Purpose: While bubble-carrier systems hold great promise for enhancing the oral bioavailability of poorly water-soluble drugs, a deeper understanding of their underlying absorption mechanisms is still required. This study aimed to address this gap by developing a bubble-driven delivery system (BDDS) using the poorly water-soluble andrographolide (AG) as a model drug, and systematically evaluating its solubilization capacity and intestinal permeability. Methods: The BDDS was optimized through the screening of effervescent formulations (citric acid: Na2CO3 = 6:5) and bile salts (sodium deoxycholate, SDC). Its performance was comprehensively assessed via in vitro dissolution tests, an everted intestinal sac assay, Caco-2 cell monolayer experiments, pharmacokinetic studies, and yeast-induced fever rat models. Results: The BDDS achieved 75.0%-88.7% dissolution in gastrointestinal pH media, which was markedly superior to that of commercial dripping pills (DP; 30.5-50.2%). Its apparent permeability coefficient was 7.7-fold and 3.6-fold higher than that of free AG and DP, respectively. Transport studies indicated that SDC enhanced AG permeability primarily via the apical sodium-dependent bile acid transporter (ASBT)-mediated pathway, a mechanism further confirmed by inhibition with linerixibat. The relative oral bioavailability of BDDS versus DP reached 257.1%. BDDS demonstrated enhanced efficacy by significantly inhibiting key pro-inflammatory cytokines and thermoregulatory mediators in yeast-induced fever rats. Discussion: The BDDS effectively overcomes AG's low solubility (by SDC solubilization and CO2 bubbles) and permeability (via ASBT transport), outperforming conventional formulations in terms of preparation simplicity and storage stability. Conclusion: BDDS is a promising strategy to improve oral absorption and therapeutic efficacy of BCS Class IV drugs like AG, with ASBT-mediated transport as a key mechanism.

Klotho is a longevity-associated and tissue-protective protein involved in mineral metabolism, oxidative stress, inflammation, fibrosis, cellular senescence, and neurovascular homeostasis. However, nearly three decades after its discovery, no Klotho-based therapy has been approved, highlighting a translational gap that extends beyond biological validation. This review reframes Klotho translation as a pharmaceutical sciences challenge, focusing on how to convert Klotho into a druggable, manufacturable, deliverable, and clinically controllable therapeutic product. We summarize the isoform-specific properties of membrane-bound α-Klotho, soluble α-Klotho, and β-Klotho that are relevant to product design, and review the current clinical and preclinical landscape dominated by gene-, mRNA-, and antibody-based approaches. We further distinguish confirmed developability barriers, including renal handling and limited systemic persistence, from plausible risks common to macromolecular biologics, such as aggregation, chemical degradation, immunogenicity, and poor tissue penetration. Finally, we evaluate emerging delivery and formulation strategies, including viral and non-viral gene delivery, extracellular vesicles, hydrogels, ultrasound-targeted microbubbles, osmotic pumps, and long-acting protein engineering. An integrated roadmap combining molecular engineering, disease-specific delivery, pharmacokinetic/pharmacodynamic biomarkers, manufacturability assessment, and repeated-dose safety evaluation may help transform Klotho from a promising anti-aging molecule into a clinically viable biologic platform.

ABSTRACT Natural drug‐food homology compounds have vast therapeutic potential, particularly for inflammatory bowel disease (IBD), but their poor water solubility, instability, and limited mucosal availability hinder their clinical translation. As a proof of principle, the natural polyphenol resveratrol (Res) was conjugated to a disulfide‐bridged carboxymethyl chitosan (CMCS) for inclusion in a negatively charged hydrogel (CMTR‐Gel). The CMTR‐Gel was stable, with a zeta potential of approximately −37 mV in the gastrointestinal environment, and significantly inhibited reactive oxygen species production and mitochondrial depolarization in RAW 264.7 cells under lipopolysaccharide‐induced stress. The orally administrated CMTR‐Gel preferentially adhered to inflamed mucosa for 24 h through electrostatic interactions between anionic CMCS and cationic transferrin. Because of its prolonged adhesion and precise Res release at inflamed mucosa, CMTR‐Gel achieved superior therapeutic efficacy compared to the first‐line drug sulfasalazine in colitis mice, and was especially efficacious for mucosal healing through its repair of tight junction integrity. The CMTR‐Gel represents a promising strategy for the application of natural drug‐food homology compounds in the management of IBD.

