Toosendanin, a novel potent vacuolar-type H+-translocating ATPase inhibitor, sensitizes cancer cells to chemotherapy by blocking protective autophagy.

Macroautophagy/autophagy is the process of self-digestion through the lysosomes; it disassembles unnecessary or dysfunctional long-lived proteins and damaged organelles for the recycling of biomacromolecules. Unfortunately, cancer cells can hijack this mechanism to survive under metabolic stress or develop drug resistance during chemotherapy. Increasing evidence indicates that the combination of autophagy inhibition and chemotherapy is a promising cancer treatment strategy. However, effective autophagy inhibitors with satisfied potency, bioavailability, and clearly-defined drug targets are still rare. Here, we report the identification of a potent autophagy inhibitor toosendanin which can effectively block autophagosome maturation, causing the accumulation of autophagy substrates in multiple cancer cells. Toosendanin did not inhibit the fusion process between autophagosome and lysosome but elevated lysosomal pH and impaired lysosomal enzymes activity. Using rat liver lysosome fraction and purified yeast V-ATPase, we found that toosendanin directly inhibited V-ATPase activity. By applying cellular thermal shift assay (CETSA), immunoprecipitation-coupled LC-MS/MS analysis, and biotin-toosendanin pull-down assay, we confirmed the direct binding between toosendanin and V-ATPase. Furthermore, toosendanin blocked chemotherapy-induced protective autophagy in cultured cancer cells and xenograft tumor tissues to significantly enhance anti-cancer activity. These results suggest that toosendanin has the potential to be developed into an anti-cancer drug by blocking chemotherapy-induced protective autophagy.

Discovery of Three New Monoterpenoid Indole Alkaloids from the Leaves of Gardneria multiflora and Their Vasorelaxant and AChE Inhibitory Activities.

Three novel monoterpenoid indole alkaloids gardflorine A (1), gardflorine B (2), and gardflorine C (3) were isolated from the leaves of Gardneria multiflora. Their structures, including absolute configurations, were established on the basis of spectroscopic methods (MS, UV, IR, 1D and 2D NMR) and circular dichroism experiments. All the compounds were evaluated for their vasorelaxant and acetylcholinesterase (AChE) inhibitory activities. Compound 1 exhibited potent vasorelaxant activity, with an EC50 value of 8.7 µM, and compounds 2 and 3 showed moderate acetylcholinesterase (AChE) inhibitory activities, with IC50 values of 26.8 and 29.2 µM, respectively.

Targeting the NLRP3 inflammasome as new therapeutic avenue for inflammatory bowel disease.

The incidence and prevalence of inflammatory bowel disease (IBD) are increasing worldwide. Current approved medication for IBD treatment in the clinic mainly includes corticosteroids and neutralization antibodies to pro-inflammatory cytokines. However, drug resistance and severe side effect hinder long-term efficacy of these agents. The NOD-like receptor family pyrin domain containing protein 3 (NLRP3) is exclusively expressed in several inflammatory and autoimmune diseases. Excessive expression, aberrant activation, polymorphism, and gain-of-function mutation of the NLRP3 inflammasome contribute to IBD pathogenesis. In this article, we summarize the regulatory factors to NLRP3, and review recently developed NLRP3 inhibitors and their preclinical and clinical applications in treating inflammatory and autoimmune diseases. We present our views on the therapeutic potential of NLRP3 inhibitors as emerging therapeutic avenue for IBD.

Discovery of fused bicyclic derivatives of 1H-pyrrolo[1,2-c]imidazol-1-one as VDR signaling regulators.

The modulation of VDR signaling is important in regulating tumor-related signal transduction and protecting from microorganismal infection. In this study we discovered by luciferase reporter assay that several fused bicyclic derivatives of 1H-pyrrolo[1,2-c]imidazol-1-one with the assistance of calcitriol result in up to three-fold increases of VDR promoter activity. Preliminary SAR results from 20 compounds disclose that ideal VDR signaling regulators of these compounds are built up by the optimal combination of multiple factors. Western blot analysis indicates that compounds of ZD-3, ZD-4 and ZD-5 not only significantly upregulate p62 and LC3-II but also elevate the ratio of LC3-II/LC3-I, which possibly leads to activated autophagy. All of five compounds also significantly downregulate p65 and upregulate p-p65 and ZD-3 is the most active one to NF-κB signaling, suggesting a possible induction of apoptosis through the regulation of NF-κB signal transduction mediated by VDR signaling. Compounds of ZD-3, ZD-4 and ZD-5 significantly counteract the interference by VDR shRNA, in which ZD-3 gets the highest compensation of VDR expression and the highest ratio of LC3-II/LC3-I, indicating that ZD-3 very likely activates VDR-mediated autophagy. Taken together, these 1H-pyrrolo[1,2-c]imidazol-1-one derivatives can modulate VDR signaling, possibly resulting in the regulation of some signal pathways to induce autophagy and apoptosis.

Iron(III)-Selective Chelation-Enhanced Fluorescence Sensing for In Vivo Imaging Applications.

A "turn-on" pattern Fe(3+) -selective fluorescent sensor was synthesized and characterized that showed high fluorescence discrimination of Fe(3+) over Fe(2+) and other tested ions. With a 62-fold fluorescence enhancement towards Fe(3+) , the probe was employed to detect Fe(3+) in vivo in HeLa cells and Caenorhabditis elegans, and it was also successfully used to elucidate Fe(3+) enrichment and exchange infected by innexin3 (Inx3) in hemichannel-closed Sf9 cells.

[Chemical constituents from the aerial part of Echinacea purpurea].

OBJECTIVE: To study the chemical constituents of the aerial part of Echinacea purpurea. METHODS: The compounds were separated and purified by repeatedly silica gel, ODS, D101 macroporous resin, MCI, Sephadex LH-20 column chromatography and recrystallization. Their structures were elucidated on the basis of physiochemical properties and spectral analysis. RESULTS: Sixteen compounds were isolated and identified as (2S)-1-O-octacosanoyl glycerol (1), (5R,6S)-6-hydroxy-6-((E)-3-hydroxybut-1-enyl)-1,1, 5-trimethylcyclohexanone (2), (3S, 6E, 10R)-3, 10, 11-trihydroxy-3, 7, 11-trimethyl-dodeca-1, 6-diene (3), negunfurol (4), schensianol A (5), ent-4 (15) -eudesmene-1beta, 6alpha-diol (6), (E) -5-hydroxy-N-isobutylpentadec-2-enamide (7), syringaresinol (8), quercetin (9), ethyl laurate (10), ethyl caffeate (11), ferulic acid (12), alpha-spinasterol (13), stigmasterol (14), beta-daucosterol (15), octacosanoic acid (16). CONCLUSION: Compound 1 - 5 are isolated from the Asteraceae for the first time, compound 6 ,7, 9, 10, 12 are isolated from genus of Echinacea for the first time, compound 15, 16 are isolated from this plant for the first time.