Triple-negative breast cancer (TNBC) remains a major clinical challenge due to its aggressive behavior, high metastatic potential, and limited therapeutic options, as current treatments often constrained by resistance an...Triple-negative breast cancer (TNBC) remains a major clinical challenge due to its aggressive behavior, high metastatic potential, and limited therapeutic options, as current treatments often constrained by resistance and toxicity. Here, ten structurally distinct macrocyclic alkaloids were discovered from sp. Compounds and possessed the 13-membered macrocyclic ring alongside the novel spiro junction at C-3/C-18. Among them, phomapyrrolidone A (Ppd A) was the most active compound in the series, showing the lowest IC values and favorable selectivity toward TNBC cells. Integrated proteomic profiling, bioinformatic analysis, and clinical tissue validation converged on nucleolin (NCL) as a direct and functionally relevant target. Mechanistically, Ppd A binds to the Gln558 residue of NCL, disrupts the NCL-EGFR complex, and suppresses EGFR-PI3K-AKT signaling, leading to apoptosis and tumor growth inhibition. These findings establish NCL as a promising therapeutic target and highlight hirsutellone-type alkaloids as a novel class of NCL modulators for TNBC treatment.
Metallodrugs with mechanisms distinct from cisplatin are promising for overcoming drug resistance and enhancing anticancer efficacy. In this study, a bipyridine-based ligand and its Ni(II) complex were synthesized and ch...Metallodrugs with mechanisms distinct from cisplatin are promising for overcoming drug resistance and enhancing anticancer efficacy. In this study, a bipyridine-based ligand and its Ni(II) complex were synthesized and characterized using elemental analysis, nuclear magnetic resonance (NMR), matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), ultraviolet-visible (UV-vis), Fourier transform infrared (FT-IR), and electrochemical techniques. Density functional theory (DFT) calculations at the B3LYP/6-31G(d,p) level supported the molecular structure. Interaction with calf thymus DNA and bovine serum albumin was confirmed through spectroscopic and biophysical methods, supported by molecular docking studies. The Ni(II) complex exhibited significant cytotoxicity against MG-63 and HCT-116 cancer cells under both dark and near-infrared irradiation, with low toxicity toward normal HEK293 cells. It showed higher activity than the free ligand, along with notable antioxidant and antimicrobial properties. Additionally, efficient photocatalytic degradation of Eosin Y (up to 93% under irradiation) highlights its multifunctional potential in biomedical and environmental applications.
DNA-encoded libraries (DELs) enable rapid discovery of large pools of small organic ligands against target proteins. Currently available hit validation methodologies are limited in their ability to characterize binding k...DNA-encoded libraries (DELs) enable rapid discovery of large pools of small organic ligands against target proteins. Currently available hit validation methodologies are limited in their ability to characterize binding kinetics for a large number of molecules. Only a subset of hits identified by DEL screening are followed up on, while many potentially relevant binders remain uncharacterized. Here, we propose focal molography as a high-throughput, label-free optical methodology for parallel kinetic measurements of DEL-derived hits. The methodology was applied to measure binding kinetics ( and ) and dissociation constants () of DEL-derived ligands against carbonic anhydrase IX and of known ligands to fibroblast activation protein. The binding parameters obtained were consistent with fluorescence polarization, surface plasmon resonance, and inhibition measurements. The data reported in this manuscript support the use of focal molography as a robust DEL-compatible technology for quantitative, high-throughput hit validation.
