Composition to improve cognitive functions

Abstract

The invention relates to a composition comprising a mixture of catechin and / or epicatechin, safranal and crocetin in aglycone and / or glycosylated form. Such a composition can be used as a nutritional product or medicament and has a synergistic action for improving cognitive functions and / or executive functions in humans or animals.

Description

Title of the invention: Composition for improving cognitive functions

[0001] The invention relates to a composition comprising a mixture of molecules, in particular a mixture of catechin and / or epicatechin, safranal and crocetin (aglycone and / or glycosylated). Such a composition can be used as a nutritional product or medicament and has a synergistic action for improving cognitive functions and / or executive functions in humans or animals.

[0002] It is known that cognitive disorders and dementias, in particular neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease, mainly affect elderly people. Due to the evolution of medicine and the quality of care provided, the elderly population is increasing. There is therefore a high prevalence of these age-related disorders. It is therefore of interest to develop solutions to prevent or minimize age-related cognitive decline and delay the onset of associated pathologies, in particular to prolong the autonomy of the elderly population and thus maintain their quality of life.

[0003] To prevent or delay the progression to dementia and thus maintain a stable cognitive state and well-being in older people, it has been suggested to maintain a healthy diet. Nutrition has been suggested as an important factor in preventing the impairment of cognitive functions.

[0004] To this end, nutritional agents and food supplements have been developed. In particular, patent application FR 1560263 describes a mixture of molecules obtained from Vitis vinifera and Vaccinium angustifolium, comprising: at least 1% catechin and / or epicatechin, and at least 5 ppm ferulic acid. Such a mixture is therefore known as well as its effects on the cognitive functions of humans when taken chronically.

[0005] It is also known that specific diets can prevent or minimize age-related cognitive decline and thus delay the onset of associated pathologies. Thus, the Mediterranean diet and its derivatives such as the MIND or DASH diet appear to be beneficial in preventing cognitive decline during aging. This type of diet is rich in neuroprotective nutrients such as flavonoids and other polyphenols, carotenoids, omega 3 fatty acids, certain vitamins (folates, vitamin E in particular) and certain minerals have been the subject of numerous studies in order to identify their effects individually.

[0006] Fruits and vegetables rich in carotenoids, in particular lutein, zeaxanthin and their derivatives (aglycone and / or glycosylated crocetins), are also known to their effects on brain functions. Crocetin is an aglycone form of crocin that is metabolized into crocetin by the body, either after administration.

[0007] Also, the objective of the invention is to provide a composition having greater effectiveness in combating cognitive decline than specific molecules consumed individually or known mixtures of molecules.

[0008] To this end, the invention relates to a composition comprising a mixture of molecules comprising: - at least 0.15% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, - at least 15 ppm (parts per million in the mixture) of safranal by weight relative to the total weight of the mixture measured by HPLC method, and - at least 80 ppm of crocetin in aglycone and / or glycosylated form.

[0009] Thus, the composition according to the invention comprises flavanols, in particular catechins and / or epicatechins, combined with specific saffron molecules: safranal and crocetin in aglycone and / or glycosylated form.

[0010] Catechins and epicatechin are compounds of the flavonoid family, particularly of the flavanol subclass. They are also known as catechol and epicatechin and are powerful antioxidants that help prevent inflammatory and coronary diseases, but also maintain or increase cognitive performance.

[0011] Safranal is a volatile organic compound isolated from saffron and is notably responsible for its aroma. It can be extracted from several plants such as Crocus sativus, Centaurea sibthorpii, Centaurea consanguinea, Centaurea amanicola, Erodium cicutarium, Chinese green tea, Calycopteris floribunda, Crocus heuffelianus, Sambucus nigra, Gardenia jasminoides, Citrus limon, Cuminum cyminum L., Achillea distans but is mainly extracted from Crocus sativus.

[0012] Crocetins in aglycone and / or glycosylated form have a multifunctional role in neuroprotection, notably by modulating the aggregation of b-amyloid and tau peptides, slowing down cognitive decline and amnesic disorders (Finley JW, Gao S. A Perspective on Crocus sativus L. (Saffron) Constituent Crocin: A Potent Water-Soluble Antioxidant and Potential Therapy for Alzheimer's Disease. J Agric Food Chem. 2017 Feb 8;65(5): 1005-20). As previously, it can be extracted from Crocus sativus or Gardenia jasminoides and is notably mainly responsible for the color of saffron.

[0013] In the prior art, effects on cognitive decline have been suggested of these molecules individually, and of extracts comprising certain particular mixtures of flavanols on the one hand, and of aglycone and / or glycosylated safranal / crocetins on the other hand, but such a composition has not been described.

[0014] However, surprisingly, the specific combination as well as the quantity of molecules present in the present composition exhibit a synergistic effect and consequently, an improved effect on cognitive decline. The inventors have thus observed in tests on several markers of cognitive decline said synergistic effect in comparison with said molecules administered in isolation or via the extracts of existing plants known from the prior art or during the consumption of grapes, blueberries or saffron.

[0015] The invention also relates to a composition further comprising ferulic acid. Ferulic acid is known for its effects on the neuronal system, in particular to protect neuronal cells from cell death induced by cerebral ischemia (Cheng SY, et al. Ferulic acid provides neuroprotection against oxidative stress-related apoptosis after cerebral ischemia / reperfusion injury by inhibiting ICAM-1 mRNA expression in rats. Brain Res. 2008;1209:136-150). Furthermore, its antioxidant activity has been tested in Alzheimer's disease (Sgarbossa A, et al. Ferulic Acid: A Hope for Alzheimer's Disease Therapy from Plants. Nutrients. 2015;7(7):5764-82.) and its long-term administration appears to protect against memory and learning deficits (Yan JY et al. Protection against [3-amyloid peptide toxicity in vivo with long-term administration of ferulic acid. Br J Pharmacol. 2001;133:89-96.). Such a composition presents a further enhanced effect.

[0016] The effect of the composition is particularly increased and effective when it also comprises: - at least 30 ppm of resveratrol, and / or - at least 5 ppm of quercetin and / or quercetin glycosides, and / or - at least 50 ppm of anthocyanidins in aglycone and glycosylated forms.

[0017] Resveratrol is particularly known for being present in grapes. However, it can also be obtained from other plant sources, in particular from Polygonum cuspidatum but also by biotechnological means. It has many beneficial activities for humans or animals, including an improvement in working memory, learning, spatial memory and spontaneous motor activity (Abraham J, Johnson RW. Consuming a diet supplemented with resveratrol reduced infection-related neuroinflammation and deficits in working memory in aged mice. Rejuvenation Res [Internet]. 2010 / 01 / 01. 2009; 12(6):445-53).

[0018] Quercetin also has significant neuroprotective activity (Dajas F, et al. Neuroprotective actions of flavones and flavonols: mechanisms and relationship to flavonoid structural features. Cent Nerv Syst Agents Med Chem. 2013; 13(1):30-5) and it is also known that the consumption of foods rich in anthocyanidins (aglycones and / or glycosylated) helps prevent memory deficiencies and to improve cognitive performance (Barros D, et al. Behavioral and geno-protective effects of Vaccinium berries intake in mice. Pharmacol Biochem Behav.2006;84(2):229-34).

