RESUMO
Despite extensive and intensive research efforts in recent decades, there is still no effective treatment for neurodegenerative diseases. On this background, the use of drugs inhibiting the enzyme acetylcholinesterase (AChE) remains an eternal evergreen in the symptomatic treatment of mild to moderate cognitive impairments. Even more, the cholinergic hypothesis, somewhat forgotten in recent years due to the shift in focus on amyloid cascade, is back to life, and the search for new, more effective AChE inhibitors continues. We generated a fragment-based library containing aromatic moieties and linkers originating from a set of novel AChE inhibitors. We used this library to design 1220 galantamine (GAL) derivatives following the model GAL (binding core) - linker (L) - aromatic fragment (Ar). The newly designed compounds were screened virtually for blood-brain barrier (BBB) permeability and binding to AChE. Among the top 10 best-scored compounds, a representative lead molecule was selected and tested for anti-AChE activity and neurotoxicity. It was found that the selected compound was a powerful non-toxic AChE inhibitor, 68 times more active than GAL, and could serve as a lead molecule for further optimization and development.
Assuntos
Inibidores da Colinesterase/análise , Desenho de Fármacos , Descoberta de Drogas , Avaliação Pré-Clínica de Medicamentos , Interface Usuário-Computador , Acetilcolinesterase/química , Acetilcolinesterase/metabolismo , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Linhagem Celular , Inibidores da Colinesterase/química , Galantamina/química , Galantamina/farmacologia , Camundongos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Neurotoxinas/toxicidade , Bibliotecas de Moléculas PequenasRESUMO
Acetylcholinesterase (AChE) inhibitors remain the class of drugs used for the treatment of Alzheimer disease (AD). For the aim of discovering new sources of potent AChE inhibitors, a combined AChE-inhibitory activity together with alkaloid profiles by GC-MS, combined with multivariate statistical analysis for biomarkers determination and in silico studies were attempted. Strategy was applied on leaves, roots and bulbs of six aquatic and terrestrial Amaryllidaceae species. Thirty alkaloids were identified and the AChE inhibitory activities of the extracts were tested by in-vitro Ellman method. Principal bioactive markers were discovered by correlating AChE inhibitory activity with chemical fingerprints via PLS and OPLS modeling which revealed that galanthamine, lycoramine, caranine, tazettine and N-demethylgalanthamine were the most bio-significant markers. Furthermore, the molecular docking was performed to illustrate binding orientations of the top scoring alkaloids in the active site of human acetylcholinesterase. Suggested strategy revealed that, beside galanthamine, caranine, N-demethylgalanthamine, and lycoramine are promising AChE inhibitors.
Assuntos
Alcaloides/farmacologia , Amaryllidaceae/química , Inibidores da Colinesterase/farmacologia , Simulação por Computador , Crinum/química , Cromatografia Gasosa-Espectrometria de Massas , Acetilcolinesterase/metabolismo , Alcaloides/química , Doença de Alzheimer , Alcaloides de Amaryllidaceae/química , Alcaloides de Amaryllidaceae/farmacologia , Domínio Catalítico , Inibidores da Colinesterase/química , Galantamina/química , Galantamina/farmacologia , Humanos , Simulação de Acoplamento Molecular , Análise Multivariada , Extratos Vegetais/química , Folhas de Planta/química , Raízes de Plantas/químicaRESUMO
Plants of the Amaryllidaceae family are promising therapeutic tools for human diseases and have been used as alternative medicines. The specific secondary metabolites of this plant family, called Amaryllidaceae alkaloids (AA), have attracted considerable attention due to their interesting pharmacological activities. One of them, galantamine, is already used in the therapy of Alzheimer's disease as a long acting, selective, reversible inhibitor of acetylcholinesterase. One group of AA is the montanine-type, such as montanine, pancracine and others, which share a 5,11-methanomorphanthridine core. So far, only 14 montanine-type alkaloids have been isolated. Compared with other structural-types of AA, montanine-type alkaloids are predominantly present in plants in low concentrations, but some of them display promising biological properties, especially in vitro cytotoxic activity against different cancerous cell lines. The present review aims to summarize comprehensively the research that has been published on the Amaryllidaceae alkaloids of montanine-type.
