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  Pharmaceutical Patents  

 

Title:  Method for selectively inhibiting reuptake of serotonin and norepinephrine using yeast extract
United States Patent: 
7,473,426
Issued: 
January 6, 2009

Inventors:
 Choe; Yun Seok (Ilsan3-dong, Ilsan-gu, Goyang-city, Kyungki-do, KR), Kang; Il Jun (Chuncheon, KR), Suh; Hyung Joo (Seoul, KR), Choi; Young Chun (Chuncheon, KR), Yun; Hee Sun (Seoul, KR), Kim; Kyung Mi (Seoul, KR), Ahn; Sang Wook (Seoul, KR), Kim; Ki Won (Cheonju, KR), Hwang; Won Jun (Incheon, KR), Kim; Jin Man (Seoul, KR)
Assignee:
  Choe; Yun Seok (KR), Lee; Moon Sup (KR)
Appl. No.:
  11/141,497
Filed:
 May 31, 2005


 

George Washington University's Healthcare MBA


Abstract

A yeast extract or yeast-derived bioactive peptide having an activity of selectively inhibiting reuptake of serotonin and norepinephrine, which can be effectively used in preventing or treating various diseases related to reuptake of serotonin and norepinephrine, especially depression, anxiety, stress, fatigue and obesity is provided. Also, a yeast extract or yeast-derived bioactive peptide having activities as an anti-stress agent, an anti-fatigue agent, premenstrual syndrome (PMS) and menstrual pain relaxants, and a brain-neurotrophic factor, and a method for preparing the bioactive peptide is provided. The yeast extract or yeast-derived bioactive peptide is effective in relieving stress, nervousness, anxiety, tension, insomnia, fatigue, and imbalance in the autonomic nerve regulation. Therefore, the yeast extract or yeast-derived bioactive peptide is available as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, a brain-neurotrophin, and a source of active foods having these activities.

Description of the Invention

SUMMARY OF THE INVENTION

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as an selective serotonin and norepinephrine reuptake inhibitor.

The present invention provides a new use of yeast and bioactive peptides obtained by hydrolyzing yeast extract as an anti-depression agent to treat depression caused from undue reuptake of serotonin and norepinephrine.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as an anti-anxiety agent to treat anxiety caused from undue reuptake of serotonin and norepinephrine.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as an anti-stress agent to treat stress caused from undue reuptake of serotonin and norepinephrine.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as an anti-fatigue agent to recover fatigue caused from stress or muscular fatigue caused from exercises.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as an anti-obesity agent to treat obesity caused from undue reuptake of serotonin and norepinephrine.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as a natural brain-neurotrophin to treat stress disorders and maintain balance in the human autonomic nervous system.

The present invention provides a new use of yeast extract and bioactive peptides obtained by hydrolyzing yeast as premenstrual syndrome and menstrual pain relaxants to reduce premenstrual syndromes (PMS) and menstrual pains similar to symptoms from stress and chronic fatigue.

The present invention provides a functional beverage, functional gum or functional edible film containing a yeast extract or bioactive peptides obtained by hydrolyzing yeast as an active component having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, anti-obesity agent, premenstrual syndrome and menstrual pain relaxants and a sleeping aids to treat neurogenic disorder, especially imbalance in the autonomic nervous system, caused from stress.

In accordance with an aspect of the present invention, there is provided a method for selectively inhibiting reuptake of serotonin and norepinephrine comprising administering an effective amount of a yeast extract. In the present invention, the term "yeast extract" is understood to include a crude yeast extract, a purified yeast extract and bioactive peptides obtained by hydrolyzing yeast.

In the present method, the yeast extract may be prepared using conventional extraction methods, for example by culturing yeasts, recovering the cultured yeast with centrifugation and hydrolyzing the recovered yeast. Preferably, the hydrolyzing step can be carried out by autolyzing the recovered yeast at a temperature of about 35-70.degree. C. and/or hydrolyzing with a protease.

In the present method, the yeast extract may be used for preventing or treating a disease related to reuptake of serotonin and norepinephrine. In the present invention, the disease related to reuptake of serotonin and norepinephrine include any disease which is induced by deficiency or hypofunction of serotonin and norepinephrine caused from undue reuptake of them, for example depression, anxiety, stress, fatigue, obesity and the like.

In accordance with an aspect of the present invention, there is provided a method for reduce or relax premenstrual syndromes (PMS) and menstrual pains.

In the present method, the yeast extract is administered in form of a pharmaceutical composition or a functional food comprising the yeast extract as an active component, preferably a functional beverage, a functional gum or functional edible film.

DETAILED DESCRIPTION OF THE INVENTION

Hereinafter, the present invention will be described in greater detail.

In accordance with an aspect of the present invention, there is provided a yeast extract derived from yeast and having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor.

In the yeast extract according to the present invention, the source yeast contains excess high-quality proteins, minerals, and B vitamins. The yeast extract is obtained by known general extractions methods, for example, using an autolytic enzyme or protease. Commercially available yeast extracts were found to have activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, PMS and menstrual pain relaxants, and a brain-neurotrophic factor, like the yeast extract according to the present invention.

The yeast extract according to the preset invention having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, is prepared by autolyzing the yeast preferably at a temperature of about 35-70.degree. C., and more preferably, a temperature of about 50-60.degree. C.

