DESCRIPTION

Field of application

The present invention relates to the field of pharmaceutical and food supplement industry.

In particular, the invention relates to a natural base composition for mood regulation, the prevention and/or treatment of mood and/or anxiety disorders.

Prior art

The term “mood disorder” refers to a large class of mood alteration symptoms of the individual, as well as to maladaptation to relational and environmental life conditions, influencing the interpersonal and/or working sphere.

Generally, these swings occur suddenly, are of excessive degree and appear with a series of symptoms which tend to become chronic and to considerably compromise a person’s everyday life.

Depression is among the most recurring mood disorders, namely a form of mental suffering characterized by lowering of mood.

In particular, the“unipolar depression” is associated with events wherein a high emotional stress is involved and is typically accompanied by anxiety and agitation symptoms.

Depression may occur in different ways assuming different aspects in relation to such factors as age, personality and causes. There is a considerable mental and motor involvement in the most serious forms up to the melancholic stupor with mutism and immobility. More rarely, states of restlessness and excitement based on a considerable anxious component may be observed. When depression is associated with irritability, bad mood and aggression, such a condition is known as “dysphoria”.

It is known that the depressive state comprises mental, psychomotor and somatic manifestations. This results in a condition of suffering and pain, which appears with sadness, pessimism, slowing down of ideation and motor activity, reduction in self-evaluation and self-confidence, loss of interests, difficulty in performing the usual activities, tendency to cry.

In particular, it is known that other diseases can arise from a depressive condition, among which anxiety and recurring sleep disorders, which can worsen and intensify the symptoms connected to depression.

Specifically, anxiety, namely a mental state characterized by a sensation of fear which is not connected to any specific stimulus, is often accompanied by different physical conditions, such, for example, palpitations, chest pains, shortness of breath, nausea and tremor.

From the emotional point of view, anxiety causes a sense of terror or panic, nausea and shivers. From the behavioural point of view both voluntary and involuntary behaviours may occur, aimed at escaping or avoiding the source of anxiety; these behaviours are recurring and often not adaptive, since they are extreme in anxiety disorders.

For example generalized anxiety manifestations, such as a state of excessive anxiety without any manifest or clear reasons, may be distinguished in case of pathological anxiety disorder; panic attacks, such as sudden unbearable fear attacks, appear along with pronounced somatic symptoms, such as sweating, tachycardia, chest pain, tremor, suffocating sensation; phobias, namely strong fears of objects and specific situations; obsessive-compulsive disorders, namely ritual compulsive behaviours caused by irrational anxiety.

As for sleep disorders, it is known that, typically, such disorders usually lead to the development of insomnia, characterized by a difficulty in falling asleep and sleep continuity and in general by a not refreshing sleep.

Insomnia may cause discomfort also during the day; in fact, patients often experience attention difficulties, poor concentration and memory deficiency, irritability, energy reduction, daytime sleepiness, generic fatigue sensation and discomfort.

Anxiety and sleeping disorders, as well as other diseases caused by depression, are typically alleviated when the patient take drugs which directly act on the depressive state, the so-called “antidepressant medicines”.

Since it is known that different neurotransmitters of the central nervous system are involved in depression and it is also known that those involved in the antidepressant ways are noradrenaline and serotonin, typically antidepressant medicines especially act by precisely increasing the transmission of noradrenaline and serotonin.

The most common antidepressant drugs are represented by selective serotonin reuptake inhibitors, selective noradrenaline reuptake inhibitors, monoamine oxidase enzyme inhibitors (an enzyme which decomposes monoamines), non-selective monoamine reuptake inhibitors and non-selective receptor antagonists.

The beneficial effects of such drugs are various. In fact, in addition to the properly called antidepressant effect, namely a significant reduction of sad mood, pessimism, anguish and somatic symptoms, an effect that combats anxiety and agitation, namely the prevalent symptoms of such pathology, and a stimulating effect, which acts on apathy, can also be found.

However, the above-mentioned benefits are often accompanied by important side effects, among which sedation, postural hypotension, dry mouth, blurred vision, constipation, nausea, insomnia, weight gain, agitation.

Furthermore, such drugs may result to be dangerous if overdosed and, furthermore, they may interact with other drugs, such as anaesthetic, anti-hypertensive and non-steroidal anti-inflammatory drugs.

It is known in particular that these drugs are contraindicated in association with monoamine oxidase inhibitors because of the risk of a serotonergic reaction characterized by hyperthermia, muscle stiffness e/o cardiocirculatory collapse.

It is further known that the antidepressant drugs need at least two weeks to bring about some beneficial effects even if their pharmacological effects are immediately produced, which indicates that the adaptive secondary modifications play an important role.

A further relevant aspect regards the intake of drugs, for example sleep- inducing/ anxiolytic and antidepressant drugs, even if the pathological conditions of the patient do not necessarily require the use thereof. This may result in pharmacological dependency manifestations in such specific patients, more often due to sleep-inducing drugs.

