Pyruvate carboxylase (PC) replenishes tricarboxylic acid cycle intermediates, driving cancer metabolic reprogramming. To improve the metabolic stability of erianin, a potent PC inhibitor from Dendrobium chrysotoxum Lindl, we designed and synthesized 55 derivatives, culminating in the identification of CIB-Q22, which exhibited potent PC inhibition (IC50 = 1.74 nM) and suppressed HCC cell viability (IC50 = 25.18 nM), comparable to erianin. Notably, CIB-Q22 demonstrated significantly improved in vivo stability, with a half-life (T1/2 = 1.21 h) much longer than erianin (T1/2 ∼ 0.1 h). Mechanistically, CIB-Q22 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. In vivo, CIB-Q22 exhibited comparable antitumor efficacy but improved safety compared to sorafenib. With its potent PC inhibition and favorable drug-like properties, CIB-Q22 represented a promising therapeutic candidate for HCC treatment.

Dendrobium nobile Lindl. is a major source of "Shi-Hu" listed in the 'Chinese Pharmacopoeia'. D. nobile has been cultivated in large areas. A comprehensive investigation of the chemical constituents of D. nobile is pivotal for production, exploitation, and quality control. A detailed examination on the chemical components led to the identification of 81 compounds, including 23 undescribed ones. The undescribed compounds consist of 16 sesquiterpenoids (1-14, 16, and 17), one alkaloid (15), one phenanthrene (18), and five bibenzyls (19-23). Compound 1 is a structurally unique sesquiterpenoid, which is possibly biosynthesized from a copacamphane-type sesquiterpene. Compounds 16 and 17 are the first muurolane-type sesquiterpenoids with demethylation of C-15. Compounds 14, 19, and 59 significantly inhibited the collagen formation in TGF-β induced NIH 3T3 cells, with IC50 values of 5.83, 7.48, and 7.43 μM, respectively, suggesting their potential to inhibit pulmonary fibrosis.

Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations are associated with poor prognosis and poor response to standard therapeutic regimens in patients with nonsmall cell lung cancer (NSCLC). Identification of novel synthetic lethal partners in oncogenic KRAS is an alternative therapeutic strategy for KRAS-mutant malignancies. After high-throughput screening against a preclinical/clinical compound library, embelin, a known X-linked inhibitor of apoptosis protein (XIAP) inhibitor, specifically inhibits the catalytic activity and phosphorylation of Src homology domain 2 containing tyrosine phosphatase 2 (SHP2) in KRAS-mutant NSCLC cells. Pharmacological inhibition and genetic knockdown of XIAP and SHP2 induce synthetic lethality in KRAS-mutated NSCLC cells and xenograft animal models. Mechanistically, dual inhibition of XIAP and SHP2 by embelin lessens the proliferation and metastasis, activates senescence and endogenous apoptosis, inhibits cancer-related RAS/mitogen-activated protein kinase (MAPK), phosphoinositide-3-kinase (PI3K)/AKT, Janus kinase/signal transducers and activators of transcription (JAK/STAT), wingless-related integration site (Wnt), and nuclear factor kappa B (NF-κB) signaling pathways, and overcomes compensatory feedback in the MAPK signals through the modulation of mitogen-inducible gene-6 (MIG-6) and SPROUTY2 (SPRY2). Collectively, SHP2 and XIAP are potential synthetic lethal partners, and embelin warrants further development as a novel therapeutic option for alleviating KRAS-mutant NSCLC by cotargeting SHP2 and XIAP.

