Narcissus Bulbs Research Articles

Narciclasine ameliorated T cell mediated acute liver injury through activating AMPK pathway

Hepatitis is closely related to cirrhosis and liver cancer, and it is vital that we develop new drugs and identify new drug targets. Traditional Chinese medicine has demonstrated excellent curative effects on liver diseases. The ingredients from Chinese herbals are important source for drug development in the treatment of hepatitis. Here, we found that narciclasine (NCS), a major component extracted from narcissus bulbs, showed hepatoprotective effect against concanavalin A (Con A) induced hepatitis. NCS treatment significantly reduced hepatocyte death, hepatic inflammatory cells infiltration, and serum cytokine levels in Con A challenged mice. We further observed that NCS directly inhibited Con A induced splenocytes proliferation and cytokine production in vitro. RNA-seq results showed that genes related to immune response were upregulated in Con A treated CD4+ T cells, which were down-regulated in the presence of NCS. Moreover, the AMPK pathway had been found activated in response to NCS treatment, suggesting a potential target for NCS targets. In conclusion, our results reveal that NCS is a powerful immunosuppressor against T cell activation, thus leading to protection against Con A induced liver injury in mice. These findings provide new insights into the use of natural products in the treatment of autoimmune hepatitis.

Pressurized Natural Deep Eutectic Solvent Extraction of Galanthamine and Related Alkaloids from Narcissus pseudonarcissus.

The isolation of a compound from a natural source involves many organic and mostly toxic solvents for extraction and purification. Natural deep eutectic solvents have been shown to be efficient options for the extraction of natural products. They have the advantage of being composed of abundantly available common primary metabolites, being nontoxic and environmentally safe solvents. The aim of this study was to develop a natural deep eutectic solvent-based extraction method for galanthamine, an important therapeutic agent for the treatment of Alzheimer's disease. This alkaloid can be produced by synthesis or by extraction from Narcissus bulbs. To develop an efficient extraction method, a number of different natural deep eutectic solvents was first tested for their solubilization capacity of galanthamine bromide salt. Promising results were obtained for ionic liquids, as well as some amphoteric and acidic natural deep eutectic solvents. In a two-cycle extraction process, the best solvents were tested for the extraction of galanthamine from bulbs. The ionic liquids produced poor yields, and the best results were obtained with some acid and sugar mixtures, among which malic acid-sucrose-water (1 : 1 : 5) proved to be the best, showing similar yields to that of the exhaustive Soxhlet extraction with methanol. Furthermore, the natural deep eutectic solvent was more selective for galanthamine.

Complete genome sequence of vallota mosaic virus detected in a narcissus bulb imported from the United States to Japan.

Narcissus (Narcissus albidus) imported from the United States exhibited leaf chlorosis during post-entry quarantine. We employed next-generation sequencing (NGS) on symptomatic leaf samples and detected vallota mosaic virus (ValMV), belonging to the genus Potyvirus, family Potyviridae, as the viral agent. Sanger sequencing of PCR products and rapid amplification of cDNA ends based on NGS contigs revealed that ValMV is 9,451 nucleotides (nt) in length, excluding the poly(A) tail. Nucleotide and amino acid (aa) sequences of the coat protein region had over 98% identity to previously reported ValMV isolates. In each of the 10 regions encoding mature proteins, however, the sequence identity to other potyviruses was 49.5-71.9% nt and 18.3-78.9% aa, values that are below the species demarcation thresholds for the family Potyviridae. Phylogenetic analysis revealed that our ValMV isolate is most closely related to known ValMV isolates and is grouped with other potyviruses. Taken together, our results indicate that the newly isolated ValMV belongs to a distinct species in the genus Potyvirus. This study provides the first report of the complete ValMV genome sequence and the first record of this virus in narcissus.

Galantamine Quantity and Alkaloid Profile in the Bulbs of Narcissus tazetta and daffodil cultivars (Amaryllidaceae) Grown in Israel.

