Structurally diverse (9ß-H)-pimarane derivatives with six frameworks from the leaves of Icacina oliviformis and their cytotoxic activities.

Thirteen previously undescribed (9ß-H)-pimarane derivatives, icacinolides A-G (1-7) and oliviformislactones C-H (8-13), together with four known analogs (14-17), were isolated from the leaves of Icacina oliviformis. Their structures were constructed by extensive spectroscopic analysis, 13C NMR-DP4+ analysis, ECD calculation, single-crystal X-ray diffraction, and chemical methods. These structurally diverse isolates were classified into six framework types: rearranged 3-epi-17-nor-(9ß-H)-pimarane, rearranged 17-nor-(9ß-H)-pimarane, 16-nor-(9ß-H)-pimarane, 17-nor-(9ß-H)-pimarane, 17,19-di-nor-(9ß-H)-pimarane, and (9ß-H)-pimarane. Among them, compounds 1, 5, and 7 were the first examples of three rearranged 3-epi-17-nor-(9ß-H)-pimaranes featuring a unique (11S)-carboxyl-9-oxatricyclo[5.3.1.02,7]dodecane motif with contiguous stereogenic centers, whereas their C-3 epimers, compounds 2-4 and 6 were the second examples of four rearranged 17-nor-(9ß-H)-pimaranes. Additionally, compounds 8 and 12/13 represented the second examples of a 16-nor-(9ß-H)-pimarane and two 17,19-di-nor-(9ß-H)-pimaranes, respectively. In cytotoxic bioassay, compound 2 exhibited significant cytotoxic against HT-29 with IC50 values of 7.88 µM, even stronger than 5-fluorouracil, and 15 showed broad-spectrum cytotoxic activities against HepG2, HT-29, and MIA PaCa-2 with IC50 values of 11.62, 9.77, and 4.91 µM, respectively. Meanwhile, a preliminary structure-activity relationship suggested that 3,20-epoxy, 6,19-lactone, 2-OH, 7-OH, and 8-OH in (9ß-H)-pimarane derivatives might be active groups, whereas ring C aromatization may decrease the cytotoxic activities.

New 21-nordammarane saponins with anti-inflammatory and cytotoxic activities from Lavigeria macrocarpa Oliv.

Chemical investigation of the ethanol extract from the stems and roots of the medicinal plant Lavigeria macrocarpa led to the isolation and structure elucidation of three previously unreported 21-nordammarane-type saponins namely 6α,27-dihydroxy-3,20-dioxo-21-nordammar-24-(Z)-ene 27-O-[α-L-rhamnopyranosyl-(1→2)-ß-D-glucopyranoside] (1), 6α,27-dihydroxy-3-oxo-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (2), and 2α,3ß,6α,27-tetrahydroxy-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (3) trivially named lavigemacrocarposide A-C, along with eight known secondary metabolites. Acid hydrolysis of lavigemacrocarposide A yielded a new prosapogenin namely 6α,27-dihydroxy-3,20-dioxo-21-nordammar-24-(Z)-ene 27-O-ß-D-glucopyranoside (1a) and the previously unreported artefactual aglycones 1b and 1c. Their structures were elucidated by spectroscopic analyses including mass spectrometry, 1D and 2D NMR as well as chemical evidence. The EtOH extract, some isolated compounds as well as the prosapogenin (1a) and compounds 1b and 1c were evaluated for anti-inflammatory and cytotoxic activity. Icacine (5) exhibited a significant cytotoxicity against both HeLa and MCF-7 cell lines with an IC50 value of 0.78 µg/mL. All the tested compounds showed more that 50% inhibition of NO production, except for 1 and 2.

Icacinaceae Plant Family: A Recapitulation of the Ethnobotanical, Phytochemical, Pharmacological, and Biotechnological Aspects.