Beside the above-mentioned known drugs, there is a growing interest for natural remedies which are able to alleviate the predominant depression symptoms.

For example, it is known that the consumption of the extract of the Hypericum officinal plant (Hypericum Perforatum) involves benefits for the reduction of anxiety, melancholy and sleep disorders. These properties are mainly ascribed to flavonoids, in particular to hypericin, contained in the extract of such a plant which carry out an antidepressant action in case of mood alterations.

It is further known that the griffonia ( Grifforda simplicifolia) seeds, a woody plant of tropical African areas, contain 5-hydroxy-tryptophan (5- HTP), a precursor of serotonin. Such seeds act therefore by directly increasing the serotonin levels in the organism, thus improving mood.

Patent application ES 2573542 describes a powder composition comprising the saffron stigma extract for therapeutic use, in particular for the treatment of mood disorders related to depression.

Patent application US 2005/0208156 describes the use of different compounds, such as saffron, for the treatment of schizophrenia; saffron can be used both fresh and in the dried form, all parts or only some thereof depending on the plant growth stage may be used and preferably the stigma extract is used.

US 2017/239312 describes a weight loss system , which comprises a weight loss accelerator formulation, comprising different active ingredients, such as saffron, and a weight loss control formulation , comprising, among other active ingredients , green tea and L-theanine extract.

US 201 1/014277 describes a composition for tobacco smoking cessation , comprising, among other active ingredients, magnolia extract, green tea extract, L-theanine and melatonin.

US 2006/018975 describes a food supplement composition for the treatment of mood disorders, comprising, among other active ingredients, magnolia bark extract, green tea extract and L-theanine.

WO 2017/ 182688 describes a saffron extract for the treatment of mood disorders.

Kaylynn Chiarello-Ebner: “Getting a Head Start - A report on the nutritional building blocks for memory and mood support”, Wholefoods Magazine, 2013-06-01 describes a saffron extract and a magnolia extract for use in the treatment of depression.

The problem underlying the present invention is to provide a composition for the treatment of mood and/or anxiety disorders containing active ingredients of natural origin, having an excellent tolerability, no side effects and an improved action compared to the compositions of the prior art.

Summary of the invention

The present invention solves the above-mentioned technical problem by providing a composition comprising 10-20 mg of saffron extract, 50-70 mg of magnolia extract and 180-220 mg of green tea extract for the use in the prevention and treatment of mood disorders and/or anxiety disorders.

Preferably, the composition further comprises L-theanine.

Preferably, the composition further comprises melatonin.

In another aspect, the present invention also describes a pharmaceutical or food supplement formulation suitable for oral administration, for the prevention and treatment of mood disorders and/or anxiety disorders, comprising the above-mentioned composition for the above-mentioned use and at least one pharmaceutically or nutritionally acceptable excipient.

Preferably, the above-mentioned formulation also comprises 180-220 mg of L-theanine and possibly also 0.5-3.0 mg, preferably 0.5- 1.5 mg of melatonin.

The above-mentioned quantities for the active ingredients of the formulation according to the invention correspond to the daily dose of each of them.

Preferably, the at least one pharmaceutically or nutritionally acceptable excipient is selected from the group comprising inert diluents, sweetening agents, flavors, anti-caking agents, lubricants, dyes, film-forming agents, microcrystalline cellulose, cellulose derivatives and thickening agents.

In a further aspect, the formulation according to the present invention is in the form of tablet containing the above-mentioned formulation.

Preferably, the tablet comprises an outer coating and an inner core comprising gastro-resistant granules containing said saffron extract and said magnolia extract.

Preferably, the gastro-resistant granules comprise polyvinylpyrrolidone and microcrystalline cellulose.

Preferably, the inner core further comprises a homogeneous mixture in the form of powder comprising said green tea extract.

Preferably, the homogeneous mixture in the form of powder further comprises said L-theanine.

Preferably, the homogeneous mixture in the form of powder further comprises further carboxymethylcellulose or salts thereof, microcrystalline cellulose and silicon dioxide.

Preferably, the outer coating comprises melatonin.

In an embodiment, the tablet has an inner core containing gastro- resistant granules, comprising saffron extract and magnolia extract, and a powder mixture comprising green tea and theanine extract, as well as an outer coating containing melatonin, and it is characterized in that it has an immediate release of melatonin, followed by a gradual release of green tea extract and theanine and a delayed release of saffron extract and magnolia extract.

The delayed release of saffron and magnolia extract occurs in the intestine.

The composition of the present invention advantageously comprises different naturally-occurring active ingredients which act in a complementary way and in synergy with each other.

In particular, the saffron extract rich in safranal and crocin exerts a serotoninergic action, thus directly contributing to the increasing of serotonin levels in the organism, thus considerably improving the patient’s mood.

Instead, the magnolia extract is provided with tranquilizing , calming properties, because it is able to considerably reduce states of anxiety, sleeping alterations and mood instability.

