Nine isopentenyl flavonoids (1–7), including ten new compounds (1–4 and 5a/5b–6a/6b), were obtained from Daphne giraldii based on ion identity molecular networking (IIMN).

The first total synthesis of scarce alkaloid indigotinoline A from Isatis indigotica was achieved in six steps from a known carboxylic acid (13% overall yield). The distinctive 6/5/6/5/6 N,O-heterocycle was efficiently assembled via a one-pot triple cascade, Povarov-type cyclization, intramolecular substitution, and visible-light-mediated oxidative rearrangement, followed by bioinspired N-oxidation/1,2-migration. This synthesis also prompted re-examination of proposed biosynthetic precursor isatindigotindoline B, leading to revision of its stereochemistry based on spectroscopic analysis.

Brusatol (BST), a quassinoid triterpenoid exhibits remarkable anti-liver cancer activity. However, the high toxicity and the lack of specific tumor targeting capabilities limit its antiliver cancer efficacy. To address these limitations, we developed a GLUT1-targeted and esterase-responsive prodrug liposome strategy that integrates metabolic targeting with intracellular environment-triggered drug release. Esterase-sensitive prodrugs were synthesized by conjugating BST with long-chain fatty acids, then encapsulated into liposomes modified with a novel GLUT1-targeting head group to yield the prodrug liposome formulation (9C-2G-lipo). Our results showed that 9C-2G-lipo significantly enhanced the tumor growth inhibition rate from 36.96 % (free BST) to 68.37 % at an equivalent dose, while reducing systemic toxicity, as reflected by a two-fold increase in median lethal dose (LD50 = 10.21 mg/kg) compared to free BST (LD50 = 5.02 mg/kg). Meanwhile, the resulting 9C-2G-lipo showed 72.31 % enhanced tumor accumulation in Hep3B xenografts via GLUT1-mediated tumor targeting, cumulative released 74.15 % active BST under tumor-mimetic conditions (pH 5.2, esterase-rich). Mechanistically, 9C-2G-lipo induced G2/M cell cycle arrest and promoted apoptosis by regulating the HSP90/EGFR/MAPK pathway. The study highlighted integrating tumor-metabolic targeting and microenvironment responsiveness strategy as a paradigm for transforming toxic natural products into safe, efficacious nanomedicines with an improved therapeutic index for liver cancer therapy.

BACKGROUND:Brucea javanica (L.) Merr. (B. javanica) is a medicinal plant with a long history of clinical application and proven antitumor activity. Its oil emulsion (BJOE) is widely used as a cancer adjuvant therapy. However, a comprehensive understanding of its chemical composition, multi-component synergistic mechanism, key antitumor signaling pathways, formulation optimization challenges, clinical translation bottlenecks, and safety issues remain insufficient, which greatly limits its rational development and precise clinical application. PURPOSE:This review aims to systematically summarize the chemical composition, pharmacological mechanism, antitumor properties, formulation progress, clinical efficacy and safety of B. javanica, and critically point out the current research gaps and key challenges in basic and applied research. STUDY DESIGN AND METHODS:Keywords related to B. javanica, quassinoids, structure-activity relationships, pharmacology, formulations, and clinical aspects to include published studies from 2008-2025 were searched in the following databases: SciFinder, Web of Science, PubMed, China National Knowledge Infrastructure (CNKI) and Wanfang database. Relevant literature in both Chinese and English obtained from the above search was incorporated into this review. RESULTS:Over 100 quassinoids, 48 triterpenoids, and 27 alkaloids have been identified from B. javanica. As the core antitumor components, quassinoids exert pharmacological activities through the regulation of PI3K/AKT, MAPK, NF-κB and various other vital signaling cascades. Targeted nano-formulations can enhance tumor accumulation, but large-scale production is difficult and stability is insufficient. Clinically, BJOE can improve the efficacy of chemo-radiotherapy, but carries risks of allergic reactions and multi-system injury. CONCLUSION:This review clarifies the key antitumor components, signaling pathways, delivery strategies and safety profiles of B. javanica. Despite its significant antitumor potential, instability in quality, unclear synergistic mechanisms, unstable delivery systems, and safety issues remain major challenges. This study provides comprehensive evidence base and identifies key research gaps to support the further rational development of therapies based on B. javanica.

