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The FEBS Journal Sep 2010Diterpenes are a structurally diverse class of molecules common in plants, although they are very rarely found in bacteria. We report the identification in Mycobacterium...
Diterpenes are a structurally diverse class of molecules common in plants, although they are very rarely found in bacteria. We report the identification in Mycobacterium tuberculosis (Mtb) of three diterpenes proposed to promote phagolysosome maturation arrest. MS analysis reveals that these diterpenes are novel compounds not previously identified in other organisms. The diterpene with highest abundance in Mtb has a mass fragmentation pattern identical to edaxadiene, which is produced in vitro from geranylgeranyl diphosphate by the enzymes Rv3377c and Rv3378c. A second diterpene found in Mtb has a similar mass spectrum, and is always observed in the same proportion relative to edaxadiene, indicating that it is a side product of the Rv3378c reaction in vivo. We name this second diterpene olefin edaxadiene B. The least abundant of the three diterpenes in Mtb extracts is tuberculosinol, a dephosphorylated side-product of the edaxadiene pathway intermediate produced by Rv3377c. A frameshift in Rv3377c in Mtb completely eliminates diterpene production, whereas expression of Rv3377c and Rv3378c in the nonpathogenic M. smegmatis is sufficient to produce edaxadiene and edaxadiene B. These studies define the pathway of edaxadiene and edaxadiene B biosynthesis in vivo. Rv3377c and Rv3378c are unique to Mtb and M. bovis, making them candidates for selective therapeutics and diagnostics.
Topics: Diterpenes; Mass Spectrometry; Molecular Conformation; Mycobacterium tuberculosis; Stereoisomerism
PubMed: 20670276
DOI: 10.1111/j.1742-4658.2010.07767.x -
Journal of Traditional Chinese Medicine... Oct 2022To systematically evaluate the anti-inflammatory potential of diterpene lactones from Chuanxinlian () (AP).
OBJECTIVE
To systematically evaluate the anti-inflammatory potential of diterpene lactones from Chuanxinlian () (AP).
METHODS
We firstly adopted zebrafish, a novel and ideal animal model for high-throughput drug screening, to investigate the anti-inflammatory activities of 17 diterpene lactones isolated from AP.
RESULTS
The results showed that most of diterpene lactones displayed significant anti-inflammatory effects in lipopolysaccharide microinjection-, copper sulfate exposure- or tail transection-induced zebrafish inflammation models. Moreover, diterpene lactone 3-deoxy-andrographoside (AP-5) was firstly found to attenuate inflammatory responses, which was closely associated with the myeloid differentiation primary response 88/nuclear factor-kappa B and signal transducer and activator of transcription 3 pathways.
CONCLUSION
Our research sheds light on the inestimable roles of zebrafish in high-throughput drug screening, elucidates the potent inhibitory effects of diterpene lactones against inflammation and indicates that AP-5 may serve as a potential alternative agent for the treatment of inflammatory diseases.
Topics: Andrographis paniculata; Animals; Anti-Inflammatory Agents; Diterpenes; Drug Evaluation, Preclinical; Inflammation; Lactones; Lipopolysaccharides; NF-kappa B; Plant Extracts; Zebrafish
PubMed: 36083482
DOI: 10.19852/j.cnki.jtcm.2022.05.008 -
Biochemistry Nov 2022Fusicoccadiene synthase from the fungus (PaFS) is an assembly-line terpene synthase that catalyzes the first two steps in the biosynthesis of Fusiccocin A, a diterpene...
Fusicoccadiene synthase from the fungus (PaFS) is an assembly-line terpene synthase that catalyzes the first two steps in the biosynthesis of Fusiccocin A, a diterpene glycoside. The C-terminal prenyltransferase domain of PaFS catalyzes the condensation of one molecule of C dimethylallyl diphosphate and three molecules of C isopentenyl diphosphate to form C geranylgeranyl diphosphate, which then transits to the cyclase domain for cyclization to form fusicoccadiene. Previous structural studies of PaFS using electron microscopy (EM) revealed a central octameric prenyltransferase core with eight cyclase domains tethered in random distal positions through flexible 70-residue linkers. However, proximal prenyltransferase-cyclase configurations could be captured by covalent cross-linking and observed by cryo-EM and mass spectrometry. Here, we use cryo-EM to show that proximally configured prenyltransferase-cyclase complexes are observable even in the absence of covalent cross-linking; moreover, such complexes can involve multiple cyclase domains. A conserved basic patch on the prenyltransferase domain comprises the primary touchpoint with the cyclase domain. These results support a model for transient prenyltransferase-cyclase association in which the cyclase domains of PaFS are in facile equilibrium between proximal associated and random distal positions relative to the central prenyltransferase octamer. The results of biophysical measurements using small-angle X-ray scattering, analytical ultracentrifugation, dynamic light scattering, and size-exclusion chromatography in-line with multi-angle light scattering are consistent with this model. This model accordingly provides a framework for understanding substrate transit between the prenyltransferase and cyclase domains as well as the cooperativity observed for geranylgeranyl diphosphate cyclization.
