and neurodevelopmental abnormalities. In a rat model, PPE significantly inhibited the proliferation and differentiation of cortical radial glial cells (RGCs), impairing cortical neurogenesis and leading to anxiety- and depression-like behaviors. Mechanistically, PPE upregulated fat mass and obesity-associated protein (FTO) nuclear recruitment in RGCs, depleting N⁶-methyladenosine (m⁶A) modifications and stabilizing Crebbp/Ep300 mRNA, which activated genes related to cell cycle arrest and neuronal differentiation via histone 3 at lysine 27 acetylation (H3K27ac) modifications. The key role of FTO in PPE-induced neurodevelopmental impairments was confirmed using Ep300 shRNA, Fto overexpression lentivirus, and a conditional Fto knockout mouse model (Ftofl/fl; Nestin-Cre). Notably, early postnatal neuregulin-1 supplementation promoted RGCs proliferation and alleviated behavioral abnormalities.

Pepducins are synthetic membrane-tethered lipopeptides designed to allosterically modulate G protein-coupled receptor (GPCR) signaling. Here, we characterize a series of pepducins targeting the neurotensin receptor type 1 (NTSR1), revealing their complex and multifaceted modulation properties. Using BRET-based biosensors, we show that PP-001, a pepducin derived from NTSR1’s first intracellular loop, preferentially activates G protein over β-arrestin signaling while inhibiting NT binding, NT-induced β-arrestin recruitment, and NTSR1 internalization, thereby acting as biased allosteric agonist and negative allosteric modulator. PP-001 also promotes the formation of both homo- and heteromeric multi-receptor complexes. In vivo, PP-001 elicits potent, sustained hypotensive effects, reversible by the NTSR1 antagonist SR48692. Although the precise mechanism of pepducin-receptor interaction remains unclear, we identify a critical N-terminal RKK motif for PP-001’s biological activity. Finally, thermodenaturation assays using purified NTSR1, combined with mutagenesis and molecular docking, provide evidence for the role of the receptor’s H8 domain in direct pepducin interaction. Together, these findings highlight pepducins as versatile modulators of GPCR function and as valuable pharmacological tools for GPCR-targeted drug development.

While transient perioperative side effects of intravenous anesthetics are often tolerated, the persistent postoperative sequelae resulting from drug accumulation pose a critical threat to patient safety. Etomidate, introduced in the 1970s, remains favored for its minimal hemodynamic impact but is severely limited by sustained adrenal suppression, leading to higher mortality and poorer outcomes in critically ill patients. To address these challenges, we reframed our strategy from solely optimizing receptor specificity to enhancing metabolic efficiency, thereby reducing prolonged postoperative exposure and mitigating sustained adverse effects. Using a deep-learning based molecule optimization algorithm, we identified metabolically favorable lead compounds and synthesized 31 novel imidazole-based etomidate derivatives. Among these, ETO-4 emerged as the most promising candidate, retaining potent anesthetic activity while accelerating metabolic clearance and significantly diminishing adrenal suppression. Plasma cortisol assays confirmed the effect of ETO-4 on adrenal function is greatly reduced. These findings underscore a paradigm shift in anesthetic drug design, demonstrating that prioritizing enhanced metabolic profiles can yield safer, more effective agents that improve postoperative outcomes.

Biofilm-mediated resistance in multidrug-resistant (MDR) Pseudomonas aeruginosa infections severely compromise antibiotic efficacy in clinical applications. Antibacterial adjuvants represent a promising strategy to restore antibiotic sensitivity and reduce therapeutic dosages. To identify new antibacterial adjuvants with unique structure and mechanism, we established a generative active learning workflow integrating an in-house compound repository of 725 biofilm inhibitors and a library of potential antibiofilm targets. The most potent compound STY17 was identified with sub-micromolar antibiofilm activity (IC₅₀ = 0.29 ± 0.01 μmol/L). In clinically isolated MDR Pseudomonas aeruginosa, STY17 significantly inhibited biofilm formation, potently synergized with tobramycin and ciprofloxacin, and suppressed the resistance development of these antibiotics. Furthermore, mechanistic studies indicated that STY17 inhibited succinate dehydrogenase to disrupt biofilm formation. In vivo, STY17 significantly enhanced the antibacterial activity of tobramycin and ciprofloxacin in Galleria mellonella and the mouse wound infection model with favorable safety profiles. These findings validated the utility of machine learning to discover novel antibacterial adjuvants, revealing STY17 as a promising candidate for antibacterial adjuvants against MDR Pseudomonas aeruginosa infections.

