COMPOSITION CONTAINING CAROTENOIDS AND USE THEREOF FOR PROTECTING NEURONS AGAINST NEURODEGENERATION

Abstract

Composition containing carotenoid pigments of the xanthophyll family, characterized in that said pigments of the xanthophyll family comprise at least diadinoxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of diadinoxanthin and/or violaxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of violaxanthin, for use in the prevention or treatment of cognitive decline or neurodegenerative diseases.

Claims

1-11. (canceled)

12. A medicament for use in the prevention or treatment of cognitive decline or neurodegenerative diseases containing carotenoid pigments of the xanthophyll family, characterized in that said pigments of the xanthophyll family comprise at least one of the molecules of the following formula (I) or (II): ##STR00003## ##STR00004## in which the R1, R2, R3 and R4 radicals, identical or different, are selected from hydrogen, a saturated or unsaturated, linear or branched hydrocarbon radical, such as a C.sub.1 to C.sub.36 alkyl radical, or the groups -OR1, -OR2, -OR3 or -OR4 are C.sub.4 to C.sub.36, preferably C.sub.4 to C.sub.28, fatty acid esters.

13. The medicament of claim 12, characterized in that the molecule of the xanthophyll family of formula (I) is diadinoxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of diadinoxanthin.

14. The medicament of claim 12, characterized in that the molecule of the xanthophyll family of formula (II) is violaxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of violaxanthin.

15. The medicament of claim 12, characterized in that the xanthophyll molecules are extracted from marine or freshwater microalgae, marine macroalgae, cyanobacteria or bacteria.

16. The medicament of claim 12, characterized in that the xanthophyll molecules of formula (I) are extracted from euglenophyte algae of the Euglenozoa phylum, diatoms of the Bacillariophyta phylum, dinoflagellates of the Dinophyceae class, Haptophytes, Pelagophytes, Phaetothamniophytes, Dictyochophytes or Ochrophytes.

17. The medicament of claim 12, characterized in that the xanthophyll molecules of formula (II) are extracted from algae of the phyla Chrysophytes, Eustigmatophytes, Synurophytes, Mesostigmatophytes, Chlorophytes, Prasinophytes, Chlorarachniophytes, Pinguiophytes, Raphidophytes, or extracted from the dinoflagellates, or from macro-algae.

18. A method for preparing the medicament of claim 12, comprising obtaining the xanthophyll molecules of formula (I) or (II) from microalgae by the following successive steps: i) grinding the microalgae and dispersion in ethanol in a ball mixer-grinder allowing the extraction of the xanthophylls, ii) after elimination of said balls, centrifugation for recovering the supernatant, iii) filtering the supernatant over a 0.2 .Math.m filter, iv) followed immediately by a separation and purification of the pigments by centrifugal partition chromatography or by high-performance liquid chromatography and UV detection of the eluted fractions, and v) drying and packaging.

19. A food supplement or a nutritional supplement comprising carotenoid pigments of the xanthophyll family, characterized in that said pigments of the xanthophyll family comprise at least one of the molecules of the following formula (I) or (II): ##STR00005## ##STR00006## in which the R1, R2, R3 and R4 radicals, identical or different, are selected from hydrogen, a saturated or unsaturated, linear or branched hydrocarbon radical, such as a C.sub.1 to C.sub.36 alkyl radical, or the groups -OR1, -OR2, -OR3 or -OR4 are C.sub.4 to C.sub.36, preferably C.sub.4 to C.sub.28, fatty acid esters.

20. The food supplement or a nutritional supplement of claim 19, characterized in that the molecule of the xanthophyll family of formula (I) is diadinoxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of diadinoxanthin.

21. The food supplement or a nutritional supplement of claim 19, characterized in that the molecule of the xanthophyll family of formula (II) is violaxanthin, or a C.sub.4 to C.sub.36 fatty acid ester of violaxanthin.

22. The food supplement or a nutritional supplement of claim 19, characterized in that the xanthophyll molecules are extracted from marine or freshwater microalgae, marine macroalgae, cyanobacteria or bacteria.

23. The food supplement or a nutritional supplement of claim 19, characterized in that the xanthophyll molecules of formula (I) are extracted from euglenophyte algae of the Euglenozoa phylum, diatoms of the Bacillariophyta phylum, dinoflagellates of the Dinophyceae class, Haptophytes, Pelagophytes, Phaetothamniophytes, Dictyochophytes or Ochrophytes.

24. The food supplement or a nutritional supplement of claim 19, characterized in that the xanthophyll molecules of formula (II) are extracted from algae of the phyla Chrysophytes, Eustigmatophytes, Synurophytes, Mesostigmatophytes, Chlorophytes, Prasinophytes, Chlorarachniophytes, Pinguiophytes, Raphidophytes, or extracted from the dinoflagellates, or from macro-algae.

25. A method for the prevention or the treatment of Alzheimer’s disease, Parkinson’s disease, epilepsy, learning disabilities/autism, spinocerebellar ataxia, multiple sclerosis, Lewy body dementia, Alexander disease, Alpers disease, multiple system atrophy, posterior cortical atrophy (Benson’s syndrome), cortico-basal degeneration, progressive supranuclear palsy, Pick’s disease, macrophagic myofasciitis, Huntington’s disease, amyotrophic lateral sclerosis, Creutzfeldt-Jakob disease, or glaucomatous neurodegeneration comprising administering the medicament of claim 12 to a human subject.