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by the presence of amyloid-β (Aβ) plaques and neurofibrillary tangles of hyperphosphorylated Tau protein primarily in the hippocampus and cortex, as well as oxidative stress in neuronal tissues. Given the complex pathophysiology of AD, therapeutic strategies that target multiple pathological pathways simultaneously may offer greater efficacy. Here, we designed and synthesized a series of flavonoid–DPCI hybrids that target β-site amyloid precursor protein cleaving enzyme 1 (BACE1), glutaminyl cyclase (QC), and reactive oxygen species (ROS) using a multi-target-directed ligand strategy. These hybrids integrate a flavonoid motif for BACE1 inhibition and ROS scavenging with a 5-methyl-imidazole group for Zn2+ chelation in QC. The compounds exhibited potent activity against all three targets; the representative lead compound 30 showed balanced inhibitory potency with IC₅₀ values of 8.76 μM against BACE1 and 2.25 μM against QC, along with favorable antioxidant capacity. Structure–activity relationship and molecular docking analyses confirmed synergistic interactions between the compounds and their targets. These findings suggest that flavonoid–DPCI hybrids are promising multifunctional inhibitors offering a novel treatment strategy for AD.

Pancreatic ductal adenocarcinoma (PDAC) is difficult to treat due to a dense stromal barrier and an immunosuppressive tumor microenvironment (TME), driven in part by overexpression of isoQC and DPP4. IsoQC promotes CD47-SIRPα-mediated immune evasion, while DPP4 contributes to stromal remodeling and limits T-cell infiltration. Targeting either factor alone is insufficient. Here, we developed a dual-targeting nanoplatform, iRGD@4, composed of iRGD-functionalized polymeric micelles loaded with a DPP4-responsive prodrug of an isoQC inhibitor. The system enables tumor-specific activation, as elevated DPP4 in PDAC cleaves the prodrug to release the active inhibitor locally. Nanoparticles showed uniform size (158.2 nm), good stability (−29.5 mV), and high drug loading (10.6%). In vitro, the prodrug was efficiently activated by DPP4, releasing an inhibitor with IC50 of 0.68 μM. iRGD@4 enhanced cellular uptake and achieved Golgi localization. In vivo, it showed ∼10-fold higher tumor accumulation than free drug and significantly inhibited tumor growth. Mechanistically, it reduced pE-CD47 expression by 62.3%, increased macrophage infiltration, and reshaped cytokine profiles. The system exhibited good biocompatibility with minimal toxicity. Overall, iRGD@4 effectively overcomes stromal and immune barriers in PDAC through enzyme-activated, tumor-targeted dual modulation.

This study uses Scientometric analysis to review Sigesbeckiae Herba (SH) research worldwide from 1980 to 2024. Data-driven analysis identifies patterns, partnerships, and possibilities. Due to its wide profile and traditional medical uses, SH is gaining attention, according to this research. China has made the most important contributions to SH research, followed by the United stated of America and Korea. Countries and organizations have not collaborated on SH research. We found “anti-inflammation,” “rheumatic arthritis,” and “NF-κβ” using keyword co-occurrence analysis. Citation analysis showed SH research's most prominent institutions, journals, and authors, offering a complete snapshot of organization and diffusion. The data shows that this subject has received substantial academic interest despite a reduction in publications since 2019. These findings highlight the need for cooperation, multidisciplinary methods, and attention to underrepresented disciplines for academics, funding organizations, and policymakers. SH has been used to cure inflammation. Sigesbeckia orientalis L., S. pubescens Makino, and S. glabrescens Makino are the primary botanical sources. This research extensively evaluates SH literature, focusing on its botanical properties, bioactive components, pharmacological effects, and probable toxicity. More than three hundred phytochemicals, including flavonoids, diterpenoids, sesquiterpenoids, oxylipins, and lignoids, were identified in the extract, many of which are known to exert diverse biological effects. The extracts of SH are used in traditional Chinese medicine (TCM) due to its immunomodulatory, anti-inflammatory, and anti-cancer activities and are being considered for cosmetics and skin issues. Pharmacological benefits of diterpenoids, sesquiterpenoids, and flavonoids are found. The study recommends further research and cooperation to enhance TCM and herbal research by comparing the chemical and pharmacological properties of these three SH species.

