"Composition for use in the prevention and/or treatment of symptoms related to tiredness and fatigue"

In the name of: Neilos Sri of Italian nationality

Residing in: via Bagnulo 95 - 80063 - Piano di Sorrento

(NA)

Designated inventor: Umberto Di Maio

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DESCRIPTION

The present invention relates to a composition of substances effective in the prevention and/or treatment of symptoms related to mental or physical tiredness and fatigue .

DESCRIPTION PHYSICAL AND MENTAL FATIGUE

Under the terms of tiredness and fatigue reference is made to different conditions due to several causes each individual can come across during his/her own life.

In particular, generally "fatigue" commonly is divided into mental fatigue and physical fatigue.

The mental fatigue is a very common psychobiological state in the daily life. Such physio-pathological condition is characterized by a general sensation of tiredness, fatigue and lack of energies and it has consequences affecting, almost immediately, the correct operation of the primary cognitive functions. The appearance of this general fatigue sensation is accompanied by difficulty in concentration and in keeping the normal attention levels and, in some cases, it can jeopardize the normal daily activities, the working performances as well as the physical performances. Many studies have underlined that the attention, which is one of the fundamental features in the development of a dynamic behaviour and the main tool for selecting information, is negatively influenced by the mental fatigue. The factors at play underlying this sensation of fatigue and tiredness can be many: prolonged periods of a high cognitive requirement; the presence of a pathology, such as anaemia and thyroid disorders; deficiency of some essential micronutrients; excessive sedentary or excessive physical activity and above all oxidative stress. Such condition is characterized by the rupture of redox equilibria inside the cells and by the accumulation of pro-oxidant reactive radical species which, although they are involved in important physiological mechanisms, such as defence against bacteria, at high levels can damage the cellular components. The free radicals can attack the polyunsaturated fatty acids of the membranes (by causing a chain of peroxidation reactions), proteins and enzymes (by damaging the functionality thereof) , the nucleic acids (by causing the filament rupture and alteration of nitrogenous bases) and, above all, the mitochondrial DNA which is not protected by the histones. Then, they can alter physiological cellular mechanisms even at central level. The connection between oxidative stress and damage of the cognitive functions was demonstrated. Some studies demonstrated the presence of high plasmatic levels of the main markers of the oxidative stress in subjects with chronic fatigue syndrome and in subjects with Burnout syndrome, and it is known that high levels of free radicals can be found in the neuro- degenerative pathologies, such as Parkinson and Alzheimer. The mental fatigue might affect even physical performances and, then, be associated to a sensation of physical tiredness. The physical fatigue can be defined, independently from the mental fatigue, as fatigue and tiredness deriving from a performed effort and can be re connected to lack of sleep, lack of energies and to a temporary malfunction of the muscles, in turn caused by a tissue damage which can be ascribed to more or less intense physical activity. Although the beneficial effects of physical activity are known, there is a considerable number of tests indicating that the physical exercise, in particular of high intensity and duration, leads to a muscle oxidative damage, even in trained individuals. The idea that exercise increases the production of free radicals is confirmed by studies which have found an increase in indicators of oxidative damage in muscle and cardiac tissue and a decrease in GSH/GSSG ratio, data indicating the increase in oxidized glutathione and, consequently, the increase in the pro-oxidizing reactive species. The oxidative stress, deriving from an excessive formation of radical species at muscle level, is associated to fatigue and muscle lesions, both during contraction and during the recovery phase. It was observed that when the levels of free radicals are too high, the muscle force reduces and fatigue increases. One of the mostly involved factors in this mechanism is the mitochondrial alteration.

The damages to mitochondrial DNA cause alterations in the complexes of electron transport chain and, consequently, difficulty in ATP synthesis. In order to sustain the energy levels, the cellular metabolism moves towards the anaerobic route rather than the aerobic one. As a consequence, there will be an increase in the lactic acid which is one of the main factors responsible for the muscle fatigue.

