Amaryllidaceae Species Research Articles

Chemical Composition of Bioactive Alkaloid Extracts from Some Narcissus Species and Varieties and their Biological Activity

Alkaloid extracts of eight Narcissus (Amaryllidaceae) species and varieties were studied with respect to their acetylcholinesterase (HuAChE) and butyrylcholinesterase (HuBuChE) inhibitory activity and alkaloid patterns. Thirty alkaloids were determined by GC/MS, and twenty-five of them identified from their mass spectra, retention times and retention indexes. Promising HuAChE inhibition activity was demonstrated by six Narcissus taxa and HuBuChE inhibition by N. jonquila cv. Double Campernelle and N. nanus cv. Elka with IC50 values of 24.1 +/- 1.9 microg/mL and 25.1 +/- 1.8 microg/mL, respectively. Two alkaloids were isolated in pure form using preparative TLC and identified as the galanthamine type alkaloid narwedine and the lycorine type alkaloid incartine. Both compounds were tested for their biological activity. They were considered inactive in HuAChE/HuBuChE assays, but showed promising prolyl oligopeptidase inhibition activities with IC50 values of 0.95 +/- 0.12 mM and 0.91 ğ 0.09 mM, respectively.

Synthesis of pyrrolophenanthridine alkaloids based on C(sp³)-H and C(sp²)-H functionalization reactions.

Assoanine, pratosine, hippadine, and dehydroanhydrolycorine belong to the pyrrolophenanthridine family of alkaloids, which are isolated from plants of the Amaryllidaceae species. Structurally, these alkaloids are characterized by a tetracyclic skeleton that contains a biaryl moiety and an indole core, and compounds belonging to this class have received considerable interest from researchers in a number of fields because of their biological properties and the challenges associated with their synthesis. Herein, a strategy for the total synthesis of these alkaloids by using C-H activation chemistry is described. The tetracyclic skeleton was constructed in a stepwise manner by C(sp(3))-H functionalization followed by a Catellani reaction, including C(sp(2))-H functionalization. A one-pot reaction involving both C(sp(3))-H and C(sp(2))-H functionalization was also attempted. This newly developed strategy is suitable for the facile preparation of various analogues because it uses simple starting materials and does not require protecting groups.

Crinine-type alkaloids from Hippeastrum aulicum and H. calyptratum

An ongoing search for alkaloids in the Amaryllidaceae species using GC–MS resulted in the identification of two crinine-type alkaloids, aulicine (1) and 3-O-methyl-epimacowine, (2) from the indigenous Brazilian species Hippeastrum aulicum and Hippeastrum calyptratum, respectively. In addition, two alkaloids, 11-oxohaemanthamine (3) and 7-methoxy-O-methyllycorenine (4) were both isolated from H. aulicum. Furthermore, we provide here complete NMR spectroscopic data for the homolycorine analogues nerinine (5) and albomaculine (6). The absolute stereochemistry of the 5,10b-ethano bridge in the crinine variants was determined by circular dichroism and X-ray crystallographic analysis, thus presenting the first direct evidence for the presence of crinine-type alkaloids in the genus Hippeastrum.

Crinane Alkaloids of the Amaryllidaceae with Cytotoxic Effects in Human Cervical Adenocarcinoma (HeLa) Cells

The family Amaryllidaceae has a long history of usage in the traditional medicinal practices of the indigenous peoples of South Africa, with three of its species known to be used for cancer treatment. Furthermore, the Amaryllidaceae is widely recognized for its unique alkaloid constituents, several of which exhibit potent and selective cytotoxic activities. In this study, several crinane alkaloids derived from local Amaryllidaceae species were examined for cytotoxic effects against the human cervical adenocarcinoma cell line, of which distichamine was the most potent (IC50 2.2 microM).