[0019] According to a preferred embodiment, the molecules present in the composition according to the invention are derived from a mixture of at least one extract of Vitis vinifera, at least one extract of Vaccinium angustifolium and at least one extract of Crocus sativus. Thus, the invention also relates to a composition comprising a mixture of at least one extract of Vitis vinifera, at least one extract of Vaccinium angustifolium and at least one extract of Crocus sativus.

[0020] The invention also relates to a composition for its use as a medicament or nutritional product, in particular in the prevention, alleviation or treatment of cognitive disorders.

[0021] Finally, the invention also relates to the non-therapeutic use of said composition in humans or animals.

[0022] The invention is now described in detail and with reference to the figures. Figures

[0023] [fig.lA] [fig.lA] represents the effect of the composition according to the invention and of the composition according to the prior art on the activity of AChE. Panel A represents the normalized absorbances (mean + SEM) over time and as a function of the different compositions tested.

[0024] [fig.lB] [fig.lB] represents the effect of the composition according to the invention and of the composition according to the prior art on the activity of AChE. Panel B represents the normalized absorbances (mean + SEM) at 40 min as a function of the different compositions tested as well as their respective percentages of inhibition compared to the control, n: number of replicates. P-value for the mixture of polyphenols: <0.0001, P-value for the mixture of carotenoids: <0.0001, P-value for the interaction: <0.0001.

[0025] [fig.2] [fig.2] represents the effect of the composition according to the invention and of com position according to the prior art on the survival of cortical neurons following the addition of glutamate in cell culture. The results (mean + SEM) are expressed as percentage survival compared to the control condition, n: number of replicates *** p<0.05, ** p<0.01 versus glutamate.

[0026] Definitions

[0027] By "at least X% of catechins and / or epicatechins" is meant either at least X% of catechins if there are no epicatechins in the mixture, or at least X% of epicatechins if there are no catechins in the mixture, or at least X% of the mixture of catechins and epicatechins if both catechins and epicatechins are present in the mixture. Preferably, at least X% of the mixture of catechins and epicatechins is meant.

[0028] By “ppm” we mean parts per million in the mixture.

[0029] For the purposes of the invention, the term "nutritional product" means a food ingredient for nutritional purposes used alone or combined with other ingredients or food additives in food formulas, including food supplements and foods intended for humans or animals.

[0030] For the purposes of the invention, the term "medicine" means an active ingredient used for therapeutic purposes, used alone or in combination with other active or non-active substances in medicinal formulas, including herbal medicine, intended for humans or animals. Detailed description

[0031] Composition

[0032] The subject of the invention is therefore a composition comprising a mixture of molecules comprising: - at least 0.15% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, - at least 15 ppm (parts per million in the mixture) of safranal by weight relative to the total weight of the mixture measured by HPLC method, and - at least 80 ppm of crocetin in aglycone and / or glycosylated form.

[0033] According to a preferred object of the invention, the composition comprises at least 0.9% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, preferably at least 15%.

[0034] Preferably, the composition comprises at least 50 ppm of safranal by weight relative to the total weight of the mixture measured by HPLC method, even more preferably at least 90 ppm

[0035] Preferably, the composition comprises at least 200 ppm of crocetin in aglycone and / or glycosylated form, even more preferably at least 1000 ppm of crocetin in aglycone or glycosylated form.

[0036] According to another embodiment, the composition and in particular the mixture further comprises at least 0.5 ppm of ferulic acid, preferably at least 4 ppm.

[0037] According to another particularly preferred embodiment of the invention, the composition also comprises: - at least 30 ppm of resveratrol, and / or - at least 5 ppm of quercetin and / or quercetin glycosides, and / or - at least 50 ppm of anthocyanidins.

[0038] According to a particularly preferred embodiment, the subject of the invention is a composition comprising a mixture of molecules containing: - at least 0.15% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, - at least 15 ppm (parts per million in the mixture) of safranal by weight relative to the total weight of the mixture measured by HPLC method, - at least 80 ppm of crocetin in aglycone or glycosylated forms, - at least 30 ppm of resveratrol, - at least 5 ppm of quercetin and / or quercetin glycosides, and - at least 50 ppm of anthocyanidins (in aglycone and / or glycosylated forms).

[0039] The composition or mixture of molecules according to the preceding embodiment may also contain possible residues resulting from at least one of the stages of preparation, obtaining or manufacturing of said composition.

[0040] For the purposes of the invention, the term "residues" means a residue or residues which may be any natural or synthetic molecule and which may be present in trace amounts and which have no effectiveness in the composition according to the present invention. The concept of residues may, for example, be residues of water (for example, less than 10% by weight of the total weight of the composition), alcohol (for example, less than 5% by weight of the total weight of the composition), enzyme, etc. For the purposes of the invention, the term "trace" means a product present in a very small quantity, in particular when the content makes the compound, molecule or product difficult to detect.

[0041] According to another particular embodiment, the composition according to the invention may consist exclusively of the mixture of molecules described according to any one of the preceding embodiments, or comprise other constituents. Preferably, said other constituents may be, in particular excipients, coating agents, such as maltodextrin, microcrystalline cellulose, cyclodextrins, starch, soluble or insoluble fibers, water or alcohol.

[0042] According to one embodiment, the molecules present in the composition are derived from Vitis vinifera and / or Vaccinium angustifolium and Crocus sativus.

[0043] Thus, the composition according to the invention may comprise a mixture of molecules derived from: - an extract of Vitis vinifera and / or an extract of Vaccinium angustifolium and an extract of Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera and Vaccinium angustifolium and an extract of Crocus sativus.

[0044] Preferably, the composition according to the invention comprises molecules which are derived from a mixture of at least one extract of Vitis vinifera, at least one extract of Vaccinium angustifolium and at least one extract of Crocus sativus.

[0045] According to one embodiment, the invention therefore relates to a composition comprising at least: - an extract of Vitis vinifera and / or an extract of Vaccinium angustifolium and an extract of Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera and Vaccinium angustifolium and an extract of Crocus sativus.

[0046] For the purposes of the invention, the term “Vitis vinifera extract” means at least one molecule, preferably a set of molecules, obtained from Vitis vinifera. The raw material may be the leaves and / or the fruits and / or the seeds and / or the skins and the wood, preferably the raw material is the aerial part of the plant, i.e. all the leaves, fruits, skin, seeds and wood, even more preferably the skin and the seeds.

[0047] For the purposes of the invention, the term “extract of Vaccinium angustifolium” means at least one molecule, preferably a set of molecules, obtained from Vaccinium angustifolium. The raw material may be the leaves and / or the fruits, preferably the raw material is all the leaves and fruits of the plant.

[0048] Preferably, the mixture of Vitis vinifera and Vaccinium angustifolium comprises or contains at least 1% of catechin and / or epicatechin of the total weight of said extract, preferably at least 5%.

[0049] According to another preferred embodiment, the mixture of Vitis vinifera and Vaccinium angustifolium further comprises or contains at least 5 ppm of ferulic acid, at least 200 ppm of resveratrol, 50 ppm of quercetins and / or quercetin glycosides and / or at least 500 ppm of anthocyanins (glycosylated and aglycones). Thus, the mixture of Vitis vinifera and Vaccinium angustifolium may comprise or contain at least 1% of catechin and / or epicatechin of the total weight of said extract, at least 5 ppm of ferulic acid, at least 200 ppm of resveratrol, 50 ppm of quercetins and / or quercetin glycosides and / or at least 500 ppm of anthocyanins.