Upregulated glutaminyl cyclase isoenzyme (isoQC) contributes to cancer development by catalyzing pE-CD47 generation and thus enhancing CD47-SIRPα binding and subsequent "don't eat me" signals. We thus consider that isoQC could represent a novel target for cancer therapy. We previously prepared a series of diphenyl conjugated imidazole derivatives (DPCIs) and evaluated their use as glutaminyl cyclase (QC) inhibitors. Here, a new series of DPCIs was rationally designed and synthesized. As anticipated, the analogues exhibited considerably improved inhibitory potency against both QC and isoQC. Crucially, these chemicals exhibited marked selectivity toward isoQC. Further assessments established that one selected compound (27) did not affect the viability of A549, H1299, PC9, or HEK293T cells or the body weight of mice. This compound did, however, reduce pE-CD47 levels in infected A549 cells (isoQC_OE and isoQC_KD) and exhibited apparent anti-cancer effects in vivo by downregulating the level of pE-CD47 via the inhibition of isoQC activity. Taken together, these findings indicated that the compounds synthesized in this study could represent potential QC/isoQC inhibitors for the treatment of cancers.

Efficient cytosolic delivery of small interfering RNA (siRNA) remains a challenge in RNA therapeutics, particularly for oral administration. Identifying new cytosolic stimulus-responsive targets is thus crucial for optimizing siRNA delivery. Given the elevated intracellular arginase-1 (Arg1) levels in inflamed colonic mucosa, we developed lipid nanoparticles (siRNA-LANPs) derived from L-arginine-modified chitosan (ACS) for the cytosolic release of TNF-alpha-siRNA to treat ulcerative colitis (UC). The siRNA-LANPs exhibited superior Arg1-binding affinity compared to its substrate, L-arginine, enabling rapid Arg1-responsive release. Moreover, we found that siRNA-LANPs maintained nanoparticle stability, and 84.5 % of the loaded siRNA remained intact in simulated digestive fluids over 12 h. Following oral administration, siRNA-LANPs efficiently penetrated the mucus layer and preferentially accumulated in inflamed colonic tissue compared to healthy colon tissue. In DSS-induced colitis mice, siRNA-LANPs significantly reduced disease severity through TNF-alpha silencing, achieving greater therapeutic efficacy than sulfasalazine (a first-line UC treatment). The siRNA-LANPs exhibited superior TNF-alpha silencing compared to transfection reagent Lipo3000 and TNF-alpha biologic agent infliximab. The enhanced performance was attributed to multiple endocytic pathways, improved lysosomal escape capability (29.7 %), and Arg1-mediated cytoplasmic siRNA release. In summary, Arg1 seems to be a promising intracellular target for stimulus-responsive, oral siRNA delivery in UC treatment.

