KR20090073631A - A composition comprising sulforaphane for preventing and treating cognitive dysfunction - Google Patents

Abstract

A composition for preventing and treating cognitive dysfunction is provided to suppress the nerve cell toxicity which is induced by a beta-amyloid and recover memory damage caused by a scopolamine. A pharmaceutical composition for preventing and treating diseases related to cognitive dysfunction comprises a sulforaphane of the structural formula (I) as an active ingredient. The diseases related to the cognitive dysfunction are alzheimer dementia, intracerbroventricular dementia, Pick disease, Creutzfeldt-jakob disease and Parkinson disease. The compound is contained in the content of 0.1-50 weight%. A health functional food for preventing and treating the diseases related to the cognitive dysfunction comprises the sulforaphane as an active ingredient. The health functional food is used in a form of tablet, capsule, pill and liquid.

Description

A composition comprising sulforaphane for preventing and treating cognitive dysfunction

The present invention relates to a composition for the prevention and treatment of cognitive dysfunction-related diseases containing sulforaphane as an active ingredient.

Selkoe, DJ, Alzheimer's disease: Genes, proteins, and therapy, Physiol Rev , 81 , pp. 741-766, 2001

2, Hardy, JA and Higgins GA, Alzheimer's disease: The amyloid cascade hypothesis, Science , 256 , pp. 184-185, 1992

Markesbery, WR, Oxidative stress hypothesis in Alzheimer's disease, the amyloid cascade hypothesis, Free Radic Biol Med , 23 , pp. 134-147, 1992

[Reference 4] Zhang, Y and Tang, L, Discovery and development of sulforaphane as a cancer chemopreventive phytochemical, Acta Pharmacol Sin , 28 , pp.1343-1354, 2007

[Reference 5] Higdon JV et al . , Cruciferous vegetables and human cnacer risk: epidemiologic evidence and mechanistic basis, Pharmacol Res , 55 , pp.224-236, 2007

Fimognari, C and Hrelia, P, Sulforaphane as a promising molecule for fighting cancer, Mutat Res , 635 , pp. 90-104, 2007

Document 7 Kraft, AD et al . Nuclear factor E2-related factor 2-dependent antioxidant response elemnt activation by tert-butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult, J Neurosci , 24 , pp.1101-1112, 2004

[Reference 8] Zhao, J et al . , Sulforaphane reduces infarct volume following focal cerebral ischemia in rodents, Neurosci Lett , 393 , pp. 108-112, 2006

Document 9 Zhao, J et al ., Sulforaphane enhances aquaporin-4 expression and decreases cerebral edema following traumatic brain injury, J Neurosci Res , 82 , pp. 499-506, 2005

[Document 10] Republic of Korea Patent No. 0497717

[Document 11] Korean Patent Registration No. 0353435

[Document 12] Republic of Korea Patent No. 0737233

[Document 13] Republic of Korea Patent Registration 0773901

Document 14 Jang JH et al . , Ergothioneine rescues PC12 cells from beta-amyloid- induced apoptotic death, Free Radic Biol Med , 36 , pp.288-299, 2004

15. Jang JH and Surh YJ, Protective effect of resveratrol on beta-amyloid-induced oxidative PC12 cell death. Free Radic Biol Med , 34 , pp. 1100-1110, 2003

16. Jang JH and Surh YJ, Possible role of NF-kappa B in Bcl-X _L protection against hydrogen peroxide-induced PC12 cell death. Redox Rep , 9 , pp.343-348, 2004

Document 17 Kazue T et al . , Effect of selegiline alone or with donepizil on memory impairmaent in rats, Eur Pharmacol , 518 , pp. 140-144, 2005

Dementia, a representative chronic progressive degenerative brain disease, affects both mental cognition and social behavior, resulting in normal daily life disorders. It is a symptomatic complex that refers to the case where these criteria are met. There are three major types of dementia, 1) Alzheimer's disease (AD), in which cognitive deterioration progresses without any obvious cause, and 2) vascular dementia caused by cerebrovascular diseases such as repeated cerebral infarction, Third, it is classified as dementia caused by other diseases. Cerebrovascular dementia is mainly caused by atherosclerosis, and the symptoms can be improved by treatment with thrombolytics, but Alzheimer's disease has not been fully identified.

