WO2019082136A1

WO2019082136A1 - A composition for the treatment and/or prevention of neurodegenerative diseases

Info

Publication number: WO2019082136A1
Application number: PCT/IB2018/058366
Authority: WO
Inventors: Umberto DI MAIO
Original assignee: Neilos S.r.l.
Priority date: 2017-10-26
Filing date: 2018-10-26
Publication date: 2019-05-02
Also published as: EP3700631A1; IT201700121764A1

Abstract

The invention relates to a synergistic composition of naturally occurring substances, which is particularly effective in the treatment and prevention of neurodegenerative diseases. The composition of the invention comprises the synergistic combination of phosphatidylserine, at least one tanshinone, and citicoline. The synergistic composition of the present invention may be provided in the form of a pharmaceutical or dietary supplement composition.

Description

A composition for the treatment and/or prevention of neurodegenerative diseases

The present invention relates to a composition of substances preferably obtained from natural sources, which is effective in the prevention and/or treatment of neurodegenerative diseases.

Neurodegenerative diseases are characterised by the progressive loss of neurons from specific brain regions. These diseases include Parkinson's disease, Huntington's disease, Alzheimer's disease. Amyotrophic Lateral Sclerosis, as well as milder but still disabling conditions such as senile dementia.

In the case of Parkinson's disease and Huntington's disease, the loss of neurons that make up the basal ganglia lead to abnormalities in the management of movements. Instead, Alzheimer's disease is characterised by loss of neurons in the hippocampus and the cortex, leading to dysfunctions in memory and cognitive processes. Amyotrophic Lateral Sclerosis (ALS) is a disease in which muscle weakness results from the degeneration of spinal, bulbar and cortical motor neurons.

Overall, these diseases are quite widespread and represent a major burden for society. They are generally found in the advanced stages of life, in neurologically normal individuals, but some cases of disease development have been found during childhood.

Parkinson's disease is found in more than 1 % of individuals over the age of 65, whereas the prevalence of Alzheimer's disease is higher and accounts for 10% of the population. Huntington's disease is an inherited disorder with an autosomal dominant trait, whose incidence is decidedly lower than that of other neurodegenerative diseases, but over 50% of individuals in families carrying the gene are affected. ALS is also very rare, but it quickly leads to disability and death.

One of the peculiar characteristics of these diseases is the selectivity of the pathogenesis process for specific types of neurons. For example, in Parkinson's disease, extensive destruction of the substantia nigra dopaminergic neurons occurs, whereas neurons in the cortex and other areas of the brain are not affected. The neuronal damage induced by Alzheimer's disease affects the hippocampus and neocortex neurons more seriously and even within the cerebral cortex there is no uniformity in the areas of affected neurons. Even more impressive is the finding that in Huntington's disease the mutant gene responsible for this disorder is expressed throughout the brain and in many other organs, but pathophysiological changes are mostly observed in the neostriate.

The diversity in the pathogenesis of these neurodegenerative diseases has led those of skill to suggest the pathogenesis mechanism as a combination of genetic and environmental factors that influence intrinsic physiological characteristics and affect neuronal populations. Intrinsic factors may include susceptibility to damage induced by excitotoxicity, regional variations of capacity for oxidative metabolism, and production of toxic free radicals as by-products of cell metabolism.

For a long time, genetic predisposition was thought to play a key role in the etiopathogenesis of neurodegenerative diseases and several possible mechanisms have been identified. For example, Huntington's disease is transmitted by inheritance and the responsible gene defect has been identified. In Parkinson 's disease, instead, mutations of four different proteins - a-synuclein, parkin, UCHL 1 , and DJ- 1 - can lead to genetically determined disease forms. In Alzheimer's disease, mutations in genes encoding the precursor of the amyloid protein (APP) and proteins known as presenilins, probably involved in APP modification, lead to hereditary forms of this disease. However, the genetic origin of these diseases is rather rare.

Surely major etiological agents are environmental factors: infectious agents, toxins and brain damage can contribute to the pathogenesis process. These factors are more significant in Parkinson's disease, which can be caused by encephalitis or toxins such as "synthetic heroin", MPTP (N-methyl-4-phenyl- l ,2,3,6-tetrahydropyridine) or rotenone, which are able to damage dopaminergic neurons and lead to a condition very similar to Parkinson's disease. Instead, traumatic brain damage may lead to the onset of Alzheimer's disease.

Excitotoxicity is a phenomenon resulting from excessive glutamate concentration in the brain. Although glutamate is essential for normal brain function, its excessive concentration may lead to neuronal death induced by excitotoxicity. The destructive effects of glutamate are mediated by glutamate receptors, in particular NMDA (N-methyl-D- aspartate) receptors: the activation of this channel receptor by glutamate leads to the entrance of Ca²⁺ ions, which at high concentrations are able to induce several biological processes with destructive effects on neuronal cells.