New therapeutic approaches are essential in the fight against breast cancer, which remains one of the top causes of mortality globally. Innovative and efficient methods of treating and preventing cancer has become expedient since its incidence rates are rising globally. Combining herbal extracts and chemotherapy have drawn a lot of attention in recent times as a cutting-edge cancer prevention approach. The wild parasitic plant Loranthus micranthus is extensively distributed throughout the world and is well-known for its therapeutic uses. Previous preclinical investigations indicated that the leaves and stem extracts of L. micranthus had the potential to suppress breast cancer. Investigating the anticancer effects of L. micranthus extracts through network pharmacology analysis, in vitro and in vivo experiments is the goal of the current study. Network pharmacology analysis revealed 207 targets and 30 bioactive phytoconstituents of L. micranthus associated with the metabolism of breast cancer. L. micranthus controlled the metabolism of tryptophan and nitrogen in breast cancer, according to KEGG analysis and in silico models. The results of the experiment showed that L. micranthus significantly reduced the synthesis of kynurenine in interferon-γ (IFN-γ)-stimulated breast cancer cells, downregulated important proteins involved in tryptophan catabolism, and produced no cytotoxic effects in human breast cancer cells (MCF 7 and MDA-MB 231) at the administered doses. The viability of T cells co-cultured with IFN-γ-treated breast cancer cells was also markedly enhanced by L. micranthus pre-treatment. The in vivo investigation showed a similar outcome, with L. micranthus treatment suppressing the inflammatory response, IDO activity/expression, lowering kynurenine levels, blocking CTLA-4 immune checkpoint and finally increasing the CD4+ T cell population in rats with DMBA-induced breast cancer.

Endocrine disturbance induces the incidence of polycystic ovary syndrome (PCOS) in women. Inhibiting adenylate cyclase (AC), which participates in the function of disturbed endocrine factors such as follicle-stimulating hormone (FSH), could be a novel therapeutic strategy to treat PCOS. In this study, a novel AC activity assay was established and used for high-throughput screening of a Food and Drug Administration (FDA)-approved drug library. Consequently, aspirin effectively inhibited the activity of AC in vitro and reduced estrogen biosynthesis by inhibiting the expression of aromatase in human granulosa cells, a key enzyme that catalyzes the conversion of androgens into estrogens. Aspirin irreversibly inhibited the activity of AC subtypes and covalently bound to recombinant AC by binding to Lys350 and Lys426, which correspond to Lys938 and Lys1014 of full-length AC2. Mechanistically, aspirin inhibited cAMP response element-binding protein-mediated aromatase expression by the AKT/mammalian target of rapamycin (mTOR) signaling pathway. Furthermore, aspirin effectively alleviated PCOS symptoms in mouse models by decreasing androgen levels, lowering insulin levels, and reducing cystic follicles. These results revealed that aspirin is a potent inhibitor of ACs and a previously unrecognized role of ACs in estrogen biosynthesis. Therefore, aspirin warrants further investigation as a novel therapeutic agent in PCOS.

The stems of Dendrobium chrysotoxum Lindl are used as medicinal herbs and for nutraceutical beverages and functional food products. In order to obtain an in-depth understanding of the bioactive compounds in the stems of D. chrysotoxum Lindl, an exhaustive chemical examination was carried out. A total of 20 undescribed compounds (1-20) along with 40 known ones (21-60) were isolated. The structures of 1-20 were elucidated based on spectroscopic analysis and theoretical quantum calculations. The unreported compounds consist of 7 spirophenanthrenes (1-7), 4 diphenanthrenes (8-11), 1 9,10-dihydrophenanthrene derivative (12), 1 phenanthropyran (13), 2 monophenanthrenes (14 and 15), 1 dimeric bibenzyl (16), and 3 bibenzyls (17-20). Six compounds 1b, 3b, 4a, 4b, 8, and 16 exhibited remarkable cytotoxic activity against HCCLM3 cancer cells with IC50 values of 1.68 μM-6.60 μM. Compounds 10, 15, and fimbriadimerbibenzyl D displayed significant anti-pulmonary fibrosis with IC50 values of 1.82 ± 0.02 μM to 3.68 ± 0.03 μM.

