Ingenane diterpenoids from Euphorbia esula

An extensive study of metabolites present in Euphorbia esula led to isolation of 16 ingenane diterpenoids 1-16 together with the known ingenane derivative 17 and four known cycloartane triterpenoids. Their structures were elucidated on the basis of spectroscopic studies and comparison with known related compounds. All the compounds were assayed for their inhibitory activity against human HeLa cervical cancer cell line.     

Production of Bioactive Diterpenoids in the Euphorbiaceae Depends on Evolutionarily Conserved Gene Clusters

The Euphorbiaceae produce a diverse range of diterpenoids, many of which have pharmacological activities. These diterpenoids include ingenol mebutate, which is licensed for the treatment of a precancerous skin condition (actinic keratosis), and phorbol derivatives such as resiniferatoxin and prostratin, which are undergoing investigation for the treatment of severe pain and HIV, respectively. Despite the interest in these diterpenoids, their biosynthesis is poorly understood at present, with the only characterized step being the conversion of geranylgeranyl pyrophosphate into casbene. Here, we report a physical cluster of diterpenoid biosynthetic genes from castor (Ricinus communis), including casbene synthases and cytochrome P450s from the CYP726A subfamily. CYP726A14, CYP726A17, and CYP726A18 were able to catalyze 5-oxidation of casbene, a conserved oxidation step in the biosynthesis of this family of medicinally important diterpenoids. CYP726A16 catalyzed 7,8-epoxidation of 5-keto-casbene and CYP726A15 catalyzed 5-oxidation of neocembrene. Evidence of similar gene clustering was also found in two other Euphorbiaceae, including Euphorbia peplus, the source organism of ingenol mebutate. These results demonstrate conservation of gene clusters at the higher taxonomic level of the plant family and that this phenomenon could prove useful in further elucidating diterpenoid biosynthetic pathways.     

Oil body formation in Euphorbia tirucalli L. cell suspension cultures

The culture of cells of Euphorbia tirucalli L. resulted in the formation of intracellular particles which osmium tetroxide staining and electron microscopy indicated to be oil bodies. Chemical analysis (TLC, HPLC, GLC and GC-MS) disclosed steroids, triterpenoids, and diterpenoids as the major oil components of the cultured cells.     

A comparative phytochemical study of the diterpenes of some species of the genera Euphorbia and Elaeophorbia (Euphorhiaceae)

Nearly 60 species from the genera Euphorbia L. and Elaeophorbia Stapf were investigated for diterpenes of the classes tiglianes, ingenanes and ortho-esters. The diterpenes were isolated as their acetates by a micro-technique from latex or fresh herb material collected from several countries around the world. Authentication of the diterpenes was by chromatographic means using spectroscopic methods as back-up data. These compounds were absent from only 9% of the species examined. The most commonly occurring diterpene was ingenol, followed closely by 12-Deoxy phorbol. The ingenane derivative 5-Deoxy-ingenol was always detected as a minor companion to ingenol from species of the section Tithymalus. Species of the section Euphorbium contained mainly the diterpene ingenol although both tigliane and ortho-ester diterpenes were also present in some species. Species from the sections Anisophylhtm and Poinsettia did not contain diterpenes of the type in question, whilst species from the genus Elaeophorbia yielded ingenol. These results provide additional chemical evidence concerning recent suggestions on the subgeneric and generic status of Anisophyllum, Poinsettia, Tithymalus and Elaeophorbia.     

Quantitative Analysis of Ingenol in Euphorbia species via Validated Isotope Dilution Ultra‐high Performance Liquid Chromatography Tandem Mass Spectrometry

Introduction

Various species of the Euphorbia genus contain diterpene ingenol and ingenol mebutate (ingenol‐3‐angelate), a substance found in the sap of the plant Euphorbia peplus and an inducer of cell death. A gel formulation of the drug has been approved by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for the topical treatment of actinic keratosis.

Objective

To develop a rapid and reliable method for quantification of ingenol in various plant extracts.

Methodology

Methanolic extracts of 38 species of the Euphorbia genus were analysed via ultra‐high performance liquid chromatography with tandem mass spectrometry (UHPLC–MS/MS) after methanolysis and solid‐phase extraction (SPE) purification. The ¹⁸O–labelled ingenol analogue was prepared and used as an internal standard for ingenol content determination and method validation.

Results

The highest ingenol concentration (547 mg/kg of dry weight) was found in the lower leafless stems of E. myrsinites. The screening confirms a substantial amount of ingenol in species studied previously and furthermore, reveals some new promising candidates.

Conclusion

The newly established UHPLC–MS/MS method shows to be an appropriate tool for screening of the Euphorbia genus for ingenol content and allows selection of species suitable for raw material production and/or in vitro culture initiation. Copyright © 2017 John Wiley & Sons, Ltd.     