Assuntos
Alcaloides de Amaryllidaceae/química , Amaryllidaceae/química , Antineoplásicos Fitogênicos/química , Antiprotozoários/química , Inibidores da Colinesterase/química , Nootrópicos/química , Amaryllidaceae/metabolismo , Alcaloides de Amaryllidaceae/isolamento & purificação , Alcaloides de Amaryllidaceae/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Antiprotozoários/isolamento & purificação , Antiprotozoários/farmacologia , Linhagem Celular Tumoral , Inibidores da Colinesterase/isolamento & purificação , Inibidores da Colinesterase/farmacologia , Galantamina/química , Galantamina/isolamento & purificação , Galantamina/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/química , Compostos Heterocíclicos de 4 ou mais Anéis/isolamento & purificação , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Humanos , Concentração Inibidora 50 , Isoquinolinas/química , Isoquinolinas/isolamento & purificação , Isoquinolinas/farmacologia , Nootrópicos/isolamento & purificação , Nootrópicos/farmacologia , Fenantridinas/química , Fenantridinas/isolamento & purificação , Fenantridinas/farmacologia , Extratos Vegetais/química , Metabolismo SecundárioRESUMO
Narcissus tazetta is used traditionally for treatment of sores, wounds, skin diseases, cancer in different parts of world. Present study focus on the analysis of amaryllidaceae alkaloids in this plant using an ultra-performance liquid chromatography-diode array detection method. The method was developed for simultaneous quantification of eight Amaryllidaceae alkaloids i.e. pseudolycorine (1), lycorine (2), galanthamine (3), 8-O-demethylhomolycorine (4), N-methylhaemanthidine chloride (5), homolycorine (6), narciclasine (7) and zefbetaine (8) in Narcissus tazetta. The method was validated using a BEH C18 column with linear gradient. Standard calibration curve for the analytes showed good linearity (â¯r2≥0.999). The method was validated for intra-day (RSDs<0.91%) and inter-day (RSDs<0.65%) precisions and accuracy (recovery 92.2-112.5%). The developed method was successively applied for studying the variation of alkaloids in different parts of Narcissus tazetta, i.e. bulbs, roots, flowers, flower stalks and leaves. The study showed a significant variation of these alkaloids in different parts of the plant. Among the alkaloids under investigation, pseudolycorine had highest content in all the parts. Furthermore, application of the developed method to the identification of phytocomponents allowed the identification of sixteen alkaloids.
Assuntos
Alcaloides de Amaryllidaceae/química , Narcissus/química , Extratos Vegetais/química , Cromatografia Líquida de Alta Pressão/métodos , Flores/química , Galantamina/química , Fenantridinas/química , Folhas de Planta/química , Raízes de Plantas/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodosRESUMO
Narcissus spp. are an economically important crop for medicines in relation with the alkaloids production, mainly galanthamine, an acetylcholinesterase inhibitor used for the treatment of Alzheimer's disease. In this article an extensively study of the phytochemistry of both bulbs of different species and varieties of Narcissus grown in Iran and in vitro culture of these plants was investigated. In particular, the Amaryllidaceae alkaloid profile and the galanthamine and lycorine contents in wild bulbs of Narcissus papyraceus (G5) and four varieties of Narcissus tazetta (N. tazetta var. Shahla (G4), N. tazetta var. Shastpar (G1), N. tazetta var. Meskin (G2), N. tazetta var. Panjehgorbei (G3)), growing in Iran are reported. The alkaloid profiles were investigated by GC-MS and LC-MS and the quantitative analysis was performed using GC-MS. In total, thirty alkaloids were identified among them nine alkaloids were observed with the both methods of analysis. The variety Meskin of N. tazetta (G2), showed the highest diversity of alkaloids and the highest content in galanthamine. On this last species (G2) and on N. tazetta var. Shahla (G4), the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (Picloram) and naphthalene acetic acid (NAA) at concentrations of 25 and 50 µM were studied on the induction of callus and its capacity to induce organogenesis and alkaloid diversity. All auxins, at the concentrations of 25 and 50 µM, produced calli. Bulblets and roots were formed on calli grown only in the presence of 25 or 50 µM NAA. GC-MS analyses showed the presence of galanthamine and lycorine in calli, roots and bulblets, with all auxins whatever the concentration used while demethylmaritidine and tazettine were found in differentiated tissue cultures cultivated on the medium containing NAA (25 or 50 µM) or in calli initiated with Picloram (50 µM). Precursor 4'-O-methylnorbelladine (MN) of Amaryllidaceae alkaloids feeding was found to significantly improve the accumulation of both galanthamine (82 µg/g DW) and lycorine (1800 µg/g DW) in bulblets of N. tazetta var. Meskin (G2).
