WO2013054267A1

WO2013054267A1 - Synergistic composition for preventing and treating disorders connected with ageing

Info

Publication number: WO2013054267A1
Application number: PCT/IB2012/055476
Authority: WO
Inventors: Giulia Federica Merizzi
Original assignee: Medestea Biotech S.P.A.
Priority date: 2011-10-10
Filing date: 2012-10-10
Publication date: 2013-04-18
Also published as: ITTO20110901A1

Abstract

Pharmaceutical or dietary supplement composition, comprising, as active ingredients with combined or synergistic effect, curcumin and astaxanthin, for use in the prevention and treatment of disorders connected with ageing of the organism.

Description

Synergistic composition for preventing and treating disorders connected with ageing Field of the invention The invention relates to a composition to be used in the area of the prevention and treatment of problems/disorders connected with general ageing of the body and with chronic disorders connected therewith, such as neurodegenerative disorders of the central and of the peripheral nervous system, cardiovascular diseases, ageing of the skin and metabolic disorders. In particular the invention relates to a mixture of compounds capable of specifically activating transcription factors connected with expression of a family of genes and proteins with anti-inflammatory and antioxidant activity, able to increase the defence and regeneration capacities of cells, of specific tissues and of the whole body.

Summary of the invention

The object of the invention is a formulation that can be administered by the oral route, without particular contraindications, whether in relation to the subjects to whom it can be administered, or in relation to the amounts administered, having an effect of preventing, with regular long-term use, delaying and partially preventing cellular degeneration due to ageing.

The formulation according to the invention and its use are defined in the appended claims.

This formulation is characterized in that it comprises, as active agents, curcumin or a mixture of curcuminoids, in combination with astaxanthin and optionally one or more vehicles or excipients for pharmaceutical, food or cosmetic use.

Curcumin (l,7-bis[-hydroxy-3-methoxyphenyl]-l,6-heptadiene-3,5-dione) is a compound belonging to the class of polyphenolic compounds. It is a yellow spice, extracted from the rhizome of Curcuma longa (Zingiberaceae), a perennial herb, widely cultivated in Asia. It is commonly used as a flavouring and colouring agent in foodstuffs. Its anti-inflammatory and tumour-preventing properties are well known and more recently its use as neuroprotective agent has been proposed (cf. WO2004/075883).

The term curcuminoids comprises curcumin, whether natural or synthetic, and its derivatives, such as in particular demethoxycurcumin and bisdemethoxycurcumin, naturally present in extracts of Curcuma longa.

In the formulations according to the invention, the extract of Curcuma longa constitutes the preferred source of curcumin or curcuminoids.

Astaxanthin is a reddish-orange pigment naturally present in aquatic animals. Chemically it is a xanthophylline, belonging to the carotenoid family, of formula 3,3'-dihydroxy-[P],[$]'- carotene-4,4'-dione. Owing to its structural characteristics, astaxanthin displays high antioxidant activity.

In the formulations according to the invention, astaxanthin can be used both in the form of the pure compound, or preferably as an extract obtained for example from the alga Haematococcus pluvialis.

The extract of Curcuma longa and astaxanthin have demonstrated, unexpectedly, the capacity for activating, synergistically and specifically, the system of the transcription factor Nrf2 and the related genes, increasing the cellular defences. Moreover, their capacity for activating mitochondrial biogenesis has been demonstrated in various cells and tissues. Owing to this action, the formulation according to the invention is effective in promoting the vitality and the functions of the body's cells and tissues and is particularly effective in protecting endothelial cells, skin cells and cells of the nervous system from deterioration and from death related to degenerative processes and to ageing in general. Prior art

According to the most recent modern theory that aims to explain the biological phenomenon of ageing, a number of mechanisms that are partly endogenous (developing inside the body) and partly exogenous (environmental) cause cellular changes in the course of life (for example in the cell membrane, in enzymes, in proteins, in DNA and RNA).

