DP-128 is a multitarget benzonaphthyridine-6-chlorotacrine hybrid molecule with potent in vitro anticholinesterase and Aβ42 and tau anti-aggregating activity. While often used as a reference protein aggregation inhibitor, its further development as an anti-Alzheimer agent is limited by significant cytotoxicity, suboptimal aqueous solubility and microsomal stability. Since these drawbacks might arise from its rather high lipophilicity, in this work we have developed a series of more polar analogues, designed by structural modifications at the benzonaphthyridine or 6-chlorotacrine moieties or within the eight-atom linker. Half of the new analogues are indeed slightly more soluble and clearly less cytotoxic than DP-128, display single-digit acetylcholinesterase inhibitory activity, and retain the Aβ42 and tau anti-aggregating potency of the lead, as well as favourable brain permeation and high plasma stability. While further optimization of microsomal stability is necessary for a potential therapeutic use of this class of compounds, hybrids 16 and 17, with similar or even higher Aβ42 and tau anti-aggregating activity and lower cytotoxicity than DP-128, might represent novel pharmacological tools for protein aggregation studies.

The circadian clock mechanism generates 24-h rhythms crucial for regulating various physiological processes, and its dysregulation has been implicated in numerous diseases. In cells, the circadian clock operates through a transcriptional-translational feedback loop, where phosphorylation plays a pivotal role in maintaining accurate circadian rhythms. Consequently, kinase inhibitors have emerged as promising targets for modulating the circadian clock and potentially treating circadian-related diseases. This review aims to provide an overview of the current state-of-the-art of kinase inhibitors with effects on the mammalian circadian clock. By highlighting promising targets and addressing the limitations of existing inhibitors, this review intends to provide a guide for future research efforts towards the development of novel compounds for the treatment of circadian-related disorders. Furthermore, it highlights the critical discrepancy between in vitro activity and in vivo effectiveness, emphasizing the critical need for rigorous in vivo validation to translate the therapeutic potential of kinase inhibitors into effective treatments for these disorders.

In this study, a series of tacrine-celecoxib hybrid compounds was designed and synthesized to modulate key molecular targets implicated in metabolic dysfunction-associated mild cognitive impairment, including cyclooxygenase-2 (COX-2), carbonic anhydrases, and cholinesterases. Among the investigated derivatives, compound 10b was identified as the most balanced multitarget candidate, exhibiting selective inhibition of human butyrylcholinesterase (BuChE, IC50: 470 nM), potent COX-2 inhibitory activity (IC50: 50 nM), and low nanomolar Ki towards human carbonic anhydrase IX (hCA IX). Importantly, 10b showed lower neurotoxicity in neuronal cell models compared with tacrine and staurosporine. In a high-fat diet (HFD)-induced rat model of metabolic dysfunction and cognitive impairment, administration of 10b either orally or via intranasal nanoformulation significantly improved hippocampus-dependent cognitive performance as assessed by a composite behavioral z-score. Administration of 10b was also associated with improvements in glucose homeostasis, insulin sensitivity, lipid profile, together with a reduction of HFD-induced hepatological stress. Consistent with its multitarget profile, biochemical analyses showed significant attenuation of oxidative stress and neuroinflammatory markers in the hippocampus. Finally, docking and molecular dynamics simulation studies provided a structural rationale for the observed in vitro activities of 10b, highlighting favorable binding modes within the active sites of COX-2, BuChE, and hCA IX.

A cost-effective surface plasmon resonance (SPR)-based sensing platform was developed to evaluate alterations in albumin binding capacity under clinically relevant conditions. This ex vivo approach enables real-time assessment of albumin-ligand interactions using albumin directly isolated from plasma, thus overcoming key limitations of conventional in vitro approaches. The sensing surface was prepared by covalently immobilizing a polyclonal anti-albumin antibody onto a CM5 chip, followed by a single-step immunocapture of albumin from patient plasma samples. Mass spectrometry confirmed the selective retrieval of both native and structurally modified albumin forms, preserving their relative abundance and disease-associated microheterogeneity. The sensing surface demonstrated high reusability and analytical reproducibility over ~ 500 capture-release cycles, significantly lowering per-sample costs. Functional validation was performed using ligands targeting the three main albumin binding sites. As proof of application, the system was used to investigate albumin binding properties in plasma from (i) type 2 diabetic patients with (n = 10) and without (n = 10) moderate kidney impairment, and (ii) patients with cirrhosis and acute-on-chronic liver failure (n = 6), a condition associated with extensive albumin damage. The proposed approach provides a robust analytical framework for the functional characterization of circulating albumin in healthy and diseased conditions.