Smoke-water enhances in vitro pollen germination and tube elongation of three species of Amaryllidaceae

Smoke-water prepared from burning plant material and smoke-derived compounds significantly promote seed germination and enhance growth of many plant species. Since large amounts of smoke are generated and released into the air during wildfires, it is possible that angiosperm pollen germination and pollen tube elongation may be affected by plant-derived smoke even when the plants are some distance from the fire. We assessed the effect of smoke on pollen germination and pollen tube elongation for three species of Amaryllidaceae that occur naturally in areas prone to winter fire in South Africa. In vitro pollen germination and pollen tube growth of Clivia gardenii, Cyrtanthus mackenii and Scadoxus multiflorus were assessed by preparing hanging drop slides with different concentrations of smoke-water, karrikinolide and 3,4,5-trimethylfuran-2(5H)-one combined with Brewbaker and Kwack's medium and a sucrose and boric acid medium. These slides were incubated for 2h at 25°C. Pollen germination and pollen tube lengths were recorded by capturing images with a compound microscope aided by a digital camera. Low concentrations of smoke-water (1:1000 and 1:2000 v:v) significantly increased pollen germination and pollen tube length in the three species, when applied alone or in combination with either Brewbaker and Kwack's medium or sucrose and boric acid medium. Low concentrations of smoke-water significantly increased in vitro pollen germination and pollen tube growth even when no additional calcium was added. Consequently, smoke from grassland fires may have favourable implications for the reproductive process of flowering plants.

Isolation and Cholinesterase Inhibitory Activity of Narcissus Extracts and Amaryllidaceae Alkaloid

Alkaloid extracts of eleven Narcissus (Amaryllidaceae) species and varieties were studied with respect to their acetylcholinesterase (HuAChE) and butyrylcholinesterase (HuBuChE) inhibitory activity and alkaloid patterns. Forty-two alkaloids were determined by GC/MS, and thirty of them identified from their mass spectra, retention times and retention indexes. Promising HuAChE inhibition activity was demonstrated by nine Narcissus taxa and HuBuChE inhibition by Narcissus poeticus cv. Pink Parasol with an IC50 value of 3.3 ± 0.5 μg/mL. The alkaloid identified as (11C- S)-homolycorine was isolated in pure form from Narcissus Sir W. Churchill using preparative TLC and was tested for its biological activity. Homolycorine inhibited HuAChE and HuBuChE in a dose-dependent manner with IC50 values of 63.7±4.3 μM and 151.0±15.2 μM, respectively.

Towards a Molecular Understanding of the Biosynthesis of Amaryllidaceae Alkaloids in Support of Their Expanding Medical Use

The alkaloids characteristically produced by the subfamily Amaryllidoideae of the Amaryllidaceae, bulbous plant species that include well know genera such as Narcissus (daffodils) and Galanthus (snowdrops), are a source of new pharmaceutical compounds. Presently, only the Amaryllidaceae alkaloid galanthamine, an acetylcholinesterase inhibitor used to treat symptoms of Alzheimer’s disease, is produced commercially as a drug from cultivated plants. However, several Amaryllidaceae alkaloids have shown great promise as anti-cancer drugs, but their further clinical development is restricted by their limited commercial availability. Amaryllidaceae species have a long history of cultivation and breeding as ornamental bulbs, and phytochemical research has focussed on the diversity in alkaloid content and composition. In contrast to the available pharmacological and phytochemical data, ecological, physiological and molecular aspects of the Amaryllidaceae and their alkaloids are much less explored and the identity of the alkaloid biosynthetic genes is presently unknown. An improved molecular understanding of Amaryllidaceae alkaloid biosynthesis would greatly benefit the rational design of breeding programs to produce cultivars optimised for the production of pharmaceutical compounds and enable biotechnology based approaches.

QUANTITATIVE DETERMINATION OF GALANTHAMINE AND LYCORINE IN GALANTHUS FOSTERI

s. PHYTOPHARM 2012 M 66 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 QUANTITATIVE DETERMINATION OF GALANTHAMINE AND LYCORINE IN GALANTHUS FOSTERI © Kaya G. I., Emir A, Sarikaya B., Onur M. A., Unver Somer N. Department of Pharmacognosy, Faculty of Pharmacy, Ege University, Bornova Izmir 35100, Turkey Among the 14 taxa and one hybrid of the genus Galanthus growing in Turkey, Galanthus fosteri baker is distributed mainly in south-and north-central Turkey (1, 2). Galanthamine, the most important alkaloid found in Amaryllidaceae species, is used for the treatmant of Alzheimer’s disease (3). lycorine, a widespread alkaloid found in Amaryllidaceae plants has important biological activities (4). Therefore it is important to quantify these alkaloids in Amaryllidaceae species. in the present study, a reversed-phase high-performance liquid chromatographic method has been used for the quantitative determination of lycorine and galanthamine in G. fosteri. The chromatographic separation was performed using an isocratic system with a mobil phase trifluoroacetic acid-water-acetonitrile (0.01: 92.5: 7.5) and diode array dedector (5). The linearity of the method was studied by injecting five known concentrations of lycorine in the range of 2.5–40 μg/ml and five known concentrations of galanthamine in the range of 1–16 μg/ml. The calibration curves for lycorine and galanthamine were determined as y = 15.6024x+0.4459 and y = 10.8302x-1.9292, respectively. The contents of galanthamine in the aerial parts was found to be 0.003 % whereas the amount of the lycorine was found as 0.010 %. The contents of galanthamine and lycorine in the bulbs of G. fosteri were found to be 0.012 % and 0.035 %, respectively.