The main parts of Alzheimer's disease are the hippocampus, forebrain, and temporal lobe. As the disease progresses, about 30-40% of the nerves are lost. To date, Alzheimer's disease has been pathologically characterized by cerebral atrophy in the cerebral cortex or hippocampus and the deposition of senile plaques or neurofibrillary tangles in the brain (Selkoe DJ, Alzheimer's). disease: Genes, proteins, and therapy, Physiol Rev , 81 , pp. 741-766, 2001). Observation of the development of Alzheimer's disease revealed the accumulation of a protein called beta amyloid (β), followed by dementia and neurofibrillary tangles. The senile plaques are structures of neutrophil spots seen in the cerebral cortex and hippocampus of the brain, with amyloid fibers in the center and surrounding degenerated neuronal processes. In other words, beta amyloid is a major component of the senile plaque accumulated in the brain of Alzheimer's disease patients, and it is thought that the toxicity of this substance to neurons is a major cause of Alzheimer's disease (Hardy, JA and Higgins GA, Alzheimer's). disease: The amyloid cascade hypothesis, Science , 256 , pp. 184-185, 1992).

The mechanisms by which over-produced beta amyloid causes toxicities vary, but the following two theories can explain disease progression. The first theory is that amyloid plaques induce microglia, which in turn trigger a series of inflammatory reactions, including damaging nerves and stimulating the production of cytokines that attack. The second theory is based on the action of reactive oxygen species. More than 95% of the brain is made up of fat and is oxidized under the attack of free radicals. Beta amyloid forms reactive oxygen species (ROS) such as hydrogen peroxide, superoxide anion, and hydroxylradical through aggregation regulation, which affects macromolecules in vivo due to its high reactivity and ultimately active cells in neurons Causes death (Markesbery, WR, Oxidative stress hypothesis in Alzheimer's disease, The amyloid cascade hypothesis, Free Radic Biol Med , 23 , pp. 134-147, 1992).

On the other hand, in general, broccoli belongs to sipjagwa plant (cruciferous vegetable), cabbage, cauliflower, etc. a number of in to have, and it contains highly various physiologically active substances, including flavonoids, of broccoli has excellent efficiency in cancer prevention and treatment in vitro and in Reported in vivo studies (Zhang, Y and Tang, L, Discovery and development of sulforaphane as a cancer chemopreventive phytochemical, Acta Pharmacol Sin , 28 , pp. 1343-1354, 2007). Epidemiologic studies have shown broccoli to reduce the risk of breast, lung, colorectal and prostate cancers (Higdon JV et al . , Cruciferous vegetables and human cnacer risk: epidemiologic evidence and mechanistic basis, Pharmacol Res , 55 , pp. 224-236, 2007). In particular, cruciferous plants contain large amounts of glucosinolates and their hydrolysates, indole and isothiocyanate, and the representative ingredient is sulforaphane. Sulforapane has antioxidant and anti-inflammatory effects, and acts on a multi-stage process of cancer initiation, promotion, and progression, showing anticancer efficacy (Fimognari, C and Hrelia, P, Sulforaphane as a promising molecule for fighting cancer, Mutat Res , 635 , pp. 90-104, 2007). At the onset of cancer, the procarcinogen is metabolized to inhibit the conversion of the final carcinogen to the ultimate carcinogen, and the in vivo phase I and phase II enzymes are activated to detoxify the xenobiotic. Ultimately, it inhibits gene damage caused by carcinogens. It is known that the promotion phase induces apoptosis of cancer cells and inhibits the cell cycle. In its progression, it has been reported to have protective effects against angiogenesis, malignant transformation, invasion and metastasis.

