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Stimulated by the promising results seen for its phenanthridones in clinical trials, interest in the Amaryllidaceae as a potential source of anticancer drugs has intensified in recent years. As such, over three hundred isoquinoline principles from its various alkaloid groups have to date been screened against a large number of cancer cell lines. Whilst significant effort has been diverted towards studies of lycorine and its congeners, the closely-allied homolycorine alkaloids of this plant family have largely been overlooked. These compounds have often been used as supporting acts to bolster the diversity of structures in cytotoxicity studies of the more recognized phenanthridone, lycorane and crinane alkaloids of the Amaryllidaceae. This notwithstanding, a substantial amount of information has surfaced over the past several years on such effects for the homolycorine alkaloids. This review takes a detailed look at the cytotoxic effects manifested by over forty of these alkaloids against around sixty cell lines, which may be classified into eighteen different types of cancers. In this regard, good activities were detected for some constituents (such as lycorenine and hippeastrine) against various hepatic and prostate cancers. The structural features which support and fortify the efficacy of the homolycorine alkaloid cytotoxic pharmacophore are also considered. These include the ways in which the innate ring systems are appended as well as the size, geometry and electronics of the attendant substituents. In addition, the various mechanisms used to rationalize the cytotoxic responses are described notably, interaction with DNA, apoptosis induction, protection from tumor invasion, efflux pump perturbation and topoisomerase inhibition.

From camptothecin to vinblastine, plant alkaloids continue to capture the imagination of scientists as a valuable source of anticancer drugs. Given that they are also amongst the most common secondary metabolites encountered in plants, the number of alkaloid entities subjected to cytotoxicity evaluations now runs into several thousands. Interest in the plant family Amaryllidaceae has been persistent owing to the potent cytotoxic activities of several of its isoquinoline alkaloid principles. This survey is an indepth account of such properties discernible for tazettine alkaloids belonging to the minor alkaloid groups of the Amaryllidaceae. Eleven of such compounds have up to now been screened against around 50 cancer cell lines which may be classified into nearly 20 different types of cancers. Although the activities in most instances were moderate to mild, notable responses were observed against some leukemia, adenocarcinoma, lymphoma and glioblastoma cell lines. In fact, pretazettine (ED50 0.3 μg/mL) was amongst the most potent of all Amaryllidaceae alkaloids screened against the human Molt4 T-lymphoma cell line. To most accounts, the parent compounds (such as tazettine and pretazettine) with their distinct ring systems and substitution patterns exhibit the best activities, offering little space for structural adjustments via semi-synthetic operations. Several avenues have been probed in attempts to elucidate the mechanisms by which tazettine alkaloids manifest their cytotoxic effects including, inhibition of protein synthesis, apoptosis and efflux pump interactions, the first of which to date offers the most profound insights.

Protozoan-borne diseases are prominent amongst diseases caused by parasites. Given their alarming morbidity and mortality statistics, there is ever growing interest in new therapies against these diseases. Whilst synthetic drugs such as benznidazole and melarsoprol have had a profound influence on the clinical setup, there has been significant interest in the phytochemical platform to also deliver such drug candidates. The plant family Amaryllidaceae is recognizable for its isoquinoline alkaloids, which exhibit attractive molecular architectures and interesting biological properties. This survey focuses on the antiprotozoal activities of 73 of such substances described in 18 different species of the Amaryllidaceae. Of these, 2-O-acetyllycorine was identified as the most potent (IC50 0.15 μg/mL against Trypansoma brucei brucei). Also considered are structure-activity relationships which have served to modulate activities, as well as the plausible mechanisms that underpin these effects and afford insight to the Amaryllidaceae alkaloid antiprotozoal pharmacophore.

The spread of malaria is thought to have followed human expansion out of Africa some 60 – 80 thousand years ago. With its prevalence in pantropical countries of the world and epicenter localized in Africa, malaria is now considered an unnecessary burden to overworked and under-resourced healthcare structures. Plants have long afforded a fertile hunting ground for the search and identification of structurally diverse antimalarial agents, such as quinine and artemisinin. This survey examines the antiparasitic properties of the family Amaryllidaceae via the antiplasmodial activities demonstrated for its lycorane alkaloid principles. Of these, 24 were natural compounds identified in 20 species from 11 genera of the Amaryllidaceae family, whilst the remaining 28 were synthetically derived entities based on the lycorane skeleton. These were screened against ten different strains of the malarial parasite Plasmodium falciparum, wherein the parent compound lycorine was shown to be the most potent with an IC50 of 0.029 µg/mL in the FCR-3 strain seen to be the best. Structure-activity relationship studies revealed that good activities were detectable across both the natural compounds as well as the synthetically accessed derivatives. Such studies also highlighted that there are several inherent structural features that define the lycorane alkaloid antiplasmodial pharmacophore, such as the nature of its ring systems and properties of its substituents. Mechanistically, a limited number of studies confirmed that lycorane alkaloids manifest their action by targeting enzymes associated with the plasmodial FAS-II biosynthetic pathways. Overall, these alkaloids have provided useful, convenient, and accessible scaffolds for antimalarial-based drug discovery.