Only with the autolysis at a high temperature greater than or equal to about 35.degree. C., an effective anti-stress activity can be induced to the yeast extract. It is believed that heating the yeast simultaneously triggers the release of stress-resistant substances to provide an anti-stress effect during autolysis. Alternatively, by applying an additional stress such as ultrasonic waves or vibrations, a new strain of yeast with enhanced stress resistance can be screened. The anti-stress activity of the yeast extract according to the present invention can be further improved by repeatedly applying such stresses to enhance the release of stress-resistant substances.

In the yeast extract according to the preset invention having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, it is preferable that the yeast is hydrolyzed with a protease during or after the autolysis. According to this invention, only a supernatant obtained by centrifuging hydrolytes produced in the hydrolysis may be included in the yeast extract according to the present invention having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor.

Also, the present invention provides a yeast-derived peptide having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, the yeast-derived peptide characterized by including molecules of 10,000 daltons or less obtained by ultrafiltrating the supernatant from the centrifugation performed in the above preparation of the yeast extract according to the present invention.

In the yeast-derived peptide according to the present invention, the molecular weight cutoff value of 10,000 was determined by considering a variety of activities of peptides as an active food source to regulate body functions (New Technology Trend Report of 2000, Active Food, Korean Industrial Property Office). In general, molecules of a molecular weight of 10,000 or less are called "peptides". The brain has a brain blood barrier which blocks macromolecules of a molecular weight of 15,000 or greater to protect the cerebrovascular system. Therefore, the yeast-derived peptide according to the present invention having a molecular weight of 10,000 or less can easily pass through the brain blood barrier.

Alternatively, a yeast-derived peptide according to the present invention having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor is characterized by comprising glutamic acid and aspartic acid of about 15-25 mol % each.

Since the yeast-derived peptide according to the present invention is rich in glutamic acid and aspartic acid, which are amino acids involved in the synthesis of muscarinic acetylcholine receptors, it is effective in alleviating stress disorders caused by the imbalance in the autonomic nervous system, including PMS and menstrual pains.

In accordance with another aspect of the present invention, there is provided a method for preparing a yeast extract having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, the method comprising: incubating a strain of yeast until a maximum growth phase; inducing autolysis of the yeast at a temperature of about 35-70.degree. C. and simultaneously hydrolyzing the yeast with an addition of a protease; and attaining a supernatant by centrifuging the hydrolytes from the yeast.

Alternatively, the method for preparing the yeast extract according to the present invention may further comprise screening a new strain of the yeast survived resisting to stress by heating at high-temperature, applying ultrasonic waves and vibrations, and changing pH to a degree not to cause destruction before the incubation until the maximum proliferation stage.

Alternatively, the method for preparing the yeast extract according to the present invention may further comprise inducing secretion of metabolites resistant to stress by applying a physical or chemical stress selected from the group consisting of high-temperature heating, ultrasonic waves, vibrations, and pH variations, to a degree not to cause destruction before the autolysis.

This is based on the fact all organisms exhibit an alarm reaction when a stress is perceived for the first time and actively resists against the stress if the stress is not relieved to induce physiological changes for homeostasis (Seyle, 1956). In other words, when yeast is subjected to stress, it secretes a variety of enzymes to resist against the stress. The released enzymes are low molecular weight proteins having an activity as a neurotrophic factor to relieve the human body of the stress. Based on this fact, the yeast extract preparation method according to the present invention has been realized.

Physical or chemical stimuli applied to yeast in the preparation of a yeast extract according to the present invention to induce the generation and release of anti-stress substances from the yeast include heating at a temperature of about 35-45.degree. C. which is higher than the optimum growth temperature of the yeast, ultrasonic waves, vibrations, and pH variations, to a degree not to destroying the yeast.

Preferably, the method for preparing the yeast extract according to the present invention comprises: screening a new strain of yeast resistant to stress by incubating the yeast with applications of ultrasonic waves and vibrations; inoculating the screened strain of the yeast on YM medium, incubating the medium at a temperature of about 22-25.degree. C. until its exponential growth phase, collecting the yeast cells by centrifugation, and diluting the collected cells with a 1%-peptone buffer; applying ultrasonic waves and vibrations to the dilute at a high temperature of about 33-45.degree. C. as stresses for 8 hours to induce the generation and release of stress-resistant metabolites from the yeast; autolysing the yeast product at a temperature of about 50-60.degree. C. and simultaneously hydrolyzing the yeast product with the addition of a protease; and centrifuging hydrolytes to obtain a supernatant.

In accordance with another aspect of the present invention, there is provided a method for preparing a yeast-derived peptide having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, the method characterized by comprising obtaining peptides only having a molecular weight of 10,000 daltons or less by ultrafiltrating the supernatant from the centrifugation in the preparation of the yeast extract according to the present invention.

According to the present invention, only low molecular weight yeast-derived peptides of 10,000 or less is selected by ultrafiltrating the supernatant from the centrifugation described above through a membrane having a molecular weight cutoff value of 10,000. As a result, substances that would drop the efficacy of the yeast extract are removed while the low molecular weight substances capable of easily being absorbed into the body and capable of passing through the brain blood barrier with high bioavailability are separated and purified.

Alternatively, the method for preparing the yeast extract or the yeast-derived peptide according to the present invention may further involve drying and grinding the yeast extract or the yeast-derive peptide.