These changes would quickly lead to ageing and death if our body did not possess appropriate defence mechanisms, the efficiency of which is genetically controlled. Although there are various mechanisms that lead to oxidative stress and to inflammation, numerous studies suggest an important role for the signalling mechanisms mediated by the nuclear transcription factor E2-related factor 2 (Nrf2) in the activation of cellular protection. Nrf2 is a member of the Cap'n'Collar family of transcription factors, and is sequestered in the cytoplasm by binding to the protein Kelch ECH-associated protein 1 (Keapl) in non-stimulated conditions. Keapl plays a central role in regulation of the response of Nrf2.

In normal conditions, Nrf2 is the target of Keapl, which promotes proteasome degradation of Nrf2 by means of interactions with ubiquitin ligase. Keapl further functions as a sensor of stress signals, by stress-induced oxidation of key residues of cysteine that lead to variations in conformation and to inability to bind Nrf2. Various stimuli, including oxidative stress and electrophilic compounds, such as polyphenols, lead to destruction of the Nrf2/Keapl complex, releasing Nrf2 for translocation to the nucleus, where it interacts with leucine zipper transcription factors, such as members of the Maf and Jun family and binds to a cis-acting element, the antioxidant response element (ARE, also designated EpRE or OSRE).

Nrf2-ARE is a primary pathway that regulates phase 2 antioxidant responses, triggering the simultaneous expression of numerous protective and scavenger enzymes. ARE is present in various inducible cytoprotective genes, correlated with the so-called cellular stress response, such as glutathione-S-transferase, haem oxygenase 1 (HO-1), glutamate cysteine ligase, ferritin, γ-glutamylcysteine synthetase, NAD(P)H quinone oxidoreductase thioredoxin and periredoxins, and this emphasizes Nrf2 with the central node in the cell survival response.

By constant activation of this complex genetic machinery, it is thus possible gradually to repair any damage as it arises. Based on this evidence, the modulation of Nrf2 and HO-1 in a different context, including the brain, the skin and vessels, has recently been suggested as a potential pharmaceutical strategy for the treatment and prevention of degenerative disorders and pathologic ageing. Many epidemiological studies in the past have indicated a direct correlation between the type of diet and the incidence and severity of chronic diseases linked with ageing, thus giving rise to the idea that by stimulating the activity of genes and proteins, chemical compounds contained in foods can influence the equilibrium between the healthy condition and the diseased condition in an individual and thus influence the quality of ageing, well- being and the beginning of degenerative diseases.

Detailed description of the invention

The invention therefore relates to a formulation that comprises a combination of curcumin or curcuminoids, optionally in the form of extract of Curcuma longa, and astaxanthin or an extract containing it.

It was discovered that the formulation according to the invention, when used regularly, gives rise to a prophylactic, protective effect on the degenerative processes induced by ageing.

Moreover, this protective effect is more effective if administration is begun before the start of the ageing process. Therefore the use of the formulation of the invention is recommended for individuals of any age, not only for elderly individuals.

The diseases or disorders associated with skin ageing that can be treated using the compositions according to the invention include, but are not limited to, skin damage through photo-ageing, dry or scaly skin, thinning of the skin or of the epidermis, actinic elastosis, wrinkles, crow's feet, liver spots and/or ageing spots, hyperpigmentation of the skin, loss of coloration, fragility, reduced skin tone, lowered irritation threshold, loss of tautness and sagging, loss of elasticity, laxity, reduced flexibility of the skin, loss of epidermal water resulting in loss of moisture.

The compositions according to the invention can be aimed at therapeutic, cosmetic and/or cosmetic/therapeutic use. Their administration results in prevention, reduction or delay for example of the formation of wrinkles, in prevention, reduction or delay in loss of muscle tone and of the formation of skin blemishes. The compositions are moreover able to moderate and assist in the maintenance of smooth, firm skin with improved elasticity.

The extract of Curcuma longa and astaxanthin are undoubtedly among the most interesting bioactive natural compounds for which anti-ageing properties have recently been suggested, with in vivo and in vitro data reported in various scientific publications.

The invention is based on the identification, by the applicant, of a synergistic effect between the two active compounds in relation to specific biological parameters that are measurable both at cellular level, and at the level of the organism.