Diterpenoids and triterpenoids from Euphorbia guyoniana

Two new compounds with tigliane and cycloartane skeletons: 4,12-dideoxy(4alpha)phorbol-13-hexadecanoate (1) and 24-methylenecycloartane-3,28-diol (2), respectively, in addition of four known diterpenoids and 13 triterpenoids: 3-benzoyloxy-5,15-diacetoxy-9,14-dioxojatropha-6(17),11-diene (4), ent-abieta-8(14),13(15)-dien-16,12-olide (5), ent-8alpha,14alpha-epoxyabieta-11,13(15)-dien-16,12-olide (6), ent-3-hydroxyatis-16(17)-ene-2,14-dione (7), 3beta-hydroxytaraxer-14-en-28-oic acid (8), beta-sitosteryl-3beta-glucopyranoside-6'-O-palmitate (9), multiflorenyl acetate (10), multiflorenyl palmitate (11), peplusol (12), 24-methylenecycloartanol (3), lanosterol (13), euferol (14), butyrospermol (15), cycloartenol (16), obtusifoliol (17), cycloeucalenol (18) and beta-sitosterol (19), were isolated from the roots of Euphorbia guyoniana. Their structures were established on the basis of physical and spectroscopic analysis, including 1D and 2D homo- and heteronuclear NMR experiments (COSY, HSQC, HMBC and NOESY) and by comparison with the literature data.     

Identification and characterisation of the chinese herb Langdu by LC-MS/MS analysis

An LC-MS/MS method has been developed for the identification of three species of herb used as the traditional Chinese medicine Langdu, namely Stellera chamaejasme L., Euphorbia ebracteolata Hayata and E. fischeriana Steud. As these herbs contain different mixtures of marker compounds, they could be unambiguously differentiated from each other by comparing their respective characteristic segmental multiple reaction monitoring profiles. The profiles indicated that S. chamaejasme contained daphnetin, skimmetine, stellerin, chamaechromone and neochamaejasmin, E. fischeriana contained ebracteolata compound B, ingenol, jolkinolide B and fischeriana A, whilst E. ebracteolata contained ebracteolata compounds B and C along with ingenol. These results were confirmed from the respective MS/MS spectra. The method has been successfully applied to differentiate these herbs from the related species Alocasia macrorrhiza (L.) Schott and E. kansui Liou.     

Tumor promoting diterpenes from Euphorbia leuconeura L.

Diterpene esters of the phorbol and ingenol types are known to be highly active tumor promoting agents that typically occur in members of the Euphorbiaceae. In the present work, Euphorbia leuconeura, a rare indoor plant, is analyzed for its tumor promoting potential. Latex as well as total leaf extracts exhibited Epstein-Barr-virus (EBV) inducing activity comparable to 12-O-tetradecanoyl-phorbol-13-O-acetate, a well known tumor promoter. The activity of individual fractions correlated with their ingenol ester content. Three ingenol esters with EBV inducing activity could be isolated and identified. They belong to the milliamine type of diterpene esters that contain aromatic peptidyl groups. Two of them (milliamines L and M) are already known from E. milii. The third compound is identified as an isomer of milliamine F with a novel 3,20-diester arrangement. The data show a close relationship between E. leuconeura and the more popular indoor plant E. milii whose latex is also used as a powerful molluscicide.     

Bioactive diterpenoids from Celastraceae species

Celastraceae species have a long tradition of use in folk medicine and agriculture. Intensive research into this family has resulted in the isolation of a large number of secondary metabolites with a wide range of bioactivity, with the most characteristic being the dihydro-β-agarofuran sesquiterpenes and the quinomethide triterpenoids. Despite diterpenoids not often being found in Celastraceae species, the novelty of their structures and their biological activities has stimulated their research. There are several reviews covering studies on Tripterigium wilfordii, a traditional Chinese medicine, and their metabolites, but none focusing on bioactive diterpenoids isolated from other Celastraceae species. This review focuses on diterpenes isolated from Celastraceae species, and their skeleton diversity, species sources, and tested biological activities are discussed. The literature from January 2000 to October 2016 is reviewed, and 64 references are cited. A total of 118 isolated diterpenoids possessing twelve different skeletons are included and classified by the following tested biological activity: cytotoxicity, anti-inflammatory, antitumour-promotion, anti-HIV, immunosuppressant, reversal multi-drug resistance, and antifeedant. This compliance information will be helpful in further research into diterpene Celastraceae isolation, biological activity determination and structure specific modifications.     

海绵的生理活性代谢产物—萜类的研究进展

海绵中含有多种多样的萜类化合物,包括单萜、倍半萜、二萜、二倍半萜和三萜。其中,倍半萜和二萜最为丰富。这些次生代谢物大部分具有强烈的生理活性。本文介绍近年来这一领域的研究进展。     

Cytotoxic clerodane diterpenoids from the leaves of Premna tomentosa

Three clerodane diterpenoids, premnones A-C (1-3), were isolated from a chloroform-soluble fraction of Premna tomentosa along with four known flavonoids and three known triterpenoids. Among these isolates, premnones A-C exhibited cytotoxic activity when evaluated against a small panel of tumor cell lines. However, premnone A was found to be inactive when evaluated in a follow-up in vivo hollow fiber assay at the highest dose tested (50mg/kg), using LNCaP, Lu1, and MCF-7 cells.     