BACKGROUND AND PURPOSE:The rising prevalence of type 2 diabetes mellitus (T2D) has led to an increase in complications, including mild cognitive impairment. Accordingly, there is a growing interest in the potential neuroprotective benefits of newer anti-diabetic drugs, such as dipeptidyl peptidase 4 inhibitors (DPP-4is) and sodium-glucose cotransporter-2 inhibitors (SGLT2is). Although clinical studies suggest that these drugs mitigate cognitive decline, the underlying mechanisms remain unclear. This study aimed to elucidate the potential mechanisms through which DPP-4is or SGLT2is, in combination with metformin, confer neuroprotection. EXPERIMENTAL APPROACH:We conducted a cross-sectional study involving T2D patients on either metformin alone or in combination with a DPP-4i or SGLT2i, alongside healthy controls. Cognitive and metabolic phenotypes were assessed, followed by serum proteomic profiling, computational drug target prediction, network analyses and molecular docking to identify signalling pathways linked to cognitive impairment. KEY RESULTS:T2D patients on combination therapy demonstrated better cognitive function, independent of other phenotypic, metabolic and biochemical factors. Proteomic profiling revealed 36 differentially expressed proteins that were preserved or restored to control levels in the combination therapy group. Gene set enrichment analysis highlighted the modulation of the complement pathway, particularly the involvement of ficolin-3 (FCN3). Molecular docking suggested that the sugar-like or glycyl moieties in anti-diabetic drug molecules interacted with FCN3, potentially inhibiting complement system activation. Such interaction was confirmed by binding studies using surface plasmon resonance. CONCLUSION AND IMPLICATIONS:These findings are significant in tailoring T2D treatment to reduce cognitive complications and exploring drug design to target neuroinflammatory disorders.

Immobilized enzyme reactors (IMERs) are emerging as important components of advanced analytical workflows, in which selective enzymatic transformations are directly combined with continuous-flow operation and high-resolution separation techniques. By converting traditionally offline biochemical reactions into automated, residence-time-controlled modules, IMERs offer major advantages in speed, reproducibility, and compatibility with multidimensional chromatographic and mass spectrometric platforms.In protein analysis, selected IMER-based technologies have reached substantial technical maturity and are increasingly integrated into liquid-chromatography mass spectrometry workflows for peptide mapping, structural characterization, and glycoanalysis, while their routine implementation in regulated quality control remains more limited. In contrast, applications to nucleic acid therapeutics, including oligonucleotides, DNA- and messenger RNA (mRNA)-based modalities, are still at an earlier stage of development but are progressing rapidly. Recent studies highlight growing potential in controlled nuclease processing, online mapping workflows, reusable in vitro transcription platforms, and impurity-oriented analytical strategies. This emerging area is particularly timely because the rapid expansion of nucleic acid medicines is creating analytical demands that align closely with the modular strengths of IMER systems.By examining mature protein applications alongside emerging nucleic acid workflows, this review provides a comparative perspective on how IMER technology is evolving across different classes of macromolecular therapeutics. Current limitations, translational barriers, and future opportunities toward standardized, automated, and data-rich analytical workflows are critically discussed.

In light of the significant correlation between inflammatory alterations and metabolic dysfunction throughout different stages of metabolic disease progression, we focused on utilizing our previously characterized glitazone-derived anti-inflammatory 1,2,3-triazoles as lead compounds to create new multitarget directed ligands that interact with COX-2, peroxisome proliferator-activated receptor γ (PPARγ), and CA within the framework of metabolic disorders. Notably, seven compounds exhibited equivalent or similar COX-2 inhibitory effects to celecoxib. Four compounds, namely, 3b, 3e, 5e, and 5h, exhibited substantial nanomolar inhibitory effects against hCA I, II, IV, and IX isoforms (K i 8.5-833, 0.37-24.6, 44.2-777, and 27.3-32.1 nM, respectively). Furthermore, compounds 5e and 5h demonstrated a significant increase in glucose uptake in the rat hemidiaphragm experiment, outperforming pioglitazone. A robust PPARγ agonism in luciferase assay, full-length human PPARγ transactivation without artificially increasing its expression, and isothermal titration calorimetry for K d determination were used to substantiate their PPARγ-dependent insulin-sensitizing activity. In vivo pharmacokinetic and tissue distribution experiments were carried out, revealing favorable properties. The in vitro activities were reflected into effective in vivo anti-inflammatory potential in the formalin-induced rat paw edema assay, and they also exhibited a favorable ulcerogenic profile. Furthermore, computational target prediction and network pharmacology analysis for the two most active molecules, 5e and 5h, identified important biological pathways associated with the intended outcomes. In this regard, 5e and 5h not only mitigated hyperglycemia and insulin resistance in an in vivo rat model of type 2 diabetes but also protected against renal and lipemic damage caused by metabolic dysfunction. Finally, docking simulations indicated potential binding interactions with the intended biological targets.