Topics: Dimethylallyltranstransferase; Alkyl and Aryl Transferases; Diterpenes
PubMed: 36227241
DOI: 10.1021/acs.biochem.2c00509 -
Marine Drugs Oct 2021Four new indole-diterpenoids, named penerpenes K-N (-), along with twelve known ones (-), were isolated from the fermentation broth produced by adding L-tryptophan to...
Four new indole-diterpenoids, named penerpenes K-N (-), along with twelve known ones (-), were isolated from the fermentation broth produced by adding L-tryptophan to the culture medium of the marine-derived fungus sp. KFD28. The structures of the new compounds were elucidated extensively by 1D and 2D NMR, HRESIMS data spectroscopic analyses and ECD calculations. Compound represents the second example of paxilline-type indole diterpene bearing a 1,3-dioxepane ring. Three compounds (, and ) were cytotoxic to cancer cell lines, of which compound was the most active and showed cytotoxic activity against the human liver cancer cell line BeL-7402 with an IC value of 5.3 μM. Moreover, six compounds (, , , , , and ) showed antibacterial activities against ATCC 6538 and ATCC 6633.
Topics: Animals; Anti-Bacterial Agents; Antineoplastic Agents; Aquatic Organisms; Cell Line, Tumor; Diterpenes; Humans; Indoles; Magnetic Resonance Spectroscopy; Microbial Sensitivity Tests; Penicillium; Staphylococcus aureus
PubMed: 34822484
DOI: 10.3390/md19110613 -
European Journal of Medicinal Chemistry 2013Penitrems are indole diterpene alkaloids best known for their BK channel inhibition and tremorgenic effects in mammals. In a previous study, penitrems A-F (1-5), their...
Penitrems are indole diterpene alkaloids best known for their BK channel inhibition and tremorgenic effects in mammals. In a previous study, penitrems A-F (1-5), their biosynthetic precursors, paspaline (6) and emindole SB (7), and two brominated penitrem analogs 8 and 9 demonstrated promising in vitro antiproliferative, antimigratory, and anti-invasive effects in the MTT (MCF-7 and MDA-MB-231), wound-healing, and Cultrex BME cell invasion (MDA-MB-231) assays, respectively. The study herein reports the novel ability of penitrem A to suppress total β-catenin levels in MDA-MB-231 mammary cancer cells. Nine new penitrem analogs (10-18) were semisynthetically prepared, in an attempt to identify pharmacophores correlated with BK channel inhibition and tremorgenicity of penitrems and decrease their toxicity. The degree of BK channel inhibition was assessed using the nematode Caenorhabditis elegans, and in vivo tremorgenic EC₅₀ was calculated using CD-1 male mice following an Up-and-Down Procedure (UDP). Although new analogs were generally less active than parent compound 1, some showed no BK channel inhibition or tremorgenicity and retained the ability of penitrem A (1) to suppress total β-catenin levels in MDA-MB-231 cells. Paspaline (6) and emindole SB (7), both lacking BK channel inhibition and tremorgenicity, represent the simplest indole diterpene skeleton that retains the antiproliferative, antimigratory and total β-catenin suppressing effects shown by the more complex penitrem A (1).
Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Caenorhabditis elegans; Cell Line, Tumor; Cell Proliferation; Cell Survival; Diterpenes; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Female; Humans; Indole Alkaloids; MCF-7 Cells; Male; Mice; Molecular Structure; Mycotoxins; Structure-Activity Relationship; Wnt Proteins; beta Catenin
PubMed: 24211635
DOI: 10.1016/j.ejmech.2013.09.045 -
Marine Drugs Dec 2021Five undescribed harziane-type diterpene derivatives, namely harzianol K (), harzianol L (), harzianol M (), harzianol N (), harzianol O (), along with two known...
Five undescribed harziane-type diterpene derivatives, namely harzianol K (), harzianol L (), harzianol M (), harzianol N (), harzianol O (), along with two known compounds, hazianol J () and harzianol A () were isolated from the deep-sea sediment-derived fungus sp. SCSIOW21. The relative configurations were determined by meticulous spectroscopic methods including 1D, 2D NMR spectroscopy, and HR-ESI-MS. The absolute configurations were established by the ECD curve calculations and the X-ray crystallographic analysis. These compounds (, and -) contributed to increasing the diversity of the caged harziane type diterpenes with highly congested skeleton characteristics. Harzianol J () exhibited a weak anti-inflammatory effect with 81.8% NO inhibition at 100 µM.