Alkaloids from Some Amaryllidaceae Species and Their Cholinesterase Activity

Alkaloid extracts of four Amaryllidaceae species were studied with respect to their acetylcholinesterase and butyrylcholinesterase inhibitory activity and alkaloid patterns. Twenty-one alkaloids were determined by GC/MS, and seventeen of them identified from their mass spectra and retention times. The GC/MS analysis of the alkaloid extract of Nerine filamentosa is the first phytochemical investigation of this species. Promising erythrocytic acetylcholinesterase inhibitory activity was demonstrated by the alkaloid extracts of Narcissus poeticus var recurvus, Nerine filifolia and N. filamentosa (IC(50,HuAChE) = 6.0 +/- 0.1 microg/mL; IC(50,HuAChE) = 18.5 +/- 0.8 microg/mL, IC(50,HuAChE) = 21.6 +/- 1.1 microg/mL). The most potent inhibitory activity against serum butyrylcholinesterase was shown by extracts of Sternbergia lutea and Nerinefilamentosa (IC(50,HuBuChE) = 3.7 +/- 0.1 microg/mL; IC(50.HuBuChE) = 13.0 +/- 0.7 microg/mL).

The inhibitory effect of tropolone and hinokitiol on the mycelium growth of Phoma narcissi in vitro

Tropolone and hinokitiol (β-thujaplicin) that are present in heartwood of several Cupressaceae trees are known for their antibacterial, antifungal and insecticidal properties. In the present studies it was showed that tropolone and hinokitiol greatly inhibited <i>in vitro</i>, on PDA medium, the mycelium growth of <i>Phoma narcissi</i>, a pathogen of <i>Hippeastrum</i> and other species of family Amaryllidaceae. Total inhibition of the mycelium growth of <i>Phoma narcissi</i> took place at a tropolone concentration of 6.0 µg·cm<sup>-3</sup> and at a hinokitiol concentration of 50.0 µg·cm<sup>-3</sup>. Fungicidal doses of tropolone and hinokitiol for the mycelium growth of <i>Phoma narcissi</i> were also documented. The results presented in this paper are discussed with data available in literature on the antifungal action of tropolone and hinokitiol on other species of pathogenic fungi.

GC/MS Analysis of Three Amaryllidaceae Species and Their Cholinesterase Activity

Alzheimer's disease (AD), characterized by the death of nerve cells in the cerebral cortex, is the most common subtype of dementia. Despite the exponential growth in the number of AD patients, acetylcholinesterase (AChE) inhibitors are currently used to treat AD. Plants of the Amaryllidaceae family are known to synthesize a particular type of bioactive compounds, named Amaryllidaceae alkaloids, which have shown AChE inhibitory activity. Alkaloid extracts of three species of Amaryllidaceae were studied with respect to their acetylcholinesterase and butyrylcholinesterase inhibitory activity and alkaloid patterns. Eleven alkaloids were identified by GC/MS. Significant cholinesterase inhibitory activity was demonstrated by the alkaloid extract of N. undulata (IC50,(HuAChE) = 14.3 +/- 1.2 microg/mL; IC50,(HuBuChE) = 33.9 +/- 1.9 microg/mL).

Quantitative Determination of Galanthamine and Lycorine in an endemic Galanthus species: G. cilicicus