A new phytochemical study of the indigenous Brazilian species Hippeastrum papilio is reported herein. Three novel Amaryllidaceae alkaloids were isolated, including hippapiline (1), papiline (2), and 3-O-demethyl-3-O-(3-hydroxybutanoyl)haemanthamine (3). Their structures were determined by physical and spectroscopic methods. In addition, the known alkaloids, haemanthamine (4), galanthamine (5), narwedine (6), 11β-hydroxygalanthamine (7), apogalanthamine (8), and 9-O-demethyllycosinine B (9) were identified. The unusual cis-B/C-ring fusion for the new homolycorine representative hippapiline was ratified by NMR and CD spectroscopy. Fil: Guo, Ying. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España Fil: Andrade, Jean P. de. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España. Universidade Federal do Espírito Santo; Brasil Fil: Pigni, Natalia Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España Fil: Torras-Claveria, Laura. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España. Universidade Federal do Espírito Santo; Brasil Fil: Tallini, Luciana R.. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España Fil: Souza Borges, Warley de. Universidade Federal do Espírito Santo; Brasil Fil: Viladomat, Francesc. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España Fil: Nair, Jerald J.. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España Fil: Zuanazzi, José A. S.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Bastida, Jaume. University of Barcelona. Faculty of Pharmacy. Department of Natural Products, Plant Biology and Soil Science; España

Representatives of the genus Cyrtanthus including Cyrtanthus contractus of the Amaryllidaceae family are widely used in African traditional medicine for the treatment of a range of ailments, including mental illness and age-related dementia. The alkaloid constituents distributed within the genus, which are chiefly responsible for the biological activity of the derived extracts, are thought to be influenced by changes in the growing environment. Natural growing populations of C. contractus were tagged and collected every 15th day of each month over a 12 month period. The ethanol bulb extracts from each of the collected samples were evaluated for total alkaloid content, cytotoxic effects, acetylcholinesterase and cyclooxygenase inhibition as well as antimicrobial activities. Some notable bioactivities were recorded for extracts collected during certain months of the year, particularly the cytotoxic effects against human cancer cell lines and enzyme inhibition (AChE and COX). The cytotoxic effects and AChE and COX enzyme inhibitory activities of extracts collected in May and September are of considerable interest. Similarly total alkaloids varied markedly from one month to the other. The bioactivity exhibited by C. contractus bulb extracts during certain months of the year, particularly in May and September, justifies the collection and use of these bulbs for treatment of the relevant ailments in traditional medicine during these periods.

The discovery of the karrikin class of plant growth regulators (PGRs) is a milestone accomplishment in plant biochemistry and physiology, with significant potential in agriculture and horticulture. These compounds have in common a fused furano-pyran ring system featuring various permutations of methyl substitution. Chief amongst these compounds is karrikinolide (KAR1), identified as the key germination stimulant present in plant-derived smoke, which together with five other closely-related structures (KAR2–KAR6) make up the karrikin class of PGRs. By contrast, the germination inhibitor 3,4,5-trimethyl-2(5H)-furanone has also been identified in plant-derived smoke. Various synthetic endeavours have been undertaken for structure–activity relationship study purposes as well as to probe the molecular mechanics of these compounds. In this study, syntheses of KAR1, KAR3 and S-KAR1 were carried out and their structures verified by X-ray crystallography. Effects on germination were measured against the inhibitor 3,4,5-trimethyl-2(5H)-furanone in Grand Rapids lettuce seeds. X-ray crystallographic data and germination promotory activity for S-KAR1 are described for the first time.

Apoptosis-inducing anticancer drugs have garnered widespread interest in recent years. Targets which concomitantly also exhibit minimal adverse effects in normal, healthy cells have been particularly well-received. The phenanthridone alkaloids such as pancratistatin and narciclasine exemplify such a class of chemotherapeutics, with potent and selective cytotoxic effects in a wide variety of cancer cells which are manifested via the apoptotic mode of cell death. Caspases are central to the apoptotic process through their key function as effector molecules in apoptosis-related signaling pathways. Any attempt to mediate in such pathways, for example to probe the efficacy or mechanism of action of a drug, would inexorably serve to have a modulatory effect on these proteolytic enzymes. Apoptosis studies of phenanthridone alkaloids of the Amaryllidaceae have only gathered momentum over the past decade, following which caspases have understandably emerged as reliable biochemical markers of the process in an assortment of cancers. This review covers such studies of these alkaloids based on their structural-type, pointing out the various caspases which have been activated in different cancer cells and how structure modification can to a certain extent have a bearing on such activity. Also considered are clues to the apoptosis signaling pathways mediated following phenanthridone-induced activation of caspases.