Enhancing immunity offers a versatile yet effective strategy for treating or preventing diseases. Here, we obtained a new polysaccharide (AFP-80) from Anoectochilus formosanus that unexpectedly stimulated immune systems without causing any detectable adverse effects. AFP-80 was applied to encapsulate Lactobacillus plantarum (LP), a probiotic with immunoregulatory properties, through bridging by Fe³⁺-tannic acid network (Fe-TA) to establish an oral immune enhancer (LP@Fe-TA@AFP-80). Upon oral administration, LP@Fe-TA@AFP-80 colonized intestines with high survival rates, aided by gastrointestinal stress-shielding and adhesive properties of AFP-80 and Fe-TA, respectively, leading to synergistic immuno-enhancing effects through combining AFP-80 and live LP. As a result, in immunocompromised mice, LP@Fe-TA@AFP-80 significantly renovated immune functions, which was deciphered to be closely associated with the rebalance of gut microbiota toward a profile with positively-immunoregulatory bacteria enriched as well as the involvement of peroxisome proliferators-activated receptor signaling pathway activation. Additionally, LP@Fe-TA@AFP-80 effectively treated cancer, e.g., 4T1 breast and MC38 colon tumors, either alone or in combination with immune checkpoint blockade therapy, by remodeling tumor immune microenvironment and gut microbiota. Moreover, LP@Fe-TA@AFP-80 demonstrated excellent biocompatibility, as directly revealed by negligible biotoxicity even after 6 months of consecutive ingestion, highlighting its great potential to be implemented into people’s daily life for disease prevention.

Repolarizing immunosuppressive M2-phenotype tumor-associated macrophages (TAMs) and blocking the CD47/SIRPα axis are promising strategies to enhance cancer immunotherapy. However, non-selective disruption of macrophage phenotypic balance and CD47/SIRPα signaling can lead to immune-related side effects. To address this, we develop a smart biomimetic nanoparticle (PARM) loaded with R848 and manganese ions (Mn²⁺). PARM is coated with an apoptotic vesicle membrane and a pH-sensitive PEG corona, enabling targeted delivery to TAMs in the acidic tumor microenvironment (TME). The PEG corona protects the nanoparticle from uptake during circulation and sheds in the TME, exposing the apoptotic vesicle membrane. This triggers specific recognition and uptake by TAMs via the “eat-me” signal. R848 and Mn²⁺ repolarize TAMs into a pro-inflammatory phenotype, while the activation of cGAS-STING pathway by Mn²⁺ reduces SIRPα expression and enhances TAM phagocytosis. In vivo studies demonstrate that PARM remodels the immunosuppressive TME by repolarizing TAMs and promoting CD8⁺ T cell infiltration. This leads to significant inhibition of tumor growth and metastasis. These findings highlight the multifaceted role of the cGAS-STING pathway in TAM modulation and present a novel strategy for enhancing macrophage-based cancer immunotherapy.

Tailoring tumor-associated macrophages (TAMs) into the tumoricidal phenotype represents a high-profile strategy for tumor immunotherapy. However, the existing TAMs repolarization strategies are restricted by hostile microenvironmental stress and metabolic compensation, leading to limited anti-tumor phenotype sustainability. Herein, a “Spark-Relay” nanoinitiator (SRN) with flexible S/R reactant ratio was meticulously designed for rewiring TAMs as tumoricidal bioreactor and precisely overcome metabolic compensation. Briefly, SRN reshaped TAMs in situ relying on tissue tropism of macrophage membrane, firstly upregulating their reactive oxygen species (ROS) production via burst release of Spark element and initially shifting the TAMs into the anti-tumoral phenotype, subsequently elevating NO production by releasing Relay element, which is converted to NO via reaction with ROS and iNOS, leading to the generation of additional ROS and creating a positive feedback loop, thereby strengthening the metabolic rewiring of TAMs from passively defensive oxidative phosphorylation to positively offensive glycolysis, which significantly enhanced the antitumoral activity of TAMs and shrunk tumor immunosuppressive microenvironment, emerging as 5.5-fold increase of tumor-suppressive/supportive ratio (TSSR) of TAMs, 4.0-fold elevation of CD8⁺ T cell infiltration, contributing to a satisfactory tumoricidal efficacy and providing a cascade amplification mode for TAMs-based cancer therapeutics.