Src homology-2-containing protein tyrosine phosphatase 2 (SHP2) plays a dual role in cancer initiation and progression. Identifying signals that modulate the function of SHP2 can improve current therapeutic approaches for IFN-α/β in HCC. We showed that cAMP-dependent PKA suppresses IFN-α/β-induced JAK/STAT signaling by increasing the phosphatase activity of SHP2, promoting the dissociation of SHP2 from the receptor for activated C-kinase 1 (RACK1) and binding to STAT1. Additionally, cAMP-degrading phosphodiesterase 4D (PDE4D) physically interacts with RACK1 to regulate PKA-mediated SHP2 activity and STAT1 phosphorylation. IFN-α activates PKA by inducing the expression of cyclooxygenase 2 (COX2) and the production of prostaglandin E2 (PGE2), which in turn stimulates the binding of SHP2 to IFNAR2 via RACK1. A COX inhibitor aspirin potently increases the antitumor effects of IFN-α in the suppression of HCC cell proliferation in vivo. Higher expression of COX2 and phosphorylated STAT3 is associated with poor development and prognosis in HCC patients by analyzing human HCC clinical samples. These observations suggest that a fundamental PKA/SHP2-dependent negative feedback loop acts on IFN signaling, and inhibition of this signaling by the selective COX2 inhibitors may enhance the clinical efficacy of type I IFNs in treating HCC.

Two novel R-gamma ' type lignans, notolignan A (2) and notolignan B (3) were isolated from the 80 % EtOH extract of cultivated Notopterygium incisum along with one known analogue (1) by normal and reverse phase column chromatography. The structures of the undescribed compounds were characterized by extensive spectroscopic data. This study aimed to evaluate the potential protective effects of a novel compound on acute lung injury (ALI) in mice via the NLRP3 inflammasome. The model of ALI was established with lipopolysaccharide (LPS) in C57BL/6 mice by intraperitoneal injection. LPS was also used to stimulate RAW264.7 cells that detected the antiinflammatory activity of compound 2. The NLRP3 inflammasome was activated by LPS + Nig in iBMDMs. Our data noted that compound 2 could effectively inhibit the NO production with safe concentration and which effect was superior to positive control (30, 40 and 50 mu M). Furthermore, compound 2 dramatically suppresses the activation of NLRP3 inflammasome and production of IL-1R, blocking NEK7-NLRP3 and NLRP3-ASC interactions. The DFT calculations, docking, and MDs have demonstrated that compound 2 interacts with NLRP3 to form a stable complex. In addition, pre-treated with compound 2 could obviously alleviated the lung pathological symptoms in ALI mice, and significantly down regulate the expression of NLRP3 and IL-1R in lung tissues. These findings suggest that the isolated compounds may serve as potential anti-inflammatory agents or lead compounds for drug development.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease for which few drugs are available in clinical practice. Here, we identified novel capsaicin analogs by combining in-house chemical library screening and further structural optimization. (E)-1-(3,4-dihydroxyphenyl)-7-phenylhept-1-en-3-one (Compound 14) was found to be the most potent in inhibiting TGF-beta-induced collagen accumulation, proliferation and migration in fibroblast cells. Furthermore, compound 14 (IC50 = 0.51 +/- 0.06 mu M) showed over 100-fold increasing antifibrotic activity compared to capsaicin (IC50 = 53.71 +/- 4.78 mu M). Notably, compound 14 could target TRPV1, thereby affecting the expression of the fibrosis markers Collagen I and alpha-SMA by inhibiting the TGF-beta/Smads and MAPK pathways to exert antifibrotic activity in vitro. Compound 14 significantly inhibited collagen deposition in lung tissues, ameliorated alveolar structures, and increased survival rates in mice with bleomycin-induced pulmonary fibrosis. In addition, compound 14 possessed lower cytotoxicity (compared to nitedanib) and no toxicity in mice. Overall, compound 14 promise as a potential drug candidate for the treatment of IPF.