Several of the Amaryllidaceae alkaloids are known for their cytotoxic properties, of which the lycorine group representatives are prominent for potent and cell line specific antiproliferative activities. As a distinct niche within the lycorine group, the phenanthridones, exemplified by narciclasine and pancratistatin, have shown much promise as remarkably selective cytotoxic agents and are presently at various stages of development, with a clinical candidate likely to appear on the market within the next decade. The crinane group of the Amaryllidaceae has also spawned several molecules, such as crinamine and haemanthamine, with promising cytotoxic activities. In the present study, the β-crinane distichamine as well as the phenanthridone narciprimine, both rare constituents of the Amaryllidaceae, are revealed as novel antiproliferative agents. Apoptosis-inducing effects are demonstrated for distichamine in human acute lymphoblastic leukemia (CEM) cells. These findings provide further insights to the structural details of the apoptosis-inducing pharmacophores resident within both series of alkaloids.

The Amaryllidaceae alkaloids are a group of isoquinoline alkaloids with a distinct array of structural diversity. These compounds have attracted widespread attention due to their manifold biological properties, of which their significance in motor neuron disease through the clinical representative galanthamine is most striking. The cytotoxic potential of Amaryllidaceae alkaloids has also been gaining ground following the advancement of its phenanthridones pancratistatin and narciclasine into clinical trials. This progress has sparked considerable interest in this plant family as a sustainable source of potential anticancer chemotherapeutics. Given this background, this review seeks to (1) examine the cytotoxic effects of Amaryllidaceae alkaloids in cancer cell lines, (2) provide a structural and pharmacophoric basis to these effects, and (3) highlight the various molecular mechanisms behind these activities.

Nine Brazilian Amaryllidaceae species were studied for their alkaloid composition and acetylcholinesterase (AChE) inhibitory activity via GC?MS and a modified Ellman assay, respectively. A total of thirty-six alkaloids were identified in these plants, of which Hippeastrum papilio and H. glaucescens exhibited the highest galanthamine content and the best IC50 values against AChE. Furthermore, Hippeastrum vittatum and Rhodophiala bifida also showed notable AChE inhibitory effects. X-ray crystallographic data for four galanthamine-type compounds revealed significant differences in the orientation of the N-methyl group, which are shown to be related to AChE inhibition. Fil: de Andrade, Jean Paulo. Universidad de Barcelona; España. Universidade Federal do Rio Grande do Sul; Brasil. Universidad del Espiritu Santo; Brasil Fil: Giordani, Raquel B.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Torras-Claveria, Laura. Universidad de Barcelona; España. Universidad del Espiritu Santo; Brasil Fil: Pigni, Natalia Belen. Universidad de Barcelona; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina. Universidade Federal do Rio Grande do Sul; Brasil Fil: Berkov, Strahil. Universidad de Barcelona; España. AgroBioInstitute; Bulgaria. Universidad del Espiritu Santo; Brasil Fil: Font-Bardia, Mercè. Universidad de Barcelona; España Fil: Calvet, Teresa. Universidade Estadual de Campinas; Brasil Fil: Konrath, Eduardo Luis. Universidade Federal do Rio Grande do Sul; Brasil. Universidad del Espiritu Santo; Brasil Fil: Bueno, Kelly. Universidade Federal do Rio Grande do Sul; Brasil Fil: Sachett, Liana G.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Dutilh, Julie H.. Universidade Estadual de Campinas; Brasil. Universidad del Espiritu Santo; Brasil Fil: de Souza Borges, Warley. Universidad del Espiritu Santo; Brasil Fil: Viladomat, Francesc. Universidad de Barcelona; España Fil: Henriques, Amelia T.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Nair, Jerald J.. Universidad de Barcelona; España Fil: Zuanazzi, José Angelo S.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Bastida, Jaume. Universidad de Barcelona; España

Cycads are a group of slow-growing, perennial, dioecious plants which in geological times thrived during the Mesozoic era. The present worldwide complement of around 300 species is distributed through both tropical and subtropical regions of the globe, and has pronounced centres in Australia, Mexico and South Africa. These popular plants are known for their ornamental, economic, dietary and medicinal value. However, increased demand collectively from this sector, as well as decimation of large tracts of cycad populations in the wild through habitat destruction and unscrupulous collection, has precipitated a fragile conservation status for the family, with majority of its species listed as threatened by statutory bodies such as CITES and IUCN. Despite their popularity, cycads have for several centuries been known to be toxic. The plants have been shown to have carcinogenic, mutagenic, teratogenic and neurotoxic properties, and its major constituent principles macrozamin and cycasin have exhibited similar effects in model studies. Cycads are used widely in traditional medicine (TM) in South Africa and the majority of endemic species are commonly traded on local TM markets. The present study deals with the isolation of macrozamin from seed kernels of Encephalartos transvenosus and quantification of this toxic methylazoxymethanol (MAM) glycoside via HPLC in selected indigenous species.