Icacinaceae, an Angiospermic family comprising 35 genera and 212 accepted species, including trees, shrubs, and lianas with pantropical distribution, is one of the most outshining yet least explored plant families, which despite its vital role as a source of pharmaceuticals and nutraceuticals has received a meagre amount of attraction from the scientific community. Interestingly, Icacinaceae is considered a potential alternative resource for camptothecin and its derivatives, which are used in treating ovarian and metastatic colorectal cancer. However, the concept of this family has been revised many times, but further recognition is still needed. The prime objective of this review is to compile the available information on this family in order to popularize it in the scientific community and the general population and promote extensive exploration of these taxa. The phytochemical preparations or isolated compounds from the Icacinaceae family have been centrally amalgamated to draw diverse future prospects from this inclusive plant species. The ethnopharmacological activities and the associated endophytes and cell culture techniques are also depicted. Nevertheless, the methodical evaluation of the Icacinaceae family is the only means to preserve and corroborate the folkloristic remedial effects and provide scientific recognition of its potencies before they are lost under the blanket of modernization.

New diterpenoids and phenolic amides from Icacina mannii Oliv.

Fourteen undescribed compounds including five neoclerodanes (1-5), three labdanes (12-14), three pimarane (15-17) derivatives, one carbamate (24) and two clovamide-type amides (25 and 26), along with twenty-two known compounds (6-11, 18-23 and 27-36) were isolated from the tuber and stem of Icacina mannii. Their structures were elucidated by 1D and 2D NMR and HR-ESI-MS data analysis, and by comparing their NMR data with those of the literature.

Phenolic Glycosides from the Leaves of Iodes cirrhosa Turcz. with Cytotoxic and Antimicrobial Effects.

In the present study, 14 phenolic glycosides, including one new neolignan glycoside, iodescirrhoside A (1), three new flavonol glycosides, iodescirrhosides B-D (2-4), and 10 known metabolites were obtained from the methanol extract of Iodes cirrhosa leaves. Structural elucidation was performed by interpretating the 1D- and 2D- NMR, HR-ESI-MS, and CD spectra in comparison with literature data. All compounds were noncytotoxic to LU-1, HepG2, MCF-7, SK-Mel-2, and LNCaP cancer cell lines. Compound 11 significantly inhibited the growth of E. faecalis. In contrast, weak inhibition was observed for 1-9 and 14 against E. faecalis, for 1, 6-8, and 11 against S. aureus, for 6 and 13 against B. cereus, and 2, 4, and 6-9 against C. albicans.

19-nor-pimaranes from Icacina trichantha.

Four new unusual 19-nor-pimarane-type diterpenes were isolated from the tuber of Icacina trichantha (Icacinaceae, Oliv.). The structures were elucidated based on spectroscopic and HRMS analysis. The absolute configurations were determined by electronic circular dichroism. All four compounds are structural analogues of icacinol and humirianthol, but do not demonstrate the same cytotoxic activity. A plausible biogenetic pathway is proposed.

Systematics and phylogeny of Oecopetalum (Metteniusaceae), a genus of trees endemic to North and Central America

El género Oecopetalum Greenm. & C.H. Thomps. (Metteniusaceae) se distribuye en el sureste de México y en América Central (Guatemala, Nicaragua y Costa Rica). Tres especies han sido descritas. Estudiamos especímenes de herbario de Oecopetalum y realizamos un análisis filogenético basado en los genes de los cloroplastos matK y ndhF para responder a las preguntas principales: ¿Cuántas especies hay en el género Oecopetalum? ¿Es Oecopetalum monofilético? ¿Cuáles son las relaciones genéricas con otros miembros de la familia? Oecopetalum es un género monofilético con solo dos especies y Pittosporosis es el grupo hermano. La relación transatlántica de Oecopetalum y Pittosporosis es un patrón geográfico recurrente en la familia Mettenuisaceae, así como en Icacinaceae. Nuestros resultados, en concordancia con la evidencia fósil y las relaciones de otros grupos, apoyan el modelo boreotropical de migraciones terrestres de táxones tropicales desde latitudes altas durante el Paleoceno-Eoceno, globalmente cálido.