Assuntos
Alcaloides de Amaryllidaceae/química , Narcissus/química , Inibidores da Colinesterase/química , Galantamina/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Irã (Geográfico) , Fenantridinas/química , Extratos Vegetais/química , Raízes de Plantas/químicaRESUMO
A virtual screening protocol based on machine learning models was used to identify mimetics of the natural product (-)-galantamine. This fully automated approach identified eight compounds with bioactivities on at least one of the macromolecular targets of (-)-galantamine, with different polypharmacological profiles. Two of the computer-generated hits possess an expanded spectrum of bioactivity on targets relevant to the treatment of Alzheimer's disease and are suitable for hit-to-lead expansion. These results advocate multitarget drug design by advanced virtual screening protocols based on chemically informed machine learning models.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Produtos Biológicos/farmacologia , Inibidores da Colinesterase/farmacologia , Desenho de Fármacos , Galantamina/farmacologia , Aprendizado de Máquina , Fármacos Neuroprotetores/farmacologia , Acetilcolinesterase/metabolismo , Doença de Alzheimer/metabolismo , Produtos Biológicos/síntese química , Produtos Biológicos/química , Linhagem Celular Tumoral , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Avaliação Pré-Clínica de Medicamentos , Galantamina/síntese química , Galantamina/química , Humanos , Ligantes , Simulação de Acoplamento Molecular , Estrutura Molecular , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , EstereoisomerismoRESUMO
Acetylcholinesterase (AChE) catalyzes the hydrolysis of neurotransmitter acetylcholine to acetate and choline in a synaptic cleft. Deficits in cholinergic neurotransmitters are linked closely with the progression of Alzheimer's disease (AD), which is a neurodegenerative disorder characterized by memory impairment, and a disordered cognitive function. Since the previously approved AChE inhibitors, donepezil (Aricept), galantamine (Reminyl), and rivastigmine (Exelon), have side effects and several studies are being carried out out to develop novel AD drugs, we have applied a three-dimensional quantitative structure-activity relationship (3D QSAR) and structure-based pharmacophore modeling methodologies to identify potential candidate inhibitors against AChE. Herein, 3D QSAR and structure-based pharmacophore models were built from known inhibitors and crystal structures of human AChE in complex with donepezil, galantamine, huperzine A, and huprine W, respectively. The generated models were used as 3D queries to screen new scaffolds from various chemical databases. The hit compounds obtained from the virtual screening were subjected to an assessment of drug-like properties, followed by molecular docking. The final hit compounds were selected based on binding modes and molecular interactions in the active site of the enzyme. Furthermore, molecular dynamics simulations for AChE in complex with the final hits were performed to evaluate that they maintained stable interactions with the active site residues. The binding free energies of the final hits were also calculated using molecular mechanics/Poisson-Boltzmann surface area method. Taken together, we proposed that these hits can be promising candidates for anti-AD drugs.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Inibidores da Colinesterase/química , Inibidores da Colinesterase/uso terapêutico , Descoberta de Drogas , Acetilcolinesterase/metabolismo , Alcaloides/química , Sítios de Ligação , Bases de Dados de Compostos Químicos , Donepezila/química , Galantamina/química , Humanos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Ligação Proteica , Relação Quantitativa Estrutura-Atividade , Sesquiterpenos/químicaRESUMO
Sixteen new alkaloids belonging to the galanthamine (1-6), plicamine (7-14), and secoplicamine (15 and 16) classes, together with eight known analogues (17-24), were isolated from Zephyranthes candida. The structures of 1-16 were determined by extensive spectroscopic analyses, and the absolute configurations of 1, 2, 7, 8, and 17 were confirmed by single-crystal X-ray diffraction analysis. The orientation of 3-OCH3 in N-methyl-5,6-dihydroplicane (22) was revised. Alkaloids 3, 12-14, and 18-21 exhibited anti-acetylcholinesterase activities with IC50 values ranging from 0.48 to 168.7 µM. Compounds 10-12, 14, and 16 showed in vitro anti-inflammatory activities with IC50 values ranging from 7.50 to 23.55 µM.