Ethnopharmacological relevance Plants of the family Amaryllidaceae are used widely in traditional medicine in South Africa. Several of these, including representatives of the genus Cyrtanthus find use in the treatment of mental illness and age-related dementia. Aim of the study Based on the distribution of central nervous system-activating alkaloidal constituents within the genus Cyrtanthus , Cyrtanthus contractus was here explored for such compounds which could interact with acetylcholinesterase (AChE), of significance in the progression of neurodegeneration associated with Alzheimer's disease. Materials and methods The known phenanthridone alkaloid narciprimine was isolated via column chromatography of the ethanolic extract of bulbs of Cyrtanthus contractus . The structure of the compound was determined by high field 2D NMR and mass spectroscopic techniques. The classical method of Ellman et al. was used in the determination of AChE inhibitory activity. Results The isolation of narciprimine from Cyrtanthus contractus is a landmark find since it has previously only been identified in Zephyranthes , Narcissus and Lycoris , genera endemic to the Americas, Europe and Asia, respectively. Narciprimine exhibited micromolar inhibitory activity (IC 50 78.9) against the enzyme acetylcholinesterase. Conclusion This work represents the first isolation of narciprimine from an African Amaryllidaceae species, which may be of chemotaxonomic significance. The AChE inhibitory activity of narciprimine, taken together with activities of other structurally similar inhibitors within the series affords further insight to the structural details of the lycorine alkaloid acetylcholinesterase inhibitory pharmacophore.

The synthesis of differentially functionalized analogs of the Amaryllidaceae alkaloid lycorine, accessed via a concise chemoselective silylation strategy, is described uncovering two of the most potent inhibitors of acetylcholinesterase (AChE) identified to date in this series. Important elements of this novel pharmacophore were elucidated through structure-activity relationship (SAR) studies.

seco-Derivatives of the anticancer agent pancratistatin bearing the 2S,3S,4S,5S configuration were accessed via a novel, highly diastereoselective anti-aldol reaction. Structure–activity relationships reveal important insights into the seco-pancratistatin pharmacophore as a potent and selective inhibitor of human cytochrome P450 3A4 (CYP3A4), and highlight features of concern in advancing a potent, selective anticancer agent in the pancratistatin series.

The monodentate ligand 1,3,5,7-tetramethyl-2,4,8-trioxa-6-phenyl-6-phospha-adamantane (PA-Ph) is shown to be highly effective in palladium-catalyzed carbonylative cross-coupling. Aryl and vinyl halides were efficiently converted to carboxylic acids, amides and to primary, secondary, and tertiary esters, respectively. Application of the Pd(OAc)2/PA-Ph (1:1) catalyst system proved critical in the methoxycarbonylation of a functionalized nitroresorcinol halide, allowing convenient access to novel platensimycin sulfonamide analogs.

A library of natural and semi-synthetic Amaryllidaceae alkaloids was screened for cytochrome P450 3A4 (CYP3A4) inhibitory activity. Of the crinane, lycorane and galanthamine representatives examined two semi-synthetic silylated lycorane analogues, accessed via a chemoselective silylation strategy from lycorine, and the natural compound narciclasine exhibited low micromolar activities. Important pharmacological features uncovered include the lack of CYP3A4 inhibitory activity seen for galanthamine and the selective activity that is seen with narciclasine over pancratistatin.

The use of the ligand bis(di-tert-butylphosphino)-o-xylene (dtbpx) in palladium-catalysed carbonylative and Suzuki–Miyaura cross-coupling reactions is described. Aryl and vinyl halides readily entered into the carbonylative catalytic cycle affording carboxylic acids, amides as well as primary, secondary and tertiary esters, respectively, in good yields. Aryl iodides, bromides and chlorides gave high yields of biphenyl products upon reaction with both activated and unactivated boronic acids.

Graphical abstract The direct chemoselective differential functionalization of the ring-C hydroxyl groups present in the Amaryllidaceae alkaloid lycorine is described allowing for selective manipulation of the 1,2-hydroxyl groups. A mini-library comprised of synthetic and natural lycorane alkaloids was prepared and their apoptosis-inducing activity investigated in human leukemia (Jurkat) cells. Further insights into the nature of this interesting apoptosis-inducing pharmacophore are described, including the requirement of both free hydroxyl groups in ring-C., The direct chemoselective differential functionalization of the ring-C hydroxyl groups present in the Amaryllidaceae alkaloid lycorine is described allowing for selective manipulation of the 1,2-hydroxyl groups. A mini-library comprised of synthetic and natural lycorane alkaloids was prepared and their apoptosis-inducing activity investigated in human leukemia (Jurkat) cells. Further insights into the nature of this interesting apoptosis-inducing pharmacophore are described, including the requirement of both free hydroxyl groups in ring-C.