Other conditions which generally are linked to physical and/or mental tiredness are: asthenia, loss of appetite, malnutrition, convalescence from drug therapies, senile dementia, Alzheimer disease, neuronal aging, mild cognitive impairment, attention deficit, work-related stress, psycho physical stress.

The composition, the present invention relates to, is inserted in this context, which composition comprises an association of at least an extract of Schisandra chinensis, at least an extract of Morinda citrifolia (commonly called Noni), inositol and at least an extract of a plant belonging to the genus Cola. Such composition is particularly effective in the prevention and/or treatment of symptoms related to mental or physical tiredness and fatigue .

SCHISANDRA CHINENSIS

Schisandra chinensis is a climbing shrub belonging to the family of Schisandraceae, which can reach sizes ranging from 0.5 to 25 m of length. The plants belonging to this species are little resistant both to extremely humid and dry environments, therefore they prefer tropical or subtropical environments, with light and humidity moderate conditions and a humus-rich soil. Schisandra chinensis is endemic in North-East of China and in Korea wherein it has always been used for its beneficial properties; in particular in the Chinese popular medicine it is used as stimulant in tiredness and sexual impotence and as tonic in indolence and physical tiredness. Such shrub grows spontaneously even in Eastern Russia wherein it has been consumed for decades by native populations to improve the night vision and as tonic to reduce hunger, thirst and tiredness. The phytochemical analyses, performed to establish the nature of the active principles responsible for the tonic actions of Schisandra, revealed the presence of traces of ethereal oils, vitamin C, tannins and a high content of lipidic components. In particular in berries one found the presence of a high content of citric, malic and tartaric acid, several microelements and lignans, a group of polyphenols in which the main components are Schisandrin, g-Schisandrin and Gomisine. Such polyphenols are the main responsible for the tonic effect and for the improvement of the physical and mental performances in healthy subjects. In fact, Schisandra results to be an interesting stimulating of SNC and its tonic action can be used in case of exhaustion and stress situations to reduce the physical and mental tiredness and to improve the working capabilities. Moreover, Schisandra increases force and physical resistance.

An in-vivo study showed the effect of lignans of Schisandra in mice treated with 0.5 mg/kg of Scopolamine (muscarinic antagonist) . The treatment caused in mice cognitive and memory impairment, detected with eight-arm maze test; Schisandrin, at the dose of 1 and 10 mg/kg, annulled Scopolamine effect by reducing the residence time in the maze and by decreasing the number of errors. Moreover, Schisandrin at the same doses, increased the tremors induced by oxotremorine (muscarinic agonist Mi) . Analogous results are shown by an in-vivo study on mice with cognitive impairment induced by Scopolamine 1 mg/kg. The study evaluated the effect of cognitive improvement of Gomisine A by means of Y-maze and Morris water maze test. A treatment with 5 mg/kg of Gomisine A annulled the scopolamine effect. These results suggest that Schisandra effect on SNC is mediated by a strengthening of the cholinergic system.

A clinical study on telegraph young operators aged between 21 and 24 tested parallelly the extract of Schisandra and Schisandrin. Two sets of experiments were prepared:

N1 : (20) extract of Schisandra 30 mL (10% in 70% ethanol ) ;

- N2 : (23) Schisandrin 5, 10 and 20 mg.

The subjects were asked to transmit the Morse code for five minutes at the maximum speed and the error frequency was evaluated. In the treated group, after one single administration, the error frequency is in a range between 84 and 103% with respect to the control group, treated with placebo (130%) . The results show that Schisandra improves the mental performances and the working capabilities by reducing the errors related to tiredness.

A clinical study on subjects subjected to intense physical effort demonstrated that the administration of Schisandra (1 g per day) increases the physical capabilities. By means of Dubua test an increase in the working capabilities from 27.5 kg/m in the control group to 77 kg/m in the treated group was detected.

