Mixture, compositions containing said mixture and their use in the treatment and / or prevention of neurodegenerative diseases affecting motor neurons and / or due to demyelination processes

Abstract

The present invention relates to a mixture, compositions comprising said mixture, and their use in the treatment and / or prevention of neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS). In greater detail, the present invention relates to a mixture comprising or, alternatively, consisting of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) lipoic acid, or a salt thereof, and / or a bergamot extract.

Description

[0001] MIXTURE, COMPOSITIONS CONTAINING SAID MIXTURE AND THEIR USE IN THE TREATMENT AND / OR PREVENTION OF NEURODEGENERATIVE DISEASES AFFECTING MOTOR NEURONS AND / OR DUE TO DEMYELINATION PROCESSES

[0002] ***

[0003] DESCRIPTION

[0004] The present invention relates to a mixture, compositions comprising said mixture, and their use in the treatment and / or prevention of neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0005] In greater detail, the present invention relates to a mixture comprising or, alternatively, consisting of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) lipoic acid, or a salt thereof, and / or a bergamot extract.

[0006] BACKGROUND OF THE INVENTION

[0007] Amyotrophic Lateral Sclerosis (ALS) is a degenerative disease that affects motor neurons, i.e. the neurons responsible for the contraction of the voluntary muscles involved in movement and in vital functions such as swallowing, breathing and speech. Neurodegeneration results in the loss of neuronal functions, axon deterioration and replacement of the neuronal part with the glial one. The muscles, no longer correctly innervated, also undergo atrophy and the formation of sclerotic plaques.

[0008] The etiopathogenesis is not yet well defined, but there are several known markers and conditions, present both in sporadic forms, and in familial ones, which have been recognised as underlying the onset of this disease. These include the SOD1 gene mutation, which triggers the synthesis of an aberrant protein that accumulates in toxic aggregates; an excess of glutamate, which is harmful because it causes hyperexcitation and an increase in calcium in the synaptic space; a high rate of neuroinflammation, characterised by astrogliosis, microglia activation and infiltration of peripheral immune cells at the site of neurodegeneration; and an excessive production of reactive oxygen species, due to the inflammation, which contribute to creating an environment that is toxic for cell life.

[0009] Clinically, the disease manifests itself through limb weakness, cramps, frequent falls, and a generalised state of spasticity and hyperreflexia. In more advanced stages, dysarthria and dysphagia are also observed, with a high risk of aspiration pneumonia and an excessive production of saliva and difficulty in swallowing it (sialorrhea). In 50% of patients, one also witnesses the appearance of the pseudobulbar affect, i.e. outbursts of unmotivated laughing or crying not consistent with their psychophysical state, or frequent yawning, a clear signal of neuronal degeneration. Ten percent of patients also develop dementia, caused by the degeneration of frontotemporal neurons. The currently available therapies have the aim of reducing the release of glutamate and reducing oxidative stress. To date, the only drug approved for the treatment of ALS is Riluzole, which has shown effectiveness in prolonging the median survival of patients by three months compared to the placebo. This drug acts by counteracting the release of glutamate; therefore, its effectiveness is less significant in advanced stages of the disease, so it is important that it be administered as soon as possible following the diagnosis. In combination with Riluzole, symptomatic treatments are applied; these include the use of strong anticholinergic drugs to reduce sialorrhea, tricyclic antidepressants or selective serotonin reuptake inhibitors for the pseudobulbar affect, centrally acting muscle relaxants and specific physiotherapy for the cramps and spasticity. Despite this, the prognosis is poor, and life expectancy stands at around 3-5 years after the time of diagnosis.

[0010] The term Multiple Sclerosis (MS), relates to the more common disabling disease of non-traumatic origin. The causes are still unknown; it is believed that different pathways are involved and that the main pathological mechanisms imply neuroinflammation, demyelination and axon degeneration processes. The most accredited theory is that MS starts off as an inflammatory immune-mediated disorder, distinguished by autoreactive lymphocytes having myelin as their main target and which, through their activity, cause demyelination and axonal damage. For this reason, the disease is considered to be of the autoimmune type. Subsequently, the progression of the disease is characterised by microglia activation processes and chronic neurodegeneration, with an accumulation of macrophages in the damaged tissues. In addition to the loss of myelin, another typical pathological feature is the presence of sclerotised plaques in different brain locations, accompanied by neuroinflammation and gliosis. The progression of the disease is characterised by a degenerative phase of brain atrophy and axonal loss. The activated microglia damage oligodendrocytes by secreting pro-inflammatory cytokines and neurons through the production of ROS and RNS, thus inducing neuronal dysfunction.

[0011] Although there are no unique clinical signs for MS, there do exist some that characterise the disease, which can present as monosymptomatic, if there is a single lesion, or polysymptomatic, if multiple lesions affect several areas of the nervous system. Patients affected by MS are typically young adults aged between 20 and 40, who report debilitating tiredness and fatigue not correlated with physical activity. A sensation of pain on different levels is also very common: headache, neuropathic pain, back pain and painful spasms. Also reported are sleep disorders of varying entity, motor symptoms such as paraparesis and paraplegia, dysphagia, sensory symptoms, sensitivity to heat (Uhthoff's Syndrome), spasticity, paroxysmal symptoms, visual symptoms, dizziness, eye movement abnormalities, epilepsy, depression - observed in over two thirds of patients - and cognitive deficits, associated with problems tied above all to attention, executive function, abstract conceptualisation and memory. Finally, about 50% of patients suffer from intestinal dysfunctions, including constipation, limited bowel movements and incontinence; whereas 75% are affected by bladder dysfunctions, which cause disorders tied mainly to urinary urgency and bladder filling incapacity and urine retention. The progression of the disease can be characterised by flare-ups alternating with periods of clinical stability (relapsing-remitting phenotype), until it transitions into a stage in which the neurodegeneration is progressive and constant (secondary progressive phenotype), or else it starts immediately with a constant, progressive character (primary progressive phenotype).

[0012] As regards treatments for MS, these can be targeted at flare-ups or symptoms, or at modifying the disease. The most widely used therapy to combat relapses is the administration of steroids, but this is not always effective or well tolerated. In recent years, a breakthrough in the treatment of MS has been made possible thanks to the advent of disease modifying therapies. These therapeutic strategies mainly provide for the use of immunosuppressive and immunomodulatory drugs and therapies aimed at reconstructing and reinforcing the immune system; however, none of the treatments designed to modify MS shows to provide a definitive solution. In fact, their use is aimed solely at preventing relapses and reducing the accumulation of inflammation, brain dysfunctions and, consequently, neurological disability. Finally, symptomatic therapies are used to treat the vast array of symptoms typical of MS and for the most part they are pharmacological and physical therapies, but not MS-specific ones, and they have no action with respect to neuronal damage. Such therapies are mainly aimed against fatigue, mood disorders, neuropathic pain, bowel and urinary disorders, and motor disorders.

[0013] Considering the physiopathology of both diseases described, ALS and MS, the importance of intervening in neuroprotection in the early stages of the onset of symptoms, pending an actual diagnosis, is clearly evident and, consequently, it is necessary to develop a nutraceutical support, free of side effects, which targets the causes of neurodegeneration in order to prevent it or slow it down.

[0014] The technical problem that the present invention addresses and solves is thus to provide a valid and effective solution for maintaining the health of the central nervous system, preferably to counteract motor neuron degeneration and / or demyelination processes, in the absence of side effects and / or adverse reactions, and which is useful for treating Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0015] Furthermore, it is necessary to provide a mixture and / or a composition which are useful for that purpose, easy to prepare and free of side effects, and which do not require the use of costly machinery and processes.

[0016] These aims, and still others which will emerge clearly from the detailed description that follows, are achieved by the mixtures and the compositions comprising said mixtures (mixtures and / or compositions of the invention) which have the technical features claimed in the appended claims.

[0017] SUMMARY OF THE INVENTION

[0018] Following an intense research activity, the Applicant has developed and prepared a mixture comprising or, alternatively, consisting of specific active components which are all natural ingredients and thus make both the mixture and the pharmaceutical composition or nutraceutical composition or supplement composition comprising said mixture free of side effects and characterised by high tolerability.

[0019] In fact, the mixture according to the invention or the composition comprising it has no significant adverse effects. Precisely for these reasons, it can be advantageously administered to a broad category of individuals, including immunocompromised individuals.

[0020] A first aspect of the present invention relates to a mixture comprising or, alternatively, consisting of: i) an extract of chamomile; ii) an extract of turmeric; ill) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) lipoic acid, or a salt thereof, and / or a bergamot extract.

[0021] Preferably, the mixture according to the invention comprises v) a bergamot extract.

[0022] Therefore, more preferably, the mixture according to the invention comprises or, alternatively, consists of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) a bergamot extract.

[0023] In the context of the present invention, in the experimental section which follows and in the appended figures, the term "Mix 1” is meant to indicate the mixture i) +ii) + iii) + iv) as defined above and v) lipoic acid or a salt thereof.

[0024] In the context of the present invention, in the experimental section which follows and in the appended figures, the term "Mix 2” is meant to indicate the mixture i) +ii) + iii) + iv) as defined above and v) a bergamot extract.

[0025] A second aspect of the present invention relates to a composition comprising said mixture, according to said first aspect of the invention, and food or pharmaceutical grade excipients and / or additives.

[0026] Said composition is, for example, a pharmaceutical composition or a nutraceutical composition or a supplement composition or a food composition or a composition for medical devices, for example according to Reg. (EU) 745 / 2017, or a composition for foods for special medical purposes, for example according to Reg. (EU) 609 / 2013 (in short, the compositions of the invention).

[0027] Preferably, the composition of the invention may also comprise at least one further ingredient selected from the group comprising or, alternatively, consisting of at least one vitamin selected in the group comprising or, alternatively, consisting of vitamin C, vitamin E, vitamin B1 , vitamin B2, vitamin B6 and vitamin B12 or mixtures thereof; L-tryptophan; a strawberry extract, for example an extract of strawberry Fragaria,' an extract of Crocus sativus, or mixtures thereof.

[0028] A third aspect of the present invention relates to the mixture Mix 1 and Mix 2 according to the invention and / or the composition comprising said mixture for use as a medicament.

[0029] The mixture of the invention and the pharmaceutical and / or nutraceutical composition comprising said mixture are particularly useful for maintaining the wellbeing and health of the central nervous system, preferably to prevent and / or treat neurodegenerative diseases affecting motor neurons and / or due to demyelination processes.

[0030] Therefore, the mixture of the invention and / or the composition comprising said mixture can be advantageously used in a method for protecting central nervous system function and / or for use in a method for counteracting motor neuron degeneration and / or demyelination processes and / or for use in a method for preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0031] DESCRIPTION OF THE FIGURES

[0032] Figure 1 : cell viability following treatment with the single substances and mixture. All concentrations p<0.05 vs the control; #p<0.05 vs single substances; y p<0.05 vs Riluzole.