The green tea leaves extract, rich in catechins, shows an important antioxidant activity and reduces the oxidative stress, to specifically reduce anxiety conditions and sleep disorders.

According to an embodiment, this composition advantageously also comprises L-theanine amino acid, which directly affects the noradrenalin, serotonin and dopamine levels in the organism. In particular, L- theanine’s main action is on the anxiety reduction, acting as an antagonist of the excitatory neurotransmitters (for example glutamic acid) and thus favouring a feeling of relaxation of the patient.

According to a further embodiment, the composition of the invention further comprises melatonin as naturally-occurring active ingredient. The main effect of melatonin is the circadian rhythm regulation, thus favouring falling asleep and sleep continuity. Furthermore, since it has an antioxidant effect, it acts in general by lowering the oxidative stress and consequently it favours an alleviation of the anxious state disorders.

Advantageously, the composition according to the invention favours the recovery of psycho-physical well-being in patients who suffer from mood disorders, in particular from mood lowering, due to the synergistic and complementary action of the active ingredients contained therein.

Furthermore, it has been noticed that the intake of such a composition in the form of food supplement favours a reduction of the state of anxiety, thus reducing both symptoms related to the emotional sphere, such as fear, apprehension and concern, and physical symptoms, such as palpitations, chest pains, shortness of breath and/or nausea. A further advantage of the present invention is that such composition may also have a beneficial effect on sleep disorders.

Furthermore, the composition described herein does not display certain side effects typical of the above-mentioned synthetic drugs administered to patients who suffer from mood disorders, such as depression.

The formulations for the oral administration comprise for example capsules, tablets, powders, granules and gels.

In such solid dosage forms, the active compounds can be mixed with different excipients, such as inert diluents, such as for example saccharose, lactose, starch or maltodextrin, sweetening agents, such as sorbitol, sucralose, steviol glycosides, flavors, anti-caking agents, such as silicon dioxide, lubricants, such as for example magnesium salts of saturated fatty acids, dyes, film-forming agents, microcrystalline cellulose, cellulose derivates, such as carboxymethylcellulose, ethyl cellulose, hydroxypropyl methylcellulose and thickening agents.

The pharmaceutical preparations according the present invention can be produced by using conventional pharmaceutical techniques, as described in the different pharmacopoeias or in handbooks of the field, such as for example “Remington’s Pharmaceutical Sciences Handbook”, Mack Publishing, New York, 18th Ed., 1990.

The average daily dosage of the composition according to the present invention depends on many factors, such as for example the patient’s conditions (age, weight, sex): the total dose of the active ingredients may vary in general from 240 mg to 533 mg per day, possibly divided into more than one administration.

According to a preferred embodiment, such a composition may be formulated in tablets that allow a multiphasic release of the active ingredients therein contained.

Advantageously, such a formulation allows a release of the active ingredients at scheduled times to obtain a gradual beneficial and long- lasting effect.

A further advantage is represented by the fact that the present formulation can favour a considerable reduction of the oxidative stress (at a systemic level) which is closely related to anxiety and/or depressive states, which are often characterized by insomnia.

Brief description of the figures

Figure 1 shows the release profiles over time of tablets according to the present invention immersed in a HC1 solution (pH 1.1) for 120 minutes and later immersed in a phosphate buffer 0.2 M (pH 6.8) solution for 3 hours.

Figure 2 shows a bar graph of the scores according to the test DASS-21 obtained from each subject of group A before the treatment and after the treatment. The score difference (delta) between the score obtained before the treatment and that obtained after the treatment is further shown.

Figure 3 shows a bar graph of the score averages according to the test D ASS-21 of group A obtained before the treatment and after the treatment.

Figures 4A and 4B show two graphs which report the percentage distribution of the subjects of group A according to the test DASS-21 , in the score range 0-21 (no depressive traits) and in the score range 22-42 (moderate depression), respectively, before and after the treatment.

Figure 5 shows a bar graph of the scores according to the test DASS-21 obtained from each subject of group B before the treatment and after the treatment. The score difference (delta) between the score obtained before the treatment and that obtained after the treatment is further shown.

Figure 6 shows a bar graph of the score averages according to the test D ASS-21 of group B obtained before the treatment and after the treatment.

Figures 7 A and 7B show two graphs which report the percentage distribution of the subjects of group B according to the test DASS-21 , in the score range 0-21 (no depressive traits) and in the score range 22-42 (moderate depression) before and after the treatment, respectively.

Figure 8 shows a bar graph of the scores according to the test STAI-Y obtained from each subject of group A before the treatment and after the treatment. The score difference (delta) between the score obtained before the treatment and that obtained after the treatment is further shown.

Figure 9 shows a bar graph of the averages of the scores according to the test STAI-Y of group A obtained before the treatment and after the treatment.