Oxidative stress also plays a key role in the pathogenesis process. The excessive production of reactive compounds such as hydrogen peroxide and reactive oxygen species can lead to DNA damage, peroxidation of membrane lipids and neuronal death.

A Izhei er 's disease

Alzheimer's Disease (AD) causes progressive decline in cognitive abilities. A short-term memory dysfunction is usually the first clinical sign, whereas the long-term memory recall is relatively well preserved during disease progression. In the most advanced stages of this disease, several other cognitive abilities cease, such as calculating, exercising visuospatial skills and the ability to use objects and tools (ideomotor apraxia). The patient's level of excitation and state of alert are not impaired until the most advanced stages of the disease. Death, most often resulting from immobility complications such as pneumonia or pulmonary embolism, generally occurs 6- 12 years after the onset of the disease. Alzheimer's disease diagnosis is based on thorough clinical evaluations of the patient and appropriate laboratory tests, so that other AD-like disorders are ruled out. Alzheimer's disease is characterised by marked atrophy of the cerebral cortex and loss of cortical and subcortical neurons. AD's pathological characteristics are senile plaques, which are spherical accumulations of the β-amyloid protein accompanied by neuronal degenerative processes and formation of neurofibrillary tangles. The amount of senile plaques and neurofibrillary tangles is directly proportional to the cognitive deficits associated with this disease, preferably accumulating in the hippocampus and associative areas of the cortex, whereas areas such as the visual and the motor cortices are relatively spared. The neurochemical disorders that arise in AD have been extensively studied. Direct analysis of the content of neurotransmitters in the brain cortex shows a reduction of several neurotransmitters in parallel with neuronal loss. In particular, there is a marked and abnonnal reduction in acetylcholine content. The anatomical bases of cholinergic deficit are atrophy and degeneration of cholinergic neurons, particularly in the nucleus basalis, from where the cholinergic neurons of the entire cortex are innervated. Selective acetylcholine deficit in AD, as well as the observation of the dementia-like condition induced by cholinergic antagonists, led to the "cholinergic hypothesis" to explain the pathogenesis process of Alzheimer's disease.

The presence of β-amyloid aggregates is a peculiar feature of AD. Until recently, it was not yet entirely clear whether this was a process that could cause the disease or an effect of the latter, a consequence of neuronal death. The application of molecular genetics has helped to shed light on this issue, with the cloning of the β-amyloid precursor protein. The function of this protein is not known, but it is believed that it could act as a receptor for a hypothetical still unknown endogenous ligand. β-amyloid production appears to result from abnonnal proteolytic cleavage of APP by β-secretase (BACE, beta-site amyloid precursor protein cleaving enzyme 1 ). The main phamiacological approach to the treatment of Alzheimer's disease is the attempt to restore the cholinergic function of the brain. An initial approach consisted in the administration of acetylcholine precursors, such as choline chloride and phosphatidylcholine. Although these supplements are generally well tolerated, there is no significant evidence of the efficacy of these compounds from randomized clinical trials. A more promising strategy is the inhibition of acetylcholinesterase (AChE), an acetylcholine catabolic enzyme.

Physostigmine has a modest effectiveness in improving memory in animal learning models. However, the short half-life and the side effects arising from the cholinergic activation strongly limit its use in humans.

Tacrine showed a slight memory-enhancing effect when used in combination with choline. The strong side effects - abdominal cramps, anorexia, nausea, vomiting and diarrhea - which are observed in more than one third of patients treated with this drug, as well as increased serum transaminases, observed in more than 50% of treated patients, limit its use.

Donepezil is a selective AChE inhibitor in the central nervous system, with minor effects on peripheral tissues. The effectiveness in the treatment of patients with Alzheimer's disease is very low. Rivastigmine and galantamine produce a similar effect in reducing Alzheimer's disease symptoms. The side effects are less than those of tacrine and consist of nausea, diarrhea, vomiting and insomnia.

Other pharmacological treatments consist in the use of NMDA glutamate receptor antagonists, such as memantine. It is not known whether the mechanism of this dmg can act on the cause of the disease. Side effects of memantine consist of headache and dizziness.

Senile dementia

Historically, the term "dementia" indicated an irreversible, acquired and global deterioration of mental functions. The disorder is progressive and believed to be due to brain damage. Dementia can be classified as follows:

Dementia due to cerebrovascular disease. It can result from cerebral and peripheral vascular disorders. Neuropsychiatric symptoms can be caused by hypertensive encephalopathy, cerebral embolism or hemorrhage, thrombosis, and ischemia. In fact, cerebral ischemia is the main vascular cause of dementia. Recurrence of such events leads to the onset of multi-infarct dementia.

Dementia due to primary degenerative disorders. Primary degenerative disorders are seni le dementia, Alzheimer's disease, and some other presenile dementias. Senile dementia is a type of dementia that occurs in old age and is diagnosed in the absence of other possible causes of this disorder. Actually, there is more than one type of senile dementia: the various senile dementias are part of an extremely heterogeneous group, which includes more or less reversible forms of this disorder.