Before presenting the results obtained, it is necessary to define what is meant in the present description by "pharmacologic synergism" and by "cellular parameters directly relevant in relation to cellular ageing". In this connection, the scientifically most rigorous definition of biological synergism is adopted, which derives from investigation of the action of drugs. Pharmacologic synergism occurs when two drugs interact to increase or amplify one or more effects, or their side- effects. In other words, two drugs that clearly produce two similar effects in some cases produce exaggerated or reduced effects when used concomitantly and a quantitative evaluation is necessary to distinguish these cases from simple additive action (Tallarida R.J. Drug synergism: its detection and applications, J. Pharmacol. Exp. Ther. September 2001 ; 298(3): 865-72). The biological effects that we are focusing on are positive modulation of cellular defence and repair systems, with a marked effect on cell survival and on longevity, both in terms of the single cell and cellular populations.

Firstly it must be borne in mind that the pharmacologic definition of synergism, when translated in terms of molecular events, often refers to the capacity of two bioactive substances, which generate synergism, to increase the cellular functions, eliciting pathways that are originally separate, but that converge at higher levels in the elementary cellular activities that contribute to the same primary cellular programme.

In other words, if there are two ways for a cell to avoid death, controlled by two pathways that are uncorrected or minimally correlated A and B, increase of only A or only B by two compounds will probably produce saturation of the pathways at the higher doses and therefore a combined effect that is less than the additive effect.

Complete stimulation of A and B, however, might produce additivity and even synergism (a) since all the activities that lead to that response are engaged, (b) since the interactions that occur when two pathways converge can become mutually potentiating or, finally, (c) since synchronous activation of both allows the cell to move towards discrete, but sequential phases of activity.

The model used by the applicant made it possible to determine that curcumin, or extract of Curcuma longa, and astaxanthin perform a synergistic action in preventing the damage induced by ROS (Reactive Oxygen Species) and more generally the damage linked to cell death, since each of these activates a pathway of cellular response to the damage.

In more detail, these active agents display a specific capacity for synergistic activation of the nuclear transcription factor Nrf2. Nrf2 interacts with the sequence of ARE at the level of promoter of various inducible genes, endowed with critical properties of defence and repair, resulting in cellular overexpression of these genes, an increase of cellular defences and inhibition of cell death. It has been demonstrated that many electrophilic polyphenols activate the Nrf2-ARE pathway, including the extract of Curcuma longa (Scapagnini et al., Mol. Neuribiol. 2011). The carotenoids, such as astaxanthin, on account of their extensive conjugation systems, are very efficient scavengers of singlet oxygen and, under low oxygen tension, of peroxy radicals.

Studies of intervention in humans with diets rich in carotenoids have demonstrated photoprotection of the skin, measured by decreased sensitivity to erythema induced by UV radiation. The capacity of some carotenoids for inducing phase 2 cytoprotective enzymes had previously been demonstrated. The applicant has demonstrated for the first time the capacity of astaxanthin for greatly improving the capacity of curcumin or of the related extract and for activating the Nrf2 pathway and, consequently, for causing expression of defence and repair genes, such as HO-1. Without wishing to demonstrate a specific molecular mechanism to explain this effect, the phenomenon might be due to the fact that dietary carotenoids and their metabolites share with all the other classes of phase 2 inducers, such as electrophilic polyphenols, a common chemical property, the ability to react with sulphydryl groups, which are the specific sensors of the activation of Nrf2.

Moreover, the applicant determined, using an experimental model, the capacity of the two compounds for activating mitochondrial biogenesis, a mechanism directly linked to cellular longevity and to the metabolic efficiency of the organism. Mitochondrial biogenesis is an elaborate cellular process that is based on the close linkage of various regulatory controls, from nuclear transcription of genes to the site-specific production of proteins. The pathways that regulate mitochondrial biogenesis have recently been considered as potential therapeutic targets for improving endothelial dysfunctions and vascular diseases observed in metabolic diseases.

The protective and anti-ageing action of the extract of Curcuma longa, of astaxanthin and of the mixture of the two active ingredients, from which the synergistic effect arises, was carried out by the following experiments. Activation of expression of Nrf2 in various cells

Three different cultures of cell lines, cortical neurons, skin fibroblasts and endothelial cells, were exposed to the extract of Curcuma longa, to astaxanthin or to a combination of the two compounds at a final concentration of 25 μΜ, to evaluate expression of the protein Nrf2 over time.