In accordance with another aspect of the present invention, there is provided an anti-stress agent comprising the yeast extract or the yeast-derived peptide prepared by any of the above-described methods according to the present invention as an active component.

The anti-stress agent according to the present invention has an activity as a tranquilizer, a relaxant, an anti-anxiety agent, or a sleeping aids, and is effective in alleviating PMS and menstrual pains.

In accordance with another aspect of the present invention, there is provided an anti-fatigue agent (fatigue recovering agent) comprising the yeast extract or the yeast-derived peptide prepared by any of the above-described methods according to the present invention as an active component.

In accordance with another aspect of the present invention, there is provided a neurotrophin (neurotrophic factor) comprising the yeast extract or the yeast-derived peptide prepared by any of the above-described methods according to the present invention as an active component.

The present invention also provide a PMS and menstrual pain relaxant composition comprising the yeast extract or the yeast-derived peptide prepared by any of the above-described methods according to the present invention as an active component.

Preferably, the PMS and menstrual pain relaxant composition according to the present invention comprises: about 10-90% by weight dried powder of the yeast extract or the yeast-derived peptide; about 5-80% by weight chitosan; and about 5-80% by weight herbal powder of 5-80% by weight, based on the total weight of the premenstrual syndrome and menstrual pain relaxant composition. In this case, the chitosan may be a water-soluble macromolecule of a molecular weight greater than or equal to about 300,000. Preferably, the herbal powder is derived from at least one selected from the group consisting of Korean angelica root, Salviae Radix, Curcuma aromatica, Zedoariae Rhizoma, mint, liquorice, ginger, Gastrodia, white Atractylis, Cnidium officinale, cinnamon, and ginseng.

The functional food according to present invention can be prepared by using conventional method well known to food industry. For example, the functional food can be prepared in the forms of tablet, capsule, powder, granule, solution, pellet and the like. Preferably, the functional food can be prepared in the forms of functional beverage, functional gum and edible films.

In accordance with another aspect of the present invention, there is provided an active beverage having activities as an anti-depression agent, an anti-anxiety agent, an anti-stress agent, an anti-fatigue agent, an anti-obesity agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, the active beverage comprising the yeast extract or the yeast-derived peptide prepared by any of the above-described methods according to the present invention.

The yeast extract or yeast-derived peptide contained in the active beverage according to the present invention contains anti-stress substances released by applying stress to yeast recognized as safe. Therefore, the active beverage prepared using the yeast extract or yeast-derive peptide according to the present invention provides an anti-stress effect as a tranquilizer, a relaxant, a sleeping drug, etc., and can be conveniently taken without concern about any side effect.

Preferably, the active beverage according to the present invention comprises: about 0.1-10% by weight the yeast extract or the yeast-derived peptide by any of the above-described methods according to the present invention; about 10-25% by weight common additives for beverage including a sweeter and an acidulant, based on the total weight of the bioactive beverage; and the balance water.

In the present invention, the common additives for beverage include liquid fructose, sucrose, maltodextrin, glucose, citric acid, nicotinamide, pantothenic acid, sodium benzoate, and kinds of flavors. Preferably, in the preparation of the bioactive beverage according to the present invention, any fruit juice, for example, ume juice, is added.

The present invention also provides a method for preparing an bioactive beverage having activities as an anti-stress agent, an anti-fatigue agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, the method comprising: incubating a strain of yeast until a maximum growth phase; inducing autolysis of the yeast at a temperature of about 35-70.degree. C. and simultaneously hydrolyzing the yeast with an addition of a protease; attaining a supernatant by centrifuging hydrolytes produced in the hydrolysis; mixing the supernatant with active carbon in water and sterilizing the mixture under pressure; and purifying the sterilized mixture until it loses its color by filtering the sterilized mixture with suction.

In preparing a beverage with the yeast extract containing anti-stress substances, purification is necessary to remove the yeast odor. For the purification, after mixing the prepared yeast extract with active carbon and water, the mixture is sterilized under pressure and filtered. Active carbon almost not adsorbs amino acid (Hyung-ik Song and Jung-yeub Shin, Contemporary Fermentation Engineering, 1998, 293). Based on this nature of active carbon, the yeast extract was mixed with active carbon for decolorization and deodorization in the present invention.

The active beverage according to the present invention having activities as an anti-stress agent, an anti-fatigue agent, premenstrual syndrome and menstrual pain relaxants, and a neurotrophic factor, is prepared by diluting the beverage concentrate after the filtering. An bioactive beverage according to the present invention can be prepared by any general beverage preparation method without limitation, as long as the yeast extract prepared by any of the methods described above according to the present invention is incorporated therein.

Preferably, the active beverage preparation method according to the present invention further comprises: screening a new strain of yeast survived resisting to stress by heating at high-temperature, applying ultrasonic waves and vibrations, and changing pH during incubation to a degree not to causing destruction; and inducing secretion of metabolites resistant to stress by applying a physical or chemical stress selected from the group consisting of high-temperature heating, ultrasonic waves, vibrations, and pH variations, to a degree not to causing destruction.

In the preparation of the bioactive beverage according to the present invention, it is preferable that the autolysis is performed at a temperature of about 50-60.degree. C., and the pressure sterilization is performed at about 1.5 atm and a temperature of about 100-125.degree. C. for about 10-15 minutes.