Figures 10A and 10B show two graphs which report the percentage distribution of the subjects of group A according to the test STAI-Y, in the score range 20-40 (no significant anxiety problem) and in the score range 41-60 (physiological and moderate anxiety), before and after the treatment, respectively.

Figure 1 1 shows a bar graph of the scores according to the test STAI-Y obtained from each subject of group B before the treatment and after the treatment. The score difference (delta) between the score obtained before the treatment and that obtained after the treatment is further shown.

Figure 12 shows a bar graph of the averages of the scores according to the test STAI-Y of group B obtained before the treatment and after the treatment.

Figure 13A and 13B show two graphs which report the percentage distribution of the subjects of group B according to the test STAI-Y, in the score range 20-40 (no significant anxiety problem) and in the score range 41-60 (physiological and moderate anxiety) and in the score range 61-80 (anxiety predominating in everyday life) before and after the treatment, respectively.

Detailed description

The present invention describes a composition for the prevention and/or treatment of mood disorders and/or anxiety disorders.

The expression“mood disorders”, as used herein, means in particular the depressive symptoms and the pathologies therewith associated, such as for example the lowering of mood, sleep disorders, persistent sensation of sadness, difficulty in concentrating, increase or reduction of appetite. Physical symptoms determined by the depressive state, among which for example tremor of the limbs, dry mouth, tachycardia, increased sweating and difficulty in swallowing, are further comprised.

The expression“anxiety disorders” or“states of anxiety”, as used herein, means in particular disorders related to the state anxiety and trait anxiety. The state anxiety is conceived as a particular experience, a feeling of insecurity, impotence in front of a perceived harm, which can lead to concern or escaping and avoiding. The trait anxiety consists of the tendency to perceive stressful situations as dangerous and threatening and to respond to the various situations with different intensity.

This composition exclusively contains active ingredients of natural origin, namely saffron extract, magnolia extract and green tea extract and can optionally contain also L-theanine and melatonin.

The expression“saffron extract”, as used herein, means a Crocus sativus (stigmas) dry extract titrated in crocin and safranal (expressed as sum, at 3.5%).

The expression“magnolia extract”, as used herein, means a Magnolia officinalis (bark) dry extract titrated in honokiol (> 2%).

The expression“green tea extract”, as used herein, means a Camellia sinensis (green leaves) dry extract titrated in total catechins (> 60%) and epigallocatechin gallate (> 40%) and decaffeinated.

In particular, the composition, as its active ingredients are all of natural origin, shows excellent tolerability and is substantially free of side effects, in particular of those side effects that are typical of the synthetic drugs that are usually administered to contrast mood disorders.

Furthermore, such composition comprises different active ingredients which have different effects and act in synergy with each other; for this reason, the composition of the invention favours a total recovery of the psycho-physical wellbeing of patients who suffer from sleep disorders, thus improving the mood and, at the same time, alleviating anxiety disorders and favouring falling asleep and sleep continuity.

The composition according to the present invention comprises: saffron extract, magnolia extract and green tea extract.

According to a preferred embodiment of the present invention, the composition is formulated in tablets and is taken as food supplement with controlled release.

Saffron extract

The saffron extract obtained from the stigmas of such plant contains different compounds, among which safranal and crocins.

In particular, crocins are the most abundant components in the saffron plant and can exist in different isoforms, the most abundant of which is alfa-crocin.

These compounds are glycosylated carotenoids and have a quite particular structure; they are actually constituted by 20 carbon atoms with 2 carboxylic groups on the first carbon 1 (Cl) and on the sixteenth carbon (Cl 6).

The presence of crocins in the saffron extract seems to be essential for the action of this extract in improving the mood in patients suffering from mood disorders.

In particular, the saffron extract acts by means of the so-called “serotonergic action”, namely an inhibition mechanism of the serotonin reuptake at presynaptic nerve receptors, thus increasing the availability of this molecule in the spaces designated for the nervous transmission (synapses).

The consequent local increase in serotonin has as main effect the improvement of the patient’s mood.

Furthermore, since the saffron extract is rich in carotenoids, it also shows an antioxidant and neuroprotective action, thus reducing the levels of the reactive oxygen species (ROS) in the organism.

Saffron pharmacokinetic

Crocin is not directly absorbed but is hydrolysed in crocetin, which can be absorbed at the intestine .

Crocetin has an interesting pharmacokinetics which significantly differs from that of the other known carotenoids naturally present in foods or contained in nutraceutical products, such as for example beta carotene, lutein and lycopene.

It has been observed that crocetin is rapidly absorbed and its presence can be noticed in plasma around one hour after the administration reaching a peak at around 4 hours. Such a compound is characterized by a quite long half-life of 6-7 hours since, after its fast absorption, a phase of around 4 hours of slower and more gradual absorption follows.

Furthermore, the interaction with serum albumin is quite weak and this facilitates a good distribution towards the peripheral tissues of the organism.