Alkaloids produced by the bulbs of the Amaryllidaceae are a source of pharmaceutical compounds. The main alkaloid, galantamine, is a reversible acetylcholinesterase inhibitor and allosteric nicotinic receptor modulator, which slows cognitive and functional decline in mild to moderate dementia due to Alzheimer’s disease. Having a complex stereochemistry, the organic synthesis of galantamine for pharmaceutical uses is highly challenging and not always economically viable, and it is therefore isolated from Amaryllidaceae bulbs. In the present study, galantamine was extracted and quantified in Narcissus bulbs from five cultivars (cvs.), Fortune, Carlton, Ice Follies, Galilee and Ziva, which were grown in Israel under various conditions. Results show that the cvs. Fortune, Carlton and Ice Follies bulbs contained 285 ± 47, 452 ± 73 and 69 ± 17 µg g−1 galantamine, respectively, while the Galilee and Ziva bulbs contained relatively low concentrations of galantamine (1–20 µg g−1). Irrigation levels and pruning conditions did not affect the galantamine contents. Additionally, the alkaloids profile of the five cvs. was analyzed and characterized using LC-MS/MS showing that galantamine-type alkaloids were mainly detected in the Fortune and Carlton bulbs, lycorine-type alkaloids were mainly detected at the Galilee and Ziva bulbs and vittatine-type alkaloids were mainly detected in the Ice Follies bulbs. The present research is the first to characterize the alkaloids profile in the Narcissus bulbs of Galilee and Ziva, indigenous cvs. grown in Israel. The antiviral and anticancer alkaloids lycorine and lycorinine were the main alkaloids detected in the bulbs of those cultivars.

Effect of bulb age on alkaloid contents of narcissus pseudonarcissus bulbs

Narcissus is mostly grown for horticultural/decorative purposes in the fields and home gardens. Previously, galanthamine isolation (from the bulbs to produce medicine for the treatment of Alzheimer's disease) has been established as a novel use. Moreover, recent studies have also shown that Narcissus bulbs are also a source of several other interesting biologically active compounds and particularly some alkaloids. Various metabolites are produced at different time points during the growth cycle of plants. Also, for Narcissus bulbs it is important to find from an economical point of view the optimum time point for harvesting in its growth cycle, as the levels of galanthamine might be correlated with the age of the Narcissus bulbs. The aim of this study was to investigate the differences in the metabolites and specifically in the alkaloid contents based on a 1, 2- or 3-years harvesting cycle of Narcissus bulbs. Moreover, the effect of the cultivation site was studied for Narcissus pseudonarcissus cv Carlton which is well known for galanthamine production. Different methods were used for the analysis of these alkaloids. Untargeted NMR based metabolomics was used for overall metabolic changes and galanthamine quantitation. Whereas targeted GC-FID and GC–MS were used respectively for quantitative and qualitative analysis of major and minor alkaloids. An alkaloid specific acid-base extraction method was used in the GC–MS and GC–FID analysis. The results show that the contents of the metabolites vary significantly with the age of the bulbs. The contents of galanthamine were highest in two years old bulbs. It was also found that the alkaloid contents from the same variety grown at different locations differ from each other. Results presented here clearly indicate that] age of the bulb are important factors to consider for the industrial production of certain alkaloids from the plants as yields of alkaloids may vary, certain conditions may lead to an increase of the yield with 10–15%.

Characteristics of azole-resistant Aspergillus fumigatus attached to agricultural products imported to Japan

Due to the increase in the number of azole-resistant Aspergillus fumigatus, there is an urgent need of data to predict future trends and prevent further spreading. The intercountry transfer of resistant A. fumigatus on plant bulbs have been reported. We investigated existence and characteristics of resistant isolates attached to agricultural products imported to Japan.We purchased 292 samples in Japan. All samples were screened for the existence of azole-resistant A. fumigatus. For positive isolates, minimum inhibitory concentrations of the drugs were determined. We also analyzed Cyp51A, Hmg1, and Erg6 mutations of these isolates and conducted microsatellite genotyping.Fourteen azole-resistant isolates were detected, of which 13 were cultured from flower bulbs imported from the Netherlands. Among them 5 were from 11 bulbs of Hippeastrum (45.5%), 5 were from 24 bulbs of Gladiolus (20.8%), 2 were from 4 bulbs of Ixia (50.0%), and 1 was from 22 bulbs of Tulipa (4.5%). Only 1 resistant isolate was cultured from the 10 bulbs of Narcissus (10.0%) originating in Japan. Various novel mutations including Y121F/T289A in Cyp51A with no tandem repeat in promoter region were discovered from imported strains.Our study provides important data showing that agricultural imports provide a possible route for their intercontinental spread and raises the concern that strains harboring highly diverse Cyp51A mutations might increase in clinical settings in the future.