Compared to the anti-cancer effects of sulforaphane, the study of sulforaphane's action in the brain is still in its infancy. Some protective actions reported in the brain include: in In vitro neuronal cell culture model inhibited the cytotoxicity caused by hydrogen peroxide, a typical reactive oxygen species, and glutamic acid, a neurotoxic substance (Kraft, AD et. al . Nuclear factor E2-related factor 2-dependent antioxidant response elemnt activation by tert-butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult, J Neurosci , 24 , pp.1101-1112, 2004), in vivo It has been reported to have protective effects against brain injury due to ischemia and trauma in experimental animal models (Zhao, J et. al . , Sulforaphane reduces infarct volume following focal cerebral ischemia in rodents, Neurosci Lett , 393 , pp. 108-112, 2006; Zhao, J. et al ., Sulforaphane enhances aquaporin-4 expression and decreases cerebral edema following traumatic brain injury, J Neurosci Res , 82 , pp. 499-506, 2005).

      On the other hand, all of the prior patents related to broccoli and sulforaphane are not related to dementia or memory enhancement at all, and thus broccoli somatic embryo culture method and mass production method of broccoli plant sprouts using the same (Korea Patent No. 0497717), containing broccoli vegetable Functional beverage and its manufacturing method (Korean Patent No. 0353435), a method of preparing kimchi based on broccoli (Korean Patent No. 0737233), and a method of producing lactic acid bacterium fermented milk having inhibitory effect of Helicobacter pylori using broccoli sprout extract Most of these are simple patents, such as (Korean Patent No. 0773901). However, the patents are not related to the preventive treatment and memory enhancement effect of dementia, and there was no technology for making broccoli and sulforaphane for dementia prevention and memory enhancement, or a pharmaceutical composition. .

     Accordingly, the present inventors have completed the present invention by confirming that the composition containing sulforaphane as an active ingredient has an excellent protection against beta amyloid-induced cytotoxicity and scopolamine-induced memory damage.

In order to achieve the above object, it provides a pharmaceutical composition for the prevention and treatment of cognitive dysfunction-related diseases containing a sulforaphane represented by the following structural formula (I) as an active ingredient.

In another aspect, the present invention provides a health functional food for the prevention and improvement of cognitive impairment-related diseases containing a sulforaphane represented by the following structural formula (I) as an active ingredient.

(Ⅰ)

(Ⅰ)

The compound is separable from cruciferous plant extracts or commercially available.

The cognitive dysfunction related diseases include Alzheimer's dementia, cerebrovascular dementia, pick disease, Creutzfeldt-jakob disease, dementia due to head injury or Parkinson's disease, preferably Includes Alzheimer's disease.

The composition of the present invention comprises 0.1 to 50% by weight of the compound relative to the total weight of the composition.

The present invention provides a pharmaceutical composition containing the compound as an active ingredient.

Compositions comprising a compound of the present invention may further comprise a suitable carrier, excipient or diluent according to conventional methods.

Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The compositions comprising the compounds of the present invention are each formulated in the form of oral dosage forms, external preparations, suppositories, or sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., in accordance with conventional methods. Can be used.

Specifically, when formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose, etc. ), Lactose, gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc can also be used. Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The compound according to the present invention may vary depending on the age, sex and weight of the patient, but is generally administered in an amount of 0.01 to 500 mg / kg, preferably 0.1 to 100 mg / kg, divided once or several times daily. can do. In addition, the dosage of the compound may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Since the compound of the present invention has little toxicity and side effects, it can be used with confidence even for long-term administration for the purpose of prevention.

The present invention provides a health functional food for the prevention and improvement of cognitive dysfunction-related diseases containing the compound as an active ingredient.

The composition comprising the compound of the present invention may be used in various ways, such as drugs, foods and drinks for the prevention of cognitive dysfunction-related diseases.

