DESCRIPTION FIELD OF THE INVENTION

The present invention relates to a food supplement comprising L-tryptophan and vitamins from the B vitamins group, to pharmaceutical compositions comprising said food supplement and to the production process of said food supplement.

Technical problem

In the food supplement of the present invention L-tryptophan is combined with vitamins from the B vitamins group which results in a synergistic effect of the composition to the central nervous system in the field of the treatment of depression and anxiety. Each substance acts individually but the specific combination of active ingredients and their ratios of the present invention results in their synergy.

BACKGROUND OF THE INVENTION

Today's pace of life and style, especially in large cities, more than ever, exposes us to stress that causes numerous negative effects to our body. In addition, more people have problems with food digestion, which also leads to various disorders, which in turn results in exhaustion and the constant feeling of fatigue. Also today's diet often leads to insufficient intake of necessary nutrients, especially minerals and vitamins.

Depression is the most common disease nowadays. Depression causes a disruption in the chemical and electrical processes in the brain that regulate mood. Usually this disorder is preceded by a long and severe stress, poor diet or lack of exercise, but can also occur without the stress, therefore, due to the innate tendency to depression. It is a disease that affects the entire body, but primarily affects mood by making it profoundly reduced. The mood is extremely important function of the human body, because it serves to maintain the desire to live. Due to the suffering that causes, depression is a disease that induces the highest number of suicides - two-thirds, and according to some research, even more. Billions are spent annualy worldwide on drugs for alleviation depression and anxiety. L Tryptophan is a biochemical precursor of serotonin (neurotransmitter of the central nervous system). The brain uses it (along with vitamins B6 and B3 and magnesium) for the production of serotonin. Serotonin is a hormone that transmits signals between nerve cells. Changes in serotonin levels in the brain significantly affects the process of mood regulation. In case of depression a decreased amount of serotonin is released from nerve cells in the brain. On the other hand, by increasing level of serotonin in the blood, i.e. its precursor L-tryptophan the antidepressant effect of serotonin can be enhanced. Thus, serotonin is involved in mood regulation, food intake control (affects the feeling of hunger and satiety), affects sleep and dreaming, feeling of excitement, and the perception of pain.

L-Tryptophan is an essential amino acid, which means it is essential for life but cannot be produced by our body and is therefore exclusively supplied by food intake. L-Tryptophan is found in most protein based food, especially in chocolate, oats, dried dates, milk, yogurt, cheese, red meat, eggs, fish, poultry, sesame, chickpeas, sunflower seeds, pumpkin seeds, algae spirulina, bananas and peanuts. After absorption from foods in the body L-tryptophan is converted to 5-hidroksitiptamin (5HTP) and then into serotonin.

Lack of tryptophan can occur in people with poor overall protein intake. Likewise, fructose absorption disorders and lactose intolerance may result in the improper absorption of tryptophan in the intestine, and consequently lead to a reduced level of tryptophan in the blood (Ledochowski M, Widner B, Murr C, Sperner-Unterweger B, Fuchs D "Fructose malabsorption is associated with decreased plasma tryptophan" Scand. J. Gastroenterol. 36 (4) (2001) 367-71). The lack of tryptophan in the diet increases the risk for deficiency of Vitamin B3, since L-tryptophan participates in the process of Vitamin B3 production in the body. This leads to an increased risk for development of diseases and health complications that are associated with a deficiency of this vitamin. Lack of L-tryptophan can lead to depression, insomnia, suicidal thoughts and the excessive need for carbohydrate intake.

The enhanced L-tryptophan intake, e.g. in the form of a dietary supplement may help to treat depression (it acts as a serotonin precursor, and amplifies the effect of classical treatment), sleep disorders (reduces the time required to fall asleep), anxiety, premenstrual dysphonic disorder (PMDD), smoking cessation, reduces sensitivity to pain, acts against depression, relieves migraine headaches, helps to reduce anxiety and tension, and take part in alleviating some symptoms of disorders caused by alcohol and for the treatment of alcoholism.