New half-sandwich iridium(III) compounds [Ir(η-Cp)Cl(L1-3)]PF (-), combining Cp* or Cp with N,P-coordinated phosphinoalkylamines L1-L3, were tested in different cancer cells (2D and 3D cultures), including MOR/CPR cispla...New half-sandwich iridium(III) compounds [Ir(η-Cp)Cl(L1-3)]PF (-), combining Cp* or Cp with N,P-coordinated phosphinoalkylamines L1-L3, were tested in different cancer cells (2D and 3D cultures), including MOR/CPR cisplatin-resistant lung carcinoma. Best-performing compound outperformed its Cp analogue and cisplatin in MOR/CPR cells while sparing noncancerous cells. Multiomics profiling shows a non-DNA-targeted mechanism: rapid integrated stress response with ER stress (DDIT3/CHOP) and oxidative stress (HMOX1, ATF3), nucleolar stress, and primary inhibition of ribosome biogenesis and mitochondrial translation. These changes drive translational shutdown, suppression of oxidative phosphorylation with a glycolytic shift, and G arrest, alongside endolysosomal remodeling (enhanced vesicular uptake, reduced degradative capacity) that favors intracellular retention. The phenotype is predominantly cytostatic with apoptotic priming. In vivo, suppressed tumor growth and activated apoptosis with low systemic toxicity. Compound thus emerges as a promising prototype Ir(III) metallodrug that disrupts nucleolar, mitochondrial, and lysosomal homeostasis to overcome resistance.
Microtubules are critical regulators of mitotic progression and cancer cell proliferation. Herein, we report a novel 2-amino-3-cyanopyridine-based scaffold that functions as a dual inhibitor of tubulin polymerization and...Microtubules are critical regulators of mitotic progression and cancer cell proliferation. Herein, we report a novel 2-amino-3-cyanopyridine-based scaffold that functions as a dual inhibitor of tubulin polymerization and cyclin G-associated kinase (GAK). Structure-activity relationship (SAR) optimization identified KMG-732 with nanomolar to submicromolar cytotoxicity across diverse cancer cell lines. Mechanistic investigations revealed that KMG-732 targets the colchicine-binding site, effectively disrupting microtubule assembly and triggering mitotic arrest followed by apoptosis. Beyond its antimitotic effects, KMG-732 significantly attenuates cancer cell migration and invasion. Importantly, KMG-732 displayed favorable pharmacokinetic properties and showed minimal P-glycoprotein-mediated efflux. KMG-732 demonstrated robust antitumor efficacy in both organoids and xenograft models, maintaining a superior safety profile with reduced systemic toxicity compared to colchicine. Kinome profiling and biochemical assays confirmed selective GAK inhibition, distinguishing KMG-732 from traditional tubulin inhibitors. Collectively, KMG-732 represents a promising dual-targeting antimitotic lead candidate with enhanced efficacy and therapeutic window for cancer treatment.
Poddar SK, Yang Y, Pal P
… +13 more, Jin Z, Pei J, Xiao Y, Hu W, Zhang P, Adekunbi P, Thummuri D, Yuan Y, Lv D, Pelletier JPR, Zhang W, Zhou D, Zheng G
Glaucoma is a leading cause of irreversible blindness worldwide, and lowering intraocular pressure (IOP) remains the only proven method to delay progression. While topical carbonic anhydrase (CA) inhibitors are establish...Glaucoma is a leading cause of irreversible blindness worldwide, and lowering intraocular pressure (IOP) remains the only proven method to delay progression. While topical carbonic anhydrase (CA) inhibitors are established treatments, more potent and selective agents are needed. We report a series of subnanomolar and highly selective human carbonic anhydrase (hCA) II inhibitors developed through a multistage fragment growth strategy. By incrementally expanding a weakly bound sulfanilamide anchor to precisely complement the enzyme's topology, we achieved a remarkable 15,000-fold increase in potency, resulting in subnanomolar inhibition and exceptional selectivity. High-resolution X-ray crystallography revealed a distinct L-shaped configuration, effectively occupying the entire active site and establishing unique interactions with a peripheral α-helix. In rabbit glaucoma models, these inhibitors demonstrated superior IOP-lowering activity. Furthermore, the inhibitors exhibited exceptional metabolic stability, minimal ocular toxicity, and favorable safety profiles. These results demonstrate multistage fragment growth as a robust strategy for designing next-generation glaucoma therapeutics.