Oecopetalum Greenm. & C.H. Thomps. (Metteniusaceae) is distributed in the southeastern portion of Mexico to Central America (Guatemala, Nicaragua, and Costa Rica). Three species have been described. We studied herbarium specimens of the genus Oecopetalum and performed a phylogenetic analysis based on the plastid genes matK and ndhF to answer several major questions: How many species are in the genus Oecopetalum? Is Oecopetalum monophyletic, and how is the genus related to other members of the family? Our results indicate that Oecopetalum is monophyletic, with only two species, and sister to the Asian genus Pittosporopsis. The Trans-Atlantic relationship of Oecopetalum and Pittosporosis is a recurrent geographic pattern in the families Mettenuisaceae and Icacinaceae. Our results, in agreement the fossil record and previous phylogenetic studies, support the boreotropical model of high-latitude terrestrial migrations of tropical taxa during the globally warm Paleocene-Eocene.

Complete plastome sequence of Iodes cirrhosa Turcz., the first in the Icacinaceae, comparative genomic analyses and possible split of Idoes species in response to climate changes.

Plastome-based phylogenetic study has largely resolved the phylogeny of Icacinaceae. However, no single complete plastome sequence is available for Icacinaceae species, thereby limiting the further phylogenomics analysis of the members of this family. Here, we obtained the complete plastome sequence of Iodes cirrhosa Turcz., which is the first in Icacinaceae, by using the next-generation sequencing technology. The genome was annotated and compared with other closely related plastomes by using mVISTA. The divergence time of six Iodes species was analyzed using the BEAST software. The plastome of I. cirrhosa was 151,994 bp long, with a pair of inverted repeats (IRs, 24,973 bp) separated by a large single-copy (LSC, 84,527 bp) region and a small single-copy (SSC, 17,521 bp) region. The plastome encoded 112 unique genes, including 80 protein-coding, 28 tRNA, and four rRNA genes. Approximately 59 repeat sequences and 188 simple sequence repeats were identified. Four pairs of partially overlapped genes, namely, psbD/psbC, ndhF/Ψycf1, atpB/atpE, and rpl22/rps3, were observed. A comparison of the boundaries of the LSC, SSC, and IR regions with four other plastomes from Aquifoliales and Sapindales exhibited a high overall degree of sequence similarity. Four most highly variable regions, namely, trnH-GUG/psbA, psbM/trnD-GUC, petA/psbJ, and rps16/trnQ-UUG, were found. Using the plastome of I. cirrhosa as reference, we reassembled the plastomes of five Iodes species. K a/K s ratio analyses revealed that 27 genes and 52 amino acid residue sites from 11 genes had undergone strong positive selection in the Iodes branch, with the most abundant proteins being the NDH and ribosomal proteins. Divergence-time analysis indicated that Iodes species were first formed 34.40 million years ago. Results revealed that the ancestor of the six species was likely to have split in the late Eocene epoch. In summary, the first complete plastome sequence of I. cirrhosa provided valuable information regarding the evolutionary processes of Iodes species.

Mappianines A-E, structurally diverse monoterpenoid indole alkaloids from Mappianthus iodoides.

Five previously undescribed monoterpenoid indole alkaloids, mappianines A-E, along with twelve known analogues, were isolated from the stems of Mappianthus iodoides Hand.-Mazz. Their structures and absolute configurations were determined by spectroscopic analysis, single-crystal X-ray diffraction, and ECD calculations. The plausible biogenetic pathway of mappianine A was proposed. All the isolated compounds were evaluated for their cytotoxic effects on MGC-803, Bel-7404, A549, NCI-H460, and HepG2 cancer cell lines. Mappianine B, tetrahydroalstonine, ß-carbolin-1-one, and 1,2,3,4-tetrahydronorharman-1-one displayed moderate cytotoxicity against all cell lines tested, with IC50 values ranging from 5.19 to 42.86 µM.