The first hemiterpene-quassinoid adducts, bruquass A and B (1 and 2), were rapidly isolated and identified from Brucea javanica using an integrated analytical strategy. They possessed unusual carbon skeletons formed by the coupling of quassinoids with hemiterpene units via vinylogous aldol reactions. Their structural configurations were determined through comprehensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. Plausible biosynthetic pathways for 1 and 2 were proposed, and guided by these biogenetic insights, the biomimetic synthesis of compound 1 was successfully achieved. Furthermore, compounds 1 and 2 exhibited significant antifeedant activity against Plutella xylostella. The bioactivity assessment results open up the prospects of 1 and 2 as a promising new class of botanical insecticide.

In China, Elephantopus scaber L. and Elephantopus tomentosus L., two traditional Chinese medicinal herbs commonly known as "Ku-di-dan", are widely used. Moreover, they have demonstrated anti-hepatoma effects and hepatoprotective properties in experimental studies. Nonetheless, according to existing literature, E. scaber and E. tomentosus encompass compounds with similar but notably different chemical structures. Therefore, investigating the differences in secondary metabolites between E. scaber and E. tomentosus and elucidating their distinct anti-hepatocellular carcinoma activities is of great importance. This study employed an integrated approach combining untargeted metabolomics with bioactivity assays to effectively differentiate between the congeneric plant species E. scaber and E. tomentosus. To improve the accuracy of characterizing and identifying key metabolites, an in-house database and data screening methods were employed. The research findings revealed that thirty major secondary metabolites were detected in E. tomentosus and E. scaber extracts through untargeted metabolomics and data screening. Subsequently, eight significant differential metabolites were identified through multivariate statistical analyses. Thereafter, twelve compounds were isolated from these two plants and evaluated for their anti-hepatocellular activity, revealing that compounds 5-8 exhibited significant inhibitory effects on HepG2 and Hep3B cells. Consistent results were also observed in the tests conducted on the crude extracts of both E. scaber and E. tomentosus. This study demonstrates that E. scaber and E. tomentosus exhibit distinct secondary metabolite profiles and differential anti-hepatocellular carcinoma activities.

Ischemic stroke (IS) is a major global cause of morbidity and mortality, highlighting the need for safe and effective treatments. Diospyros kaki leaves are a key ingredient in the clinical drug NaoXinQing and the health beverage persimmon leaf tea, yet their active components remain poorly defined. In this study, 16 compounds, including two new terpenoids (1, 3) and 14 known monoterpenoids (2, 4-16), were isolated from D. kaki leaves using an MS/MS molecular networking approach. Their structures were elucidated via NMR, quantum chemical NMR, and ECD calculations. Neuroprotective screening in OGD/R-injured HT22 cells showed that compounds 7 and 14 exhibited significant activity. They reduced apoptosis and intracellular ROS accumulation. Mechanistic studies, including network pharmacology, molecular docking, molecular dynamics, and Western blotting, confirmed that 7 and 14 protect neurons by inhibiting the JAK2/STAT3 signaling pathway. This work reveals terpenoids from D. kaki leaves as potential anti-ischemic agents, clarifies the active material basis of NaoXinQing and persimmon leaf tea, and supports further development of this medicinal resource.

Objective: Daphnegiravone D (DGD), a prenylated flavonoid from Daphne giraldii Nitsche, a plant whose roots and stem are used in Chinese medicine, exerts an inhibitory effect on liver cancer cells. The protein kinase ataxia telangiectasia-mutated and Rad3-related (ATR) is an important component of the DNA damage response, and its inhibition enhances the sensitivity of some cancer cells to DNA-damaging drugs. Oxaliplatin (OXA) is a third-generation, platinum-based anticancer drug that exerts its inhibitory effects on liver cancer cells by inhibiting DNA synthesis. Owing to the fact that most patients develop drug resistance, finding novel treatments for liver cancer is urgent. In a previous study, DGD was shown to exert anti-tumor effects as a potential ATR kinase inhibitor. This study investigated the mechanism of action of DGD and its combination with OXA in liver cancer. Methods: HepG2 and Hep3B cells were used to evaluate the inhibitory effects of DGD on liver cancer. Cellular thermal shift assay, co-immunoprecipitation, and western blotting (WB) were used to confirm the impact of DGD on DNA damage. The methyl thiazolyl tetrazolium colorimetric method and the results were used to calculate the drug synergy score. Acridine orange/ethidium bromide staining, Annexin V/Propidium iodide staining, reactive oxygen species-related staining, and WB were performed to assess the efficacy of the combination of DGD and OXA. In vivo tumor xenograft model in nude mice was used to investigate the efficacy of this combination. Hematoxylin-eosin staining and immunohistochemistry were performed to observe the condition of tissues in vivo . Results: DGD inhibited liver cancer cells by affecting the formation of the ATR-ATRIP complex and downregulating the activator protein TopBP1. We found that the combination of DGD and OXA synergistically inhibited the growth of liver cancer cells and reduced the toxicity of OXA to normal hepatocytes. This synergistic effect was mediated by the induction of apoptosis, mitochondrial dysfunction, and oxidative stress. Further experiments suggested that DGD may suppress the DNA damage response by inhibiting the ATR pathway, which, in turn, enhances the effect of OXA on DNA damage. Consistent with the above results, DGD enhanced the anticancer effect and moderated the side effects of OXA in vivo . Conclusion: Our results showed that DGD greatly enhanced the anti-tumor effect of the chemotherapeutic drug OXA via the ATR pathway in liver cancer, both in vitro and in vivo . Graphical Abstract: http://links.lww.com/AHM/A231