The functional gum according to the present invention can be prepared by conventional methods for preparing chewing gum, preferably by mixing and dissolving 50-65% by weight saccharide, 10-25% by weight sugar alcohol, 0.05-0.5% by weight citric acid, 0.01-0.05% by weight aspartame and 0.1-10% by weight the present yeast extract, adding and dissolving 20-40% by weight chewing base, adding and mixing 1-2% by weight fruits or herb fragrant powder, and heating and rolling to cast a chewing gum.

The edible film according to the present invention can be prepared by conventional methods for preparing edible film, preferably by preparing aqueous solution of polyhydric alcohol comprising polyhydric alcohol and water in a range of 0.2:99.8 to 20:80 weight ratio, dissolving water-soluble polysaccharide such as starch or pulluran and the present yeast extract, casting the resulting solution into a film, and drying the film to 25% by weight or less of water content. The edible film of the present invention can be heat-seamed and dissolved in hot bath, so can be used as materials of various food packaging bag.

To recover the human organs subjected to sustained stimulation of the sympathetic part due to stress, which causes imbalance in the autonomic nervous system, the parasympathetic part needs to be actively stimulated. To this end, there is a need to accelerate release of the neurotransmitter acetylcholine at synapses. In addition, acetylcholine receptors are necessary to efficiently deliver the released acetylcholine to neurons. Acetylcholine is bound to muscarinic receptors, as shown in FIG. 2 (see Original Patent). The presence of more muscarinic receptors in the synapsis results in more efficient transmission of acetylcholine. Therefore, more muscarinic receptors need to be synthesized. The muscarinic receptors are composed of seven fractions of protein and are believed to have a negatively charged site to couple with quaternary ammonium salt of acetylcholine significant for the muscarinic activity. The negatively charged site is considered to be derived from acidic amino acid residues, such as aspartic acid and glutamic acid (Keun-il Kang, 1992, Introduction to Medicinal Chemistry)

Therefore, peptide drugs enriched with acidic amino acids, aspartic acid and glutamic acid, to be used as a source material for the muscarinic acetylcholine receptors facilitates the biosynthesis of the muscarinic receptors in the human body and affects efficient transmission of acethylcholine of the parasympathetic part, thereby recover the imbalance in the autonomic nervous system due to the excess stimulation of the sympathetic part. As a result, the pathological conditions, both physical and emotional, caused from stress can be fundamentally inhibited or relieved.

The inventor has found that yeast extract is rich in amino acids effective in synthesizing the muscarinic receptors described above and can be effective in treating stress disorders caused from the imbalance in the nervous system.

A yeast extract according to the present invention is found to contain 18 kinds of amino acids with a great amount of aspartic acid and glutamic acid, which will be described later in Example 4. Aspartic acid and glutamic acid have a negatively charged R group. In other words, the result means that the yeast extract containing a large amount of negatively charged amino acids, which are essential to synthesis muscarinic receptors in the body, is a good source for the biosynthesis of the neurotransmitter acetylcholine distributed in the nervous system, including the parasympathetic part. Since the yeast extract according to the present invention accelerates transmission of the neurotransmitter at the parasympathetic part when the sympathetic part is excessively stimulated due to stress, the yeast extract according to the present invention has an activity as an anti-stress agent. Furthermore, the yeast extract according to the present invention can effectively relax premenstrual syndrome (PMS) and menstrual pains, which are similar to the symptoms from stress.

Food-grade yeast, generally recognized as safe (GRAS), contains 50% or greater quality protein, a large amount of minerals, and vitamin B complex. The yeast is rich in peptides as a source for the synthesis of neurotransmitters or their receptors, as described above. In particular, vitamin B and minerals contained in yeast are involved in general energy metabolism and activating the brain and neurons. Vitamin B6 (pyridoxine) is an essential nutrient for synthesizing neurotransmitters, including serotonin, dopamine, norepinephrine, gamma-amino butyric acid (GABA), and taurine. A deficiency of vitamine B6 causes dysthesia, anxiety, reduction in concentration, and hypomnesis. A deficiency of Vitamin B1 (thiamine) causes Korsakoff's syndromes such as weakness of memory, anaesthesia, and dementia. Vitamin B2 (riboflavin) acts as a coenzyme for the brain growth to be considered neurologically as an important neurotrophic factor.

In women with PMS or incapacitating menstrual pains, signs of progesterone deficiency, estrogen/progesterone imbalance, salt/water retention, prostaglandin deficiency or excess, prolactine excess, vitamin B6 (pyridoxine) deficiency, hypoglycemia, serotonin deficiency, etc. are observed. However, since the yeast extract according to the present invention contains a variety of amino acids, excess B vitamins, including B6, and excess minerals such as calcium and magnesium, hormone balance and metabolism activation are accelerated as those components are absorbed into the body, thereby rapidly reduce PMS and menstrual pains.