26. The food supplement or a nutritional supplement of claim 19, characterized in that it comprises at least one additional element selected from: the following vitamins: E, B3, B5, B6, H, B9 and B12; the following trace elements and minerals: magnesium, iodine and selenium; the following compounds: eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and sunflower lecithin; plant proteins; an extract of blueberry or another plant; the following carotenoids: astaxanthin, fucoxanthin, lutein, zeaxanthin, alpha-carotene, beta-carotene, crocetin, crocin, beta-cryptoxanthin and lycopene; excipients, emulsifiers, bulking agents, thickeners, anti-caking agents, stabilizers, acidifiers and flavourings.

27. The medicament of claim 12, characterized in that it comprises at least one additional element selected from: the following vitamins: E, B3, B5, B6, H, B9 and B12; the following trace elements and minerals: magnesium, iodine and selenium; the following compounds: eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and sunflower lecithin; plant proteins; an extract of blueberry or another plant; the following carotenoids: astaxanthin, fucoxanthin, lutein, zeaxanthin, alpha-carotene, beta-carotene, crocetin, crocin, beta-cryptoxanthin and lycopene; excipients, emulsifiers, bulking agents, thickeners, anti-caking agents, stabilizers, acidifiers and flavourings.

28. The medicament of claim 12, in the form of a hard capsule, a soft capsule, a liquid stick, a liquid, a tablet, a gel, a powder, a drinkable ampoules, an injectable form or a dropper form.

29. The food supplement or a nutritional supplement of claim 19, in the form of a hard capsule, a soft capsule, a liquid stick, a liquid, a tablet, a gel, a powder, a drinkable ampoules, an injectable form or a dropper form.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0047] The invention will be well understood on reading the following description of embodiments, with reference to the attached drawings in which:

[0048] FIG. 1 is a diagram showing the cell viability of HT22 neuronal cells treated either with 50 .Math.M of different xanthophyll molecules (white rectangles) for two hours followed by cell death induced by 10 mM glutamate, after incubation over 24 hours at 37° C. (black rectangles);

[0049] FIG. 2 is a graph showing cell viability in percentages as a function of the concentration in .Math.M of diadinoxanthin alone (dotted line) and diadinoxanthin in the presence of glutamate (solid line);

[0050] FIG. 3 is a graph showing cell viability in percentages as a function of the concentration in .Math.M of violaxanthin alone (dotted line) and violaxanthin in the presence of glutamate (solid line);

[0051] FIG. 4 is a comparative example showing cell viability in percentages as a function of the concentration in .Math.M of diatoxanthin alone (dotted line) and diatoxanthin in the presence of glutamate (solid line).

EXAMPLES

[0052] Example 1: pure molecules of diadinoxanthin and violaxanthin (marketed by SIGMA ALDRICH) as well as other xanthophyll molecules such as canthaxanthin, astaxanthin, diatoxanthin, fucoxanthin, fucoxanthinol, lutein and zeaxanthin were tested.

[0053] Violaxanthin can be extracted from Chlorella vulgaris or from Tetraselmis suecica. Diadinoxanthin can be extracted from the microalgae Odontella aurita or Isochrysis galbana.

[0054] The HT22 neuronal cells (mouse hippocampus neuronal line) were treated for two hours with 50 .Math.M of pigment, followed or not by the addition of 10 mM glutamate for 24 hours at 37° C. Cell viability (in %) was measured by an MTS test (by means of a CellTiter 96®Aqueous non-Radioactive Cell Proliferation Assay kit from Promega). The values shown in FIG. 1 are the mean ± standard deviation (n=3), .star-solid. P<0.05; .star-solid..star-solid. P<0.01; .star-solid..star-solid..star-solid. P<0.001 with respect to the untreated cell control. # P<0.05; ### P<0.001 with respect to the glutamate-induced cell control.

[0055] These results show the non-toxic effect at 50 .Math.M of most of the pigments tested (white rectangles) and the protective effect at over 50% of the diadinoxanthin and the violaxanthin against the cytotoxicity of glutamate on these HT22 neuronal cells (black rectangles). The morphology of the cells was obtained by INCUCYTE® S3.

[0056] It can thus be seen that the diadinoxanthin and violaxanthin pigments are non-toxic and generate a protective effect on cell death of the HT22 neuronal cells induced by glutamate at 10 mM (FIG. 1).

[0057] Example 2: following the interesting results given in Example 1 for diadinoxanthin and the violaxanthin, dose-response tests between 1 and 50 .Math.M on the same HT22 cell model induced by glutamate (10 mM) were conducted.

[0058] The HT22 cells were treated for 2 hours either with diadinoxanthin (FIG. 2) or violaxanthin (FIG. 3) or diatoxanthin (comparative test shown in FIG. 4) at different concentrations. For each of the concentrations, cell death was then induced by glutamate (10 mM) for 24 hours at 37° C. Cell viability (in %) was measured by an MTS test. The values shown are the mean ± standard deviation (n=3).

[0059] The EC50 (half maximal effective concentration) is the concentration at which a molecule induces a response halfway between the baseline and the maximum effect after a certain exposure time thereof. This determines its efficacy. For diadinoxanthin the EC50 value is 22 .Math.M and that of violaxanthin is 28 .Math.M.

[0060] These results clearly demonstrate that these two molecules protect neurons against cell death, this surprising effect being greater than that obtained by the other xanthophyll molecules already validated on animals for reducing the risk of neurodegenerative diseases (Cho et al., publication cited above).