The SARS-CoV-2 Spike receptor-binding domain (S-RBD)/hACE2 interaction represents a challenging protein-protein interaction (PPI) target due to its large, shallow binding interface. Here, in silico alanine mutagenesis gu...The SARS-CoV-2 Spike receptor-binding domain (S-RBD)/hACE2 interaction represents a challenging protein-protein interaction (PPI) target due to its large, shallow binding interface. Here, in silico alanine mutagenesis guided the structure-based design of constrained peptidomimetics that reproduce key hACE2 recognition elements. α1-helix-derived mimetics (Glu23-Ser44) were stabilized using peptide stapling strategies, while antiparallel β-sheet mimetics (Thr347-Leu359) were generated through head-to-tail macrocyclization incorporating a d-Pro/l-Pro motif. Covalent linkage of these two secondary structure mimetics yielded proteomimetic , designed to preserve the spatial organization of the hACE2 binding interface. Compound selectively binds SARS-CoV-2 S-RBD and disrupts the S-RBD/hACE2 interaction in biophysical and cellular assays, inhibiting pseudovirus entry (IC of 6.6 μM). Importantly, displays high stability in lung epithelial models ( > 24 h) and low epithelial permeability ( = 2.03 × 10 cm·s), supporting its potential for intranasal antiviral delivery. These findings establish a proof-of-concept for proteomimetics as promising inhibitors of challenging PPIs.
CD1d-restricted glycolipids have emerged as a cornerstone in the development of next-generation immunotherapies. This perspective provides a comprehensive update on structure-activity relationships, specifically examinin...CD1d-restricted glycolipids have emerged as a cornerstone in the development of next-generation immunotherapies. This perspective provides a comprehensive update on structure-activity relationships, specifically examining how modifications of the prototypical CD1d ligand α-galactosylceramide (α-GalCer) to the galactosyl headgroup, phytosphingosine base, and fatty acyl chain dictate the modulation of invariant natural killer T (iNKT) cell responses. We explore the key pharmacophores and receptor interactions that polarize subsequent immune response toward either a pro-inflammatory Th1 or an anti-inflammatory Th2 phenotype. Building upon the structural evolution of sophisticated chemotypes, we evaluate the potential of these agonists in synergistic combinations with other adjuvants. Furthermore, we highlight the emerging transition toward fully synthetic self-adjuvanting vaccines, which ensure cellular colocalization and coordinated activation by covalently integrating antigens with glycolipid agonists. Collectively, these advancements underscore the transformative potential of tailored glycolipid design in engineering specific and durable immunity against cancer and infectious diseases.
Pyruvate carboxylase (PC) replenishes tricarboxylic acid cycle intermediates, driving cancer metabolic reprogramming. To improve the metabolic stability of erianin, a potent PC inhibitor from Lindl, we designed and synt...Pyruvate carboxylase (PC) replenishes tricarboxylic acid cycle intermediates, driving cancer metabolic reprogramming. To improve the metabolic stability of erianin, a potent PC inhibitor from Lindl, we designed and synthesized 55 derivatives, culminating in the identification of which exhibited potent PC inhibition (IC = 1.74 nM) and suppressed HCC cell viability (IC = 25.18 nM), comparable to erianin. Notably, demonstrated significantly improved stability, with a half-life ( = 1.21 h) much longer than erianin ( ∼ 0.1 h). Mechanistically, suppressed HCC proliferation and metastasis by inducing apoptosis and ferroptosis. Moreover, it promoted mitochondrial oxidative stress and inhibited glycolysis, thereby sensitizing cells to glutamine deprivation. , exhibited comparable antitumor efficacy but improved safety compared to sorafenib. With its potent PC inhibition and favorable drug-like properties, represented a promising therapeutic candidate for HCC treatment.