Dipyridamole (DIP) has shown promising effectiveness in treating Inflammatory Bowel Disease (IBD), especially in pediatric populations. However, existing formulations are unsuitable for children due to poor solubility and swallowing difficulties. This research aims to develop a novel DIP formulation with improved dissolution rate and bioavailability, tailored for children, using computational and experimental methods. Molecular dynamics modeling was employed to identify the optimal crystal inhibitor for DIP, with Soluplus emerging as the best candidate. This result was further validated through dissolution tests. A dry nanosuspension was then prepared using a wet milling approach followed by freeze-drying. The optimal formulation was evaluated in various dissolution media, resulting in significant improvements. Characterization techniques such as Dynamic Light Scattering (DLS), Powder X-ray Diffraction (PXRD), and Field Emission Scanning Electron Microscopy (FESEM) confirmed the crystalline state and particle size of DIP in the optimal formulation. Subsequent cell and animal studies demonstrated that the optimal formulation outperformed both the commercial product and pure DIP. The absolute bioavailability of pure DIP, commercial tablets, and optimal formulation was 9.51%, 18.17%, and 42.15%, respectively. Physiologically based pharmacokinetic (PBPK) modeling was then utilized to predict and evaluate the in vivo behavior for both adult and pediatric populations, showing good performance in both groups. In conclusion, our study successfully developed a DIP dry nanosuspension with improved dissolution and bioavailability, specifically tailored for pediatric use. The combination of computational and experimental methods offers a strong foundation for future formulation development, significantly reducing both time and costs.

BackgroundUlcerative colitis (UC) is a prevalent immune-mediated inflammatory bowel disease characterized by mucus secretion, hematochezia, and diarrhea. This study compared the therapeutic effects of three Siegesbeckiae Herba (SH) species used in traditional Chinese medicine—Sigesbeckia orientalis L (SO), Sigesbeckia pubescens Makino (SP), and Sigesbeckia glabrescens Makino (SG) — in dextran sulfate sodium (DSS)-induced UC mice.MethodsUC was induced in C57BL/6 mice with 3% DSS for 7 days. Cytokine levels in serum and colon tissues were measured by enzyme-linked immunosorbent assay. Protein and gene expression were analyzed using Western blotting and PCR. Histopathological changes were assessed via hematoxylin-eosin staining, immunohistochemistry, and immunofluorescence. Fecal specimens were collected for gut microbiota analysis. An in vitro UC model was also established in NCM460 cells using lipopolysaccharide (LPS), and Caco-2 cells were used to examine intestinal mucosal integrity.ResultsSP substantially decreased the disease activity index, enhanced colon shortening, and mitigated histological damage in comparison to the model group. Mechanistic investigations demonstrated that SP functioned via the activation of the Nrf2/Keap1 pathway, markedly increased the activity of the antioxidant enzyme glutathione in colon tissues, decreased the concentration of the oxidative marker malondialdehyde, and upregulated the expression of the downstream genes H O -1 and NQO1.ConclusionThe study reveals for the first time the differences in efficacy of different species of SH and its molecular mechanism, demonstrating that SP increases oxidative defense via the activation of the Nrf2/Keap1 pathway, therefore mitigating colitis and oxidative damage in UC mice. This discovery not only establishes a scientific foundation for the selective preference of SH species but also offers a novel technique for the creation of natural pharmaceuticals aimed at the Nrf2 pathway for the treatment of UC.

Klotho (KLO) is an anti-fibrotic protein expressed in the kidneys and has been decreasing in the development of renal fibrosis (RF). However, restoring the decline in KLO levels remains a great challenge during RF treatment. Herein, an injectable KLO-loaded chitosan (CS) hydrogel (KLO-Gel) is designed to achieve localized and prolonged release of KLO in the RF treatment. KLO-Gel was prepared by cross-linking CS with β-glycerophosphate (β-GP), followed by rapid (within 3 min) thermosensitive gelation at 37 °C. Furthermore, KLO-Gel exhibited a slow and sustained release (over 14 d) of KLO both in PBS and in the kidneys of mice with unilateral ureter obstruction (UUO). A single local injection of KLO-Gel into the renal capsule of UUO mice was more effective at reducing RF (i.e., maintaining renal function and tissue structure, alleviating extracellular matrix accumulation, and inhibiting the TGF-β1/Smad2/3 signaling pathway) over a 14-d period than daily intraperitoneal injections of free KLO or captopril. Crucially, CS was found to induce endogenous KLO secretion, highlighting the added value of using CS in RF treatment. Overall, this study demonstrated that KLO-Gel enhanced the anti-fibrotic efficacy of KLO while minimizing its off-target toxicity, and its clinical potential awaits further validation.