Chemical constituents from stems and leaves of Mappianthus iodoides / 中草药

Objective To study the chemical constituents of the stems and leaves of Mappianthus iodoides. Methods The chemical constituents of M. iodoides were separated and purified by silica gel, ODS, Sephadex LH-20 gel column chromatographies, and preparative HPLC. Their structures were identified by physicochemical properties, spectroscopic analysis, as well as comparisons with the data reported in literature. Results Sixteen compounds were isolated from the 90% ethanol extract of the stems and leaves of M. iodoides, which were identified as 9-hydroxy-4,6-megastigmadien-3-one (1), 9-hydroxy-4,7-megastigmadien-3-one (2), blumenol A (3), 9,10-dihydroxy-4,7-megastigmadien-3-one (4), 5,12-epoxy-9-hydroxy-7-megastigmen-3-one (5), 5,12-epoxy-6,9-hydroxy-7- megastigmen-3-one (6), loliolide (7), lariciresinol (8), isolariciresinol (9), 5′-methoxylariciresinol (10), olivil (11), dehydrodiconiferyl alcohol (12), chushizisin I (13), 3,3-didemethoxyverrucosin (14), 4-epi-larreatricin (15), and glycerol monolinoleate (16). Conclusion All compounds are isolated from the genus Mappianthus for the first time.

Biological activities of triterpenoids from Poraqueiba sericea stems.

Eleven compounds were isolated from Poraqueiba sericea stems and identified as niga-ichigoside-F1 (1), trachelosperoside B1 (2), 4-epi-niga-ichigoside (7), 19α-hydroxyasiatic acid (3), myrianthic acid (4), hyptatic acid (5), trachelosperogenin B (6), arjunolic acid (8), and trachelosperogenin E (9), secologanoside (10) and secoxyloganin (11). Compounds 1-11 were tested for their antileishmanial activities against Leishmania infantum promastigotes, 1-6 and 8-11 were tested for their cytotoxic activities on fibroblasts, 1-3, 5-6, 8-11 were evaluated for their anti-elastase and anti-acetylcholinesterase assays activities by a spectrophotometric method and 1-2, 5 and 7-10 were tested using bioautography for their ß-glucosidase. No antileishmanial activity was detected; compounds 1, 2 and 11 showed a moderate cytotoxic activity with IC50 17.7, 20.5 and 10.9 µg/mL, respectively; compounds 2, 8, 9 and 10 gave a percentage of inhibition ranging from 13 to 16% (at 50 µg/mL) and compounds 1 and 2 showed an inhibition zone on ß-glucosidase and anti-acetylcholinesterase assays.

Resolving basal lamiid phylogeny and the circumscription of Icacinaceae with a plastome-scale data set.

PREMISE OF THE STUDY: Major relationships within Lamiidae, an asterid clade with ∼40000 species, have largely eluded resolution despite two decades of intensive study. The phylogenetic positions of Icacinaceae and other early-diverging lamiid clades (Garryales, Metteniusaceae, and Oncothecaceae) have been particularly problematic, hindering classification and impeding our understanding of early lamiid (and euasterid) character evolution. METHODS: To resolve basal lamiid phylogeny, we sequenced 50 plastid genomes using the Illumina sequencing platform and combined these with available asterid plastome sequence data for more comprehensive phylogenetic analyses. KEY RESULTS: Our analyses resolved basal lamiid relationships with strong support, including the circumscription and phylogenetic position of the enigmatic Icacinaceae. This greatly improved basal lamiid phylogeny offers insight into character evolution and facilitates an updated classification for this clade, which we present here, including phylogenetic definitions for 10 new or converted clade names. We also offer recommendations for applying this classification to the Angiosperm Phylogeny Group (APG) system, including the recognition of a reduced Icacinaceae, an expanded Metteniusaceae, and two orders new to APG: Icacinales (Icacinaceae + Oncothecaceae) and Metteniusales (Metteniusaceae). CONCLUSIONS: The lamiids possibly radiated from an ancestry of tropical trees with inconspicuous flowers and large, drupaceous fruits, given that these morphological characters are distributed across a grade of lineages (Icacinaceae, Oncothecaceae, Metteniusaceae) subtending the core lamiid clade (Boraginales, Gentianales, Lamiales, Solanales, Vahlia). Furthermore, the presence of similar morphological features among members of Aquifoliales suggests these characters might be ancestral for the Gentianidae (euasterids) as a whole.

Icacinaceae from the eocene of Western North America.