A multiplex RT-PCR method for the simultaneous detection of Narcissus yellow stripe virus, Narcissus latent virus and Narcissus mosaic virus

Narcissus (Narcissus spp.) is an important commercial crop and millions of narcissus bulbs are traded across the world annually. Three major viruses, namely, Narcissus yellow stripe virus (NYSV), Narcissus latent virus (NLV) and Narcissus mosaic virus (NMV), have substantial impacts on global crop yields of narcissus. A multiplex RT-PCR assay was developed and used to simultaneously detect these three viruses. Three pairs of virus-specific primers were designed according to the conserved regions of the coat protein (CP) genes, generating expected fragments of 668 bp, 554 bp and 372 bp for NYSV, NLV and NMV, respectively. Specificity and compatibility of primer pairs were tested using uniplex and multiplex RT-PCR. Primer concentration, annealing temperature and cycle number were optimized to enable efficient and balanced amplification of the three viruses. The optimized multiplex RT-PCR was as sensitive as uniplex RT-PCR. Field samples were collected to validate the method, and all the viruses were successfully detected, thereby confirming the effectiveness and reliability of the developed multiplex RT-PCR. This method could be used for large-scale surveys of these three viruses in narcissus and thus might play a role in diagnosis and prevention of these pathogens.

The Frequency of Viral Infections on Two Narcissus Plantations in Central Poland

Abstract Viral diseases in narcissus can drastically affect yields and quality of narcissus bulbs and flowers, leading even to a total crop loss. To test the frequency of viral infections in production fields in Central Poland, samples were collected over three years from two cultivars and two plantations, and tested for the presence of Arabis mosaic (ArMV), Cucumber mosaic (CMV), Narcissus latent (NLV), Narcissus mosaic (NMV) and the potyvirus group using the Enzyme Linked ImmunoSorbent Assay. Potyviruses, NLV and NMV were detected in almost all leaf samples in both cultivars, in all three years of testing. Other viruses were detected in a limited number of samples. In most cases mixed infections were present. Tests on bulbs have shown the presence of potyviruses and NMV, with the higher number of positives in cultivar ‘Carlton’. In addition, for most viruses an increase in their detectability was observed on both plantations in subsequent seasons.

Standards to be withdrawn

Standards to be withdrawn

Effects of fungicides on galanthamine and metabolite profiles in Narcissus bulbs

Large-scale plant cultivation usually involves the use of pesticides. Apart from eliminating the target organism, the external chemicals may affect the metabolism of the crop plant. This may have implications for plants cultivated for specific medicinal compounds. In this study the effects of diverse fungicides on the metabolism of Narcissus pseudonarcissus cv. Carlton bulbs were investigated. N. pseudonarcissus cv. Carlton is being cultivated for the extraction of the alkaloid galanthamine. Fungicides typically used in Narcissus cultivation were applied in a field experiment. The aim was to determine whether fungicide applications changed the concentration of galanthamine in the bulbs. (1)H NMR spectroscopy allowed quantitative analysis of galanthamine and other metabolites in bulb extracts. Multivariate data analysis revealed changes in bulb metabolite patterns caused by fungicides. Bulbs treated before planting generally had higher levels of alkaloids, while foliar field applications caused lower alkaloid levels but altered carbohydrate metabolism. Within these groups, certain fungicide treatments caused changes in specific metabolites. This study shows that the fungicides used in Narcissus cultivation can cause a change in the metabolome still detectable in the bulbs after harvest. The standard cultivation practices in terms of fungicide treatment were found suitable for the production of N.pseudonarcissus cv. Carlton as raw material for galanthamine extraction. In the cultivation of medicinal plants for secondary metabolites the potential effect of pesticides and other agrochemicals should be taken into account.