Intestinal mucosal barrier loss is responsible for the chronic and recurrent ulcerative colitis. Myosin light chain kinase (MLCK) is a potential therapeutic target of the intestinal mucosal barrier dysfunction. Here, we developed a reactive oxygen species (ROS)-sensitive hydrogel (ATG-CS-Gel) derived from a diselenide-bridged arctigenin (ATG) and chitosan (CS) conjugate, with the aims of targeting to inflamed mucosa and modulating MLCK. Our results demonstrated that ATG-CS-Gel achieved ROS-responsive release and significantly inhibited ROS production and mitochondrial depolarization in the Caco-2 and HT-29/MTX-E12 cells under H2O2-induced stress conditions. Compared with normal tissues, orally-administrated ATG-CS-Gel preferentially adhered to the inflamed mucosa for 24 h, which was attributed to the adhesion between CS and mucin. Therapeutically, ATG-CS-Gel reduced inflammatory symptoms, accelerated intestinal mucosal healing, scavenged excessive ROS, reshaped intestinal flora, and eventually achieved much better therapeutic efficacy in DSS-induced colitis mice when compared to 5-aminosalicylic acid. Moreover, ATG-CS-Gel was demonstrated to reverse intestinal mucosal barrier loss by blocking MLCK activation and maintaining tight junction expression. In summary, this study highlights the potential of MLCK modulation in the restoration of intestinal mucosal barrier using ATG-CS-Gel. The development of ATG-CS-Gel represents a novel and promising strategy for the treatment of ulcerative colitis.

Alzheimer's disease (AD) is a major cause of dementia and one of the most common chronic diseases affecting the aging population. Because AD is considered a public health priority, there is a critical need to discover novel and effective agents for the treatment of this condition. In view of the known contribution of up-regulated glutaminyl cyclase (QC) and glycogen synthase kinase-3 beta (GSK-3 beta) to the initiation of AD, we previously evaluated a series of dual inhibitors containing maleimide and imidazole motifs as potential anti-AD agents. Here, we assessed another series of hybrids containing maleimide and imidazole motifs to gain an in-depth understanding of the structure-activity relationship (SAR). Based on the primary screening, the introduction of 5-methyl imidazole at one side of the molecule did not enhance the QC-specific inhibitory activity of these hybrids (2, IC50 = 1.22 mu M), although the potency was increased by 2 ' substitution on the maleimide motif at the other side of the molecule. Interestingly, compounds containing 5-methyl imidazole exhibited stronger GSK-3 beta-specific inhibitory activity (2, IC50 = 0.0021 mu M), and the electron-withdrawing group and 2 ' and 3 ' substitution were favorable. Further investigation of substitutions on the maleimide motif in compounds 14-35 revealed that QC-specific inhibition in the presence of piperidine was improved by introduction of a methoxy group (R2). Increasing the linker length and introduction of a methoxy group (R2) also increased the GSK-3 beta-specific inhibitory potency. These findings were further confirmed by molecular docking analysis of 33 and 24 with QC and GSK-3 beta. Overall, these hybrids exhibited enhanced inhibitory potency against both QC and GSK-3 beta, highlighting an important strategy for improving the potency of hybrids as dual-targeting anti-AD agents.