[0050] For the purposes of the invention, the term “Crocus sativus extract” means at least one molecule, preferably a set of molecules, obtained from Crocus sativus. The raw material may be the stigmas and / or the petals and / or the bulbs of the plant, preferably the raw material is all the stigmas, petals and bulbs of the plant. Preferably, the Crocus sativus extract is an extract having a safranal content of at least 0.2% by weight of the total weight of the extract. of saffron and / or at least 1% of crocetin in aglycone and / or glycosylated form, 0.5% of picrocrocin and 500 ppm of flavonoids derived from kaempferol of the total weight of said extract.

[0051] By extract obtained from a mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus is meant a set of molecules obtained from a mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus.

[0052] Preferably the mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus contains: - at least 0.15% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, preferably at least 0.9%, even more preferably at least 15%. - at least 15 ppm (parts per million in the mixture) of safranal by weight relative to the total weight of the mixture measured by HPLC method, preferably at least 50 ppm, even more preferably at least 90 ppm and - at least 80 ppm of crocetin in aglycone and / or glycosylated form, preferably at least 200 ppm, even more preferably at least 1000 ppm.

[0053] Even more preferably, the mixture further comprises: - at least 0.5 ppm of ferulic acid, preferably at least 4 ppm and / or - at least 30 ppm of resveratrol, and / or - at least 5 ppm of quercetin and / or quercetin glycosides, and / or - at least 50 ppm of anthocyanins in aglycone and / or glycosylated form.

[0054] The raw material of Vitis vinifera may be the leaves and / or the fruits and / or the seeds and / or the wood, preferably the raw material of Vitis vinifera is the aerial part of the plant, that is to say all the leaves, fruits, skin, seeds and wood. The raw material of Vaccinium angustifolium may be the leaves and / or the fruits, preferably the raw material of Vaccinium angustifolium is all the leaves and fruits of the plant. The raw material of Crocus sativus may be the stigmas and / or the petals and / or the bulbs, preferably the raw material of Crocus sativus is all the stigmas, petals and bulbs of the plant.

[0055] The composition according to the invention may comprise, in addition to the mixture of molecules described above, at least one other constituent chosen from antioxidants, natural extracts, vitamins, trace elements, carotenoids, omega 3 fatty acids, natural oils, and mixtures thereof.

[0056] Examples of such antioxidant agents include polyphenols, in particular in the form of plant extracts (extracts of olive leaves or olives, blackcurrant, coffee, polygonum, citrus fruits, capers), and vitamin C, in particular in the form of plant extracts (acerola extract, pomegranate extract, citrus fruits), vitamin E, vitamin A, in particular in the form of plant extracts; or their derivatives, zinc, selenium and their mixtures.

[0057] As examples of plant extracts, in addition to those of said invention, mention may be made of extracts of coffee, olive leaves, blackcurrant, polygonum, citrus fruits, marigold and their mixtures.

[0058] Examples of vitamins include vitamin B9, B12, C, D, E, A including pro-vitamins A, their derivatives and their mixtures.

[0059] Vitamin D plays a key role in the central nervous system. In humans, several longitudinal studies have reported that vitamin D deficiency or deficiency is associated with an increased risk of developing Alzheimer's disease in later years. However, only one intervention study has shown the effect of vitamin D supplementation on memory and learning tests in older women.

[0060] B vitamins, particularly vitamins B9 and B12, also play an important role in brain function. Epidemiological studies have shown associations between B vitamin intake, or serum markers of these vitamins, and a lower risk of dementia or cognitive decline. In addition, higher dietary intakes of folate (vitamin B9) have been shown in humans to be associated with a significant reduction in the risk of dementia and Alzheimer's disease after several years of follow-up. In addition, a review evaluating the value of different types of intervention aimed at preventing cognitive decline and Alzheimer's disease recently showed that folic acid and vitamin B12 supplementation could have benefits on memory.

[0061] Examples of trace elements include zinc, selenium or chromium and mixtures thereof.

[0062] Zinc is an essential mineral for proper brain function by improving insulin sensitivity, reducing oxidative stress and inflammation. Moreover, the European Food Safety Agency, within the framework of Regulation 1924 / 2006, recognizes the role of zinc in normal cognitive functioning.

[0063] Examples of carotenoids include lutein, zeaxanthin and mixtures thereof.

[0064] Examples of omega 3 fatty acids include docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) and mixtures thereof.

[0065] Long-chain omega-3 fatty acids such as docosahexaenoic acid (DHA) have a beneficial effect on cognitive functions at the preclinical level in aged mice. Its beneficial effects on neuroinflammation are well accepted today and are widely described in the literature. The impact of omega-3 on cognitive functions has also been shown in humans. A meta-analysis of 10 randomized trials showed the benefit of omega-3 on certain areas of cognition. in patients with non-pathological cognitive decline. Observational studies have linked DHA consumption to improved cognitive performance and / or a reduction in age-related cognitive decline. More recently, an interventional study in elderly subjects complaining of cognitive impairment showed that fish oil supplementation was associated with improved cognitive abilities.

[0066] Examples of natural oils include fish oils, vegetable oils, oils from microalgae and their mixtures.

[0067] According to one embodiment, the invention also relates to a food or drink comprising the composition according to the invention and at least one constituent described in Table 1.

[0068] Process for obtaining

[0069] The composition according to the invention can be obtained by any process making it possible to obtain a mixture comprising at least 0.15% of catechin and / or epicatechin by weight relative to the total weight of the mixture, at least 15 ppm of safranal by weight relative to the total weight of the mixture, the safranal concentration being measured by HPLC method and at least 80 ppm of crocetin in aglycone and / or glycosylated form. It can be the mixture of the different molecules in the desired proportions or the mixture of plant extracts comprising the different molecules in the desired proportions.

[0070] Preferably, the compositions according to the invention are obtained by any process making it possible to obtain at least 0.9% of catechin and / or epicatechin, at least 50 ppm of safranal by weight relative to the total weight of the mixture, the safranal concentration being measured by HPLC method and at least 200 ppm of crocetin in aglycone and / or glycosylated form.

[0071] According to one embodiment, the compositions according to the invention are obtained by mixing at least: - an extract of Vitis vinifera and / or an extract of Vaccinium angustifolium and an extract of Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera, Vaccinium angustifolium and Crocus sativus, and / or - an extract obtained from a mixture of Vitis vinifera and Vaccinium angustifolium and an extract of Crocus sativus.

[0072] The extracts can be obtained by any process making it possible to obtain a mixture comprising at least 0.15% of catechin and / or epicatechin by weight relative to the total weight of the mixture, at least 15 ppm of safranal by weight relative to the total weight of the mixture, the safranal concentration being measured by HPLC method and at least 80 ppm of crocetin in aglycone and / or glycosylated form.

