KR101482044B1 - Food composition comprising fermented product of Pleuropterus multiflorus and ginger for alleviating or preventing degenerative brain disorder and method of producing the same - Google Patents

Abstract

Disclosed are a composition and a food composition containing a fermented product of Pleuropterus multiflorus and ginger for preventing or treating degenerative brain disorder and a method for preparing the same. The fermented product of the present application is enriched with minerals and amino acids, and can be useful for the treatment or prevention of regenerative brain disorder by inhibiting brain cell damage and inflammation through the activation of nitrogen monoxide SOD and the inhibition of NO generation.

Description

[0001] The present invention relates to a food composition for preventing or ameliorating degenerative brain diseases, which comprises fermented fruits and vegetables,

Field of the Invention The present invention relates to a field of functional natural products, and more specifically to a fermented product of sewage and ginger.

As an aging society, the incidence of chronic degenerative diseases has been steadily increasing over acute communicable diseases, and the increase of brain diseases among chronic degenerative diseases has become a serious social problem. However, to date, effective drugs and therapies for the prevention and treatment of degenerative brain diseases have not been developed.

It is known that degenerative brain diseases are related to various degenerative brain diseases such as inflammatory mediators of microglia, which are immune cells in the brain (Han, IO, et al., Neurosci Lett.325: 9-12, 2002) .

Currently, various methods have been provided to treat degenerative brain diseases, but there are drawbacks such as adverse effects and dosage methods which are indicative of hepatotoxicity. In order to overcome these problems, studies on various natural extracts have been conducted recently, and the demand for functional foods using natural extracts is steadily increasing.

Korean Patent Laid-Open Publication No. 2005-0008598 relates to a composition containing a bellflower extract, which discloses a pharmaceutical composition for preventing or treating degenerative brain diseases or enhancing memory ability comprising bellflower extract as an active ingredient.

Korean Patent Laid-Open Publication No. 2005-0101818 relates to a herbal composition, which discloses a composition for the prevention and treatment of degenerative brain diseases that exhibits a neuroprotective effect including hwanggi, hanchu, turmeric and gugija.

Therefore, it is inevitable to develop materials for prevention or treatment of degenerative brain diseases, which exhibit safer and superior effects derived from natural substances. The market demand for development of excellent materials having a new action mechanism from various medicinal plants and agricultural life resources Is very high.

The present invention provides an extract, a composition and a health functional food which can be effectively used for the treatment or prevention of degenerative brain diseases using natural substances.

In one embodiment, the present invention provides a pharmaceutical composition for the treatment or prevention of degenerative brain diseases comprising a fermented product of desserts and ginger as an active ingredient, or a food composition for improving, alleviating and / or preventing degenerative brain diseases.

The fermentation according to the present invention can be used with various strains as long as they exhibit the effect according to the present invention. In one embodiment lactic acid bacteria such as Streptococcus sp. , Pediococcus sp. sp . ), Leuconostoc ( Leuconostoc sp . ), The genus Lactobacillus sp . ) Or Bifidobacterium sp . ). ≪ / RTI >

The present sewage and ginger fermented product and a composition containing the same promote the activity of superoxide dismutase (SOD), which is capable of removing superoxide radical anions, and inhibit the secretion of nitrogen monoxide from microgranules to prevent or treat degenerative brain diseases . Thus, the composition according to the present invention is useful for the treatment of degenerative brain diseases caused by the production of nitrogen monoxide and superoxide radicals, for example degenerative brain diseases such as Alzheimer's disease, Parkinson's disease, meningitis, stroke, dementia, Huntington's disease or degenerative brain diseases such as Creutzfeldt- Can be usefully used.

The compositions according to the present invention were also fortified with antioxidant components and minerals by fermentation.

In the composition according to the invention, the gasses and ginger may be contained in a weight ratio of 1: 0.1 to 1: 1, in particular 10: 4.

The composition according to the present invention can be provided not only as a pharmaceutical composition but also as a food composition for prevention or remedy of degenerative brain diseases including a dessert and a ginger fermented product. The food composition can be used as a health supplement, a functional food, Lt; / RTI >

In another aspect, the disclosure also provides a method of making a sewage and ginger fermentation product. In one embodiment, the method comprises mixing the gut and ginger and adding sugar to the mixture (or mixing the gut, ginger and sugar); And culturing the mixture after inoculating the lactic acid bacteria.