[0033] Figure 2: cell viability following pretreatment with glutamate toxic stimulation and treatment with the single substances and mixture. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate.

[0034] Figure 3: ROS production following toxic stimulation with glutamate and treatment with the single substances and mixture. *p<0.05 vs the control; #p<0.05 vs the single agents; <p p<0.05 vs glutamate.

[0035] Figure 4: % inhibition of nitric oxide (NO) production induced by toxic stimulation with glutamate and treatment with the single substances and mixture. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate. Figure 5: glutathione (GSH) production levels in a model of neurodegeneration caused by glutamate excitotoxicity. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate.

[0036] Figure 6: expression of markers of neurodegeneration caused by glutamate-induced damage: A) SOD and B) ADAR2. * p<0.05 vs the control; # p<0.05 vs the single compounds; <p p<0.05 vs glutamate; bar p<0.05 vs Mix 1 .

[0037] Figure 7: cell viability under conditions of A) pretreatment, B) co-treatment and 0) post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate; ip p<0.05 vs Riluzole.

[0038] Figure 8: analysis of the condition of oxidative stress under the three experimental conditions of A) pretreatment, B) co-treatment and 0) post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate; ip p<0.05 vs Riluzole.

[0039] Figure 9: analysis of the condition of nitrosative stress under the three experimental conditions of A) pretreatment, B) co-treatment and C) post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate; ip p<0.05 vs Riluzole.

[0040] Figure 10: analysis of lipid (10 A-C) and protein (10 D-F) peroxidation under the three experimental conditions of pretreatment, co-treatment and post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; cp p<0.05 vs glutamate; ip p<0.05 vs Riluzole.

[0041] Figure 11 : quantification of intracellular calcium under the three experimental conditions of A) pretreatment, B) cotreatment and C) post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate; ip p<0.05 vs Riluzole.

[0042] Figure 12: analysis of the neurodegeneration biomarkers SOD (12 A-C) and ADAR2 (12 D-F), altered in the event of motor neuron degeneration and axon demyelination, under the three experimental conditions of pretreatment, co-treatment and post-treatment with Riluzole. * p<0.05 vs the control; # p<0.05 vs the single agents; <p p<0.05 vs glutamate; ip p<0.05 vs Riluzole. Figure 13: passage through the blood-brain barrier (BBB in the Figure) of the single components indicated in the figure and of the mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0043] Figure 14: analysis of the two tight junction markers A) MarvelD3 and B) claudin-5, safety of the passage of the single components and of Mix 1 and 2 through the blood-brain barrier according to what is described in example 6. Figure 15: analysis of cell viability after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0044] Figure 16: ROS Production following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0045] Figure 17: NO Production following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0046] Figure 18: GSH quantification following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0047] Figure 19: Glutamate release following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0048] Figure 20: Calcium release following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0049] Figure 21 : SOD activity following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0050] Figure 22: TDP43 activity following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0051] Figure 23: ADAR2 activity following toxic stimulation with glutamate and treatment with the single substances and mixtures Mix 1 and Mix 2 according to what is described in example 6.

[0052] Figure 24: analysis of cell viability after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) co-treatment and C) posttreatment with Riluzole according to what is described in example 6.

[0053] Figure 25: analysis of lipid peroxidation after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) co-treatment and C) post-treatment with Riluzole according to what is described in example 6.

[0054] Figure 26: analysis of protein peroxidation after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) cotreatment and C) post-treatment with Riluzole according to what is described in example 6.

[0055] Figure 27: glutamate release after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) co-treatment and C) posttreatment with Riluzole according to what is described in example 6. Figure 28: TDP43 levels after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) co-treatment and C) post-treatment with Riluzole according to what is described in example 6.

[0056] Figure 29: ADAR2 levels after glutamate-induced damage and treatment with the single substances and mixtures Mix 1 and Mix 2 under the three experimental conditions of A) pretreatment, B) co-treatment and C) post-treatment with Riluzole according to what is described in example 6.

[0057] DETAILED DESCRIPTION OF THE INVENTION

[0058] The first aspect of the present invention relates to a mixture (Mix 1 and Mix 2) comprising or, alternatively, consisting of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) lipoic acid, or a salt thereof, and / or a bergamot extract.

[0059] The mixture of the present invention Mix 1 and Mix 2 comprises I) an extract of chamomile.

[0060] In the context of the present invention, extract of chamomile is understood to include all types of extracts of common chamomile comprising the flavone apigenin, such as, for example, Matricaria chamomile and Matricaria recutita L. Preferably, I) said extract of chamomile is a dry extract and is for example a dry extract from the flowers of Matricaria chamomile, such as, for example, a dry extract titrated in apigenin, the minimum apigenin titre being > 2% w / w, the apigenin titre preferably being in a range of from 5% to 10% by weight relative to the total weight of the dry extract. The mixture of the invention may for example comprise a dry extract of chamomile, preferably extracted from the flowers of Matricaria recutita L, for example with ethanol and water.

[0061] Matricaria chamomile and Matricaria recutita L. are two names for the same plant, commonly known as common chamomile or German chamomile. The name "Matricaria recutita L." is the correct scientific name according to the botanical classification system, whereas "Matricaria chamomile" is a more dated synonym, but still widely used. In the context of the present invention, the term "Matricaria chamomile" indicates the plant Matricaria recutita L.

[0062] In order to prepare a dry extract, for example a dry extract of chamomile, one generally uses extraction with a solvent followed by purification and concentration of the product. Titration of the active compound present in the extract is then carried out by chromatography techniques such as gas chromatography (GC) or high-performance chromatography (HPLC) to determine the concentration of the desired compound.

[0063] Main preparation steps:

[0064] 1. Extraction:

[0065] Flowers of Matricaria recutita L are ground and subjected to extraction with an organic solvent (e.g. ethanol). This process allows for solubilising the liposoluble compounds. 2. Purification:

[0066] After extraction, the solvent is removed, leaving a raw extract containing a mixture of compounds. This extract can be purified by means of techniques such as molecular distillation or column chromatography to remove unwanted components and concentrate the compound of interest, for example apigenin.

[0067] 3. Titration:

[0068] The purified extract is analysed by means of chromatography techniques (GC or HPLC) to determine its concentration. These techniques separate the components of the extract based on their chemical properties and allow for their quantification.

[0069] 4. Formulation:

[0070] The titrated purified extract can then be formulated into different products, such as food supplements.

[0071] One example of a commercial product is the dry extract of chamomile flowers (botanical name: Matricaria recutita L.) sold by Vivatis Pharma, which has a minimum apigenin titre of 5% as measured by HPLC and appears as a powder having a min. particle size of 95% passing through an 80 mesh sieve, and a bulk density comprised from 0.3 g / ml to 0.6 g / ml, measured as per CP <2020>.

[0072] Preferably, said extract shows a loss on drying of max. 5%, measured at 5 g / 105°C / 2hrs, and an ash content of max. 5%, measured at 2 g / 525°C / 3hrs.

[0073] The mixture of the present invention, Mix 1 and Mix 2, comprises, in addition to I) an extract of chamomile, also ii) an extract of turmeric.

[0074] For example, the mixture of the invention may comprise a dry extract of turmeric.

[0075] In the context of the present invention, extract of turmeric is understood to include all types of turmeric root extracts comprising curcuminoids. Preferably, the extract of turmeric is a root extract from the plant Curcuma longa.

[0076] Preferably, said extract of turmeric is a dry extract titrated in curcuminoids, the minimum curcuminoid titre being > 30% w / w as measured by HPLC, the curcuminoid titre preferably being in a range of from 30% to 50%, preferably comprised from 35% to 45%, for example 35% by weight relative to the total weight of the dry extract. For example, the extract of turmeric present in the mixture according to the invention can be the product CurGfen® Powder NF, for example sold by Akay Flavours & Aromatics Pvt. Ltd. Malaidamthuruthu P. O. This product contains an extract of turmeric obtained from the rhizome and seeds. The product appears as a yellow / orange-coloured powder, with a particle size of 90% passing through a 60 mesh sieve and a bulk density comprised from 0.3 to 0.7 g / ml, measured as per USP 29 <616>.

[0077] CurGfen® Powder NF has a curcuminoid titre (curcumin, DMC, BDMC) of 35% as measured by HPLC.

[0078] The mixture of the invention, Mix 1 and Mix 2, comprises, in addition to I) an extract of chamomile and ii) an extract of turmeric, also ill) L-acetyl carnitine.

[0079] L-acetyl carnitine, IUPAC name (3R)-3-acetyloxy-4-(trimethylazaniumyl)butanoate hydrochloride (CAS: 3040-38- 8), also known as LAC, acetyl-L-carnitine, levacecarnine or ALCAR, is an acetylated form of L-carnitine or carnitine. In the context of the present invention the terms "L-acetyl carnitine” or “acetyl-L-carnitine” are meant to refer to, and include, in general, L-acetyl carnitine as such and / or in the form of a salt thereof and / or mixtures thereof, and / or L- carnitine and / or carnitine.

[0080] Preferably, the mixture according to the invention comprises L-acetyl carnitine in the form of a salt, preferably L- acetyl carnitine hydrochloride. For example, the mixture may comprise i) an extract of chamomile, as described above, ii) an extract of turmeric, as described above, and iii) L-acetyl carnitine in the form of a salt, preferably L- acetyl carnitine hydrochloride.

[0081] For example, the L-acetyl carnitine present in the mixture is the commercial product having the name ACETYL-L- CARNITINE HCI MATRIS®, sold by the firm IPS.

[0082] The mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i) an extract of chamomile, ii) an extract of turmeric, iii) L-acetyl carnitine, preferably L-acetyl carnitine in the form of a salt, preferably L-acetyl carnitine hydrochloride, also iv) a blueberry extract.

[0083] In the context of the present invention, extract of Vaccinium myrtillus is understood to include all types of blueberry extracts comprising anthocyanins. Preferably, the blueberry extract {Vaccinium myrtillus) is a dry extract titrated in anthocyanins, the minimum anthocyanin titre being > 20% w / w, the anthocyanin titre preferably being in a range of from 21 % to 40%, preferably from 25% to 30%, for example 25% by weight relative to the total weight of the dry extract.

[0084] For example, the mixture according to the invention may comprise a blueberry dry extract titrated at 25% in anthocyanins by HPLC, for example the blueberry dry extract sold by ACEF.

[0085] The mixture of the invention comprises, in addition to i) an extract of chamomile, ii) an extract of turmeric, iii) L- acetyl carnitine, preferably L-acetyl carnitine in the form of a salt, preferably L-acetyl carnitine hydrochloride, and iv) a blueberry extract, also v) a lipoic acid, or a salt thereof, and / or a bergamot extract.

[0086] The mixture according to the invention, Mix 1 , comprises lipoic acid or a salt thereof in oxidated or reduced form. The oxidated form (LA, cyclic disulphide) and reduced form (DHLA) constitute a potent redox pair and are capable of acting as ROS scavengers and regenerating the levels of endogenous antioxidants, such as vitamin C, vitamin E and glutathione. Both forms of the acid can neutralise different types of ROS and form complexes with different metals, depending on affinity, thus acting as chelating agents.

