WO2025170200A1 - Novel composition comprising wheat germ extract - Google Patents

Abstract

The present invention relates to a composition comprising a wheat germ extract and, more specifically, to a composition comprising a wheat germ extract as an active ingredient for the prevention, alleviation, or treatment of neurodegenerative diseases, aging-related diseases, or hangovers. In addition, the composition of the present invention may further comprise at least one selected from the group consisting of a chamomile extract and a Sophora japonica extract.

Description

Novel composition comprising wheat germ extract

The present invention relates to a composition comprising a wheat germ extract, and more particularly, to a composition comprising a wheat germ extract as an active ingredient for preventing, improving, or treating neurodegenerative diseases, age-related diseases, or hangovers. Furthermore, the composition of the present invention may further comprise at least one selected from the group consisting of a chamomile extract and a pine extract.

With the recent rapid increase in the elderly population and the increase in patients with various age-related diseases and neurodegenerative diseases, interest in their treatment and prevention is growing.

Aging is the progressive loss and deterioration of function at the cellular, tissue, and organ levels, leading to a progressive loss of physiological integrity, increased susceptibility to disease and external stressors, and ultimately death. As the global population ages, the incidence of age-related diseases is increasing annually. Consequently, numerous attempts have been made to treat age-related diseases and delay the onset of the complex aging process. Consequently, numerous aging-related pathways have been identified that can be targeted to extend lifespan and healthspan. This raises hopes for novel interventions, including drugs that slow the aging process and modulate conserved aging pathways to delay the onset of age-related diseases. However, to date, aside from some symptomatic treatments, no known interventions have been shown to effectively slow the human aging process. Consequently, beyond pharmacological treatments for existing diseases and disorders, there is a growing need and demand for strategies to maintain health and delay aging.

Neurodegenerative diseases are conditions that cause various pathologies, such as movement disorders, memory impairment, and cognitive impairment, due to the decline or loss of nerve cell function. Nerve cells die in large numbers every day not only in neurological diseases but also in the brains of normal adults, and the number of nerve cells that die increases exponentially with aging. Major neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, and Huntington's disease, and the pathogenesis of these diseases is still not fully understood. Acetylcholinesterase inhibitors or NMDA (N-methyl-D-aspartate) receptor antagonists are used to treat Alzheimer's disease, while L-dopa, dopamine agonists, MAO-B inhibitors, or COMT inhibitors are used to treat Parkinson's disease. Dopamine D2 receptor antagonists are used to treat Huntington's disease. However, since these treatments all target neurotransmission processes, their use only alleviates symptoms, not fundamental cures. Therefore, there is a continuous need for new drugs that can provide fundamental treatment.

Meanwhile, modern people are trying to relieve stress in various ways. One such method is drinking. In South Korea, excessive drinking culture is a problem compared to other countries, and hangover cures are often taken after drinking to alleviate hangover symptoms.

SIRT1 (silent mating type information regulation 2 homolog; sirtuin 1) is an NAD+-dependent deacetylase known to regulate protein function by deacetylating lysine residues in various proteins. It is most similar to yeast Sir2, which has NAD+-dependent class ² histone deacetyl activity. SIRT1 is known to be a regulator involved in various physiological activities. For example, it is involved in activities such as genome stabilization, cancer, stress response, apoptosis, metabolism, obesity, diabetes, and aging, and it has also been confirmed to be involved in neuroinflammation and neurodegeneration in the nervous system. Specifically, SIRT1 expression is known to affect the activity of Alzheimer's disease and Huntington's disease (see Non-Patent Documents 1 and 2).

Furthermore, the SIRT1 protein plays a crucial role in protecting the liver from alcohol-induced oxidative stress. Ethanol consumption inhibits SIRT1 activity, leading to fat accumulation and inflammation. SIRT1 regulates lipids and fatty acids through the AMPK, PPAR-α, and PGC-1α pathways. Alcohol disrupts these pathways, increasing fat accumulation and reducing fatty acid oxidation. SIRT1 activation can reverse alcohol-induced liver damage.