As in the case of Alzheimer's disease, the morphological changes in senile dementia are mainly observed in the neocortex, especially in the temporal, parietal and occipital areas, as well as in the grey matter. There is no epidemiological data available related to senile dementia. However, some interesting conclusions can be drawn from the numerous statistical studies concerning the broader category of dementias. According to these studies, dating back to 201 0, people with dementia around the world are approximately 36 million, divided as follows:

3% are aged between 65 and 74, 19% are aged between 75 and 84, and more than half are aged 85 and over.

6.8 million live in the United States. More than half of these people with dementia are over eighty years old.

750,000-800,000 live in the United Kingdom.

50-70% suffer from Alzheimer's disease, 25% from vascular dementia, 15% from Lewy body dementia, and the remaining percentage is affected by other existing forms of dementia.

The main, currently available pharmacological treatments for dementia are as follows:

Neuroleptic agents (typical or atypical antipsychotics);

- Antidepressant drugs;

Anti-dementia drugs (acetylcholinesterase inhibitors; nootropic agents);

Sedative/hypnotic drugs;

Anticonvulsants. However, the current pharmacological treatments have numerous side effects such as extrapyramidal reactions, tardive dyskinesia, parkinsonism, ataxia, cerebral edema, hypertension, hypotension, allergic reactions, hallucinations, gastrointestinal disorders, nausea and vomiting, rash, itching, headache, fatigue, pain.

Therefore, there is a need to provide alternative treatments to the existing ones, which are effective in the prevention and/or treatment of neurodegenerative diseases, but do not have the side effects and/or disadvantages of the state-of-the art treatments.

These and other needs are met by the present invention, which provides a composition characterised in that it comprises a synergistic combination of active ingredients obtained from natural sources, the aforesaid combination having proved particularly effective against neurodegenerative diseases.

The composition of the invention is as defined in appended claim 1. Further features and advantages of the invention are defined in the dependent claims. The claims form an integral part of the present specification.

A detailed description of some preferred embodiments of the invention is provided hereinafter.

The synergistic composition of the present invention is a supplement useful for the treatment and prevention of neurodegenerative diseases, more particularly Parkinson's disease, Huntington's disease, Alzheimer's disease, Amyotrophic Lateral Sclerosis, and senile dementia.

In the composition of the present invention the synergistic action takes place between phosphatidylserine, the at least one tanshinone and citicoline.

As will be explained in greater detail below, tanshinones are diterpenic compounds with a structure similar to that of abietane and are the main chemical compounds contained in the extract of plants of the Salvia genus, in particular in the extract of Salvia miltiorrhiza. The main bioactive compounds present in the extract of Salvia miltiorrhiza are tanshinone I (TNI), tanshinone IIA (T IIA), and cryptotanshinone (CPT). Phosphatidylserine is a molecule belonging to the family of membrane phospholipids, particularly abundant in the brain. Given its presence in the central nervous system, phosphatidylserine has been extensively studied for its potential neuroprotective activity. Several clinical studies in Europe and the United States have shown that phosphatidylserine extracted from the bovine cerebral cortex enhances the cognitive abilities in the elderly, including patients suffering from Alzheimer's disease and patients with age-related memory deficits.

Soybean-derived phosphatidylserine is decidedly cheaper than bovine phosphatidylserine, and above all is BSE (Bovine Spongiform Encephalopathy) risk-free. Although the acyl groups of soybean phosphatidylserine are slightly different from the bovine ones, in vivo studies have shown that the effects of the two types of phosphatidylserine are identical.

Several clinical studies have shown the effectiveness of phosphatidylserine in the treatment of cognitive disorders.

In a clinical trial performed in elderly patients with age-related cognitive disorders, 300 mg/day of phosphatidylserine were administered over a 3-month period. The trial showed that citicoline is able to improve the memory-related scores in the Wechsler test, especially in the test components assessing visual memory.

Another clinical study reported that the administration of phosphatidylserine over a 12- week period can improve mnemonic functions, such as remembering names and faces, in elderly people with age-related cognitive deficits.

A more recent study, in 2010, evaluated two dosages of citicoline: 100 mg and 300 mg/day. The subjects participating in the study were divided into 3 groups: the first was administered a placebo, the second 100 mg/day of phosphatidylcholine, and the third 300 mg/day of phosphatidylserine. The duration of the study was 6 months followed by a 3- month follow-up, during which the patients did not follow any treatment. The primary outcome was assessed with different tests and questionnaires on memory and cognitive functions ((HDS-R, MMSE, RBMT). In subjects with a low baseline score in these tests, a statistically significant change in these scores was observed in the phosphatidylserine groups.