Topics: Animals; Anti-Inflammatory Agents; Antifungal Agents; Aquatic Organisms; Diterpenes; Nitric Oxide; Structure-Activity Relationship; Trichoderma
PubMed: 34940688
DOI: 10.3390/md19120689 -
Plant Biotechnology Journal Jan 2023Selaginella moellendorffii miltiradiene synthase (SmMDS) is a unique bifunctional diterpene synthase (diTPS) that catalyses the successive cyclization of...
Selaginella moellendorffii miltiradiene synthase (SmMDS) is a unique bifunctional diterpene synthase (diTPS) that catalyses the successive cyclization of (E,E,E)-geranylgeranyl diphosphate (GGPP) via (+)-copalyl diphosphate (CPP) to miltiradiene, which is a crucial precursor of important medicinal compounds, such as triptolide, ecabet sodium and carnosol. Miltiradiene synthetic processes have been studied in monofunctional diTPSs, while the precise mechanism by which active site amino acids determine product simplicity and the experimental evidence for reaction intermediates remain elusive. In addition, how bifunctional diTPSs work compared to monofunctional enzymes is attractive for detailed research. Here, by mutagenesis studies of SmMDS, we confirmed that pimar-15-en-8-yl is an intermediate in miltiradiene synthesis. Moreover, we determined the apo-state and the GGPP-bound state crystal structures of SmMDS. By structure analysis and mutagenesis experiments, possible contributions of key residues both in class I and II active sites were suggested. Based on the structural and functional analyses, we confirmed the copal-15-yl intermediate and unveiled more details of the catalysis process in the SmMDS class I active site. Moreover, the structural and experimental results suggest an internal channel for (+)-CPP produced in the class II active site moving towards the class I active site. Our research is a good example for intermediate identification of diTPSs and provides new insights into the product specificity determinants and intermediate transport, which should greatly facilitate the precise controlled synthesis of various diterpenes.
Topics: Alkyl and Aryl Transferases; Diterpenes
PubMed: 36161753
DOI: 10.1111/pbi.13933 -
Arquivos Brasileiros de Cardiologia Oct 2020
Topics: Animals; Arterial Pressure; Blood Pressure; Diterpenes; Hypertension; Rats
PubMed: 33111869
DOI: 10.36660/abc.20200552 -
Marine Drugs Nov 2023In our chemical investigation into sp. UJNMF0740 derived from mangrove sediment, fourteen indole diterpene analogs, including four new ones, are purified by multiple...
In our chemical investigation into sp. UJNMF0740 derived from mangrove sediment, fourteen indole diterpene analogs, including four new ones, are purified by multiple chromatographic separation methods, with their structures being elucidated by the analyses of NMR, HR-ESIMS, and ECD data. The antibacterial and neuroprotective effects of these isolates were examined, and only compounds and exhibited weak antibacterial activity, while compounds , , and showed protective effects against the injury of PC12 cells induced by 6-hydroxydopamine (6-OHDA). Additionally, compound could suppress the apoptosis and production of reactive oxygen species (ROS) in 6-OHDA-stimulated PC12 cells as well as trigger the phosphorylation of PI3K and Akt. Taken together, our work enriches the structural diversity of indole diterpenes and hints that compounds of this skeleton can repress the 6-OHDA-induced apoptosis of PC12 cells via regulating the PI3K/Akt signaling pathway, which provides evidence for the future utilization of this fascinating class of molecules as potential neuroprotective agents.
Topics: Rats; Animals; PC12 Cells; Proto-Oncogene Proteins c-akt; Oxidopamine; Phosphatidylinositol 3-Kinases; Penicillium; Reactive Oxygen Species; Apoptosis; Diterpenes; Indoles; Anti-Bacterial Agents; Neuroprotective Agents
PubMed: 37999417
DOI: 10.3390/md21110593 -
Chemical & Pharmaceutical Bulletin Sep 2017Extracts of the sponge Hyattella aff. intestinalis showed moderate inhibition against adenovirus. Chromatographic separation of the extracts followed by analysis of...
Extracts of the sponge Hyattella aff. intestinalis showed moderate inhibition against adenovirus. Chromatographic separation of the extracts followed by analysis of spectroscopic data allowed us to elucidate the structures of three new metabolites as 2α-hydroxyspongia-13(16),14-diene-3-one (4), 3β-hydroxyspongia-13(16),14-diene-2-one (7), and 2α,3α-diacetoxy-17,19-dihydroxyspongia-13(16),14-diene (8) and to identify six known ones 1-3, 5, 6 and 9. Among the molecules, compounds 1 and 3 showed antiviral activity at IC 17.0 and 52.0 µM.
Topics: Adenoviridae; Animals; Antiviral Agents; Cell Line, Tumor; Cell Survival; Diterpenes; Inhibitory Concentration 50; Magnetic Resonance Spectroscopy; Molecular Conformation; Porifera; Rats
PubMed: 28652548
DOI: 10.1248/cpb.c17-00297