Assuntos
Anti-Inflamatórios não Esteroides/isolamento & purificação , Inibidores da Colinesterase/isolamento & purificação , Inibidores da Colinesterase/farmacologia , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Galantamina/isolamento & purificação , Galantamina/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/isolamento & purificação , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Liliaceae/química , Plantas Medicinais/química , Acetilcolinesterase/metabolismo , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Inibidores da Colinesterase/química , Cristalografia por Raios X , Medicamentos de Ervas Chinesas/química , Galantamina/química , Compostos Heterocíclicos de 4 ou mais Anéis/química , Concentração Inibidora 50 , Medicina Tradicional , Conformação Molecular , Estrutura Molecular , Ressonância Magnética Nuclear BiomolecularRESUMO
Melatonin is a neurohormone whose levels are significantly reduced or absent in Alzheimer's disease (AD) patients. In these patients, acetylcholinesterase inhibitors (AChEI) are the major drug class used for their treatment; however, they present unwanted cholinergic side effects and have provided limited efficacy in clinic. Because combination therapy is being extensively used to treat different pathological diseases such as cancer or acquired immune deficiency syndrome, we posed this study to evaluate if melatonin in combination with an AChEI, galantamine, could provide beneficial properties in a novel in vitro model of AD. Thus, we subjected organotypic hippocampal cultures (OHCs) to subtoxic concentrations of ß-amyloid (0.5 µM ßA) plus okadaic acid (1 nM OA), for 4 days. This treatment increased by 95 % cell death, which was mainly apoptotic as shown by positive TUNEL staining. In addition, the combination of ßA/OA increased Thioflavin S aggregates, hyperphosphorylation of Tau, oxidative stress (increased DCFDA fluorescence), and neuroinflammation (increased IL-1ß and TNFα). Under these experimental conditions, melatonin (1-1000 nM) and galantamine (10-1000 nM), co-incubated with the toxic stimuli, caused a concentration-dependent neuroprotection; maximal neuroprotective effect was achieved at 1 µM of melatonin and galantamine. Most effective was the finding that combination of sub-effective concentrations of melatonin (1 nM) and galantamine (10 nM) provided a synergic anti-apoptotic effect and reduction of most of the AD-related pathological hallmarks observed in the ßA/OA model. Therefore, we suggest that supplementation of melatonin in combination with lower doses of AChEIs could be an interesting strategy for AD patients.
Assuntos
Doença de Alzheimer/patologia , Galantamina/farmacologia , Hipocampo/patologia , Melatonina/farmacologia , Técnicas de Cultura de Tecidos , Peptídeos beta-Amiloides/toxicidade , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Morte Celular/efeitos dos fármacos , Galantamina/química , Melatonina/química , Modelos Biológicos , Fármacos Neuroprotetores/farmacologia , Ácido Okadáico/toxicidade , Fosforilação/efeitos dos fármacos , Agregados Proteicos/efeitos dos fármacos , Ratos Sprague-Dawley , Proteínas tau/metabolismoRESUMO
OBJECTIVE: The amyloid hypothesis stimulates the discovery of compounds, which promotes beta-amyloid peptide (Aß) clearance, thereby altering the underlying pathophysiology of Alzheimer's disease (AD). Hence, the present study aims at the evaluation of anti-amyloidogenic potential of Gelidiella acerosa. METHODS: Prevention of Aß 25-35 aggregate formation and disaggregation of pre-formed fibrils by G. acerosa was evaluated in three phases by thioflavin T spectrophotometric assay. The results were further validated by confocal microscopic analysis. The conformational changes in the aggregated and non-aggregated Aß in the presence of G. acerosa were analyzed by Fourier transform infrared (FTIR) spectroscopic analysis. RESULTS: Phase-I study shows that G. acerosa reverts (4.56 ± 0.35 AU at 96 hours) the increase in fluorescence intensity of aggregated Aß (18.76 ± 0.99 AU) significantly (P < 0.05) as that of non-aggregated peptides, which suggests that G. acerosa prevents the formation of oligomers from monomers. The seaweed also prevents the fibril formation even after the aggregation process was initiated at 20 hours, which was verified by the significant (P < 0.05) decrease in the fluorescence intensity (2.94 ± 0.0721 AU) at 36 hours (Phase II). In addition, G. acerosa promotes fibrillar destabilization (Phase III), which was further substantiated by confocal microscopic analysis. Fourier transform infrared spectroscopy reveals that alteration in amide I and amide II band spectrum, which occurs due to Aß 25-35 aggregation was restored upon co-treatment with G. acerosa benzene extract. CONCLUSION: Overall, the results suggest that G. acerosa might have direct interaction with the aggregated peptide, thereby preventing oligomerization and fibrillation of Aß 25-35.