Additionally, SIRT1 plays an important role in alleviating neuroinflammation, and may reduce cell damage in brain injury or neurodegenerative diseases by suppressing the activation of microglia (the main immune cells in the brain) and reducing neuroinflammatory responses through a pathway that inhibits inflammatory proteins and inflammasome activation.

Wheat ( Triticum vulgare ) is an important food crop, one of the world's three major crops, along with rice and barley. The wheat germ, which accounts for 2-3% of the grain, contains 23 highly concentrated nutrients, providing essential nutrients for wheat growth. However, wheat germ is often discarded industrially during the milling process to produce flour because it contributes to rancidity and a bitter taste.

The inventors of the present invention have completed the present invention by confirming that a composition comprising wheat germ extract and chamomile extract has excellent effects on the human body, specifically, excellent effects in improving, treating or preventing neurological and aging-related diseases, and excellent effects in relieving and preventing hangovers.

(Prior art literature)

(Patent Document)

(Patent Document 0001) Registered Patent Publication No. 10-2217039

(Non-patent literature)

(Non-patent document 0001) Jessica A. Hall et al., The sirtuin family's role in aging and age-related pathologies, J Clin Invest. 2013;123(3):973-979.

(Non-Patent Document 0002) Fangzhou Jiao and Zuojiong Gong, The Beneficial Roles of SIRT1 in Neuroinflammation-Related Diseases, Oxidative Medicine and Cellular Longevity Volume 2020, Article ID 6782872.

The present invention provides a composition comprising a substance that increases the activity of the SIRT1 gene or the expression of the SIRT1 gene.

In addition, the present invention provides a composition used for preventing or improving neurodegenerative diseases, aging-related diseases, or hangovers by increasing the activity of the SIRT1 gene or the expression of the SIRT1 gene.

The present invention comprises (1) quinone or wheat germ extract; and

(2) a composition comprising at least one selected from the group consisting of apigenin or chamomile extract, and (3) quercetin or chamomile extract.

In one embodiment, the present invention relates to a composition comprising (1) quinone or wheat germ extract and (2) apigenin or chamomile extract.

In one embodiment, the present invention relates to a composition comprising (1) quinone or wheat germ extract and (3) quercetin or syringa extract.

In one embodiment, the present invention relates to a composition comprising (1) quinone or wheat germ extract; (2) apigenin or chamomile extract; and (3) quercetin or chamomile extract.

In one embodiment, the present invention relates to a composition comprising wheat germ extract, chamomile extract, and pine tree extract.

In one embodiment, the present invention relates to a composition comprising quinone, apigenin and quercetin.

The above quinone may be at least one selected from the group consisting of 2,6-DMBQ (2,6-dimethoxy-1,4-benzoquinone), 2-MBQ (2-methoxy-1,4-benzoquinone), and rapachone, but is not limited thereto.

In addition, the wheat germ extract, chamomile extract, and pine tree extract of the present invention can be extracted with a solvent selected from the group consisting of water, C1 to C6 alcohols, and mixed solvents thereof. Preferably, the wheat germ extract, chamomile extract, and pine tree extract may be 0.01% to 99% ethanol extracts. More preferably, the wheat germ extract, chamomile extract, and pine tree extract may be 50% to 70% ethanol extracts. Most preferably, the wheat germ extract, chamomile extract, and pine tree extract may be 60% ethanol extracts.

The composition of the present invention may be a mixture of two or more extracts selected from the group consisting of wheat germ, chamomile, and cinnamon tree, or a mixture of extracts of each of these.

The composition of the present invention can increase SIRT1 gene activity or SIRT1 gene expression.

The composition of the present invention can prevent or treat age-related diseases. The age-related diseases may include geriatric cardiovascular diseases and disorders, geriatric lung diseases, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoarthritis, geriatric ocular diseases, or skin aging.