Proteolysis-targeting chimeras (PROTACs) are attractive targeted protein degradation modalities, while their clinical advancement is limited by high molecular weight, suboptimal bioavailability, low cellular permeability...Proteolysis-targeting chimeras (PROTACs) are attractive targeted protein degradation modalities, while their clinical advancement is limited by high molecular weight, suboptimal bioavailability, low cellular permeability, and unsatisfactory in vivo efficacy. Herein, we developed a self-assembled nanoparticle system (PRO NPs) constructed from amphiphilic PROTACs. Owing to their intrinsic amphiphilicity, PRO NPs can spontaneously assemble into stable nanoparticles without external excipients, simplifying fabrication and alleviating carrier-associated toxicity. Compared with the free EGFR-targeting PROTAC, PRO NPs exhibited enhanced aqueous solubility and efficient EGFR degradation in HCC827 cells. In vivo, PRO NPs functionalized with hyaluronic acid (HA) through electrostatic adsorption could actively accumulate at tumor sites via CD44-mediated targeting, exerting potent antitumor efficacy (TGI = 76.8%) with reduced systemic toxicity. Collectively, these findings provide novel insights into nano-PROTAC design and a promising strategy for the development of targeted protein degradation therapies.
Myeloid differentiation factor 88 (MyD88) is a critical mediator of inflammatory signaling. However, the development of MyD88 inhibitors has been limited, and there are no reports on MyD88 degraders. Based on our previou...Myeloid differentiation factor 88 (MyD88) is a critical mediator of inflammatory signaling. However, the development of MyD88 inhibitors has been limited, and there are no reports on MyD88 degraders. Based on our previously identified MyD88 inhibitor and employing a template-assisted modification strategy, we designed and synthesized 40 novel derivatives, and identified the optimal compound , which could inhibit nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway activation by facilitating the interaction between ring finger protein 126 (RNF126) (an E3 ubiquitin ligase) and MyD88, thereby promoting ubiquitination and subsequent degradation of MyD88. The subacute toxicity test indicated that had good safety profile. Furthermore, it demonstrated significant anti-inflammatory effects in both cecal ligation and puncture (CLP) and lipopolysaccharide (LPS) induced acute lung injury (ALI) models, effectively ameliorating ALI symptoms. These findings suggest that holds considerable promise as a MyD88-targeting molecular glue degrader for the treatment of ALI.
Diamanti E, Cremers AJH, Li Y
… +16 more, Exapicheidou I, Baumann C, Gibson P, Shams A, Martin L, Becker R, Setyawati I, Zeimetz L, Haupenthal J, Witschel M, Slotboom DJ, Herrmann J, Rox K, Hamed MM, Veening JW, Hirsch AKH
Energy-Coupling Factor (ECF) transporters are a hitherto underexplored target involved in the uptake of several micronutrients in bacteria and are absent in humans. Here, in , we demonstrate that the genes encoding ECF t...Energy-Coupling Factor (ECF) transporters are a hitherto underexplored target involved in the uptake of several micronutrients in bacteria and are absent in humans. Here, in , we demonstrate that the genes encoding ECF transporters are highly conserved and their expression is crucial for causing bacterial infection in both murine () and human () infection models. Next, we demonstrated that the antimicrobial activity of our inhibitors against is related to the level of the ECF transporter expressed by the bacterium, confirming the target engagement of this chemical class in . The pharmacokinetic studies conducted revealed high peroral bioavailability for , which was also assessed in a murine neutropenic lung-infection model with , and observed a one log reduction in bacterial load compared to vehicle. This work sets the stage for an innovative approach to combat -derived infections by targeting the ECF transporters with the novel chemotype reported.
Protein oligomerization is functionally essential for many enzymes, yet small-molecule strategies that directly target dimer interfaces remain challenging. Cysteine residues at protein dimer interfaces offer chemically a...Protein oligomerization is functionally essential for many enzymes, yet small-molecule strategies that directly target dimer interfaces remain challenging. Cysteine residues at protein dimer interfaces offer chemically addressable sites for modulating oligomeric assembly. Here, we used covalent DNA-encoded chemical library (CoDEL) screening to identify site-selective covalent hits targeting the PHGDH dimer interface. Covalent hits emerging from CoDEL screening were optimized to yield a selective covalent inhibitor engaging the interfacial Cys281. A representative compound, , potently disrupts PHGDH dimerization and inhibits its enzymatic activity, restores sensitivity to EGFR tyrosine kinase inhibitors in resistant lung adenocarcinoma cells in vitro, and exhibits antitumor efficacy in mouse xenograft models. Together, these findings establish dimer-interface cysteine targeting as a mechanism-based and therapeutically relevant strategy for modulating PHGDH function, and highlight the potential of CoDEL for discovering covalent inhibitors of protein-protein interfaces.