Galanthus cilicicus Baker, an endemic species of the genus Galanthus L. (Amaryllidaceae), is distributed in southern Turkey mainly in the province of Içel [1]. Galanthamine, the most important alkaloid found in Amaryllidaceae species, is used for the treatment of Alzheimer's disease [2]. Lycorine, another important and also a widespread alkaloid found in Amaryllidaceae plants has been proven to have several biological activities [3,4]. A reversed-phase high-performance liquid chromatographic method has been used and validated for the determination of lycorine and galanthamine in G. cilicicus. The extraction of both alkaloids in low-mass plant samples, were carried out by a simple and a rapid method utilizing pre-packed columns with diatomaceous earth (Extrelut®) [5]. The chromatographic separation was performed using an isocratic system with a mobil phase of trifluoroacetic acid-water-acetonitrile (0.01: 92.5: 7.5) and diode array dedector [6]. The linearity of the method was studied by injecting five known concentrations of lycorine in the range of 1–10µg mL-1 and five known concentrations of galanthamine in the range of 2.5–20µg mL-1 The calibration curves for lycorine and galanthamine were determined as Y=13.2828995x + 0.4488635 and Y=10.1354031x +0.5465348, respectively. Validation procedures showed that the method was specific, accurate and precise. The above-mentioned method was applied to the aerial parts and bulbs of G.cilicicus. The contents of galanthamine and lycorine in the bulbs of G.cilicicus were found to be 0.016% and 0.0035%, respectively. In the aerial parts, lycorine was not detected, however the content of galanthamine was found as 0.015%.

Isolation of narciprimine from Cyrtanthus contractus (Amaryllidaceae) and evaluation of its acetylcholinesterase inhibitory activity

Ethnopharmacological relevancePlants 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 studyBased 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 methodsThe 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. ResultsThe 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 (IC50 78.9) against the enzyme acetylcholinesterase. ConclusionThis 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.

Effects of sucrose and plant growth regulators on acetylcholinesterase inhibitory activity of alkaloids accumulated in shoot cultures of Amaryllidaceae

The influence of sucrose (30, 60, 90 and 120 g/L), activated charcoal (5 and 10 g/L), and various levels of several plant growth regulators (6-benzyladenine, naphthalene-1-acetic acid, 2,4-dichlorophenoxyacetic acid, and picloram) on organogenesis (bulb and root development) and the accumulation of alkaloid and galanthamine in shoot cultures of three Amaryllidaceae species (Narcissus pseudonarcissus, Galanthus elwesii, and Leucojum aestivum) was investigated in a full-factorial experiment. Alkaloid extracts were analyzed by gas chromatography–mass spectrometry, leading to the quantification of galanthamine and to the identification of other alkaloids. The different extracts were then subjected to an Ellman test to evaluate the inhibitory activity of acetylcholinesterase. The highest contents of galanthamine [0.02–0.1% dry weight (DW) depending on the plant species] were always accompanied with a high level of acetylcholinesterase inhibition (>30%). However, some samples containing low amounts of galanthamine (0.005% DW) showed high inhibitory activities (40–80%). These findings demonstrate the presence of Amaryllidaceae alkaloids that have not yet been identified as having anti-acetylcholinesterase activity.

Screening and Bioguided Fractionation of Amaryllidaceae Species with Anti-Trichomonas vaginalisActivity

The Amaryllidaceae family is known by its ornamental and medicinal value and has attracted considerable attention due to the content of alkaloids of its species, which showed interesting biological properties. The present study evaluated the anti-Trichomonas vaginalis activity of eighteen extracts (12.5 to 0.19 mg/mL) and six isolated alkaloids (125 to 1.9 µg/mL) from Amaryllidaceae species. The alkaloids diminished the trophozoites viability (from 15 to 40 %). The extracts from Hippeastrum breviflorum demonstrated the highest anti-T. vaginalis activity (viability was 60 % reduced), and a bioguided study was conducted. Six fractions with antiprotozoal activity had lycorine and lycosinine as major components suggesting a synergistic effect, taking into account the higher anti-T. vaginalis activity of extracts when compared to isolated alkaloids. Our results point out the antiprotozoal potential of the Amaryllidaceae species against T. vaginalis. This parasite causes trichomonosis, the most prevalent nonviral sexually transmitted diseases (STD) worldwide and a public health problem that requires new therapeutic alternatives as well bioactive natural products.