The composition of the present invention can prevent or treat neurodegenerative diseases. The neurodegenerative diseases may be stroke, cerebral infarction, cerebral embolism, ischemic neurological disease, degenerative neurological disease, apoplexy, Alzheimer's disease, Parkinson's disease, dementia, Huntington's disease, Pick's disease, Creutzfeldt-Jakob disease, multiple sclerosis, ischemic encephalopathy, epilepsy, Lou Gehrig's disease, memory loss, nerve cell damage due to trauma or disease, concussion, progressive supranuclear palsy, multiple system atrophy, olivary-pontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, amyotrophic lateral sclerosis (ALS), essential tremor, corticobasal degeneration, diffuse Lewy body disease, Parkinson's-ALS-dementia complex, or Friedreich's ataxia.

The composition of the present invention can prevent or treat hangovers. The composition of the present invention can prevent, improve, or treat hangovers by decomposing blood alcohol or acetaldehyde.

In one embodiment, the composition of the present invention relates to a food composition that prevents or improves an aging-related disease, a neurodegenerative disease, or a hangover.

In one embodiment, the composition of the present invention relates to an animal feed composition that prevents or improves aging-related diseases, neurodegenerative diseases, or hangovers.

The present invention relates to a composition comprising wheat germ, chamomile, and sycamore extracts as active ingredients, wherein the composition can increase the activity or expression of the SIRT1 gene. Accordingly, the composition of the present invention can be provided as a pharmaceutical composition, a food composition, or an animal feed composition.

Therefore, the composition of the present invention can be usefully used for the prevention, improvement, or treatment of neurodegenerative diseases, aging-related diseases, or hangovers by increasing the activity or expression of the SIRT1 gene.

Figure 1 shows the concentration of blood alcohol when wheat germ extract, chamomile extract, and a mixture thereof were administered to rats administered ethanol.

Figure 2 shows the concentration of acetaldehyde in blood when wheat germ extract, chamomile extract, and a mixture thereof were administered to rats administered ethanol.

Figure 3 shows the NAD+/NADH ratio in hepatocytes when treated with wheat germ extract, chamomile extract, ash tree extract, and a mixture thereof.

Figure 4 shows the sirt1 expression ratio in hepatocytes when treated with wheat germ extract, chamomile extract, pine tree extract, and a mixture thereof.

Figure 5 shows the degree of inflammation inhibition when wheat germ extract, chamomile extract, pine tree extract, and a mixture thereof are treated in nerve cells.

Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the attached drawings so that those skilled in the art can easily implement the present invention. However, this specification may be implemented in various forms and is not limited to the embodiments and examples described herein.

Throughout this specification, whenever a part is said to "include" a component, this means that it may include other components, but not to the exclusion of other components, unless otherwise stated.

The present invention comprises (1) quinone or wheat germ extract; and

(2) a composition comprising at least one selected from the group consisting of apigenin or chamomile extract, and (3) quercetin or chamomile extract.

The present invention also relates to a composition comprising (1) quinone or wheat germ extract and (2) apigenin or chamomile extract.

In addition, the present invention relates to a composition comprising (1) quinone or wheat germ extract and (3) quercetin or syringa extract.

The present invention relates to a composition comprising (1) quinone or wheat germ extract; (2) apigenin or chamomile extract; and (3) quercetin or chamomile extract.

The present invention relates to a composition comprising wheat germ extract, chamomile extract and pine tree extract.

The present invention relates to a composition comprising quinone, apigenin and quercetin.

The component extracted from wheat germ may be one or more quinones selected from the group consisting of, but not limited to, 2,6-DMBQ (2,6-dimethoxy-1,4-benzoquinone), 2-MBQ (2-methoxy-1,4-benzoquinone) and lapachone.

The above quinones each have the following chemical structure.

[2,6-DMBQ (2,6-dimethoxy-1,4-benzoquinone)]

[2-MBQ (2-methoxy-1,4-benzoquinone)]

[Lapachone]

The quinone of the present invention can be extracted from wheat germ. In the present invention, the component extracted from wheat germ may be provided in the form of a purified compound or in the form of a wheat germ extract.