Chrysobalanaceae: secondary metabolites, ethnopharmacology and pharmacological potential

Many Chrysobalanaceae species, in special Licania and Parinari, are widely used in folk medicine to treat several diseases. This review describes some aspects of their ethnopharmacology potential, biological activities and the secondary metabolites reported so far for Chrysobalanaceae. The chemical constituents of this family include triterpenoids, diterpenoids, steroids and phenylpropanoids like flavonoids as well as chromones derivatives.     

Diterpenoids from Euphorbia paralias.

Two segatane diterpenoids containing a bicyclic [4,3,1] ring system were isolated, together with 11 known diterpenoids, four segetanes, five jatrophanes and two paralianes, from the aerial parts of Euphorbia paralias. The structures of the new compounds were established by spectroscopic means, including by 2D NMR and CD spectroscopic analyses. The antifeedant and antiviral activities of the isolated compounds are also described.     

Four New Diterpenoids from the Roots of Euphorbia pekinensis

Three new isopimarane diterpenoids ( 1 – 3 ) and one new abietane diterpenoid ( 4 ) were isolated and identified from the roots of Euphorbia pekinensis, together with four known diterpenoids. Their structures were elucidated on the basis of extensive spectroscopic methods (IR, MS, NMR, and CD), and their cytotoxicities and anticomplement activities were evaluated.     

α-Onocerin and sterol content of twelve species of Ononis

The sterols and triterpenoids of 12 species of the genus Ononis were analysed by GLC. α-Onocerin was found in all but one of these species, although in some others its concentration was low. In all species examined, sitosterol was the major sterol; stigmasterol, campesterol, cholesterol and the triterpenoids cycloartenol and 24-methylene cycloartanol also occurred. The patterns of α-onocerin and sterols found seem to be consistent with the accepted classification of species within the genus.     

A new spiny, cushion-like Euphorbia (Euphorbiaceae) from south-west Iran with special reference to the phytogeographic importance of local endemic species

Euphorbia acanthodes Akhani is described as a new gypsophilous species from south-west Iran. The affinity, morphological and carpological features, and habitat of the species are discussed. A list of a further 36 interesting endemic species of south-western Iran and adjacent areas is given with notes on their affinities and distribution. These include Centaurea gudrunensis Boiss. & Hausskn., as a new record from Iran. The specific status of Pteropyrum naufelum Al-Khayat is confirmed and it is suggested that P. noëanum Boiss. ex Meisn. is synonymous with P. aucheri Boiss. The biogeographic importance of the so-called 'Persian foothills', and biodiversity conservation status of the area are discussed, together with distribution maps of 12 species. It is concluded that the majority of the endemic species in the southern and south-western parts of Iran are Irano–Turanian or have their origins in the Irano–Turanian region. Therefore, it is questionable to consider the area as part of either the Saharo–Sindian, Sudanian or Sudano–Zambezian regions.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 107–121.     

Ingenol esters from the pro-inflammatory fraction of Euphorbia kamerunica

A series of unstable mono- and di-esters of the tetracyclic diterpene ingenol were isolated from the pro-inflammatory ether-soluble fraction of the latex of Euphorbia kamerunica. The esters were isolated by a neutral process involving column and thin-layer chromatography. The monoesters were identified by spectroscopic methods and hydrolysis reactions as ingenol-3-decanoate, ingenol-3-dodecanoate, ingenol-5-hexadienoate and ingenol-5-octenoate and the diesters as 20-acetyl-ingenol-3-octenoate and 20-acetyl-ingenol-3-angelate.     

Triterpenoids from Euphorbia fischeriana

Nineteen triterpenoids including two new tirucallane ones, fisuphanins A and B, were isolated from the whole plant of Euphorbia fischeriana Steud. The new structures were elucidated on the basis of extensive spectroscopy techniques including infrared (IR), High Resolution Mass Spectrometry (HRMS), and Nuclear Magnetic Resonance (NMR). Some triterpenoids were evaluated for their cytotoxicity against A549 and MCF-7 cell lines.     

The genus Plectranthus in India and its chemistry

Phytochemical constituents isolated from Indian species of the genus Plectranthus reported up to 2009 are compiled. In India, the genus Plectranthus is found in all the habitats and altitudes, particularly in the Himalaya, the Southern Ghats, and the Nilgiri region. P. amboinicus, P. barbatus, P. caninus, P. mollis, P. coetsa, and P. incanus are the most common species found in India. Phytochemical studies of the genus revealed that Indian Plectranthus species are rich in essential oil, and that the most abundant secondary metabolites are diterpenoids, i.e., labdanes, abietanes, and ent-kauranes, as well as triterpenoids.