Effects of fungicides on galanthamine and metabolite profiles in Narcissus bulbs

Effects of fungicides on galanthamine and metabolite profiles in Narcissus bulbs

Salivary gland hypertrophy virus of house flies in Denmark: prevalence, host range, and comparison with a Florida isolate

House flies (Musca domestica) infected with Musca domestica salivary gland hypertrophy virus (MdSGHV) were found in fly populations collected from 12 out of 18 Danish livestock farms that were surveyed in 2007 and 2008. Infection rates ranged from 0.5% to 5% and averaged 1.2%. None of the stable flies (Stomoxys calcitrans), rat-tail maggot flies (Eristalis tenax) or yellow dung flies (Scathophaga stercoraria) collected from MdSGHV-positive farms displayed characteristic salivary gland hypertrophy (SGH). In laboratory transmission tests, SGH symptoms were not observed in stable flies, flesh flies (Sarcophaga bullata), black dump flies (Hydrotaea aenescens), or face flies (Musca autumnalis) that were injected with MdSGHV from Danish house flies. However, in two species (stable fly and black dump fly), virus injection resulted in suppression of ovarian development similar to that observed in infected house flies, and injection of house flies with homogenates prepared from the salivary glands or ovaries of these species resulted in MdSGHV infection of the challenged house flies. Mortality of virus-injected stable flies was the highest among the five species tested. Virulence of Danish and Florida isolates of MdSGHV was similar with three virus delivery protocols, as a liquid food bait (in sucrose, milk, or blood), sprayed onto the flies in a Potter spray tower, or by immersiion in a crude homogenate of infected house flies. The most effective delivery system was immersion in a homogenate of ten infected flies/ml of water, resulting in 56.2% and 49.6% infection of the house flies challenged with the Danish and Florida strains, respectively.

Abstract 3261: Novel hemisynthetic derivatives of Amaryllidaceae alkaloids exert antitumor activity on xenograft melanoma models

Abstract Alkaloids pancratistatin and narciclasine, isolated from Amaryllidaceae, have been shown to present antitumor activity on preclinical models, including melanoma. Despite extensive investigations, pancratistatin or narciclasine analogues did not succeed to provide sufficient therapeutic benefit, [1] mostly hampered by poor water solubility and systemic toxicity [1]. To overcome those limitations, we developed a chemistry program aimed at improving both their physicochemical and pharmacological properties. A literature survey suggests that any structural modification of this highly oxigenated isocarbostiryl specific scaffold would lead to a loss of activity except on position 1, difficult to achieve but interesting to explore. A series of 40 novel 1-aminopancratistatin derivatives was synthesised starting from large quantities of narciclasine extracted from Narcissus bulbs. These novel compounds exhibit an improved pharmaceutical profile facing standard solubility, plasmatic and microsomal stability assays. Moreover, a subset of narciclasine derivatives demonstrated a significant enhancement of the antiproliferative effect with IC50 reaching the nM range, consistent with a major induction of pro-apoptotic signals, including mitochondrial depolarisation. To explore the mode of action of these alkaloids derivatives, we tested their capacity to inhibit protein synthesis, compared their pharmacological profile with 80 anticancer compounds on 16 tumor cell lines (oncoprofile.16) and explored their impact on major cellular functions such as cellular integrity, cell cycle, topoisomerases inhibition, DNA damage and synthesis. We demonstrated thus that inhibition of DNA synthesis is a key component of their mechanism of action. Based on the pharmacological properties, a structure-activity card can be built to select different categories of pancrastistatin derivatives depending on their ability to inhibit preferentially protein synthesis or/and DNA synthesis. Interestingly, the dimethylaminomethyl benzamide derivative F98604 exhibited a significant antiproliferative activity on the murine B16 and human A375 melanoma xenograft models associated with an increased therapeutic index as compared with the natural compound narciclasine. In conclusion, modulation of position 1 on the narciclasine pharmacophore overcomes the solubility issue and is a key position to modify their pharmacological profiles, this opening new perspectives as anticancer agents active against melanoma. 1. A. Kornienko, A. Evidente Chem. Rev. 2008, 108, 1982 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3261. doi:10.1158/1538-7445.AM2011-3261