Puxeddu M, Cui Z, Colla C
… +19 more, Nalli M, Manetto S, Ciogli A, Cuřínová P, Bufano M, Toto A, Gianni S, Pastore A, Stornaiuolo M, Baldini E, Ulisse S, Kopecka J, Riganti C, Bigogno C, Dondio G, Liu T, Coluccia A, Regina G, Silvestri R
Dishevelled (DVL) proteins are key mediators of the Wnt/β-catenin signaling pathway, involved in signal transduction from membrane receptors to intracellular effectors. DVL up-regulation has often been correlated with tu...Dishevelled (DVL) proteins are key mediators of the Wnt/β-catenin signaling pathway, involved in signal transduction from membrane receptors to intracellular effectors. DVL up-regulation has often been correlated with tumor progression and metastasis. DVL1 was found overexpressed in multidrug-resistant colorectal cancer (CRC) cells. Here, we describe the synthesis of new indole-2-carboxamides as DVL1 inhibitors. Compound (,)- showed potent DVL1 inhibition with IC of 0.97 ± 0.21 μM and specific binding to the DVL1 PDZ domain. (,)- strongly reduced β-catenin expression in HCT116 CRC cells and significantly decreased tumor volume and weight in a xenograft model. Additionally, (,)- inhibited P-glycoprotein (P-gp), restoring sensitivity to doxorubicin (DOX) in HT29/DX CRC chemoresistant cells. (,)- demonstrated high metabolic stability in human liver microsomes, and an acceptable pharmacokinetic profile following IV administration in mice. Our findings indicate that compound (,)- represents a promising dual-targeting antitumor candidate for CRC treatment.
Timari MP, Paczal A, Herner A
… +26 more, Molnar M, Madarasz Z, Nyerges M, Bedford ST, Brooks T, Davidson J, Daniels Z, Dodsworth M, Dokurno P, Murray JB, Parsons R, Sanders E, Smith J, Webb P, Whitehead N, Hubbard RE, Starck JB, Maragno AL, Le Toumelin-Braizat G, Bresson L, Rocchetti F, Demarles D, Colland F, Geneste O, Kotschy A, Novak T
Evasion of apoptosis is a hallmark of cancer. Deregulation of BCL-XL, a member of the BCL-2 family of proteins, has been linked to the development of various tumor types. This study presents the design and synthesis of B...Evasion of apoptosis is a hallmark of cancer. Deregulation of BCL-XL, a member of the BCL-2 family of proteins, has been linked to the development of various tumor types. This study presents the design and synthesis of BCL-XL inhibitors with novel mono- and bicyclic cores. The new structural features were optimized to combine high binding efficiency with the opening of diverse novel vectors for additional modifications. The lead compounds exhibited picomolar affinities and significant cellular potency in the BCL-XL-dependent MOLT-4 cell line, which also translated into marked tumor growth inhibition in a xenograft study. These findings highlight the potential of BCL-XL inhibitors as therapeutic agents in cancer treatment by targeting the apoptotic intrinsic pathway.