Furthermore, yeast contains comparatively excess selenium, which is essential to grow cranial nerve cells and to generate antioxidases. Deficiency of selenium results in depression and anxiety due to encephalopathy. Mineral enriched yeast, as an easily absorbable mineral supplement in the form of organically bound, chelated with minerals, such as selenium and chromium, is disclosed (U.S. Pat. No. 4,530,846 issued on Jul. 23, 1985, entitled "Method for the Production of Selenium Yeast"). In view of the description above, yeast is considered to be applicable as a bioactive food material and natural peptide source in the form of yeast extract or yeast-derived peptides with an activity as a good brain-neurotrophic factor capable of relaxing stress, anxiety, excitation, somnipathy due to stress, and fatigue.

In the present invention, a variety of excess exoenzymes produced are secreted through the cell wall of the yeast. Therefore, the exoenzymes are low molecular weight proteins, which are enough small to pass the yeast cell wall. Therefore, the exoenzymes can act as a brain-neurotrophic factor capable of easily passing through the brain blood barrier in the human body.

When the growth conditions, such as temperature, pH, and nutrient composition and amount, are extremely undesirable, yeast is autolysed by its own enzyme to hydrolyze the cytoplasm and subsequently destruct the cell wall so that the hydrolyzed cytoplasm comes out of the cell. Due to the yeast autolysis, bioavailability of the yeast can be enhanced. By autolysing yeast after generation of excess low molecular weight anti-stress substances in the yeast, yeast-derived peptides containing a variety of proteins, minerals, and B vitamins, which are extracted from the yeast by the autolysis and act as effective neurotrophic factors, can be obtained.

The effect of yeast extract in relaxing stress and fatigue as a neurotrophic factor has been described above. Hereinafter, the effect of relaxing PMS and menstrual pains will be described in greater detail.

First, the yeast extract contains excess choline as a complex with vitamin B. Choline is a precursor of the relaxant neurotransmitter acetylcholine. Therefore, muscular tension and pain due to uterine contraction during menstruation can be alleviated by increasing choline intake.

Choline exists in a variety of forms. Among many types of choline, salicylic choline has a pain alleviating effect similar to aspirin and thus may be effective in alleviating menstrual pains. Although the cause of menstrual pains has not been identified accurately, it is expected due to excess secretion of prostaglandin during menstruation, which is observed in women with menstrual pains. Analgesia such as acetylsalicylic acid (aspirin) or ibuprofen inhibits the synthesis of prostaglandin to reduce prostaglandin secretion. As a result, neurotransmission of the pain is inhibited to be less painful.

Second, vitamin B6 in the yeast extract composition is very important in the synthesis and metabolism of amino acids and proteins, functions as a cofactor for the synthesis of red blood cells and antibodies, and is involved in the synthesis of a variety of neurotransmitters. Deficiency of vitamin B6 results in anemia, dermatitis, neuropathy, and cramp. Also, vitamin B6 is known to be essential to relieve PMS and menstrual pains.

Third, niacin in the yeast extract composition is involved in the enzymatic reaction for the synthesis of coenzymes, NAD (nicotinamide adenine dinucleotide) and NADP (nicotinamide adenine dinucleotide phosphate). These coenzymes are known to accelerate energy generation in the intracellular mitochondria and to activate the function of brain cells to treat schizophrenia. Accordingly, niacin can be effectively used to alleviate the psychological syndromes of PMS.

Niacin is one of metabolites of the essential amino acid tryptophan. Tryptophan is a precursor of serotonin widely used to relieve PMS and menstrual pains. Accordingly, the yeast extract containing an excess of tryptophan and niacin is obviously effective in relaxing PMS and menstrual pains. The presence of excess niacin structurally similar to serotonin, which is a kind of neurotransmitter synthesized in the body, indicates high likelihood of converting into serotonin and thus optimizes the autoregulation mechanism in the body to treat pain, chronic fatigue, and depression.

Fourth, thiamin in the yeast extract composition is involved in the nervous system, for example, for normal heart and nerve tissue activities. Low blood thiamin content results in depression, emotional instability, somnipathy, irritability, hyperaction, etc. However, these symptoms are known to disappear by the administration of thiamin (Professor Rus Harrel, Colombia University in New York). Therefore, PMS in women, such as depression, emotional instability, somnipathy, and irritability, can be treated with the thiamin-rich yeast extract.

Fifth, riboflavin in the yeast extraction composition is involved in the synthesis of coenzymes, FMN (flavin mononucleotide) and FAD (flavin adenine dinucleotide). These coenzymes help oxidize nutrients for energy production and synthesize red blood cells and adrenocortical hormone. Accordingly, riboflavin is essential in the period of menstrual bleeding.

Sixth, the mineral selenium in the yeast extract has an antioxidant effect about 1,900 times greater than vitamin E. Neurotransmitters, including serotonin, synthesized in the body may lose their activity by oxidation in the gastrointestical tract or during transmission. Here, the antioxidant selenium aids the neurotransmitters, such as serotonin, to maximize their effects by preventing oxidation of the neurotransmitters.

Finally, an excess of minerals such as calcium and magnesium are required to relieve menstrual pains. Minerals are rapidly absorbed into the body when taken as a food, and their absorption rate is increased with the supply of amino acids. Since the yeast extract are rich in both minerals and amino acids, the minerals can be easily absorbed into the body to effectively manage PMS and menstrual pains. Through the mechanism as described above, the yeast extract can effectively alleviate PMS and menstrual pains.

In another aspect of the present invention, there is provided a PMS and menstrual pain relaxant composition further including chitosan in addition to the yeast extract.