As shown in Fig. 1, treatment with the extract of Curcuma longa caused a significant time- dependent increase in expression of Nrf2 in nuclear extracts. Quantification of three independent Western blots showed that after one hour of exposure to 25 μΜ of extract of Curcuma longa, expression of Nrf2 increased significantly and remained upregulated for up to 12 hours, while the level of the transcription factor housekeeping Spl was stable. Astaxanthin induced a smaller increase in nuclear Nrf2. The combination of the two compounds massively induced expression of Nrf2 both when the concentration of curcumin and astaxanthin together was 25 μΜ (Fig. Id), and, even more clearly, when they were used respectively at a concentration of 25 μΜ, i.e. a total of 50 μΜ of active ingredients present (Fig. le).

Upregulation of HO-1 and of phase II detoxification enzymes in various cells

Exposure of three different cultures of cell lines, cortical neurons, skin fibroblasts and endothelial cells, for 6 hours to various concentrations of extract of Curcuma longa, astaxanthin or to a combination of the two compounds (5, 15, 25 and 50 μΜ) resulted in a significant increase (p < 0.05) of HO-1 mRNA, measured by real-time PCR, with a maximum value at 25 μΜ (Figs. 2a, b).

The extract of Curcuma longa causes a dose-dependent increase of HO-1 mRNA, measured relative to the non-inducible paralogous gene HO-2, reaching the maximum (about six times) at 25 μΜ, and then decreasing at 50 μΜ. Astaxanthin is undoubtedly much less active in induction of expression of the HO-1 gene at the same concentrations and reaches a peak again at 25 μΜ. To confirm that expression of the HO-1 gene measured at mRNA level corresponded to an equivalent increase of activity of haem oxygenase, this activity was measured at the same doses of extract of Curcuma longa and of astaxanthin, 6 and 24 hours after administration. The course of the increase in activity was comparable to the results obtained when examining cellular mRNAs, confirming the functional significance of the data obtained using determinations with real-time PCR (Figs. 2c, d, e). It has thus been demonstrated that the simultaneous administration of curcumin and astaxanthin allows an increase in activity of haem oxygenase to be reached that cannot be compared with that obtained with the two substances individually. This result demonstrates the capacity of the compounds for stimulating the cells of the body, in particular neurons, skin fibroblasts and endothelial cells, to repair the damage due to the ageing process.

Neuroprotective effects of the invention The field selected for experiments to test the capacity of the compounds for improving cell survival was a model of cell death induced in rat cortical neurons exposed to oxidative stress.

Neurons are highly sensitive to pro-oxidant stimulation and the biological effects at the focus of this experiment were the capacity of the compounds for promoting positive modulation of neuronal survival and positive modulation of neuronal repair and plasticity, both in terms of single neurons, and of the neuronal population.

The applicant therefore decided to use the combination of the two compounds, extract of Curcuma longa and astaxanthin, both at 25 μΜ, before exposing the cells for 2 hours to glucose oxidase (50 mU/ml). The oxidant system of glucose oxidase generates hydrogen peroxide at a constant rate and is known to produce cellular adhesion in vitro.

After treatment with glucose oxidase, the cells were washed and exposed to complete medium containing 1% of Alamar blue for 5 hours, according to the manufacturer's instructions, for determining cellular vitality. After the incubation time, the optical density of the medium in each well was measured using a plate reader. The assay is based on detection of the metabolic activity of living cells, using a redox indicator that changes its form from oxidized (blue) to reduced (red). The intensity of the red colour is proportional to the vitality of the cells.

Treatment of cells with glucose oxidase for 2 hours resulted in 27% of residual cellular vitality (Fig. 3). Exposure of cells for 12 hours to the extract of Curcuma longa 25 μΜ reduced the damage caused by glucose oxidase, increasing cellular vitality by 55% (28% more than the control).