Given the urgent need for developing new therapeutic strategies against chronic diseases such as neurodegeneration and cancer, and the evidence of shared biochemical pathways linking Alzheimer's disease to certain types of cancer, we have designed a novel family of multifaceted compounds to target both diseases. For that purpose, isofagomine (a relevant azasugar) - coumarin hybrids were prepared and tested in vitro as potential dual-action molecules. Key structural variations, including different hydroxyl group substitutions and changes in the length of the hydrocarbon linker, had minimal impact on cholinesterase inhibition. All compounds exhibited strong inhibition of butyrylcholinesterase (BuChE), representing the predominant cholinesterase in moderate-to-advanced stages of Alzheimer's disease, with IC50 values in the single-digit micromolar concentration range. Additionally, the isofagomine-coumarin hybrids displayed remarkable selectivity, up to 177-fold, for human BuChE over human acetylcholinesterase (AChE). Docking simulations predicted derivatives to be accommodated within the BuChE binding region. Additionally, the compounds showed reduced neurotoxicity and moderate neuroprotection. Furthermore, a direct correlation was observed between tether length and antiproliferative activity, with the lead compound exhibiting potent effects in the low-micromolar range. 3D Holotomographic microscopy, through continuous live-cell imaging, proved mitotic arrest followed by apoptotic events to be involved in their mode of action. Azasugar-coumarin hybrids constitute promising multifaceted molecules in terms of therapeutics or prevention of neurodegeneration and cancer.

Neurodegenerative diseases currently represent one of the most serious health pitfalls for the world population. Considering their multifactorial nature, research has focused on the study of small molecules able to simultaneously tackle different targets involved in their onset and progression. In this paper, two sets of acylaminopyrazole-based compounds were designed to exploit the aminopyrazole core as a privileged structure properly decorated with an acyl moiety and a further amide function, connected with a proper spacer. Indeed, acylated aminopyrazoles could be able to establish the appropriate hydrogen bond pattern to both bind GSK-3β, responsible for tau hyperphosphorylation, prevent the formation of insoluble Aβ-protein aggregates and have the structural features to show chelating properties towards metals involved in neuroinflammation. The collection of compounds was tested in vitro for GSK-3β inhibition activity, antiaggregating and chelating properties. Selected compounds were able to inhibit GSK-3β in the low micromolar range with a reversible and competitive mechanism of action, as established by Microfluidic Mobility Shift Assay (MMSA) and showed metal chelating ability. Preliminary Structure Activity Relationships (SARs) to hit these distinct and interconnected targets for neuromodulation were established. Finally, selected compounds showed good apparent permeability values in parallel artificial membrane permeability assay (PAMPA) together with good cellular safety profile. The collected results validated acylaminopyrazole as promising scaffold for the development of multitarget-directed ligands. Compounds 1c and 4c emerged as promising prototypes, and deserve further optimization in the search for drug candidates for polypharmacological approach in neurodegenerative disease.

A series of tacrine-donepezil hybrids were synthesized as potential multifunctional anti-Alzheimer's disease (AD) compounds. For this purpose, tacrine and the benzylpiperidine moiety of donepezil were fused with a hydrazone group to achieve a small library of tacrine-donepezil hybrids. In agreement with the design, all compounds showed inhibitory activity toward both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) with IC50 values in the low micromolar range. Kinetic studies on the most potent cholinesterase (ChE) inhibitors within the series showed a mixed-type inhibition mechanism on both enzymes. Also, the docking studies indicated that the compounds inhibit ChEs by dual binding site (DBS) interactions. Notably, tacrine-donepezil hybrids also exhibited significant neuroprotection against H2O2-induced cell death in a differentiated human neuroblastoma (SH-SY5Y) cell line at concentrations close to their IC50 values on ChEs and showed high to medium blood-brain barrier (BBB) permeability on human cerebral microvascular endothelial cells (HBEC-5i). Besides, the compounds do not cause remarkable toxicity in a human hepatocellular carcinoma cell line (HepG2) and SH-SY5Y cells. Additionally, the compounds were predicted to also have good bioavailability. Among the tested compounds, H4, H16, H17, and H24 stand out with their biological profile. Taken together, the proposed novel tacrine-donepezil scaffold represents a promising starting point for the development of novel anti-ChE multifunctional agents against AD.