[0073] A particularly suitable process for obtaining an extract of Vitis vinifera comprises the following steps: - water and / or ethanol extraction of Vitis vinifera, preferably of all the leaves, fruits, skin, seeds and wood of Vitis vinifera and more preferably of fruits, skin, seeds. The quantity of solvent (30% v / v to 96% V / V) used is between 2 and 10 times the mass of material used. The extraction time can be between 30 minutes and 24 hours and the extraction temperature between 20°C and 80°C. The raw materials used can be in dry, fresh, or frozen whole or ground forms; - separation of the water and / or ethanol solution from the solid matter, for example by centrifugal decantation or by pressing and filtration; - evaporation of ethanol by vacuum evaporation at a temperature, preferably lower than 60°C and at a pressure lower than 100 mbars; - membrane separation of the previously desolventized extract so as to preferentially select the proanthocyanidin monomers and oligomers (having a degree of polymerization between 2 and 10 inclusive) and to eliminate the flavanol polymers (> decamers), to obtain an extract characterized by a content of flavanol polymers of less than 0.5% and more preferably less than 0.1% by weight relative to the total weight of the polyphenols in the extract. This step can be carried out using a filtration membrane having a cut-off threshold of less than 15,000 daltons and more preferably less than 3,000 daltons. -purification of the extract on a chromatographic column with adsorption resins followed by elution using a hydroethanolic solution preferably between 60 and 80% v / v - evaporation of the solvent and vacuum concentration of the eluate rich in polyphenols at a temperature preferably lower than 60°C and a pressure lower than 100 mbar; - drying of purified extracts by atomization, vacuum oven or freeze-drying with or without support such as maltodextrin.

[0074] A particularly suitable method for obtaining an extract of Vaccinium angus-tifolium comprises the following steps: - extraction with water and / or ethanol of Vaccinium angustifolium from all the leaves and fruits, preferably from the marc of the fruits of Vaccinium angustifolium obtained after pressing the fruits and separating the juice. The quantity of solvent (30% v / v to 96% V / V) used is between 2 and 10 times the mass of material used. The extraction time can be between 30 minutes and 24 hours and the extraction temperature between 20°C and 80°C. The raw materials used can be in dry, fresh, or frozen forms; - separation of the water and / or ethanol solution from the solid matter by centrifugal decantation or by pressing and filtration; - evaporation of ethanol by vacuum evaporation at a temperature preferably lower than 60°C and a pressure lower than 100 mbar; - drying of extracts by atomization, vacuum oven or freeze-drying with or without support such as maltodextrin.

[0075] A process particularly suitable for obtaining an extract of Crocus sativus: - use of stigmas of Crocus sativus, - grinding using a pin mill, to a size of 250pm, - hydroalcoholic extraction with 60% v / v ethanol, at a rate of 50g of saffron per liter of hydroalcoholic solution, - impregnation on maltodextrin, introduced into the hydroalcoholic solution - heat treatment in an oven for 48 hours at 40°C, - dosage of the extract molecules by UHPLC method.

[0076] The dosage of the extract by UHPLC method can be carried out as follows: a. Sample preparation: Extraction of the sample with a hydroalcoholic solution. Magnetic stirring for at least 1 hour then placed in an ultrasonic bath for at least 5 minutes. Membrane filtration then injection b. HPLC Elution: Binary Gradient: Solvent A (formic acid in MeCN) and Solvent B (formic acid in water) c. Detection: Crocins 440 nm, Picrocrocin derivatives 250 nm, Flavonoids 350 nm, Safranal 310 nm. d. Standards: Trans-crocin-4-gentiobiose-gentobiose, Trans-crocin 3-gentiobiose-glucose, beta-cyclocitral, Kaempferol glucoside, Safranal.

[0077] Once all the extracts have been obtained, the process for preparing the composition may comprise mixing the extract obtained from Vitis vinifera, Vaccinium angus-tifolium and Crocus sativus before or after the drying step, preferably after.

[0078] According to one variant, the method consists of implementing the following steps: a. mixture of Vitis vinifera and Vaccinium angustifolium extraction with water and / or ethanol of Vaccinium angustifolium, preferably of Vaccinium angustifolium fruits. The quantity of solvent (30% v / v to 96% V / V) used is between 2 and 10 times the mass of material used. The extraction time can be between 30 minutes and 24 hours and the extraction temperature between 20°C and 80°C. The raw materials used can be in dry, fresh, or frozen forms; b. separation of the water and / or ethanol solution from the solid marc by centrifugal decantation or by pressing and filtration; c. evaporation of ethanol by vacuum evaporation at a temperature preferably lower than 60°C and a pressure lower than 100 mbars; d. drying the extract by spraying or sublimation with or without a carrier such as maltodextrin.

[0079] Whatever the variant of the process, before the drying step, the process may comprise the following steps: a. loading onto a resin the mixed or unmixed extract solutions, b. rinsing the resin with water, c. application of a water / ethanol eluent solution to the resin, d. recovery of the purified eluate, e. evaporation of ethanol from said eluate, f. concentration of said eluate, g. drying said purified aqueous extract.

[0080] Use of the composition

[0081] The invention also relates to the composition according to any of the preceding embodiments for its use as a medicament or nutritional product, preferably for acting on cognitive and executive functions in a healthy human or animal but also in a sick human or animal, preferably suffering from a pathology associated with cognitive decline.

[0082] Cognitive decline is characterized by an age-related decline in cognitive and executive functions, particularly concentration, work, short- and long-term memory, reasoning abilities, judgment, problem-solving, and information processing speed. These impairments can lead to a decrease in self-esteem and quality of life. Age-related cognitive decline is the term used to describe the non-pathological form of deterioration in memory and cognitive functions that occurs as a result of the aging process within normal limits, taking into account a person's age. It is a complex process, with first signs emerging in humans between the ages of 35 and 65, without specific neurodegenerative lesions.Progressive cognitive decline is noticeable by the appearance of minor cognitive problems that affect 15 to 20% of the population aged 65 or older, and represents an unstable state. However, certain pathological forms can occur in addition to this "normal" cognitive decline. Among these pathologies, Alzheimer's disease is the most common cause of dementia, affecting more than 24 million people worldwide. It is irreversible in our current state of knowledge, the only available treatments being purely symptomatic. In animals, these pathologies can present in very similar ways. In dogs, for example, cognitive dysfunction syndrome (or CDS) is a widespread pathology characterized by disorientation. spatio-temporal, a loss of basic learning which often leads to uncleanliness, an alteration of sleep-wake cycles, an alteration of social interactions.

[0083] The composition according to the invention is capable of improving cognitive and executive functions in humans or animals. The combination of the three raw materials and the specificity of the extracts according to the invention comprising molecules combined in particular quantities, produces a synergistic effect in comparison with molecules taken in isolation or with existing extracts comprising these molecules in different proportions and quantities. The synergy relates to the neuroprotective effect and / or to the improvement of cognitive and / or executive functions in humans or animals.

[0084] Thus, according to a first embodiment, the composition is intended to be used in neuroprotection, improvement of cognitive functions, memory and / or executive functions, and attenuation of cognitive decline.

[0085] Preferably, the composition according to the invention can be used in healthy humans or animals, to improve cognitive functions and / or executive functions, and / or to limit non-pathological cognitive decline linked to age, preferably in a nutritional composition or a food supplement. It can in particular be used in healthy humans or animals to improve memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or intelligence and / or language and / or mood and / or stress and / or anxiety and / or vision and / or sleep and / or endothelial function and / or digestive tone and / or cholesterolemia and / or blood sugar.

[0086] According to a second embodiment, the composition is intended to be used in the prevention and / or treatment of pathologies associated with cognitive decline.