PREMISE OF THE STUDY: The Icacinaceae are a pantropical family of trees, shrubs, and climbers with an extensive Paleogene fossil record. Our improved understanding of phylogenetic relationships within the family provides an excellent context for investigating new fossil fruit and leaf material from the Eocene of western North America. METHODS: We examined fossils from early and middle Eocene sediments of western Wyoming, northeastern Utah, northwestern Colorado, and Oregon and compared them with extant species of Iodes and other icacinaceous genera as well as previously described fossils of the family. KEY RESULTS: Three new fossil species are described, including two based on endocarps (Iodes occidentalis sp. nov. and Icacinicaryites lottii sp. nov.) and one based on leaves (Goweria bluerimensis sp. nov.). The co-occurrence of I. occidentalis and G. bluerimensis suggests these might represent detached organs of a single species. A new genus, Biceratocarpum, is also established for morphologically distinct fossil fruits of Icacinaceae previously placed in Carpolithus. Biceratocarpum brownii gen. et comb. nov. resembles the London Clay species "Iodes" corniculata in possessing a pair of subapical protrusions. CONCLUSIONS: These fossils increase our knowledge of Icacinaceae in the Paleogene of North America and highlight the importance of the Northern Hemisphere in the early diversification of the family. They also document interchange with the Eocene flora of Europe and biogeographic connections with modern floras of Africa and Asia, where Icacinaceae are diverse today. The present-day restriction of this family to tropical regions offers ecological implications for the Eocene floras in which they occur.

Structural insight of DNA topoisomerases I from camptothecin-producing plants revealed by molecular dynamics simulations.

DNA topoisomerase I (Top1) catalyzes changes in DNA topology by cleaving and rejoining one strand of the double stranded (ds)DNA. Eukaryotic Top1s are the cellular target of the plant-derived anticancer indole alkaloid camptothecin (CPT), which reversibly stabilizes the Top1-dsDNA complex. However, CPT-producing plants, including Camptotheca acuminata, Ophiorrhiza pumila and Ophiorrhiza liukiuensis, are highly resistant to CPT because they possess point-mutated Top1. Here, the adaptive convergent evolution is reported between CPT production ability and mutations in their Top1, as a universal resistance mechanism found in all tested CPT-producing plants. This includes Nothapodytes nimmoniana, one of the major sources of CPT. To obtain a structural insight of the resistance mechanism, molecular dynamics simulations of CPT- resistant and -sensitive plant Top1s complexed with dsDNA and topotecan (a CPT derivative) were performed, these being compared to that for the CPT-sensitive human Top1. As a result, two mutations, Val617Gly and Asp710Gly, were identified in O. pumila Top1 and C. acuminata Top1, respectively. The substitutions at these two positions, surprisingly, are the same as those found in a CPT derivative-resistant human colon adenocarcinoma cell line. The results also demonstrated an increased linker flexibility of the CPT-resistant Top1, providing an additional explanation for the resistance mechanism found in CPT-producing plants. These mutations could reflect the long evolutionary adaptation of CPT-producing plant Top1s to confer a higher degree of resistance.

Floral structure of Emmotum (Icacinaceae sensu stricto or Emmotaceae), a phylogenetically isolated genus of lamiids with a unique pseudotrimerous gynoecium, bitegmic ovules and monosporangiate thecae.