The magnesium chloride catalyzed anti-aldol reaction of phenyl acetate derived oxazolidinones proceeds readily with enolizable L-threose derivative 8 to provide anti-aldol adducts in high yields and with very high diastereoselectivities. The reaction is also efficient with aromatic aldehydes and provides slightly lower diastereoselectivities. This extension

The Amaryllidaceae occupies a privileged status amongst medicinal plants in having delivered the Alzheimer's drug galanthamine to the clinical market. Following its resounding success, there have been several positive indicators for the emergence of an anticancer drug from the family due to the potent antiproliferative activities manifested by several of its alkaloid constituents. Of these, the phenanthridones such as pancratistatin hold most promise as potential chemotherapeutics having succumbed to various phases of clinical trials. Other cytotoxic targets of the Amaryllidaceae are to be found within the lycorane and crinane groups, as exemplified by crinine and lycorine. Although the molecular targets of these alkaloids still remain elusive, much effort has gone into understanding their mode of action in cancer cells. Recent findings have shown that the apoptotic pathway may be a key factor in cancer cell death instigated by Amaryllidaceae alkaloids. As such, this review seeks to: (a) examine the apoptotic effects of Amaryllidaceae alkaloids in cancer cells; (b) explore the molecular basis to these effects; and (c) provide a pharmacophoric rationale in support of these activities.

The [Rh(acac)(CO)(L)] (acac = acetylacetonato; L1 = 1,3-bis-(2,6-diisopropylphenyl)imidazolinylidene and L2 = 1,3-bis-(2,4,6-trimethylphenyl)imidazolinylidene) complexes were prepared by the action of the parent carbene on [Rh(acac)(CO)2] in THF. The crystal structure characterisation of [Rh(acac)(CO)(L1)] revealed a slightly distorted square planar geometry with the carbene ligand orientated almost perpendicular to the equatorial plane; an elongated trans Rh–O bond of 2.0806(18) A reflecting the considerable trans-influence of the carbene ligand. By measuring the CO stretching frequencies in a range of [Rh(acac)(CO)(L)] complexes (L = CO, L1, L2, PPh3, PnBu3, P(O–2,4-tBu2–Ph)3) the following electron donating ability series was established: L1 ∼ L2 ∼ PnBu3 > PPh3 > P(O–2,4-tBu2–Ph)3 > CO; indicating the carbenes investigated in this study to have a similar electronic cis-influence as trialkyl phosphines. Both complexes do not display hydroformylation activity towards 1-hexene in the absence of added phosphine or phosphite ligands under the conditions investigated (P = 60; T = 85 °C). In the presence of a phosphine or phosphite ligand the resulting hydroformylation catalysis was identical to that observed for [Rh(acac)(CO)2] and the corresponding ligand and subsequent high-pressure 31P NMR studies confirmed substitution of the carbene ligand under these conditions.

The novel compounds N-demethylbelladine, 6α-methoxybuphanidrine and filifoline, in addition to five known alkaloids, and phenol have been isolated from fresh bulbs of Nerine filifolia (Amaryllidaceae). The structure and stereochemistry of the compounds were determined by physical and spectroscopic methods, including 1D and 2D NMR and mass spectroscopic techniques. An unusual circular dichroism response from filifoline has required a semi-synthetic derivatisation strategy towards key C-11endo analogues of the β-crinane representative ambelline in which the nature of substituents was observed to have a profound effect on molecular ellipticity. Filifoline was not cytotoxic to myoblast (L6) cells and exhibited no anti-protozoal activity in an in vitro screen against four different parasitic protozoa.

The plant family Amaryllidaceae is known for its ornamental and medicinal value, as well as its unique alkaloid constituents, including several molecules of high biological interest, such as galanthamine, pancratistatin and lycorine. These plants are also used in traditional medicinal practice systems across the globe, and the taxonomy of the family is relatively well-understood. The African genus Boophone Herb. comprises two species, B. disticha (L.f.) Herb. and B. haemanthoides F.M. Leight. Much is known about the ethnobotanical, phytochemical and pharmacological properties of the wide-ranging B. disticha. In contrast, B. haemanthoides is a threatened and territorially restricted species, and its ethnobotanical usage by Khoi-San of the Northern Cape has only recently been described. During our investigation, B. haemanthoides was shown to be a novel source of the known β-crinane alkaloids distichamine, buphanidrine, buphanisine, and crinine. Of note is the presence of distichamine in B. haemanthoides, previously identified only in B. disticha, and its significance as a distinctive chemotaxonomic marker for the genus Boophone.

A catalyst system consisting of a chromium source, a 2,6-disubstituted phenol and an aromatic ether solvent, on activation with triethylaluminium, is active and selective for the trimerisation of ethylene to form 1-hexene. The aromatic ether appears to play a role as both solvent and reagent and is pivotal in ensuring both the good activity of the system as well as selectivity to the desired product, 1-hexene.