Astaxanthin at a concentration of 25 μΜ was less effective in protecting cells from oxidative damage, giving a vitality of 43% (15% greater than the control). The combination of the two compounds conferred protection on the cells synergistically, giving a degree of neuronal survival of 91% (64% greater than the control, which is greater of 43% than the expected additive effect). The combination of the two compounds at the same concentration as when used individually (both at 25 μΜ) further increases the protective effect, even succeeding in stimulating proliferation despite the oxidative damage exerted. In fact, counting shows an increase in the number of neuronal cells having a degree of survival at 109%.

Induction of mitochondrial biogenesis

As widely demonstrated in the literature and explained well by G. Lopez-Lluch et al., in "Mitochondrial biogenesis and healthy aging" (Exp Gerontol. 2008 September; 43(9): 813— 819), decrease in the production of mitochondria by the cells is a process linked to ageing of the cell and its progressive decline of functionality. Maintaining and/or stimulating mitochondrial genesis in cells means prompting them to remain efficient and to prolong their vitality.

The cells were treated with extract of Curcuma longa (25 μιηοΐ/ΐ for 48 hours), astaxanthin (25 μηιοΐ/ΐ for 48) or with extract of Curcuma longa and astaxanthin. The mitochondrial mass in the endothelial cells was determined by loading the mitochondria selectively with the red fluorescent dye Mitotracker red (Invitrogen, Carlsbad, CA). The fluorescent dye calcein (green) and Hoechst 33258 (blue) were used for staining the cytoplasm and the nucleus, respectively. Optical sections of HUVAs were obtained at 60% magnification and the ratio of the densities of mitochondrial area relative to the cytoplasm volume was calculated using the Zeiss Axiovision visualization software.

Only cells with intact calcein staining of the cytoplasm were included in the analysis. The total DNA was isolated from the endothelial cells (DirectPCR; Viagen Biotech). The copy number of the mitochondrial DNA (mtDNA) was determined by QRT-PCR as already described, using cytochrome oxidase III and β-actin as markers for the copy numbers of mtDNA and of genomic DNA, respectively.

The effect of treatment with extract of Curcuma longa, astaxanthin or extract of Curcuma longa and astaxanthin together, at two concentrations, firstly at 25 μΜ together and then at 50 μΜ in total, given by the sum of 25 μΜ of Curcuma longa and 25 μΜ of astaxanthin, was determined.

Staining with Mitotracker showed that the mitochondria are localized in the perinuclear region in endothelial cells (Fig. 4). The ratio of density of mitochondrial area to cytoplasm volume in endothelial cells labelled with Mitotracker was increased significantly by the treatment with extract of Curcuma longa (Fig. 4).

Analysis of the intensities of fluorescence of Mitotracker in EC confirmed that the treatment with extract of Curcuma longa increases the mitochondrial mass significantly in the treated cells compared with the untreated cells.

Astaxanthin was not effective in terms of activation of the mitochondrial mass, but the extract of Curcuma longa in combination with astaxanthin was significantly more effective compared with the extract of Curcuma longa alone (Fig. 4). The formulation of the invention was also able to improve the levels of mtDNA considerably, as shown in Fig. 4.

Mitochondrial biogenesis involves the integration of a large number of transcriptional pathways that control nuclear mitochondrial gene expression. The coactivator PPAR- PGC-1 a, Nrfl and Tfam are considered to be key regulators of mitochondrial biogenesis in many tissues. The applicant used a QRT-PCR technique for determining the effect of the extract of Curcuma longa, astaxanthin and extract of Curcuma longa in combination with astaxanthin (25 μιηοΐ/ΐ for 24 hours) on the mRNA expression of the mitochondrial biogenesis factors nuclear respiratory factor- 1 (Nrfl), mitochondrial transmission factor A (Tfam) and peroxisome proliferator-activated receptor (PPAR)-a coactivator- 1 (PGC-1 a) in endothelial cells using Strategen MX3000 as reported.

Total RNA was isolated with a mini RNA Isolation Kit (Zymo Research, Orange, CA) and was reverse-transcribed using Superscript III RT (Invitrogen) as previously described. The efficiency of the PCR reaction was determined using dilution series of a standard sample. Quantification was performed using an efficiency-regulated CT method. The housekeeping gene HPRT was used for internal normalization.