Chitosan is known as an active material to enhance autoimmune response, to aid in the absorption of calcium, and to regulate cholesterol levels. Calcium is an essential mineral for women, especially in the period of menstrual bleeding to relieve PMS. Chitosan is highly effective in removing foreign substances as well as helps calcium absorption, and thus it is useful in relieving PMS and menstrual pains. According to the present invention, it is preferable to use a water-soluble polymeric chitosan of a molecular weight greater than or equal to about 300,000.

Alternatively, the PMS and menstrual pain relaxant composition according to the present invention may further include a herb, such as Korean angelica root, Salviae Radix, Curcuma aromatica, Zedoariae Rhizoma, mint, liquorice, ginger, Gastrodia, white Atractylis, Cnidium officinale, cinnamon, and ginseng.

The above-listed herbs, such as Korean angelica root, Salviae Radix, Curcuma aromatica, liquorice, mint, ginseng, cinnamon, and ginger, are known to aid in the circulation of blood, the dispersal of extravasated blood, the alleviation of pain, the increase of appetite, the activation of metabolism, especially for woman (refer to a text of herbalogy). Congestion due to the non-smooth circulation of blood during the menstrual cycle may cause menstrual pains. The above-listed herbs can alleviate the menstrual pains.

Gastrodia, cnidium officinale, white Atractylis, Zedoariae Rhizoma, etc. aids in the brain blood circulation and in the generation of neurotransmitters to clear head.

Each of the components of the PMD and menstrual pain relaxant composition is ground and mixed with the composition of about 10-90% by weight yeast extract, about 50-80% by weight chitosan, and about 5-80% by weight the herb, based on the total weight of the composition. If excess yeast extract is added, a feeling of langor may result for a patient with mild pain, but an effective reduction in pain results for a patient with severe pain. Therefore, it is preferable to adjust the amount of yeast extract added within the above range.

If excess chitosan is added, the acidity of chitosan itself may act as a stimulus in the body to cause excess tension or stress in a patient with severe menstrual pains, but no adverse effect on a normal person.

The lower limits of the yeast extract and chitosan added are determined to be at least about 10% by weight and about 5% by weight, respectively, to appropriately induce the relaxation effect of the yeast extract and the stimulation effect of the chitosan for effective metabolism in the body. The upper limits of the yeast extract and chitosan added are determined to be about 90% by weight and about 80% by weight, respectively, to appropriately induce the relaxation effect of the yeast extract and the stimulation effect of the chitosan for effective metabolism in the body.

The lower limit of the herb is determined to be at least about 5% by weight by considering its effect of dispersing the extravasated blood remaining after the menstrual cycle. An effective PMS and menstrual pain relaxant composition according to the present invention can be prepared with the addition of at least about 5% by weight the herb.

A method for preparing the yeast extract according to the present invention will be described in greater detail.

In the preparation of the yeast extract according to the present invention, yeast is additionally subjected to a stress, such as high-temperature heating, ultrasonic waves, and vibrations, to secrete stress-resistant substances. This additional step performed at a temperature higher than a lethal temperature of the yeast induces autolysis to utilize active components of the yeast biomass. Here, the yeast's ability to selectively permeate the cell wall is lost, and the cell wall is destroyed by enzymes existing in the yeast, such as protease, lipase, invertase, maltase, zymase, etc. Through the autolysis, a variety of taste components, including amino acids, such as glutamic acid, and nucleic acid metabolites, such as 5'-AMP, are released (Hyung-ik Song and Jung-yeub Shin, Contemporary Fermentation Engineering, 1998, 5:189). Simultaneously, pappain as a protease is added to efficiently give activities as an anti-stress agent, an anti-fatigue agent, and a neurotrophic factor.

Temperature is one of important factors affecting the growth and survival of yeast. Most microorganisms are mesophilic. Yeast has a limited growth temperature of 20-46.degree. C. Saccharomyces cerevisiae, mesophilic yeast, is subjected to a mild thermal shock at 37.degree. C. to induce cell resistance to a lethal temperature of 48-55.degree. C. The resulting thermally resistant cells produce the diose trehalose and effective thermal shock proteins of 90 kDa, 70 kDa, and 60 kDa even with a rise of only 5.degree. C. in temperature from their optimal growth temperature of 35.degree. C. (Michell L. Deegenaars and Kenneth Watson, Environmental Microbiology, the edition of August, 1998). This report supports the fact that yeast produces anti-stress proteins when subjected to thermal stress. Based on this fact, the inventor has additionally induced the synthesis of anti-stress substances by heating at high-temperature and by applying ultrasonic waves and vibrations in the preparations of the yeast extract and yeast-derived peptide according to the present invention. In addition, the method for preparing the yeast extract and yeast-derived peptide according to the present invention involves hydrolying the yeast protein with an addition of a protease while autolysing the yeast at a high-temperature of 35-70.degree. C., and centrifuging the hydrolytes to separate a supernatant.