The process according to the invention can be carried out in particular in a stationary state without stirring. Also, in the method of the present invention, the sewage and ginger are contained in a weight ratio of about 1: 0.1 to 1: 1, and in particular, the above-mentioned sewage, ginger and sugar are contained at a weight ratio of 10: 4: 2. Sauce and ginger are not only dried, but also raw, powdered, and sliced.

After the lactic acid bacteria inoculation, the culture is incubated at about 30 ° C to 40 ° C for about 5 to 16 days, particularly at 38 ° C for 10 days.

The lactic acid bacteria used in the method of the present invention can be referred to the above.

The composition comprising the fermented product of the present invention and the health food containing the fermented product of the present invention is not only strengthened with minerals and amino acids due to fermentation but also inhibits damage and inflammatory reaction of brain cells through activation of NO and SOD activation Can be usefully used for preventing, treating, ameliorating and / or alleviating degenerative brain diseases.

FIG. 1 is a graph showing the survival rate of BV-2 cells according to one embodiment of the present invention. FIG.
FIG. 2 is a graph showing the activity of inhibiting nitrogen monoxide formation according to the concentration of the fermented product of sodium chloride and ginger according to an embodiment of the present invention.
FIG. 3 is a graph comparing the activity of inhibiting nitrogen monoxide formation of the fermented product of the gut and the ginger according to an embodiment of the present invention, in comparison with the control group.

In one aspect, the present invention relates to a composition for treating or preventing a degenerative brain disease comprising an active ingredient of a fermented product of gut and ginger.

Pleuropterus multiflorus TURCZ. Is a perennial vine plant of the genus Asclepiadaceae, which is called white cucumber, and has a scientific name of Cynanchum wilfordii Hemsley. It is also known as other names such as mockery, leopard grass, sunshine, new mockery, multipurpose roots, and is one of the herbal medicines prescribed for prevention of hair loss in one room, prevention of arthritis and nourishment. The major components include polyoxypregnane glycoside, aglycone, and gagaminine (DU Lee, et al., Planta Medica., 62, 485-487 , 1996, T. Nomura and H. Mitsuhashi, Pharm. Bull., 20, 2065-2067, 1972).

As used herein, roots are mainly used for roots, but seed roots, leaves, roots, roots and seeds may also be used as long as they include physiological activity.

Ginger (Zingiber officinale Roscoe) is a perennial plant belonging to the genus Zingiberaceae. Its rootstock grows sideways and is fleshy, lumpy, yellowish, with a pungent and fragrant smell. In the oriental medicine, dried roots are used as medicines, which are used to treat chills, fever, headache and sputum caused by cold. Pharmacological actions promote gastric juice secretion, digestive power enhancement, cardiac excitement and blood circulation promotion. Jitoe et al., Tetrahedron Lett., 35, pp 981-984, 1994) and anti-inflammatory effects (Ozaki et al. Chem. Pharm. Bull., 39, pp2353-2356, 1991; Jeenapongsa et al., J. Endopharmacol., 87, pp. 143-148, 2003). The main ingredients are gingerone, shoagaol, tetralin, zingiberene, curcuminoid, flavonoid glycoside, sesquiterpenoid, ) And polyphenol (polyphenol).

As used herein, roots are mainly used for roots, but seed roots, leaves, roots, roots and seeds may also be used as long as they include physiological activity.

The term "fermented product" as used herein refers to a final product obtained through a fermentation process according to the present invention, and it is possible to remove the solid content after fermentation. The material used can be removed, and this is included herein. In one embodiment, the solid content after fermentation is sieved to a sieve, such as 100 mesh. Various fermented lactic acid bacteria that produce lactic acid may be used for the fermentation of the present invention.

Fermentation can be carried out using a variety of fermenting bacteria such as Saccharomyces cerevisiae , Bacillus subtilis , Acetobacter aceti and / or lactic acid bacteria, In embodiments, lactic acid bacteria are used.

The lactic acid bacteria may be classified according to characteristics such as the bacterial shape, fermentation method, aerobic / anaerobic characteristics, and lactic acid bacteria belonging to various categories may be used to achieve the effect of the present invention. For example, Streptococcus sp ., Pediococcus sp ., Leuconostoc sp ., Lactobacillus sp ., Or Bifidobacterium sp . ), But is not limited thereto. In one embodiment, Lactobacillus sp ., For example, Lactobacillus acidophilus may be used, but is not limited thereto.