Interferon alpha (IFNα) leads to therapeutic effects on various tumors, especially renal cell cancer (RCC), by directly protecting against tumors cell proliferation or indirectly inducing an anti-tumor immune response. However, new combination therapies are needed to enhance the efficacy of IFNα and reduce its adverse effects during long-term treatment. In this study, we found that the anti-proliferative effects of IFNα on RCC cells in vitro and in vivo were greater after the allosteric inhibition of SHP2 by SHP099 than after treatment with enzymatic inhibitors of SHP2. SHP099 increased IFNα-induced pro-caspase-1 expression in RCC cells, activated the NLRP3 inflammasome, and induced pyroptosis. Mechanistically, SHP099 not only increased the expression of NLRP3 inflammasome components via the NF-κB signaling pathway, but also further activated the NLRP3 inflammasome by regulating mitochondrial homeostasis through ANT1-mediated reactive oxygen species modulation. Allosteric inhibition of SHP2 by SHP099 also potently enhanced the anti-tumor immunity induced by IFNα by modulating T cell proliferation and infiltration in vitro and in vivo. These results reveal the new function of SHP2 in NLRP3 inflammasome activation and pyroptosis in RCC and provide a basis for further investigating the combination of allosteric SHP2 inhibitors with IFNα in cancer immunotherapy.

Decarboxylative C(sp2)-heteroatom cross-coupling reactions hold extraordinary potential for the sustainable preparation of biologically active scaffolds. Herein, we report a copper sulfate/1,10-phenathroline catalytic system for the decarboxylative intramolecular C(sp2)-O, C(sp2)-S, and C(sp2)-N coupling reactions leading to the construction of a series of benzo[b]furans, benzo[b]thiophenes, and indole derivatives from the corresponding coumarins, thiocoumarins, or quinolones, respectively. Our mechanistic study based on benzo[b]furan formation suggests a three-step process of the transformations, which consists of (i) base-mediated hydrolytic ring opening of coumarin, (ii) copper-oxygen co-initiated radical decarboxylation, and (iii) copper-catalyzed C-heteroatom cross coupling. Application of this method in the total synthesis of egonol, a bioactive natural product, was demonstrated successfully, with an overall yield of 51.7%.

ETHNOPHARMACOLOGICAL RELEVANCE:Leonurus japonicus Houtt (L. japonicus, Chinese motherwort), known as Yi Mu Cao which means "good for women", has long been widely used in China and other Asian countries to alleviate gynecological disorders, often characterized by estrogen dysregulation. It has been used for the treatment of polycystic ovary syndrome (PCOS), a common endocrine disorder in women but the underlying mechanism remains unknown.AIM OF THE STUDY:The present study was designed to investigate the effect and mechanism of flavonoid luteolin and its analog luteolin-7-methylether contained in L. japonicus on aromatase, a rate-limiting enzyme that catalyzes the conversion of androgens to estrogens and a drug target to induce ovulation in PCOS patients.MATERIALS AND METHODS:Estrogen biosynthesis in human ovarian granulosa cells was examined using ELISA. Western blots were used to explore the signaling pathways in the regulation of aromatase expression. Transcriptomic analysis was conducted to elucidate the potential mechanisms of action of compounds. Finally, animal models were used to assess the therapeutic potential of these compounds in PCOS.RESULTS:Luteolin potently inhibited estrogen biosynthesis in human ovarian granulosa cells stimulated by follicle-stimulating hormone. This effect was achieved by decreasing cAMP response element-binding protein (CREB)-mediated expression of aromatase. Mechanistically, luteolin and luteolin-7-methylether targeted tumor progression locus 2 (TPL2) to suppress mitogen-activated protein kinase 3/6 (MKK3/6)-p38 MAPK-CREB pathway signaling. Transcriptional analysis showed that these compounds regulated the expression of different genes, with the MAPK signaling pathway being the most significantly affected. Furthermore, luteolin and luteolin-7-methylether effectively alleviated the symptoms of PCOS in mice.CONCLUSIONS:This study demonstrates a previously unrecognized role of TPL2 in estrogen biosynthesis and suggests that luteolin and luteolin-7-methylether have potential as novel therapeutic agents for the treatment of PCOS. The results provide a foundation for further development of these compounds as effective and safe therapies for women with PCOS.

