The efficacy of FLT3 inhibitors in acute myeloid leukemia (AML) is severely limited by resistance mutations, particularly the recalcitrant gatekeeper F691L and activation loop D835V/Y variants. Herein, through the structure-guided optimization of our previously reported lead SILA-123, we identified FYJ-195, a highly potent type II FLT3 inhibitor capable of overcoming these recalcitrant mutants. FYJ-195 exhibited single-digit nanomolar potency against Ba/F3-FLT3-ITD-F691L (IC50 = 9.09 nM) and subnanomolar activity against Ba/F3-FLT3-ITD-D835Y (IC50 = 4.64 nM) and Ba/F3-FLT3-ITD-D835V (IC50 = 0.76 nM). In vivo, FYJ-195 induced profound tumor regression (TGI = 125%) in the MV4-11 xenograft model (10 mg/kg) and achieved robust tumor growth suppression (TGI = 68.6%) in the Ba/F3-FLT3-ITD-F691L model (50 mg/kg), where quizartinib was ineffective. Mechanistic studies confirmed that FYJ-195 effectively blocked FLT3 signaling and induced apoptosis without observable toxicity. Collectively, FYJ-195 represents a promising lead candidate for drug-resistant AML.

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related mortality worldwide, hindered by drug resistance and metastasis. Epithelial-mesenchymal transition (EMT) drives NSCLC progression and therapy resistance, but few effective EMT-targeting strategies exist. Acteoside, a phenylethanoid glycoside has shown promising antitumor activity, but its specific mechanisms in NSCLC remain unclear. This study aimed to define the anti-NSCLC potency of acteoside, uncover its mechanism, and appraise its translational value. In vitro proliferation assays and distinct xenografts (monotherapy and Baihe Gujin Decoction) quantified efficacy. Integrative transcriptomics, proteomics, western blot, molecular dynamics, surface plasmon resonance (SPR) assay and knockdown experiment mapped the mode of action, while in vivo HPLC-MS/MS-based pharmacokinetic (PK) profiling gauged bioavailability. Functional validation supported that acteoside inhibited NSCLC cell migration in Transwell assays. Acteoside, identified as the key bioactive compound in Baihe Gujin Decoction, demonstrated potent anti-NSCLC efficacy both in vitro and in vivo. It inhibited A549 cell proliferation (IC50 = 3.79 µM), induced apoptosis and G0/G1 arrest, and significantly suppressed tumor growth in xenograft models without toxicity. Pharmacokinetic studies revealed acteoside acts as a prodrug, rapidly converting to caffeic acid. Multi-omics, IL-17 pathway perturbation assay and molecular analyses supported that acteoside upregulates E-cadherin (CDH1) via IL-17 pathway inhibition, with stable binding to the E-cadherin dimer interface validated by molecular dynamics, SPR, knockdown experiment and Western blot. Transwell assays functionally validated EMT modulation and migration inhibition. These findings provide evidence for acteoside as a novel small-molecule CDH1 modulator for NSCLC treatment. Acteoside is an antitumor agent that suppresses cancer cell proliferation and induces tumor regression in preclinical models, with no overt toxicity observed at the tested doses. Mechanistically, acteoside upregulates E-cadherin (CDH1) expression and directly binds to the CDH1 protein with measurable affinity, while also modulating the IL-17 signaling pathway. Pharmacokinetic studies indicate that acteoside undergoes extensive metabolism after oral administration, with its metabolite caffeic acid reaching higher systemic exposure, suggesting a potential prodrug characteristic. These findings position acteoside as a promising candidate for CDH1-targeted therapeutic strategies in non-small cell lung cancer (NSCLC), warranting further investigation.

Herein, we present a database, KLSD, which is a curated resource of 787 213 small-molecule kinase inhibitors annotated with 1.8 M quantitative activity records across 428 human kinases, emphasizing selectivity and polypharmacology. Moreover, we introduce a dual-task ensemble that simultaneously regresses pAct and computes selectivity scores. The core is a multibranch residual multilayer perceptron (MLP) whose branches are kinase-specific; this is augmented by SVM, RF, XGBoost, CNN, GCN, GAT, RGCN and VAE-enhanced graph nets. Continuous potency labels replace categorical classes to improve resolution. Benchmarked on the JAK family (JAK1/2/3, TYK2), the ensemble achieves classification accuracies of ≥0.84 for each kinase and 0.98 overall, demonstrating strong generalizability. KLSD and models are freely available at http://ai.njucm.edu.cn:8080.