Effect of Fertilizers on Galanthamine and Metabolite Profiles in Narcissus Bulbs by 1H NMR

Narcissus bulbs contain the biologically active alkaloid galanthamine, and Narcissus is being developed as a natural source of the molecule for the pharmaceutical industry. The effect of fertilizer on galanthamine production was investigated in a field study using a (1)H nuclear magnetic resonance (NMR) metabolite profiling approach. Galanthamine was quantitated and major metabolites in the bulbs were identified. The application of standard fertilization levels of nitrogen and potassium caused a significant increase in galanthamine as compared to a control. Multivariate data analysis of the (1)H NMR data revealed that applying double the standard level of nitrogen fertilizer resulted in production of more amino acids and citric acid cycle intermediates, but not more galanthamine. The results indicated that standard levels of fertilizer currently applied in The Netherlands are sufficient for optimal galanthamine accumulation in the bulbs. This study shows how (1)H NMR-based metabolic profiling can provide insight into the response of plant metabolism to agricultural practices.

Universal primers for rapid detection of hytrosaviruses

Hytrosaviridae is a proposed virus family encompassing viruses that cause salivary gland hypertrophy (SGH) syndrome in infected insects and reduce the fertility in their dipteran insect hosts. They contain a large, double stranded DNA genome of 120–190 kbp. To date, these viruses have been detected only in adult Diptera. These include hytrosaviruses detected in various tsetse fly species ( Glossina spp.), the narcissus bulb fly Merodon equestris and the house fly Musca domestica. The limited number of hytrosaviruses reported to date may be a reflection of the frequent absence of external symptoms in infected adult flies and the fact that the virus does not cause rapid mortality. Based on the complete genome sequence of Glossinia pallidipes (GpSGHV) and Musca domestica (MdSGHV) salivary gland hypertrophy viruses, a PCR based methodology was developed to detect the viruses in these species. To be able to detect hytrosaviruses in other Diptera, five degenerate primer pairs were designed and tested on GpSGHV and MdSGHV DNA using gradient PCR with annealing temperatures from 37 to 61 °C. Two pairs of primers were selected from p74, two pairs from PIF-1 and one pair from ODV-e66 homologous proteins. Four primer pairs generated a virus specific PCR product on both MdSGHV and GpSGHV at all tested annealing temperatures, while the ODV-e66 based primers did not generate a virus specific product with annealing temperatures higher that 47 °C. No non-specific PCR product was found when using genomic DNA of infected flies as template DNA. These results offer new sets of primers that could be used to detect hytrosaviruses in other insects.

Avoidance of suicide in antibiotic-producing microbes

Many microbes synthesize potentially autotoxic antibiotics, mainly as secondary metabolites, against which they need to protect themselves. This is done in various ways, ranging from target-based strategies (i.e. modification of normal drug receptors or de novo synthesis of the latter in drug-resistant form) to the adoption of metabolic shielding and/or efflux strategies that prevent drug-target interactions. These self-defence mechanisms have been studied most intensively in antibiotic-producing prokaryotes, of which the most prolific are the actinomycetes. Only a few documented examples pertain to lower eukaryotes while higher organisms have hardly been addressed in this context. Thus, many plant alkaloids, variously described as herbivore repellents or nitrogen excretion devices, are truly antibiotics-even if toxic to humans. As just one example, bulbs of Narcissus spp. (including the King Alfred daffodil) accumulate narciclasine that binds to the larger subunit of the eukaryotic ribosome and inhibits peptide bond formation. However, ribosomes in the Amaryllidaceae have not been tested for possible resistance to narciclasine and other alkaloids. Clearly, the prevalence of suicide avoidance is likely to extend well beyond the remit of the present article.