Signal transducer and activator of transcription 3 (STAT3) is a critical oncogenic driver hyperactivated in approximately 70% of human cancers, especially colorectal cancer (CRC), making it an attractive therapeutic target. Currently, the discovery of STAT3 inhibitors is primarily focused on the SH2 domain, however, there are still problems such as insufficient inhibitory activity and poor clinical efficacy. Targeting the DNA-binding domain (DBD) is a direct and effective strategy for inhibiting STAT3 function; but due to the lack of suitable targeting sites and design methods, reported compounds and their mechanisms of action are limited. Here, we identified a germacrane-type sesquiterpene lactone, SCP-7, from a natural product library through multi-dimensions progressive screening. SCP-7 exhibited potent binding affinity to the STAT3-DBD, with a Kd of 795 nM. Mechanistically, SCP-7 covalently modifies Cys328 and Cys367 of STAT3, as confirmed by mass spectrometry and mutational analysis, with Cys328 serving as the primary binding site and effectively inhibit the DNA-binding activity of STAT3. In CRC cells (HCT116 and HT29), SCP-7 effectively reduced p-STAT3 and c-Myc levels, and inhibited proliferation, migration, and colony formation. Moreover, SCP-7 induced G2/M cell cycle arrest, triggered mitochondrial dysfunction, elevated reactive oxygen species levels, and promoted apoptosis via modulation of BAX, Bcl-2, Mcl-1 and survivin. In a mouse xenograft model, SCP-7 significantly suppressed tumor growth, exhibiting activity comparable to that of oxaliplatin but with a markedly improved safety profile. Collectively, our findings identify SCP-7 as a novel covalent inhibitor targeting the STAT3-DBD with potent anti-CRC activity in vitro and in vivo, providing a promising lead compound for STAT3-targeted cancer therapy.

The natural products ciquitin A and ciquitin B exhibit two distinct sets of NMR signals in solution, a phenomenon previously attributed to an equilibrium between its ring-open and ring-closed hemiacetal isomers. Through a comprehensive re-evaluation of NMR data, complemented by variable-temperature NMR experiments, potential energy surface (PES) scanning, and magnetic shielding constant calculations, we have redefined ciquitin A as a natural pseudo-resonance structure. Prior to successful crystallization for X-ray analysis, structural identification remained challenging when facing two sets of NMR spectra. However, conversion of any pseudo-resonance structure (like ciquitin A, a hemiacetal type) to its corresponding acetal derivatives (in this case (9S)-naproxyl-ciquitin A) resulted in a single set, thereby enabling clear identification of the structure. This work represents one of the most unequivocal examples of such a structure discovered in nature.

As a traditional medicinal-food ingredient, pepper roots were investigated for bioactive dimeric amide alkaloids. Eighteen pairs of novel anti-inflammatory dimeric amide alkaloids (1a/1b-18a/18b) and two known analogues (19 and 20) were isolated under the guidance of a feature-guided molecular networking (FGMN) strategy, with their structures established by extensive spectroscopic analyses and X-ray diffraction. A visible light photocatalytic method enabled the sustainable synthesis of the dimeric scaffold directly from piperine, a key component of Piper nigrum. In vitro assays demonstrated significant inhibition of NO production in a LPS-induced RAW 264.7 macrophage model. Additionally, dimeric amide alkaloid 21 significantly alleviated skin inflammation, demonstrating bioactivity from a food-derived compound. Overall, this study highlights the potential of P. nigrum roots as a sustainable source of functional alkaloids and establishes a photocatalytic route to skin-health-promoting food ingredients, advancing the valorization of piperine in functional foods.