Foods to which the compound of the present invention may be added include various foods, for example, beverages, gums, teas, vitamin complexes, dietary supplements, and the like, which are pills, powders, granules, tablets, tablets, capsules or beverages. Available in form.

At this time, the amount of the compound in the food or beverage, in the case of the health food composition of the present invention can generally be added to 0.01 to 15% by weight of the total food weight, 0.02 to 10 g, based on 100 ml for the health beverage composition, Preferably it can be added in the ratio of 0.3-1 g.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the compound as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Polysaccharides such as dextrin, cyclodextrin; Conventional sugars such as and the like and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 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 includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention is useful as a pharmaceutical composition and health functional food for the prevention and treatment of cognitive dysfunction-related diseases by confirming the effect of sulforaphane on beta amyloid-induced neuronal toxicity and scopolamine-induced memory damage. It is available.

Hereinafter, the present invention will be described in detail by the following reference examples, examples and experimental examples.

However, the following Reference Examples, Examples, and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited thereto.

Reference Example  1. Experiment Preparation and Reagents

1-1. Cell culture

Neuronal SH-SY5Y neuroblastoma cell line was purchased from ATCC (RL-2266) and mixed with DMEM culture with 10% FBS (14-501FM, Cambrex, USA), penicillin and streptomycin (15140-122, Gibco BRL, USA). (12800-17, Gibco BRL, USA) was incubated under 37 ℃, 5% CO ₂ conditions, the medium was changed once every two days.

1-2. Reagent Preparation

Beta amyloid (Aβ _25-35 : Bachem, USA) was dissolved in distilled water to make 1 mM and stored at -20 ℃, each time diluted in DMEM medium to a final concentration of 25 μM each time. Sulforapane is manufactured by LKT Laboratories, Inc. It was purchased from (USA) and dissolved in DMSO (dimetylsulfoxide) to make a stock (stock) solution of the concentration of 1, 2, 5 mM and diluted with DMEM to the required concentration. Cells were incubated for 30 minutes with various concentrations of sulforaphane, followed by further beta amyloid, followed by further incubation at 37 ° C for 24 hours.

1-3. Experimental animals

As experimental animals, Spraque-Dawley male rats (Hyochang Science, Daejeon) were weighed about 250 g and used for one week. The test and control groups were divided into three groups of five animals each. During the adaptation period and the experiment period, water and feed were freely eaten. The temperature in the experimental animal breeding room was maintained at 21-26 ℃ and humidity 40-60%, and 12 hours and 12 hours were performed during the day.

Experimental Example 1. Effect of sulforaphane cell viability

In order to confirm the cell viability effect of the sulforaphane of Reference Example 1-2, the following experiment was applied by applying the method described in the existing literature (Jang JH et. al . , Ergothioneine rescues PC12 cells from beta-amyloid-induced apoptotic death, Free Radic Biol Med, 36 , pp.288-299, 2004).

For cytotoxicity analysis, 48-well microtiter plates were dispensed at a cell concentration of 6 × 10 ⁴ cells / 300μl and incubated for 24 hours.The sulforaphane was diluted to 1, 2 or 5μM. 30 minutes later, beta amyloid was treated. After incubation for 24 hours, 10 μl of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) was added and reacted for 2 hours to form formazan. The supernatant was removed and dissolved in formazan by adding 200 μl / well of DMSO, and then measured at a wavelength of 570 nm using an ELISA reader (Emax, Molecular device, USA). It was confirmed that the cytotoxicity induced by beta amyloid is reduced (see FIG. 1).

Experimental Example 2. Reducing the free radical species (ROS) of sulforaphane

In order to confirm the active oxygen species reduction effect of the sulforaphane of Reference Example 1-2, the following experiment was applied by applying the method described in the existing literature (Jang JH and Surh YJ, Protective effect of resveratrol on beta-amyloid-induced oxidative PC12 cell death. Free Radic Biol Med , 34 , pp. 1100-1110, 2003).