Abstract 5461: Novel antitumor amino-pancratistatin derivatives inhibitors of protein and DNA synthesis

Abstract The Amaryllidaceous alkaloids pancratistatin and narciclasine are known to exert marginal antitumor properties but associated with toxicities. Despite extensive investigations, pancratistatin or analogues did not succeed to provide sufficient therapeutic benefit although a new pro-drug strategy was applied recently to narciclasine. We decided to undertake a chemistry program aimed at modifying the skeleton of this attractive pharmacophore in order to improve its therapeutic application. At first sight, literature survey suggests that any change in the structure would lead to a loss of activity but a careful examination of published data indicated that position −1 of the molecule may be of potential interest although poorly investigated. We report here the synthesis of a series of 39 novel derivatives of 1-aminopancratistatin by chemical modification of the naturally available narciclasine extracted from Narcissus bulbs. These new compounds present improved pharmaceutical properties such as higher aqueous solubility (from 300 µg/ml for narciclasine up to 2000 µg/ml) without loss of stability tested on human and murine microsomal assays (70-90% stability after 1h incubation). A sub-set of these novel derivatives demonstrated a drastic increase of the cytotoxic activity reaching nM range tested on a panel of 8 different cell lines. The higher antiproliferative activity translates towards the induction of stronger pro-apoptotic signals evidenced with Annexin V, caspase 3/7 and JC-1 assay. Besides the previously described inhibition of protein synthesis, our studies originally demonstrated that part of the mechanism of action involves inhibition of DNA synthesis as well. The contribution of these two pharmacological properties elaborates a SAR (structure - activity relationship) and segregates our new pancrastistatin derivatives into different categories depending on their ability to inhibit protein synthesis, DNA synthesis or both. Based on their differential in vitro activity profile, the compounds were selected for evaluation of their in vivo antitumor properties on murine and xenografted models. As a major result F 98604 (dimethylaminomethyl benzamide derivative of pancratistatin) exhibited significant and reproducible activity on the murine B16 sc and human A375 melanoma models in terms of tumor growth inhibition as well as increased therapeutic index. The present SAR suggests that modification of the natural pancratistatin or narciclasine pharmacophore at the position −1 open new perspectives as potential anticancer agents with novel pharmacological profiles. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5461.

Temporal and spatial expression patterns of the LEAFY homologue NLF during florogenesis in Narcissus tazetta

Studies of flower development in numerous plants suggest that the LEAFY gene ( LFY) of Arabidopsis and its homologues in other species act as integrators of floral inductive signals and determine the fate of meristems. In Narcissus tazetta, the LFY homologue NLF was identified, and its expression pattern during flower development was studied under various temperature conditions . In N. tazetta, flower initiation and differentiation occur within the bulb during summer dormancy, and are promoted by relatively high temperatures (25–30 °C). NLF was detected in the apical meristem throughout its development from the vegetative state to fully differentiated flowers, under ambient summer conditions in Israel. A dramatic increase in expression was observed twice: during meristem transition from vegetative to reproductive development, and during differentiation of flower primordia. At later stages of flower differentiation, NLF was expressed in the developing anthers. NLF was also found in leaf primordia, but not in mature leaves, suggesting a role in young leaf development. Temperature conditions during florogenesis affected NLF expression in the apical meristem. Summer storage of Narcissus bulbs at a constant 30 °C resulted in florogenesis and an expression pattern of NLF similar to that observed under ambient temperatures. However, at 12 °C, meristems remained morphologically vegetative and did not produce flowers. At the same time, a high expression level was observed in these non-differentiated meristems. Our results suggest that NLF plays a major role in floral development of N. tazetta, but that temperature does not affect NLF expression directly.

