The activation of thyroid hormone receptor β (THRβ) represents a highly validated therapeutic strategy for metabolic dysfunction-associated steatohepatitis (MASH). Nevertheless, attaining high selectivity for THRβ is cru...The activation of thyroid hormone receptor β (THRβ) represents a highly validated therapeutic strategy for metabolic dysfunction-associated steatohepatitis (MASH). Nevertheless, attaining high selectivity for THRβ is crucial to prevent the unintended activation of the cardiotoxic THRα subtype. In this work, we capitalized on the flexibility of the hydrophobic binding pocket of THRβ and synthesized a series of cyclized indole-carboxylic acid derivatives using a conformational restriction strategy. The representative compound exhibited potent THRβ agonism (EC = 36 nM) and no discernible activity against THRα, confirming its high functional selectivity. In vitro druggability assessments revealed that possesses excellent metabolic stability across species and favorable oral pharmacokinetics in mice. In a murine MASH model, significantly ameliorated hepatic steatosis, serum biomarkers, and liver gene expression profiles, without observable cardiac toxicity. Overall, these findings suggest that is a promising candidate for the development of selective THRβ-targeted therapies for MASH.
Nitroreductase (NTR), a hypoxia-upregulated enzyme, is a promising biomarker for solid tumors, driving demand for effective biosensors. Cyanine dyes are prominent fluorophores widely used in fluorescence imaging, tumor p...Nitroreductase (NTR), a hypoxia-upregulated enzyme, is a promising biomarker for solid tumors, driving demand for effective biosensors. Cyanine dyes are prominent fluorophores widely used in fluorescence imaging, tumor phototheranostics, and image-guided surgery. Guided by atomic engineering through strategic heteroatom substitution (C, O, and S), we designed four NTR-activated probes. Optical characterization identified as the lead candidate, exhibiting near-infrared emission (λ = 712 nm) and over 18-fold fluorescence enhancement upon activation. The probe enabled precise imaging of hypoxic NTR and distinguished malignant from nonmalignant cells, while and studies supported the use of the probe in intraoperative tumor navigation. Additionally, using as a screening tool, we identified luteolin as a potential NTR modulator that exhibited concentration-dependent suppression of NTR-associated fluorescence activation and significant antitumor activity . Collectively, this continuum─from atomic-scale molecular design to novel NTR inhibitor discovery─provides an innovative model for drug screening.
Acetaminophen (APAP) overdose is a leading cause of acute liver failure, and -acetylcysteine therapy is highly time dependent. APAP hepatotoxicity results from cytochrome P450-mediated oxidative bioactivation to -acetyl-...Acetaminophen (APAP) overdose is a leading cause of acute liver failure, and -acetylcysteine therapy is highly time dependent. APAP hepatotoxicity results from cytochrome P450-mediated oxidative bioactivation to -acetyl--benzoquinone imine (NAPQI). Here, we report an electron effect-guided metabolic reprogramming strategy based on amide-to-thioamide modification to suppress APAP bioactivation while preserving efficacy. We synthesized a thioamide-modified APAP analogue, SAPAP, and evaluated its metabolism, hepatic safety, and pharmacological activity. SAPAP redirected metabolic flux toward glucuronide and sulfate conjugation while minimizing oxidative metabolite formation. In acute overdose and 28-day subchronic dosing models, SAPAP caused markedly less liver injury than equimolar APAP, with improved histopathology, reduced serum transaminases, and attenuated inflammatory responses. SAPAP also retained analgesic and antipyretic efficacy in established models. These findings demonstrate that thiocarbonyl-driven electronic effects can decouple therapeutic efficacy from hepatotoxic liability, offering a preventive strategy against APAP-induced liver injury.
