WO2017030167A1 - Agent for reconstructing/activating neural network - Google Patents

Abstract

Provided is an agent that is effective, inter alia, for improving memory and cognitive function. The agent is composed of: at least one component (A) selected from naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof, and salts thereof; and a Siberian ginseng component (B).

Description

Neural network reconstruction / activator

The present invention relates to agents, compositions, and foods useful for the treatment of neurological diseases, improvement of memory ability, extension of nerve axons, and the like.

Naringenin is a component contained in grapefruit and the like, and is considered to be a component having an antioxidant effect and the like.

In recent years, it has been reported that naringenin is effective in the treatment of Alzheimer's disease (Non-Patent Document 1), but its mechanism and action mechanism have not been clarified.

In addition, the herbal medicine bone crush (Katsusaiho) is a rhizome of the sea urchinaceae Hakamauraboshi, and is known to contain naringenin, but the traditional effects of bone crushing are as follows: Although anti-inflammation, muscular bone pain, tinnitus, toothache and the like are known (for example, Patent Document 1), the relationship between osteoclastic prosthesis and its active ingredient and the neural circuit is not known at all.

On the other hand, Ezokogi is a plant belonging to the family Uleaceae that grows naturally in Hokkaido, etc., and is considered to have a tonic effect.

JP 2005-330290 A

An object of the present invention is to provide a novel agent, composition, and food and drink useful for prevention / treatment of neurological diseases, improvement / improvement of memory ability and cognitive function, and the like.

神 経 Neurodegenerative diseases such as Alzheimer's disease, senile dementia, cerebrovascular dementia, and Parkinson's disease all have different etiologies, but all indicate symptoms that impair memory and cognition due to the breakdown of the neural network. For these diseases for which there is no effective therapeutic method, there is a real need for a treatment that can bring the nerve function closer to normal even when the nerve network has already been impaired. Moreover, considering the patient's standpoint, treatment with less burden is more desirable than nerve cell transplantation and gene therapy that require surgery. Therefore, it is desired to develop a drug that regenerates a neural network even when a nerve cell is damaged or after it has been damaged.

Also, amyotrophic lateral sclerosis is an intractable disease in which the limbs of the cerebral cortex motor cortex and the motor neurons of the spinal cord and brainstem drop off, and there is no effective treatment. Furthermore, motor neurons in the cerebral cortex motor cortex are also damaged by cerebral hemorrhage, cerebral infarction, brain tumor, brain trauma, etc., leading to paralysis of the body. Traumatic spinal cord injury also results in limb paralysis due to damage to spinal motor neurons. In addition, multiple cerebral infarction is a demyelinating disease of the central nervous system caused by abnormal activation of autoimmunity, but axonal loss occurs as the disease progresses, which is an irreversible malfunction of the neural network. Leads to. In either case, it is a dysfunction due to the breakdown of the neural network, but if the surviving nerve cells are activated to form a neural network again, recovery of the function can be expected.

In view of the circumstances as described above, the present inventor asked herbal medicines for a component exhibiting excellent activity for improving the atrophy / degeneration of nerve axons in the brain that has become degenerated and dysfunctional. We have been conducting research based on the idea that by exploring the introduction, we can find new compositions that could not be expected from the pathological analysis of conventional neurodegenerative diseases.

As a result, specific ingredients such as naringenin [and herbal medicine ingredients containing such specific ingredients (such as osteoclastic components)] and sorghum ingredients are effective in repairing neurites and improving memory impairment. I found out. Such functions also lead to the prevention and treatment of neurological diseases. Therefore, the specific component and the sorghum component (particularly a combination of these) are used for the prevention and treatment of neurological diseases, memory ability. The present inventors have obtained knowledge that it is effective for use in improving cognitive function and for repairing and extending neurites, and have further studied based on this knowledge to complete the present invention.

That is, the agent of the present invention comprises at least one component (A) selected from naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof, and salts thereof. Contains Ezokogi component (B).