[0087] Preferably, the pathologies associated with cognitive decline are: - neurodegenerative diseases such as Alzheimer's disease, and / or Parkinson's disease, and / or Huntington's disease, and / or motor neuron diseases and / or age-related macular degeneration (AMD), -pathological cognitive decline and / or dementia and / or depression and / or anxiety and / or schizophrenia and / or mental retardation and / or cognitive dysfunction syndrome (CDS), -cardiovascular diseases and / or cerebrovascular diseases and / or diabetes and / or diseases of the circulatory system and / or inflammatory or non-inflammatory digestive diseases and / or disorders linked to the post-menopausal state in women.

[0088] According to a particular embodiment of the invention, the human or animal is elderly. Preferably, the elderly human or animal is a human or animal who has reached at least 50% of the average lifespan associated with his species.

[0089] The composition may be in any form suitable for administration oral in humans or animals, such as a form chosen from tablets, capsules, soft capsules, gel capsules, powders, solutions, microcapsules, sticks, suspensions, emulsions, food supplements, nutraceuticals, drinks, foods, bars, dairy products, powders for swallowing or rehydration, gels, oils, jams, sweets, chewing gums, soft drinks, dry drinks for rehydration, gelled drinks, compotes, soups, purees, desserts, entremets, ice creams / sorbets, pastries, preparations for gelling and / or thickening, ready meals, preparations for cereals, snacks, oral nutritional supplements, foodstuffs intended for special medical purposes, preferably in powder form.

[0090] For example, when it is a composition intended to be used as a nutritional product in humans, the composition may be in the form of food supplements, bars, dairy products, powders to be swallowed or rehydrated, gels, jams, candies, carbonated or non-carbonated drinks, dry drinks to be rehydrated or compotes.

[0091] It may also be a composition intended for the animal as a nutritional product such as, for example, dry foods, such as kibble (extruded, co-extruded or freeze-dried), treats, snacks, wet or semi-wet foods such as pieces in sauce, pieces in jelly, drinks, or even food supplements. Preferably, the composition is integrated into dry foods such as kibble. The term "animal" means any animal that can receive a composition according to the invention, for example, but not limited to, a pet, poultry, a pig, a ruminant, a goat, or even a mouse. Preferably, the animal is a pet, such as a cat or a dog. More preferably, the animal is a dog.

[0092] For example, when it is a composition intended to be used as a medicament in humans, the composition may be in the form of tablets, capsules, soft capsules, gelcaps, powders, solutions, microcapsules, sticks, syrups, suspensions or emulsion.

[0093] Finally, it may be a medicine intended for animals, or a veterinary product, such as tablets, capsules, sprays, or liquids administered by drops.

[0094] Advantageously, the mixture of molecules intended for the animal can be integrated into a composition, in particular into a nutritional or therapeutic composition, by inclusion, that is to say by adding it into the mass of the composition for example by impregnation or mixing, or by coating, that is to say by applying it to the surface of the composition, by spraying or by dusting, for example by mixing it pre-mixing with one or more ingredients such as at least one palatability factor.

[0095] Preferably, the composition according to the invention is delivered at a daily dose allowing administration in humans: - at least 0.05 mg per kg of body weight of catechin and / or epicatechin, - at least 0.5 pg per kg of body weight of safranal and / or - at least 2.5 pg per kg of body weight of crocetin in aglycone and / or glycosylated form.

[0096] Even more preferably, the composition according to the invention is delivered at a dose allowing administration in humans: - at least 0.1 mg per kg of body weight of catechin and / or epicatechin, - at least 1 pg per kg of body weight of safranal and / or - at least 5 pg per kg of body weight of crocetin in aglycone and / or glycosylated form.

[0097] According to another embodiment, the composition according to the invention is delivered at a dose allowing administration in dogs: - at least 0.02 mg per kg of body weight of catechin and / or epicatechin, and - at least 0.1 pg per kg of body weight of safranal, and / or - at least 1 pg per kg of body weight of crocetin in aglycone and / or glycosylated form.

[0098] Preferably, the composition according to the invention is delivered at a dose allowing administration in dogs: - at least 0.04 mg per kg of body weight of catechin and / or epicatechin, and - at least 0.2 pg per kg of body weight of safranal, and / or - at least 2 pg per kg of body weight of crocetin in aglycone and / or glycosylated form.

[0099] According to another embodiment, the composition is intended to be contained in a food and further comprises: - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least 1.8 pg of vitamin E, at least 8.2 pg of selenium, at least 1.5 mg of zinc provided per 100 g or 100 ml of non-drink products, or - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least 1.8 pg of vitamin E, at least 8.2 pg of selenium, at least 1.5 mg of zinc provided per recommended dose of food supplements, or - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least at least 1.8 pg of vitamin E, at least 8.2 pg of selenium, at least 1.5 mg of zinc provided per portion if the package contains only one portion, or - at least 60 pg of vitamin A, at least 15 pg of vitamin B9, at least 0.1 pg of vitamin B12, at least 6 mg of vitamin C, at least 0.3 pg of vitamin D, at least 0.9 pg of vitamin E, at least 4.1 pg of selenium, at least 0.7 mg of zinc per 100 ml of drinks, or - at least 40 mg of eicosapentaenoic acid and / or docosahexaenoic acid per 100 g or 100 kcal.

[0100] For the purposes of the invention, the term "recommended dose of food supplements" means 3630 mg / day. The term "portion" means, for example, 90g or 100g of compote in a pot or pouch, 125g of yogurt or dessert cream, 40g of biscuits or madeleines, 200ml of soup or milk drink, 70g of dehydrated soup in a sachet, 12g of powder containing the composition according to the invention to be added to a food preparation, 5 enriched sweets or 180g of a food dish prepared in a tray.

[0101] In particular, the composition is contained in a food and is delivered at a daily dose allowing administration in humans: - at least 15% of the reference nutritional values ​​of vitamins and trace elements provided per 100 g or 100 ml of non-drink products, preferably 30%, even more preferably 100%, and / or - at least 15% of the nutritional reference values ​​of vitamins and trace elements per recommended dose of food supplements, preferably 30%, even more preferably 100%, and / or - at least 7.5% of the nutritional reference values ​​of vitamins and trace elements per 100 ml of drinks, preferably 15%, even more preferably 100% and / or - at least 15% of the nutritional reference values ​​of the vitamins and trace elements provided per portion if the packaging contains only one portion, preferably 30%, even more preferably 100% and / or - at least 40 mg of eicosapentaenoic acid and / or docosahexaenoic acid per 100 g or 100 kcal, preferably 80 mg.

[0102] By reference nutritional values ​​(NRV) we mean the values ​​indicated in Table 1 below.

[0103] [Tables 1] Constituting 7.5% of the NRV 15% of the NRV 30% of the NRV 100% of the NRV Vitamin A (pg) 60 120 240 800 Vitamin B9 (pg) 15 30 60 200 Vitamin B12 (pg) 0.1 0.3 0.7 2.5 Vitamin C (mg) 6 12 24 80 Vitamin D (pg) 0.3 0.7 1.5 5 Vitamin E (mg) 0.9 1.8 3.6 12 Selenium (pg) 4.1 8.2 16.5 55 Zinc (mg) 0.7 1.5 3 10

[0104] Such a composition is also intended to be used according to one of the uses described above.