Myeloproliferative neoplasms (MPNs), including polycythemia vera, essential thrombocytosis, and primary myelofibrosis, are clonal hematopoietic neoplasms driven by mutationally activated signaling by the JAK2 tyrosine kinase. Although JAK2 inhibitors can improve MPN patients' quality of life, they do not induce complete remission as disease-driving cells persistently survive therapy. ERK activation has been highlighted as contributing to JAK2 inhibitor persistent cell survival. As ERK is a component of signaling by activated RAS proteins and by JAK2 activation, we sought to inhibit RAS activation to enhance responses to JAK2 inhibition in preclinical MPN models. We found the SHP2 inhibitor RMC-4550 significantly enhanced growth inhibition of MPN cell lines in combination with the JAK2 inhibitor ruxolitinib, effectively preventing ruxolitinib persistent growth, and the growth and viability of established ruxolitinib persistent cells remained sensitive to SHP2 inhibition. Both SHP2 and JAK2 inhibition diminished cellular RAS-GTP levels, and their concomitant inhibition enhanced ERK inactivation and increased apoptosis. Inhibition of SHP2 inhibited the neoplastic growth of MPN patient hematopoietic progenitor cells and exhibited synergy with ruxolitinib. RMC-4550 antagonized MPN phenotypes and increased survival of an MPN mouse model driven by MPL-W515L. The combination of RMC-4550 and ruxolitinib, which was safe and tolerated in healthy mice, further inhibited disease compared to ruxolitinib monotherapy, including extending survival. Given SHP2 inhibitors are undergoing clinical evaluation in patients with solid tumors, our preclinical findings suggest that SHP2 is a candidate therapeutic target with potential for rapid translation to clinical assessment to improve current targeted therapies for MPN patients.

Transactivation response (TAR) RNA-binding protein 2 (TRBP) plays a critical role in microRNA (miRNA) biosynthesis, with aberrant expression linked to various cancers. Previously, we identified CIB-3b, a phenyloxazole derivative that disrupts the TRBP-Dicer interaction in hepatocellular carcinoma (HCC). In this study, we optimized this scaffold and substituent, leading to the discovery of CIB-L43, a 2-phenylthiazole-5-carboxylic acid derivative with nanomolar inhibitory activity (EC50 = 0.66 nM). CIB-L43 demonstrated superior TRBP binding affinity (KD = 4.78 nM) and enhanced disruption of TRBP-Dicer interactions (IC50 = 2.34 μM). Mechanistically, CIB-L43 suppressed oncogenic miR-21 biosynthesis, increasing PTEN and Smad7 expression and inhibiting AKT and TGF-β signaling, thereby reducing HCC cell proliferation and migration. In vivo, CIB-L43 exhibited favorable pharmacokinetics, including 53.9% oral bioavailability, and comparable antitumor efficacy to first-line anticancer drug, sorafenib, with lower toxicity. CIB-L43 emerges as a promising HCC treatment candidate with potent TRBP inhibition and favorable drug-like properties.

Caspase-1 plays a central role in innate immunity, as its activation by inflammasomes induces the production of proinflammatory cytokines and pyroptosis. However, specific inhibition of the enzymatic activity of this protease is not effective in suppressing inflammation, owing to its enzyme-independent function. Herein, we identified a cyclohexenyl isothiocyanate compound (CIB-1476) that potently inhibited caspase-1 activity and suppressed the assembly and activation of the NLRP3 inflammasome and gasdermin-D-mediated pyroptosis. Mechanistically, CIB-1476 directly targeted pro-caspase-1 as an irreversible covalent inhibitor by binding to Cys285 and Cys397, resulting in more durable anti-inflammatory effects in the suppression of enzyme-dependent IL-1β production and enzyme-independent nuclear factor κB activation. Chemoproteomic profiling demonstrated the engagement of CIB-1476 with caspase-1. CIB-1476 showed potent therapeutic effects by suppressing inflammasome activation in mice, which was abolished in Casp1 –/– mice. These results warrant further development of CIB-1476 along with its analogues as a novel strategy for caspase-1 inhibitors.