Such an agent of the present invention may be an agent for use in at least one application selected from the following (1), (2) and (3).
(1) Neurite repair and / or extension (growth)
(2) Improvement or improvement of memory and / or cognitive function (3) Prevention and / or treatment of neurological diseases

It should be noted that such an agent of the present invention usually contains only naringenin as an active ingredient and often does not contain an agent for the prevention and / or treatment of Alzheimer's disease.

In the agent of the present invention, the component (A) may contain, for example, at least one component selected from naringenin, a glycoside of naringenin (for example, naringin, naringen glucuronide), and salts thereof.

In addition, the agent of the present invention may contain an osteoclastic component (such as an osteoclastic extract) as a component containing the component (A).

The agent of the present invention may be used in combination of the component (A) and the component (B), and particularly may contain the component (A) and the component (B).

In such a combination aspect, the agent of the present invention contains a bone crush extract as the component (A) and a carp extract as the component (B), and the ratio of the bone crush extract and the carp extract is the former / The latter (weight ratio) = 1 / 0.01 to about 1/100 may be used.

In addition, the agent of the present invention may further combine at least one component (C) selected from diosgenins (such as diosgenin and diosine), dihydroxyvitamin D3, and desonomin. In such a combination mode, the component (C) may be a different agent from the component (A) and the component (B), or may be an agent (mixture) containing the component (A) and the component (B). .

In such an agent, the ratio of the component (A) and / or the component (B) to the component (C) may be about the former / the latter (weight ratio) = 1 / 0.01 to 1/100. .

The present invention includes at least one component (A) selected from naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof, and salts thereof, and an elephant component A composition containing (B) is also included. Such a composition may be a pharmaceutical composition.

In such a composition, the component (A) may contain, for example, at least one or more components selected from naringenin, glycosides of naringenin (eg, naringin, naringen glucuronide), and salts thereof. .

In addition, the composition of the present invention may contain an osteoclastic component (such as an osteoclastic extract) as the component containing component (A). A typical composition of the present invention contains a bone crush extract as the component (A) and a carp extract as the component (B), and the ratio between the bone crush extract and the carp extract is the former / the latter ( (Weight ratio) = 1 / 0.01 to 1/100.

Moreover, the composition of the present invention may further combine at least one component (C) selected from diosgenins (such as diosgenin and diosine), dihydroxyvitamin D3, and desonomin. In such a combination mode, the component (C) may be a composition different from the component (A) and the component (B), or may be a composition containing the component (A) and the component (B).
In such a composition, the ratio of the component (A) and / or the component (B) to the component (C) may be about the former / the latter (weight ratio) = 1 / 0.01 to 1/100. Good.

In particular, the composition of the present invention may be a composition for use in at least one application selected from the following (1), (2) and (3).
(1) Neurite repair and / or extension (growth)
(2) Improvement or improvement of memory and / or cognitive function (3) Prevention and / or treatment of neurological diseases

Included in the present invention are foods and drinks containing the above-mentioned agent (particularly, an agent in which component (A) and component (B) are combined) or the composition of the present invention.

Such foods and drinks may be supplements, health foods, functional indication foods, nutritional functional foods, or foods for specified health use.

In addition, the food or drink of the present invention may be a food or drink containing 1 to 90% by weight of the osteoclast extract, or a food or drink containing 1 to 90% by weight of the elephant extract.

A typical food and drink product of the present invention contains 1 to 90% by weight of a bone crush extract and 1 to 90% by weight of an elephant extract, and the ratio of the osteocrush extract to the elephant extract is the former / the latter (weight ratio). ) = 1 / 0.01 to 1/100.

Such a food or drink may contain at least one component (C) selected from diosgenins (such as diosgenin and diosin), dihydroxyvitamin D3, and desonomin. In such a food or drink, the proportion of component (C) may be, for example, about 1 to 90% by weight.
In such a food or drink, the ratio of component (A) and / or component (B) to component (C) is about the former / the latter (weight ratio) = 1 / 0.01 to 1/100. May be.