To realize spatiotemporal control of nitric oxide (NO), a key signaling molecule for vasodilation in biological systems, we previously developed photocontrollable NO releasers such as NORD-1, whose NO release is triggere...To realize spatiotemporal control of nitric oxide (NO), a key signaling molecule for vasodilation in biological systems, we previously developed photocontrollable NO releasers such as NORD-1, whose NO release is triggered by photoinduced electron transfer (PeT) to a light-harvesting moiety. Here, we aimed to improve the PeT-driven NO release by investigating the structure-activity relationship of the rhodamine antenna, focusing on modifications of amino groups at the 3-/6-positions and the element at the 10-position. NO release from selenium- or tellurium-containing NO releasers (NO-SeR and NO-TeR) was significantly enhanced, whereas a phospharhodamine-type NO releaser (NO-POR) exhibited no detectable photodecomposition. Quantum-chemical calculations suggested that the charge-transfer state for NO-POR is stabilized compared to other NO releasers. We confirmed that NO-TeR showed more efficient vasodilation than NORD-1 in studies. These findings are expected to facilitate the design of new PeT-driven compounds for biological and therapeutic applications.
This work presents a novel, promising, and efficient strategy to develop potential Pt(IV) prodrugs for cancer chemotherapy while to simultaneously overcome the ″undruggability″ of CDC25A. New series of Pt(IV) complexes (...This work presents a novel, promising, and efficient strategy to develop potential Pt(IV) prodrugs for cancer chemotherapy while to simultaneously overcome the ″undruggability″ of CDC25A. New series of Pt(IV) complexes (-) bearing 1,4-naphthoquinone (NQ, a natural active skeleton) derivatives (-) as potential dual CDC25A/NF-κB inhibitory ligands were synthesized and characterized. Their in vitro and in vivo anticancer activities were subsequently evaluated. Mechanistic investigations revealed that , the most potent candidate with ligand , effectively depleted CDC25A levels in A2780 cancer cells through a synergistic effect mediated by DNA damage. Thus, triggered multimodal anticancer mechanisms, which included reactive oxygen species/endoplasmic reticulum stress-mediated mitochondrial apoptotic pathway, DNA damage coupled with S-phase cell cycle arrest, and autophagy-dependent ferroptosis. In A2780 xenograft models, administrated with a dosage of 8 mg/kg manifested superior tumor growth inhibition than both cisplatin and "cisplatin + " combination, along with satisfying low toxicity.
G2A inhibition has recently been proposed as a novel therapeutic approach to treat oxaliplatin-induced neuropathic pain (OINP) and breast cancer. However, very few G2A antagonists are known to date. In this study, we rep...G2A inhibition has recently been proposed as a novel therapeutic approach to treat oxaliplatin-induced neuropathic pain (OINP) and breast cancer. However, very few G2A antagonists are known to date. In this study, we report the discovery of a novel series of G2A antagonists developed within our research group, along with the first comprehensive structure-activity relationship (SAR) investigation for this class of compounds. Utilizing a rational design approach, we systematically explored the effects of structural modifications on G2A receptor binding and functional activity. The SAR study identified key molecular features critical for potent G2A inhibition. Two of the newly discovered compounds exhibited submicromolar activity and acceptable selectivity profile among GPCRs.
Cancer immunotherapy harnesses the host immune system to treat tumors. TLR-9, a key bridge between innate and adaptive immunity, is activated by CpG oligodeoxynucleotides (ODNs). However, linear CpG ODNs suffer from rapi...Cancer immunotherapy harnesses the host immune system to treat tumors. TLR-9, a key bridge between innate and adaptive immunity, is activated by CpG oligodeoxynucleotides (ODNs). However, linear CpG ODNs suffer from rapid nuclease degradation , resulting in a short half-life and reduced immunostimulatory activity. Here, we developed covalently closed circular CpG ODNs based on FDA-approved sequences (1018 and 7909). The circular CpG ODNs exhibited superior nuclease resistance and significantly enhanced immunostimulatory activity in both human and murine immune cells. In the poorly immunogenic B16-F10 melanoma model, combining circular CpG ODNs with dacarbazine (DTIC) chemotherapy produced markedly stronger and more sustained antitumor efficacy, with enhanced tumor regression and improved survival, compared to linear CpG ODN/DTIC combinations. Our findings establish circular CpG ODNs as a promising platform for cancer immunotherapy, offering improved long-term stability and potent immunomodulatory capacity, particularly in combination with chemotherapy.