Vitamins from the B vitamins group (B complex vitamins) belong to the water soluble vitamins which play a significant role in the cell metabolism and to human health. Among these vitamins are usually included vitamin Bl (thiamin, suitably as hydrochloride or mononitrate), vitamin B2 (riboflavin, vitamin G, suitably as sodium riboflavin 5'-phosphate), vitamin B3 (niacin and nicotinamide, vitamin P, suitably as nicotinamide or nicotinic acid), vitamin B5 (pantothenic acid, suitably as sodium or calcium D-pantothenate), vitamin B6 (pyridoxine, suitably as hydrochloride or as 5'-phosphate), vitamin B7 (biotin, vitamin H, suitably as D-biotin) vitamin B9 (folic acid, vitamin M, suitably as pteroyl monoglutamic acid), vitamin B 12 (cobalamin, suitably as cyanocobalamin or hydroxocobalamin), choline, inositol, PABA (para-aminobenzoic acid) and so- called vitamin B 15 (pangamic acid), which in some countries is not recognized as a vitamin B group member.

The B vitamins are very important for the life and health and are essential for the metabolism of proteins, sugars and fats in the body. Therefore, absence of any of the B vitamins can lead to serious illness, and a complete lack of some of the B vitamins can cause death (disease beriberi: Bl deficiency, pellagra: a deficiency of vitamin B3, and pernicious anemia caused by vitamin deficiency B 12). Even a small deficiency can impair the overall balance causing a major disruption in the body. In addition, the B vitamins are essential for the production of hormones and neurotransmitters, and thus for the operation and health of nerves. Therefore, lack of them can cause a complete decomposition of the nerves, as is the case of vitamin B 1 deficiency. Due to the lack of neurotransmitters, hormones, and other ingredients the huge changes in mental health can occur causing major depression, anxiety, and even severe psychosis and dementia, such as is the case of a B l , B3 and B 12 vitamins deficiency.

It is well known that vitamins Bl, B2, B3, B6 and B 12 contribute to the normal functioning of the nervous system. Also that pantothenic acid (B5) contributes to normal mental abilities. Moreover, vitamins B2, B3, B5, B6, B9 and B 12 help in reducing fatigue and exhaustion.

Vitamin B3 is unique because it can be synthesized in the body from tryptophan amino acid (60 mg tryptophan = 1 mg niacin).

Inulin is a plant polymer (polysaccharide fructose), which serves as a carbohydrate storage which is, in various quantities, present in more than 36000 plant species. Significant quantities of inulin can be found in Jerusalem artichokes, chicory, dandelion, onion and garlic. Inulin has a lower caloric value than other carbohydrates. Since it is indigestible vegetable fiber it has a lower caloric value than other carbohydrates, and its decomposition starts only in the colon. Inulin is a prebiotic, which means that it stimulates the development of beneficial bacteria in the gut (Bifidobacteria and Lactobacilli acidophilus), which have various effects to the health. They eliminate harmful microorganisms of the intestinal flora (Clostridium sp., Salmonella sp., E. coli, Candida sp.), prevent intestinal infections, improve mineral absorption by 20% (especially Ca, Mg and Fe), produce vitamins from B vitamin group, reduce the production of liver toxins, strengthen the immune system and help in prevention of colon cancer. Inulin, therefore, normalizes digestion, revitalizes the intestinal flora disrupted by antibiotic treatment, and helps with constipation and intestinal infections. In' addition, inulin normalizes triglycerides and cholesterol level in the blood, thus preventing cardiovascular diseases and gradually reduces fat deposits in obese people. Degradation of inulin does not cause increase of insulin levels in the blood, which makes it an excellent dietary supplement for diabetics. Among the food supplements so far known in the art, there is no combination of L-tryptophan and vitamins from B vitamins group in a single food supplement as descibed by the present invention. Moreover, the specific combination of ingredients, as well as ratios of the individual components of the present invention so far have not been described in the art. Also, a production process of the food supplement of the present invention is not known in the prior art.