Two series of bufalin derivatives including three compounds were designed, synthesized, and evaluated for their antiproliferative activity against A549 (non-small cell lung cancer) and HeLa (cervical cancer) cells. Structure-activity relationship (SAR) analysis revealed that methylation of the C3β-hydroxy group or replacement of the C17 six-membered unsaturated lactone ring with a five-membered lactone ring reduced the antitumor activity. These findings indicate that both the C3β-OH group and the intact C17 lactone moiety are essential pharmacophoric elements for the antitumor activity of bufalin.

Background The cyclic GMP-AMP synthase/stimulator of interferon genes (cGAS-STING) pathway mediates anti-tumor immunity by sensing cytosolic DNA and producing type I interferons, but is frequently suppressed in multiple myeloma (MM), enabling immune evasion. Dexamethasone (DXM), a cornerstone glucocorticoid in MM therapy, is paradoxically combined with immunotherapies despite its immunosuppressive reputation. The mechanisms by which DXM and its glucocorticoid receptor (GR) modulate the MM immune microenvironment remain elusive. Methods We analyzed the tumor immune microenvironment using single-cell RNA sequencing in 5TMM3VT murine models. STING-knockout mice were employed for genetic validation, and the PP1 inhibitor salubrinal for pharmacological studies. Protein-protein interactions were examined by co-immunoprecipitation, mass spectrometry, and immunofluorescence. NK cell cytotoxicity and macrophage polarization were assessed using co-culture systems with flow cytometry, quantitative real-time PCR, and ELISA. Results We identified that GR constitutively recruits the phosphatase PPP1CB to dephosphorylate and inactivate the TBK1-IRF3 complex, thereby suppressing cGAS-STING signaling in MM cells under basal conditions. As the specific ligand of GR, DXM rapidly binds to GR, leading to the disruption of the GR-PPP1CB-TBK1-IRF3 complex, which in turn reactivates STING signaling and induces interferon-stimulated gene expression. Single-cell analysis revealed that DXM remodeled the immune microenvironment, enhancing NK cell cytotoxicity, and promoting M1-like macrophage polarization. These immunostimulatory effects were abrogated in STING-deficient models. Pharmacological inhibition of PP1 with salubrinal recapitulated DXM’s effects, synergized with bortezomib to prolong survival in vivo , and enhanced daratumumab-mediated antibody-dependent cellular cytotoxicity (ADCC). Conclusions This study uncovers a glucocorticoid-sensitive inhibitory axis wherein GR-PPP1CB constitutively suppresses STING-TBK1-IRF3 signaling in MM. DXM relieves this suppression, reactivating innate anti-tumor immunity. PP1 inhibition represents a promising therapeutic strategy to enhance anti-myeloma immunity and overcome immune evasion.

FLT3 is a validated therapeutic target in acute myeloid leukemia (AML), yet resistance mutations frequently limit current inhibitors. Here, we report a series of 6-methylisoxazolo[5,4-b]pyridin-3-amines that covalently target Cys807, a previously unexploited nucleophilic residue within the FLT3 kinase domain. Compound 18 (FLC-8) potently inhibited FLT3-WT (IC50 = 10.2 nM) and clinically relevant mutants G697R (IC50 = 11.6 nM) and N676D (IC50 = 24.1 nM). Covalent engagement of Cys807 was confirmed by mass spectrometry, peptide mapping, and loss of activity upon C807S mutation. FLC-8 suppressed FLT3-mediated STAT5, AKT, and ERK signaling and induced apoptosis in AML cells while maintaining low-nanomolar potency over 72 h. Kinome profiling revealed a narrow inhibition spectrum. In vivo, FLC-8 inhibited MV4-11 xenograft growth (TGI: 136-178% at 10-50 mg/kg) without overt toxicity. These findings identify Cys807 as a covalent binding hotspot in FLT3 and establish FLC-8 as a promising scaffold for next-generation FLT3 inhibitor development.