Łęcka M, Gorzeń O, Ćwilichowska-Puślecka N
… +11 more, Nguyen J, Majchrzak M, Pippa V, Jakimowicz P, Wiśniewski J, Dołęga-Kozierowski B, Kasprzak P, Turk B, Drąg M, Matkowski R, Poręba M
Cysteine cathepsins, particularly cathepsin S, regulate proteolytic signaling in cancer progression and immune modulation, yet selective tools for individual cathepsins remain limited. Here, we report the design of cathe...Cysteine cathepsins, particularly cathepsin S, regulate proteolytic signaling in cancer progression and immune modulation, yet selective tools for individual cathepsins remain limited. Here, we report the design of cathepsin S-selective probes and cathepsin S-cleavable antibody-drug conjugates (ADCs) using substrate profiling with unnatural amino acids. Hybrid Combinatorial Substrate Library (HyCoSuL) screening identified selective tetrapeptide motifs that were used to develop optimized fluorogenic substrates, irreversible inhibitors, and fluorescent activity-based probes with high selectivity for cathepsin S in biochemical and cellular assays. These peptide motifs were then incorporated as cleavable linkers in MMAE-based ADCs targeting HER-2 or TROP-2, enabling cathepsin S-dependent cytotoxicity across breast cancer models with distinct target-expression profiles. Finally, anti-cathepsin S antibodies combined with CyTOF analysis revealed the spatial distribution of cathepsin S and its coexpression with HER-2 and TROP-2 in breast cancer patient samples, suggesting that cathepsin S profiling may help inform future patient stratification strategies for cathepsin S-activated ADC therapy.
Targeted protein degradation represents an emerging new drug discovery strategy. Herein, we identified a novel and potent dual-target degrader targeting PARP1 and IKZF3 based on our previously reported PARPi Thioparib. ,...Targeted protein degradation represents an emerging new drug discovery strategy. Herein, we identified a novel and potent dual-target degrader targeting PARP1 and IKZF3 based on our previously reported PARPi Thioparib. , compound () exerted impressive antiproliferative activities against HR-deficient cancer cells with IC values ranging from 0.006 nM to 39.41 nM. Concurrently, compound displayed potent cell growth inhibitory activities against HR-proficient and clinically used PARPi-resistant cancer cells, with efficacy significantly superior to that of AZD5305. , compound demonstrated excellent tumor growth inhibition (TGI) in the AZD5305-sensitive/-insensitive xenograft model with TGI values of 133.6% and 71.9%, respectively. Collectively, compound may be a powerful therapeutic agent not only for treating PARPi-sensitive tumors but also PARPi-resistant tumors, even HR-proficient tumors.
Endocrine resistance remains a major obstacle in the effective treatment of hormone-receptor-positive breast cancer. mutations, such as Y537S and D538G, are commonly considered to confer such resistance. It is necessary...Endocrine resistance remains a major obstacle in the effective treatment of hormone-receptor-positive breast cancer. mutations, such as Y537S and D538G, are commonly considered to confer such resistance. It is necessary to develop novel therapeutic agents to overcome endocrine resistance. Herein, we developed drug-conjugated Tam-NHC-gold(I) complexes that can target breast cancer cells by the binding of tamoxifen (Tam) to the G protein-coupled estrogen receptor (GPER) present on cell membranes. The privileged complex, , can significantly downregulate ER, inhibit ER downstream signaling pathways, and induce damage-associated molecular pattern (DAMP)-mediated immunogenic cell death (ICD). Mechanistically, RNA-sequencing analysis revealed that can overcome mutant MCF-7 resistance through the RAMP3/CALCR signaling pathway. Moreover, exhibited potent antiproliferative activity on both wild-type and mutant MCF-7 in xenograft mouse models with low toxicity. This study verified that may offer a new approach for the treatment of endocrine-resistant breast cancer.