The urgent need for new hepatocellular carcinoma (HCC) therapies has driven the exploration of natural product scaffolds. In this context, Icariside II (ICA-II) presents a valuable starting point for structural optimization. Applying bioisosterism and pharmacophore hybridization, we synthesized a series of 26 nitrogen heterocycle-modified ICA-II derivatives. Subsequent structure-activity relationship (SAR) profiling highlighted compound 3b as the most potent analog. This molecule exhibited robust anti-proliferative activity across multiple HCC cell lines (HepG2, Hep3B, and Huh7), surpassing both the parent ICA-II and the positive control Sorafenib. To uncover its primary target, we integrated network pharmacology with molecular docking and dynamics simulations. This computational prediction was further supported by cellular thermal shift assays and EGFR enzymatic inhibition studies, suggesting EGFR as a potential target of 3b. Compound 3b suppressed EGFR phosphorylation and inhibited the downstream PI3K/Akt signaling pathway. This blockade precipitates a profound intracellular accumulation of reactive oxygen species ultimately driving the cancer cells into caspase-dependent apoptosis. Animal studies using a Huh7 xenograft model further corroborated its therapeutic potential. Compound 3b administration markedly suppressed tumor growth without significant body weight loss or obvious histopathological abnormalities in major organs under the tested conditions, notably outperforming the clinically approved agent Icaritin. Preliminary pharmacokinetic studies further demonstrated that 3b achieved robust systemic exposure and a prolonged retention time following intravenous administration. Overall, our work provides a successful structural optimization rationale for ICA-II and advances 3b as a safe, EGFR-modulating lead compound for HCC intervention.

Cynaropicrin is a guaianolide-type sesquiterpene lactone primarily derived from plants within the Asteraceae family. Structurally, it features a distinctive 5-7-5 tricyclic framework, four exocyclic double bonds, and two hydroxyl groups. Due to its broad pharmacological activities, cynaropicrin garnered considerable scientific interest, with well-established protocols for its extraction and synthesis. Extensive studies have demonstrated that cynaropicrin exhibits remarkable antitumor activities by effectively inhibiting the proliferation of various malignancies, such as lung cancer, melanoma, and breast cancer. The underlying mechanisms are primarily associated with the regulation of cell cycle progression and apoptosis-related pathways. Moreover, cynaropicrin significantly enhances the efficacy of chemotherapeutic agents including temozolomide, cisplatin, and docetaxel, suggesting promising potential for combination therapy. Beyond its antitumor properties, cynaropicrin exhibits a broad spectrum of bioactivities, including antioxidant, antiviral, antiparasitic, and immunomodulatory effects, primarily via the modulation of signaling pathways like NF-κB. Structure-activity relationship (SAR) studies further indicate that the side chain, hydrophilicity of the OH-3 and OH-19 substituents, and the C17-C18 exo-olefin structure critically influence its NF-κB inhibitory activity. This review comprehensively synthesizes current knowledge regarding the mechanisms of action and SAR insight of cynaropicrin across various pathophysiological processes, evaluates its prospects for clinical application, and aims to provide a foundational framework to guide future research and development efforts involving this compound.

In this study, the chemical constituents of the branches and leaves of Brucea javanica were investigated using an integrated approach combining molecular networking, the Moldiscovery strategy, and traditional phytochemical techniques. This strategy led to the isolation and characterization of nine previously undescribed triterpenoids, named brujavaneos A-I (1-9). Their structures were unambiguously established through extensive spectroscopic analysis and electronic circular dichroism calculations. Furthermore, the anti-inflammatory activities of these isolates were evaluated. Several compounds exhibited moderate activity, with their IC50 values reliably predicted via InflamNat, a machine-learning-based tool. Notably, compound 4 demonstrated significant inhibition of nitric oxide production in LPS-stimulated RAW264.7 macrophages, with an IC50 value of 15.03 ± 0.03 μM.