Assuntos
Peptídeos beta-Amiloides/metabolismo , Fármacos Neuroprotetores/farmacologia , Fragmentos de Peptídeos/metabolismo , Extratos Vegetais/farmacologia , Rodófitas , Alga Marinha , Amiloide/metabolismo , Benzeno/química , Fracionamento Químico , Galantamina/química , Microscopia Confocal , Fármacos Neuroprotetores/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Multimerização Proteica/efeitos dos fármacos , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The phytochemistry of Kopsia arborea Blume has received considerable attention, which has resulted in the isolation of a number of new unusual indole alkaloids with intriguing structures. In this study, a new eburnane-type alkaloid, phutdonginin (1), together with eight known alkaloids: 19-OH-(-)- eburnamonine (2), melodinine E (3), kopsinine (4), kopsilongine (5), kopsamine (6), (-)-methylenedioxy-1 1,12-kopsinaline (7), decarbomethoxykopsiline (8), and vincadifformine (9), were isolated from the twigs of K. arborea. Their structures were characterized extensively by 1D and 2D NMR spectroscopy and HR-ESI-MS. All compounds were submitted to TLC screening for acetylcholinesterase inhibitory activities. Only kopsamine and decarbomethoxykopsiline showed AChE inhibition with MIR values of 12.5 and 6.25 µg, respectively, compared with galanthamine (positive control, 0.004 µg). In addition, compounds 1 and 2 inhibited moderate antibacterial activity against E. coli TISTR 780 with the MIC value of 32 .g/mL.
Assuntos
Alcaloides/química , Monoterpenos/química , Plantas Medicinais/química , Alcaloides/farmacologia , Antibacterianos/química , Antibacterianos/farmacologia , Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Cromatografia em Camada Fina , Escherichia coli/efeitos dos fármacos , Galantamina/química , Galantamina/farmacologia , Alcaloides Indólicos/química , Alcaloides Indólicos/farmacologia , Espectroscopia de Ressonância Magnética , Alcaloides de Triptamina e Secologanina/química , Alcaloides de Triptamina e Secologanina/farmacologia , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
In order to investigate the effects of phenylalanine, tyrosine and tyramine on the growth of Lycoris radiata suspension cells and the accumulation of alkaloids, the growth quantity of the cells as well as the content of alkaloids in cells were determined, which were treated with above three kinds of precursors alone and phenylalanine combined with tyrosine respectively. The results indicate that the addition of phenylalanine alone and addition of phenylalanine on the basis of tyrosine at high concentration (200 micromol/L) had no significant effect on the growth of Lycoris radiata suspension cells and the content of alkaloids in cells; whereas tyrosine and tyramine promoted the growth of the cells and alkaloids accumulation. Treated with tyrosine at high concentration (200 micromol/L), the content of alkaloids of the cells was 2.56-fold higher than that of the control group, the amounts of lycoramine (3.77 mg/g) and galanthamine (4.46 mg/g) were 6.61-fold and 6.97-fold higher than that of the control group, respectively. When treated with tyramine (200 micromol/L), the amount of alkaloids in Lycoris radiata suspension cells was 2.63-fold higher than that of the control group, and the amounts of lycoramine (4.45 mg/g) and galanthamine (5.14 mg/g) were 9.08-fold and 9.18-fold higher than that of the control group, respectively. The above results demonstrate that adding tyrosine and tyramine in the media significantly promoted the growth of the Lycoris radiata suspension cells and alkaloids accumulation in the cells.