Antimicrobial resistance (AMR) has severely compromised the efficacy of conventional antibiotics, posing a major threat to global public health. Antimicrobial peptides (AMPs) emerged as a potential solution to combat mul...Antimicrobial resistance (AMR) has severely compromised the efficacy of conventional antibiotics, posing a major threat to global public health. Antimicrobial peptides (AMPs) emerged as a potential solution to combat multidrug-resistant pathogens. However, the clinical applications of AMPs are hindered by their instability, cytotoxicity, and nonspecificity. Herein, we report a series of sulfonyl-γ-AApeptides helical foldamers, which exhibit broad-spectrum antibacterial activity and a high selectivity index against a panel of Gram-positive and -negative strains, including drug-resistant ones. The lead AApeptide AM10 demonstrated potent activity with rapid bactericidal kinetics, strong antibiofilm effects, minimal resistance development, and exceptional stability. Mechanistic studies revealed that AM10 exerts its effect through membrane perturbation and excessive reactive oxygen species (ROS) accumulation. Additionally, AM10 significantly reduced bacterial burden in a mice MRSA infection model, highlighting the sulfonyl-γ-AApeptide foldamers as a promising platform for the development of next-generation antimicrobial peptidomimetics.
Zhou J, Li Z, Sun X
… +17 more, Zhang X, Shi C, Chi X, Lu D, Lyu W, Xiu S, Yang T, Chen Y, Wang Z, Li Q, Zheng R, Du J, Lin Y, Yan J, Li T, Zhang L, Liu Z
Polo-like kinase 1 (PLK1) is a validated cancer therapeutic target, but ATP-competitive PLK1 inhibitors have been limited by dose-limiting toxicities. Here, we designed covalent inhibitors that specifically engage the no...Polo-like kinase 1 (PLK1) is a validated cancer therapeutic target, but ATP-competitive PLK1 inhibitors have been limited by dose-limiting toxicities. Here, we designed covalent inhibitors that specifically engage the noncatalytic Cys67 residue of the ATP domain. Rational optimization of the warhead and core scaffold led to as a highly potent lead candidate. exhibited low nanomolar enzymatic inhibition and exceptional cellular potency (IC = 0.4 nM in K562 leukemia cells), representing a >150-fold improvement over the reversible reference compound BI2536 (61 nM). also showed favorable overall selectivity against nonrelated kinases. Remarkably, achieved 77.6% oral bioavailability in mice. In an orthotopic K562 leukemia xenograft model, oral administration of significantly suppressed the bioluminescence of leukemia burden and extended survival rates to 100% over 42 days without observable toxicity. These findings underscore the potential of as a safe, orally bioavailable, and highly potent PLK1 inhibitor for preclinical development.
We describe the discovery and characterization of , a potent and brain-penetrant lysophosphatidic acid receptor 1 (LPAR1) antagonist with slow association and dissociation kinetics. SAR studies initiated from a literatur...We describe the discovery and characterization of , a potent and brain-penetrant lysophosphatidic acid receptor 1 (LPAR1) antagonist with slow association and dissociation kinetics. SAR studies initiated from a literature lead () led to the identification of compound that possessed a unique urea scaffold responsible for slow but tight binding to LPAR1. Further optimizations that improved PK and metabolic profiles led to the discovery of . efficiently traversed the BBB in multiple preclinical species and was efficacious in preclinical models of neuroinflammatory disorders. possessed excellent ADME properties and was well-tolerated in 28 day GLP (Good Laboratory Practice) toxicity studies in rats and minipigs at doses up to 1000 mg/kg/day. Based on these results and a comprehensive first-in-human enabling preclinical data package, was advanced into clinical development, including an ongoing trial in subjects with chronic osteoarthritis pain or chronic lower back pain (NCT6810245).
CD73 (ecto-5'-nucleotidase) drives immunosuppressive and tumor progression through both enzymatic adenosine production and nonenzymatic mechanisms, limiting the efficacy of current CD73-targeted therapies, including smal...CD73 (ecto-5'-nucleotidase) drives immunosuppressive and tumor progression through both enzymatic adenosine production and nonenzymatic mechanisms, limiting the efficacy of current CD73-targeted therapies, including small-molecule enzymatic inhibitors and antibodies. Here, we reported NUCC-0227579 (C79), a first-in-class proteolysis-targeting chimera (PROTAC) that degraded CD73 via the VHL E3 ligase-dependent proteasomal and lysosomal pathways. C79 eliminated CD73 at the cell surface and intracellular compartments across multiple human cancer cell lines, abolishing nucleotidase activity and more effectively reversing adenosine-mediated immunosuppression than enzymatic inhibitors. C79 enhanced NF-κB/NFAT signaling, increased IFN-γ and TNF-α production, and promoted human CD8 T cell activation and proliferation. In addition, C79 impaired tumor cell metabolic fitness, proliferation, migration, and adhesion through non-nucleotidase-dependent mechanisms. In humanized NSG mouse models of triple-negative breast cancer, C79 significantly suppressed tumor growth while enhancing antitumor immune responses, highlighting CD73 degradation as a mechanistically distinct strategy to overcome the limitations of existing CD73-targeted therapies.