BACKGROUND:SMYD3 has been found implicated in cancer progression. Its overexpression correlates with cancer growth and invasion, especially in gastrointestinal tumors. SMYD3 transactivates multiple oncogenic mechanisms, favoring cancer development. Moreover, it was recently shown that SMYD3 is required for DNA restoration by promoting homologous recombination (HR) repair.METHODS:In cellulo and in vivo models were employed to investigate the role of SMYD3 in cancer chemoresistance. Analyses of SMYD3-KO cells, drug-resistant cancer cell lines, patients' residual gastric or rectal tumors that were resected after neoadjuvant therapy and mice models were performed. In addition, the novel SMYD3 covalent inhibitor EM127 was used to evaluate the impact of manipulating SMYD3 activity on the sensitization of cancer cell lines, tumorspheres and cancer murine models to chemotherapeutics (CHTs).RESULTS:Here we report that SMYD3 mediates cancer cell sensitivity to CHTs. Indeed, cancer cells lacking SMYD3 functions showed increased responsiveness to CHTs, while restoring its expression promoted chemoresistance. Specifically, SMYD3 is essential for the repair of CHT-induced double-strand breaks as it methylates the upstream sensor ATM and allows HR cascade propagation through CHK2 and p53 phosphorylation, thereby promoting cancer cell survival. SMYD3 inhibition with the novel compound EM127 showed a synergistic effect with CHTs in colorectal, gastric, and breast cancer cells, tumorspheres, and preclinical colorectal cancer models.CONCLUSIONS:Overall, our results show that targeting SMYD3 may be an effective therapeutic strategy to overcome chemoresistance.

Introduction: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer still lacking effective treatment options. Chemotherapy in combination with immunotherapy can restrict tumor progression and repolarize the tumor microenvironment towards an anti-tumor milieu, improving clinical outcome in TNBC patients. The chemotherapeutic drug paclitaxel has been shown to induce immunogenic cell death (ICD), whereas inhibitors of the indoleamine 2,3- dioxygenase 1 (IDO1) enzyme, whose expression is shared in immune regulatory and tumor cells, have been revealed to enhance the anti-tumor immune response. However, poor bioavailability and pharmacokinetics, off-target effects and hurdles in achieving therapeutic drug concentrations at the target tissue often limit the effectiveness of combination therapies.Methods: This work describes the development of novel biomimetic and carrier-free nanobinders (NBs) loaded with both paclitaxel and the IDO1 inhibitor NLG919 in the form of bioresponsive and biomimetic prodrugs. A fine tuning of the preparation conditions allowed to identify NB@5 as the most suitable nanoformulation in terms of reproducibility, stability and in vitro effectiveness.Results and discussion: Our data show that NB@5 effectively binds to HSA in cell-free experiments, demonstrating its protective role in the controlled release of drugs and suggesting the potential to exploit the protein as the endogenous vehicle for targeted delivery to the tumor site. Our study successfully proves that the drugs encapsulated within the NBs are preferentially released under the altered redox conditions commonly found in the tumor microenvironment, thereby inducing cell death, promoting ICD, and inhibiting IDO1.

Multitarget compounds have emerged as promising drug candidates to cope with complex multifactorial diseases, like Alzheimer’s disease (AD). Most multitarget compounds are designed by linking two pharmacophores through a tether chain (linked hybrids), which results in rather large molecules that are particularly useful to hit targets with large binding cavities, but at the expense of suffering from suboptimal physicochemical/pharmacokinetic properties. Molecular size reduction by removal of superfluous structural elements while retaining the key pharmacophoric motifs may represent a compromise solution to achieve both multitargeting and favorable physicochemical/PK properties. Here, we report the stepwise structural simplification of the dihydroxyanthraquinone moiety of a rhein–huprine hybrid lead by hydroxy group removal—ring contraction—ring opening—ring removal, which has led to new analogs that retain or surpass the potency of the lead on its multiple AD targets while exhibiting more favorable drug metabolism and pharmacokinetic (DMPK) properties and safety profile. In particular, the most simplified acetophenone analog displays dual nanomolar inhibition of human acetylcholinesterase and butyrylcholinesterase (IC50 = 6 nM and 13 nM, respectively), moderately potent inhibition of human BACE-1 (48% inhibition at 15 µM) and Aβ42 and tau aggregation (73% and 68% inhibition, respectively, at 10 µM), favorable in vitro brain permeation, higher aqueous solubility (18 µM) and plasma stability (100/96/86% remaining in human/mouse/rat plasma after 6 h incubation), and lower acute toxicity in a model organism (zebrafish embryos; LC50 >> 100 µM) than the initial lead, thereby confirming the successful lead optimization by structural simplification.