The sulforaphane was pretreated for 30 minutes by concentration, incubated for 6 hours with the addition of 25 μM of beta amyloid, the cells were washed twice with Krebs-Ringer solution, and then reacted for 15 minutes at 37 ° C. with 10 μM DCF-DA. Fluorescence changes were measured with a fluorometer (GeminiXS, Molecular Device, USA) at excitation wavelength 485 nm and emission wavelength 535 nm. Lipid-soluble DCF-DA enters the cell and undergoes esterase or oxidative hydrolysis to deacetylate non-fluorescent DCFH, which is oxidized by active oxygen to become a strong fluorescence DCF. As a result, as shown in Figure 2, it was confirmed that the active oxygen species induced by beta amyloid at 5 μM sulforaphane markedly reduced (see Fig. 2).

Experimental Example  3. Sulforaphane  Antioxidant enzymes and neurons Protector  Increased effect of expression

In order to confirm the effect of increasing the expression of antioxidant enzymes and neuroprotective factors of the sulforaphane of Reference Example 1-2, the following experiment was applied by applying the method described in the existing literature (Jang JH and Surh YJ, Possible role of NF _{-kappaB in Bcl-X L protection against} hydrogen peroxide-induced PC12 cell death. Redox Rep, 9, pp.343-348, 2004).

Total RNA was extracted and separated by TRI reagent (Molecular Research, Inc., USA) on the cultured cells of Reference Example 1-1, chloroform was added and mixed, followed by centrifugation at 12,000 rpm for 15 minutes. The upper layer containing RNA was separated and the RNA was precipitated with isopropyl alcohol. The RNA pellet was washed with 75% ethanol, dried, dissolved in RNase free water, converted to cDNA using M-MLV reverse transcriptase (Promega, USA), and then used for each primer corresponding to a specific DNA sequence. Amplified by PCR reaction solution. The product was electrophoresed on 1.5% agarose gel and stained with EtBr (ethidium bromide) and photographed by UV microscope (GelDoc, TCP-20.M, Vilber Lourmat, France).

Experimental results, as shown in Figure 3A treated with sulforaphane 5μM alone and confirmed after 0, 1, 3, 6, 12, 24 hours, it was confirmed that the mRNA expression of the representative antioxidant enzyme, catalase increased, and in Figure 3B As shown, when sulforaphane 5μM pretreatment for 30 minutes and beta amyloid 25μM for 24 hours, it was confirmed that the expression of beta amyloid reduced antioxidant enzymes increased sulforaphane concentration dependent (see Figure 3).

In addition, mNRA expression of neuronal cell protection and growth factor brain-derived neurotrophic factor (BDNF) as shown in Figure 4 also confirmed that the time-dependent increase by the treatment of sulforaphane (see Figure 4).

Experimental Example  4. With scopolamine  Inhibition of induced memory impairment

In order to confirm the scopolamine-induced memory damage inhibition effect of the sulforaphane of Reference Example 1-2, the following experiment was applied by applying the method described in the existing literature (Kazue T et. al . , Effect of selegiline alone or with donepizil on memory impairmaent in rats, Eur Pharmacol , 518 , pp. 140-144, 2005).

Scopolamine is known to act as an acetylcholine receptor antagonist to inhibit memory. Three days prior to scopolamine administration, sulforaphane was intraperitoneally administered at a dose of 5 mg / kg and physiological saline was administered to the control group. Thereafter, 0.5 mg / kg of scopolamine was intraperitoneally administered for 5 days to cause memory impairment, and a water-maze test was performed three times daily for 5 days. Sumi-ro inspection includes a cylindrical water tank (180 cm in diameter and 60 cm in height) with a platform (platform: 12 cm in diameter and 35 cm in height) and a video-tracking system (Ethovision, Noldus) for recording the rat's movements. , The Netherlands). At this time, the water temperature was 21 ± 2 ℃, the water was up to 2 cm above the platform. When the rats visited the platform and stayed at the platform for more than 30 seconds, the time taken until the visit was set as escape latency, and the average value of the three times a day was used as mean escape latency. At this time, if the average escape latency exceeds 90 seconds, the same value was taken as 90 seconds. Therefore, a total of 15 experiments were conducted per rat during the 5-day irradiation period.