[0105] According to another embodiment, the invention also relates to the non-therapeutic use of a composition according to the invention in healthy humans or animals, to improve cognitive functions and / or executive functions, and / or to limit normal age-related cognitive decline.

[0106] Preferably, the non-therapeutic use is intended to improve memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or language and / or mood and / or stress and / or anxiety and / or vision and / or sleep and / or endothelial function and / or digestive tone and / or cholesterolemia and / or blood sugar.

[0107] Said composition intended to be used in a non-therapeutic manner is administered at a dose described above.

[0108] The invention is now illustrated through examples and test results demonstrating the synergistic effect on cognitive functions of the composition which is the subject of the present application. Examples

[0109] Example of a composition intended for humans.

[0110] Example 1: Composition in powder form according to the invention

[0111] Example 1 is a 630 mg composition comprising: - 600mg of a mixture of molecules including 0.9% Catechin / Epicatechin - 30mg of a mixture of molecules including 90 ppm of Safranal and 400 ppm of aglycone and glycosylated crocetins.

[0112] The composition is obtained by mixing the constituents under the conditions conventional compositions known to those skilled in the art. Alternatively, the composition may be obtained from an extract or mixture of extracts. Said composition according to Example 1 is in powder form and may serve as a base for the formulations described below.

[0113] Example 2: Composition in powder form according to the invention

[0114] Example 2 is a 630 mg composition comprising: - 600mg of a mixture of molecules comprising 21% Catechin / Epicatechin, and - 30mg of a mixture of molecules comprising 2100 ppm Safranal and 1.1% aglycone and glycosylated Crocetins.

[0115] The composition is obtained by mixing the constituents under conventional conditions known to those skilled in the art and can also serve as a base for other formulations. Alternatively, the composition can be obtained from an extract or a mixture of extracts.

[0116] Example 3: Composition in powder form according to the invention

[0117] Example 3 is a 630 mg composition comprising: - 600mg of a mixture of molecules including 0.9% Catechin / Epicatechin, 180 ppm Resveratrol, 4 ppm Ferulic Acid, 45 ppm Quercetin and derivatives, 470 ppm Anthocyanins and - 30mg of a mixture of molecules including 90 ppm of Safranal and 400 ppm of Crocin.

[0118] The composition is obtained by mixing the constituents under conventional conditions known to those skilled in the art and can also serve as a base for other formulations. Alternatively, the composition can be obtained from an extract or mixture of extracts.

[0119] Example 4: Composition in powder form according to the invention

[0120] Example 4 is a 3630 mg composition comprising: - 630mg of a mixture according to example 3 and - 3g of a mixture comprising 1g of omega 3 fatty acids, 20mg of vitamin E, 60mg of vitamin C, 25mg of Polygonum extract and 20mg of olive leaf extract.

[0121] The composition is obtained by mixing the constituents under conventional conditions known to those skilled in the art.

[0122] Example 5: Composition in capsule form according to the invention

[0123] Example 5 is a 400 mg capsule, consisting of: -300mg of a mixture of Vitis vinifera and Vaccinium angustifolium extract including 5% catechin and / or epicatechin, - 15mg of Crocus sativus extract comprising 0.2% safranal and 1% aglycone and glycosylated crocetins, - 85 mg of maltodextrin.

[0124] The composition is obtained by mixing the constituents under conventional conditions known to those skilled in the art, and placed in a capsule under conventional conditions as well.

[0125] The recommended dosage is 1 to 2 capsules per day.

[0126] Example 6: Nutritional composition intended for humans,

[0127] Example 6 is a 400 mg capsule, consisting of: -300mg of a mixture of Vitis vinifera and Vaccinium angustifolium extract including 1% catechin and / or epicatechin, -15mg of a Crocus sativus extract comprising 0.2% safranal and 1% aglycone and glycosylated crocetins, -200 pg of vitamin B9, 50 pg of vitamin B12, 7.5 pg of vitamin D, 2.5 mg of zinc and 50 mg of maltodextrin.

[0128] The composition is obtained by mixing the constituents under conventional conditions known to those skilled in the art and placed in a capsule under conventional conditions as well.

[0129] The recommended dosage is 1 to 2 capsules per day.

[0130] Example 7: Nutritional composition intended for humans,

[0131] Example 7 is a capsule consisting of the formulation according to Example 6 and a soft capsule containing 1g of fish oil rich in EPA / DHA, 12mg of lutein and 20mg of vitamin E.

[0132] The recommended dosage is 1 to 2 capsules per day.

[0133] Example 8: Composition in capsule form.

[0134] Example 8 is a capsule consisting of the formulation according to Example 6 and 25 mg of flavanones from a citrus extract, 0.5 mg of oleuropein from an olive leaf extract and 10 mg of resveratrol.

[0135] The recommended dosage is 1 to 2 capsules per day.

[0136] Example 9: Composition in capsule form.

[0137] Example 9 is a capsule consisting of the formulation according to Example 8 and a caper extract.

[0138] Example 10: Nutritional drink intended for humans

[0139] Example 10 is a 50 ml drink containing the equivalent of 300 mg of a mixture of Vitis Vinifera and Vaccinium angustifolium extract and 15 mg of Crocus sativus extract. The extracts may be microencapsulated.

[0140] The recommended dosage is 1 to 2 shots per day.

[0141] Example 11: Nutrient-enriched oil intended for humans

[0142] Example 11 is a vegetable oil consisting of olive oil and rapeseed oil containing polyphenols from an extract of Vitis Vinifera and Vaccinium angustifolium and carotenoids from an extract of Crocus sativus. A daily dose 20ml contains 600mg of a blend of Vitis Vinifera and Vaccinium an-gustifolium extract and 30mg of Crocus sativus extract.

[0143] Example 12: High-protein powder preparation

[0144] Example 12 is a food formulation for special medical purposes or a protein-enriched food formulation. The composition is detailed in Table 2. The dosage may be, for example, a daily dose of said powdered preparation. Said dose may be diluted in a drink or dish and represents approximately 12g.

[0145] Example 13: Yogurt

[0146] Example 13 is a formulation in the form of yogurt, to be consumed 2 to 3 times per day. The composition is detailed in Table 2.

[0147] Example 14: Preparation of fruit on sugar

[0148] Example 14 is a fruit preparation on sugar to be consumed in 1 to 2 portions on bread / rusks for breakfast or as a snack. The composition is detailed in Table 2.

[0149] Example 15: Dehydrated preparation for soup

[0150] Example 15 is an individual dehydrated preparation for high-protein soup of 70g to be diluted using a spoon in 130 to 200 ml of hot water depending on the desired texture and consume 1 to 3 preparations per day. The composition is detailed in Table 2.

[0151] [Tables 2] Composition according to the invention Example 12 Example 13 Example 14 Example 15 Portion (g) 12 100 125 10 100 70 100 Constituents Catechin+epicatechin (mg) 6 50 6 6 50 6 9 Safranal (mg) 0.1 1 0.1 0.1 1 0.1 0 Crocin (mg) 0.3 3 0.3 0.3 3 0.3 0 Other Proteins (g) 10 83 10 10 1000 10 14 Vitamin D (pg) 15 125 15 15 1500 10.5 15 Vitamin E (mg) 20 167 20 20 2000 20 29 Vitamin B9 (pg) 400 3333 400 400 40000 400 571 Vitamin B12 (pg) 100 833 100 100 10000 100 143 Vitamin C (mg) 60 500 60 60 6000 60 86 Zinc (mg) 5 42 5 5 500 5 7

[0152] Example 16: Dog kibble

[0153] Example 16 is a formulation in the form of kibble intended for a 10 kg dog fed with 200 g of kibble / day comprising: - 0.04 mg per kg of body weight of catechin and / or epicatechin, and - 0.2 pg per kg of body weight of safranal, and / or - 2 pg per kg of body weight of crocin.