To investigate the anti-stress capability of the yeast extract and yeast-derived peptide according to the present invention as a neurotrophic factor, an immobile stress test was performed based on the Brekhman and Dardymov method. In general, a series of bioreactions occurring due to stress are initiated by the central nervous system's detection of an external stimulus. Adrenocortical hormone secretion is affected according to the external stimulus to cause changes in the weight of liver, thymus, thyroid gland, and spleen and a reduction in the number of immunocytes. Also, lactate dehydrogenase (LDH) and alkaline phosphatate (ALP) levels in blood are known to change due to an effect of corticosteroid. Changes the above-listed indices have been applied to evaluate anti-stress capability (Cristina, J., Hans, W. and Hans, M., Haematological changes during acute mental stress, Bri. J. Heanat 71:564, 1971; Conner, R. L., Vernikos-Danelis, J. and Levine, S., Stress, fighting and neuroendocrine function, Nature, 2:564, 1971; Munck, A., Guyre, P. M. and Holbrook, N. J., Physiological functions of gluococorticoids in stress and their relation to pharmacological actions, Endocrine Review, 6:25, 1984). As a result of the immobile stress test with rats based on the above anti-stress indices, where changes in the weight of organs and in the level of hematobiochemical factors were measured, the yeast extract and yeast-derived peptide according to the present invention was verified to have the anti-stress effect.

A swimming endurance test was performed to measure the anti-fatigue capability, a kind of anti-stress test, of the yeast extract and yeast-derived peptide according to the present invention. In general, stress is known to affect bipolar affective reaction, motional function, and autonomous function (Yei-wan Hwang, Psychosomatosis, pp. 17-28, 33-49, and 272, Haenglim Publishing Co., Seoul, Korea). Excess exercise, such as swimming, is considered as a stress. Therefore, the swimming endurance test performed using an experimental swimming pool, where excess swimming was forced to measure the powder of endurance, to determine an anti-fatigue effect is considered to be suitable for measuring an anti-stress effect. As a result of the test, the yeast extract and yeast-derived peptide according to the present invention was proven to have excellent anti-stress and anti-fatigue effects.

As another test to measure the anti-stress effect of the yeast extract and yeast-derived peptide according to the present invention, the effect of regulating the autonomic nervous system was determined with patients with anxiety. 200 g of the yeast hydrolyte was orally administered three times a day for 1 week, and the effect of regulating the autonomic nervous system was measured using a stress measurement device (SA-2000, Medicore, Korea). The cardiac cycle (heart rate variability) obtained through power spectral density (PDS) analysis was analyzed for each 5-minute segment to obtain a 5-minute total power. As a result of the autonomic nerve regulation test with the administration of the yeast extract of the invention, autonomic nerve regulation and the ability to regulate against stress- and pain-inducing substances were enhanced with the expectation of its effect on painful PMS similar to stress symptoms.

To verify the bioactivity of the mineral-enriched yeast extract according to the present invention greater than other peptide substances, a macrophage activity test was performed on the mineral-enriched yeast extract to measure immunity. Macrophages, immunocytes involved in both congenital and adaptive immune systems, are critical in the cell-mediated immunity (CMI) to provide antigens for inducing lymphocytes through digestion and successive decomposition and other processes of externally introduced substances, to secrete specific substances, such as immunoregulatory cytokine, and to produce nitric oxide (NO) having the function of killing foreign invaders, antigens. Macrophages are divided into inflammatory macrophages and activated macrophages depending on activity. Inflammatory macrophages are produced when exposed to inflammation inducer substances, such as thioglycolate. The inflammatory macrophages have phagocytic activity and surface adherence, and increase the secretion of prostaglandin, the ability to synthesize protein of a variety of enzymes, such as plasminogen activating enzyme, elastase, collagenase, etc., cell size, and the release of a number of cellular discharges.

Macrophages activated by cytokines, such as IFN-.gamma. and TNF, and lipopolysaccaride (LPS) from gram-negative microorganisms have anti-cancer and anti-microbial effects. In the cytotoxic mechanism of the macrophages with respect to cancer, cytokines, such as TNF-.alpha., IL-1, IL-6, IL-8, and IL-12, hydrogen peroxide (H.sub.2O.sub.2), nitric acid (NO), and cytolytic protease which are released from the activated macrophages are known to have a toxicity with respect to cancer cells. The macrophages activated by such lectins, large molecular proteoglycan, and polysaccharide primarily suppress oncogenesis and then tumor metastasis, and have the ability to distinguish oncocytes and normal cells. Although a target structure the macrophage can detect has not been identified yet, unlike the phagocytosis mechanism of the macrophage, the macrophage adheres to a target oncocyte to release lysosomal enzymes before lysis of the target oncocyte. Meanwhile, non-activated macrophages have a weak toxicity to tumor cells and thus needs to be activated for enhanced oncolytic abilities. This fact supports that immunotherapy to lead macrophage activation can be an effective therapy. Accordingly, the microphage activity was tested for the mineral-enriched yeast extract according to the present invention. As a result, it was proven that the mineral-enriched yeast extract according to the present invention is effective in enhancing immunity.