Intestinal barrier disruption, driven by oxidative stress, ferroptosis, immune imbalance, and gut microbiota dysbiosis, plays a crucial role in inflammatory bowel disease (IBD) pathogenesis. Current treatments are often ineffective and cause side effects, emphasizing the need for novel therapies. Here, we have developed an engineered probiotic-derived outer membrane vesicle (OMV), GDO@CM, combining antioxidant gallic acid (GA) and anti-inflammatory H₂S for targeted intestinal barrier repair. Constructed from Escherichia coli Nissle 1917 (EcN)-derived OMVs, GA and diallyl trisulfide (DATS) are incorporated into the hydrophilic inner cavity and lipid bilayer, respectively, while mannose-decorated chitosan (CM) is electrostatically attached to the OMVs surface, enhancing stability and enabling targeted delivery to damaged colonic lesions. GDO@CM efficiently enters activated immune cells and epithelial cells, where GA scavenges reactive oxygen species and inhibits ferroptosis, while H₂S amplifies anti-inflammatory effects. OMVs further synergize with GA and DATS to suppress pathogenic bacteria. These combined actions facilitate effective barrier repair and alleviate IBD symptoms. Single-cell RNA sequencing reveals that GDO@CM reduces inflammation, increases the proportion of reparative M2 macrophages and intestinal stem cells, and promotes epithelial cell proliferation via the APP/CD74 axis. Our findings establish GDO@CM as a promising multi-target therapeutic for IBD, offering a novel strategy for intestinal barrier restoration.

Low-molecular-weight heparin (LMWH) is one of the most clinically used anticoagulants to improve venous circulation. However, LMWH requires repeated injections, leading to poor medication compliance. Herein, we developed a deep eutectic solvents (DESs)@enteric technology, an intestinal absorption promotion platform, for the oral administration of LMWH. DESs with geranic acid (Ge) and choline (Ch) were first synthesized and characterized. Then, the DES-LMWH complex was prepared with a DES₂:₁ (Ge and Ch in a 2:1 molar ratio), demonstrating superior intestinal absorption and high biocompatibility. Finally, DES-LMWH was granulated and loaded into an enteric capsule. In vitro, DESs reversibly opened the tight junctions between intestinal epithelial cells, facilitating the paracellular transport of LMWH. In vivo, the DES-LMWH oral enteric capsule demonstrated an absolute bioavailability of 19.93%, reaching the venous thromboembolism (VTE) prophylaxis threshold and reducing the embolization area by 52.01% in a FeCl₃-induced rat femoral venous thrombosis model. In conclusion, DES@enteric technology effectively enhances the oral absorption of LMWH and holds promising translational potential.

Cardiovascular diseases induced by arterial thrombosis, such as myocardial infarction and ischemic stroke, have gradually emerged as critical threats to human life and health. Sequentially targeted delivery systems are highly desired to be developed to improve the delivery of thrombolytics and anti-inflammatory medications to the site of the thrombus, respectively, to pursue a maximized combinational effect. Herein, we developed a probiotic-platelet hybrid vesicle-targeted drug delivery system (Fer-1@PLevs-C&U) to achieve sequential anti-thrombolytic delivery against thrombus and prevent its recurrence. The hybrid vesicles (PLevs) were prepared by mixing platelet membrane with Lactobacillus plantarum-derived bacterial extracellular vesicles (Levs) and further decorated with DSPE-PEG₂₀₀₀-CREKA, achieving “point-to-point” multi-covalent targeting of thrombus components. After targeted localization to the arterial thrombotic site, Fer-1@PLevs-C&U gradually releases the thrombolytic drug urokinase-type plasminogen activator (UPA) and Ferroptosis inhibitor (Fer-1) to facilitate thrombus dissolution and regulate the inflammatory microenvironment. By enhancing the eNOS expression and reducing the secretion of inflammatory factors TNF-α and IL-6, Fer-1@PLevs-C&U significantly reduced the inflammatory microenvironment and inhibited recurrent thrombus. Therefore, Fer-1@PLevs-C&U constitutes a novel biomimetic drug delivery platform with promising therapeutic potential and practical applicability.