Screening of plants of Amaryllidaceae and related families from Panama as sources of acetylcholinesterase inhibitors

Context: This is the first comprehensive study of the acetylcholinesterase (AChE) inhibitory activity of species of the family Amaryllidaceae and 13 related families from Panama.Objective: Exploration of the potential sources of AChE inhibitors with radical scavenging activity from Amaryllidaceae and 13 related families from Panama.Materials and methods: The studied plants were screened with anti-acetylcholinesterase bioautographic and DPPH free radical scavenging assays.Results: From the 57 plants studied, eight (14%) showed strong inhibition of AChE, and 29 (51%) plants showed moderate inhibition of AChE.Discussion and conclusion: Sagittaria lancifolia L. (Alismataceae), Crinum jagus (Thomps.) (Amaryllidaceae), Crinum x amabile Donn (Amaryllidaceae), Crinum zeylanicum (L.) L. (Amaryllidaceae), Crocosmia x crocosmiiflora (Lemoine ex Anonymous) N.E. Br. (Iridaceae), Sisyrinchium tinctorium Kunth (Iridaceae), Agapanthus praecox subsp. orientalis (F.M. Leight.) F.M. Leight. (Liliaceae), and Xyris jupicai Rich. (Xyridaceae) were the most active plants, inhibiting AChE at 100 μg on the TLC bioautographic method for the detection of acetylcholinesterase inhibitors. Out of the eight most active plants, two plants, Crinum zeylanicum (L.) L. and Xyris jupicai Rich., showed antioxidant activity.

Isolation of acetylcholinesterase inhibitory alkaloids from Nerine bowdenii

Alzheimer's disease (AD) is associated with a steady loss of attention and memory, which has been correlated with the impairment of brain cholinergic neurotransmission, particularly a deficit of cholinergic neurons in the nucleus basalis of Meynert. Therefore, one of the methods that has proven successful in the treatment of AD is the use of inhibitors of acetylcholinesterase (AchE) to supplement the acetylcholine level. Herein we describe the isolation of several AchE-inhibiting alkaloids from the bulbs of Nerine bowdenii, an Amaryllidaceae species. For the first time 11-O-acetylambelline and filofiline, previously reported from Brunsvigia josephinea and Nerine filifolia, respectively, were isolated.

A família Amaryllidaceae no Parque Estadual do Ibitipoca, Minas Gerais, Brasil

The Ibitipoca State Park (Parque Estadual do Ibitipoca - PEIB), in southeastern Minas Gerais, Municipality of Lima Duarte, has an area of 1923.5 hectares, at altitudes varying between 1.100 and 1.784 m. Its vegetation is a mosaic of Atlantic forest andcampo rupestre. Within the studied area three species of Amaryllidaceae were found: Habranthus irwinianus, Hippeastrum aulicum and Hippeastrum glaucescens. The species are found in campo rupestre, on the rocks, over 1600 m elevation. Identification key, descriptions, illustrations and comments are presented.

Quantitative determination of lycorine in Amaryllidaceae plants

Lycorine, a widespread alkaloid in Amaryllidaceae species, has been proven to have antiviral, cytotoxic and antimalarial activities [1–3]. A reversed-phase high-performance liquid chromatographic method has been developed and validated for the determination of lycorine in Amaryllidaceae plants. A simple method for the extraction of lycorine in low-mass plant samples was employed utilizing pre-packed columns with diatomaceous earth (Extrelut®) [4]. The chromatographic separation was performed using an isocratic system with a mobile phase trifluoroacetic acid-water-acetonitrile (0.01:90:10) and diode array detector [5]. The linearity of the method was studied by injecting five known concentrations of the standard in the range of 62.5–500µgmL-1. The calibration curve was determined as Y=23.9788285 X + 62.391901. Validation procedures showed that the method was specific, accurate and precise.

Alkaloids from Boophone disticha with affinity to the serotonin transporter

Boophone disticha L. Herb (Amaryllidaceae) is used in traditional medicine for treatment of painful wounds, headaches, skin disorders, inflammatory conditions, rheumatic pains and anxiety. At least eight alkaloids have been characterized and reported in the literature. Of these buphanidrine and buphanamine have affinity to the serotonin transporter (SERT). Alkaloids from other Amaryllidaceae species have also shown affinity to SERT. In this study, an ethanol extract was prepared from dry bulbs. Through HPLC–UV separation five peaks were collected and characterized by LC-MS and 1H NMR and led to the identification of crinine, buphanamine, buphanidrine, distichamine and buphanisine. The activity of these compounds was tested in a binding assay using [3H]-citalopram as ligand and a functional SERT inhibition assay utilizing COS-7 cells expressing hSERT. The four active compounds, buphanamine, buphanidrine, buphanisine and distichamine, had IC50-values of 55 µM, 62 µM, 199 µM and 65 µM respectively, in the binding assay. The alkaloids also showed activity in the functional assay, buphanidrine and distichamine being the most active with IC50-values of 513 µM and 646 µM, respectively.