Analysis of metabolic variation and galanthamine content in Narcissus bulbs by 1H NMR

Galanthamine is a benzazepine alkaloid used as a drug to relieve symptoms of Alzheimer's disease. For pharmaceutical use this natural product has been extracted from the plant Leucojum aestivum (Amaryllidaceae) or produced synthetically. Limited supply of the natural source and high cost of synthetic production has led to a search for alternative sources of galanthamine. The bulbs of Narcissus pseudonarcissus (Amaryllidaceae) have been identified as a potential source of raw material for galanthamine extraction. Since inconsistent chemical composition can be an issue with medicinal plant material, it is of interest to know whether large variations occur between Narcissus bulbs grown in different geographical locations. To evaluate whether large differences exist in the overall metabolic profiles of Narcissus bulbs grown in the two most important cultivation regions. (1)H NMR and principal component analysis were used for an unbiased comparison of the bulb samples. Overall metabolite profiles were quite similar, but galanthamine levels could slightly discriminate samples by geographical region. (1)H NMR was used for quantitation of galanthamine, and was found to be comparable to quantitation by HPLC. Compared with conventional chromatographic methods, sample preparation for (1)H NMR analysis is simple and rapid, and only a small amount of plant material is required. Since useful qualitative and quantitative information about the metabolic state of Narcissus bulbs can be obtained by (1)H NMR, this method is useful for agricultural applications, and for quality control of raw material used in the pharmaceutical industry.

Chemistry, biology, and medicinal potential of narciclasine and its congeners.

Ornamental flower growers know that placing a cut daffodil (a.k.a. narcissus) in a vase with other flowers has a negative effect on the quality of those flowers and significantly shortens their vase life. Furthermore, a common horticultural practice for the cultivation of narcissus flowers involves the introduction of cuts on the bulbs before immersing them into water. The mucilage that leaches out from the cuts is constantly removed by frequent changing of water and this leads to sprouting. These observations raise speculation that specific components in the mucilage of the narcissus bulbs may have powerful growth-inhibitory effects. Historical use of narcissus flowers, as well as at least thirty other plants of the Amaryllidaceae family, in folk medicine for the management of cancer1 speaks volumes to validate this conjecture. Indeed, powerful anticancer properties of Narcissus poeticus L. were already known to the Father of Medicine, Hippokrates of Kos (ca. B.C. 460–370), who recommended a pessary prepared from narcissus oil for the treatment of uterine tumors.2 His successors, the ancient Greek physicians Pedanius Dioscorides (ca. A.D. 40–90) and Soranus of Ephesus (A.D. 98–138) continued using this therapy in the first and second centuries A.D.3,4 In addition, the topical anticancer uses of extracts from this plant5,6 as well as from N. pseudonarcissus7–9 were recorded in the first century A.D. by the Roman natural philosopher Gaius Plinius Secundus, (A.D. 23–79), better known as Pliny the Elder.10 Even the Bible provides multiple references to the Mediterranean N. tazetta L., which has a long history of use against cancer.11 The applications of narcissus oil in cancer management continued in the middle ages in Chinese, North African, Central American and Arabian medicine.1,12 The uses of other genera of the Amaryllidaceae family were also common, e. g. Hymenocallis caribaea (L. emend Gawler) Herbert, utilized by early European medical practitioners for inflammatory tumors.13 More recently, the plants of the Amaryllidaceae have been under intense scrutiny for the presence of the specific metabolites responsible for the medicinal properties associated with this plant family. The study began in 1877 with the isolation of alkaloid lycorine from Narcissus pseudonarcissus14 and since then more than 100 alkaloids, exhibiting diverse biological activities, have been isolated from the Amaryllidaceae plants. Based on the present scientific evidence, it is likely that isocarbostyril constituents of the Amaryllidaceae, such as narciclasine, pancratistatin and their congeners, are the most important metabolites responsible for the therapeutic benefits of these plants in the folk medical treatment of cancer. Notably, N. poeticus L. used by the ancient Greek physicians, as was eluded before, is now known to contain some 0.12 g of narciclasine per kg of fresh bulbs.15 Continuing along this intriguing path, the focus of the present review is a comprehensive literature survey and discussion of the chemistry and biology of these compounds as specifically relevant to their potential use in medicine. The examination of the synthetic organic chemistry, more specifically the total synthesis efforts inspired by the challenging chemical structures of narciclasine, pancratistatin and their congeners, will be reduced to a minimum in view of the two very recent excellent reviews published on this subject.16,17