[0087] In the context of the present invention, lipoic acid is understood to include a lipoic acid, both in oxidated and reduced form, and / or a pharmaceutically acceptable salt thereof and / or mixtures thereof.

[0088] Preferably, the lipoic acid present in the mixture according to the invention is the microencapsulated lipoic acid present in the commercial product THIOCTIC ACID MATRIS® RETARD - AQUAGOLD sold by the firm IPS.

[0089] In the context of the present invention, the lipoic acid in the mixture (i)+(ii)+(iii)+(iv)+(v) can be replaced, in whole or in part, by a bergamot extract (Mix 2), such as, for example, a bergamot extract of the type Citrus aurantium L. var. bergamia extract.

[0090] More preferably, component (v) of the mixture of the invention is a bergamot extract (Mix 2). An example of a commercial product of Citrus aurantium L. var. bergamia extract is Bergavit™ sold by Bionap, consisting of a fruit extract obtained from Citrus aurantium Bergamia and having CAS number 89957-91-5 and maltodextrins (CAS number 9050-36-6). The bergamot extract present in said product has a flavonoid titre (neoeriocitrin, naringin, neohesperidin) of from 25% to 28% w / w as measured by HPLC.

[0091] In the context of the present invention, preferably, the lipoic acid in the mixture (i)+(ii)+(iii)+(iv)+(v) can be replaced, in whole or in part, by a bergamot extract such as, for example, a bergamot extract of the type Citrus Bergamia Risso & Poiteau extract.

[0092] Preferably, the bergamot extract in the mixture of the invention, Mix 2, comprises an amount of total flavonoids comprised from 20% to 50%, preferably comprised from 35% to 45%, more preferably from 38% to 40% by weight relative to the total weight of the extract, wherein said flavonoids comprise naringin, neohesperidin, neoeriocitrin, melitidin, and brutieridin.

[0093] Preferably, said bergamot extract in the mixture of the invention, Mix 2, comprises the active compound naringin. Preferably, naringin is present in said bergamot extract in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 28% or 29% by weight, relative to the weight of total flavonoids.

[0094] Preferably, in addition to naringin, said bergamot extract in the mixture of the invention, Mix 2, comprises the active compound neohesperidin. Preferably, neohesperidin is present in said bergamot extract in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 29% or 30% by weight, relative to the weight of total flavonoids.

[0095] More preferably, said bergamot extract in the mixture of the invention, Mix 2, comprises naringin in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 28% or 29%, by weight, relative to the weight of total flavonoids, and a neohesperidin in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 29% or 30% by weight, relative to the weight of total flavonoids.

[0096] Preferably, said bergamot extract in the mixture of the invention, Mix 2, also comprises neoeriocitrin, in addition to naringin and neohesperidin, preferably in the percentage amounts specified above.

[0097] Preferably, neoeriocitrin is present in the bergamot extract in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 26%, by weight relative to the weight of total flavonoids.

[0098] Preferably, said bergamot extract in the mixture of the invention, Mix 2, also comprises melitidin, in addition to naringin, neohesperidin and neoeriocitrin, preferably in the percentage amounts specified above. Preferably, melitidin is present in the bergamot extract in the mixture of the invention, Mix 2, in an amount by weight comprised from 0.5% to 10%, preferably comprised from 1 % to 8%, more preferably comprised from 3% to 6%, for example 5% by weight, relative to the weight of total flavonoids.

[0099] Preferably, said bergamot extract in the mixture of the invention, Mix 1 , also comprises brutieridin, in addition to naringin, neohesperidin, neoeriocitrin and melitidin, preferably in the percentage amounts specified above.

[0100] Preferably, brutieridin is present in the bergamot extract in the mixture of the invention, Mix 2, in an amount by weight comprised from 0.5% to 25%, preferably comprised from 5% to 20%, more preferably comprised from 10 to 15%, for example 11% by weight relative to the weight of total flavonoids.

[0101] More preferably, the bergamot extract in Mix 2 comprises by weight, relative to the weight of total flavonoids: from 28% to 29% naringin, e.g. 28.7%; and / or from 29% to 30%% neohesperidin, e.g. 29.2%; and / or from 24% to 26% neoeriocitrin, e.g. 25.6%%; and / or from 3% to 6% melitidin, e.g. 4.7%; and / or

[0102] - from 10 to 15% brutieridin, e.g. 11.3%.

[0103] An example of a commercial product of a Citrus Bergamia Risso & Poiteau extract is the BERGAMOT POLYPHENOLIC FRACTION™ (BPF™), for example sold by H&AD S.r.l. It is an extract from juice of the fruit of Citrus Bergamia Risso & Poiteau, comprising a titre of total flavonoids (neoeriocitrin, naringin, neohesperidin, melitidin, and brutieridin) as measured by HPLC comprised from 20 to 50%, preferably comprised from 35 to 45%, more preferably from 38% to 40%. For example, said bergamot extract has a titre of total flavonoids, as measured by HPLC, for example of 39%, of which, preferably: from 6% to 14%, preferably from 8% to 12%, more preferably 10.2% neoeriocitrin; from 6% to 14%, preferably from 8% to 13%, more preferably 11% naringin; from 6% to 14%, preferably from 8% to 13%, more preferably from 11 % to 12% neohesperidin; from 0.5% to 3%, preferably from 1 % to 2%, more preferably 2% melitidin and from 2% to 8%, preferably from 3% to 6%, more preferably from 4% to 5% brutieridin.

[0104] In other words, the total amount of flavonoids in the extract is for example 39.2% and, for example, the 39.2% is thus composed: 10.2% is the amount of neoeriocitrin, 11.4% is the amount of naringin, 11.6% is the amount of neohesperidin, 1.9% is the amount of melitidin, and 4.5% is the amount of brutieridin.

[0105] More preferably, the bergamot extract in Mix 2 comprises, by weight, relative to the weight of total flavonoids: from 6% to 40%, preferably from 15% to 35%, more preferably from 20% to 30%, for example 28% or 29%, naringin, from 6% to 40%, preferably from 15% to 35%, more preferably from 20% to 30%, for example 29% or 30% neohesperidin, from 6% to 40%, preferably from 15% to 35%, more preferably from 20% to 30%, for example 26% neoeriocitrin, from 0.5% to 10%, preferably from 1 % to 8%, more preferably from 3% to 6%, for example 5% melitidin, and from 0.5% to 25%, preferably from 5% to 20%, more preferably from 10 to 15%, for example 11% brutieridin. The bergamot extract appears as a yellow-coloured powder having particle size passing through a 60 mesh sieve and a bulk density comprised, for example, from 30 to 70 g / 100 ml, preferably comprised from 40 to 60 g / 100 ml, more preferably 55 g / 100 ml, 56 g / 100 ml, 57 g / 100 ml, 58 g / 100 ml, 59 g / 100 ml (DIN / ISO 697). Preferably, said bergamot extract has a pH comprised from 3 to 4, preferably from 3.1 to 3.8.

[0106] In the context of the present invention, the bergamot extract has a titre of total flavonoids comprised from 20% to 50%, preferably comprised from 25% to 45%, more preferably comprised from 30% to 40%, for example 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, or 39%.

[0107] For example, lipoic acid and bergamot extract can be present in the mixtures (i)+(ii)+(iii)+(iv)+(v) and in the compositions of the present invention containing said mixtures, in a ratio by weight comprised from 1 :10 to 10: 1 , preferably comprised from 1 :5 to 5:1 , more preferably comprised from 1 :3 to 3: 1 , for example 1 :1 [lipoic acid:bergamot extract].

[0108] The mixture according to the invention, Mix 1 and Mix 2, possesses an interesting activity of preventing and repairing damage to the central nervous system, preferably in counteracting motor neuron degeneration and demyelination processes.

[0109] The effect exerted by the mixture of the invention, Mix 1 and Mix 2, is not tied to the extract but to the active compounds present in the extract, preferably the active compounds present in the phytocomplex of said extracts. Nature-identical molecules of said active compounds can be present in the mixture according to the invention, Mix 1 and Mix 2 (Regulation (EC) N 1334 / 2008).

[0110] In the context of the present invention, the expression "nature-identical molecules” is understood to indicate and include molecules equal to the active compounds present in the natural extracts or phytocomplexes but produced through chemical synthesis.

[0111] For example, the active compounds naringin, neohesperidin, neoeriocitrin, melitidin, and brutieridin may be present as nature-identical molecules.

[0112] In the mixture according to the invention, Mix 2, naringin and / or neohesperidin can be present as nature-identical molecules. In other words, the mixture according to the invention, Mix 2, may comprise naringin and / or neohesperidin produced through chemical synthesis.

[0113] Preferably, the mixture according to the invention comprises or, alternatively, consists of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) a bergamot extract.

[0114] Preferably, the mixture according to the invention consists of: i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) a bergamot extract.

[0115] Preferably, the mixture of the invention, Mix 1 and Mix 2 ((i)+(ii)+(i ii)+(iv)+(v)), may also comprise at least one further ingredient (a), selected from the group comprising or, alternatively, consisting of: - a.i) at least one vitamin selected in the group comprising or, alternatively, consisting of vitamin C, vitamin E, vitamin B1 , vitamin B2, vitamin B6 and vitamin B12 or mixtures thereof,

[0116] - a.ii) L-tryptophan,

[0117] - a.iii) an extract of strawberry Fragaria,

[0118] - a.iv) an extract of Crocus sativus, or mixtures thereof a.i)+a.ii)+a.iii)+a.iv).

[0119] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also a further ingredient (a) selected from a.i), a.ii), a.iii), a.iv), and mixtures thereof.

[0120] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also a further ingredient (a.i).

[0121] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also a further ingredient (a.ii).

[0122] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also a further ingredient (a.iii).

[0123] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also a further ingredient (a.iv).

[0124] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.i) and (a.ii).

[0125] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.i) and (a.iii).

[0126] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.i) and (a.iv).

[0127] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.ii) and (a.iii).

[0128] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.ii) and (a.iv).

[0129] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.iii) and (a.iv).

[0130] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.iii) and (a.i) and (a.ii).

[0131] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.iii) and (a.i) and (a.iv).

[0132] Preferably, the mixture of the invention, Mix 1 and Mix 2, comprises, in addition to i)+ii)+iii)+iv)+ v), also (a.iii) and (a.ii) and (a.iv).

[0133] In the context of the present invention, a.i) vitamin E is understood to include any form of vitamin E, a pharmaceutically acceptable salt thereof and mixtures thereof. In the context of the present invention, a.i) vitamin C is understood to include any form of vitamin C, a pharmaceutically acceptable salt thereof and mixtures thereof.

[0134] In addition to or as a replacement for vitamin C and / or vitamin E, the mixture according to the invention, Mix 1 and Mix 2, further comprises at least one a.i) vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B6 (pyridoxine), vitamin B12 and mixtures thereof.