The bioactivity of the mineral-enriched yeast extract according to the present invention greater than other peptide substances was additionally verified through an intestinal immunity activity test to measure immunity. Immunocytes constitute tissues or organs, called the lymphatic system, for effective immune reactions. The lymphatic system is classified into primary (or central) lymphoid organs, including bone marrow and thymus, for producing lymphocytes, and secondary lymphoid organs, including lymph nodes, spleen, and mucosa-associated lymphoid tissues (MALT), for providing conditions or environments for the contact of the lymphocytes and antigens and the interactions between lymphocytes. The MALT, which is located at a region susceptible to microorganisms, plays an important role in regional, mucosal immune reactions, so it is called the mucosal immune system. The MALT, which induces a defective reaction against antigens entering the body via ingestion or inspiration, is classified into the gut-associated lymphoid tissues (GALT) located in digestive track, the bronchous-associated lymphoid tissues (BALT) located in the musculi canal, and the nasal-associated lymphoid tissues (NALT) located in the junction of the plate and nose. Among these types of lymphoid tissues, the GALT as the largest lymphoid tissue present in the intestinal mucosa is especially significant in the body protective system. The alimentary mucosa having a large surface area is always exposed to a number of different microorganisms and plant-derived heterologous proteins or compounds, which supports the immunological significance of the GALT.

Peyer's patches used in the examples according to the present invention to be described later, which have the typical structure of the GALT, are an aggregate of nodi lymphatici mesenterici distributed in the small intestinal mucosa. Peyer's patches are easily observed, and most are observed in the ileum. Activated lymphocytes in the Peyer's patch release a variety of cytokines, including IL-6 and GM-CSF (granulocyte macrophage-colony stimulating factor), to regulate immune or inflammatory reactions by regulating the growth, migration, proliferation of bone marrow cells, white blood cells, and hematogenic cells. Based on this fact, in an example according to the present invention to be described later the degree of proliferation of bone marrow cells by cytokines secreted with the activation of the Peyer's patch cells using the mineral-enriched yeast extract according to the present invention was measured to determine enhanced immunoactivity.

In general, a stimulus to the human body is known to be injurious if it's intensity is over the limit, thereby causing tension headaches, migraine headaches, hypertension, indigestion, fatigue, or generalized headaches. If such a stimulus is prolonged chronically, non-specific general adaptative syndromes, such as neuropathies or gastropathies, may result (Yei-wan Hwang, Psychosomatosis, pp. 17-28, 33-49, and 272, Haenglim Publishing Co., Seoul). As described above, the yeast extract and yeast-derived peptide according to the present invention have an anti-stress effect and can be used as a tranquilizer or a relaxtant. In particular, it is greatly expected to use the yeast extract and yeast-derived peptide according to the present invention as a sleeping aid for nervous people with sleeping problems. The yeast extract and yeast-derived peptide according to the present invention are useful as a source for auxiliary health foods and special nutritional foods having an activity as a brain-neurotrophic factor for relieving the above symptoms.

As described above, the yeast extract and yeast-derived peptide according to the present invention are effective in the regulation of the autonomic nervous system, for example, in alleviating stress and inducing deep sleep. The yeast extract and yeast-derived peptide according to the present invention are believed to be an effective neurotrophin for normalizing the neurological function without any side effect for excess doses.

Unlike conventional side-effect inducing therapeutic drugs for nervous and stress symptoms, such as a tranquilizer, an anti-anxiety agent, a sleeping drug, etc., the yeast extract and yeast-derived peptide according to the present invention prepared from a natural source and having anti-stress and neurotrophic effects can effectively regulate the autonomic nervous system to alleviate a number of stress symptoms without any side effect. In particular, the yeast extract and yeast-derived peptide according to the present invention are applicable as a tranquilizer, an anti-stress agent, a sleeping aid, an anti-fatigue agent, and PMS and menstrual pain relaxants.

The yeast extract and yeast-derived peptide according to the present invention having the anti-stress and neurotrophic effects are available as a substitute for conventional side-effect inducing psychotrophic drugs in the preparation of medicines, active foods, medicines and feed for animals, etc. For use of the yeast extract and yeast-derived peptide according to the present invention as therapeutic agents, a variety of known pharmaceutical methods can be applied in the preparation of those drugs. The yeast extract and yeast-derive peptide according to the present invention may be processed alone or mixed with a pharmaceutically safe carrier, vehicle, diluent, etc. into powder, granule, tablet, capsule, or injection form to be orally or non-orally administered.

When the yeast extract and yeast-derived peptide according to the present invention are used as a therapeutic agent, its dose can be appropriately determined depending on the age, sex, state, and symptom of a patient. Preferably, the yeast extract or yeast-derived peptide according to the present invention is used at a dose of about 10-500 mg a day for adults. For a patient with insomnia, a capsule containing 250 mg of the yeast extract or yeast-derived peptide powder is administered twice a day.

For use of the yeast extract according to the present invention as a PMS and menstrual pain relaxant, the yeast extract is ground, and capsules are filled with the yeast extract powder. Alternatively, the yeast extract powder may be processed into tablets. When a dose of 1-2 capsules (or tablets) two or three times a day, each capsule (or tablet) containing 180 mg of the yeast extract powder, is taken with excess water, PMS, menstrual pains, hysterorrhea, and dysmenorrhea in females are effectively alleviated.

The yeast extract and yeast-derived peptide according to the present invention or a composition containing the same are applicable in the preparation of heath beverages effective in relieving females of PMS, menstrual pains, hysterorrhea, etc.
 

Claim 1 of 6 Claims

1. A method for treating a disorder comprising administering to an animal having a disorder an amount of a yeast extract effective in treating the disorder, wherein the disorder is clinical depression, anxiety, stress, or fatigue, wherein the disorder is caused by excessive reuptake of serotonin and norepinephrine.

 

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