The cytochrome P45O activities of the naturally occurring Amaryllidaceae alkaloid narciclasine (3), isolated from Narcissus pseudonarcissus, and synthetic derivative trans-dihydronarciclasine (5) are reported. While narciclasine was found to possess potent inhibitory activity to human CYP3A4, its dihydro analogue was inactive. This study revealed that the C1-C10b double bond is required for inhibition of this crucial metabolizing enzyme. Compound 5 also demonstrated no inhibition of the related human cytochromes CYP19 and CYP1A1. This study elevates the status of trans-dihydronarciclasine (5) as a highly privileged, readily available molecule, with potent and selective anticancer activity.

We determined whether the expression of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) in murine macrophages is regulated by the novel synthetic bacterial lipopeptide JBT 3002. Multilamellar liposomes (MLV) encapsulating JBT 3002 (MLV-JBT 3002) stimulated the production of 72-kDa and 92-kDa (gelatinase A and B) type IV collagenase and inhibited the production of murine metalloelastase (MME) in a dose-dependent manner in murine peritoneal macrophages. MLV-JBT 3002 also induced production of TIMP-1. MLV-JBT 3002 did not induce collagenase production in tumor cells. Priming murine macrophages with interferon-gamma (IFN-gamma) inhibited JBT 3002-stimulated production of both MMP-9 and MMP-2 and further inhibited production of MME by a mechanism involving nitric oxide (NO). This conclusion is based on data showing that IFN-gamma failed to inhibit production of MMP in the presence of L-methyl arginine or in macrophages from inducible nitric oxide synthase knockout mice. These data suggest that JBT 3002 differentially regulates the production of various MMPs and TIMP in macrophages.

Cell membranes contain numerous species of glycosphingolipids (GSL) and glycerophospholipids. Changes in the surface expression of these moieties that accompany malignant transformation are potential targets for immune therapies. There is increasing evidence that tumour specificity resides in the carbohydrate composition and topography of gangliosides and in the asymmetric distribution of phospholipids in the bilayer membrane. However, since these epitopes are not antigenic, circumvention of immunological tolerance to these ‘self antigens’ is required to generate an effective immune response. New methodologies accomplish this by coupling these hapten-like lipid epitopes to immunogenic protein carriers. Indeed, the generation of immune responses to endogenous lipid antigens occurs in several autoimmune syndromes and may serve as natural models for therapeutic immune intervention that influence pathways critical to tumour angiogenesis, proliferation and apoptosis.

Aim of this study was to develop and characterize an applicable in vivo model to investigate angiogenesis of human gliomas. An established glioblastoma spheroid model was used to investigate the neovascularization of a standardized avascular solid tumor mass. Spheroids of two human glioma cell lines were labeled with an in vivo fluorescent dye. Single spheroids were implanted into the cortex of athymic rats. After 1, 3, 7, 14, and 21 days, brain sections containing the spheroid were immunostained for endothelial cells or vascular endothelial growth factor (VEGF). The dye-stained glioma spheroid and the endothelial cells were visualized by confocal microscopy. Two distinct mechanisms of tumor vascularization could be observed. (1) "Classical" angiogenesis with new vessels sprouting from existing host vessels into the spheroid was seen. (2) Individual endothelial cells were found to migrate towards and into the center of the spheroid where they coalesced to form new vessels. This process occurred as early as 24 hr after spheroid implantation. Spheroid vascularization was accompanied by an increase of VEGF expression, which peaked 7 days after implantation and returned to normal patterns by 14-21 days. Besides the "classical" angiogenesis by angiogenic blood vessels, the recruitment of individual endothelial cells seems to be an additional mechanism in early glioma vascularization. Our model proves to be a reliable, reproducible system to study in vivo angiogenesis of human gliomas.

Six alkaloids have been isolated from fresh bulbs of Crinum delagoense . The alkaloids, delagoensine and delagoenine, are reported here for the first time. The structure and stereochemistry of the new alkaloids have been determined by physical and spectroscopic methods. 1 H and 13 C NMR spectra were completely assigned by means of 2D NMR techniques.

Beta-2-glycoprotein I (beta2GPI), a 50-kDA serum glycoprotein that binds negatively charged phospholipids plays a role in coagulation, thrombosis, and the clearance of phosphatidylserine expressing cells. Because of its recently recognized role in several autoimmune responses, we have developed a method that quantifies plasma beta2GPI levels by using a competitive ELISA assay. When combined with data from a standard ELISA, this method determines the concentration of free beta2GPI and the fraction of antibody-bound beta2GPI thereby facilitating quantification of total antigen in individuals with autoimmune antibodies. Standard competitive inhibition ELISA was compared with this method, which uses known amounts of standard beta2GPI added to the plasma as an internal standard. Identical results were obtained with both methods for plasma samples from normal individuals that did not contain blocking antibodies. Analysis of plasma from antiphospholipid syndrome patients (patients with autoantibodies to beta2GPI) by the internal standard method, however, resulted in significantly lower apparent beta2GPI levels indicating that a substantial fraction of the plasma beta2GPI was bound by antibody.