[0135] Preferably, the mixture of the invention comprises components i) - v) and all the vitamins C, E, B1 , B2, B6 and B12 (see example 2).

[0136] In the context of the present invention, a.ii) L-tryptophan is understood to include a tryptophan and a pharmaceutically acceptable salt thereof and mixtures thereof.

[0137] In the context of the present invention, an a.iii) extract of strawberry Fragaria is understood to include all types of strawberry extracts of Fragaria, preferably Fragaria ananassa or Fragaria L, comprising anthocyanins. Preferably, the extract of strawberry Fragaria (for example Fragaria L.) is a dry extract titrated in anthocyanins, the minimum anthocyanin titre being > 0.5% w / w, the anthocyanin titre being in a range of from 0.5% to 0.99% by weight relative to the total weight of the dry extract.

[0138] One type of strawberry extract present in the mixture according to the invention may be for example a strawberry dry extract with 0.5% anthocyanins, for example sold by ACEF, which is a Fragaria L. extract having an amount of anthocyanins (Cya-3-glu, spectrophotometry pH-Diff.) of 5.00-9.80 g / kg, and polyphenols (catechins, Folin Ciocalteu) of 20.00-46.00 g / kg.

[0139] Another type of strawberry extract present in the mixture according to the invention can be for example a strawberry dry extract called STRAWBERRY EXTRACT POWDER sold by DIANA FOOD CANADA Inc., comprising an amount of polyphenols > 2% (Folin-Ciocalteu).

[0140] In the context of the present invention, a.i) vitamin C is understood to include any form of vitamin C, a pharmaceutically acceptable salt thereof and mixtures thereof.

[0141] Preferably, the mixture according to the invention, Mix 1 and Mix 2, comprises a.iv) a saffron extract.

[0142] Crocus sativus (L), commonly called saffron, is a perennial plant belonging to the family Iridaceas. The stigma of Crocus sativus contains numerous bioactive compounds, including alpha-crocetin, crocin, picrocrocin and safranal. In the context of the present invention, an a.iv) extract of Crocus sativus is understood to include all types of Crocus sativus stigma extracts, comprising crocin and / or safranal. Preferably, the Crocus sativus extract is a dry extract titrated in crocin and / or safranal, the minimum safranal titre being >0.30% w / w and / or crocin titre being > 0.40% w / w, the safranal titre preferably being in a range of from 0.34 % to 0.45% by weight relative to the total weight of the dry extract and / or the crocin titre in a range of from 0.43% to 0.60% by weight relative to the total weight of the dry extract.

[0143] One type of saffron extract present in the mixture according to the invention can be for example the commercial product SATI EREAL® sold by Natac, which is a dry extract of saffron (Crocus sativus L.) stigmas characterised by a minimum safranal titre of 0.34% w / w as measured by spectrophotometry and a minimum crocin titre of 0.43% w / w as measured by HPLC.

[0144] Another commercial product is Affron® sold by Pharmactive Biotech Products.

[0145] Preferably, the mixture of the present invention may be in the form of a capsule, a tablet, a sachet, a stick, an orosoluble stick, an oral gel or granules.

[0146] The second aspect of the present invention relates to a composition comprising the mixture as defined above and food or pharmaceutical grade excipients and / or additives.

[0147] The composition is a pharmaceutical and / or nutraceutical composition that may be used in a method of treatment, therapeutic or non-therapeutic, with the aim of ensuring the maintenance of central nervous system function and / or in a method of preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0148] The composition of the invention is a pharmaceutical and / or nutraceutical composition for oral use, preferably a composition in a solid state, preferably formulated in dosage units. "Solid state” means that the composition can exist in the form of granules or powders. The granular or powder compositions are mixed with pharmaceutically acceptable additives and excipients to provide a final product such as, for example, a supplement product, a medical device or a pharmaceutical composition. The final product may be in pharmaceutical dosage units such as, for example, granules in a sachet, a stick, orosoluble stick, tablet, gel or capsule.

[0149] The tablets may have different shapes among those known in the field of pharmaceutical forms, such as, for example, a cylindrical or spheroidal shape. The tablets may be coated or film-coated with one or more coating or film layers capable of passing through the gastric barrier, according to known methods.

[0150] The gel capsules may consist of hard gelatine or soft gelatine or soft gel.

[0151] Preferably, the oral composition of the invention is a solid or semi-solid (gel) composition as described above. However, if desired or necessary, the composition may be formulated in liquid form, for example by dissolution or suspension in water.

[0152] The solid compositions of the invention may contain, as mentioned, physiologically acceptable conventional excipients and / or vehicles, such as diluents, bulking agents, binders, disaggregating agents, flow aids, lubricants, etc. Non-limiting examples of suitable vehicles and excipients are described in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins. Compositions of the invention may, for example, include cellulose derivatives, glucose, lactose, sucrose, gelatin, malt, rice, flour, gypsum, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, glycol, water, ethanol, and the like. The composition may also contain pH buffering reagents and wetting or emulsifying agents.

[0153] Preferably, the compositions of the present invention may be in the form of a capsule, a tablet, a sachet, a stick, an orosoluble stick, an oral gel or granules.

[0154] The composition of the invention, preferably formulated in dosage units, may be administered once or several times a day, for example once or twice a day, preferably twice a day.

[0155] Preferably, the composition comprises: from 100 to 400 mg, preferably from 150 to 350 mg, even more preferably from 175 to 200 mg, more preferably 175 mg of a dry extract of Matricaria chamomile titrated in apigenin (minimum titre > 2% w / w; the apigenin titre preferably being in a range of from 5% to 10%, more preferably 5% by weight relative to the total weight of the dry extract); from 30 to 200 mg, preferably from 50 to 160 mg, even more preferably from 60 to 100 mg of a dry extract of Curcuma longa titrated in curcuminoids (minimum titre > 30% w / w; the titre in curcuminoids preferably being in a range of from 30% to 50%, preferably from 35% to 45%, more preferably 35% by weight relative to the total weight of the dry extract); from 200 to 2,000 mg, preferably from 300 to 1 ,000 mg, more preferably from 400 to 500 mg of L-acetyl carnitine (L-carnitine and / or carnitine) or a pharmaceutically acceptable salt thereof and / or mixtures thereof; from 50 to 200 mg, preferably from 60 to 180 mg, even more preferably from 80 to 120 mg of a dry extract of Vaccinium myrtillus titrated in anthocyanins (minimum titre > 20% w / w; the anthocyanin titre preferably being in a range of from 21 % to 40%, preferably from 25% to 30% by weight relative to the total weight of the dry extract); from 100 to 1 ,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of a lipoic acid or a pharmaceutically acceptable salt thereof or mixtures thereof, or from 10 to 1 ,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of bergamot extract (wherein said bergamot extract preferably has a titre of total flavonoids (neoeriocitrin, naringin, neohesperidin, melitidin, and brutieridin) comprised from 20% to 50%, preferably from 25% to 45%, more preferably from 30% to 40% w / w), and at least one acceptable pharmaceutical or food grade additive and / or excipient.

[0156] More preferably, the composition comprises: from 100 to 400 mg, preferably from 150 to 350 mg, even more preferably from 175 to 200 mg, more preferably 175 mg of a dry extract of Matricaria chamomile titrated in apigenin (minimum titre > 2% w / w; the apigenin titre preferably being in a range of from 5% to 10%, more preferably 5% by weight relative to the total weight of the dry extract); from 30 to 200 mg, preferably from 50 to 160 mg, even more preferably from 60 to 100 mg of a dry extract of Curcuma longa titrated in curcuminoids (minimum titre > 30% w / w; the titre in curcuminoids preferably being in a range of from 30% to 50%, preferably from 35% to 45%, more preferably 35% by weight relative to the total weight of the dry extract); from 200 to 2.000 mg, preferably from 300 to 1 ,000 mg, more preferably from 400 to 500 mg of L-acetyl carnitine (L-carnitine and / or carnitine) or a pharmaceutically acceptable salt thereof and / or mixtures thereof; from 50 to 200 mg, preferably from 60 to 180 mg, even more preferably from 80 to 120 mg of a dry extract of Vaccinium myrtillus titrated in anthocyanins (minimum titre > 20% w / w; the anthocyanin titre preferably being in a range of from 21 % to 40%, preferably from 25% to 30% by weight relative to the total weight of the dry extract); from 10 to 1,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of bergamot extract (wherein said bergamot extract preferably has an amount of total flavonoids (neoeriocitrin, naringin, neohesperidin, melitidin, and brutieridin) comprised from 20% to 50%, preferably from 25% to 45%, more preferably from 30% to 40% w / w) and at least one acceptable pharmaceutical or food grade additive and / or excipient.

[0157] Preferably, in addition to components I) - v), the composition comprises: from 50 to 500 mg, preferably from 80 to 400 mg, even more preferably from 125 to 200 mg of vitamin C; and / or from 2 to 50 mg, preferably from 5 to 30 mg, even more preferably from 9 to 20 mg of vitamin E; and / or from 1 to 50 mg, preferably from 3 to 30 mg, even more preferably from 6 to 20 mg, for example 6.25 mg of vitamin B1; and / or from 1 to 50 mg, preferably from 3 to 30 mg, even more preferably from 6 to 20 mg of vitamin B2; and / or from 0.5 to 30 mg, preferably from 1 to 15 mg, even more preferably from 2 to 6 mg, for example 2.38 mg of vitamin B6; and / or from 1 to 50 g, preferably from 3 to 30 pg, even more preferably from 6 to 20 pg, for example 6.25 pg of vitamin B12; and / or from 5 to 300 mg, preferably from 10 mg to 200 mg, more preferably 15 mg of a Crocus sativus dry extract titrated in crocin and / or safranal (the minimum safranal titre being >0.30% w / w and / or crocin titre being >0.40% w / w, the safranal titre preferably being in a range of from 0.34 % to 0.45% by weight relative to the total weight of the dry extract and / or the crocin titre in a range of from 0.43% to 0.60% by weight relative to the total weight of the dry extract) and / or from 1 to 500 mg, preferably from 10 to 300 mg, even more preferably from 50 to 200 mg of L-tryptophan, for example 150 mg and / or from 2 to 1 ,000 mg, preferably from 10 to 500 mg, even more preferably from 80 to 180 mg of a strawberry dry extract, for example an extract of strawberry Fragaria (for example Fragaria L) titrated in anthocyanins (minimum titre > 0.5% w / w, the anthocyanin titre preferably being in a range of from 0.5% to 0.99% by weight relative to the total weight of the dry extract), and at least one acceptable pharmaceutical or food grade additive and / or excipient.

[0158] The dosages indicated above are to be understood as a dosage unit and not as the daily amount.

[0159] The compositions of the invention can be prepared by mixing the single components, and any conventional excipients and / or vehicles.