Chromatin abnormalities are a hallmark of cancer, but the role of the chromatin remodeler CHD4 in multiple myeloma (MM) remains unclear. This study aims to elucidate how CHD4 drives MM progression and to explore targeted therapeutic strategies. Using CHD4 overexpression and knockdown models in MM cell lines, we found that elevated CHD4 correlates with poor prognosis and increased proliferation. Mechanistically, we performed chromatin profiling and luciferase reporter assays to show that the ATPase domain of CHD4 resolves G-quadruplex structures in the c-Myc promoter, thereby enhancing chromatin accessibility. Co-immunoprecipitation (Co-IP) and chromatin immunoprecipitation (ChIP) further revealed that CHD4 stabilizes c-Myc activity through liquid-liquid phase separation (LLPS), increasing c-Myc chromatin retention and promoter occupancy. To identify CHD4 targets, we integrated RNA-seq with chromatin profiling. Structure-based drug design identified luteolin 7-O-glucuronide (LUT) as an inhibitor of CHD4's ATPase domain and idarubicin (IDA) as an inhibitor of its chromatin-interaction domain; combination treatment synergistically suppressed MM proliferation in adoptive B-cell transfer, xenograft, and 5TMM3VT mouse models. Our findings establish CHD4 as a key oncogenic driver in MM and propose a LUT/IDA combination as a precision medicine strategy, advancing the understanding of chromatin remodeling in cancer.

RET fusions and activating mutations drive multiple human cancers, while resistance mutations limit the efficacy of current selective RET inhibitors. Here, we report CN-3, a potent RET inhibitor active against clinically relevant mutants, including solvent-front (G810R/S/C), gatekeeper (V804M), hinge (Y806H), and catalytic loop (M918T) variants. CN-3 inhibited all tested RET mutants (IC50 < 5 nM) and selectively suppressed proliferation of RET-driven cellular models, including TT (IC50 = 2.48 ± 0.78 nM) and LC-2/ad (IC50 = 17.05 ± 4.90 nM) cells, as well as Ba/F3 cells expressing RET fusions or mutations, without affecting RET-independent or normal cells. Mechanistically, CN-3 blocked RET autophosphorylation and downstream SHC/AKT/ERK signaling, inducing G0-G1 arrest and apoptosis. In RET-driven xenografts, CN-3 showed dose-dependent antitumor efficacy with good tolerability. Kinase profiling revealed moderate selectivity, with off-target activity mainly restricted to a limited group of receptor tyrosine kinases. These results support CN-3 as a promising lead for next-generation RET-targeted therapies.

Retinoic acid receptor-related orphan receptor γ (RORγ) is a key transcriptional regulator of the T helper 17 cell and has emerged as a promising therapeutic target for autoimmune diseases. To identify novel orthosteric inhibitors of RORγ, we designed and synthesized ten fluorescent probes targeting the RORγ orthosteric site. Among them, probe 19g exhibited the best performance (Kd = 252 nM, fluorescence quantum yield = 29.7%) and was subsequently employed to establish a fluorescence polarization (FP)-based assay for screening natural orthosteric binders. Using this optimized FP system, the natural polyphenolic compounds tannic acid (FP Ki = 250 ± 13 nM) and epigallocatechin gallate (EGCG, FP Ki = 506 ± 8 nM) were identified as RORγ ligands. Both compounds exhibited significant inhibitory activity against RORγ, as assessed by RORγ-Gal4 reporter, qPCR and CESTA assays. Furthermore, in the psoriatic-like mouse model, EGCG can effectively alleviate psoriatic-like skin lesions. Collectively, the fluorescent probes and FP-based screening platform provide convenient tools for the discovery and mechanistic investigation of RORγ orthosteric inhibitors. Moreover, the identification of RORγ as a molecular target of tannic acid and EGCG offers new insights into the anti-inflammatory mechanisms of these natural compounds.