The fidelity of the PCR reaction was determined by melting point analysis and visualization of the product in 2% agarose gel. The QRT-PCR measurements revealed that expression of the mitochondrial biogenesis factors PGC-1 a, Nrfl and Tfam in endothelial cells (Fig. 5) was significantly increased in the treatment with the formulation according to the invention.

Protective and regenerative effects of the invention on human fibroblasts

UV radiation induces production of metalloproteases (MMP) that destroy collagen I and III, and decreases that of type I procollagen in the dermis. Taken together, these events lead to loss of tonicity and firmness of the skin typical of the ageing process. The applicant analysed the effect of the extract of Curcuma longa (25 μΜ), of astaxanthin (25μΜ), or of the combination of the two (25μΜ and 50 μΜ) in protecting human fibroblasts from photo-ageing induced by UV radiation. Fibroblasts of the human line Hs68 were pretreated for 24h with the test substances. Then the culture medium was replaced with PBS and the cells were exposed to UV light (20 mJ/cm2). A quantitative analysis was performed by real-time PCR of the mRNA of MMP- 1 and of type I procollagen.

The treatment with extract of Curcuma longa and with astaxanthin decreases the levels of mRNA of MMP-1 induced by UV radiation significantly, but a greater reduction is obtained when the two substances are used together and have a synergistic effect. Activity of stimulation of expression of mRNA of procollagen I is explained similarly (Fig. 6).

The formulation of the present invention can moreover contain other ingredients, as in the examples that follow. Based on preliminary studies demonstrating the synergistic effect between the extract of Curcuma longa and astaxanthin, the applicant conducted extensive research on cultured rat endothelial and neuronal cells, exposing them to different concentrations (10, 25, 50 μΜ/ml) of the aforementioned formulation according to the invention, for 6, 12 and 24 hours.

The study demonstrated the capacity of the formulation according to the invention for strongly inducing the expression of haem oxygenase- 1 (HO-1), a highly protective protein in defence against oxidative stress (Poon HF, Calabrese V, Scapagnini G, Butterfield DA (2004) J. Gerontol., A Biol. Sci., Med. Sci. 59(5): 478-493), in treated cells and therefore in protecting the cells from damage caused by exposure to free radicals.

Haem oxygenase is the principal enzyme for catabolism of haem, which is broken down by biliverdin-bilirubin, carbon monoxide (CO) and iron. These products are molecules having particularly significant biological activity: biliverdin and bilirubin have potent antioxidant activities (Scapagnini G et al. (2002) Brain Res. 654(1): 51-59); CO performs fundamental signalling functions and behaves as a gaseous modulator in vascular function (Abraham NG, Scapagnini G, Kapas A (2003) J Cell. Biochem. 90(6): 1098-1 1 11); iron is a known gene regulator (Smith MA et al. (1998) Brain Res. 788, 232-236).

In addition to generating molecules that are active against oxidative stress, HO-1 lowers the levels of haem, which is intrinsically a potent inducer of lipid peroxidation and formation of free radicals. Its activation appears to represent an important defence mechanism for neurons exposed to oxidative damage in acute situations, such as infarction, or chronic conditions, such as cerebral ageing, or during neurodegenerative diseases, such as Alzheimer's disease.

Undoubtedly, both oxidative damage and inflammatory processes are particularly elevated in the brain of patients with Alzheimer's disease, suggesting a potential benefit for long- term treatment with a compound undoubtedly endowed with potent antioxidant and anti- inflammatory action. Research has also shown that induction of HO-1 by the formulation of the invention is correlated with capacity for activating Nrf2. This transcription factor is capable of strongly inducing the expression of other protective genes, such as detoxifying enzymes of type 2, which have high antioxidant and antitumour activity. Moreover, in the same experimental conditions, the formulation according to the invention is capable of activating mitochondrial biogenesis considerably. The experimental data support the fact that the formulation increases the mitochondrial content in endothelial cells (Fig. 4a). Increased mitochondrial biogenesis in cells treated with the formulation is also indicated by the increased content of cellular mtDNA (Fig. 4b). Multiple mechanisms can explain the mitochondrial biogenesis induced by the formulation and its contribution to cellular health. To determine whether the increased number of mitochondria in endothelial cells treated with the formulation is the result of induction of mitochondrial biogenesis factors, QRT- PCR was used for examining the expression of Nrfl, Tfam and PGC-1 a. Nrfl activates the transcription of numerous components encoded by the nucleus of the electron-transport chain and also regulates Tfam, which is responsible for the transcription of encoded mtDNA genes.