Such a hypothesis of crocetin absorption differs from that of the other carotenoids which, being part of the chylomicrons, cross the intestinal cells by passive diffusion, enter the lymphatic circulation and later the blood circulation. Crocetin is more hydrophilic and smaller than other carotenoids, therefore it is directly absorbed by the portal vein and therefrom it is distributed into the organism passing through the liver.

Its main metabolite derives from the conjugation with glucuronic acid.

Magnolia extract

Magnolia extract contains magnolol and honokiol, two phenolic compounds which act by the modulation of physiological mechanisms triggered by stress.

Magnolia extract contributes to the reduction of sleep disorders, thus improving the sleep quality and favouring a perceivable decrease of the daily sleepiness.

This extract as well exerts a strong GABAergic action which can be compared to that exerted by the benzodiazepines (the so-called hypno- inducing molecules), but which does not have the side effects thereof.

Magnolol and honokiol modulate the GABA-A receptors of the cerebral limbic system, centre of emotions and feelings; this explains the calming and muscle-relaxant activity and the rebalancing activity of the sleep alterations. These effects determine a perceivable reduction of the mood disorder, irritability, anxiety symptoms and also of sleep disorders.

Magnolia extract pharmacokinetics

After the oral administration of magnolia dry extract, it has been observed that the main metabolite is the conjugate of hydroxy-magnolol. Furthermore, it has been observed that the magnolia extract naturally contains around 94% of magnolol and 1.5% of honokiol. Consequently, it is believed that the calming effect attributed to this extract is due to the presence of magnolol.

Green tea extract The green tea extract contains epigallocatechin gallate (EGCG) and catechins, which are antioxidant molecules that contribute to reduce the oxidative stress, a target specifically studied to reduce anxiety conditions, depression and sleep disorders.

In particular, the possible mechanism of action of the EGCG at a cerebral level occurs on the receptor GABA-A through the carbonyl portion of the molecule, which is generally common to the flavonoids.

This binding has a sedative effect in conditions of acute stress, reducing the vigilance time (vigil) and exerting a sedative and hypnotic action.

Green tea pharmacokinetics

The pharmacokinetic profile of polyphenols after oral ingestion is very interesting because the original compounds do not appear in the circulation but phase-II metabolites do, and their presence in circulation, after the consumption with foods, rarely exceed the nanomolar concentrations .

Significant quantities of both the original compounds and the metabolites reach the colon, where they are further decomposed by the microflora, mainly yielding small phenolic acids and aromatic catabolites which are absorbed in the systemic circulation.

With regard to the oral bioavailability, flavonoids are typically ingested with the diet in the glycosylated form and their absorption is associated with a cut and release of the aglycone as a result of the action of enzymes located on the apical wall of the small intestine.

The cleavage enzymes have affinities to the specific chemical structures and the bonds between the sugar unit and the aglyconic part. The latter could be absorbed by passive diffusion through the epithelial cells as a result of the increase of its lipophilicity.

Another hydrolytic stage could be mediated by cytosolic beta-glycosidase inside the epithelial cells. Therefore, in this case, the polar glycoside would be transferred into the cell through the involvement of the transfer of the sodium-dependent glucose transporter type 1 (SGLT1).

Before the passage in the bloodstream, the aglyconic component undergoes a phase II metabolism, , forming sulphate, glucuronate and/or methylated conjugates through the correspondent enzymes involved in these reactions (SULTs, UGT, COMTs).

Furthermore, also outflow phenomena occur whereby some metabolites go back to the small intestine lumen through specific reverse transporters, among which P glycoprotein.

L-theanine

L-theanine amino acid affects the levels of noradrenaline, serotonin and dopamine. In particular, it acts as antagonist of the glutamic acid, an excitatory neurotransmitter, thus reducing the state of anxiety.

L-theanine is rapidly absorbed at the intestinal level and distributed in the tissues and, in particular, it very easily trespasses the human blood- brain barrier through the transport system of the neutral amino acids.

L-theanine pharmacokinetics

L-theanine amino acid reaches the maximum plasmatic concentration between 30 minutes and 2 hours after the administration.

The main catabolite is ethylamine, which is excreted and observed at the renal level.

Such amino acid produces different variations in the cerebral biochemistry and in particular in the synthesis or inhibition of some neuro transmitters .

L-theanine in fact increases the levels of GABA, responsible for the feeling of relaxation and the so-called “wellbeing effect”, and it exerts a significant effect on the release of dopamine and serotonin, which are closely related to the mnemonic and learning abilities.

The amino acid acts by reducing the glutamate release from the presynaptic neurons, which acts as powerful excitatory neuro transmitter and increases the cerebral alpha waves which induce relaxation.

Furthermore, it reduces the concentration of alfa-amylases in the saliva and counters the effects of caffeine; this last mechanism favours an improvement in sleep quality.

Melatonin

Melatonin is a hormone which is mainly released by the hypophyseal or pineal gland located at the base of the brain.