According to the present invention, the neural network can be reconstructed and / or activated. Such reconstruction and activation are usually accompanied by repair and / or extension (extension) of neurites (axons, dendrites, etc.). Therefore, the agent or composition of the present invention is useful for preventing, treating or improving various neurological diseases caused by degeneration or atrophy of axons and dendrites, as well as improvement or improvement (enhancement) of the cognitive memory function. It is valid.

Moreover, since the improvement of the memory ability in the cognitive function is recognized in the agent and composition of the present invention, regardless of whether or not it leads to a neurological disease, the ingredients of the food and drink for the purpose of improving and improving the memory ability, etc. It is also useful.

FIG. 1 is a diagram showing an axonal extension action by a bone crush extract and a bone crush component on normal cerebral cortical neurons. FIG. 2 is a diagram showing an axonal re-extension effect when a crushed prosthetic component is later treated against Aβ (25-35) -induced axonal atrophy in cerebral cortical neurons. FIG. 3 is a diagram showing the area of amyloid plaques in the prefrontal cortex of 5XFAD mice administered with a bone fracture supplement for 31 days. FIG. 4 is a diagram showing the area of the degenerated axon terminal area in the prefrontal cortex of 5XFAD mice administered with the osteoclastic prosthesis for 31 days. FIG. 5 is a diagram showing the area of amyloid plaques in the peri-olfactory cortex of 5XFAD mice to which the osteoclastic extract was administered for 31 days. FIG. 6 is a diagram showing the area of the degenerated axon terminal area in the perianptic cortex of 5XFAD mice administered with the osteoclastic extract for 31 days. FIG. 7 is a diagram showing the area of amyloid plaques in the hippocampal CA1 region of 5XFAD mice administered with the osteoclastic prosthesis for 31 days. FIG. 8 is a diagram showing the area of the degenerated axon terminal area in the hippocampal CA1 region of 5XFAD mice administered with the osteoclastic prosthesis for 31 days. FIG. 9 is a diagram showing the area of amyloid plaques in the hippocampal dentate gyrus region of 5XFAD mice to which the osteoclastic extract was administered for 31 days. FIG. 10 is a diagram showing the area of the degenerated axon terminal area in the hippocampal dentate gyrus region of 5XFAD mice administered with the osteoclastic prosthesis for 31 days. FIG. 11 is a diagram showing the level of phosphorylation of threonine of the 514th amino acid of CRMP2 in the cerebral cortex of 5XFAD mice administered with the osteoclastic extract for 22 days. FIG. 12 is a diagram showing an axonal re-extension effect when a fraction of the osteoclastic extract is treated later against Aβ (25-35) -induced axonal atrophy in cerebral cortical neurons. FIG. 13 is a diagram showing an LC-MS chart of the n-butanol fraction of the osteoclastic extract. FIG. 14 is a diagram showing the axonal re-extension effect when a compound contained in the osteoclastic extract is later treated against Aβ (25-35) -induced axonal atrophy in cerebral cortical neurons. FIG. 15 is a diagram showing the results of an object recognition memory test in 5XFAD mice administered with a bone crush extract, or an extract of Ekokogi leaves, or a combination of an osteoclastic extract and an extract of Ekokogi leaves for 15 days. FIG. 16 is a diagram showing the results of an object recognition memory test in normal mice administered with Ezocogi leaf extract for 27 days.

<Agent / Composition>
The agent or composition of the present invention comprises a specific component (A) or an sorghum component (B).

Such an agent or composition (and food or drink) of the present invention is useful for the reconstruction and / or activation of a neural network.

Specifically, the agent or composition of the present invention often has a function of repairing or extending (extending) neurites [particularly atrophy or deficient neurites (axons or dendrites)].
In addition, the agent or composition of the present invention is often associated with such a function, or has an effect of improving or improving memory ability and / or cognitive function, and an effect of preventing and / or treating a neurological disease. .