Triple-negative breast cancer (TNBC) lacks effective molecular targets for imaging. Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is overexpressed in the mesenchymal-like (claudin-low) subtype of TNBC. Here, we...Triple-negative breast cancer (TNBC) lacks effective molecular targets for imaging. Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is overexpressed in the mesenchymal-like (claudin-low) subtype of TNBC. Here, we developed two optimized linear peptide radiotracers, DR1 and DR2, derived from the complementarity-determining regions (CDR) of an anti-ROR1 antibody, conjugated with DOTA and labeled with Ga. Both peptides showed nanomolar binding affinities ( = 93.4 ± 8.2 nM for DR1 and 38.0 ± 4.6 nM for DR2) and favorable target-binding properties. [Ga]Ga-DOTA-DR2 exhibited excellent stability (≥95% radiochemical purity after 4 h in human serum), rapid clearance ( = 0.66 ± 0.11 h), and specific cellular uptake. Micro-PET/CT imaging in MDA-MB-231 xenograft mice revealed rapid tumor visualization, with a peak tumor uptake of 2.75 ± 0.61 %ID/g at 1 h and a tumor-to-muscle ratio of 7.98 at 2 h. Blocking studies confirmed ROR1-specific targeting. Collectively, [Ga]Ga-DOTA-DR2 may represent a promising PET tracer for imaging ROR1-positive mesenchymal-like TNBC, providing a foundation for further optimization.
Targeted protein degradation (TPD) has emerged as a transformative strategy in drug discovery, yet the repertoire of E3 ligase recruiters remains limited. Here, we report the discovery of an aldehyde-anchored PROTAC that...Targeted protein degradation (TPD) has emerged as a transformative strategy in drug discovery, yet the repertoire of E3 ligase recruiters remains limited. Here, we report the discovery of an aldehyde-anchored PROTAC that covalently engages the E3 ligase FBXO22 to induce degradation of the histone methyltransferase NSD2 and CDK12. Competitive electrophile screening identified a phenyl aldehyde warhead as optimal, with SAR studies revealing that degradation is highly sensitive to the steric and electronic environment of the aldehyde moiety. The lead degrader, T9, effectively and selectively induces NSD2 degradation across multiple cancer cell lines. Mechanistic investigations confirmed that degradation is dependent on FBXO22, the ubiquitin-proteasome system, and the neddylation pathway, with mutagenesis identifying Cys326 as the critical residue for covalent engagement. This work establishes a stable covalent ligand for FBXO22, expanding chemical space of PROTAC design by introducing an accessible aldehyde-based E3 ligase ligand with broad potential for protein degradation.
Tumor heterogeneity severely limits the efficacy of single-targeted radiopharmaceuticals. FAP and CAIX are important biomarkers overexpressed in tumors and the tumor microenvironment. We designed a PEG-modified heterodim...Tumor heterogeneity severely limits the efficacy of single-targeted radiopharmaceuticals. FAP and CAIX are important biomarkers overexpressed in tumors and the tumor microenvironment. We designed a PEG-modified heterodimeric radioligand, FC, for dual targeting of FAP and CAIX. FC showed high binding affinity toward FAP (IC = 18.2 ± 0.23 nM). [Ga]Ga-FC exhibited significantly higher tumor uptake than monomeric tracers in FAP- or CAIX-positive tumor models at 120 min postinjection. PEG modification markedly enhanced hydrophilicity, tumor retention, and in vivo stability. Mirco-SPECT imaging showed strong tumor accumulation of [Lu]Lu-FC up to 168 h. A single dose of [Lu]Lu-FC (29.6 MBq) achieved complete tumor suppression in HEK293-FAP tumor-bearing mice with no obvious organ toxicity. [Ga]Ga/[Lu]Lu-FC represents a promising theranostic agent for FAP-positive malignancies.