The sewage and ginger used in the fermentation composition according to the present invention may be included in a weight ratio of about 1: 0.1 to 1: 1, and in one embodiment, in a weight ratio of 1: 0.4.

The ginger and the sewage according to the present invention may be fermented and mixed, or mixed in the weight ratio as described above and then fermented, respectively, so long as they satisfy the weight ratio as described above.

Sodium and ginger used in the present invention are produced by fermenting the active ingredients in the raw material to various oil soluble components, and in particular, various enzyme components are produced through fermentation, low molecular weight proteins are produced, amino acids are strengthened, The components were transferred to the active form and the mineral became enriched.

The degenerative brain disease of the present invention is a brain-induced disease of the degenerative disease caused by age, and the degenerative brain disease in which the composition of the present invention is effective is Alzheimer's disease, Parkinson's disease, meningitis, stroke, dementia, Huntington's disease, - Jakob disease, and the like.

Degenerative brain diseases are caused by the fact that the specific brain cell group of the brain and spinal cord gradually lose its function and the number of brain cells decreases due to the unknown cause to date, and the damage and death of brain cells, the formation of synapses or functional problems, ≪ / RTI > The brain cells are damaged by free radicals, which are hydroxyl radicals, superoxide radicals and nitric oxide (NO).

It is known that the inflammatory response caused by the activation of macrophage cells, a macrophage of the nervous system, induces secondary neuronal apoptosis and is involved in the induction of chronic neural degeneration. In other words, it produces and secretes cytotoxic cytotoxic substances such as TNF-α, interleukin-1 (IL-1β) and interleukin-6 (IL-6) , Intracellular signal transduction, etc. However, when the nitrogen monoxide is excessively produced, an increase in peroxynitrite, an increase in superoxide by NADPH oxidase, and an increase in cyclooxygenase are observed. And increases oxidative stress by inducing oxygen species increase by activity increase.

Long-term neuronal degeneration may persist if the abnormal microglial response is maintained chronically. In particular, nitrogen monoxide is a small, unstable, and toxic free free radical, which ultimately damages surrounding neurons as well as causing degenerative diseases such as inflammation, meningitis, Alzheimer's disease, and Parkinson's disease (Kasckow et al., 2003; Ward et al., 2001; Lezoualch et al., 2000).

The superoxide radical is a highly toxic radical generated in the enzymatic and non-enzymatic stages of aerobic cells and is produced as a result of various oxidoreductases in vivo. It is highly reactive and is involved in the initiation phase of the oxidation reaction related to aging I am involved. In addition, it has been known to induce oxidative damage to lipids, proteins, DNA, etc. in the production of other reactive oxygen species such as hydrogen peroxide (R. Ishida et al., Biochem. And Biophy. Research Comm. , 1432-1438, 1975).

Therefore, it is possible to inhibit or treat the onset of degenerative brain diseases by inhibiting the production of nitrogen monoxide and superoxide radicals.

The present sewage and ginger fermented product and a composition containing the same promote the activity of superoxide dismutase (SOD), which is capable of removing superoxide radical anions, and inhibit the secretion of nitrogen monoxide from microgranules to prevent or treat degenerative brain diseases .

The term "treatment ", as used herein, refers to any action that improves or alters the symptoms of degenerative brain disease or other conditions resulting therefrom upon administration of the extract or composition comprising it. If the person skilled in the art is familiar with the present invention, refer to the data presented by the Korean Medical Association to know the precise criteria of the degenerative brain disease or disease for which the composition of the present invention is effective, and judge the improvement, You can do it.

As used herein, the term "prophylactic " means any act that inhibits or delays the onset of degenerative brain disease or other diseases resulting therefrom upon administration of the extract or composition comprising it. It will be apparent to those skilled in the art that the extract of the present invention which is therapeutically effective for degenerative brain diseases can prevent the early symptoms of degenerative brain diseases or such diseases when taken before they appear.

The pharmaceutical composition comprising the present hosiery and ginger fermented product may be administered simultaneously or sequentially, and these compounds may be administered alone or in combination with other pharmaceutical active ingredients for the treatment of symptoms of degenerative brain diseases such as Alzheimer's disease .

The compositions comprising the present sewage and ginger fermentations may be formulated with suitable excipients such as suitable carriers, diluents, preservatives, stabilizers, wetting agents, emulsifiers, solubilizers, sweeteners, colorants, . Specific examples include lactose, dextrose, sucrose, sorbitol, mannitol xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone Water, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium, stearate, mineral oil and the like.