Apigenin can be extracted from chamomile. Apigenin has the following chemical structure.

[Apigenin (4', 5, 7-trihydroxyflavone)]

Quercetin can be extracted from the quercetin tree. The quercetin has the following chemical structure.

[Quercetin]

The composition according to the present invention may be a pharmaceutical composition, a food composition or an animal feed composition.

The extract used in the present invention can be obtained using a conventional extraction solvent known in the art. The extraction solvent may be a polar solvent or a non-polar solvent. Polar solvents include, but are not limited to, water, C1 to C6 alcohols (e.g., methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, and n-butanol), acetic acid, or a mixture of the above polar solvents. Non-polar solvents include, but are not limited to, acetone, acetonitrile, ethyl acetate, methyl acetate, butyl acetate, fluoroalkane, hexane, ether, chloroform, dichloromethane, or a mixture of the above non-polar solvents.

The extract used in the present invention may be extracted through hot water extraction, cold extraction, reflux cooling extraction, ultrasonic extraction, or a conventional extraction method known in the art.

The term "extract" used in the present invention refers to what is commonly referred to as a crude extract in the art, but broadly includes fractions obtained by further fractionating the extract. That is, the extract includes not only those obtained using the above-mentioned solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity), and other additionally performed fractions obtained through various purification methods are also included in the extract of the present invention.

SIRT1 (silent mating type information regulation 2 homolog; sirtuin 1) is known as NAD+-dependent deacetylase Sirtuin-1, and is also referred to as SIR2L1, SIR2, hSIR2, SIR2alpha, Sirtuin 1, etc.

The food composition of the present invention may be a health functional food, dairy product, fermented product, or food additive. The health functional food refers to a food manufactured and processed using raw materials or ingredients with beneficial functions for the human body. "Functionality" refers to ingestion for the purpose of obtaining beneficial effects for health purposes, such as regulating nutrients for the structure and functions of the human body or physiological functions.

In the present invention, “treatment” means partially or completely alleviating, improving, alleviating, inhibiting or delaying a symptom of a specific disease, disorder and/or condition or condition, reducing the severity or reducing the occurrence of one or more symptoms or characteristics.

In the present invention, “prevention” means any action that suppresses or delays the onset of a specific disease, and “improvement” means any action that at least reduces a parameter related to the condition being treated, for example, the degree of symptoms.

The present invention will be described in more detail through the following examples; however, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[Manufacturing Example 1]

Preparation of wheat germ extract

Wheat germ extract was prepared by extracting wheat germ with 60% ethanol at room temperature for 18 hours, filtering and concentrating the extract, and then spray drying it.

The "wheat germ" of the present invention is the nutrient-rich germ of the wheat kernel, removed during the process of processing whole wheat into flour, and accounts for approximately 2-3% of the total wheat kernel. Wheat germ, the germ of wheat, is known to be rich in highly concentrated nutrients and vitamin E (tocopherol), an antioxidant vitamin.

[Manufacturing Example 2]

Preparation of chamomile extract

Chamomile extract was prepared by extracting chamomile with 60% ethanol at room temperature for 18 hours, filtering and concentrating it, and then spray drying it.

The "chamomile ( Chamomilla recutita )" of the present invention is a herb belonging to the dicotyledonous plant family Asteraceae, and is also called chamomile, camomile, or camomile. Chamomile is known to have an excellent calming effect, helping to relieve various pains, and to have the effect of soothing skin troubles.

[Manufacturing Example 3]

Preparation of Conversation Tree Extract

The fruit of the pine tree was extracted with 60% ethanol at room temperature for 2 hours, filtered and concentrated, and then spray-dried to produce a pine tree extract.

The " Sophora japonica L " of the present invention is a deciduous broad-leaved tree belonging to the legume family, utilized for various purposes such as medicinal, ornamental, and timber purposes, and is known to be particularly effective in alleviating menopausal symptoms. The Sophora japonica L fruit extract is a menopausal functional raw material approved by the Ministry of Food and Drug Safety, and is rich in sophoricoside, a type of isoflavone.