We studied activation to the tumoricidal state of murine peritoneal macrophages by liposomes containing a new synthetic analogue, JBT3002, of a lipoprotein from the outer wall of a gram-negative bacterium. The liposomes containing JBT3002 or CGP31362 were superior to liposomes containing muramyl tripeptide phosphatidylethanolamine (MTP-PE) for tumoricidal activation in three ways. First, efficient macrophage activation required lower concentrations of JBT3002 or CGP31362 than MTP-PE. Second, macrophage activation by JBT3002 was less dependent on priming by interferon-gamma. Third, MLV-JBT3002 activated tumoricidal properties in both lipopolysaccharide (LPS)-responsive and LPS-nonresponsive macrophages. The activation of tumoricidal properties by MLV-JBP3002 depended on protein tyrosine kinase (PTK) activity associated with phosphorylation of tyrosine. The major mechanism for tumoricidal activity in macrophages incubated with MLV-JBT3002 was due to increased activity of inducible nitric oxide synthase (iNOS) and, hence, production of nitric oxide (NO). We base this conclusion on the results of several experiments. First, MLV-JBT3002 was not directly toxic to tumor target cells. Second, the specific iNOS inhibitor NG-monomethyl-L-arginine abrogated tumor cell lysis by MLV-JBT3002-treated macrophages. Third, macrophages from iNOS knockout mice did not lyse tumor cells, even after incubation with high concentrations of MLV-JBT3002. These data suggest that liposomes containing the synthetic bacterial lipopeptide JBT3002 are potent activators of macrophage tumoricidal properties.

Bioisosteric replacement of cyclic ketone functionality with aryl halides was investigated on a centrally-flexible, five-component 1,2,3-triazole-containing pharmacophore, resulting in enhanced inhibition of aromatase (CYP450 19A1). Structure-activity data generated from both syn- and anti-aldol precursors provides significant insights into the requirements for enhanced potency, validating this novel ketone-to-aryl halide bioisostere hypothesis.

The plant family Amaryllidaceae is of provenance in the South African region which is known to harbor about a third of the global complement of around 1000 species. It has widespread usage in the traditional medicinal practices of the indigenous peoples of the region. As a consequence and given its unique alkaloid principles, its members have provided a viable platform for phytochemical based drug discovery. The medicinal potential of the family has been realized through the commercialization of galanthamine as an Alzheimer's drug due to its potent and selective inhibitory activity against the enzyme acetylcholinesterase. Further promising chemotherapeutic candidates of the family reside with the phenanthridone class of alkaloids such as pancratistatin which exhibit potent and cell line specific antiproliferative properties with significant potential for clinical development. Despite these interesting medicinal attributes, plants of the Amaryllidaceae are known to be poisonous and several of them have been classified as such. This survey taking into consideration Amaryllidaceae plants native to South Africa aims to strike a balance between the medicinal potential of the family on one hand and its adverse and toxic effects on the other.

A hexane extract of the wood of Dysoxylum muelleri has yielded 3α-cinnamoyl glabretal, 3α-benzoyl glabretal, 21,24-epoxy-23α,25-dihydroxy-4α,4β,8β-trimethyl-14,18-cyclo-5α,13α,14α,17α-cholestan-3α-cinnamoyl-7α-yl acetate, 21,24-epoxy,23α,25-dihydroxy-4α,4β,8β-trimethyl-14,18-cyclo-5α,13α,14α,17α-cholestan-3α-benzoyl-7α-yl acetate; 21,23-epoxy-21,24,25-trihydroxy-4α,4β,8β-trimethyl-14,18-cyclo-5α,13α,14α,17α-cholestan-3α-cinnamoyl-7α-yl acetate and 21,23-epoxy-21,24,25-trihydroxy-4α,4β,8β-trimethyl-14,18-cyclo-5α,13α,14α,17α-cholestan-3α-benzoyl-7α-yl acetate. A hexane extract of the bark yielded 6α-acetoxy-obacunone acetate.

Purpose: These studies were designed to develop an orthotopic model for human bladder cancer and to isolate variant metastatic cell lines.Materials and Methods: The human bladder cancer cell line 253J was implanted into the muscular wall of the bladder of athymic nude mice. By in vivo recycling, we selected for 2 variant cell lines: 253J B-V, a bladder line isolated after 5 serial passages in the bladder, and 253J lung-IV, established from a lung tumor nodule that was recycled through the bladder.Results: These 2 cell lines showed enhanced tumorigenicity, as measured by a decreased latent period, and rapid growth as compared with the parental cell line. Moreover, orthotopic implantation of these cell lines resulted in metastasis to the lungs. These in vivo-selected, metastatic cell lines exhibited unique karyotypic alterations, increased anchorage-independent growth, overexpression of basic fibroblast growth factor, altered expression of adhesion molecules and the ability to migrate through Matrig...