[0154] EVALUATION OF THE EFFECTIVENESS OF THE COMPOSITION ACCORDING TO THE INVENTION

[0155] Test 1: Evaluation of the synergistic effect on acetylcholinesterase activity.

[0156] The objective of this test is to compare the activity of an enzyme, acetylcholinesterase (AchE) (enzyme responsible for the degradation of acetylcholine) in the presence of the composition according to the invention with the activity of acetylcholinesterase in the presence of polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium or carotenoids contained in an extract of Crocus sativus.

[0157] AchE activity was monitored and measured in vitro by a colorimetric reaction following the Ellman method. Acetylthiocholine (ATCI) is degraded to thiocholine and acetic acid by AChE. Thiocholine then reacts with the dithiobisnitrobenzoic ion (DTNB) and the product ion (TNB) gives a yellow color. In a 96-well plate, the following quantities were deposited in each well: 55pl buffer (Tris-HCl 50mM / 0.1% BSA), 75pl DTNB (5mM), 25pl AChE (0.22U / ml), 20pl of test sample, Galantamine GNT) (5pM), (known AChE inhibitor) or water.

[0158] In order to study the synergistic effect, the test was carried out using a mixture of polyphenols comprising both Catechin / Epicatechin at 10pM, Resveratrol at 25nM and Ferulic Acid at 2.33pM, a mixture of Carotenoids comprising Safranal at 35.3pM and Crocetin at 162.17pM and a composition according to the invention comprising Catechin / Epicatechin at 10pM, Resveratrol at 25nM, Ferulic Acid at 2.33pM, Safranal at 35.3pM and Crocetin at 162.17pM.

[0159] At the last moment, 25 μL of ATCI (ImM) was added to initiate the reaction. Subsequently, absorbance monitoring at 405 nM was carried out for 45 min. The results were analyzed as follows. For each well, the absorbance values ​​over time (Ax) are normalized with respect to the first absorbance measurement (ATO) according to the following formula: Normalized absorbance (An) = Ax - ATO. The percentage of inhibition of AchE was determined on the normalized absorbances, at T = 40 min, according to the following equation [Math 1].

[0160] [Math.l] (4n Control — An Blank) — (An Sample — An Blank sample) % inhibition = ------------------------------------------------------------------- X100 (4n Sample — An Blank Sample)

[0161] Where An Control is the average of the normalized absorbance measurements of the normal reactions (buffer+DTNB+AchE+H2O), An Blank is the average of the normalized absorbance measurements of the reagents without the presence of enzyme (buffer+DTNB+H2O), An Sample is the normalized absorbance of the reaction in the presence of the sample (buffer+DTNB+AchE+sample), An Sample Blank is the average of the normalized absorbance measurements of the reagents in the presence of the sample (buffer+DTNB+sample).

[0162] The absorbance values ​​(at T=40min) were statistically compared using a two-way analysis of variance (ANOVA) where factor 1 was the treatment with polyphenols from Vitis vinifera, and Vaccinium angustifolium and factor 2 was the treatment with carotenoids from Crocus sativus. A value of p<0.05 is considered significant.

[0163] In the presence of polyphenols from Vitis vinifera extract, and Vaccinium angustifolium, a small but significant decrease (5.3%) in AchE activity was observed. In the presence of carotenoids from Crocus sativus extract, a di A significant decrease (21.6%) in AchE activity was also observed. Finally, a significant interaction was found between polyphenols from Vitis vinifera and Vaccinium angustifolium extracts and carotenoids from Crocus sativus extracts, in particular between the molecules of the composition according to the invention, namely catechins and epicatechins, safranal and crocetin. Thus, such a composition according to the invention showed a decrease (44.8%) in AchE activity greater than that observed with a mixture of polyphenols or carotenoids. This decrease (44.8%) is greater than that of the cumulative polyphenols and carotenoids (26.9%).

[0164] Such results demonstrate that there are synergistic interactions between the polyphenols derived from Vitis vinifera extract, and from Vaccinium angustifolium and the carotenoids derived from Crocus sativus extract, in particular between the molecules of the mixture according to the invention, namely catechins and epicatechins, safranal and crocetin, on the activity of AchE by making it possible to reduce its activity. The reduction in its activity could thus make it possible to improve cholinergic transmission and consequently cognitive functions. Such a synergistic effect of the composition of the invention thus makes it possible to propose a formulation making it possible to improve cognitive functions.

[0165] Trial 2: Evaluation of the synergistic effect on neuroprotection

[0166] This test aims to compare, following exposure to glutamate (at a concentration neurotoxic), neuronal survival in the presence of the composition according to the invention with neuronal survival in the presence of polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium or carotenoids contained in an extract of Crocus sativus.

[0167] In order to study the synergistic effect, the test was carried out using a mixture of polyphenols comprising both Catechin / Epicatechin at 3pM, Resveratrol at 7.5nM and Ferulic Acid at 0.7pM, a mixture of Carotenoids comprising Safranal at 10pM and Crocetin at 48.7pM and a composition according to the invention comprising Catechin / Epicatechin at 3pM, Resveratrol at 7.5nM, Ferulic Acid at 0.7pM, Safranal at 10pM and Crocetin at 48.7pM

[0168] To do this, primary cultures of cortical neurons from rats were prepared in 96-well plates (seeding at 25,000 cells / well). After 10 days of culture, the composition according to the invention, the polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium or the carotenoids contained in an extract of Crocus sativus were introduced into the culture medium for a period of 3 days. On the 13th day of culture, the cortical neurons were exposed to Glutamate for 20 min. Glutamate was added at the final concentration of 20 pM in the presence of the composition according to the invention, the polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium or carotenoids contained in an extract of Crocus sativus. After the 20 min incubation, the glutamate was removed and replaced with culture medium with the composition according to the invention, polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium or carotenoids contained in an extract of Crocus sativus for an additional 48 hours. 48 hours after exposure to Glutamate, the cells were washed with phosphate buffer and fixed with ethanol (95%) and acetic acid (5%) for 5 minutes at -20°C. Then immunofluorescence staining of microtubule-associated proteins (MAP-2) was performed. This antibody specifically recognizes neurons and the neuritic network, thus allowing the study of neuronal survival. For each condition, 30 photos per well were taken and analyzed with the same parameters. The number of surviving neurons was then quantified.For each condition, neuronal survival was expressed as a percentage relative to the control condition. Differences in neuronal survival were statistically compared using analysis of variance (ANOVA) followed by Fisher's test. A p-value <0.05 is considered statistically significant.