Therefore, the agent or composition of the present invention may be suitably used for at least one application selected from the following (1), (2), and (3).
(1) Neurite repair and / or extension (growth)
(2) Improvement or improvement of memory ability and / or cognitive function [especially memory ability and / or cognitive function reduced due to atrophy or loss of neurites (axons or dendrites)] (3) Neurological diseases [especially nerves And / or treatment of neurological diseases associated with atrophy or loss of processes (axons and dendrites)

In the above (3), the neurological disease may be a traumatic neurological disease or a neurodegenerative disease. For example, Alzheimer's disease, dementia (non-Alzheimer type dementia such as senile dementia, Cerebrovascular dementia, Pick's disease, frontotemporal dementia, Lewy body dementia, vascular dementia, etc.), Parkinson's disease, Huntington's disease, brain contusion, spinal cord injury, amyotrophic lateral sclerosis, multiple occurrences Include cerebral infarction.

Of these, the agent or composition of the present invention may prevent or treat one or more neurological diseases.

[Component (A)]
Component (A) consists of naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof [or glycosides (O-glycosides)], and salts thereof (especially pharmaceutically acceptable) Selected salt).

The sugar constituting the glycoside is not particularly limited, and is a monosaccharide [eg, glucose, galactose, apiose, rhamnose, xylose, uronic acid (glucuronic acid, etc.)], disaccharide (eg, rutinose, neohesperidose, etc.) , Gentiobiose, etc.), and sugars to which these sugars are bonded.

The glycoside may be bound with one or more sugars.

Specific glycosides include, for example, naringenin glycoside [eg, naringenin glucoside (eg, naringenin-7-glucoside, naringenin-4-glucoside, etc.), naringening glucuronide (eg, naringenin-4-glucuronide, naringenin- 4-glucuronide etc.), naringenin rutinosides (eg naringin etc.)], eriodictyol glycosides [eg eriodictyol rutinosides (eg neoeriocitrin etc.)], 5,7-dihydroxychromone Glucose [eg, 5,7-dihydroxychromone neohesperidoside (eg, 5,7-dihydroxychromone-7-neohesperidoside, etc.)], caffeic acid glycoside [eg, caffeic acid glucoside (eg, Caffeic acid-4-glu Sid, etc.), etc.], protocatechuic acid glycosides [e.g., protocatechuic acid glucoside (e.g., such as protocatechuic acid 3-glucoside), etc.] and the like.

The salt is not particularly limited, and examples thereof include metal salts [for example, alkali metal salts (for example, lithium salt, sodium salt, potassium salt), alkaline earth metal salts (for example, magnesium salt, calcium salt), periodic table, etc. Group 13 metal salts (for example, aluminum salts), transition metal salts (for example, salts such as zinc), etc.] ammonium salts, amine salts [for example, alkylamine salts (for example, trialkylamine salts, trialkylamine salts of triethylamine salts) ), Alkanolamine salts (for example, monoethanolamine salts, triethanolamine salts), cyclic amine salts (for example, pyridine salts)] and the like.

Component (A) may contain one or more components. When component (A) contains two or more components, it may contain two or more components that are common in the basic skeleton (for example, including naringenin and naringenin glycoside), and combinations of components having different basic skeletons 2 or more may be included (for example, having a combination of naringenin or its glycoside and eriodictyol or its glycoside).

Among these, component (A) is typically at least one component selected from naringenin, a glycoside of naringenin (naringin, naringen glucuronide, etc.), and salts thereof (collectively, naringenins) May be included) at least.

The component (A) may be synthesized or derived from a natural product. As a synthesis method, a conventional method can be used.

For example, Naringenin and Naringin are known to be contained in citrus plants (citrus plants such as grapefruit and hassaku), and the component (A) may be obtained from such plants.

In particular, osteoclasts often contain other components (A) in addition to naringenin, etc., and in combination with these, the effects of the present invention are effectively expressed. Cheap.

Therefore, the agent of the present invention may contain a bone crush prosthetic component as a component containing the component (A).

Examples of the osteoclastic component include osteoclastic prosthesis, osteoclastic extract (osteoclast extract) and the like, and these may be combined.