Serotonin is one of the main precursors of melatonin, whereby in a patient suffering from mood disorders, in which the concentration of serotonin is quite low, the concentration of melatonin is also reduced, therefore causing sleep disorders or worsening them.

The melatonin supplements are thus administered to these patients in order to compensate for the deficiency of melatonin naturally produced by the organism and to regulate the circadian rhythm.

Furthermore, melatonin also carries out an antioxidant action, protecting the cells from the action of free radicals, whose concentration increases in stress and insomnia conditions.

Melatonin pharmacokinetics

Melatonin has an oral bioavailability characterized by a high interindividual variability: 3%-76%.

It has an absolute bioavailability of 15% for dosages of 2 and 4 mg.

The oral bioavailability parameter is significantly affected by the extent of the intestinal absorption and the extent of the metabolic elimination processes. In fact, the absorbed melatonin undergoes a significant hepatic first pass metabolism of: phase I: hydroxylation from cytochromes; phase II: conjugation with sulphate or hyaluronic acid.

The absorption can even diminish by 50% in the elderly subject.

Melatonin is distributed in the peripherical tissues due to its lipophilicity and is eliminated in the form of metabolites through urine, saliva and milk.

The half-life time of melatonin is around 30-60 minutes, whereby it does not undergo accumulation phenomena.

The formulation of the composition

According to the present invention, the above-mentioned composition is preferably formulated in tablet and administrated as food supplement.

This tablet is structured so as to allow a release programmed over time of the single active ingredients therein contained.

In particular, this tablet has an outer coating and an inner part consisting of a core which comprises in turn a gastro-resistant granulate inside it.

According to a preferred embodiment, the outer coating consists of a film comprising melatonin as active ingredient; the core comprises the green tea extract and L-theanine and the granulate comprises the saffron extract and the magnolia extract.

Such tablet structure allows to obtain a release in subsequent phases of the different active ingredients, thus allowing obtaining a gradual and long-lasting beneficial effect. In particular, melatonin is the first active ingredient released in the organism, since it is comprised in the most external part of the tablet.

Hereafter, the release of L-theanine and the green tea extract contained in the core occurs, and finally the release of the active ingredients contained in the gastro-resistant granulate, namely the saffron extract and the magnolia extract (Example 1), occurs.

The tablet preparation method involves several steps.

The first step involves the preparation of a gastro-resistant granulate, by mixing the saffron extract and the magnolia extract with polyvinylpyrrolidone and microcrystalline cellulose, and adding then a granulation liquid and the shellac ammonium salt.

Then the preparation of the tablet core is performed.

The gastro-resistant granulate obtained as above described is mixed with the following ingredients, which have been previously singularly sifted: green tea extract, L-theanine, cross-linked sodium carboxymethylcellulose, microcrystalline cellulose and silicon dioxide; then, the flowability and density of the mixture is verified.

After the core has been prepared, it is pressed, thus obtaining non-coated tablets with weight of around 1 g each one.

The last preparation step is the coating step.

To prepare a coating film, water and glycerine are mixed to which white Sepifilm® LP 770 (made of hydroxypropyl methylcellulose, microcrystalline cellulose, fatty acids and titanium dioxide), a dye, glycerine and melatonin are added. After mixing for at least 40 min, the possible formation of lumps is controlled, and then a filtration is performed until a homogeneous mixture is obtained.

Afterwards, the coating of the tablets is performed by pan-coating under constant stirring. The average weight of the coated tablets is around 1.1 g each. EXAMPLE 1

Tablet composition according to the present invention

The quantity of used purified water is the one necessary and sufficient for the granulation.

EXAMPLE 2 - Controlled release test

The tablets obtained according to the method of the present invention have been tested in vitro for determining whether they had been structured so as to allow a controlled release over time of the active ingredients contained therein.

The release profile has been determined through samplings at 1 , 30, 60, 90 and 120 minutes of tablets immersed in a HC1 solution (pH 1.1), in order to mimic the gastric environment and, subsequently, every 30 min in a 0.2M phosphate buffer (pH = 6.8) in order to mimic the intestinal environment for 3 hours.

The results are illustrated in Figure 1 , which shows the dissolution profiles obtained as an average of three independent experiments and reported as release percentage with respect to the total titre.

It is evident that the tablet so structured allows a programmed and gradual release over time of the different active ingredients.

The first released active ingredient is melatonin, which is comprised in the outer coating film, followed by L-theanine (comprised in the core) and by the saffron extract (comprised in the gastro-resistant granulate).

EXAMPLE 3 - THE CLINICAL TRIAL ON THE DEPRESSIVE SYMPTOMS

A pilot clinical trial has been carried out involving 24 healthy volunteers to evaluate both the previous presence of mood and anxiety symptoms and the possible improvement thereof due to the intake of a tablet containing saffron extract, magnolia extract, green tea extract and excipients as described in the present invention.