Salvia miltio hiza, the botanical name for the plant known as red sage or Chinese sage, is widely used in traditional Chinese medicine for the treatment of cerebrovascular and cardiovascular disorders, as well as inflammatory diseases. Tanshinones, diterpenic compounds with a structure similar to that of abietane, are the main chemical compounds contained in the Salvia miltiorrhiza extract. The main bioactive compounds are tanshinone I (TN I), tanshinone II A (TNIIA), and cryptotanshinone (CPT). These compounds have attracted attention because of their different pharmacological effects, which include the anti-inflammatory and anti-tumour effects and the cerebrovascular protection activities. Tanshinone II A has potential effects against diabetes, neurodegenerative diseases and cardiac hypertrophy. In addition to the anti-inflammatory and anti-tumour effects, tanshinone I has the ability to enhance memory and learning and improve memory disorders.

Several studies were carried out to assess the activity of Salvia miltiorrhiza and tanshinones in the treatment of cardio- and cerebrovascular diseases such as myocardial infarction, atherosclerosis, hyperlipidemia, hypertension, and stroke. Tanshinone IIA and tanshinone IIA sodium sulfonate have shown effectiveness in reducing the extent of myocardial infarction and improving cardiac function, through the activation of phosphatidylinositol 3-kinase (PI3K). Other pharmacological effects consist in inhibiting platelet aggregation, inhibiting the formation of atherosclerotic lesions and hyperlipidemia, reducing lipoprotein oxidation and macrophage accumulation at the vascular tunica.

Further pharmacological activities of tanshinones consist in inhibiting tumour cell proliferation. The anti-tumour action mainly derives from the cytotoxicity of TNIIA, TNI and CPT, together with cell cycle arrest and apoptosis. The mechanism could consist in the up-regulation of pro-apoptotic proteins.

Tanshinone IIA has also shown an important anti-inflammatory activity, by inhibiting the expression of inflammatory mediators such as IL- Ι β, IL-6 and rumour necrosis factor a (TNF-α). In addition, TNIIA is capable of inhibiting lipopolysaccharide-induced NF-kB activation, through inhibition of the N I /Ι Κα (NF-KB-inducing kinase/ IkappaB alpha kinase), ERK- 1 /2, p38 and INK (c-Jun N-terminal kinase) pathways. The antiinflammatory effect may also partly result from activation of the TNF receptor-associated factor (TRAF) 2/3/6 and inhibition of the toll-like receptor (TLR) signalling pathway.

Tanshinones are known as natural antioxidants. The mechanism consists in the formation of a quinone adduct of the lipid radical to form a stabilized radical. TNIIA is capable of preventing DNA damage resulting from lipid peroxidation in liver cells, through the radical scavenging activity against free lipid radicals and the stopping of peroxidation chain reactions. Pre-incubation with TNIIA significantly reduces the death of ECV-304 cells and J774 macrophages in a dose-dependent manner.

The protein binding the cAMP response element (cAMP response element binding protein, CREB) is a ubiquitously expressed transcription factor, which is known to regulate the expression of genes that are important for the proliferation, differentiation, growth, survival and adaptation of all cell types. The phosphorylated (active) form of the CREB protein has been extensively investigated in ischemia models. Cerebral ischemia, in fact, triggers marked phosphorylation of CREB, DNA binding of CRE (cAMP responsive elements), and the transcription of several neuroprotective molecules, such as the anti- apoptotic protein Bcl-2 (B cell lymphoma/leukemia-2) and BDNF (brain-derived neurotrophic factor). BDNF promotes cell survival and reduces infarction volume after ischemia. In neurons, the CREB protein plays an important role in synaptic plasticity, neurogenesis, and axonal growth following ischemia. TORC proteins (TORC l , TORC2, TORC3) have recently been identified as potent and selective co-activators of the CREB protein and the CREB/CRE pathway. TORC proteins are generally found in the cytoplasm, but as a result of increased intracellular Ca²⁺ or cAMP-mediated extracellular signals they translocate into the nucleus and enhance the transcription of CREB protein target genes. TORC l isoform is the most expressed in brain tissue, particularly in the hippocampus, cortex and cerebellum. Recent studies have shown that TORC l acts as an essential regulator for BDNF transcription in long-term enhancement of the hippocampus. TORC 1 accumulation in the nucleus can induce BDNF gene transcription in the hippocampus and the retinal ganglion cells following ischemic damage.

One study assessed the ability of Tanshinone 1IA to induce TORC 1 activation and increase gene expression of TORC 1 , pCREB and BDNF in the initial phase that follows the cerebral ischemia. Researchers detected dynamic expression of TORC 1 , pCREB and BDNF in the cerebral cortex. Gene expression of these proteins showed two peaks: pCREB and BDNF expression reached 2 maximum peaks at 3 and 72 hours, while TORC 1 expression reached the maximum value 3 and 48 hours after the ischemic event. Citicoline is the International non-proprietary name for cytidine 5'-diphosphocholine. This is a compound marketed in several countries, both as a drug and as a dietary supplement.