MORINDA CITRIFOLIA

Morinda citrifolia, also known as Noni, is an evergreen plant having small sizes belonging to the family of Rubiaceae. It can reach a height of 3 - 6 m as far as a maximum of 10 m. The ripe fruits are yellow and can reach sizes of 10 - 12 cm. The plant has a wide adaptability and it is widespread in all countries of the tropical area. Morinda is widely present in Indonesia and Polynesia and here it has been consumed by the local population for over 200 years for its several beneficial properties. The traditional medicine attributes Noni beneficial effects in many disorders such as osteoarthritis, rheumatisms, inflammation, diabetes, haemorrhoids, menstrual disturbance, infections, pain reduction, increase in the energy levels and well-being improvement. From the nutritional point of view, it results to be interesting for the content of minerals, amino-acids and vitamins. Several studies about Noni composition detected the presence of reducing sugars, phenols, tannins, flavonoids, saponins, glycosides, terpenoids, and alkaloids. Noni has a very high antioxidant power and it results to be useful in the states of physical and mental tiredness wherein the oxidative stress plays a predominant role. On this matter a study performed in double blind on 285 adult smokers, consuming more than 20 cigarettes per day, shows the powerful antioxidant action of Noni . The subjects were divided into three groups :

- Placebo group: grape and blackberry juice;

Group A: 29.5 mL of Noni juice;

Group B: 118 mL of Noni juice.

After thirty days of treatment the levels of SARS (superoxide anionic radical) and LOOH (hydroperoxide lipid) were evaluated. In the placebo group a significant decrease in the considered marker was not detected. In group A the levels of SAR and LOOH decrease by 26.9% and 24.5%, respectively. In group B the decrease in SAR and LOOH reaches 30.8% and 27.3%, respectively.

Additional studies performed on this species confirmed its positive effect on the well-being improvement, on the fatigue decrease and on the resistance increase.

In an in-vivo study on mice a comparison between the extract of Morinda citrifolia, standardized in Scopoletin and Epicatechin, and the extract of Morinda elliptica, standardized in Scopoletin only, was made. The mice were treated for six weeks with 200 and 400 mg/kg of each extract. The results showed that the extract of Morinda citrifolia is better than elliptica since it quintuples the resistance time, increases the levels of muscle glycogen, increases the glucose metabolism and lipid metabolism and delays the formation of lactic acid.

A series of clinical studies demonstrated the tonic effect of Morinda linked to its powerful antioxidant action. A study carried out on 40 trained athletes showed an increase of 21% in the tiredness average time and a decrease of 25% in the lipidic oxidation in the group treated with 100 mL of Noni juice twice a day, with respect to the group treated with placebo. From a study on 46 university athletes it appeared that the treatment with 100 mL of Noni juice twice a day causes a decrease in the concentration of Creatin Chinasi and in the tissue damage induced by the physical exercise, with consequent tiredness increase. In another study carried out on patients with advanced cancer, capsules containing 500 mg of dehydrated Noni were administered, starting from an initial dose of 2 g per day until reaching 8 g per day. The treatment led to a physical state improvement and to a better fatigue control by showing the powerful tonic effect of Noni.

INOSITOL

Inositol, or Myo-Inositol, is a six-carbon-atom cyclitol containing five hydroxyl groups in equatorial position and one in axial position. It is a substance essential for the growth and the well-being of cells since it is involved in important cellular pathways: it is the precursor of phosphatidylinositol , component of the biological membranes having a central role in the cellular signalling. It is broken down by phospholipase C into Diacylglycerol , involved in the adjustment of the family of protein kinase C, and inositol triphosphate, involved in several transcriptional pathways. It can be assumed with the diet, through the assumption of cereals, seeds and fruits or it can be synthetized starting from glucose. Nowadays, it is used in the prevention of the defects of the neural tube and in the syndrome of the polycystic ovary since it is known as being an insulin-sensitising agent. Inositol belongs to the vitamins of group B and it is also known as vitamin B7. The vitamins of group B favour the normal operation of the nervous system since they contribute in maintaining the normal levels of homocysteine; it is demonstrated that the excess in homocysteine is a potential risk factor of the cognitive decline and it linked to loss of attention and concentration. Therefore, vitamin B7 helps in overcoming the states of physical and psychical stress by safeguarding the health of the nervous cells. A diet low in vitamins of group B is associated to high levels of oxidative stress, homocysteine and neural inflammation. A clinical study randomized in double blind carried out on 39 healthy adults, aged between 30 and 65, investigated the effect that a supplementation with high doses of vitamins B has on the brain metabolism and on the markers of the oxidative stress. The participants were divided into two groups: placebo group (n = 20) and group (n = 19) treated with high doses of vitamins B (about 300 mg) and the treatment was performed for a period of six months. At the end of the six months of supplementation the plasmatic levels of homocysteine result reduced and the levels of brain markers of oxidative metabolism and of myelination (N-acetylaspartate, creatine and choline) instead result increased. These results suggest that the vitamins of group B reduce oxidative stress and inflammation by increasing the oxidative metabolism at the brain level and by favouring myelination.

