New method for the study of Amaryllidaceae alkaloid biosynthesis using biotransformation of deuterium-labeled precursor in tissue cultures.

Tahchy, Anna El; Boisbrun, Michel; Ptak, Agata; Dupire, François; Chrétien, Françoise; Henry, Max; Chapleur, Yves; Laurain-Mattar, Dominique

doi:10.18388/abp.2010_2375

Cited by 29 publications

(38 citation statements)

References 27 publications

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“…These results showed that only at 0.1 g/L, the precursor enhanced the galanthamine accumulation in the shoot cultures. This latter alkaloid was either partially released into the liquid medium or partially transformed in other compounds as shown in a previous study [20], where labeled narwedine and N-formylgalanthamine were found in the shoot cultures. In control shoot cultures, lycorine was detected in the tissues (0.04 mg/g DW) at a constant level throughout the whole culturing period.…”

Section: Resultssupporting

confidence: 68%

4′‐O‐Methylnorbelladine feeding enhances galanthamine and lycorine production by Leucojum aestivum L. shoot cultures

Saliba

Ptak

Laurain-Mattar

2015

Engineering in Life Sciences

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Galanthamine and lycorine are Amaryllidaceae alkaloids that exhibit a wide range of pharmacological activities. Organic synthesis and plant extraction are the two main commercial sources of galanthamine. Since total organic synthesis is low yielding and complex, and the harvesting of plant biomass is causing wild plant depletion, in vitro cultures offer an alternative approach. Amaryllidaceae alkaloids can be regarded as derivatives of the common precursor 4′‐O‐methylnorbelladine via intramolecular oxidative phenol coupling. The aim of this work was to enhance the biosynthetic pathway of these alkaloids using in vitro techniques. The precursor 4′‐O‐methylnorbelladine was incorporated into the liquid medium of Leucojum aestivum shoot cultures at different concentrations (0, 0.05, 0.1, or 0.2 g/L). The cultures were conducted for various periods of time (15, 30, and 40 days). After alkaloid extraction and according to LC‐ESI‐MS analysis, the precursor highly stimulated the biosynthesis of both galanthamine (0.5 mg/g dry weight [DW], V/S 0.01 mg/g DW in the control sample, i.e. without precursor) and lycorine (0.2 mg/g DW, V/S 0.04 mg/g DW in the control sample) after its biotransformation by the shoot cultures. The optimal culture conditions for the production of both alkaloids were 0.1 g/L of precursor and 15 days of culture.

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Section: Resultssupporting

confidence: 68%

4′‐O‐Methylnorbelladine feeding enhances galanthamine and lycorine production by Leucojum aestivum L. shoot cultures

Saliba

Ptak

Laurain-Mattar

2015

Engineering in Life Sciences

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“…Control medium free of MN was also used. The cultures were in-cubated for various periods of time (15,30,40 and 50 days) under white fluorescent light with a 16 h photoperiod (Tungsram lamp, 40 WF, 90 μmol m −2 s −1 ) at 25 ± 2°C. Three replicates for each condition were made.…”

Section: Bulblet Culturesmentioning

confidence: 99%

“…In a previous work [30], deuterated MN was prepared according to a method reported by Szewczyk et al [42] to synthesize the undeuterated precursor. It involved a reductive amination of isovanillin with tyramine.…”

Section: Optimization Of 4'-o-methylnorbelladine (Mn) Synthesismentioning

confidence: 99%

Stimulating effect of both 4’‐O‐methylnorbelladine feeding and temporary immersion conditions on galanthamine and lycorine production by Leucojum aestivum L. bulblets

Saliba

Ptak

Boisbrun

et al. 2016

Engineering in Life Sciences

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Bulb cultures of Leucojum aestivum and L. aestivum 'Gravety Giant' were subcultured in medium containing the precursor 4'-O-methylnorbelladine (MN) at various concentrations [0 (control), 0.15 and 0.3 g/L]. The cultures were conducted in bioreactor RITA R and lasted for 15, 30, 40 and 50 days. The growth rate and the alkaloid accumulation in bulblets were studied. For this latter purpose, a purification method was developed. It comprised a highly selective solid phase extraction using on the one hand, UPTI-CLEAN SI and SCX cartridges for plant extracts and on the other hand, 2H cartridges for culture media. Pure alkaloidal fractions were, thus, analyzed by LC-ESI-MS allowing the quantitative evaluation of galanthamine and lycorine from culture extracts. Precursor feeding along with temporary immersion conditions was found to significantly improve the accumulation of both galanthamine and lycorine. The maximal concentrations of galanthamine (0.81 mg/g DW) and lycorine (0.54 mg/g DW) in L. aestivum bulblets were reached, respectively, after 40 days of culture with 0.15 g/L of precursor and after 30 days of culture with 0.3 g/L of precursor. In L. aestivum 'Gravety Giant' bulb cultures, 0.3 g/L of precursor was the best condition for both galanthamine (0.6 mg/g DW after 50 days) and lycorine (1.13 mg/g DW after 30 days).