There is increasingly keen interest in developing orally delivered targeted drugs, especially for diseases that require long-term medication. Hence, we manufactured nanoparticles derived from methoxypolyethylene glycol-chitosan (PCS) to enhance the oral delivery and kidney-targeted distribution of salvianolic acid B (SalB), a naturally occurring renoprotective and anti-fibrotic compound, as a model drug for the treatment of renal fibrosis. Orally administered SalB-loaded PCS nanoparticles (SalB-PCS-NPs) maintained good stability in the gastrointestinal environment, improved mucus-penetrating capacity, and enhanced transmembrane transport through a Caco-2 cell monolayer. The relative oral bioavailability of SalB-PCS-NPs to free SalB and SalB-loaded chitosan nanoparticles (SalB-CS-NPs) was 367.0 % and 206.2 %, respectively. The structural integrity of SalB-PCS-NPs after crossing the intestinal barrier was also validated by Förster resonance energy transfer (FRET) in vitro and in vivo. Fluorescein isothiocyanate (FITC)-labeled SalB-PCS-NPs showed higher kidney accumulation than free FITC and FITC-labeled SalB-CS-NPs (4.6-fold and 2.1-fold, respectively). Significant improvements in kidney function, extracellular matrix accumulation, and pathological changes were observed in a unilateral ureter obstruction mouse model of renal fibrosis after once daily oral treatment with SalB-PCS-NPs for 14 days. Thus, oral administration of SalB-PCS-NPs represents a promising new strategy for kidney-targeted drug delivery.

Drug nanocrystal engineering is an attractive pharmaceutical approach to enhancing the oral bioavailability of poorly soluble drugs. The mechanism of drug nanocrystal stabilization, however, is unclear. Here we developed andrographolide nanocrystals (AG-NCs) with various nonionic surfactants (Pluronic-F127, TPGS, or Brij-S20). We detected AG micelles (AG-MCs) at an andrographolide to nonionic surfactant ratio of 10:10 (w/w) and poor AG-NC size stability. We thus quantified the unbound Pluronic-F127 in AG-NCs and found that the proposed instantaneous binding rate sharply declined with increasing Pluronic-F127 input. We determined that the saturation dose of TPGS on AG-NCs was approximately 10:10 (w/w) and recommend it as a key criterion for nanocrystal formulation. Although AG-NCs exhibited a marginally faster dissolution rate, they possessed better mucus-penetrating and transmembrane transport capacities and significantly enhanced oral absorption compared to AG-MCs. These findings give insights into the impact of a stabilizer during the preparation process and the oral absorption of drug nanocrystals.

To understand how upregulated isoglutaminyl cyclase (isoQC) is involved in the initiation of diseases such as cancer, we developed a human KYSE30 carcinoma cell model in which isoQC was stably overexpressed. GO and KEGG analysis of the DEGs (228) and DEPs (254) respectively implicated isoQC on the proliferation invasion and metastasis of cells and suggested that isoQC might participate in the regulation of MAPK, RAS, circadian rhythm, and related pathways. At the functional level, isoQC-overexpressing KYSE30 cells showed enhanced proliferation, migration, and invasion capacity. Next, we decided to study the precise effect of isoQC overexpression on JNK, p-JNK, AKT, p-AKT, ERK, p-ERK, and PER2, as RNA levels of these proteins are significantly correlated with signal levels indicated in RNA-Seq analysis, and these candidates are the top correlated DEPs enriched in RT-qPCR analysis. We saw that only p-ERK expression was inhibited, while PER2 was increased. These phenotypes were inhibited upon exposure to PER2 inhibitor KL044, which allowed for the restoration of p-ERK levels. These data support upregulated isoQC being able to promote cancer cell proliferation and migration in vitro, likely by helping to regulate the MAPK and RAS signaling pathways, and the circadian protein PER2 might be a potential mediator.

Glutaminyl cyclase (QC) plays a crucial role in the early stages of Alzheimer's disease (AD), thus inhibition of QC may be a promising strategy for the treatment of early AD. Therefore, QC inhibitors with novel chemical scaffolds may contribute to the development of additional anti-AD agents. We conducted a virtual screening of 3 million compounds from the Chemdiv and Enamine databases, to discover potential scaffolds for QC inhibitors. Three scaffolds, 120974, 147706, and 141449, were selected from this structure-based virtual screening through a combination of pharmacophore modeling, a receptor-ligand pharmacophore model, and the GALAHAD model, and furtherly filtered by chelation with zinc ion and docking properties. Consequently, three compounds, 1, 2, and 3, were designed and synthesized based on these three scaffolds, respectively. The IC50 of compounds 1 and 3 against QC were 14.19 +/- 4.21 and 4.34 +/- 0.35 mu M, respectively. Our results indicate that the new scaffolds selected using a virtual screening process exhibit potential as novel QC inhibitors.