The regulatory function of Nrfl and of other mitochondrial biogenesis factors is modulated by PGC-1 a. It has been demonstrated that the formulation according to the invention induces Nrfl , Tfam and PGC-1 a (Fig. 5).

It has been demonstrated that the altered expression of these factors modulates the activity of mitochondrial biogenesis and improves cellular longevity.

The compositions of the present invention are prepared in dosage forms suitable for concomitant administration of various components within the following dosages expressed as mg/day:

a) extract of Curcuma longa from 10 to 12 000

b) astaxanthin from 0.25 to 25.

According to a preferred aspect, the dosage forms of the present invention will comprise the various components within the following dosages expressed as mg/day:

a) extract of Curcuma longa: from 60 to 1100;

b) astaxanthin: from 0.5 to 4.

The dosage forms can comprise amounts of the aforementioned components a) and b) that are submultiples from 2 to 5 of the aforementioned amounts. Commercial extracts of Curcuma longa can have a content of curcuminoids up to 95 wt%, for which the amounts and the dosages stated above relating to the extract are likewise valid in relation to amounts and dosages of curcuminoids up to 95% of that stated.

A suitable formulation, particularly as a dietary supplement, advantageously contains the following ingredients in the following unit doses:

extract of Curcuma longa or content of curcuminoids from 50 to 600 mg,

astaxanthin from 0.5 to 4 mg.

According to a preferred aspect of the invention the extract of Curcuma longa is present as a complex with a soy phospholipid, such as lecithins, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and the like, in particular soy distearoylphosphatidylcholine. These complexes, known as phytosomes, are described for example in EP 0 441 279 and are available commercially under the trade name Fitosoma®. The lipophilic portion of this complex, soy phospholipid, permits better interaction of the active ingredient with the cholic acids responsible for emulsifying and capturing the substances to be transported, for absorption, to the systemic circulation. Toxicological studies conducted on animals and pharmacokinetic studies in humans have in fact demonstrated absence of toxicity and greater absorption of the ingredient administered as phytosome compared with the ingredient administered in free form.

According to a preferred aspect of the present invention, the compositions in question can contain additional components, with therapeutic action, or complementary, or otherwise useful for the purposes of the invention. Examples of said additional components are plant extracts, vitamins, nutrients, trace elements, and the like.

These additional components will be present in amounts corresponding to their recommended daily dose (or to submultiples or multiples thereof). Examples of said additional components are extracts of black pepper, rosemary, Vitis vinifera, Boswellia serrata, Melissa officinalis, Ginkgo biloba; nutrients such as fish or yeast, nucleotides, resveratrol, lycopene; group B vitamins, vitamin D, vitamin C, vitamin K and vitamin E; essential oils such as D-limonene and trace elements such as Mg, Mn, K in the various salifications with inorganic and organic salts and in aminochelate forms.

The compositions of the invention can be formulated in any suitable form for oral administration, for example as hard or soft gelatin capsules, tablets, effervescent or chewable tablets, granules or powders in sachets, solid forms with controlled release, chewing gums and the like.

The compositions of the present invention can be formulated in a suitable way for administration by the oral route and will be prepared according to conventional methods that are well known in pharmaceutical technology, such as those described in "Remington's Pharmaceutical Handbook", Mack Publishing Co., N.Y., USA, using excipients, diluents, fillers or anti-agglomerating agents acceptable for pharmaceutical, cosmetic or food use. FORMULATION EXAMPLES (dose/unit or treatment)