Assuntos
Alcaloides de Amaryllidaceae/química , Lycoris/crescimento & desenvolvimento , Células Vegetais/química , Células Cultivadas , Meios de Cultura/química , Galantamina/química , Lycoris/química , Fenilalanina/química , Extratos VegetaisRESUMO
The etiopathology of Alzheimer's disease (AD) is extremely complex and heterogeneous, often associated with comorbidities. As a result it may be unlikely that AD may be mitigated by drug acting on a single specific target. The current tendency in drug design and discovery in AD is the rational design or "serendipitous" discovery of new drug entities challenging multiple targets. Since two of the presently approved drugs for AD are based on natural products (galantamine and the physostigmine-derivative rivastigmine), many plants are now under investigation as a potential source of new drugs. Multifunctional drugs often have their origin in natural sources. This review is limited to plant chemicals having different targets with actual (galantamine) or promising (drugs from Crocus sativus, Ginkgo biloba, Salvia species, and Huperzia serrata) clinical evidence in people with dementia or AD.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Produtos Biológicos/farmacologia , Descoberta de Drogas/métodos , Extratos Vegetais/farmacologia , Plantas/química , Alcaloides/química , Alcaloides/farmacologia , Animais , Produtos Biológicos/química , Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Crocus/química , Galantamina/química , Galantamina/farmacologia , Galanthus/química , Ginkgo biloba/química , Humanos , Huperzia/química , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Fisostigmina/química , Fisostigmina/farmacologia , Extratos Vegetais/química , Salvia/química , Sesquiterpenos/química , Sesquiterpenos/farmacologiaRESUMO
The Amaryllidaceae family is well known for its pharmacologically active alkaloids. An important approach to treat Alzheimer's disease involves the inhibition of the enzyme acetylcholinesterase (AChE). Galanthamine, an Amaryllidaceae alkaloid, is an effective, selective, reversible, and competitive AchE inhibitor. This work was aimed at studying the alkaloid composition of four wild Argentinian Amarillydaceae species for the first time, as well as analyzing their inhibitory activity on acetylcholinesterase. Alkaloid content was characterized by means of GC-MS analysis. Chloroform basic extracts from Habranthus jamesonii, Phycella herbertiana, Rhodophiala mendocina and Zephyranthes filifolia collected in the Argentinian Andean region all contained galanthamine, and showed a strong AChE inhibitory activity (IC50 between 1.2 and 2 µg/mL). To our knowledge, no previous reports on alkaloid profiles and AChEIs activity of wild Argentinian Amarillydaceae species have been publisihed. The demand for renewable sources of industrial products like galanthamine and the need to protect plant biodiversity creates an opportunity for Argentinian farmers to produce such crops.
Assuntos
Alcaloides/química , Inibidores da Colinesterase/química , Galantamina/química , Liliaceae/química , Extratos Vegetais/química , Alcaloides/isolamento & purificação , Argentina , Inibidores da Colinesterase/isolamento & purificação , Cromatografia em Camada Fina , Galantamina/isolamento & purificação , Cromatografia Gasosa-Espectrometria de Massas , Extratos Vegetais/isolamento & purificação , Raízes de Plantas/químicaRESUMO
A rapid, simple and sensitive ultra performance liquid chromatography-diode array detection method (UPLC-DAD) was developed and validated for quantification of four biologically important Amaryllidaceae alkaloids viz. lycoramine, hamayne, haemanthamine and tortuosine in Zephyranthes grandiflora. The method employed BEH C(18) column (2.1mm×100mm, 1.7µm particle size) with linear gradient elution of acetonitrile and water (0.05% formic acid) in a flow rate of 0.3mL/min and at λ(max) 280nm. Standard calibration curve for the analytes were linear (r(2)≥0.9999), precise (intra-day RSDs<1.33% and inter-day RSDs<2.67%) and accurate (97.8-105.3%). The developed method was applied to the quality assessment of samples collected during different seasons and showed significant variation with an optimum amounts of these alkaloids in rainy season. The method was also applied for identification of Amaryllidaceae alkaloids in the plant and overall, seventeen Amaryllidaceae alkaloids of different structural types lycorine, haemanthamine, galanthamine, narciclasine were characterised. This study provides a qualitative and quantitative method for analysis of Amaryllidaceae alkaloids.