[Manufacturing Example 4]

Preparation of a mixture of wheat germ extract and chamomile extract

A mixture was prepared by mixing the wheat germ extract of Manufacturing Example 1 and the chamomile extract of Manufacturing Example 2 in a ratio of 4:1 (w/w) or 1:2 (w/w).

[Manufacturing Example 5]

Preparation of a mixture of wheat germ extract, chamomile extract, and pine tree extract

A mixture was prepared by mixing the wheat germ extract of Manufacturing Example 1, the chamomile extract of Manufacturing Example 2, and the pine tree extract of Manufacturing Example 3 in a ratio of 4:1:1 (w/w) or 1:2:2 (w/w).

[Manufacturing Example 6]

Preparation of a mixture of wheat germ extract, apigenin, and quercetin

A composition was prepared by mixing the wheat germ extract, apigenin, and quercetin of Manufacturing Example 1 in the weight ratio shown in Table 1 below.

[Manufacturing Example 7]

Preparation of a mixture of quinone, apigenin, and quercetin

A composition was prepared by mixing quinone, apigenin, and quercetin in the weight ratios shown in Table 2 below. DMBQ (2,6-dimethoxy-1,4-benzoquinone), 2-MBQ (2-methoxy-1,4-benzoquinone), or Lapachone was used as the quinone.

[Experimental Example 1]

Evaluation of SIRT1 expression in compositions containing wheat germ extract

An experiment was conducted to evaluate the SIRT1 expression level for Examples 1 to 3 and Comparative Examples 4 to 6 manufactured according to Manufacturing Example 6. Comparative Examples 4 to 6 are each a single composition of wheat germ extract, quercetin, and apigenin, as shown in Table 3 below.

Specifically, C2C12 cells were seeded on a 60 mm plate in DMEM medium, and when the cells grew to about 80%, a control group in which the cell line was not treated with a test substance and a group in which the same cell line was treated with Examples 1 to 3 and Comparative Examples 4 to 6, respectively, were cultured for 24 hours.

Afterwards, cells were collected and SIRT1 protein was quantified through Western blotting. The relative expression level of Sirt1 protein was calculated, and the results are shown in Table 3 below.

As shown in Table 3 above, among the comparative examples in which wheat germ extract, apigenin, and quercetin were each treated, the SIRT1 expression level of Comparative Example 4, which was a single treatment group of wheat germ extract, was the highest.

However, Examples 1 to 3, which are mixed compositions, showed higher SIRT1 expression levels compared to Comparative Examples 4 to 6, which are single compositions. In particular, Example 1, which included all of wheat germ extract, apigenin, and quercetin, was confirmed to have a SIRT1 expression level that was 30% higher than that of Comparative Example 4, which was a single wheat germ extract treatment group.

Accordingly, it was confirmed that the composition containing the wheat germ extract of the present invention has a high level of SIRT1 expression, and furthermore, when apigenin and quercetin are individually or mixed and treated with the wheat germ extract, the level of SIRT1 expression increases, and it can be seen that it can exhibit excellent effects on neurodegenerative diseases, aging-related diseases, and hangovers.

[Experimental Example 2]

Evaluation of SIRT1 expression in compositions containing quinone

An experiment was conducted to evaluate the SIRT1 expression level for Examples 4 to 12 and Comparative Examples 1 to 3 manufactured according to Manufacturing Example 7 above. Comparative Examples 1 to 3 are each a single composition of DMBQ (2,6-dimethoxy-1,4-benzoquinone), 2-MBQ (2-methoxy-1,4-benzoquinone), or Lapachone as a quinone, as shown in Table 4 below.

Specifically, C2C12 cells were seeded on a 60 mm plate in DMEM medium, and when the cells grew to about 80%, a control group in which the cell line was not treated with a test substance and a group in which the same cell line was treated with Examples 4 to 12 and Comparative Examples 1 to 3 were cultured for 24 hours.