Targeting both the PD-1/PD-L1 axis and NAMPT represents a promising therapeutic strategy to overcome resistance to the immune checkpoint blockade. Herein, we developed a novel series of 4-arylindolines bearing a pyrido[3...Targeting both the PD-1/PD-L1 axis and NAMPT represents a promising therapeutic strategy to overcome resistance to the immune checkpoint blockade. Herein, we developed a novel series of 4-arylindolines bearing a pyrido[3,2-]pyrimidine scaffold as dual-targeting inhibitors. Among these, compound emerged as the most promising inhibitor, exhibiting IC values of 7.5 and 18.8 nM against the PD-1/PD-L1 interaction and NAMPT, respectively, along with favorable metabolic stability. investigations revealed that significantly enhanced T cell-mediated tumor cell killing, elevated T cell proliferation and CD8 T cell proportion, and interfered with intracellular NAD biosynthesis. In a mouse LLC tumor model overexpressing NAMPT and PD-L1, significantly suppressed tumor progression through dual mechanisms involving tumor metabolism disruption and tumor immune microenvironment activation. These findings highlight as a promising lead compound for the development of next-generation antitumor agents that simultaneously target tumor immunoevasion and metabolism.
Monoamine oxidase B (MAO-B), upregulated in reactive astrogliosis, represents a promising positron emission tomography (PET) target for neurodegenerative disorders. In this study, 37 fluorinated indazole carboxamide deri...Monoamine oxidase B (MAO-B), upregulated in reactive astrogliosis, represents a promising positron emission tomography (PET) target for neurodegenerative disorders. In this study, 37 fluorinated indazole carboxamide derivatives were designed and synthesized as MAO-B inhibitors. Among them, compound showed outstanding MAO-B inhibitory activity (IC = 0.07 nM) and excellent selectivity over MAO-A. Automated radiosynthesis offered [F] with high molar activity (135.8 GBq/μmol) and radiochemical yield (36.5%, decay-corrected to end-of-bombardment). Dynamic PET imaging revealed efficient blood-brain barrier penetration of [F] (SUV = 1.40 in rat), with specific binding confirmed by selegiline blocking. autoradiography revealed region-specific binding of [F] to MAO-B in rat brain. Metabolism studies showed that 62 ± 4% of brain radioactivity remained as the parent fraction at 30 min post-injection. Altogether, this work provides a novel MAO-B PET tracer based on an indazole carboxamide scaffold, potentially providing inspiration for future tracer development.
Interleukin-1 receptor-associated kinase 4 (IRAK4) represents an attractive therapeutic target in inflammatory and oncological indications due to its pivotal role as a signaling mediator downstream of TLR and IL-1R. We p...Interleukin-1 receptor-associated kinase 4 (IRAK4) represents an attractive therapeutic target in inflammatory and oncological indications due to its pivotal role as a signaling mediator downstream of TLR and IL-1R. We previously reported a series of spirocyclic IRAK4 degraders and identified APH02174 as a selective, orally bioavailable degrader. However, its hERG inhibition, hepatocyte stability, and variable cross-species PK inspired further structural optimization. The optimization gave eight PROTACs (-, and ) which exhibited high IRAK4 degradation ability (DC ≤ 2 nM, DC ≤ 10 nM, and ≥ 85%) . exhibited favorable PK parameters, low hERG inhibition, improved hepatocyte stability, and pronounced anti-inflammatory efficacy in animal models. Dose range-finding studies in rat and dog models supported its safety profile. Based on these improvements in safety, cross-species PK, efficacy, and preliminary toxicology, was selected as a preclinical candidate for GLP toxicological studies.