COLA

The plants of genus Cola belong to the family of Malvaceae and include several species, thereamong the most known and used ones are Cola nitida and Cola acuminata. They are evergreen trees originally coming from Western Africa which can reach a height of 10 - 20 meters. Cola has always been used for its stimulating properties: for thousands of years it has been chewed by the populations of Western Africa and by travellers, in the long distances, in order to exploit its effect in reducing hunger, thirst and fatigue. Cola includes phenolic compounds having antioxidant action such as catechins and procyanidins . The tonic and stimulating effect is due to the presence of caffein and theobromine which produce a strong sensation of euphoria and well being, by increasing the levels of attention, concentration and physical energy. The effects of caffein on SNC are mainly related to its action on the receptors of adenosine Ai and A2, presynaptic receptors mediating the inhibition of transmission of the nervous stimuli. Caffein blocks adenosine receptors and increases the nervous transmission. Several studies highlighted the co-localization of adenosine receptors and of dopaminergic receptors in many brain areas, by demonstrating that caffein strengthens the post-synaptic transmission of dopamine.

A clinical study examined the caffein effect in keeping attention. The 12 participants, aged between 20 and 25, were subjected to an adaptation of Bourdon test for evaluating concentration and attention. Two sessions of experiments were performed differentiating only for the treatment (250 mg of caffein or placebo) and the measurements were performed by means of ERPs (event-related potentials) . The obtained results showed that, although no changes in behaviour were recorded, the treatment with caffein had positive effects on the front and parietal activity, by showing an increase in the excitatory stimuli with consequent fatigue decrease.

Another clinical study examined the cardio-pulmonary response, after 30 minutes of moderate exercise, after the assumption of Cola nitida. The 20 recruited healthy volunteers were divided into four groups:

Group A: check;

- Group B: 100 mg/kg of Cola nitida;

Group C: 200 mg/kg of Cola nitida;

Group D: 400 mg/kg of Cola nitida.

The cardio-pulmonary parameters were evaluated before and after exercise. The recorded results showed that the assumption of Cola nitida increases pressure in dose- depending manner, it stimulates the contraction of the cardiac muscle and increases the respiratory frequency more than check, by strengthening the physical performances. As designated previously the composition of the present invention is effective in the prevention and/or treatment of symptoms related to physical and/or mental tiredness and fatigue.

The present invention allows to obtain at the same time an antioxidant effect, a stimulating effect of the cognitive functions and a stimulating effect of the muscle functions. In particular, the present invention allows to obtain:

Antioxidant effect

- Tonic-energic effect

The present composition constitutes a valid support in the periods of intense stress thanks to its capability of alleviating both physical and mental fatigue and sustaining the attention levels, by means of the combined action of its active principles. Schisandra , thanks to its high content in polyphenols, exerts antioxidant action and strengthens and sustains the cholinergic system involved in learning processes and memory and in conducting neuro muscle stimuli. Noni is a very powerful antioxidant which increases the mental well-being, reduces the physical tiredness and increases resistance. Inositol, or vitamin B7, helps in fighting the states of physical and mental stress by safeguarding the nervous cells. The extracts of plants belonging to the genus Cola, thanks to the content in caffein and other metabolites, increase the levels of attention and concentration and increase the physical energy by stimulating the nervous transmission.