The method for administering the pharmaceutical composition for prevention or treatment of degenerative brain diseases including the sewage and ginger fermented product of the present invention can be easily selected according to the formulations and can be administered to mammals such as livestock and human by various routes. For example, it may be formulated in the form of powders, tablets, pills, granules, dragees, hard or soft capsules, liquids, emulsions, suspensions, syrups, elixirs, external preparations, suppositories, sterilized injection solutions, Orally or parenterally. In particular, oral administration is preferred.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid formulations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used diluents, various excipients such as wetting agents, sweeteners, have.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

Further, pharmaceutical compositions comprising the present Sauce and Ginger fermentations may be prepared using any suitable method known in the art or as described in Remington ' s Pharmaceutical Sciences (recent edition), Mack Publishing Company, Easton PA Method. ≪ / RTI >

The dosage of the pharmaceutical composition containing the present sewage and ginger fermented product may vary depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, Effective dosages of the sewage and ginger fermentations are typically about 1 to 100 g / day, especially about 10 to 50 g / day, more preferably about 30 g / day, for an adult (60 kg). It will be apparent to those skilled in the art that doses may be additive or subtracted, as the dosage can vary depending on various conditions, and thus the dose is not intended to limit the scope of the invention in any way.

The number of administrations can be administered once or several times a day within a desired range, and the administration period is not particularly limited. In addition, the composition containing the extract of the present invention may be added to an arbitrary food and be routinely ingested as such, in addition to oral administration.

The pharmaceutical composition comprising the present sewage and ginger fermented product not only provides an excellent therapeutic effect against degenerative brain diseases, but also has no toxicity and side effects caused by drugs, and can be used safely even when taken for a long time. Accordingly, the sewage and ginger fermented product of the present invention can be used as a main ingredient, a food ingredient, and a food additive.

Therefore, from this point of view, the present invention can be provided as a food, for example, a health supplement, a functional food, a food additive or the like, the composition comprising the above-described sewage and ginger fermented product of the present invention.

As used herein, the term "food" means a natural or processed product containing one or more nutrients, preferably a state of being ready to be eaten through a certain degree of processing, It is intended to include food, food additives, functional foods and beverages as a meaning.

As used herein, the term "functional food" refers to a food group that has been imparted with added value to function or express the function of the food by physical, biochemical, or biotechnological techniques, , And the body control function of disease prevention and recovery is designed to be fully expressed in the living body, and in particular, includes "health functional food".

As used herein, the term "health supplement food" is a food prepared by using raw materials or ingredients having useful functions in the human body, and is useful for health and other purposes such as controlling nutrients and physiological actions on the structure and function of the human body For the purpose of health assisting to obtain the effect, it refers to a food which is made by processing a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. ingredients contained in a food raw material.

Particularly, the functional food or the health supplement food according to the present invention means a food having an effect of improving symptoms of degenerative brain disease, or alleviating or alleviating the symptoms of degenerative brain disease, compared with before the food is consumed. If you are familiar with the technology, you can refer to the data provided by the Korean Medical Association to know the precise standards of these diseases and to judge the degree of improvement.

The functional food and the health supplement food may further include a food-acceptable food supplementary additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of a functional food.

Foods to which the present sewage and ginger fermentation products can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and functional foods. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods But are not limited to, fruits, vegetables, beverages, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.). The food, beverage or food additive may be prepared by a conventional production method.

Drinks in the present application mean a general term for drinking or enjoying a taste, and are intended to include functional beverages. The beverage is not particularly limited as long as it contains the above-mentioned sucrose and ginger fermented product of the present invention as an active ingredient as an essential ingredient at the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient ≪ / RTI > Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and Xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention. May be further contained.

In the present invention, the functional beverage is used to control the bio-defense rhythm of the beverage group or beverage composition to which the added value is imparted so that the function of the beverage functions for a specific purpose using physical, biochemical, biotechnological techniques, Means a drink which is processed by designing the body so as to sufficiently express the body control function with respect to the living body, and preferably means a drink for the improvement of degenerative brain diseases.

The functional beverage is not particularly limited to the other components other than those containing the present sewage and ginger fermented product as essential components in the indicated ratios and may contain various flavors or natural carbohydrates as additional components such as ordinary beverages . Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and xylitol , Sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not so critical, but may be selected in the range of 0.001 to 20 parts by weight per 100 parts by weight of the fermentation product of the present invention.