Citicoline is a complex organic molecule that acts as a biosynthetic intermediate in the synthesis of membrane phospholipids. Citicoline belongs to the nucleotide family and is composed of ribose, pyrophosphate, cytosine, and choline.

Choline is a quaternary ammonium salt present in three major metabolic pathways:

1 . Phospholipid synthesis through phosphorylcholine;

2. Acetylcholine synthesis;

3. Synthesis of betaine, a major donor of methyl groups.

Cytidine becomes one of the main components of nucleic acids after its conversion to cytidine triphosphate (CTP). In citicoline metabolic pathway, choline is phosphorylated by the enzyme choline kinase to form phosphorylcholine; subsequently, the latter combines with cytidine triphosphate to form citicoline. Citicoline reacts with diacylglycerol, forming phosphatidylcholine by means of choline phosphotransferase, which catalyzes this reaction.

Results obtained from investigations carried out with citicoline and isotopic markers, 3H- CDP-methyl- 14C-Cho, demonstrated that citicoline, after oral or intravenous administration, undergoes rapid hydrolysis to cytidine 5'-monophosphate and phosphocholine. The latter are further dephosphorylated by phosphatases, so that they can cross the blood-brain barrier. Exogenous citicoline, after oral administration, is rapidly hydrolysed to choline and cytidine, which are absorbed by the small intestine. These two molecules serve as endogenous citicoline precursors and contribute to increase the supply of this substance in the body.

Pharmacokinetic studies have shown that the citicoline plasma level curve has two absorption peaks: the first one hour after ingestion and the second about 24 hours later. Clearance is biphasic and mainly due to CO2 expiration and urinary excretion. Pulmonary clearance half-life is 56 hours and urinary clearance half-life is 71 hours.

Citicoline increases the formation of phosphatidylcholine and other phospholipids in the brain, as revealed by in vitro and in vivo studies, and at the same time prevents ischemia- induced accumulation of free fatty acids. These two effects should be anti-apoptotic and neuroprotective. The neuroprotective effect was observed in several experimental models of cerebral ischemia, stroke, hypoglycaemic hypoxia and neuronal apoptosis induced by deposition of β-amyloid precursor protein, and hypoperfusion. The neuroprotection that follows the administration of citicoline could be associated with stimulation of phosphatidylcholine synthesis or stimulation of S-adenosylmethionine synthesis, which stabilizes the membrane and prevents the release of arachidonic acid.

The neuroprotection mechanism may derive from the ability of citicoline to repair neuronal membranes, which derives from an increase in phosphatidylcholine synthesis, the repair of cholinergic neurons through the enhancement of acetylcholine production and the reduction of the concentration of free fatty acids.

A further mechanism that contributes to the neuroprotective action of citicoline is the prevention of excitotoxicity. In fact, citicoline administration has been shown to be able to prevent kainic acid-induced excitotoxicity. The mechanism is based on the prevention of nitrosative stress and inhibition of activation of kainic acid-induced ERK kinase.

In addition to neuroprotective properties, citicoline is known to increase the synthesis of acetylcholine, dopamine noradrenaline and serotonin in several brain areas. The cholinergic effect of citicoline is due to the fact that choline is the precursor of acetylcholine. The mechanism leading to increased catecholamine synthesis is less known.

Preclinical studies in animals have shown that citicoline can improve memory and learning. Memory disorders in the elderly can be due to different causes, such as the reduction of neurotransmitter formation, poor circulation (vascular dementia) or diseases such as Alzheimer's disease. The effectiveness of citicoline in the treatment of cognitive deficits found in old age would appear to depend on the latter cause. The potential of citicoline in the treatment of age-related memory disorders was demonstrated in a double-blind clinical study conducted in 84 elderly patients with mild or moderate memory loss. Patients were treated with 1000 mg/day of citicoline or placebo for a period of 6 weeks. The results showed an improvement in attention and memory. The results of a recent meta-analysis have shown that citicoline is moderately effective in cognitive and behavioural disorders associated with brain disorders in the elderly. Experimental evidence also demonstrates the effectiveness of this molecule in the treatment of damage caused by cerebral ischemia and head trauma, in Parkinson's disease and Alzheimer's disease.

It is interesting to note the low toxicity of citicoline, which has an LD50 of 4.6 g/kg in mice and 4. 1 5 g/kg in rats when administered intravenously, and 8 g/kg in both species when administered orally. In a phase IV clinical trial, about 3000 elderly patients with neurological disorders were treated with citicoline at a dose of 600 mg/day over, a period of 15-60 days: the incidence of side effects was 5.01 % and none of them was serious.

As indicated above, the composition of the present invention is effective in the treatment and/or prevention of neurodegenerative diseases, more particularly Parkinson's disease, Huntington's disease. Alzheimer's disease, Amyotrophic Lateral Sclerosis, and senile dementia.