As a result, the control group not administered scopolamine significantly decreased the time to visit the platform as the training period was longer, and the rats who received scopolamine only intraperitoneally could hardly find the platform regardless of the training period. It was confirmed that the rats intraperitoneally administered at 5 mg / kg / day sulforaphane reduced the time to visit the platform on the 4th and 5th day of the experimental period (see FIGS. 5A and 5B).

Experimental Example  5. Cytotoxicity Experiment

In order to confirm the cytotoxicity test of sulforaphane of Reference Example 1-2, the following experiment was applied by applying the method described in the existing literature (Jang JH et. al . , Ergothioneine rescues PC12 cells from beta-amyloid-induced apoptotic death, Free Radic Biol Med , 36 , pp. 288-299, 2004).

For cytotoxicity analysis, the cells were dispensed at a cell concentration of 6 x 10 ⁴ cells / 300μl in a 48-well microtiter plate and incubated for 24 hours.The concentration of sulforaphane was 1, 2 and 5 μM. After diluting with, incubated for 24 hours, 10 μl of MTT solution was added and reacted for 2 hours to form formazan. The supernatant was removed, DMSO 200 μl / well was added to dissolve formazan, and then ELISA. As a result of measurement at a wavelength of 570 nm using a reader (Emax, Molecular device, USA), as shown in Figure 6 it was confirmed that there is little toxicity of the cells (see Figure 6).

Thus, the preparation examples using the compounds below, but this is intended to explain in detail only, not intended to limit the present invention.

Formulation example  One. Powder  Produce

Sulforaphane 300 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

Sulforaphane 300 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation example  3. Manufacture of capsule

Sulforaphane 300 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation example  4. Preparation of Injectables

Sulforaphane 300 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation example  5. Liquid  Produce

Sulforaphane 300 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled into a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of healthy food

Sulforaphane 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

       Sulforaphane 300 mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantification

After mixing the above components in accordance with a conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

1 is a diagram showing the effect of reducing sulforaphane beta amyloid-induced toxicity in SH-SY5Y neuroblastoma cells,

2 is a diagram showing the effect of reducing the active oxygen species caused by beta amyloid sulforaphane,

Figure 3 is a diagram showing the effect of increasing the expression of catalase is a representative antioxidant enzyme in the cell sulforaphane,

4 is a diagram showing the effect of sulforaphane increases the expression of neuronal growth factor,

5 is a diagram showing a recovery effect of memory impairment when sulforaphane is administered to rats whose memory is impaired with scopolamine.

Figure 6 shows the toxicity of sulforaphane to SH-SY5Y neuroblastoma cells, sulforaphane is a diagram showing that almost no cytotoxicity in the concentration range of 5 μM or less for neurons.

Claims (5)

A pharmaceutical composition for the prevention and treatment of cognitive dysfunction-related diseases containing sulforaphane represented by the following structural formula (I) as an active ingredient:

(Ⅰ) The method of claim 1, wherein the cognitive impairment-related disease is Alzheimer's dementia, cerebrovascular dementia, pick disease, Creutzfeldt-jakob disease, dementia due to head injury, or Parkinson's disease. Pharmaceutic composition. The pharmaceutical composition according to claim 1, comprising 0.1 to 50% by weight of the compound relative to the total weight of the composition. A health functional food for the prevention and improvement of cognitive impairment-related diseases containing the sulforaphane represented by the structural formula (I) of claim 1 as an active ingredient. The dietary supplement of claim 4 which is a tablet, capsule, pill or liquid.

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