DETAILED DESCRIPTION OF THE INVENTION

The Objective

The objective of the present invention is to combine in a food supplement L-tryptophan with the vitamins from the B vitamins group which results in a synergistic effect of the composition on the central nervous system. The effect on the nervous system is particularly evident in the field of treating depression and anxiety. The specific combination of active ingredients and ratios of the individual components according to the present invention, results in a synergistic effect of each of the active ingredients which constitute this food supplement.

The additional objects and advantages of the invention will be shown in part in the description that follows, and in part will be found out through the application of invention.

Definitions

The "w%" abbreviation refers to a weight fraction of individual components in the mixture, expressed as a percentage of the weight of the individual component of the mixture in the total weight of the mixture.

The "r/min" abbreviation refers to the revolutions per minute. Embodiments of the Invention

The present invention relates to a food supplement comprising L-tryptophan and vitamins from B vitamins group, to pharmaceutical compositions comprising said food supplement and to the production process of said food supplement.

The present invention relates to a food supplement comprising from 35.00 to 85.00 w % L- tryptophan and from 0.50 to 10.00 w % vitamins from B vitamins group and, optionally, pharmaceutically acceptable excipents, to pharmaceutical compositions comprising said food supplement and to the production process of said food supplement. The present invention relates to a food supplement comprising from 60.00 to 85.00 w % L-tryptophan, from 0.50 to 10.00 w % vitamins from B vitamins group, from 10.00 to 60.00 w % inulin and, optionally, pharmaceutically acceptable excipents, to pharmaceutical compositions comprising said food supplement and to the production process of said food supplement.

In one suitable aspect, the present invention relates to a food supplement comprising 73.53 w % L- tryptophan and 4.05 w % vitamins from B vitamins group and, optionally, pharmaceutically acceptable excipents. In a further aspect, the present invention relates to a food supplement comprising 73.53 w % L-tryptophan, 4.05 w % vitamins from B vitamins group, 20.33 w % inulin and, optionally, pharmaceutically acceptable excipents. It will be apparent to one skilled in the art that the rest to the 100.00 w % are a pharmaceutically acceptable excipents.

In one aspect of the invention a food supplement comprises from 35.00 to 85.00 w % L-tryptophan. In a further aspect of the invention a food supplement comprises from 60.00 to 85.00 w % L- tryptophan. In a yet further aspect of the invention a food supplement comprises from 65.00 to 80.00 w % L-tryptophan. In even further aspect of the invention a food supplement comprises from 70.00 to 75.00 w % L-tryptophan. In a further suitable aspect of the invention a food supplement comprises 73.53 w % L-tryptophan. In a yet further suitable aspect of the invention a food supplement comprises 42.37 w % L-tryptophan.

In one aspect of the invention a food supplement comprises from 0.5 to 10 w % vitamins from B vitamins group. In a further aspect of the invention a food supplement comprises from 1.5 to 8 w % vitamins from B vitamins group. In a yet further aspect of the invention a food supplement comprises from 3 to 7 w % vitamins from B vitamins group. In even further aspect of the invention a food supplement comprises from 2 to 4.50 w % vitamins from B vitamins group. In a further suitable aspect of the invention a food supplement comprises 4.05 w % vitamins from B vitamins group. In a yet further suitable aspect of the invention a food supplement comprises 2.33 w % vitamins from B vitamins group.

In one aspect of the invention in the food supplement of the present invention vitamins from the B vitamins group are selected from Bl , B2, B3, B5, B6, B7, B9, B 12, choline, inositol, PABA and vitamin B 15. In a further aspect of the invention in the food supplement of the present invention vitamins from the B vitamins group are selected from B l , B2, B3, B5, B6, B9 and B 12.