Assuntos
Alcaloides de Amaryllidaceae/química , Cromatografia Líquida de Alta Pressão/métodos , Liliaceae/química , Extratos Vegetais/química , Calibragem , Galantamina/química , Fenantridinas/química , Pirróis/químicaRESUMO
The bulbs and aerial parts of Zephyranthes concolor (Lindl.) Benth. & Hook. f. (Amaryllidaceae), an endemic species to Mexico, were found to contain the alkaloids chlidanthine, galanthamine, galanthamine N-oxide, lycorine, galwesine, and epinorgalanthamine. Since currently only partial and low resolution (1)H-NMR data for chlidanthine acetate are available, and none for chlidanthine, its 1D and 2D high resolution (1)H- and (13)C-NMR spectra were recorded. Unambiguous assignations were achieved with HMBC, and HSQC experiments, and its structure was corroborated by X-ray diffraction. Minimum energy conformation for structures of chlidanthine, and its positional isomer galanthamine, were calculated by molecular modelling. Galanthamine is a well known acetylcholinesterase inhibitor; therefore, the isolated alkaloids were tested for this activity. Chlidanthine and galanthamine N-oxide inhibited electric eel acetylcholinesterase (2.4 and 2.6 × 10(-5) M, respectively), indicating they are about five times less potent than galanthamine, while galwesine was inactive at 10(-3) M. Inhibitory activity of HIV-1 replication, and cytotoxicity of the isolated alkaloids were evaluated in human MT-4 cells; however, the alkaloids showed poor activity as compared with standard anti-HIV drugs, but most of them were not cytotoxic.
Assuntos
Acetilcolinesterase/metabolismo , Alcaloides/química , Inibidores da Colinesterase/química , Liliaceae/química , Extratos Vegetais/química , Alcaloides/metabolismo , Alcaloides/farmacologia , Alcaloides de Amaryllidaceae/química , Alcaloides de Amaryllidaceae/metabolismo , Alcaloides de Amaryllidaceae/farmacologia , Animais , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacologia , Linhagem Celular , Inibidores da Colinesterase/metabolismo , Inibidores da Colinesterase/farmacologia , Electrophorus , Galantamina/química , Galantamina/metabolismo , Galantamina/farmacologia , HIV-1/efeitos dos fármacos , Humanos , Ligação de Hidrogênio , Liliaceae/anatomia & histologia , Dados de Sequência Molecular , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Difração de Raios XRESUMO
Seventy alkaloids of galanthamine, lycorine, homolycorine, tazettine, haemanthamine, narciclasine, and tyramine types were detected by GC/MS in 25 Galanthus elwesii and seven Galanthus nivalis populations, collected from different locations in Bulgaria. Intraspecies diversity in the alkaloid profiles regarding the main alkaloid types (chemotypes) was observed. Tyramine-type protoalkaloids (namely, hordenine and its derivatives) were dominant in 19 populations of G. elwesii. In other populations of G. elwesii, the plants accumulated mainly homolycorine-, lycorine-, and galanthamine-type alkaloids. The alkaloid profiles of G. nivalis were dominated by narciclasine-, galanthamine-, lycorine-, haemanthamine-, or tazettine-type compounds. Geographical distribution of chemotypes indicated a relationship between populations, since adjacent populations often displayed similar alkaloid profiles. The results from year-to-year sampling and transplantation experiments imply genetic determination of alkaloid synthesis in the two studied species of Galanthus.
Assuntos
Alcaloides/química , Galanthus/química , Alcaloides de Amaryllidaceae/química , Galantamina/química , Cromatografia Gasosa-Espectrometria de Massas , Fenantridinas/químicaRESUMO
Galanthamine is an Amaryllidaceae-type alkaloid with acetylcholinesterase inhibitory activity which is used in the treatment of Alzheimer's disease. The distribution of galanthamine and four other alkaloids in different organs of Narcissus confusus plants, as well as the variations occurring during the ontogenic cycle of this plant species, was studied. The five alkaloids were found to be present in all the organs at every stage, with the exception of haemanthamine in senescent flowers. The highest amount of alkaloids occurred in the bulb at the emerging stage, galanthamine being the most abundant, reaching a concentration of up to 2.5 % referred to dry weight.
Assuntos
Alcaloides/biossíntese , Galantamina/biossíntese , Narcissus/metabolismo , Fitoterapia , Alcaloides/química , Galantamina/química , Humanos , Narcissus/crescimento & desenvolvimentoRESUMO
A novel, fast and precise method, combining solid-phase extraction and reversed-phase high-performance liquid chromatography is described for the quantitative determination of five alkaloids (galanthamine, N-formylnorgalanthamine, haemanthamine, homolycorine and tazettine/pretazettine) from bulbs of wild Narcissus confusus, a high galanthamine-containing plant species growing in the Iberian Peninsula.