Luteolin, a flavonoid present in botanical drugs, plants, and dietary sources, has demonstrated anticancer properties against various tumors, yet its role in diffuse large B-cell lymphoma (DLBCL) remains unclear. This study aimed to uncover the molecular mechanism of luteolin in DLBCL treatment using a combination of in vitro and in vivo experiments and computational analysis. Human DLBCL cell lines U2932 and OCI-LY10 were utilized to assess luteolin’s impact on cell growth, apoptosis, cell cycle progression, and the modulation of JAK2/STAT3 pathway proteins. In vivo, a U2932 tumor-bearing nude mice model was employed to evaluate luteolin’s antitumor efficacy and its effects on JAK2/STAT3 pathway protein expression. Additionally, molecular dynamics simulations were conducted to explore the interaction between luteolin and JAK2. The findings revealed that luteolin significantly suppressed cell proliferation, induced apoptosis, and arrested the cell cycle at the G2/M phase in both cell lines. In the mouse model, luteolin effectively inhibited tumor growth and downregulated the expression of phosphorylated JAK2 and STAT3 without altering the total protein levels of JAK2 and STAT3. Computational analysis indicated stable binding of luteolin to JAK2. Collectively, these results suggest that luteolin’s anti-DLBCL activity may be mediated through the regulation of the JAK2/STAT3 signaling pathway, positioning it as a potential therapeutic agent for DLBCL.

Most cancers are currently incurable, partly due to abnormal post-translational modifications (PTMs). In this study, we initially used multiple myeloma (MM) as a working model and found that SUMOylation activating enzyme subunit 1 (SAE1) promotes the malignancy of MM. Through proteome microarray analysis, SAE1 was identified as a potential target for bioactive colcemid or its derivative colchicine. Elevated levels of SAE1 were associated with poor clinical survival and increased MM proliferation in vitro and in vivo. Additionally, SAE1 directly SUMOylated and upregulated the total protein expression of p27, leading to LLPS-mediated nuclear export of p27. Our study also demonstrated the involvement of SAE1 in other types of cancer cells, and provided the first monomer crystal structure of SAE1 and its key binding model with colchicine. Colchicine also showed promising results in the Patient-Derived Tumor Xenograft (PDX) model. Furthermore, a controlled clinical trial with 56 MM patients demonstrated the clinical efficacy of colchicine. Our findings reveal a novel mechanism by which tumor cells evade p27-induced cellular growth arrest through p27 SUMOylation-mediated nuclear export. SAE1 may serve as a promising therapeutic target, and colchicine may be a potential treatment option for multiple types of cancer in clinical settings.

Aims Transcriptional dysregulation by aberrant transcription factors (TFs) is a key driver of drug resistance. Resistance to adriamycin (ADR) frequently develops following first-line treatment for multiple myeloma (MM). This study aims to identify novel TFs associated with ADR resistance in MM and to elucidate their underlying mechanisms. Methods We employed a protein chip assay with FITC-labeled celastrol and identified the deformed epidermal autoregulatory factor 1 (DEAF1) as a potential target in MM. High-throughput sequencing was performed to identify DEAF1 downstream targets. Both in vivo and in vitro models were utilized to delineate the role of DEAF1 in MM cell proliferation and ADR resistance. Results High DEAF1 expression was associated with poor prognosis in MM patients, and was found to promote MM cell proliferation and induce ADR resistance. Mechanistically, DEAF1 directly binds to the RAD50 promoter via its SAND domain, upregulating RAD50 expression and consequently activating the ATM pathway. Furthermore, DEAF1 recruited AP-2-alpha (AP-2α) through its MYND domain, leading to the downregulation of tyrosine-protein kinase Fer (FER). This downregulation impaired FER-mediated phosphorylation of GSDME, which is known to enhance the cleavage efficiency of GSDME by caspase-3. Additionally, celastrol synergized with ADR to inhibit MM cell viability by disrupting the binding of DEAF1 to the promoters of its target genes. Conclusions Our findings demonstrate that DEAF1 attenuates ADR-induced apoptosis and pyroptosis in MM by enhancing DNA damage repair and suppressing GSDME cleavage via the FER/GSDME axis. This study provides a novel therapeutic target for the treatment of MM.