[0160] In particular, both the mixture of the invention and the compositions comprising them are easy to prepare and inexpensive, considering that they can be prepared using the apparatus and preparation processes known to the person skilled in the art and in pharmaceutical practice or in the sector of nutritional supplements.

[0161] The daily dosage depends on the patient's health status, weight, gender and age. In general, the compositions of the invention are well tolerated and can be administered once or several times a day, preferably twice a day, even over an extended period of time. A third aspect of the present invention relates to the mixture, Mix 1 and Mix 2, according to the invention and / or the composition comprising said mixture for use as a medicament.

[0162] The Applicant has found that the mixture of the invention and / or the pharmaceutical and / or nutraceutical compositions containing them are useful for protecting central nervous system function and counteracting motor neuron degeneration and demyelination processes and / or in a method of preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0163] According to another of the aspects thereof, the invention relates to the use of the mixture according to the invention and / or the composition comprising it in a method of treatment, therapeutic or non-therapeutic, with the aim of maintaining central nervous system function and / or counteracting motor neuron degeneration and demyelination processes and / or in a method of preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS). More preferably, the mixture according to the invention and / or the composition comprising said mixture are used for the treatment of Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0164] The present description relates to a method for preventing and repairing damage to the central nervous system, preferably for counteracting motor neuron degeneration and demyelination processes and / or preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS), the method comprising administering the mixture and / or the composition of the invention to an individual in need thereof, preferably in a dose effective for the purpose.

[0165] Advantageously, the mixture and / or the composition according to the invention can be administered on their own or together with drugs and / or food supplements commonly used for preventing and repairing damage to the central nervous system, preferably to counteract motor neuron degeneration and demyelination processes.

[0166] For example, the mixture and / or the composition according to the invention can be administered together with drugs commonly used to treat ALS and / or MS.

[0167] Preferably, the mixture and / or the composition of the invention are for use in a method of preventing and / or treating Amyotrophic Lateral Sclerosis (ALS).

[0168] Preferably, said mixture and / or said composition can be administered together with at least one drug conventionally used for the treatment of Amyotrophic Lateral Sclerosis (ALS), preferably together with Riluzole. In fact, the data reported in the experimental section demonstrate that it is possible to combine the mixture according to the invention with treatment with the drug of first choice, Riluzole, because in all cases the biological effectiveness of the drug is enhanced by the combination with the mixture according to the invention (see Example 6 - Phase III). Preferably, by means of the composition of the present invention, a daily dose of a dry extract of Matricaria chamomile titrated in apigenin (titre 5% w / w) equal to 350 mg (17.5 mg of apigenin) is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0169] Preferably, by means of the composition of the present invention, a daily dose of a dry extract of Curcuma longa titrated in curcuminoids (titre 35% w / w) equal to 120 mg (42 mg of curcuminoids) is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick. Preferably, by means of the composition of the present invention, a daily dose of L-acetyl carnitine or L-carnitine or carnitine equal to 800 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0170] Preferably, by means of the composition of the present invention, a daily dose of a dry extract of Vaccinium myrtillus titrated in anthocyanins (titre 25% w / w) equal to 160 mg (40 mg of anthocyanins) is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick. Preferably, by means of the composition of the present invention, a daily dose of a strawberry dry extract, for example an extract of strawberry Fragaria titrated in anthocyanins (titre = 0.5% w / w) equal to 160 mg is administered for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0171] Preferably, by means of the composition of the present invention, a daily dose of a lipoic acid or a pharmaceutically acceptable salt thereof or mixtures thereof equal to 600 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0172] Preferably, by means of the composition of the present invention, a daily dose of bergamot extract equal to 600 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0173] Preferably, by means of the composition of the present invention, a daily dose of vitamin C equal to 250 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0174] Preferably, by means of the composition of the present invention, a daily dose of vitamin E equal to 18 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0175] Preferably, by means of the composition of the present invention, a daily dose of vitamin B1 equal to 12.5 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0176] Preferably, by means of the composition of the present invention, a daily dose of vitamin B2, for example equal to 12.5 mg, is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0177] Preferably, by means of the composition of the present invention, a daily dose of vitamin B6, for example equal to 4.75 mg, is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick. Preferably, by means of the composition of the present invention, a daily dose of vitamin B12, for example equal to 12.55 pig, is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0178] Preferably, by means of the composition of the present invention, a daily dose of a dry extract of Crocus sativus titrated in crocin and / or safranal as defined above equal to 30 mg is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0179] Preferably, by means of the composition of the present invention, a daily dose of L-tryptophan, for example equal to 300 mg, is administered, for example, by means of one administration of a sachet, or two administrations of a tablet or sachet or orosoluble stick or gel stick.

[0180] Furthermore, the mixture of the invention and the pharmaceutical and / or nutraceutical compositions containing it, due to the presence of natural ingredients, do not have significant adverse effects, are free of side effects and have high tolerability, and can be administered to a broad category of individuals.

[0181] Finally, the mixture and / or the compositions of the invention are easy to prepare and inexpensive.

[0182] Some representative embodiments of the compositions of the invention are given in the Experimental Section below solely by way of illustration and are in no way limiting.

[0183] As will be shown in the Experimental Section that follows and by the figures illustrating the results of the experimental assays conducted, the mixture of the invention ("MIX 1” in the examples and figures) has been demonstrated to be effective in protecting central nervous system function, preferably in counteracting motor neuron degeneration and demyelination processes and / or in a method of preventing and / or treating neurodegenerative diseases affecting motor neurons and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

[0184] EXAMPLES

[0185] Example 1

[0186] Composition in the form of a tablet or sachet or orosoluble stick or gel stick (1 dosage form twice a day) containing:

[0187] Table 1 together with food or pharmaceutical grade excipients and / or additives. The lipoic acid can be replaced by bergamot extract (1 :1 replacement).

[0188] Example 2 Composition in the form of a tablet or sachet or orosoluble stick or gel stick (1 dosage form twice a day) containing:

[0189] Table 2 together with food or pharmaceutical grade excipients and / or additives.

[0190] Example 3 Composition in the form of a tablet or sachet or orosoluble stick or gel stick (1 dosage form twice a day) containing:

[0191] Table 3 together with food or pharmaceutical grade excipients and / or additives.

[0192] Example 4 Composition in the form of a tablet or sachet or orosoluble stick or gel stick (1 dosage form twice a day) containing:

[0193] Table 4 together with food or pharmaceutical grade excipients and / or additives.

[0194] The activity of the mixture and of the compositions containing them was assessed on the basis of various experimental in vitro assays. Example 5 - In vitro study

[0195] 5.1. Experimental protocol

[0196] An in vitro study was conducted with the aim of investigating the biological action of a mixture according to the invention in counteracting the mechanisms that lead to motor neuron degeneration and trigger axon demyelination. The experiments were conducted using a model of HCN-2 motor neurons, i.e. a cell line of cortical neurons under physiological conditions. The active compounds of interest were tested in vitro, their concentrations were identified based on data of the literature and following a viability study, in which it was verified that the preselected range of concentration was not cytotoxic.

[0197] The final definitive concentration chosen for each single active compound, used in the subsequent experiments and to compose the mixture, is the one that demonstrated the greatest effectiveness in increasing cell viability versus the control (represented by untreated cells in basal conditions). The following mixture was then prepared and tested:

[0198] MIX 1 (according to the invention): L-acetyl carnitine hydrochloride 1 pM; extract of chamomile containing 5% apigenin 10 pg / mL; extract of Vaccinium myrtillus containing 250 pg / mL anthocyanins; lipoic acid 10 pM; extract of turmeric (CurQfen®) 100 pM.

[0199] The study was then developed in four different phases.

[0200] PHASE I

[0201] The mixture of interest must exert its effectiveness at the level of the central nervous system. Therefore, the first phase of experiments was aimed at verifying that the mixture can safely reach the target of action. A three- dimensional co-culture model of the blood-brain barrier was thus set up, composed of astrocytes and HUVECs (vein endothelial cells); on the one hand, the passage velocity and efficiency of the single active compounds and of the mix were calculated and, on the other hand, the membrane integrity was checked by studying the expression of the markers MarvelD3 and claudin 5. In this manner, it was possible to demonstrate that the mixture of the invention has a significantly greater action (p<0.05) compared to the single active compounds and to the drug used as a reference, Riluzole, both in terms of absorption rate and in terms of safety.

[0202] PHASE II

[0203] The second phase of the protocol regarded the assessment of any possible toxicity of the single compounds, the Riluzole and the mixtures, at the preselected concentrations, on the motor neuron model, HCN-2 cell line, in order to rule out a cytotoxic effect under physiological conditions. Based on the analyses performed, it was demonstrated that the tested substances are capable of stimulating cell viability, thus ruling out any cytotoxic effect and confirming the safety of the tested mixture. Specifically, the tested mixture increases cell viability without altering neuronal homeostasis (p<0.05) (Figure 1). PHASE III

[0204] Then, in Phase III, the protocol provided for a study of the biological activity of the single components and of the mixture of the invention. The mixture according to the present invention has the objective of counteracting the molecular mechanisms which trigger motor neuron degeneration and the axon demyelination process. Therefore, the subsequent experiments provided for the use of a toxic stimulation with glutamate at 5 mM, so as to mimic the pathological condition of neurodegeneration in vitro and the biological action of the mixture was investigated by testing different markers involved in neurodegeneration.

[0205] PHASE IV

[0206] In the last phase of the experiments, any possible interference between the single active compounds and mixtures and the drug generally used in the case of motor neuron degeneration and axon demyelination, namely Riluzole, was investigated.

[0207] 5.2. Results

[0208] Ability to reduce glutamate-induced mortality

[0209] First, it was verified whether the single compounds and the mixture had the ability to reverse the process of cell mortality that arises as a result of pretreatment with the toxic stimulation produced by glutamate. As shown in Figure 2, all the single substances have an ability to significantly reduce glutamate-induced mortality, but only the mixture is capable of bringing the values back to a basal level. Specifically, table 5 shows the % value of the influence that each raw material has on viability. Taking a numerical sum of the % values, one would even obtain a negative value, i.e. a reduction in viability. In contrast, Mix 1 , with its synergistic action, has the effect of increasing viability above the values of the healthy control (table 5 and Figure 2).