[0169] In the presence of glutamate, neuronal survival is significantly lower than the control (66% vs 100%). Glutamate therefore induced a neurotoxic effect. In the presence of polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium, glutamate also induced neuronal death. The polyphenols contained in an extract of Vitis vinifera and Vaccinium angustifolium therefore did not show a neuroprotective effect. In the presence of carotenoids contained in an extract of Crocus sativus, a significant increase in neuronal survival (83%) following the addition of glutamate was observed. Finally, in the presence of the composition according to the invention, a strong and significant increase in neuronal survival (92%) was observed.Thus, in the presence of the composition according to the invention, neuronal survival is greater than that observed in the presence of a mixture of polyphenols or carotenoids, thus demonstrating that there are synergistic interactions between the polyphenols derived from extract of Vitis vinifera and Vaccinium angustifolium and the carotenoids derived from extract of Crocus sativus. Such a synergistic effect of the composition of the invention thus makes it possible to propose a neuroprotective formulation and therefore to improve cognitive functions.

Claims

Claims

1. Composition comprising a mixture of at least one extract of Vitis vinifera, at least one extract of Vaccinium angustifolium and at least one extract of Crocus sativus.

2. Composition according to the preceding claim, characterized in that said mixture comprises a mixture of molecules comprising: - at least 0.15% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, - at least 15 ppm (parts per million in the mixture) of safranal by weight relative to the total weight of the mixture measured by HPLC method, and - at least 80 ppm of crocetin in aglycone and / or glycosylated form

3. Composition according to the preceding claim, characterized in that the mixture comprises at least 0.9% of catechin and / or epicatechin, the percentage being given by weight relative to the total weight of the mixture, preferably at least 15%.

4. Composition according to one of the preceding claims, characterized in that the mixture comprises at least 50 ppm of safranal by weight relative to the total weight of the mixture measured by HPLC method, preferably at least 90 ppm.

5. Composition according to one of the preceding claims, characterized in that the mixture comprises at least 200 ppm of crocetin in aglycone and / or glycosylated form, preferably at least 1000 ppm of crocetin in aglycone and / or glycosylated form.

6. Composition according to one of the preceding claims, characterized in that the mixture further comprises at least 0.5 ppm of ferulic acid, preferably at least 4 ppm.

7. Composition according to one of the preceding claims, characterized in that the mixture also comprises: - at least 30 ppm of resveratrol, and / or - at least 5 ppm of quercetin and / or quercetin glycosides, and / or - at least 50 ppm of aglycone and / or glycosylated anthocyanins.

8. Composition according to one of the preceding claims in that the mixture of Vitis vinifera and Vaccinium angustifolium comprises at least 1% of catechin and / or epicatechin of the total weight of said extract, and 5 ppm of ferulic acid.

9. Composition according to one of the preceding claims in that the mixture of Vitis vinifera and Vaccinium angustifolium comprises at least 200 ppm of resveratrol, 50 ppm of quercetins and / or quercetin glycosides and / or at least 500 ppm of anthocyanins.

10. Composition according to one of the preceding claims, characterized in that the Crocus Sativus extract has a safranal content of at least 0.2% of said extract.

11. Composition according to one of the preceding claims, characterized in that the Crocus Sativus extract contains at least 1% of aglycone and / or glycosylated crocetin, 0.5% of picrocrocin, and 500 ppm of flavonoids derived from kaempferol of the total weight of said extract.

12. Composition according to one of the preceding claims, characterized in that the mixture also comprises at least one other constituent chosen from antioxidants, natural extracts, vitamins, trace elements, carotenoids, omega 3 fatty acids, natural oils, and mixtures thereof.

13. Composition according to the preceding claim, characterized in that the antioxidants are chosen from polyphenols, vitamin C, E, A, zinc, selenium and mixtures thereof; the plant extracts are chosen from extracts of coffee, olive or olive leaves, blackcurrant, polygonum, citrus fruits, marigold and mixtures thereof; the vitamins are chosen from vitamin B9, B12, C, D, E, A (including pro-vitamins A), their derivatives and mixtures thereof; the trace elements are chosen from zinc, selenium, chromium and mixtures thereof; the carotenoids are chosen from lutein, zeaxanthin and mixtures thereof; the omega 3 fatty acids are chosen from docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and mixtures thereof; Natural oils are chosen from fish oils, vegetable oils, oils from microalgae and their mixtures.

14. Composition according to one of the preceding claims, in which the composition is in a form chosen from tablets, capsules, soft capsules, gel capsules, powders, solutions, microcapsules, sticks, suspensions, emulsions, food supplements, nutraceuticals, drinks, foods, bars, dairy products, powders for swallowing or rehydrating, gels, oils, jams, candies, chewing gums, carbonated drinks, dry drinks for rehydration, gelled drinks, compotes, soups, purees, desserts, desserts, ice creams / sorbets, pastries, gelling and / or thickening preparations, ready meals, cereal preparations, finger foods, oral nutritional supplements, foodstuffs for special medical purposes.

15. Composition according to one of the preceding claims, for its use as a medicament or nutritional product for humans or animals.

16. Composition for its use according to the preceding claim, in neuroprotection, improvement of cognitive functions, memory and / or executive functions, attenuation of cognitive decline.

17. Composition for use according to claim 15, in the prevention and / or treatment of pathologies associated with cognitive decline.

18. Composition for its use according to the preceding claim, in the prevention and / or treatment of: - neurodegenerative diseases such as Alzheimer's disease, and / or Parkinson's disease, and / or Huntington's disease, and / or motor neuron diseases and / or age-related macular degeneration (AMD), - pathological cognitive decline and / or dementia and / or depression and / or anxiety and / or schizophrenia and / or mental retardation and / or cognitive dysfunction syndrome (CDS), - cardiovascular diseases and / or cerebrovascular diseases and / or diabetes and / or diseases of the circulatory system and / or inflammatory or non-inflammatory digestive diseases and / or disorders related to the post-menopausal state in women.

19. Non-therapeutic use of a composition according to one of claims 1 to 14 in healthy humans or animals, to improve cognitive functions and / or executive functions, and / or to limit normal age-related cognitive decline.

20. Non-therapeutic use according to the preceding claim, for improving memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or language and / or mood and / or stress and / or anxiety and / or vision and / or sleep and / or endothelial or erectile function, and / or digestive tone and / or cholesterolemia and / or blood sugar.

21. Food comprising the composition according to one of claims 1 to 14 and: - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least 1.8 mg of vitamin E, at least 8.2 pg of Selenium, at least 1.5 mg of zinc provided per 100 g or 100 ml of non-drink products or - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least 1.8 mg of vitamin E, at least 8.2 pg of Selenium, at least 1.5 mg of zinc provided per recommended dose of food supplements, or - at least 120 pg of vitamin A, at least 30 pg of vitamin B9, at least 0.3 pg of vitamin B12, at least 12 mg of vitamin C, at least 0.7 pg of vitamin D, at least 1.8 mg of vitamin E, at least 8.2 pg of Selenium, at least 1.5 mg of zinc provided per portion if the package contains only one portion, or - at least 60 pg of vitamin A, at least 15 pg of vitamin B9, at least 0.1 pg of vitamin B12, at least 6 mg of vitamin C, at least 0.3 pg of vitamin D, at least 0.9 mg of vitamin E, at least 4.1 pg of Selenium, at least 0.7 mg of zinc per 100 ml of drinks or - at least 40 mg of eicosapentaenoic acid and / or docosahexaenoic acid per 100 g or 100 kcal.

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