María Lenny Rodríguez-Escobar

Amaryllidaceae alkaloids are pharmaceutically relevant specialized metabolites whose variable accumulation in planta has driven the development of controlled biotechnological production systems. This study developed an integrated Hippeastrum × hybridum cell-culture platform encompassing explant selection, callus induction, suspension culture establishment, bioreactor cultivation, and elicitation. Different explants were evaluated for callogenic and morphogenic competence, selected callus lines were adapted to suspension culture. B1- and BS-derived cultures were progressively expanded and transferred to 2 L bioreactors as an intermediate scale-up step and subsequently exposed to 1 µM coronatine (COR). Alkaloid extracts from cellular and extracellular fractions, together with in vitro and ex vitro plant tissues, were analyzed by GC–MS. Explant origin strongly influenced developmental responses, and B1- and BS-derived lines were selected based on sustained callus proliferation, biomass availability, suitable friability, and successful adaptation to liquid culture. Both lines maintained growth after transfer to 2 L bioreactors; in the bioreactor cultures evaluated, B1 showed greater estimated net biomass accumulation than BS. Their alkaloid-related profiles differed markedly between lines and sampling times during the elicitation phase. Notably, an extracellular signal assigned to O-methylnorbelladine was detected in B1 at D7, consistent with retention of metabolic competence related to Amaryllidaceae alkaloid biosynthesis. The developed system should therefore be regarded as a proof-of-concept laboratory-scale platform rather than an optimized alkaloid-production process, providing a basis for future studies with biological replication, quantitative process optimization, and application to Hippeastrum species or genotypes preselected for pharmaceutically relevant alkaloid profiles.

The Amaryllidaceae family is a rich source of structurally diverse alkaloids with recognized neuroactive and antiparasitic properties. This study provides the first phytochemical and biological characterization of Pyrolirion boliviense and wild Stenomesson miniatum from Bolivia. Alkaloid extracts from bulbs and leaves were analysed by GC–MS and evaluated for acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and Trypanosoma cruzi inhibitory activities. Thirty-two Amaryllidaceae alkaloids were identified, with P. boliviense exhibiting greater alkaloid diversity (25 compounds) and S. miniatum a higher total alkaloid content (227.86 vs. 138.92 μg Gal/100 mg DW). P. boliviense bulb extracts showed the strongest cholinesterase inhibition (AChE IC₅₀ = 6.07 ± 0.47 μg·mL⁻¹; BuChE IC₅₀ = 30.93 ± 1.17 μg·mL⁻¹), whereas S. miniatum extracts displayed weaker AChE inhibition and no detectable BuChE activity. In anti-T. cruzi assays, bulb extracts were the most active, with S. miniatum showing an IC₅₀ of 0.90 ± 0.15 μg·mL⁻¹ (SI = 20.12) and selective anti-amastigote activity (IC₅₀ = 1.42 ± 0.66 μg·mL⁻¹; SI = 12.77). These findings identify Bolivian Andean Amaryllidaceae as promising sources of bioactive alkaloids with potential applications for Alzheimer’s disease and Chagas disease drug discovery.

Molecular diversity is a key component of overall biodiversity, playing a vital role in evolution. It results from the adaptation of organisms to various habitats, which impacts their survival. The Amaryllidoideae subfamily is a significant group of monocotyledonous plants known for producing an exclusive and still-expanding group of molecules with diverse biological activities. Galanthamine (Gal), the most renowned metabolite from Amaryllidoideae subfamily, has been marketed for the palliative treatment of Alzheimer’s disease since 2001 due to its ability to inhibit the acetylcholinesterase enzyme. Due to the high cost and low yield of its synthesis, pharmaceutical companies extract this drug from Amaryllidoideae plants, such as Narcissus pseudonarcissus cv. Carlton in Europe and Lycoris radiata in China. The aim of this study was to describe the alkaloid profile of fifteen different species of Narcissus L. (commonly known as daffodils) collected in Spain using gas chromatography coupled with mass spectrometry. Fifty-one alkaloids were identified and quantified within these species through our private library of Amaryllidaceae alkaloids (AA) built over the last four decades, while thirty structures remained not identified in thirteen of these species. The highest concentration of these nitrogenate metabolites was quantified in N. confusus, 541 μg Gal·100 mg⁻¹ DW, which also exhibited a notably high concentration of Gal, 301 μg Gal·100 mg⁻¹ DW, which represents about 55% of the alkaloids identified in this species. The species N. bujei was also found to contain a significant quantity of this compound, amounting to 103.2 μg Gal·100 mg⁻¹ DW. The plant N. assoanus harbored a total of seven unidentified compounds, indicating that this species could be a potentially important source of novel alkaloids. In conclusion, this study facilitates a direct comparison of alkaloid profiles for fifteen Narcissus plant species. This serves as a valuable tool for identifying possible new sources of galanthamine, as well as other novel medicinal alkaloids. Finally, this work presents the first alkaloid profile of the species N. minor and N. nevadensis.

Amaryllidaceae alkaloids from the Amaryllidoideae subfamily exhibit broad pharmacological activities, including neuroprotection and anticancer effects. Galanthamine is a key compound for Alzheimer’s therapy. The Hippeastrum genus, particularly in Bolivia, offers significant potential for novel drug discovery, emphasising the need for conservation and further phytochemical research. Twenty-seven samples from Bolivian Hippeastrum species were investigated in terms of their alkaloid profile and anticholinesterase activity. The phytochemical analysis of Bolivian Hippeastrum species via GC-MS identified 48 Amaryllidaceae alkaloids, displaying diverse structural groups with potential pharmacological significance. Lycorine- and Homolycorine-type alkaloids were predominant, particularly in H. chionedyanthum and H. haywardii, with high concentrations of lycorine, a promising anticancer compound. The species H. evansiarum and H. mollevillquense contained notable quantities of Galanthamine type alkaloids, relevant for Alzheimer’s treatment. This study also highlights variability in acetylcholinesterase and butyrylcholinesterase inhibitory activities, with H. lara-ricoi and H. haywardii demonstrating strong inhibition. These findings suggest that Hippeastrum species are a valuable source of bioactive compounds, warranting further research into their therapeutic applications.