Pre-mRNA splicing is a process of removing introns from precursor RNA and joining exons to form mature RNA for protein translation. The processes are classified into constitutive and alternative splicing, with which mult...Pre-mRNA splicing is a process of removing introns from precursor RNA and joining exons to form mature RNA for protein translation. The processes are classified into constitutive and alternative splicing, with which multiple mRNA transcripts are generated from a single coding gene to expand protein diversity. Frequent mutations or abnormal expressions of some splicing factors dysregulate splicing in a subset of transcripts, leading to aberrant gene expression patterns. Notable splicing factors, including SF3B1, U2AF1, SRSF2, and RBM39, are considered potential drug targets for various cancers. Advancement in the discovery and development of small molecules targeting specific splicing factors has become a new treatment modality for cancer therapy. Here, we review recurrent mutations and dysregulated expressions of splicing factors in cancers and provide our perspective on recent developments of small-molecule compounds targeting splicing factors and the functional assays that facilitate hit/lead discovery for development to treat cancer.
Nicotinic acetylcholine receptors (nAChRs) containing the α6 subunit represent a promising therapeutic target for nicotine addiction. By employing structure-guided optimization of these α-conotoxin leads, we developed po...Nicotinic acetylcholine receptors (nAChRs) containing the α6 subunit represent a promising therapeutic target for nicotine addiction. By employing structure-guided optimization of these α-conotoxin leads, we developed potent and selective inhibitors of the human α6β4 nAChR subtype, achieving single-digit nanomolar potency and >40-fold selectivity over related nAChR subtypes. The lead compound demonstrated enhanced enzyme stability and, in a nicotine-induced conditioned place preference model in mice, effectively attenuated addictive behaviors. These results present a novel, potent and selective α6β4 nAChR antagonist with significant potential as a therapeutic candidate for nicotine use disorder.
A key challenge in organophosphorus (OP) research is to develop efficient means to decrease OP levels after exposure. Scavenging, capturing, or degrading circulating OPs could minimize OP penetration into the brain and...A key challenge in organophosphorus (OP) research is to develop efficient means to decrease OP levels after exposure. Scavenging, capturing, or degrading circulating OPs could minimize OP penetration into the brain and other organs, and decrease toxicity. OPs produce acute effects primarily through acetylcholinesterase (AChE) inhibition, leading to neurotoxic events. Current standard of care (SOC) for OP exposure includes the AChE reactivator 2-PAM as well as atropine, and midazolam or diazepam. However, SOC therapeutics only address AChE symptomatic toxicity but do not increase clearance. SOC does not degrade OPs. Using haptens designed employing transition state analogy, we procured and characterized monoclonal antibodies (mAbs) that, in the presence of 2-PAM, increased the hydrolysis of paraoxon (POX) and promoted degradation of nerve agent mimics. The ability of these novel antibodies to combine with 2-PAM and selectively recognize and degrade OPs suggests that they could provide a new approach to decrease OP-induced toxicity.
Developing photocages within the phototherapeutic window is crucial for efficient photopharmacology regulation, yet it poses significant challenges. Herein, we present a photocage prodrug of doxorubicin-methylene blue (D...Developing photocages within the phototherapeutic window is crucial for efficient photopharmacology regulation, yet it poses significant challenges. Herein, we present a photocage prodrug of doxorubicin-methylene blue (DOX-MB) linked by a urea bond, offering spatiotemporal therapeutic selectivity and reduced systemic toxicity. In particular, this photocage prodrug remains stable under physiological conditions, exhibiting reduced cytotoxicity and quenched fluorescence. Upon 660 nm laser irradiation, DOX-MB generates reactive oxygen species that selectively cleave the urea linkage, facilitating the concurrent release of an active pharmaceutical ingredient. This photoactivatable cleavage of DOX-MB allows for precise chemotherapy and photodynamic therapy and restores MB FL for real-time monitoring as well. To achieve localized tumor therapy, the DOX-MB prodrug was encapsulated into a thermosensitive hydrogel, further minimizing the systemic exposure. In vivo studies showed that this integrated platform achieved a tumor inhibition rate of 80.3%. Collectively, this study presents a precision-engineered photoactivatable prodrug that enables synergistic photochemotherapy.