[0210] This method for demonstrating the synergy of action of multiple compounds is widely used in the scientific literature and directly reflects the definition of pharmacological synergism, according to which "the final effect is greater than the algebraic sum of the individual responses of two substances that exert the same biological effect with two identical or different mechanisms”. An example of scientific literature where this method is used to demonstrate synergy is the paper published in Antioxidants in 2022: Matacchione, G.; Valli, D.; Silvestrini, A.; Giuliani, A.; Sabbatinelli, J.; Giordani, C.; Coppari, S.; Rippo, M.R.; Albertini, M.C.; Olivieri, F. "Curcumin, Polydatin and Quercetin Synergistic Activity Protects from High-Glucose-lnduced Inflammation and Oxidative Stress”. Antioxidants 2022, 11 , 1037. Table 5: % of neuroprotective activity performed by the single active compounds, the numerical sum thereof and Mix 1

[0211] Inhibition of ROS Production

[0212] Neurodegeneration is also caused by an excessive production of oxygen radicals induced by pretreatment with glutamate. As shown in Figure 3, ROS production is inhibited by the substances under examination (p<0.05), which, if combined, are capable of having a protective effect by acting on the antioxidant capacities of neuronal cells attempting to restore homeostasis (p<0.05). Furthermore, Mix 1 demonstrates a synergistic action compared to the single agents, since, as demonstrated in table 6, treatment with each single component still results in the production of a certain % amount of oxidative stress compared to the healthy control, and the sum of the activities of the single components would lead to a positive % of ROS production. In contrast, only the mixture, with its synergistic action, is capable of completely reversing this process (table 6 and Figure 3).

[0213] Table 6: % of oxidant activity performed by the single active compounds, the numerical sum thereof and Mix 1.

[0214] Ability to inhibit nitric oxide production

[0215] The analysis on NO production confirms its central role in neurodegenerative processes, as it causes the formation of highly reactive species which increase the levels of oxidative stress, thereby activating the apoptotic mechanisms of CNS cells. Therefore, the inhibition of nitric oxide production by the tested substances confirms their potential usefulness (p<0.05); in this case as well, MIX 1 synergistically increases the beneficial effect of the single agents (Table 7 and Figure 4). Table 7: % of nitrosative activity performed by the single active compounds, the numerical sum thereof and Mix 1

[0216] Oxidative and nitrosative stress also contribute to lipid and protein peroxidation. By testing MIX 1 on these two factors, it was in fact demonstrated that the mixture according to the invention is more effective than the single active compounds in protecting against this cell damage (p<0.05).

[0217] Levels of glutathione (GSH) production

[0218] In the physiopathology of some diseases involving neurodegeneration and demyelination, as in the case of Amyotrophic Lateral Sclerosis and Multiple Sclerosis, the imbalance in glutathione synthesis and glutamate metabolism plays a pivotal role. In these patients, in fact, a condition of excess glutamate comes to be created, which causes excitotoxicity, along with excess glutathione, which alters the balance of the antioxidant defences of nerve cells. Therefore, when these two disease markers were analysed, it emerged that the antioxidant mechanism is restored, notwithstanding the damage induced by glutamate, both by the single active compounds, and by the mixture, with a greater effectiveness of the mixture, which shows to be significant compared to the single compounds. The same result is confirmed also as far as the release of glutamate is concerned (Figure 5). Furthermore, as may be noted from table 8, the mixture's action on GSH is of a synergistic type, as compared with effectiveness of the active compounds taken individually.

[0219] Table 8: % of glutathione production induced by the single active compounds, the numerical sum thereof and Mix 1

[0220] Increase in intracellular calcium

[0221] Closely correlated to glutamate excitotoxicity, there is an increase in intracellular calcium, which triggers a chain of events that fuel neurodegeneration. In this case as well, the mixture showed to be more effective, when compared to the single active compounds making it up, in reducing the calcium levels resulting from glutamate-induced damage (p<0.05).

[0222] Expression of some markers of neurodegeneration caused by glutamate-induced damage

[0223] In order to assess the effectiveness of the single compounds and of the mixture in a condition of neurodegeneration, the principal markers that are altered in this condition, namely ADAR2, SOD1 , TDP43 and TUBB4A, were investigated. In fact, the accumulation of TDP-43 in axons and in the neuromuscular junctions negatively affects the synthesis of the proteins involved in the synaptic function, whilst low levels of ADAR2 trigger a neurodegenerative process. In contrast, SOD1 plays a key role in antioxidant mechanisms and TUBB4A codes for the synthesis of beta tubulin, a protein of which there is a deficiency in the case of axon hypomyelination.

[0224] In the analysis performed on all the aforementioned markers, Mix 1 showed to be statistically more active compared to the single compounds (p<0.05).

[0225] Specifically, Figures 6 A and B and table 9 show the synergism of the mixture compared to the activity of the single compounds.

[0226] Table 9: % of activity on SOD and ADAR2 production of the single active compounds, the numerical sum thereof and MIX 1

[0227] PHASE IV

[0228] In conclusion, in the last phase of the experiments, any possible interference between the single active compounds and mixtures and the drug generally used in the case of motor neuron degeneration and axon demyelination, namely Riluzole, was investigated.

[0229] All the above-described experiments were thus replicated, adding Riluzole at 3 different times of the experimental protocol. In all cases the experimental model was induced with damage caused by pretreatment with 5 mM of glutathione, after which three scenarios were set up:

[0230] Damage with glutathione followed by treatment with Riluzole and after 1h of treatment with the active compounds and the mixture (this simulates the condition of a patient with ALS undergoing treatment with Riluzole, who starts also taking the supplement of the formulation).

[0231] Damage with glutathione, followed by co-treatment with Riluzole and the single active compounds and the mixture (this simulates the condition of a patient who, upon being diagnosed, undergoes both the pharmacological treatment and treatment with the supplement of the formulation).

[0232] Damage with glutathione, followed by treatment with the active compounds and the mixture, and subsequently, after 1 h of treatment, Riluzole is added (this simulates the condition of a patient who, pending a certain diagnosis, starts taking the supplement and then also has recourse to treatment with the drug).

[0233] As may be deduced from Figures 7 A-C and Table 10, all the substances and the mixture under examination are able to improve cellular conditions if administered simultaneously with or before or after Riluzole. This effect is synergistically amplified when the substances are combined in the mixture, thereby stimulating mitochondrial wellbeing (p<0.05). These data suggest that the active compounds combined in the mixture thereof are capable of activating survival mechanisms in a more effective and synergistic manner than when used individually (p<0.05 vs the single substances) and compared to the drug of reference (p<0.05).

[0234] Figures 8 A-C show the results derived from the analysis of ROS production under the three experimental conditions described above. In all cases it was confirmed that the substances under examination are capable of counteracting oxidative stress and, if combined in the mixture, they are able to have a greater protective effect; in particular, Mix 1 shows to be synergistic compared to the active compounds making it up (p<0.05) (table 10). The antioxidant defence mechanism that is established does not interfere with the administration of Riluzole; furthermore, in this case as well, the mixture shows to be more effective than Riluzole in reducing oxidative stress.

[0235] As regards nitric oxide production, the same results as obtained for ROS are confirmed (Figures 9 A-C). Mix 1 shows to be more effective than the drug of reference and synergistic compared to the single active compounds (table 10).

[0236] Table 10: % of activity of the single components, the mathematical sum thereof and Mix 1 on cell viability and ROS and NO production under the three conditions of pre-administration, co-administration and postadministration of Riluzole

[0237] As regards the presence of lipid and protein peroxidation under the three experimental conditions, the previously obtained data are reconfirmed. The substances examined can decrease the conditions of peroxidation that are triggered as a result of glutamate-induced damage. The mixture shows to be more effective than both the single agents and the drug Riluzole, and shows synergistic activity compared to the single active compounds under all the conditions considered (Figure 10 A-F and table 11).

[0238] Table 11 : % of activity of the single components, the mathematical sum thereof and Mix 1 on the parameters specified in Figure 10 (lipid and protein peroxidation) under the three conditions of pre-administration, co- administration and post-administration of Riluzole t By analyzing he release of glutamate that causes neurodegenerative excitotoxicity, it has been demonstrated that treatment with the single agents and the mixture is capable of effectively countering its accumulation. In particular, the mixture is capable of restoring the physiological levels of glutamate independently of the pharmacological treatment and more effectively than the latter.

[0239] This fact was also confirmed by analysing the release of calcium at an intracellular level. As may be deduced from Figures 11 A-C, Mix 1 is able to reduce calcium accumulation both independently of the drug's presence, and more effectively than the latter, and synergistically compared to the single active compounds making it up (Table 12).

[0240] Table 12: % of activity of the single components, the mathematical sum thereof and Mix 1 on the parameters specified in Figure 11, i.e. the concentration of intracellular calcium under the three conditions of preadministration, co-administration and post-administration of Riluzole

[0241] Lastly, in Figures 12 A-F the biomarkers already mentioned previously, and which are altered in the case of motor neuron degeneration and axon demyelination, were taken into consideration. In this case as well, the effectiveness of the single components and of the mixture thereof is not influenced by the pharmacological treatment and, as further confirmation, Mix 1 showed to be more effective than Riluzole in all cases and under most conditions showed a synergistic action compared to the single components (Table 13).

[0242] Table 13: % of activity of the single components, the mathematical sum thereof and Mix 1 on the parameters specified in Figure 12, i.e. the markers SOD and ADAR2 typical of motor neuron degeneration and demyelination under the three conditions of pre-administration, co-administration and post-administration of Riluzole

[0243] Conclusions

[0244] In conclusion, based on the data obtained, it is possible to highlight that the mixture of the present invention (MIX 1):

[0245] 1) is safe for the neuronal cell population of reference

[0246] 2) is capable of passing through the blood-brain barrier in an efficient and safe manner, in order to be able to perform its biological action at the level of the central nervous system

[0247] 3) has neuroprotective action, increasing cell viability 4) has antioxidant action, both in terms of reducing ROS and NO production and lipid and protein peroxidation, and in terms of increasing antioxidant defences by means of its direct action on glutathione.

[0248] 5) is capable of counteracting the cause of the neurodegeneration, by counteracting the increase in glutamate and intracellular calcium 6) is capable of acting directly on markers that are altered in the event of disease, such as SOD1,

[0249] ADAR2, TDP43 and TUBB4A, in particular on the markers SOD1 and ADAR2.

[0250] 7) has no interferences with the drug Riluzole, but on the contrary shows a significantly greater action than the latter.

[0251] 8) was demonstrated to be synergistic compared to the active compounds making it up in the majority of the tests carried out.

[0252] Example 6 - Effectiveness of a composition comprising v) lipoic acid (Mix 1) vs a composition comprising v) bergamot (Mix 2)

[0253] The Applicant compared the effectiveness of a composition comprising v) lipoic acid (Mix 1) and a composition comprising v) bergamot (Mix 2) in counteracting the molecular mechanisms that trigger motor neuron degeneration and the axon demyelination process.

[0254] 6.1. Materials and methods

[0255] The study was divided into 3 phases:

[0256] PHASE I

[0257] The mixture of interest must exert its effectiveness at the level of the central nervous system. Therefore, the first phase of the experiments was aimed at verifying that the mixture can safely reach the target of action. Therefore, a three-dimensional co-culture model of the blood-brain barrier was set up, composed of astrocytes and HUVECs (vein endothelial cells).