Narcissus L. is a renowned plant genus with a notable center of diversity and is primarily located in the Mediterranean region. These plants are widely recognized for their ornamental value, owing to the beauty of their flowers; nonetheless, they also hold pharmacological importance. In Europe, pharmaceutical companies usually use the bulbs of Narcissus pseudonarcissus cv. Carlton to extract galanthamine, which is one of the few medications approved by the FDA for the palliative treatment of mild-to-moderate symptoms of Alzheimer’s disease. The purpose of this study was to evaluate the potential of these plants in Alzheimer’s disease. The alkaloid extract from the leaves of different species of Narcissus was obtained by an acid-base extraction work-up -procedure. The biological potential of the samples was carried out by evaluating their ability to inhibit the enzymes acetyl- and butyrylcholinesterase (AChE and BuChE, respectively). The species N. jacetanus exhibited the best inhibition values against AChE, with IC₅₀ values of 0.75 ± 0.03 µg·mL⁻¹, while N. jonquilla was the most active against BuChE, with IC₅₀ values of 11.72 ± 1.15 µg·mL⁻¹.

The genus Clinanthus Herb. is found in the Andes Region (South America), mainly in Peru, Ecuador, and Bolivia. These plants belong to the Amaryllidaceae family, specifically the Amaryllidoideae subfamily, which presents an exclusive group of alkaloids known as Amaryllidaceae alkaloids that show important structural diversity and pharmacological properties. It is possible to find some publications in the literature regarding the botanical aspects of Clinanthus species, although there is little information available about their chemical and biological activities. The aim of this work was to obtain the alkaloid profile and the anti-cholinesterase activity of four different samples of Clinanthus collected in South America: Clinanthus sp., Clinanthus incarnatus, and Clinanthus variegatus. The alkaloid extract of each sample was analyzed by gas chromatography coupled with mass spectrometry (GC-MS), and their potential against the enzymes acetyl- and butyrylcholinesterase were evaluated. Thirteen alkaloids have been identified among these species, while six unidentified structures have also been detected in these plants. The alkaloid extract of the C. variegatus samples showed the highest structural diversity as well as the best activity against AChE, which was likely due to the presence of the alkaloid sanguinine. The results suggest this genus as a possible interesting new source of Amaryllidaceae alkaloids, which could contribute to the development of new medicines.

Amaryllidaceae alkaloids are secondary metabolites with interesting medicinal properties. Almost every Narcissus species can synthesize them and constitute an excellent source for their isolation and study. Several Amaryllidaceae alkaloids have shown acetylcholinesterase inhibitory activities and are a promising tool for treating cholinergic disorders such as Alzheimer’s disease (AD). Indeed, three of the four palliative treatments approved for AD are acetylcholinesterase (AChE) inhibitors and one of them, galanthamine, is an Amaryllidaceae alkaloid itself. This molecule is currently isolated from natural sources. However, its production is insufficient to supply the increasing demand for the active principle. Our main aim is to discover tools to improve galanthamine production and to prospect for potential new and more efficient drugs for AD treatment. Furthermore, we seek to broaden the knowledge of plants of the genus Narcissus from a chemotaxonomic perspective. Hence, in this study, we evaluate the alkaloid content through GC–MS and the AChE inhibitory activity of ten autumn-flowering Narcissus, which have been less studied than their spring-flowering counterparts. A total of thirty Amaryllidaceae alkaloids have been found, twenty-eight properly identified. Two Narcissus contained galanthamine, and seven were able to inhibit AChE.

Plant biodiversity is an important source of compounds with medicinal properties. The alkaloid galanthamine, first isolated from Galanthus woronowii (Amaryllidaceae), is approved by the FDA for the palliative treatment of mild to moderate Alzheimer’s disease due to its acetylcholinesterase (AChE) inhibitory activity. Obtaining this active pharmaceutical ingredient, still sourced on an industrial scale from the Amaryllidaceae species, is a challenge for pharmaceutical companies due to its low natural yield and the high cost of its synthesis. The aim of this work was to determine the alkaloid profile of three different Rauhia (Amaryllidaceae) species collected in Peru, and to assess the potential application of their extracts for the treatment of Alzheimer’s disease. The alkaloids were identified by gas chromatography coupled to mass spectrometry (GC-MS), and the AChE inhibitory activity of the extracts was analyzed. Thirty compounds were quantified from the Rauhia species, the R. multiflora extract being the most interesting due to its high diversity of galanthamine-type structures. The R. multiflora extract was also the most active against AChE, with the half maximal inhibitory concentration (IC₅₀) values of 0.17 ± 0.02 μg·mL⁻¹ in comparison with the IC₅₀ values of 0.53 ± 0.12 μg·mL⁻¹ for galanthamine, used as a reference. Computational experiments were carried out on the activity of the galanthamine-type alkaloids identified in R. multiflora toward five different human AChE structures. The simulation of the molecules 3-O-acetylgalanthamine, 3-O-acetylsanguinine, narwedine, and lycoraminone on the 4EY6 crystal structure theoretically showed a higher inhibition of hAChE and different interactions with the active site compared to galanthamine. In conclusion, the results of this first alkaloid profiling of the Rauhia species indicate that R. multiflora is an important natural source of galanthamine-type structures and could be used as a model for the development of biotechnological tools necessary to advance the sustainable production of galanthamine.