In the food for the purpose of improving degenerative brain diseases, the present sewage and ginger-extracted fermented product comprises about 0.01 to 90% by weight, about 0.1 to 80% by weight, about 1 to 70%, about 1 % To 50%, about 1% to 40%, about 1% to 30%, about 1% to 20%, but does not exclude the range. For example, the beverage composition may comprise 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 mL.

In another aspect, the present invention is also directed to a method of making a fermentation product of a gut extracted and used in the composition of the present invention.

The method according to the present invention comprises the steps of mixing a mixture of sewage and ginger followed by adding sugar (for example, sugar) to the mixture (or mixing the sewage, ginger and sugar); And inoculating the mixture with lactic acid bacteria, followed by culturing at 30 ° C to 40 ° C for 5 to 16 days.

In one embodiment, the fermentation proceeds without stirring. When lactic acid bacteria are used, the amount of dissolved oxygen in the culture liquid is increased during stirring, and other intermediate metabolites other than lactic acid fermentation are produced in a complex manner. However, when stirring is not performed, lactic acid, which is a pure fermentation product of lactic acid bacteria, is mainly produced and the effect of improving the flavor is excellent.

Sewage and ginger used in the process according to the invention are as mentioned above. In addition, the desserts and ginger may be used in dried form, unprocessed form, or may be pretreated in the same manner as cutting or grinding.

In one embodiment, the ginseng and ginger are used in a weight ratio of about 1: 1 and 1: 1, especially about 1: 0.4. In another embodiment, the safflower, ginger and sugar are used in a weight ratio of 10: 4: 2.

In one embodiment, the incubation is carried out at a temperature and for a time sufficient for the fermentation to produce sufficient amounts of the oil component. Lt; RTI ID = 0.0 > 37 C < / RTI > for about 14 days.

Hereinafter, embodiments are provided to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited to the following examples.

Example 1 Preparation of Fermented Sauce and Ginger 1

10g of dried slices purchased from a herbal medicine wholesaler in Yeongju city, Gyeongbuk province was mixed with 100ml of distilled water and added with 0g, 2g, 4g, 6g, 8g, and 10g of dry ginger slice raw materials, respectively, based on the sterilized liquid medium. Five ml of Lactobacillus sp. Culture broth (total 2x10 ⁷ CFU) isolated from kimchi was added to one of six kinds of medium, followed by static culture at 37 ° C for 14 days without stirring at 37 ° C.

Example 2 Preparation of Fermented Sauce and Ginger 2

1 g, 2 g, 3 g, 4 g, and 5 g of sugar as sugar components were added to the basic medium containing 10 g of sucrose, 4 g of ginger, and 100 ml of distilled water in order to increase the fermentation efficiency including the shortening of fermentation time of lactic acid bacteria and the increase of lactic acid bacteria. After sterilization, 5 ml of the lactic acid bacterium solution of Example 1 was inoculated in each case, followed by stationary culture at 37 캜 for 14 days without stirring at 37 캜.

Example 3 Analysis of viable cell count of fermentation broth

The viable cell counts of the hydrogen sulfide and ginger fermentation broth prepared in Example 1 were analyzed by measuring the absorbance (600 nm). The results are shown in Table 1. As a result, the highest absorbance value was obtained in a medium containing 10 g of Sasa and 4 g of ginger. Then, the sample of the culture broth was diluted with MRS medium (1 L of component and content of Pancreatic digest of gelatin 10.0 g, Beef extract 8.0 g, Dextrose 20.0 g, Dipotassium phosphate 2.0 g Polysorbate 80 1.0 g Sodium acetate 5.0 g Ammonium 2.0 g of citrate, 0.2 g of magnesium sulfate and 0.05 g of manganese sulfate), and the number of viable cells was measured. As a result, the highest viable cell count was 2.0 × 10 ⁵ CFU / ml in a medium containing 10 g of bagasse and 4 g of ginger, The mixing ratio composition was found to be most effective for the growth of lactic acid bacteria.

White ginseng (g): ginger (g) Absorbance (A _600nm ) Number of living cells (CFU) / ml 10: 0 0.12 0.7 x 10 ⁴ 10: 2 0.78 1.2x10 ⁵ 10: 4 0.83 2.0x10 ⁵ 10: 6 0.71 1.5x10 ⁵ 10: 8 0.67 1.1x10 ⁵ 10: 10 0.54 0.9x10 ⁴

Example 4 Analysis of components and efficacy of fermentation broth 1

The number of viable cells contained in the culture broth was checked by the smear method in the MRS medium at intervals of one day from the fifth day after the fermentation of the fermented broth and ginger according to Example 2. The culture time, Antioxidant enzyme activity and total polyphenol content are shown in Table 2.