[0258] PHASE II

[0259] The objective of this phase was to verify whether the single active compounds and the mixture are capable of counteracting the molecular mechanisms underlying the disease through:

[0260] - An analysis of effectiveness under conditions of damage induced by pretreatment with glutamate 5 mM

[0261] - An analysis of mitochondrial metabolism

[0262] - An analysis of oxidative stress through analyses of ROS, RNS, protein peroxidation and MDA

[0263] - An analysis of antioxidant mechanisms by evaluating GSH / GSSG

[0264] - An analysis of glutamate release

[0265] - An analysis of calcium release

[0266] - An analysis of specific markers (SOD1 , TPD43 and ADAR2)

[0267] PHASE III

[0268] The aim of this phase was to verify a possible enhancement or interference of the mixture with the drug of reference Riluzole. In particular, in this phase an evaluation was made of the effectiveness of the formulations in decreasing the adverse effects of Riluzole and enhancing its benefits: the analyses performed in this phase were replicated in Phase III under different experimental conditions: pre-Riluzole; post-Riluzole; and co-stimulation.

[0269] In Phases II and III cellular damage was induced by means of pretreatment with glutamate for every experimental condition. For the present experimental part, the single active compounds were tested under the same conditions as in the experiments conducted previously:

[0270] L-Acetyl carnitine hydrochloride 1 pM; Extract of chamomile titrated in 5% Apigenin 10 pg / mL; Blueberry extract titrated in anthocyanins 250 pg / mL; extract of turmeric (CurQfen®) 100pM; Lipoic acid 10pM or Bergamot d.e. (BPFTM) 10 pM.

[0271] Mix 1 : Lipoic acid - L-acetyl-carnitine - Chamomile - Blueberry - Turmeric

[0272] Mix 2: Bergamot - L-acetyl-carnitine - Chamomile - Blueberry - Turmeric

[0273] 6.2. Results

[0274] PHASE I

[0275] The tests on the blood-brain barrier (BBB) showed that the substances are capable of going beyond the bloodbrain barrier without altering the barrier exchanges, with a maximum peak at 12h (p<0.05), in the case of both the single substances and the combinations. The combined effect of the substances shows significant differences compared to Riluzole and a significantly absorption of MIX 2 compared to the active compounds making it up, but between the two combinations (MIX 1 and MIX 2) there were no statistically significant differences in terms of absorption except at 6h (p<0.05) when Mix 2 showed to be significantly more absorbed than Mix 1 (Figure 13).

[0276] The tests on the BBB tight junctions highlighted a total safety of the passage of the single active compounds and the mixture through the barrier. The activity of the two markers Marveled and claudin 5 in fact increases, demonstrating that the structural integrity was maintained upon the passage of the substances, both on their own and in combination (p<0.05 vs the control).

[0277] In particular, claudin 5 is involved in the selectivity of the membrane pores and is tied to its stability upon the passage of molecules. Marveled ensures the maintenance of the barrier upon the passage of macromolecules, allowing the release of effectors downstream. It was observed that MIX 2 shows to be statistically superior to the active compounds making it up and shows activity comparable to that of MIX 1 (Figures 14 A and B).

[0278] PHASE II

[0279] The analysis of cell viability demonstrates that the damage induced by glutamate at 5 mM is capable of reducing cell viability compared to the untreated control (p<0.05). All the substances under examination are capable of reducing the glutamate-induced damage (p<0.05) and this effect is amplified by the two mixtures (p<0.05) (Figure 15 and Table 14). Mix 2 shows synergism compared to the single substances making it up and an equivalence in terms of activity is demonstrated between the two mixtures under examination. Table 14: Analysis of cell viability

[0280] The damage induced by glutamate at 5 mM increases the levels of oxidative stress, which are effectively counteracted by the substances under examination (p<0.05), which in combination are capable of exerting a greater and synergistic effect by acting on neuronal cell antioxidant capacities, thereby favouring the restoration of homeostasis (Table 15 and Figure 16).

[0281] Table 15: oxidative stress

[0282] The analysis on NO Production confirms the potential usefulness of the substances under examination (p<0.05). The mixtures increase the beneficial effect of the single agents in a synergistic manner and in this case MIX 2 has a significant effect vs MIX 1 (p<0.05) (Table 16 and Figure 17).

[0283] Table 16: NO Production

[0284] The analysis of lipid and protein peroxidation processes confirms the previously obtained data. The substances under examination can decrease the lipid and protein peroxidation that is triggered as a result of damage with glutamate at 5 mM (p<0.05) (Table 17 and 18). In particular, Mix 2 demonstrates synergism compared to the active compounds making it up in counteracting lipid and protein peroxidation, with a level of activity comparable to that of Mix 1.

[0285] Table 17: Lipid peroxidation Table 18: Protein peroxidation

[0286] In the ALS model, the glutamate levels show to be altered. The data obtained reveal that the active single compounds and mixtures are capable of repairing the glutamate-induced damage and that the release of glutamate decreased following treatment in a statistically significant manner (p<0.05 vs glutamate; MIX p<0.05 vs the respective single components) (Tables 19 and 20 and Figure 18 and 19). Furthermore, Mix 2 shows synergism compared to the active compounds making it up and is equivalent in activity to the other mixture studied.

[0287] Table 19: GSH quantification

[0288] Table 20: Glutamate release

[0289] The dysfunction and loss of motor neurons in ALS have been attributed to various mechanisms, including an increase in intracellular calcium, which causes an alteration of motor neuron antioxidant systems; the alteration of SOD activity, for example, is one of the most well-known markers.

[0290] Figures 20 and 21 show that the single agents and the mixtures are capable of reducing calcium and SOD levels versus the glutamate-induced damage (p<0.05); in particular, in both cases MIX 2 shows synergism compared to the single active compounds and has an activity comparable to that of MIX 1 insofar as calcium is concerned, and statistically greater than that of MIX 1 as regards SOD activity.

[0291] Table 21 : Calcium release Table 22: SOD activity

[0292] Furthermore, the Applicant took into consideration two ALS markers: TDP43 and ADAR2.

[0293] The accumulation of TDP43 in axons and in neuromuscular junctions negatively impacts on the synthesis of the proteins involved in the synaptic function. As regards ADAR2, the inhibition thereof is indicative of an ongoing neurodegenerative process. In both cases, treatment with glutamate alters the activity of both markers, and this alteration is only partly counteracted by treatment with the single active compounds, whereas the mixtures show to be more effective, with Mix 2 showing to be synergistic compared to the active compounds making it up and statistically better (p<0.05) than Mix 1 insofar as TDP43 is concerned (Figures 22 and 23 and Tables 23 and 24).

[0294] Table 23: TDP43 activity

[0295] Table 24: ADAR2 activity

[0296] PHASE III

[0297] All the abovementioned experiments were then replicated, adding Riluzole at 3 different times of the experimental protocol. In all cases the experimental model was induced with the damage caused by pretreatment with 5 mM of glutathione, after which three scenarios were set up:

[0298] Damage with glutathione followed by treatment with Riluzole and after 1h of treatment with the active compounds and the mixtures (this simulates the condition of a patient with ALS undergoing treatment with Riluzole, who starts also taking the supplement of the formulation);

[0299] Damage with glutathione, followed by co-treatment with Riluzole and the single active compounds and mixtures (this simulates the condition of a patient who, upon being diagnosed, undergoes both the pharmacological treatment and treatment with the supplement of the formulation);

[0300] Damage with glutathione, followed by treatment with the active compounds and mixtures, and subsequently, after 1 h of treatment, Riluzole is added (this simulates the condition of a patient who, pending a certain diagnosis, starts taking the supplement and then also has recourse to treatment with the drug). The analysis of cell viability demonstrates that the damage induced by glutamate at 5 mM is capable of reducing cell viability. All the substances under examination are capable of improving cellular conditions irrespective of the time at which they are administered (pre-Riluzole, co-Riluzole and post-Riluzole). This effect is amplified in the effectiveness of the mixtures, where Mix 2 in particular shows synergism compared to the active compounds making it up and equivalent to that of Mix 1 (Figure 24 A-C). Furthermore, if administered after Riluzole, Mix 2 is statistically (p<0.05) more effective in improving cell viability compared to Mix 1 . The analysis of ROS production demonstrates that the damage induced by glutamate at 5 mM is capable of inducing oxidative stress. All the substances under examination are capable of improving cellular conditions irrespective of the time at which they are administered (before, after or in co- administration with Riluzole). This effect is amplified in the effectiveness of the mixtures, where Mix 2 in particular shows synergism compared to the active compounds making it up and equivalent to that of Mix 1 (Table 25).

[0301] Table 25: oxidative stress analysis of the components and mixtures in combination with Riluzole

[0302] 25.1 Pre-Riluzole

[0303] 25.2 Co-Riluzole

[0304] 25.3 Post-Riluzole

[0305] The analysis of NO Production demonstrates that the damage induced by glutamate at 5 mM is capable of reducing the production thereof. All the substances under examination are capable of improving cellular conditions irrespective of the time at which they are administered (before, after or in co-administration with Riluzole). This effect is amplified in the effectiveness of the mixtures, where Mix 2 in particular shows synergism compared to the active compounds making it up and equivalent to that of Mix 1 (Table 26).

[0306] Table 26: nitric oxide analysis of the components and mixtures in combination with Riluzole 26.1 Pre-Riluzole

[0307] 26.2 Co-Riluzole

[0308] 26.3 Post-Riluzole

[0309] The analysis of lipid peroxidation confirms the previously obtained data. All the substances under examination are capable of decreasing the lipid peroxidation triggered by the glutamate-induced damage irrespective of the time at which they are administered (before, after or in co-administration with Riluzole). This effect is amplified in the effectiveness of the mixtures, where Mix 2 in particular shows synergism compared to the active compounds making it up and equivalent to that of Mix 1. Furthermore, Mix 2 shows to be statistically more effective than Mix 1 if it is administered after Riluzole (p<0.05) (Figure 25 A-C; Table 27).

[0310] Table 27: lipid peroxidation analysis of the components and of the mixtures in combination with Riluzole

[0311] 27.1 Pre-Riluzole 27.2 Co-Riluzole

[0312] 27.3 Post-Riluzole The analysis of lipid peroxidation confirms the previously obtained data. All the substances under examination are capable of decreasing the lipid peroxidation triggered by the glutamate-induced damage irrespective of the time at which they are administered (before, after or in co-administration with Riluzole). This effect is amplified in the effectiveness of the mixtures, where Mix 2 in particular shows synergism compared to the active compounds making it up and equivalent to that of Mix 1. Furthermore, Mix 2 shows to be statistically more effective than Mix 1 if it is administered after Riluzole (p<0.05) (Figure 26 A-C; Table 28).

[0313] Table 28: protein peroxidation analysis of the components and mixtures in combination with Riluzole

[0314] 28.1 Pre-Riluzole

[0315] 28.2 Pre-Riluzole

[0316] 28.3 Post-Riluzole

[0317] The results relating to the glutamate levels show that the release of glutamate decreased following treatment with the single substances and the respective mixtures (p<0.05). The time at which the active compounds or mixtures are administered (before, after or in co-administration with Riluzole) is uninfluential.