The effectiveness of the composition derives from the following combined activities of its components:

• Neuroprotective effect;

• Anti-inflammatory effect;

• Antioxidant effect.

Tanshinones, for example present in the Salvia miltiorrhiza extract, have the ability to increase the activation of the TORC 1 protein, which in turn is involved in the phosphorylation of the CREB protein, an important transcription factor in gene expression of neuroprotective factors such as BDNF. Phosphatidylserine is one of the main phospholipids present in the neuronal membrane and an important structural element at the level of the central nervous system. Supplementation of this molecule can contribute to neuronal protection. Citicoline exerts its neuroprotective effect through different mechanisms: it is a source of choline in the central nervous system and contributes to the synthesis of phosphatidylcholine, inhibits the deposition of the β-amyloid protein, contributes to repairing the cholinergic neurons and increasing the production of acetylcholine, prevents excitotoxicity damage induced by glutamate.

The Salvia miltiorrhiza extract has an important anti-inflammatory effect, by inhibiting NF-kB, and consequently the gene expression of cytokines and pro-inflammatory enzymes.

Lastly, citicoline and the Salvia miltiorrhiza extract have an antioxidant and radical scavenger effect, which can further contribute to neuronal protection from oxidative stress damage. The combination of these mechanisms of action is useful for the prevention and/or treatment of neuronal degeneration and diseases resulting therefrom.

In a preferred embodiment, phosphatidylserine is administered in an amount comprised between 1 mg/day and 500 mg/day (e.g. 1 mg/day; 2 mg/day; 3 mg/day; 4 mg/day; 5 mg/day; 6 mg/day; 7 mg/day; 8 mg/day; 9 mg/day; 10 mg/day; 15 mg/day; 20 mg/day; 25 mg/day; 30 mg/day; 35 mg/day; 40 mg/day; 45 mg/day; 50 mg/day; 55 mg/day; 60 mg/day; 65 mg/day; 70 mg/day; 75 mg/day; 80 mg/day; 85 mg/day; 90 mg/day; 95 mg/day; 100 mg/day; 1 10 mg/day; 120 mg/day; 130 mg/day; 140 mg/day; 150 mg/day;

160 mg/day; 170 mg/day; 180 mg/day; 190 mg/day; 200 mg/day; 210 mg/day; 220 mg/day; 230 mg/day; 240 mg/day; 250 mg/day; 260 mg/day; 270 mg/day; 280 mg/day;

290 mg/day; 300 mg/day; 310 mg/day; 320 mg/day; 330 mg/day; 340 mg/day; 350 mg/day; 360 mg/day; 370 mg/day; 380 mg/day; 390 mg/day; 400 mg/day; 410 mg/day;

420 mg/day; 430 mg/day; 440 mg/day; 450 mg/day; 460 mg/day; 470 mg/day; 480 mg/day; 490 mg/day; 500 mg/day), for example by administration of a dosage form wherein phosphatidylserine is present in the composition in an amount comprised between

0.01% and 50% by weight based on the total weight of the composition, preferably in an amount of between 0.1% and 40 % by weight based on the total weight of the composition.

For example, phosphatidylserine is present in the composition in an amount of 0.01%;

0.02%; 0.03%; 0.04%; 0.05%; 0.06%; 0.07%; 0.08%; 0.09%; 0.1 %; 0.2%; 0.3%; 0.4%;

0.5%; 0.6%; 0.7%; 0.8%; 0.9%; 1 %; 2%; 3%; 4%; 5%; 6%; 7%; 8%; 9%; 10%; 1 1%;

1 2%; 13%; 14%; 15%; 16%; 17%; 18%; 19%; 20%; 21%; 22%; 23%; 24%; 25%; 26%; 27%; 28%; 29%; 30%; 31 %; 32%; 33%; 34%; 35%; 36%; 37%; 38%; 39%; 40%; 41 %;

42%; 43%; 44%; 45%; 46%; 47%; 48%; 49%; 50% by weight based on the total weight of the composition.

The aforesaid values (mg/day and %) can be combined in any range.