Diabetic kidney disease (DKD) is a major cause of morbidity and mortality in individuals with type 2 diabetes mellitus (T2DM). The aim of this study was to investigate whether albumin structural alterations correlate with DKD severity and evaluate whether native and reduced albumin concentrations could complement the diagnosis of DKD. To this end, one hundred and seventeen T2DM patients without (n = 42) and with (n = 75) DKD (DKD I-III upon KDIGO classification) were evaluated; the total albumin concentration (tHA) was quantified by a bromocresol green assay, while structural alterations were profiled via liquid chromatography–high-resolution mass spectrometry (LC-HRMS). The concentrations of native albumin (eHA, effective albumin) and reduced albumin (rHA) were subsequently assessed. The HRMS analyses revealed a reduced relative amount of native albumin in DKD patients along with an increased abundance of altered forms, especially those bearing oxidative modifications. Accordingly, both eHA and rHA values varied during the stages of progressive renal failure, and these alterations were dose-dependently correlated with renal dysfunction. A ROC curve analysis revealed a significantly greater sensitivity and specificity of eHA and rHA than of tHA for diagnosing DKD. Importantly, according to the multivariate logistic regression analysis, the eHA was identified as an independent predictor of DKD.

Multi-target drug discovery is one of the most active fields in the search for new drugs against Alzheimer's disease (AD). This is because the complexity of AD pathological network might be adequately tackled by multi-target-directed ligands (MTDLs) aimed at modulating simultaneously multiple targets of such a network. In a continuation of our efforts to develop MTDLs for AD, we have been focusing on the molecular hybridization of the acetylcholinesterase inhibitor tacrine with the aim of expanding its anti-AD profile. Herein, we manipulated the structure of a previously developed tacrine-quinone hybrid (1). We designed and synthesized a novel set of MTDLs (2-6) by replacing the naphthoquinone scaffold of 1 with that of 2,5,8-quinolinetrione. The most interesting hybrid 3 inhibited cholinesterase enzymes at nanomolar concentrations. In addition, 3 exerted antioxidant effects in menadione-induced oxidative stress of SH-SY5Y cells. Importantly, 3 also showed low hepatotoxicity and good anti-amyloid aggregation properties. Remarkably, we uncovered the potential of the quinolinetrione scaffold, as a novel anti-amyloid aggregation and antioxidant motif to be used in further anti-AD MTDL drug discovery endeavors.

m this paper, the dev'cpment of efficient enanticsetective S1PLC methois for the dnalysis of Rile nenaon iota ran substituted phenethylamines, two sukstituted tryptan >> nes, and three suustituted cath:nones is described. For the first time, reversed phase (eluents made up ..ith acidic water methanol snlutions) and polar-ionic (eluent made up with an acetoniuile methanol solution incmporatinj both an acidic and a basic additive;i conditions fully compatible with mass specunmen-y (MS) detectors were applied with a chiral stationary phase (CEP) incmporating the (+)-(18-crovm-6)-tetracarboxylic acid chiral selector. Enantioresolution was achieved for nine cornpounds with a and R5 factors up to 1.32 and 5.12, respectively. Circular diclunism (CD) detection, CD spectroscopy in stopped-flow mode and quantdm mechanical (QM) calculations were successfidly employed to investigate the absolute stereochemisny of mephedrone, methylone and butylone and allowed to establish a (R)<(S) enantiomeric elution order for these compounds on the chosen CSP. Whole blood miniaturized samples collected by means of volurnetric absorptive microsampling (VAMS) technoloy and fortified with the target analytes were extracted following an optirnized protocol and effectively analysed by means of an ultra-high performance liquid chrornatography-MS system. By this way a proof-ofconcept procedtun was applied, demonstrating the suitability of the method for quali-quantitative enantioselective assessment of the selected psychoactive substances in advanced biological microsamples. VAMS microsamplers including a polypropylene handle topped with a small tip of a polymeric porous material were used and allowed to voltunetrically collect small aliquots of whole blood (10 gL) independently from its density. Highly appreciable volumetric acctuacy (bias, in the -8.7-8.1% range) and precision (% CV, in the 2.8-5.9% range) noned out.