The effects obtained with the above-mentioned tablet have been compared to the effects obtained with a tablet formulated according to patent application ES 2573542, which describes a composition only containing saffron extract (28 mg) as active ingredient for the treatment of the mood disorders related to depression.

The aim of the present trial was that of demonstrating that saffron at a lower dosage, and associated with other naturally occurring ingredients, such as the magnolia extract and the green tea extract, which are per se not known to influence the mood, had a synergistic effect on the improvement of the depressive symptoms and possibly also of the anxiety symptoms, compared to the effect of the saffron extract used alone.

The subjects

24 subjects attended the trial: : 10 men and 14 women of age between 30 and 70 years (average age 46 years).

The 24 chosen subjects did not suffer from serious diseases and were not taking drugs that could potentially interact with the treatments. Four subjects have been excluded because they did not complete the trial.

The grouys

Two groups were randomly formed as follows:

1) Group A: 9 subjects (5 males and 4 females), who took a tablet according to the invention described in Table 1.

2) Group B: 1 1 subjects (3 males and 8 females), who took a tablet according to patent application ES 2573542 described in Table 1.

For both groups, the posology was of 1 tablet a day before bedtime, for 24 consecutive days .

The tablets Table 1 : Tablet composition

The tablet obtained according to the present invention contains three naturally-occurring active ingredients: green tea extract, saffron extract and magnolia extract. This tablet was administered to group of volunteers “A”.

The tablet obtained according to patent application ES 2573542 only contains saffron extract as naturally-occurring active ingredient. This tablet was administered to group of volunteers“B”.

The questionnaire for depression sumytoms In order to gather useful informations before and after the trial and to quantitatively compare them with each other, a literature-based questionnaire has been worked out. .

The work by Bottesi, Ghisi et al., 2015, The Italian version of the Depression Anxiety Stress Scales-21 : Factor structure and psychometric properties on community and clinical samples has been considered as the reference for assessing the possible disorders related to the mood disorders.

The Depression Anxiety Stress Scales-21 (DASS-21) is a shorter version of another self-assessment questionnaire that was originally developed to make a differential diagnosis between depressive and anxiety symptoms. The cited work demonstrated the validity and usefulness of the Italian DASS-21 , both for clinical communities and clinical individuals. For this reason, it has also been chosen in the trial described herein.

The“DASS-21” questionnaire is composed as shown in Table 2:

Table 2: Questions, answers and score of the“DASS-21” questionnaire

The score was expressed as the sum of the 21 questions for each participant. The interpretation of the results follows this scheme:

The results of grouy A

The 9 subjects of group A started from an average score of the DASS-21 questionnaire equal to 17.7 corresponding to“no depression However, 8 subjects out of 9 had an improvement of the initial condition related to their perception of mood (Figure 2).

Subject E, who did not report a numeric improvement, seems to confirm the score obtained before the treatment, which could be explained as a variability which is subjective and not indicative of an exacerbation. Considering the scores of each subject individually, subjects B, D and I had an initial score falling within the score range“22-42”, corresponding to“lowering of mood and moderate depression”. Their score after the treatment was considerably reduced, namely:

B: from 27 to 3 (delta -24); - D: from 25 to 1 (delta -24);

I: from 27 to 15 (delta - 15).

As shown in Figure 3, the average score of group A according to the results obtained from the DASS-21 test decreased by 10.22 points (- 57.84%) after the treatment, indicating an objective perception of the mood improvement by the participants.

Figure 4B shows that all participants fall within the score range“0-21” after the treatment, whereas around 33% of the participants was in the score“22-42” (Figure 4A) before the treatment.

No participant in the trial reported mild, moderate or serious side-effects related to the use of the product.

The 1 1 subjects of group B started from an average score of the DASS-21 questionnaire equal to 15.27, corresponding to“no depression”.

However, 9 subjects out of 1 1 had an improvement of the initial condition related to their mood perception (Figure 5).

Subject Q confirmed his initial score.

The average score of DASS-21 after the treatment decreased by 4.91 points (-32.15%) indicating a modest improvement of the participants’ mood (Figure 6).

Figures 7 A and 7B show that the number of subjects with score“0-21” (around 91%) considerably increased after the treatment with the tablet of the present invention, compared to that observed before the treatment (around 64%).

No participant in the trial reported mild, moderate or serious side effects related to the use of the product.

The results obtained show that a considerable reduction of the depressive symptoms was achieved. In fact, a reduction of the average value of DASS-21 by 57.84% in group A has been observed; the average value has therefore been halved compared to the beginning and is considerably greater than the reduction by 32.15% in group B.

These results show that the composition of the present invention, containing saffron extract (in a quantity equal to 15 mg), magnolia extract and green tea extract has a synergic action on the depression symptoms compared to the known composition only containing saffron (in a quantity equal to 28 mg) as active ingredient.

EXAMPLE 4 - THE CLINICAL TRIAL ON ANXIETY DISORDERS

The subjects, groups and tablets are the same as those described in Example 3.