In another preferred embodiment, tanshinone/tanshinones is/are present in the composition in the fonn of an extract of a plant of the genus Salvia, preferably Salvia miltiorrhiza. In this embodiment, the Salvia miltiorrhiza extract, preferably titrated in tanshinones, is administered in an amount comprised between 1 mg/day and 1000 mg/day (e.g. 1 mg/day; 2 mg/day; 3 mg/day; 4 mg/day; 5 mg/day; 6 mg/day; 7 mg/day; 8 mg/day; 9 mg/day; 10 mg/day; 1 5 mg/day; 20 mg/day; 25 mg/day; 30 mg/day; 35 mg/day; 40 mg/day; 45 mg/day; 50 mg/day; 55 mg/day; 60 mg/day; 65 mg/day; 70 mg/day; 75 mg/day; 80 mg/day; 85 mg/day; 90 mg/day; 95 mg/day; 100 mg/day; 1 10 mg/day; 120 mg/day; 130 mg/day; 140 mg/day; 150 mg/day; 160 mg/day; 170 mg/day; 180 mg/day; 190 mg/day; 200 mg/day; 210 mg/day; 220 mg/day; 230 mg/day; 240 mg/day; 250 mg/day; 260 mg/day; 270 mg/day; 280 mg/day; 290 mg/day; 300 mg/day; 310 mg/day; 320 mg/day; 330 mg/day; 340 mg/day; 350 mg/day; 360 mg/day; 370 mg/day; 380 mg/day; 390 mg/day; 400 mg/day; 410 mg/day; 420 mg/day; 430 mg/day; 440 mg/day; 450 mg/day; 460 mg/day; 470 mg/day; 480 mg/day; 490 mg/day; 500 mg/day; 550 mg/day; 600 mg/day; 650 mg/day; 700 mg/day; 750 mg/day; 800 mg/day; 850 mg/day; 900 mg/day; 950 mg/day; 1000 mg/day), for example by administration of a dosage form wherein the extract is present at a concentration comprised between 0.01 % and 60% by weight based on the total weight of the composition, preferably in an amount comprised between 1% and 50% by weight based on the total weight of the composition. For example, the extract is present in the composition at a concentration of 0.01%; 0.02%; 0.03%; 0.04%; 0.05%; 0.06%; 0.07%; 0.08%; 0.09%; 0.1%; 0.2%; 0.3%; 0.4%; 0.5%; 0.6%; 0.7%; 0.8%; 0.9%; 1 %; 2%; 3%; 4%; 5%; 6%; 7%; 8%; 9%; 10%; 1 1%; 12%; 13%; 14%; 15%; 16%; 17%; 1 8%; 19%; 20%; 21 %; 22%; 23%; 24%; 25%; 26%; 27%; 28%; 29%; 30%; 31%; 32%; 33%; 34%; 35%; 36%; 37%; 38%; 39%; 40%; 41 %; 42%; 43%; 44%; 45%; 46%; 47%; 48%; 49%; 50%; 51 %; 52%; 53%; 54%; 55%; 56%; 57%; 58%; 59%; 60% by weight based on the total weight of the composition.

The aforesaid values (mg/day and %) can be combined in any range.

In yet another preferred embodiment, citicoline is administered in an amount comprised between 50 mg/day and 5000 mg/day (e.g. 50 mg/day; 55 mg/day; 60 mg/day; 65 mg/day; 70 mg/day; 75 mg/day; 80 mg/day; 85 mg/day; 90 mg/day; 95 mg/day;- 100 mg/day; 1 10 mg/day; 120 mg/day; 130 mg/day; 140 mg/day; 150 mg/day; 160 mg/day; 170 mg/day; 180 mg/day; 190 mg/day; 200 mg/day; 210 mg/day; 220 mg/day; 230 mg/day; 240 mg/day; 250 mg/day; 260 mg/day; 270 mg/day; 280 mg/day; 290 mg/day; 300 mg/day; 310 mg/day; 320 mg/day; 330 mg/day; 340 mg/day; 350 mg/day; 360 mg/day; 370 mg/day; 380 mg/day; 390 mg/day; 400 mg/day; 410 mg/day; 420 mg/day; 430 mg/day; 440 mg/day; 450 mg/day; 460 mg/day; 470 mg/day; 480 mg/day; 490 mg/day; 500 mg/day; 550 mg/day; 600 mg/day; 650 mg/day; 700 mg/day; 750 mg/day; 800 mg/day; 850 mg/day; 900 mg/day; 950 mg/day; 1000 mg/day; 1 100 mg/day; 1200 mg/day; 1300 mg/day; 1400 mg/day; 1500 mg/day; 1600 mg/day; 1700 mg/day; 1800 mg/day; 1900 mg/day; 2000 mg/day; 2100 mg/day; 2200 mg/day; 2300 mg/day; 2400 mg/day; 2500 mg/day; 2600 mg/day; 2700 mg/day; 2800 mg/day; 2900 mg/day; 3000 mg/day; 3100 mg/day; 3200 mg/day; 3300 mg/day; 3400 mg/day; 3500 mg/day; 3600 mg/day; 3700 mg/day; 3800 mg/day; 3900 mg/day; 4000 mg/day; 4100 mg/day; 4200 mg/day; 4300 mg/day; 4400 mg/day; 4500 mg/day; 4600 mg/day; 4700 mg/day; 4800 mg/day; 4900 mg/day; 5000 mg/day), for example by administration of a dosage form wherein citicoline is present at a concentration comprised between 1% and 95% by weight based on the total weight of the composition, preferably in an amount comprised between 10% and 90% by weight based on the total weight of the composition. For example, citicoline is present in the composition at a concentration of 1 %; 2%; 3%; 4%; 5%; 6%; 7%; 8%; 9%; 10%; 1 1 %; 12%; 13%; 14%; 15%; 16%; 1 7%; 18%; 19%; 20%; 21 %; 22%; 23%; 24%; 25%; 26%; 27%; 28%; 29%; 30%; 3 1 %; 32%; 33%; 34%; 35%; 36%; 37%; 38%; 39%; 40%; 41%; 42%; 43%; 44%; 45%; 46%; 47%; 48%; 49%; 50%; 51%; 52%; 53%; 54%; 55%; 56%; 57%; 58%; 59%; 60%; 61 %: 62%; 63%; 64%; 65%; 66%; 67%; 68%; 69%; 70%; 71%; 72%; 73%; 74%; 75%; 76%; 77%; 78%; 79%; 80%; 81 %; 82%; 83%; 84%; 85%; 86%; 87%; 88%; 89%; 90%; 91 %; 92%; 93%; 94%; 95% by weight based on the total weight of the composition.