In one aspect of the invention a food supplement comprises from 0.05 to 0.30 w % vitamin B l . In a further aspect of the invention a food supplement comprises from 0.10 to 0.25 w% vitamin Bl . In a yet further aspect of the invention a food supplement comprises from 0.15 to 0.20 w % vitamin B 1. In a further suitable aspect of the invention a food supplement comprises 0.19 w % vitamin B 1. In a yet further suitable aspect of the invention a food supplement comprises 0.1 1 w % vitamin Bl . In one aspect of the invention a food supplement comprises from 0.10 to 0.40 w % vitamin B2. In a further aspect of the invention a food supplement comprises from 0.15 to 0.35 w% vitamin B2. In a yet further aspect of the invention a food supplement comprises from 0.20 to 0.30 w % vitamin B2. In a further suitable aspect of the invention a food supplement comprises 0.25 w % vitamin B2. In a yet further suitable aspect of the invention a food supplement comprises 0.14 w % vitamin B2. In one aspect of the invention a food supplement comprises from 0.50 to 3.50 w % vitamin B3. In a further aspect of the invention a food supplement comprises from 1.00 to 3.00 w % vitamin B3. In a yet further aspect of the invention a food supplement comprises from 1.50 to 2.50 w % vitamin B3. In a further suitable aspect of the invention a food supplement comprises 1.96 w % vitamin B3. In a yet further suitable aspect of the invention a food supplement comprises 1.13 w % vitamin B3. In one aspect of the invention a food supplement comprises from 0.50 to 2.50 w % vitamin B5. In a further aspect of the invention a food supplement comprises from 0.55 to 2.00 w% vitamin B5. In a yet further aspect of the invention a food supplement comprises from 0.60 to 1.50 w % vitamin B5. In a further suitable aspect of the invention a food supplement comprises 0.99 w % vitamin B5. In a yet further suitable aspect of the invention a food supplement comprises 0.57 w % vitamin B5. In one aspect of the invention a food supplement comprises from 0.10 to 0.40 w % vitamin B6. In a further aspect of the invention a food supplement comprises from 0.15 to 0.35 w% vitamin B6. In a yet further aspect of the invention a food supplement comprises from 0.20 to 0.30 w % vitamin B6. In a further suitable aspect of the invention a food supplement comprises 0.25 w % vitamin B6. In a yet further suitable aspect of the invention a food supplement comprises 0.14 w % vi tamin B6. In one aspect of the invention a food supplement comprises from 0.01 to 0.06 w % vitamin B9. In a further aspect of the invention a food supplement comprises from 0.02 to 0.05 w% vitamin B9. In a yet further aspect of the invention a food supplement comprises from 0.02 to 0.03 w % vitamin B9. In a further suitable aspect of the invention a food supplement comprises 0.03 w % vitamin B9. In a yet further suitable aspect of the invention a food supplement comprises 0.02 w % vitamin B9. In one aspect of the invention a food supplement comprises from 0.20 to 0.55 w % vitamin B 12. In a further aspect of the invention a food supplement comprises from 0.30 to 0.50 w% vitamin B 12. In a yet further aspect of the invention a food supplement comprises from 0.35 to 0.45 w % vitamin B 12. In a further suitable aspect of the invention a food supplement comprises 0.38 w % vitamin B 12. In a yet further suitable aspect of the invention a food supplement comprises 0.22 w % vitamin B 12.

In one suitable aspect of the invention a food supplement comprises 73.53 w % L-tryptophan, 0.19 w % vitamin B l , 0.25 w % vitamin B2, 1.96 w % vitamin B3, 0.99 w % vitamin B5, 0.25 w % vitamin B6, 0.03 w % vitamin B9, 0.38 w % vitamin B 12 and, optionally, pharmaceutically acceptable excipients. In a further suitable aspect of the invention a food supplement comprises 73.53 w % L-tryptophan, 0.19 w % vitamin Bl , 0.25 w % vitamin B2, 1.96 w % vitamin B3, 0.99 w % vitamin B5, 0.25 w % vitamin B6, 0.03 w % vitamin B9, 0.38 w % vitamin B 12, 20.33 w % inulin and, optionally, pharmaceutically acceptable excipients. It will be apparent to one skilled in the art that the rest to the 100.00 w % are a pharmaceutically acceptable excipents.