Example 1 - TABLETS 1

Extract of Curcuma longa complexed with phospholipids 350 mg Astaxanthin 2 mg

Microcel® 40 mg

Magnesium stearate 5 mg

Aerosil® 10 mg Example 2 - FILM-COATED TABLETS

Extract of Curcuma longa complexed with phospholipids 350 mg

Astaxanthin 2 mg

Microcel® 340 mg

Dicaphos® 333 mg

Magnesium stearate 10 mg

Aerosil® 8 mg

Explocel® 18 mg

Sepifilm® 22.8 mg

Shellac 7 mg

Colorant 0.2 mg

Example 3 - FILM-COATED TABLETS Extract of Curcuma longa complexed with phospholipids 4 0 mg Astaxanthin 1 mg

Extract of Melissa officinalis 60 mg

Extract of Boswellia serrata 100 mg

Extract of rhatany 5 mg

Vitamin D3

Microcel® 340 mg

Dicaphos® 333 mg Magnesium stearate 10 mg

Aerosil® 8 mg

Explocel® 18 mg

Sepifilm® 22.8 mg

Shellac 7 mg

Colorant 0.2 mg

Example 5 - SACHETS Extract of Curcuma longa complexed with phospholipids 200 mg Astaxanthin 4 mg

Limonene microencapsulated by coacervation 75 mg

Extract of rosemary 10 mg

Fructose 1873 mg

Aerosil® 15 mg

Orange flavour 180 mg

Citric acid 220 mg

Acesulfame K 25 mg

E102 2 mg

Example 6 - DISPERSIBLE ORAL FORMULATION

Extract of Curcuma longa complexed with phospholipids 50 mg Astaxanthin 1 mg

Vitamin E

Limonene o.e. 5 mg

Sorbitol 160 mg

Orange flavour 20 mg

Mandarin flavour 5 mg

Acesulfame K 2 mg

Aerosil® 5 mg

Fructose 1530 mg Example 7 - SOFT GELATIN CAPSULES

Extract of Curcuma longa complexed with phospholipids 350 mg Astaxanthin 0.5 mg

Lycopene 0.3 mg

Piperine 2.5 mg

Extract of rosemary 10 mg

Resveratrol 5.50 mg

Nucleotides 3.50 mg

Limonene o.e. 130 mg

Vitamin E 6 mg

Vitamin D3 2.50 mg

Food-grade oil q.s.

Claims

1. Pharmaceutical or dietary supplement composition, comprising, as active ingredients with synergistic or combined effect, curcumin and astaxanthin, for use for oral administration in the prevention and treatment of neurodegenerative disorders of the central and/or peripheral nervous system and of diseases associated with ageing of the skin.

2. Composition according to Claim 1 , comprising extract of Curcuma longa as a source of curcumin.

3. Composition according to Claim 1 or 2, comprising, as a source of curcumin, an extract of Curcuma longa in the form of a complex with a soy phospholipid.

4. Composition according to Claim 3, wherein said soy phospholipid is selected from the group consisting of soy lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and distearoylphosphatidylcholine.

5. Composition according to any one of Claims 1 to 4, comprising, as a source of astaxanthin, an extract of Haematococcus pluvialis.

6. Composition according to any one of the preceding claims, in dosage form, comprising from 0.5 mg to 4 mg of astaxanthin and 50 mg to 600 mg of an extract of Curcuma longa or an extract of Curcuma longa complexed with a soy phospholipid.

7. Composition according to any one of the preceding claims, for use in oral administration, where said use comprises the combined, concomitant or sequential administration of an extract of Curcuma longa in amounts from 10 mg to 12 000 mg/day and of astaxanthin in amounts from 0.25 mg to 25 mg/day.

8. Composition according to Claim 7, for the combined, concomitant or sequential administration of an extract of Curcuma longa in amounts from 60 mg to 1 100 mg/day and of astaxanthin in amounts from 0.5 mg to 4 mg/.

9. Composition according to any one of the preceding claims, further comprising one or more extracts selected from the group consisting of extract of black pepper, of rosemary, of Vitis vinifera, of Boswellia serrata, of Melissa officinalis, and/or Ginkgo biloba.

10. Composition according to Claim 9, further comprising nutrients selected from the group consisting of fish or yeast nucleotides, resveratrol, lycopene, group B vitamins, vitamin D, vitamin C, vitamin K and vitamin E.

11. Composition according to any one of the preceding claims, further comprising essential oils and/or trace elements.