The aforesaid values (mg/day and %) can be combined in any range.

The dosage form can be a pharmaceutical composition or a supplement, preferably an oral pharmaceutical form, such as for example a tablet, a capsule or a sachet, including the above-mentioned active ingredients mixed together. Specific examples of combinations of active ingredients according to the invention are:

- A tablet containing, as active ingredients, 500 mg citicoline, 100 mg sage titrated in tanshinones, and 20 mg phosphatidylserine;

- A sachet containing, as active ingredients, 1000 mg citicoline, 200 mg sage titrated in tanshinones, and 40 mg phosphatidylserine.

The following examples are provided for illustration purposes only and do not limit the scope of the invention as defined in the appended claims. EXAMPLES

Some formulation examples are reported, with the quantities of the related active substances occurring in each dosage unit.

EXAMPLE 1 : Pharmaceutical Form: 1030 mg tablet

EXAMPLE 2: Phannaceutical Fonn: 1300 mg tablet

EXAMPLE 3 : Phannaceutical Fonn: 1500 mg sachet

Active ingredient Daily Dose

Citicoline 500 mg

Phosphatidylserine 300 mg

Salvia miltiorrhiza, d.e. 200 mg

EXAMPLE 4: 1950 mg sachet

Active ingredient Daily Dose

Citicoline 1000 mg

Phosphatidylserine 300 mg

Salvia miltiorrhiza, d.e. 200 mg

EXAMPLE 5: Pharmaceutical Fonn: 3250 mg sachet

Active ingredient Daily Dose

Citicoline 2000 mg

Phosphatidylserine 300 mg

Salvia miltioiThiza, d.e. 200 mg EXAMPLE 6: Pharmaceutical Form: 975 mg capsule

EXAMPLE 8

The composition of Example 7 was prepared by mixing citicoiine, phosphatidylserine and the Salvia miltiorrhiza extract with the excipients and then compressing the above into the tablet form. The tablet was then coated with a film based on hydroxypropyl cellulose and titanium dioxide.

Claims

1. A pharmaceutical or dietary supplement composition comprising, as active substances, phosphatidylserine, at least one tanshinone, and citicoline.

2. The pharmaceutical or dietary supplement composition according to claim 1 , wherein the at least one tanshinone is selected from the group consisting of tanshinone I (TNI), tanshinone I1A (TNI1A), cryptotanshinone (CPT), and combinations thereof.

3. The pharmaceutical or dietary supplement composition according to claim 1 or 2, wherein the at least one tanshinone is present in the composition in the form of an extract of a plant of the Salvia genus.

4. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 3, wherein the plant of the Salvia genus is Salvia miltiorrhiza.

5. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 4, wherein phosphatidylserine is present in the composition in an amount comprised between 0.01 % and 50% by weight based on the total weight of the composition, preferably in an amount comprised between 0. 1 % and 40 % by weight based on the total weight of the composition.

6. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 5, wherein the at least one tanshinone is present in the composition in the form of an extract of the Salvia genus, which is present in an amount comprised between 0.01 % and 60% by weight based on the total weight of the composition, preferably in an amount comprised between 1 % and 50% by weight based on the total weight of the composition.

7. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 6, wherein citicoline is present in the composition in an amount comprised between 1 % and 95% by weight based on the total weight of the composition, preferably in an amount comprised between 10% and 90% by weight based on the total weight of the composition.

8. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 7, which is formulated as an oral dosage form.

9. The pharmaceutical or dietary supplement composition according to any one of claims 1 to 8, for use in the prevention and/or treatment of neurodegenerative diseases.

10. The pharmaceutical or dietary supplement composition for use according to claim 9, wherein the neurodegenerative disease is Parkinson's disease, Huntington's disease,

Alzheimer's disease. Amyotrophic Lateral Sclerosis or senile dementia.