In one aspect of the invention a food supplement comprises from 10.00 to 60.00 w % of inulin which serves as a filler. In a further aspect of the invention a food supplement comprises from 10.00 to 55.00 w % inulin. In a yet further aspect of the invention a food supplement comprises from 20.00 to 50.00 w % inulin. In even further aspect of the invention a food supplement comprises from 25.00 to 40.00 w % inulin. In a further suitable aspect of the invention a food supplement comprises 54.09 w % inulin. In a yet further suitable aspect of the invention a food supplement comprises 20.33 w % inulin.

A food supplement according to the invention is intended for oral administration, and the pharmaceutical dosage forms may be, for example capsules, tablets, granules, powders, emulsions and suspensions, all of which may be prepared using conventional methods of preparation. The conventional pharmaceutically acceptable excipients and adjuvants can be used, which are preferably selected from diluents and carriers such as cellulose powder, microcrystalline cellulose, cellulose derivatives such as hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose, dibasic calcium phosphate, corn starch, pre-gelatinized starch, polyvinyl pyrrolidone (Povidone) etc.; a lubricant such as stearic acid, magnesium stearate, sodium stearyl fumarate, glycerol tribehenate, etc.; disintegrants (eg. carboxymethylcellulose calcium), and the like; flow control agents such as colloidal silica, talc and the like; crystallization retarders such as Povidone, etc.; caking preventing agent such as magnesium stearate (preferably of plant origin), colloidal silicon dioxide and the like; solubilizers such as Pluronic, Povidone, etc.; coloring agents, including dyes and pigments such as red or yellow iron oxide, titanium dioxide, talc and the like; pH control agents (buffer substances) such as citric acid, tartaric acid, fumaric acid, sodium citrate (mono-, di-and tri-), dibasic calcium phosphate, dibasic sodium phosphate and the like; surfactants and emulsifiers such as Pluronic, polyethylene glycols, sodium carboxymethyl cellulose, polietioksilirano and hydroxylated castor oil and the like; and mixtures of two or more of these excipients and / or adjuvants.

In one aspect of the invention a food supplement according to the present invention is intended for oral administration. In a further aspect of the invention a food supplement according to the present invention is formulated for oral administration as a capsule. Suitably, capsule is size "0", and the average weight of a full capsule is 300 mg, which includes the weight of the food supplement and weight of the capsule. More suitably, capsule is size "1", and the average weight of a full capsule is 280 mg, which includes the weight of the food supplement and weight of the capsule. In one aspect of the invention single capsule contains between 100 mg to 300 mg L-tryptophan and vitamins from B vitamins group alltogeher. In a further aspect of the invention single capsule contains between 150 mg to 250 mg L-tryptophan and vitamins from B vitamins group alltogeher. In yet further suitable aspect of the invention single capsule contains 157.03 mg L-tryptophan and vitamins from B vitamins group alltogeher. Suitably, single capsule which contains 157.03 mg L- tryptophan and vitamins from B vitamins group comprises repsectivelly 150 mg L-tryptophan, 0.33 mg vitamin B l , 0.42 mg vitamin B2, 4 mg vitamin B3, 1.8 mg vitamin B5, 0.42 mg vitamin B6, 60 μg vitamin B9 and 0.75 μg vitamin B 12, and inulin and pharmaceutically acceptable excipients. The dose of the food supplement according to the present invention is suitable for adults over 12 years of age and can vary from 600 to 2400 mg per day of L-tryptophan and vitamins of B vitamins group in total, in single or several divided portions, and can be suitably increased or reduced depending on the age, body weight and medical condition. In one aspect of the invention the dose of the food supplement is between 900 to 1200 mg per day of L-tryptophan and vitamins of B vitamins group in total, in a single or several devided portions. In a suitable embodiment of the invention the daily dose is 3 to 4 times per two capsules a day. The recommended daily dose is 6-8 capsules which contain:

In one aspect of the invention a process for production of the food supplement of the invention comprises the steps of:

i) Phase I: Preparation of the 5% premix of folic acid,

ii) Phase II: Preparation of the vitamin B group premix,

iii) Phase III: Preparation of the food supplement final mixture, and

iv) Phase IV: Preparation of the final pharamaceutical composition. In a further aspect of the invention a process for production of the food supplement of the invention comprises the steps of:

i) Phase I: Preparation of the 5% premix of folic acid which comprises adding weighed amounts of inulin and folic acid into the tank for homogenization of powders by stirring; ii) Phase II: Preparation of the vitamin B group premix which comprises adding weighed amounts of inulin, colloidal silicon dioxide, calcium D-(+)-pantothenate, vitamin B12 triturate (0.1%), the 5% premix of folic acid from Phase I, sodium riboflavin-5 '-phosphate, thiamine hydrochloride, and pyridoxine hydrochloride into the tank for homogenization of powders by stirring, homogenization by stirring, grinding in a mill with a sieve, returning to the tank for homogenization, and additional homogenization by stirring;

iii) Phase III: Preparation of the food supplement final mixture which comprises adding weighed amounts of L-tryptophan, inulin, vitamin B group premix from Phase II, and nicotinamide into the tank for homogenization of powders by stirring, homogenization by stirring, adding pre- weighed magnesium stearate, and additional homogenization by stirring; and_

iv) Phase IV: Preparation of the final pharamaceutical composition from the food supplement final mixture prepared in Phase III using any of the conventional methods for preparation of the pharmaceutical composition chosen among capsules, tablets, granules, powders, emulsions, and suspensions.

In yet further aspect of the invention a process for production of the food supplement of the invention comprises the steps of:

i) Phase I: Preparation of the 5% premix of folic acid which comprises adding weighed amounts of inulin and folic acid into the tank for homogenization of powders by stirring, homogenization by stirring at 10-40 r/min, sutably at 12-20 r/min during 10-60 minutes, suitably during 20 minutes;

ii) Phase II: Preparation of the vitamin B group premix which comprises adding weighed amounts of inulin, colloidal silicon dioxide, calcium D-(+)-pantothenate, vitamin B 12 triturate (0.1%), the 5% premix of folic acid from Phase I, sodium riboflavin-5'-phosphate, thiamine hydrochloride, and pyridoxine hydrochloride into the tank for homogenization of powders by stirring, homogenization by stirring at 10-40 r/min, suitably at 12-20 r/min, during 2 to 10 minutes, suitably during 3 minutes, grinding in a mill with a sieve, suitably using 1 mm sieve returning to the tank for homogenization, and additional homogenization by stirring at 10-40 r/min, suitably at 12-20 r/min during 10-60 minutes, suitably during 20 minutes;

iii) Phase III: Preparation of the food supplement final mixture which comprises adding weighed amounts of L-tryptophan, inulin, vitamin B group premix from Phase II, and nicotinamide into the tank for homogenization of powders by stirring, homogenization by stirring at 10-40 r/min, suitably at 12-20 r/min during 10-60 minutes, suitably during 20 minutes, adding pre-weighed magnesium stearate, and additional homogenization by stirring at 10-40 r/min, suitably at 12-20 r/min, during 2 to 10 minutes, suitably during 3 minutes; and

iv) Phase IV: Preparation of the final pharamaceutical composition from the food supplement final mixture prepared in Phase III using any of the conventional methods for preparation of the pharmaceutical composition chosen among capsules, tablets, granules, powders, emulsions, and suspensions. In one suitable aspect of the invention the final pharmaceutical composition is capsule, thus the mixture prepared in Phase III is capsulated, and filled capsules are dusted off before being placed in bottles, which are then closed, labeled and placed into a cardboard box. In a further aspect of the present invention in each capsule, in addition to the final mixture of the food supplement from a phase III, the additional amount of inulin is added, suitably 150 mg of inulin. In a yet further aspect of the present invention to the the food supplement final mixture prepared in Phase III is first added the additional amount of inulin, suitably 735.30 mg, the resulting mixture is homogenized and optionally sieved, and then encapsulated.