The medium supplemented with 2 g of sugar showed the highest viable cell count of 0.8 × 10 ⁷ CFU after 10 days of culture, and it was judged to be the most suitable medium for the growth of lactic acid bacteria.

The activity of SOD (Superoxide dismutase), a typical antioxidant enzyme, was measured by SOD assay kit (Sigma Aldrich, USA) according to the manufacturer's method. In summary, the excess oxide radical anion reacts with nitroblu tetrazolium (NBT) to form a blue color. If an antioxidant is present in the sample, the blue oxide is discolored by interfering with the formation of the excess oxide radical anion -NBT complex. After the reaction, the absorbance at 560 nm was measured using a spectrophotometer, and the SOD activity was calculated as follows.

         SOD (%) = (1- (A-B) / C) 100

A: Absorbance is measured after addition of sample to xanthine oxidase reagent

B: Absorbance measurement after addition of blank buffer instead of xanthine oxidase reagent

C: Absorbance measurement after addition of distilled water to xanthine oxidase reagent

As a result, SOD activity and total polyphenol contents were 43.6% and 65.9 mg / 100g in the case of the medium containing the highest amount of live bacteria in the content of bagasse: ginger: sugar = 10: 4: 2 (w: w: , And it was determined to be a medium composition suitable for improving not only the growth of the cells but also the antioxidation function.

Total polyphenol content was determined by Folin-Ciocalteu's phenol method. To summarize, 1.9 ml of distilled water and 0.2 ml of Folin-Ciocalteu's phenol reagent were added to 0.1 ml of the fermentation broth and reacted at room temperature for 3 minutes. 0.4 ml of a saturated Na ₂ CO ₃ solution and 1.9 ml of distilled water were added and mixed, and the absorbance was measured at 725 nm using a spectrophotometer. The total polyphenol content of the sample solution was calculated as the amount of galactic acid (mg) per 100 g of the sample solution after a standard curve was formed using garlic acid (Sigma, USA).

Back Sash: Ginger: Sugar
(w: w: w) culture
time
(day) The number of live bacteria
(CFU) / ml SOD activity
(%) total polyphenol
(mg / 100g) 10: 4: 0 14 0.2x10 ⁶ 27.4 35.2 10: 4: 1 12 0.1 x 10 ⁷ 34.2 37.9 10: 4: 2 10 0.8 x 10 ⁷ 43.6 65.9 10: 4: 3 11 0.7 x 10 ⁷ 40.1 53.7 10: 4: 4 12 0.2x10 ⁷ 39.2 50.4 10: 4: 5 13 0.6x10 ⁶ 28.4 33.3

Example 5: Analysis of components and efficacy of fermentation broth 2

The sludge and ginger fermented product according to the above Example 2 was subjected to cell-free removal and separation and purification using a filter press and a filter paper, followed by lyophilization And stored at 4 ° C and used as a sample for analysis.

The physicochemical properties of Baekhwa-gum ginger complex lactic acid bacteria fermented were determined by measuring the water solubility, pH, thermal stability, and chromaticity using freeze-dried powder. The results are shown in [Table 3]. At this time, the comparative group was composed of the Bacillus subtilis ginger complex hydrothermal extract, Bacillus subtilis hydrothermal extract, and ginger hydrothermal extract, which were prepared without fermenting the same amount of the composite material with lactic acid bacteria. The hot water extraction conditions were as follows: 1: 5 (w / v) ratio of each raw material and distilled water was extracted at 80 ℃ for 36 hours, and then concentrated under reduced pressure and then lyophilized.

Sucrose and ginger extract fermented water showed higher water solubility and thermal stability than the hot water extracts of the comparative group.

division Complex fermentation product Complex hot water extract Sewage
Hot water extract ginger
Hot water extract receptivity +++ ++ ++ ++ pH 5.8 6.4 6.8 6.2 Thermal stability (-10 ° C to 80 ° C) +++ ++ ++ ++ Color Light brown Brown Light brown Brown

Example 6 Analysis of components and efficacy of fermentation broth 3

Ingredients and efficacy analyzes of the fermented product of Sucrose and Ginger (Sauce: Ginger: Sugar = 10: 4: 2 by weight, 10 days culture) according to Example 2 and unfermented raw materials were carried out. The comparative groups were white sacha extract, ginger extract, unfermented white sacha and ginger complex extract (Sasa: ginger: sugar = 10: 4: 2 weight ratio). SOD activity and total polyphenol content of the fermented product and the comparative group were compared and analyzed.