The questionnaire for anxietu disorders

As for anxiety disorders, the self-assessment questionnaire commonly known as“STAY TRAIT ANXIETY INVENTORY” (STAI-Y) has been used as reference.

The STAI-Y is a tool easy to apply and interpret for detecting and measuring anxiety, both for psycho-diagnostic purposes and for verifying efficacy and benefits of the psychotherapy or another treatment.

The questionnaire is formed by 40“items”, which the subject has to answer to in terms of intensity (from“hardly never” to“almost always”) . The items are grouped into two scales focused on how the subjects usually feel or on what they instead feel in particular moments.

The Applicant simplified this basic structure by reducing the 40“items” to 20, but preserving the structure of the two scales which are the following:

• State anxiety, wherein anxiety is understood as a particular experience, a feeling of insecurity, impotence in respect of a perceived damage which can lead to concern or to escape and avoidance. The state anxiety is measured through questions referring to how the subject feels at the moment of the questionnaire submission.

• Trait anxiety, which consists in the tendency to perceive stressful situations as dangerous and threatening and to respond to the various situations with different intensity. Therefore, questions are asked which inspect how the subject usually feels. This tool is useful to carry out a generic measure of stress, not establishing direct links to work situations.

The two scales are shown in detail hereinafter with the related questions, answers and score to be attributed thereto (Tables 3 and 4). Table 3: Evaluation scale for the state anxiety.

Table 4: Evaluation scale for trait anxiety.

The score was expressed as the sum of the two scales for each participant. The interpretation of the results follows this scheme:

The results of grouy A

GROUP A:

The 9 subjects of group A started from an average score of the STAI-Y questionnaire (adapted according to the above simplification of the Applicant) equal to 44.33 corresponding to“a physiological, moderate, manageable anxiety”.

8 subjects out of 9 (89%) had an improvement of the initial condition.

Among them, 4 subjects (A, B, G, H) fell within the score range“20-40” corresponding to“no significant anxiety problem” with a great decrease of their initial score (delta A (- 18); delta B (- 13); delta G (- 16); delta H (- 19).

Other 2 subjects (F, N) remained in the score range “20-40” though improving the initial value, and 2 subjects (D, I) remained in the range “41-60”. Only one subject (indicated with letter E) did not improve the initial condition; he / she started from a score of 33 and reported a score of 36 at the end of the treatment. This change should not be considered as worsening but rather as a subjective variable (Figure 8).

The average score of STAI-Y after the treatment decreased by 9.44 points (-21.29%), passing from the intermediate score range to the desired score range (Figure 9).

As shown in Figures 10A and 10B, only 33% of the patients fell within the score“20-40” before the treatment; instead, this percentage reaches 78% of the patients after the treatment, therefore indicating that the tablet containing the composition of the present invention also has a beneficial effect in the treatment of anxiety disorders.

The 1 1 subjects of group B started from an average score of the STAI-Y questionnaire equal to 47.45 corresponding to“a physiological, moderate, manageable anxiety”.

8 subjects out of 1 1 had an improvement of the initial condition but not always such as to fall within the score range corresponding to “no significant anxiety problem”.

In fact, 6 subjects (P, T, U, Z, X, Y), which showed an improvement, still remain in the score range of 41-60 corresponding to “physiological, moderate, manageable anxiety”. The other two subjects (Q and W) fall within the desired range (Figure 1 1).

3 subjects (R, S, V) do not improve the initial value.

The data related to subject R is interesting, since an increase of 15 difference points is observed, which can be indicative of a worsening condition. As for subject S, in this case like in group A, the increase is not to be considered as worsening, but it rather reflects the subjective variability. Subject V substantially confirms the initial condition without any improvements.

The average score of STAI after the treatment decreased by 3.64 points (-7.7%) (Figure 12).

None of the participants in the trail reported mild, moderate or serious side effects related to the use of the product.

Figure 13 A shows the distribution of the subjects of group B according to the STAI score before the treatment: 18% of patients are in the score 20-40. Instead, Figure 13B shows the distribution of the same subjects after the treatment with a tablet comprising the composition described in known document ES2573542.

In particular, it is noticed that around 9% of the subjects showed a worsening of the anxiety disorders and that, generally, a little increase in the percentage of the subjects in the score 20-40 takes place.

Moreover, considering the values of each participant, the results indicate an overall improvement of group A compared to group B. In fact, in group A at the beginning only 33% of the subjects fall within the range“20-40”, after the treatment 78% of the subjects fall within this range.

These results indicate that the composition of the present invention containing saffron extract (in a quantity equal to 15 mg), magnolia extract and green tea extract has a synergic action on the anxiety disorders compared to the known composition only containing saffron (in a quantity equal to 28 mg) as an active ingredient.

Such a composition is therefore effective not only for the treatment of depression disorders but also for the treatment of anxiety disorders.

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