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“…24,[30][31][32]36 In vitro studies in such settings have shown that lycorine can induce apoptosis, specifically targeted against malignant cells. 31 Its potential use as a therapeutic agent has recently led to studies into the production of the synthetic compound, 40,41 highlighting it as a potential lead for drug development. 24,42 In this study we investigated the potential of combining lycorine with bezafibrate + MPA to elicit an apoptotic response in the presence of CD40L and assessed the correlation between induced apoptosis and the generation of mitochondrial superoxide.…”

Section: Introductionmentioning

confidence: 99%

Lycorine sensitizes CD40 ligand-protected chronic lymphocytic leukemia cells to bezafibrate- and medroxyprogesterone acetate-induced apoptosis but dasatanib does not overcome reported CD40-mediated drug resistance

Hayden¹,

Pratt²,

Drayson³

et al. 2010

Haematologica

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BackgroundTumor cells in chronic lymphocytic leukemia accumulate in the periphery through the proliferation of a minority of cells in lymph nodes. The proliferative and survival signals in these proliferation centers include interactions with T lymphocytes expressing CD40 ligand. We have demonstrated that the low toxicity combination of bezafibrate and medroxyprogesterone acetate induces mitochondrial superoxide-mediated apoptosis of non-CD40-liganded cells but not of cells exposed to CD40 ligand. Here, we assessed the ability of dasatinib and lycorine to restore bezafibrate-and medroxyprogesterone acetate-induced apoptosis in cells exposed to CD40 ligand. In parallel experiments we compared the ability of dasatinib to induce apoptosis of cells co-treated with fludarabine. Design and MethodsPrimary chronic lymphocytic leukemia and peripheral blood mononuclear cells were exposed to drug combinations for 72 hours on control and CD40 ligand-expressing fibroblast monolayers. Cells were harvested and analyzed for apoptosis and levels of mitochondrial superoxide using flow cytometry. In some experiments cells were removed from CD40 ligand at 48 hours, retreated and analyzed after a further 24 hours. The effect of CD40 ligand and drug treatments on mitochondrial superoxide levels were assessed. ResultsAs previously described, dasatinib rendered cells sensitive to fludarabine but only when CD40 ligand was removed for the last 24 hours of culture. In contrast, lycorine restored the bezafibrate-and medroxyprogesterone acetate-induced apoptosis associated with mitochondrial superoxide even during continuous exposure to CD40 ligand. Furthermore, combined bezafibrate, medroxyprogesterone acetate and lycorine had little effect against normal peripheral blood mononuclear cells, whereas dasatinib with fludarabine induced high levels of apoptosis. ConclusionsOur data indicate the potential of bezafibrate, medroxyprogesterone acetate and lycorine as novel therapy in chronic lymphocytic leukemia and have important implications for the reported potential of c-abl kinase inhibitors in this disease.Key words: bezafibrate, medroxyprogesterone acetate, lycorine, chronic lymphocytic leukemia, mitochondrial superoxide, CD40 ligand. CD40-mediated drug resistance. Haematologica 2010;95(11):1889-1896. doi:10.3324/haematol.2010 This is an open-access paper. Citation: Hayden RE, Pratt G, Drayson MT, and Bunce CM. Lycorine sensitizes CD40 ligand-protected chronic lymphocytic leukemia cells to bezafibrate-and medroxyprogesterone acetate-induced apoptosis but dasatanib does not overcome reportedLycorine sensitizes CD40 ligand-protected chronic lymphocytic leukemia cells to bezafibrate-and medroxyprogesterone acetate-induced apoptosis but dasatanib does not overcome reported CD40-mediated drug resistance

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New method for the study of Amaryllidaceae alkaloid biosynthesis using biotransformation of deuterium-labeled precursor in tissue cultures.

Cited by 29 publications

References 27 publications

4′‐O‐Methylnorbelladine feeding enhances galanthamine and lycorine production by Leucojum aestivum L. shoot cultures

4′‐O‐Methylnorbelladine feeding enhances galanthamine and lycorine production by Leucojum aestivum L. shoot cultures

Stimulating effect of both 4’‐O‐methylnorbelladine feeding and temporary immersion conditions on galanthamine and lycorine production by Leucojum aestivum L. bulblets

Lycorine sensitizes CD40 ligand-protected chronic lymphocytic leukemia cells to bezafibrate- and medroxyprogesterone acetate-induced apoptosis but dasatanib does not overcome reported CD40-mediated drug resistance

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