Turraea floribunda bark has previously been shown to contain limonoids. We now report on the isolation of three new limonoids, turraflorin A, B and C, from the seed of this plant which are ring A, C and D intact, ring B opened, and contain a Δ8,30, Δ14 -diene system.

The discovery of a novel five-component 1,2,3-triazole-containing pharmacophore that exhibits potent and selective inhibition of aromatase (CYP 450 19A1) is described. All compounds are derived from an initial aldol reaction of a phenylacetate derivative with an aromatic aldehyde. Structure–activity data generated from both syn- and anti-aldol adducts provides initial insights into the requirements for both potency and selectivity.

A series of saccharin derivatives I has been synthesized and evaluated for their inhibitory activity toward human leukocyte elastase and cathepsin G. Most of the compounds were found to be efficient and time-dependent inhibitors of elastase. Inactivated elastase was found to regain its activity almost fully after 24 h (80-90% activity) and the half-lives of reactivation ranged between 12-15 h. Addition of hydroxylamine to fully-inactivated enzyme led to rapid and complete recovery of enzymatic activity. A tentative mechanism of action is proposed on the basis of biochemical and model studies.

A series of 3-( alkylthio )-N- hydroxysuccinimide derivatives was synthesized and their inhibitory activity towards human leukocyte elastase (HLE) was investigated. The interaction of the compounds having a 3-alkylthioether side chain (compounds 1 and 2) with HLE was found to involve rapid acylation of the enzyme, followed by total regain of enzymatic activity within 3 h. Interestingly, compounds 3–8, having an oxidized thioether side chain, were found to be highly effective, time-dependent, irreversible inhibitors of the enzyme. The k obs /I values for compounds 3–8 ranged between 890 and 24 000 M−1 s−1. These findings demonstrate that, unlike the physiological inhibitor of HLE (alpha-1-proteinase inhibitor), which is inactivated upon oxidation, low-molecular-weight compounds retain and/or show enhanced inhibitory activity towards HLE upon oxidation of the thioether side chain and lay the groundwork for the development of compounds that embody proteinase inhibitory and antioxidant activity.

Neutrophil-derived mediators such as, for example, the serine proteinase elastase, cathepsin G and proteinase 3, play a critical role in inflammatory lung disease. This report describes the design, synthesis and in vitro inhibitory activity of some novel mechanism-based inhibitors of human leukocyte elastase and cathepsin G. The design of the inhibitors is based on the Gabriel-Colman rearrangement. The behavior of the synthesized compounds toward elastase and cathepsin G with respect to inhibitory prowess, mode of interaction, specificity, etc., has been found to be dependent on the recognition and reactivity elements present in each inhibitor.

A fluorimetric acetylcholinesterase (AChE) assay was developed and characterized both in solution and with the enzyme entrapped in sol-gel-derived silica. The assay is based on a disulfide-thiol interchange reaction between the intramolecularly quenched dimeric dye BODIPY FL l-cystine and thiocholine generated by the AChE-catalyzed hydrolysis of acetylthiocholine (ATCh), which results in a brightly fluorescent monomeric product owing to the cleavage of the disulfide-coupled form of the dye. The new assay was validated by comparison with the Ellman assay performed under parallel conditions and was used in both kinetic and end point assays. The assay was extended to the fabrication of functional AChE microarrays using contact pin-printing of sol-gel-derived silica. A total of 392 sol-gel formulations were screened for gelation times and 192 of these were further evaluated for array fabrication on four different surfaces using a factor analysis approach. Of these, 66 sol-gel/surface combinations produced robust microarrays, while 26 sol-gel/surface combinations were identified that could produce highly active AChE microarrays. The Z' factor for the on-array assay using an optimal sol-gel/surface combination, which considers both signal variability and difference in signals between positive and negative controls, was determined to be 0.60, which is above the minimum level required for applicability to screening. By overprinting nanoliter volumes of solutions containing the dye, ATCh, and potential inhibitors, these microarrays could be used to screen two libraries of small molecules, one composed of newly synthesized alkaloids and another consisting of ∼1000 known bioactive compounds, both as discrete compounds and mixtures thereof, for activity against AChE. IC(50) values were obtained on microarrays for compounds showing significant inhibitory activity, demonstrating the utility of arrays for quantitative inhibition assays.

A general approach toward the rational design of potential inhibitors of serine proteinases is described. The approach is exemplified and validated through the use of appropriate heterocyclic systems in inhibiting human leukocytes elastase (HLE).

The design, synthesis, and inhibitory activity toward human neutropil elastase, of a series of potential mechanims-based inhibitors is described. The design, synthesis, and inhibitory activity toward human neutrophile elastase, of a series of potential mechanism-based inhibitor's (I-III) is described.