The psoriasis is a prevalent chronic inflammatory skin disorder, in which the inflammatory reaction plays an essential role. The genus Daphne contains numerous secondary metabolites with anti-inflammatory activity, which makes it possible to isolate the compounds with anti-inflammatory activity targeted from Daphne Retusa Hemsl. Herein, daphusanes A-P (1-16), sixteen unprecedented guaiane-type sesquiterpenes, along with eighteen known analogues (17-34) were targeting isolated from D. Retusa via molecular networking. The structures of 1-16 were established through NMR spectroscopy analysis, NMR and ECD calculations and single-crystal X-ray diffraction. All the isolated compounds were tested for their anti-inflammatory activity of the compounds against RAW264.7. The preliminary structure-activity relationships of the sesquiterpenoids for the anti-inflammatory activity were summarized. Importantly, 5 showed significant anti-psoriasis activity in imiquimod-induced psoriasis murine model. The metabolomics and Western blot results showed that 5 achieved anti-psoriasis effects by affecting steroid hormone biosynthesis which then affect the RORγT signaling pathway.

Phytochemical investigation on the roots of Stellera chamaejasme Linn. Led to the isolation of twelve undescribed guaiane-type sesquiterpenoids (1-12). Their chemical structures were determined by a comprehensive analysis of spectroscopic data. The absolute configuration was elucidated by comparing experimental ECD. The acetylcholinesterase (AChE) inhibitory activities of isolated compounds were evaluated. Among these, compounds 3 and 9 exhibited comparable inhibitory activity, with IC50 values of 2.81 ± 0.37 μM and 3.22 ± 0.21 μM, even more than Donepezil. Additionally, the molecular modeling studies revealed the structural basis for the significant inhibitory activity of compounds 3 and 9.

Seven undescribed quassinoids (1-7) and one known quassinoid (8) were obtained from Brucea mollis by molecular networking integrated with DeepSAT and SIRIUS. Compound 1 is the first alkaloid-quassinoid adduct featuring a novel 6/5/5/6/6/6-fused hexacyclic skeleton. Compounds 2 and 3 are unprecedented skeleton C19 quassinoids. Biosynthetic pathways were proposed, which guided the successful biomimetic synthesis of compound 1, affording an optimized yield of 23.2%. Byproduct 1a with a rare 5/6/5/6/6/6 core was also formed.

Persicaria tinctoria, a commercially significant cultivated plant, represents the principal source of natural indigo dye and produces the pharmaceutically active metabolite indirubin. Herein, four undescribed anthranilic acid- based alkaloids, (+/-)-persicaritinctories A-C (1a/1b, 3 and 4) and six undescribed natural products (5-9 and 11), along with four known compounds (2, 10, 12 and 13), have been isolated from the leaves of Persicaria tinctoria under the guidance of HSQC-based DeepSAT technology. We elucidated these structures through comprehensive spectroscopic analyses, computer-assisted structure elucidation (CASE) with density functional theory (DFT) calculations, single-crystal X-ray diffraction analysis, and comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Furthermore, the plausible biosynthetic pathways for compounds 1a/1b and 3 were proposed. The isolated alkaloids were subsequently evaluated for their neuroprotective effects and acetylcholinesterase inhibitory activities. These findings not only enhance our understanding of the neuroprotective activity of anthranilic acid alkaloids but also highlight the potential of P. tinctoria for further pharmacological investigation and application development.

Traditional Chinese Medicine (TCM) offers distinct advantages in the treatment of tumors, since it serves dually as both a medicinal treatment and a dietary therapy. Elephantopus scaber (E. scaber), with a plethora of folk medicinal usage for treating pneumonia and hepatitis, contains sesquiterpene lactone (SL) as the primary component for therapeutic efficacy. The orphan drug ACT001, as the SL derivative, has been used in the treatment of glioma, which demonstrated significant potential for the development of such compounds. In this work, two series of plant-derived SL derivatives were synthesized and their efficacy against malignant glioma (MG) was evaluated. Among them, compound 1e exhibited the most potent inhibitory effects with IC50 values of 3.95 and 3.43 μM against U87 and T98G cells, respectively. Preliminary mechanism investigations suggested that 1e induced the cell-cycle arrest at S phase and inhibited the tube formation to the anti-angiogenesis. Meanwhile, 1e enhanced E-cadherin protein level while decreased the levels of Vimentin, MMP-2 and MMP9, thereby suppressing MG cells migration and invasion. Furthermore, the orthotopic glioma model using live animal fluorescence imaging demonstrated the therapeutic effect of 1e on MG in vivo and pharmacokinetic studies indicated 1e with a favorable pharmacokinetic profile. Moreover, we performed competitive activity-based protein profiling (ABPP) to explore the potential targets of SL derivatives in U87 cells, providing a basis for the follow-up studies of MG.