Afterwards, cells were collected and SIRT1 protein was quantified via Western blotting. The relative expression level of Sirt1 protein was calculated, and the results are shown in Table 4 below.

As shown in Table 4 above, compared to Comparative Examples 1 to 3, which were single treatment groups of DMBQ, 2-MBQ, and Lapachone, respectively, when apigenin and quercetin were included in the quinone, either individually or in combination, a higher SIRT1 expression level was observed.

The composition containing all of quinone, apigenin, and quercetin showed the highest SIRT1 expression level.

Therefore, it was confirmed that a composition including at least one selected from the group consisting of quinone, apigenin, and quercetin exhibited a higher SIRT1 expression level compared to a single treatment group of quinone, and thus it can be seen that it can exhibit excellent effects on neurodegenerative diseases, aging-related diseases, and hangovers.

[Experimental Example 3]

Evaluation of blood alcohol and acetaldehyde decomposition ability

The following experiment was conducted to evaluate the blood alcohol and acetaldehyde decomposition ability of the composition of the present invention.

Specifically, 6-week-old SD rats were acclimated to solid mixed feed and water for 1 week and then housed in each cage, 2 rats each. The animal enclosure was maintained at a temperature of 22±2℃ and a humidity of 55%, and a 12-hour light/dark cycle was maintained, and drinking water and feed were freely available. After 18 hours of fasting, 100 mg/kg of the composition of the present invention was administered, and 30 minutes later, 3 g/kg of 30% ethanol was administered, and then the rats' orbital vein blood samples were collected at regular intervals. The blood obtained through the blood collection was centrifuged at 4℃ and 3,000 rpm for 10 minutes, and then the plasma was separated. The blood alcohol concentration (g/L) was measured using an ethanol analysis kit (Roche Co. Ltd., Germany), and the blood acetaldehyde concentration was measured using an acetaldehyde analysis kit (megazyme, K-ACHYD).

The experimental results for blood alcohol concentration are shown in Table 5 and Figure 1 below, and the blood acetaldehyde concentration is shown in Table 6 and Figure 2 below. In Example 13 below, a composition was used in which “wheat germ extract” and “chamomile extract” were mixed in a ratio of 4:1.

As shown in Table 5 and Fig. 1, the blood alcohol concentration was measured, and it was confirmed that the blood alcohol concentration of the wheat germ extract administration group (Comparative Example 8) and the mixture administration group (Example 13) decreased compared to the ethanol administration group (Comparative Example 7). In particular, after 90 minutes, the wheat germ extract administration group (Comparative Example 8) and the chamomile extract administration group (Comparative Example 9) decreased by 0.5 g/L and 0.4 g/L, respectively, compared to the ethanol administration group (Comparative Example 7), whereas the mixture administration group (Example 13) decreased by approximately 1.1 g/L.

In addition, as shown in Table 6 and Fig. 2, the blood acetaldehyde concentration was measured, and it was confirmed that the blood acetaldehyde concentration of the wheat germ extract administration group (Comparative Example 8) and the mixture administration group (Example 13) decreased compared to the ethanol administration group (Comparative Example 7).

In particular, after 60 minutes, the wheat germ extract administration group (Comparative Example 8) and the chamomile extract administration group (Comparative Example 9) showed a decrease of 6.7 mg/L and 3.9 mg/L, respectively, compared to the ethanol administration group (Comparative Example 7), while the mixture administration group (Example 13) showed a decrease of approximately 10.7 g/L.

In addition, after 90 minutes, it was confirmed that the wheat germ extract administration group (Comparative Example 8) and the chamomile extract administration group (Comparative Example 9) decreased by 9.8 mg/L and 6.5 mg/L, respectively, compared to the ethanol administration group (Comparative Example 7), while the mixture administration group (Example 13) decreased by approximately 16.9 g/L.

In the ethanol administration group (Comparative Example 7), the blood acetaldehyde concentration steadily increased for up to 90 minutes, whereas in the mixture administration group (Example 13), the blood acetaldehyde concentration increased for up to 30 minutes and then continuously decreased thereafter, confirming that the mixture of wheat germ extract and chamomile extract rapidly reduced blood acetaldehyde.