[0318] When administered after Riluzole, Mix 2 decreases the release of glutamate to a statistically greater extent than Mix 1 (p<0.05) (Figure 27 A-C; Table 29).

[0319] Table 29: glutamate release with the components and mixtures in combination with Riluzole 29.1 Pre-Riluzole

[0320] 29.2 Co-Riluzole

[0321] 29.3 Post-Riluzole

[0322] As regards the calcium levels, the single agents and the mixtures are capable of reducing the calcium levels versus the glutamate-induced damage (p<0.05); in particular, Mix 2 shows synergism compared to the single active compounds making it up, and shows to be statistically superior also compared to MIX 1 under all conditions (p<0.05) (Table 30). Table 30: calcium levels of the components and mixtures in combination with Riluzole

[0323] 30.1 Pre-Riluzole

[0324] Figures 28 and 29 show the levels of TDP43 and ADAR2 after glutamate-induced damage and treatment with the single substances and the mixtures Mix 1 and Mix 2 before, after and during treatment with Riluzole.

[0325] The data reported in Figure 28 show that the single active compounds and the mixtures are capable of reducing TDP43 activity versus the glutamate-induced damage (p<0.05); in particular, MIX 2 succeeds in reducing the damage in a more effective and synergistic manner compared to the respective single active compounds (p<0.05). All agents show to be effective irrespective of the time at which Riluzole is administered. Furthermore, the effect of MIX 2 shows to be statistically greater than that of MIX 1 under all conditions (p<0.05).

[0326] The data reported in Figure 29 A-C show that the single active compounds and the mixtures are capable of increasing ADAR2 activity versus the glutamate-induced damage (p<0.05); in particular, MIX 2 succeeds in reducing the damage in a more effective and synergistic manner compared to the respective single active compounds (p<0.05). All agents show to be effective irrespective of the time at which Riluzole is administered. Furthermore, the effect of MIX 2 shows to be statistically greater than that of MIX 1 if it is administered simultaneously with Riluzole (p<0.05).

[0327] Also as regards SOD activity, the single active compounds and the mixtures are capable of reducing SOD activity versus the glutamate-induced damage (p<0.05); in particular, MIX 2 succeeds in reducing the damage in a more effective and synergistic manner compared to the respective single active compounds (p<0.05) (Table 31). All agents show to be effective irrespective of the time at which Riluzole is administered. Furthermore, if Riluzole is administered before or simultaneously, the effect of MIX 2 is amplified compared to MIX 1 (p<0.05).

[0328] Table 31 : SOD activity given by the components and mixtures in combination with Riluzole

[0329] 31.1 Pre-Riluzole 31.2 Co-Riluzole

[0330] 31.3 Post-Riluzole Conclusions

[0331] Mix 1 and Mix 2 were both demonstrated to be effective in counteracting motor neuron neurodegeneration.

[0332] The mixtures have comparable activity on most of the markers, but in some cases Mix 2 shows to be statistically superior to Mix 1.

[0333] The data demonstrate that it is possible to combine both mixtures with treatment with the drug of first choice, Riluzole, because in all cases the biological effectiveness of the drug is enhanced by the combination with the two formulations.

Claims

CLAIMS1. A mixture comprising or, alternatively, consisting of: i) an extract of chamomile; ii) (ii) an extract of turmeric; iii) (ill) L-acetyl carnitine or a salt thereof; iv) (iv) a blueberry extract; and v) Lipoic acid or a salt thereof; and / or a bergamot extract.

2. The mixture according to claim 1 , wherein said I) chamomile extract is preferably a dry extract of Matricaria chamomilla titrated in apigenin, with a minimum apigenin titre > 2% w / w.

3. The mixture according to claim 1 or 2, wherein said I) chamomile extract has an apigenin titre preferably comprised from 5% to 10% by weight, relative to the total weight of the dry extract.

4. The mixture according to any one of the preceding claims, wherein said ii) turmeric extract is preferably a dry extract of Curcuma longa titrated in curcuminodes, with a minimum curcuminoid titre > 30% w / w.

5. The mixture according to any one of the preceding claims, wherein said ii) turmeric extract has a curcuminoid titre comprised from 30% to 50% by weight, more preferably comprised from 35% to 45% by weight, relative to the total weight of the dry extract.

6. The mixture according to any one of the preceding claims, wherein said iii) L-acetyl carnitine is in salt form, preferably L-acetyl carnitine hydrochloride.

7. The mixture according to any one of the preceding claims, wherein said (iv) blueberry extract is preferably a dry extract of Vaccinium myrtillus titrated in anthocyanins, with a minimum anthocyanin titre > 20% w / w.

8. The mixture according to any one of the preceding claims, wherein said iv) blueberry extract has an anthocyanin titre comprised from 21% to 40%, more preferably comprised from 25% to 30% by weight, relative to the total weight of the dry extract.

9. The mixture according to any one of the preceding claims, wherein said v) is preferably bergamot extract.

10. The mixture according to any one of the preceding claims, wherein said bergamot extract v) comprises a total flavonoid content comprised from 20% to 50%, preferably comprised from 35% to 45%, more preferably from 38% to 40% by weight, relative to the total weight of the extract, wherein said flavonoids comprise naringin, neohesperidin, neoeriocitrin, melitidin, brutieridin.11 . The mixture according to claim 10, wherein said bergamot extract v) comprises naringin in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, e.g. 28 or 29%%, by weight, relative to the weight of total flavonoids, and a neohesperidin in an amount by weight comprised from 6% to 40%, preferably comprised from 15% to 35%, more preferably comprised from 20% to 30%, for example 29% or 30% by weight, relative to the weight of total flavonoids.

12. The mixture according to any one of the preceding claims, wherein said mixture comprises or, alternatively, consists of:i) an extract of chamomile; ii) an extract of turmeric; iii) L-acetyl carnitine or a salt thereof; iv) a blueberry extract; and v) a bergamot extract.

13. The mixture according to any one of the preceding claims, further comprising at least one additional ingredient (a), selected from the group comprising or, alternatively, consisting of:- a.i) at least one vitamin selected from the group comprising or, alternatively, consisting of vitamin C, vitamin E, vitamin B1 , vitamin B2, vitamin B6 and vitamin B12 or mixtures thereof,- a.ii) L-tryptophan,- a.iii) an extract of strawberry Fragaria,- a.iv) an extract of Crocus sativus, or mixtures thereof (a.i)+a.ii)+a.iii)+a.iv).

14. A composition comprising a mixture according to any one of claims 1 -13 and at least one acceptable pharmaceutical or food grade additive and / or excipient.

15. The composition according to claim 14, wherein said composition comprises or, alternatively, consists of: from 100 to 400 mg, preferably from 150 to 350 mg, even more preferably from 175 to 200 mg of a dry extract of Matricaria chamomilla titrated in apigenin (minimum titre > 2% w / w; the apigenin titre preferably being in a range of from 5% to 10%, more preferably 35% by weight relative to the total weight of the dry extract); from 30 to 200 mg, preferably from 50 to 160 mg, even more preferably from 60 to 100 mg of a dry extract of Curcuma longa titrated in curcuminoids (minimum titre > 30% w / w; the curcuminoid titre preferably being in a range of from 30% to 50%, preferably from 35% to 45%, more preferably 35% by weight relative to the total weight of the dry extract); from 200 to 2,000 mg, preferably from 300 to 1 ,000 mg, more preferably from 400 to 500 mg of L-acetyl carnitine (L-carnitine and / or carnitine) or a pharmaceutically acceptable salt thereof and / or mixtures thereof; from 50 to 200 mg, preferably from 60 to 180 mg, even more preferably from 80 to 120 mg of a dry extract of Vaccinium myrtillus titrated in anthocyanins (minimum titre > 20% w / w; the anthocyanin titre preferably being in a range of from 21 % to 40%, preferably from 25% to 30% by weight relative to the total weight of the dry extract); from 100 to 1 ,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of a lipoic acid or a pharmaceutically acceptable salt or mixtures thereof; and / or from 10 to 1 ,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of Bergamot extract (wherein said Bergamot extract preferably has a total flavonoid titre (neoeriocitrin, naringin, neohesperidin, melitidin, brutieridin) comprised from 20% to 50%, preferably from 25% to 45%, more from 30% to 40% w / w), and at least one acceptable pharmaceutical or food grade additive and / or excipient.

16. The composition according to claim 14, wherein said composition comprises or, alternatively, consists preferably of:from 100 to 400 mg, preferably from 150 to 350 mg, even more preferably from 175 to 200 mg, more preferably of 175 mg of a Matricaria chamomilla dry extract titrated in apigenin (minimum titre > 2 % w / w; the apigenin titre preferably being in a range of from 5% to 10%, more preferably 5% by weight relative to the total weight of the dry extract); from 30 to 200 mg, preferably from 50 to 160 mg, even more preferably from 60 to 100 mg of a dry extract of Curcuma longa titrated in curcuminoids (minimum titre > 30% w / w; the titre in curcuminoids preferably being in a range of from 30% to 50%, preferably from 35% to 45%, more preferably 35% by weight relative to the total weight of the dry extract); from 200 to 2,000 mg, preferably from 300 to 1 ,000 mg, more preferably from 400 to 500 mg of L-acetyl carnitine (L-carnitine and or carnitine) or a pharmaceutically acceptable salt thereof and / or mixtures thereof; from 50 to 200 mg, preferably from 60 to 180 mg, even more preferably from 80 to 120 mg of a dry extract of Vaccinium myrtillus titrated in anthocyanins (minimum titre > 20% w / w, the anthocyanin titre preferably being in a range of from 21 % to 40%, preferably from 25% to 30% by weight relative to the total weight of the dry extract); from 10 to 1 ,000 mg, preferably from 200 to 600 mg, even more preferably from 300 to 400 mg of Bergamot extract (wherein said Bergamot extract preferably has a total flavonoid titre (neoeriocitrin, naringin, neohesperidin, melitidin, brutieridin) comprised from 20% to 50%, preferably from 25% to 45%, more from 30% to 40% w / w), and at least one acceptable pharmaceutical or food grade additive and / or excipient.

17. The mixture according to any one of claims 1 -13, for use as a medicament.

18. The composition according to any one of claims 14-16, for use as a medicament.

19. The mixture or the composition for use according to claim 17 or 18, for the protection of central nervous system function and / or to counteract motor neuron degeneration and demyelination processes and / or in a method of preventing and / or treating neurodegenerative diseases affecting the motor neuron and / or due to demyelination processes, such as Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS).

20. The mixture or composition for use according to claim 19, wherein said mixture and / or said composition is more preferably for use in a method of preventing and / or treating Amyotrophic Lateral Sclerosis (ALS).

21. The mixture or composition for use according to claim 20, for use in combination with at least one drug conventionally used for treating Amyotrophic Lateral Sclerosis (ALS), wherein said at least one drug is Riluzole.

22. The mixture or composition for use according to any one of claims 17-21 , for oral use, preferably in the form of a capsule, tablet, sachet, stick, orosoluble stick, oral gel, or granules.

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