In a preferred embodiment the composition of the present invention is prepared as pharmaceutical dosage form comprising from 1 mg to 5000 mg of at least an extract of Schisandra chinensis, preferably from 10 mg to 1000 mg; from 1 mg to 5000 mg of at least an extract of Morinda citrifolia, preferably from 10 mg to 2000 mg; from 0.5 mg to 50000 mg of inositol, preferably from 5 to 2500 mg; and from 1 mg to 5000 mg of at least an extract of a plant belonging to the genus Cola, preferably from 10 mg to 1000 mg apart from the usual excipients used in the preparation of the selected dosage form.

The preferred administration route of the composition of the present invention is oral.

The preferred pharmaceutical dosage forms to this purpose are capsules, tablets, sachets, orosoluble sachets, solutions, suspensions, powders, orosoluble powders, granules, syrups, bottles, vials.

The following examples are provided by pure way of illustration and not for limitative purposes of the scope of the invention as defined by the enclosed claims.

Examples

EXAMPLE 1

Pharmaceutical form: vials for oral administration

EXAMPLE 2

Pharmaceutical form: powder for oral solution

EXAMPLE 3

Pharmaceutical form: tablet

EXAMPLE 4

Pharmaceutical form: capsule. EFFECTIVENESS TEST

The effectiveness of the composition, the present invention relates to, was evaluated according to experimental protocols known to the person expert in the art. In particular for the evaluation of the several activities of the composition according to the present invention in vitro and/or in vivo assays, known in the scientific literature, can be used.

All test types provide the use of experimental groups formed by the single active elements and by the union thereof as set forth by the present patent.

The synergic effect on the motor skills of the examined active elements can be tested by means of "Rotarod test", which evaluates the strength time and the holding force of the mice placed on a rotor set to a determined rotation speed.

The effect on the cognitive functions, learning and spatial memory can be evaluated in vivo on mice by means of:

- eight-arm radial maze test: the mouse is inserted at the centre of an eight-arm maze. In some of them a reward is placed and the mouse capabilities of storing the place in which there is food are evaluated;

- Y maze test: the animal is placed inside a maze having three identical arms in which he has free access. One evaluates the way in which the animal studies the maze structure ;

- Morris water maze test: it consists in inserting the mouse in a pool full of water and in evaluating the animal capability of detecting a platform hidden just above the water by exploiting the external stimuli.

For evaluating the antioxidant action of the present composition ex-vivo biochemical analyses can be performed on tissue homogenates after having induced stress or cognitive alteration in the animal; by means of spectrophotometric method the levels of Glutathione and MDA (Malondialdehyde, product of the lipidic peroxidation) are evaluated. Moreover, the plasmatic levels of SAR, LOOH and homocysteine (markers of the oxidative stress) and of N- acetylaspartate, creatine and choline (markers of the oxidative metabolism and of myelination) can be evaluated. In order to demonstrate in vivo the antioxidant effectiveness of the composition of the present invention, even assays result to be suitable which consist in determining in animals the levels of endogenous substances such as, for example, reduced glutathione, glutathione peroxidase, glutathione S-transferase, superoxide dismutase and catalase, after administration of the active elements of interest. Even the measurement of the lipidic peroxidation is a valid method for estimating the antioxidant activity of the present composition.

The antioxidant effectiveness of the composition, the present invention relates to, can be evaluated by means of in-vitro essays such as for example: DPPH test, radical scavenging activity on the nitric oxide or on the peroxynitrile radical, TEAC (Total radical-trapping antioxidant parameter), FRAP (Ferric reducing-antioxidant power) , HORAC (Hydroxyl radical averting capacity) , ORAC (Oxygen radical absorbance capacity) tests and the like.

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