In one aspect the present invention relates to a food supplement prepared according to a process which comprises the steps of:

i) Phase I: Preparation of the 5% premix of folic acid,

ii) Phase II: Preparation of the vitamin B group premix,

iii) Phase III: Preparation of the food supplement final mixture, and

iv) Phase IV: Preparation of the final pharamaceutical composition.

The illustration of the present invention, is provided by following, non-limiting example.

EXAMPLES

All used ingredients are commercially available. Some ingredients may contain 0.5 to 10% w% of moisture and the moisture content is included in the sample weight of each of the phases of the preparation procedure.

In the 0.1% vitamin B 12 trituate maltodextrin may be used as a carrier, and as buffering agents (stabilizers of vitamin B 12) trisodium citrate and citric acid. Example 1.

Phase I: Preparation of the 5% premix of folic acid on the scale of 6.43 g

Procedure: Weighed quantities of inulin and folic acid were added to the homogenizing tank (V = 20 - 25 mL) and homogenized by stirring at 12-20 r/min for 20 minutes, without indication of agglomeration, to give a fine yellow powder.

Faza II: Preparation of the vitamin B group premix on the sacale of 41.18 g

Procedure: The weighed amounts of inulin, colloidal silicon dioxide, calcium D-(+)-pantothenate, 0.1% vitamin B12 triturate, the 5% premix of folic acid from Phase I, sodium riboflavin-5'- phosphate, thiamine hydrochloride, and pyridoxine hydrochloride were added into the tank for homogenization (V= 200-250 mL), the obtained mixture was stirred at 12 r/min during 3 minutes; then ground in a mill and sieved (1 mm sieve); then quantitatively transferred back to the homogenization tank where it was homogenized by stirring at 12-20 r/min during 20 minutes to give a fine orange powder.

Phase III: Preparation of the food supplement final mixture on the sacale of 1 kg

Procedure: The weighed amounts L-tryptophan, inulin, nicotinamide (vitamin B3) and vitamin B group premix from Phase II were added into the tank for homogenization of powders (V= 4-5 L) where mixture was homogenized by stirring at 12 r/min during 20 minutes, then magnesium stearate was added and mixture additionally homogenized by stirring at 12 r/min during 3 minutes to give the final mixture of the food supplement which was used for preparation of final pharmaceutical composition in Phase IV. Phase IV: Preparation of the final pharamaceutical composition - capsule

Metod A: Procedure: The final mixture of the food supplement from Phase III (lOOO.OOg) was encapsulated to "0" size, white vegetable (HPMC) capsules (Vcaps Plus, Capsugel) giving the average weight of filled capsules of 300 mg. Encapsulation gave 4902 filled capsules. The filled capsules were dusted off before being placed in bottles, which were then closed, labeled and placed into a cardboard box.

1. Theoretical weight of the capsule: 300 mg ± 15% (255-345 mg)

2. Weight of capsule contents: 204 mg ± 15% (173,4-234,6 mg)

3. Mass of empty capsule: 96 mg ± 6 mg

4. Appearance: white capsule

5. Length: 21 ,7 mm ± 0,7 mm (21 ,0-22,4 mm)

6. The appearance of the capsule contents: fine powder.

Metod B: Procedure: same as that of Method A in Phase IV, except that in each capsule the additional amount of 150 mg of inulin was addec 1 which serves as a filler.

Metod C: Procedure: The final mixture of the food supplement from Phase III (lOOO.OOg) and an additional amount of inulin (735.30 mg) was dispensed into the tank for homogenization of powders (V L = 4-5) where mixture was first homogenized by stirring at 12 r/min for 20 minutes, optionally sieved (1 mm sieve) and further homogenized by stirring, and then the thus obtained mixture of the food supplement filled in a capsules as described in Method A.