SOD activity was 43.6% higher than that of the control group, especially about 56% higher than that of the non - fermented complex extract. The total polyphenol content was also about 76% higher than that of the non - fermented complex extract, which showed an amount of 65.9 mg / 100 g and was mixed at the same ratio. Based on the high SOD activity and the polyphenol content, the antioxidant - related enhancement pattern was confirmed.

Sample type SOD activity
(%) total polyphenol
(mg / 100g) White Sachao Extract 21.3 26.5 Ginger extract 15.2 18.9 White Seaweed + Ginger Extract 27.9 37.4 White saury + ginger fermentation 43.6 65.9

Example 7 Analysis of components and efficacy of fermentation broth 4

The major minerals component analyzes of the fermented extracts of Sasa and Ginger were compared and the results are shown in Table 5. Contents of phosphorus, magnesium, calcium and zinc were analyzed by using atomic absorption spectrophotometer as a major physiologically active mineral component. The comparative group was hydrothermal extract of ginseng, hot water extract of ginger, and hydrothermal extract of unfermented ginseng and ginger . The major minerals of the fermented seaweed and ginger extracts were found to increase more than the hot water extracts of Sasa and Ginger before fermentation.

division Complex fermentation product Complex hot water extract Bleached water
extract ginger
Hot water extract Phosphorus (mg / 100 g) 12.1 10.4 10.1 8.2 Magnesium (mg / 100 g) 465 319 412 217 Calcium (mg / 100g) 58 43 34 29 Zinc (mg / 100g) 35 26 28 17

Example 8 Evaluation of cytotoxicity of fermented products of Sasao and Ginger

In order to evaluate the cytotoxicity of the complex fermented product, microsomes (BV-2 cells) were purchased from Kyungpook National University College of Medicine, and basic microbial cell culture was performed using DMEM containing 10% FBS and 1% penicillin / strepromycin , USA) at 37 ° C in a 5% CO ₂ incubator.

Cytotoxicity was measured by the manufacturer's method with CellTiter 96 ^ⓡ AQueous One Solution Cell Proliferation Assay Kit (Promega, USA).

Briefly, the assay was performed to quantitate cell proliferative capacity and cell viability. MTT (3- [4,5-dimethylthiazol-2-yl] -2,5 diphenyl tetrazolium bromide) was transformed into mitochondrial dehydrogenase The amount of coloring matter called formazan is measured. It is effective only for living cells, and the intensity of the color is linear with the number of cells. BV-2 cells were suspended in a medium containing 10% FBS at a concentration of 1.5 x 10 ⁴ cells / well, 200 μl of each was inoculated into each well of a 96-well plate, and cultured for 24 hours. Were cultured for 24 hours. Then, 20 μl of MTS solution was added to the medium without 100 μl of FBS per well, and the reaction was allowed to proceed for 1 hour. Then, the color intensity was measured using a medical enzyme analyzer (Labsystems, FI / Multiskan MS).

The results are shown in FIG. 1, and the fermented product of B. subtilis ginger combined lactic acid bacteria showed no cytotoxicity at a concentration of 500 μg / ml or less, and the same toxicity was not observed even when 0.5 μg / ml of LPS was treated at the same time Respectively.

Example 9 Inhibitory Effect on the Formation of Nitric Oxide (NO) in Fermented Sauce and Ginger

NO concentration was measured by a colorimetric assay kit (Promega, USA) using Griess reagent according to the manufacturer's method. Briefly, BV-2 cells were plated at 1 × 10 ⁵ cells / ml on a 96-well plate, and LPS (Lipopolysaccharide) and the fermented product of the present invention were added to cultured cells as the inflammatory reaction inducer After incubation for 24 hours, 100 μl of each culture was taken from each well. The same amount of Griess reagent (0.1% naphthyl ethylene diamine and 1% sufanilamide in 5% H ₃ PO ₄ ) was added and the mixture was allowed to stand at room temperature for 10 minutes. The total nitrite formation was measured at 540 nm using sodium nitrite (NaNO ₂ ) as the standard curve.