Abstract Background Exopolysaccharides (EPS) secreted by beneficial lactobacilli exert a plethora of positive activities, but little is known about their effects on biofilms of opportunistic vaginal pathogens and especially on biofilms of lactobacilli themselves. Here, the EPS produced by six vaginal lactobacilli, belonging to Lactobacillus crispatus (BC1, BC4, BC5) and Lactobacillus gasseri (BC9, BC12, BC14) species were isolated from cultural supernatants and lyophilized. Results Lactobacillus EPS were chemically characterized in terms of monosaccharide composition by liquid chromatography (LC) analysis coupled to UV and mass spectrometry (MS) detection. Moreover, the ability of EPS (0.1, 0.5, 1 mg/mL) to stimulate the biofilm formation of lactobacilli and to inhibit the formation of pathogens’ biofilms was evaluated by crystal violet (CV) staining and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Isolated EPS (yields 133–426 mg/L) were heteropolysaccharides mainly composed of d-mannose (40–52%) and d-glucose (11–30%). For the first time we demonstrated that Lactobacillus EPS were able to stimulate in a dose-dependent manner (p < 0.05) the formation of biofilms of ten strains belonging to L. crispatus, L. gasseri and Limosilactobacillus vaginalis species, in terms of cell viability (84–282% increase at 1 mg/mL) and especially biofilm biomass (40–195% increase at 1 mg/mL), quantified with MTT assay and CV staining, respectively. EPS released from L. crispatus and L. gasseri were found to better stimulate the biofilms of the same producer species rather than that of other species, including producing strains themselves and other strains. Conversely, the biofilm formation of bacterial (Escherichia coli, Staphylococcus spp., Enterococcus spp. and Streptococcus agalactiae) and fungal (Candida spp.) pathogens was inhibited. The anti-biofilm activity was dose-dependent and was more marked for L. gasseri-derived EPS (inhibition up to 86%, 70%, and 58% at 1 mg/mL, 0.5 mg/mL, and 0.1 mg/mL, respectively), whilst L. crispatus-derived EPS resulted overall less efficient (inhibition up to 58% at 1 mg/mL and 40% at 0.5 mg/mL) (p < 0.05). Conclusions Lactobacilli-derived EPS favour the biofilm formation of lactobacilli preventing, at the same time, that of opportunistic pathogens. These results support the possible employment of EPS as postbiotics in medicine as a therapeutic/preventive strategy to counteract vaginal infections.

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer still lacking effective treatment options. Chemotherapy in combination with immunotherapy can restrict tumor progression and repolarize the tumor microenvironment towards an anti-tumor milieu, improving clinical outcome in TNBC patients. The chemotherapeutic drug paclitaxel had been shown to induce immunogenic cell death (ICD), whereas inhibitors of the indoleamine 2,3-dioxygenase 1 (IDO1), whose expression is shared in immune regulatory and tumor cells, have been revealed to enhance the anti-tumor immune response. However, poor bioavailability and pharmacokinetic, off-target effects and hurdles in achieving therapeutic drug concentrations at the target tissue often limit the effectiveness of combination therapies. This work describes the development of novel biomimetic and carrier-free nanobinders (NB) loaded with both paclitaxel and the IDO1 inhibitor NLG919 in the form of bioresponsive prodrugs, and capable of hijacking human serum albumin (HSA). A fine tuning of the preparation conditions allowed to identify NB@5 as the best-performing prodrugs-based nanoformulation. Our data show that NB@5 effectively binds with HSA, demonstrating its protective role in the controlled release of drugs in vitro and suggesting that NB could exploit the protein as the endogenous vehicle for targeted delivery to the tumor site. Our study successfully demonstrates that the drugs encapsulated within the nanobinders are preferentially released under the altered redox conditions commonly found in the tumor microenvironment, thereby inducing cell death, promoting ICD, and inhibiting IDO1. This study highlights the potential of prodrugs-based nanobinders as a promising avenue for the targeted chemoimmunotherapy of TNBC.