Finally, from the above experimental results, it can be seen that a composition containing a wheat germ extract, particularly a composition additionally containing a chamomile extract, has excellent alcohol and acetaldehyde decomposition effects.

[Experimental Example 4]

Evaluation of alcohol decomposition ability through NAD+/NADH ratio analysis

In order to evaluate the alcohol decomposition ability of the composition of the present invention, it was confirmed through NAD+/NADH ratio analysis.

In alcohol metabolism, a smooth supply of NAD+ is more important than the enzymatic activity of alcohol dehydrogenase (ADH) in determining the rate of alcohol metabolism. Excessive accumulation of NADH inhibits various intermediate metabolic processes, causing indirect hepatotoxicity. Therefore, rapid oxidation of accumulated NADH to NAD can alleviate alcohol-induced liver toxicity. Therefore, an increased NAD+/NADH ratio indicates smooth alcohol metabolism.

Hepatoma cell line (HepG2) was purchased from ATCC (American Type Culture Collection). The cells were cultured in DMEM-F12 (Dulbecco's Modified Eagle's Medium) containing 10% FBS and 1% PS at 37°C and 5% _CO2 . The cultured cells were seeded at 1Х10 ⁵ cells per 24-well plate for experiments. Two hours after seeding, 100 ug/ml of the compositions in Table 7 below were treated, and 24 hours later, the cells were used for NAD ⁺ /NADH analysis (Picoscan, BM-NDH-100, BioMax).

(In Example 14 below, a composition was used in which wheat germ extract, chamomile extract, and pine tree extract were mixed in a ratio of 4:1:1.)

The cells were obtained, washed with PBS, and centrifuged. NAD/NADH Extraction Buffer was added to the cell pellet obtained by centrifugation. The freezing and thawing were repeated twice to lyse the cells. Then, centrifugation (14,000 g, 5 min) was performed to obtain the supernatant. This supernatant was used as NADt. To prepare the NADH sample, NADt was heated, and then the supernatant from centrifugation (14,000 g, 1 min) was used as the NADH sample. For colorimetric analysis, the concentration of NADH was obtained according to the kit manual, and the NAD ⁺ /NADH ratio was calculated by calculating the ratio using the following formula.

● Concentration of NADH in the sample (C) = B / V x D (pmol/μl)

B: Amount of NADH in the measurement well (pmol)

V: Volume of sample dispensed into the well (μl)

D: Sample dilution ratio

● NAD/NADH ratio = (NADt - NADH) / NADH

The results of the above experiment are shown in Figure 3. As shown in Figure 3, the NAD+/NADH ratio increased in the wheat germ extract treatment group and the mixture treatment group compared to the control group, with the increase being significantly the greatest in the mixture treatment group. Therefore, it can be seen that wheat germ extract and the mixture have excellent hangover prevention or improvement effects.

In addition, after treating the samples, cells were disrupted, proteins were extracted, and sirt1 protein expression was confirmed through Western blotting, and the results are shown in Figure 4. As shown in Figure 4, the Sirt1 ratio was confirmed to have increased in the wheat germ extract and mixture treatment groups compared to the control group. In particular, the mixture treatment group showed the greatest increase, demonstrating a significantly superior effect.

[Experimental Example 5]

Evaluation of anti-neuroinflammatory activity through NO production analysis

In order to evaluate the anti-neuroinflammatory activity of the composition of the present invention, the composition of the present invention was treated on a neuronal cell line (BV2) and the change in NO production was analyzed.

Specifically, BV2 cells (microglia) were seeded in DMEM culture medium at a concentration of 6 Х10⁴cells/well in a 96-well plate, treated with 100 ug/ml of the composition in Table 8 below, and cultured for 24 hours. During this time, they were stimulated with 1 μg/ml of LPS. The production of nitrite, a soluble oxidation product of nitric oxide (NO), was measured in the culture medium through the Griess reaction, and the results are shown in Fig. 5.