Dysfunction of calcium channels is involved in the development and progression of some cancers. However, it remains unclear the role of calcium channel inhibitors in tumor immunomodulation. Here, calcium channel blocker lacidipine is identified to potently inhibit the enzymatic activity and expression of indoleamine 2,3-dioxygenase 1 (IDO1), a rate-limiting enzyme in tryptophan metabolism. Lacidipine activates effector T cells and incapacitates regulatory T cells (Tregs) to augment the anti-tumor effect of chemotherapeutic agents in breast cancer by converting immunologically "cold" into "hot" tumors. Mechanistically, lacidipine targets calcium channels (CaV1.2/1.3) to inhibit Pyk2-JAK1-calmodulin complex-mediated IDO1 transcription suppression, which suppresses the kynurenine pathway and maintains the total nicotinamide adenine dinucleotide (NAD) pool by regulating NAD biosynthesis. These results reveal a new function of calcium channels in IDO1-mediated tryptophan metabolism in tumor immunity and warrant further development of lacidipine for the metabolic immunotherapy in breast cancer.

The fragrant flowers of Rosa hugonis Hemsl. Contain abundant valuable rose oil and carotenoids. However, phytochemical investigation of this resource rich in phenolics with neuroprotective activity in vitro has been rarely reported. Purification of the 70% ethanol extracts from the flowers of R. hugonis by various chromatographic methods resulted in the isolation and characterization of five undescribed acylated flavonoid glycosides (Hugonisflavonoid A-E) together with forty known phenolics. The chemical structures of the undescribed compounds were elucidated by extensive analysis of their spectroscopic data and chemical methods. All the isolates were found from R. hugonis for the first time and evaluated for their neuroprotective effects on 6-OHDA induced injury in PC12 cells. Seventeen compounds displayed remarkable protective effects at concentrations of 10 μM. Hugonisflavonoid E can reduce excessive reactive oxygen species and up-regulate mRNA expression levels of superoxide dismutase 1 and catalase. Additionally, hugonisflavonoid E activated the phosphorylated proteins such as PDK1, Akt and GSk-3β. These findings suggested that R. hugonis could be a potential source for neuroprotective agents.

Three undescribed dammarane-type triterpene saponins, 20(S)-sanchirhinoside A7-A9 (1-3), together with seventeen known ones, were isolated from the roots of Panax notoginseng (Burk.) F. H. Chen. The chemical structures of the new compounds were determined by HR-MS and NMR experiments along with chemical methods. To the best of our knowledge, compound 1 was the firstly reported fucose-containing triterpene saponin from plants in the genus of Panax. Moreover, the in vitro neuroprotective effects of the isolated com-pounds were evaluated. Compounds 11-12 displayed remarkable protective effects against PC12 cells injured by 6-hydroxydopamine.

The mechanism of the selective protection of L-lysine mediated by β-cyclodextrin (β-CD) was investigated by preliminary experiments, including the reaction efficiency influenced by different reaction conditions, and the existence of (1a·CD)' and 1a·CD·2a was evidenced by ESI-MS and 2D Rotating Frame Overhauser Effect Spectroscopy (ROESY) analysis. The results indicated that the formation of (1a·CD)' is critical for the product selectivity and the further formation of the ternary complex 1·CD·2 is responsible for the reaction efficiency. Thus, the yields and selectivity were significantly influenced by the structure, size and reactivity of the reactants. During the mechanistic investigations, we realized that the formation of the product and the β-CD complex at the final stage of the reaction would cause difficulty in product purification by a previously reported homogeneous method. In light of this understanding, an efficient and practical protocol for selective protection of L-lys based on a heterogeneous catalyst SiO2@CD was developed. The use of the SiO2 immobilized β-CD catalyst prevented the formation of the "capped" products by controlling the spatial rearrangement of β-CDs on solid supports, which represents a considerable synthetic improvement over the tedious and wasteful organic solvent extraction for product purification.