This study presents an innovative computational method, combined with experimental validation, to predict and elucidate the metabolic pathways of SILA-123, a novel FLT3 inhibitor. Using UFLC/Q-TOF MS, we identified 21 metabolites generated through key reactions (oxidation, reduction, hydrolysis, cleavage, deamination, and glucuronidation). Advanced computational techniques were applied to identify key metabolic enzymes, with results confirmed experimentally. Our findings represent a significant advancement in the field of metabolic prediction. By integrating computational methods with experimental data, we have established a robust framework that can be applied to other therapeutic compounds. This approach not only enhances our understanding of SILA-123's metabolic pathways but also provides a novel strategy for guiding metabolic prediction.

Danatinib, a brand-new compound synthesized in our laboratory for the treatment of acute myeloid leukemia (AML), demonstrates remarkable antitumor activity. However, the pharmacokinetic profile of Danatinib in mice was short with its half-life was only 0.476 h. To address this issue and optimize its therapeutic efficacy, this study investigated the metabolic profiles of Danatinib in mice, both in vitro and in vivo, using ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) technology. In the in vitro study, Danatinib was analyzed using mouse liver microsomes, resulting in the identification of eight metabolites. In the in vivo study, Danatinib was administered orally to mice at 20 mg/kg; the samples of plasma, bile, feces, and urine were collected and analyzed, leading to the identification of 34 metabolites. The results showed that those metabolites were formed through various metabolic reactions including hydroxylation, carboxylation, acetylation, hydrogenation, and glucuronidation. This study provides a systematic investigation of the metabolism of Danatinib, offering valuable information for further structural modification to improve its pharmacokinetic profiles.

One of the major limitations of cancer therapy is the emergence of drug resistance. This review amis to provide a focused analysis of the multifactorial mechanisms underlying therapy resistance,with an emphasis on actionable insights for developing novel therapeutic strategies. It concisely outlines key factors contributing to therapy resistance, including drug delivery barriers, cancer stem cells (CSCs), epithelial-mesenchymal transition (EMT), cancer heterogeneity, tumor microenvironment (TME), genetic mutations, and alterlations in gene expression. Additionally, we explore how tumors evade targeted therapies through pathway-specific mechanisms that restore disrupted signaling pathways. The review critically evaluates innovative strategies designed to sensitize resistant tumor cells, such as targeted protein dedgradation, antibody-drug conjugates, structure-based drug design, allosteric drugs, multitarget drugs, nanomedicine and others We also highlight the importance of understanding the pharmacological actions of these agents and their integration into treatment regimens. By synthesizing current knowledge and identifying gaps in our understanding, this review aims to guide future research and improve patient outcomes in cancer therapy.

A concise and scalable semi-synthesis of (+)-digitoxigenin ( 1 ) has been accomplished in nine linear steps. This route features the construction of the C(15)=C(16) double bond in intermediate 6 via selective C16 bromination followed by elimination. Subsequesntly, a single-step SeO2 oxidation directly installed the C14-beta-hydroxyl group, producing 7 . This strategy avoids precious metals, greatly reduces synthesis costs, and can be extended to the synthesis of related cardiotonic steroids.

Background: Alzheimer's disease (AD), a chronic neurodegenerative disorder predominantly occurs among the elderly, is the leading cause of dementia. The accumulation of beta-amyloid (A beta) is considered the main pathogenies of AD, and beta-site APP-cleaving enzyme 1 (BACE1) plays an important role in the formulation of A beta. Objective: In order to find a new scaffold as BACE1 inhibitors, a series of novel 2-amino-1-phenyl-benzimidazole derivatives were designed and synthesized in this work. Methods: Using our previous L-5 as a lead compound, we applied a scaffold hopping method and merged 2-amino-1-methyl-4-phenyl-1H-imidazol-5 (4H)-one into benzimidazole, so a novel class of BACE1 inhibitors T1 similar to T20 with the structure of 2-amino-1-phenyl-benzimidazole were designed and synthesized. Results: The biological activity evaluation indicated that the target compounds showed inhibitory activities against BACE1, with T14 being the most potent (IC50 = 0.45 mu M), it also exhibited good logP value and tPSA. The docking studies indicated that compound T14 could form important hydrogen bonds with Asp289 and Asp93. Conclusion: Compound T14 could be used as a potential BACE1 inhibitor for further modification to treat AD.