The blood-brain barrier (BBB) prevents pathogens and toxins in the bloodstream from reaching the brain, but also inhibits the delivery of agents intended to treat central nervous system disorders, such as Alzheimer's disease (AD). In this study, we prepared and evaluated a novel nano-delivery vehicle system composed of lactoferrin-conjugated (Lf-PIC@Se) micelles. We used a COOH-PEG-PAsp-PV@Se synthesis-based method to prepare the micelles, which involved self-assembly followed by EDC-NHS coupling. Using glutaminyl cyclase inhibitor 8 as a model encapsulated chemical, Lf-PIC@Se micelles achieved a good loading capacity. In vitro analysis demonstrated that Lf-PIC@Se/8 micelles were stable in both neutral and acidic pH solutions in the presence or absence of H2O2, and confirmed their biosafety and compatibility in PC12 and bEND.3 cells. Notably, the cell uptake of Lf-PIC@Se/C6 micelles was much higher than that of PIC@Se micelles, and occurred through LfR-mediated endocytosis. The presence of Se meant that Lf-PIC@Se micelles acted as ROS scavengers in PC12 cells under H2O2-induced oxidative stress, which inhibited oxidative damage and increased mitochondrial membrane potential. Hemolysis assays further demonstrated that Lf-PIC@Se represent a biocompatible carrier. Finally, in vivo experiments in mice suggested that Lf-PIC@Se micelles successfully crossed the BBB, confirming their potential as vehicles for drug delivery when treating AD and other central nervous system disorders.

Alzheimer's disease (AD), multifactorial disease, is recognized as one of the most common forms of dementia, and the efficacy of anti-AD drugs is limited clinically. Up-regulated glutaminyl cyclase (QC) and glycogen synthase kinase-3β (GSK-3β) have been identified as two critical elements involved in AD recently. Here, a series of novel chemicals containing maleimide and imidazole motif were designed and synthesized as dual inhibitors targeting QC and GSK-3β. Based on primary screening, compound 2 (2.26 μM), 5 (2.37 μM), 8 (1.34 μM), 21 (2.44 μM), 25 (0.36 μM), 27 (1.76 μM), 28 (1.04 μM), 33 (2.08 μM) and 34 (2.33 μM) exhibited notable human QC (hQC) inhibitory potency, while compound 1 (0.014 μM), 7 (0.04 μM), 8 (0.057 μM), 19 (0.034 μM), 24 (0.014 μM), 32 (0.032 μM), 38 (0.051 μM), 39 (0.044 μM), 44 (0.048 μM), 47 (0.011 μM), 49 (0.021 μM) and so on showed remarkable GSK-3β inhibitory activities. And as expected, these chemicals possessed significant inhibitory potency on both hQC and GSK-3β, such as compound 1 (2.80 and 0.014 μM), 8 (1.34 and 0.057 μM), 25 (0.36 and 0.15 μM), 27 (1.76 and 0.069 μM), 28 (1.04 and 0.090 μM), 33 (2.08 and 0.19 μM), 34 (2.33 and 0.11 μM), 35 (2.55 and 0.14 μM), 36 (2.34 and 0.11 μM), etc. Subsequent in vivo studies demonstrated that compound 8 attenuated cognitive deficits and decreased the anxiety-like behavior in 3 × Tg-AD mice. The treatment decreased both pE-Aβ and Aβ accumulation by inhibiting the activity of QC, and decreased the hyperphosphorylation of Tau by reducing the levels of GSK-3β in the brains of AD mice. Results obtained in this research suggested that these novel compounds could be supposed as potential anti-AD agents targeting QC and GSK-3β.