As a result of measuring the NO production amount of the fermented product of Lactobacillus acidophilus Lactobacillus at a concentration range that does not exhibit cytotoxicity, 63.8 μM was produced when LPS inducing inflammatory response to BV-2 cells alone as shown in FIG. It was confirmed that the amount of nitrogen monoxide was decreased with increasing concentration of fermented product at 57.9 μM when treated with 100 μg / ml of lactic acid bacteria, 47.2 μM when treated with 300 μg / ml, and 40.6 μM when treated with 500 μg / ml .

3, the concentration of NO produced after LPS treatment in BV-2 cells of the hot water extract, hot water extract of ginger, and hydrothermal extract of non-fermented seaweed and ginger was tested under the same conditions. As a result, In the case of 500 μg / ml of NO, the NO concentration in each comparative group was almost no inhibitory effect of about 60 μM NO concentration compared to 40.6 μM of fermented product. This shows that the present invention can be effectively used for the prevention or treatment of degenerative brain diseases of sewage and ginger fermented products.

Preparation Example 1. Preparation of liquid preparation

Fermented product 10,000 mg

Sugar 200mg

100 mg of isomerized sugar

Lemon incense quantity

Purified water was added to make a total volume of 1000 ml. The above components were mixed according to a conventional method for producing a liquid agent, and then filled in a brown bottle and sterilized to prepare a liquid agent.

Preparation Example 2. Preparation of health drinks

Fermented product 10,000 mg

Citric acid 1000mg

Oligosaccharide 100 g

Plum concentrate 2g

Taurine 1g

Purified water was added to the entire 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the resulting solution was stirred and heated at 85 DEG C for about 1 hour. The resulting solution was filtered and sterilized in a 2 L sterilized container. The resulting solution was refrigerated, ≪ / RTI >

Preparation Example 3. Preparation of health food

5000 mg fermented product

Vitamin mixture quantity

Vitamin A acetate 70 mg

Vitamin E 1.0mg

0.13 mg of vitamin B1

0.15 mg of vitamin B2

Vitamin B6 0.5mg

Vitamin B12 0.2mg

Vitamin C 10mg

Biotin 10mg

Nicotinic acid amide 1.7 mg

Folic acid 50mg

Calcium pantothenate 0.5mg

Vitamin E 1.0mg

Mineral mixture quantity

1.75 mg ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Calcium phosphate diphosphate 55 mg

Potassium citrate 90mg

Calcium carbonate 100 mg

24.8 mg of magnesium chloride

Although the composition ratio of the above-mentioned vitamin and mineral mixture is relatively mixed with a suitable preparation for health food, the compounding ratio may be arbitrarily modified, and the above ingredients may be mixed according to a conventional method of manufacturing health food, The granular formulation of health food was prepared.

While the present invention has been described in connection with what is presently considered to be the preferred embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, .

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. The contents of all publications referred to herein are incorporated herein by reference.

Claims (10)

A pharmaceutical composition for the treatment or prevention of a degenerative brain disease, which comprises a suspension, a ginger, a sugar fermentation product, and a pharmaceutical composition for the treatment or prevention of a degenerative brain disease, wherein the composition comprises 10: 4: 2: .
The pharmaceutical composition for treating or preventing degenerative brain diseases according to claim 1, wherein the fermentation uses Lactobacillus sp. Strain derived from kimchi.
The method according to claim 1,
Wherein said degenerative brain disease is Alzheimer's disease, Parkinson's disease, meningitis, stroke, dementia, Huntington's disease or Creutzfeldt-Jakob disease.
delete Wherein the food composition comprises at least one food ingredient selected from the group consisting of dietary supplements, functional foods, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, Wherein the composition is provided as a food, beverage or food additive.
6. The method of claim 5,
Wherein the composition is fortified with antioxidant components and minerals by fermentation.
A method for producing a sewage and ginger fermentation product,
Mixing ginseng and ginger and sugar in a weight ratio of 10: 4: 2; And
Inoculating the mixture with lactic acid bacterium, and then culturing the mixture at 30 ° C to 40 ° C for 5 to 16 days.
delete 8. The method of claim 7,
Wherein the lactic acid bacterium is a strain of Lactobacillus sp. Derived from kimchi.
8. The method of claim 7,
Wherein the lactic acid bacteria is inoculated at 0.1 CFU / ml to 10.0 CFU / ml, and the culture is performed at 38 ° C for 10 days.

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