BACKGROUND AND AIMS: Icacinaceae sensu stricto consist of a group of early branching lineages of lamiids whose relationships are not yet resolved and whose detailed floral morphology is poorly known. The most bizarre flowers occur in Emmotum: the gynoecium has three locules on one side and none on the other. It has been interpreted as consisting of three fertile and two sterile carpels or of one fertile carpel with two longitudinal septa and two sterile carpels. This study focused primarily on the outer and inner morphology of the gynoecium to resolve its disputed structure, and ovule structure was also studied. In addition, the perianth and androecium were investigated. METHODS: Flowers and floral buds of two Emmotum species, E. harleyi and E. nitens, were collected and fixed in the field, and then studied by scanning electron microscopy. Microtome section series were used to reconstruct their morphology. KEY RESULTS: The gynoecium in Emmotum was confirmed as pentamerous, consisting of three fertile and two sterile carpels. Each of the three locules behaves as the single locule in other Icacinaceae, with the placenta of the two ovules being identical, which shows that three fertile carpels are present. In addition, it was found that the ovules are bitegmic, which is almost unique in lamiids, and that the stamens have monosporangiate thecae, which also occurs in the closely related family Oncothecaceae, but is not known from any other Icacinaceae sensu lato so far. CONCLUSIONS: The flowers of Emmotum have unique characters at different evolutionary levels: the pseudotrimerous gynoecium at angiosperm level, the bitegmic ovules at lamiid level and the monosporangiate thecae at family or family group level. However, in general, the floral morphology of Emmotum fits well in Icacinaceae. More comparative research on flower structure is necessary in Icacinaceae and other early branching lineages of lamiids to better understand the initial evolution of this large lineage of asterids.

Pyrenacantha volubilis Wight, (Icacinaceae) a rich source of camptothecine and its derivatives, from the Coromandel Coast forests of India.

Camptothecine, a potent eukaryotic topoisomerase inhibitor, is an important anticancer compound. The global demand for this compound was estimated to be $1 billion in 2003 and is only further expected to increase. Partly to meet the expected increase in demand, in the recent past, several efforts have been made to discover newer and alternative plant and fungal sources of camptothecine. In this study we report a rich source of camptothecine and its natural derivatives, Pyrenacantha volubilis (Icacinaceae) from the eastern coast of peninsular India. Camptothecine and its derivatives were analyzed using high performance liquid chromatography (HPLC) coupled with electrospray mass spectrometry (ESI-MS) in all plant parts such as twigs, leaves, roots, seedling, ripened whole fruit, fruit coat, seed coat and cotyledons. Cotyledons and ripened whole fruits contained the highest amount of camptothecine (1.35% and 0.60% dry weight respectively). LC-MS and ESI-MS/MS analyses revealed besides camptothecine, other derivatives and precursors such as 10-hydroxycamptothecine, 9-methoxycamptothecine, 20-deoxycamptothecine, deoxypumiloside, strictosidine and strictosamide. Pure camptothecine was isolated from fruits and structurally confirmed using NMR. Seed extracts were found to be effective against breast cancer, ovarian, colon and carcinoma cell lines (with IC50 values of 4.0 µg/mL, 6.5 µg/mL, 25.0 µg/mL and 25.0 µg/mL respectively). We discuss the results in the context of exploring alternative sources of camptothecine.

Terpenoid indole alkaloids from Mappianthus iodoides Hand.-Mazz.

Ten terpenoid indole alkaloids, mappiodines A-C and mappiodosides A-G, together with eight known compounds, were isolated from stems of Mappianthus iodoides Hand.-Mazz. Their structures were elucidated by spectroscopic analyses including 1D, 2D NMR, MS and CD methods. The ten compounds were evaluated for their cytotoxic activity, but were inactive.

HPLC analysis of camptothecin content in various parts of Nothapodytes foetida collected on different periods.

OBJECTIVE: To investigate the content of topoisomerase I-DNA inhibitor alkaloid camptothecin (CPT) from various parts of Nothapodytes foetida (N. foetida) collected from the month of October to February. METHODS: The content of CPT was quantified in the methanolic extract of various parts of N. foetida using high performance liquid chromatography (HPLC). Quantification was performed with the regression analysis and the method was validated as per ICH guidelines. RESULTS: The results revealed that maximum concentrations of camptothecin were found in root (2.62%) collected in the month of February followed by fruits (January, 1.22%), stem (January, 0.81%) and leaves (February, 0.70%). Roots were found to have 3-fold higher concentration of CPT than the leaves and stem, while the fruits showed 2-fold higher concentration. Maximum concentration of camptothecin in fruits was observed in month of January, when they were not fully mature, which was 2-fold higher than that of young and fully mature fruits. CONCLUSIONS: These findings indicate that the synthesis of CPT differs in different parts of N. foetida and the content varies periodically.