As shown in Fig. 5, it was confirmed that when treated with the wheat germ extract and mixture of the present invention, NO production was reduced compared to the control group, and in particular, when treated with the mixture, NO production was significantly reduced the most. Therefore, it can be seen that the wheat germ extract and mixture of the present invention have excellent anti-neuroinflammatory effects.

Claims (21)

(1) Quinone or wheat germ extract; and (2) A composition comprising at least one selected from the group consisting of apigenin or chamomile extract, and (3) quercetin or chamomile extract. A composition comprising (1) quinone or wheat germ extract and (2) apigenin or chamomile extract in claim 1. In claim 1, a composition comprising (1) quinone or wheat germ extract and (3) quercetin or syringa extract. A composition comprising (1) quinone or wheat germ extract; (2) apigenin or chamomile extract; and (3) quercetin or chamomile extract in claim 1. A composition comprising wheat germ extract, chamomile extract and pine tree extract in claim 1. A composition comprising quinone, apigenin and quercetin in claim 1. A composition in claim 1, wherein the quinone is at least one selected from the group consisting of 2,6-DMBQ (2,6-dimethoxy-1,4-benzoquinone), 2-MBQ (2-methoxy-1,4-benzoquinone), and rapachone. A composition in claim 1, wherein the wheat germ extract, chamomile extract and pine tree extract are extracted with a solvent selected from the group consisting of water, C1 to C6 alcohol and mixed solvents thereof. A composition in claim 1, wherein the wheat germ extract, chamomile extract and pine tree extract are 0.01% to 99% ethanol extracts. In claim 12, the composition wherein the wheat germ extract, chamomile extract and pine tree extract are 50% to 70% ethanol extracts. A composition in claim 1, wherein the wheat germ extract, chamomile extract and pine tree extract are 60% ethanol extracts. In the first paragraph, the composition is a composition obtained by extracting a mixture of two or more selected from the group consisting of wheat germ, chamomile, and cinnamon tree, or by mixing extracts of each of these. A composition according to any one of claims 1 to 12, wherein the composition increases SIRT1 gene activity or SIRT1 gene expression. A pharmaceutical composition according to any one of claims 1 to 12, wherein the composition prevents or treats an aging-related disease. A pharmaceutical composition according to claim 14, wherein the age-related disease is at least one selected from the group consisting of geriatric cardiovascular disease and disorder, geriatric lung disease, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, osteoarthritis, geriatric ocular disease, and skin aging. A pharmaceutical composition according to any one of claims 1 to 12, wherein the composition prevents or treats a neurodegenerative disease. In claim 16, the neurodegenerative disease is at least one selected from the group consisting of stroke, cerebral infarction, cerebral embolism, ischemic neurological disease, degenerative neurological disease, apoplexy, Alzheimer's disease, Parkinson's disease, dementia, Huntington's disease, Pick's disease, Creutzfeldt-Jakob disease, multiple sclerosis, ischemic encephalopathy, epilepsy, Lou Gehrig's disease, memory loss, traumatic or disease-induced nerve cell damage, concussion, progressive supranuclear palsy, multiple system atrophy, olivary-pontocerebellar atrophy (OPCA), Shy-Drager syndrome, striatonigral degeneration, amyotrophic lateral sclerosis (ALS), essential tremor, corticobasal degeneration, diffuse Lewy body disease, Parkinson's-ALS-dementia complex, and Friedreich's ataxia. A pharmaceutical composition according to any one of claims 1 to 12, wherein the composition prevents or treats a hangover. A pharmaceutical composition according to claim 18, wherein the composition decomposes alcohol or acetaldehyde in blood. A food composition according to any one of claims 1 to 12, wherein the composition prevents or improves an aging-related disease, a neurodegenerative disease, or a hangover. An animal feed composition according to any one of claims 1 to 12, wherein the composition prevents or improves aging-related diseases, neurodegenerative diseases, or hangovers.