A commercially available product may be used as the osteoclastic extract, and the osteoclastic extract is extracted by a conventional method {for example, using an extraction solvent [for example, water, alcohol (methanol, ethanol, etc.), a mixture thereof, etc.]]. It can also be obtained by subjecting to extraction at ordinary temperature or under heating by a conventional method or a method according thereto.

Among the extracts, in particular, a part having a high content or concentration of component (A) may be separated or fractionated. For example, an aqueous solvent extract of osteoclast (for example, an extract using water, a mixed solvent of water and alcohol as an extraction solvent), an alcohol [for example, n-butanol and the like having 3 or more carbon atoms (for example, n-butanol) Processed products obtained by separation or fractionation by extraction or the like with an alcohol having about 4 to 6 carbon atoms] often contain a high content of component (A) containing naringenin. You may use a processed material suitably.

In the extraction, the osteoclastic prosthesis may be used as it is or after being crushed (crushed). The extract may be further subjected to concentration and drying (room temperature drying, freeze drying, etc.).

The form of the osteoclastic component is not particularly limited, but may be, for example, powder (or powder).

[Ezokogi component (B)]
Examples of the Ekokogi component (B) include Ezoukogi, Ezoukogi extract (Ezoukogi extract), and the like.

Ezoukogi [scientific name: Eleutherococcus senticosus, Acanthopanax senticosus (Rupr. May be used, these may be used in combination, or the whole plant may be used.

In particular, at least the leaves of Ezokogi may be suitably used.

The Ekokogi extract may be a commercially available product or can be obtained by extracting Ezokogi by a conventional method.

Examples of the extraction solvent include water, alcohol [eg, alkanol (eg, C _1-4 alkanol such as methanol, ethanol, isopropanol, n-butanol), polyol (eg, ethylene glycol, propylene glycol, 1,3-butylene]. Glycol, glycerin, etc.), ketones (eg, acetone, methyl ethyl ketone, etc.), ethers (eg, diethyl ether, etc.), esters (eg, ethyl acetate, etc.) and the like.
You may use an extraction solvent individually or in combination of 2 or more types.
Typical extraction solvents include water, alcohol (such as ethanol), a mixed solution thereof, and the like.

Extraction temperature and extraction time are not particularly limited and can be appropriately selected according to the extraction solvent, extraction method, and the like.

In the extraction, the elephant may be subjected to an appropriate treatment (pulverization treatment, heat treatment, drying treatment, etc.) as necessary. The extract may be further subjected to concentration and drying (room temperature drying, freeze drying, etc.).

The form of the Ekokogi component is not particularly limited, but may be powdery (or powdery).

[Component (C)]
In the agent or composition of the present invention, component (A) and / or component (B) may be combined with diosgenins, dihydroxyvitamin D3, desonomin and the like. By combining with these components (hereinafter, these may be referred to as component (C)), the effects of the present invention may be realized more efficiently.

Among these, in particular, diosunigens may be preferably used. Therefore, component (C) may be composed of at least diosgenins.
Examples of diosgenins include diosgenin, diosgenin derivatives, and salts thereof.

As the diosgenin derivative, a compound in which the hydroxyl group at the C3 position of diosgenin is substituted, a compound having a substituent at the C2 position (or a compound having a hydrogen atom at the C2 position substituted), a compound having a substituent at the C4 position (or C4) Compound having a hydrogen atom at the position), a compound having a substituent at the C6 position (or a compound having the hydrogen atom at the C6 position substituted) [for example, ester derivatives of the hydroxyl group at the C3 position (for example, amino acid derivatives, aminosulfones) Acid derivatives, carbamate derivatives), halogenated derivatives of the hydroxyl group at the C3 position, etc.], and glycosides thereof.

Specific examples of the diosgenin derivative include a compound represented by the following formula (I-1), a glycoside of diosgenin or a compound represented by the following formula (I-1), and the like.

(In the formula, R ¹ , R ² , R ³ and R ⁴ are the same or different and represent a hydrogen atom or a substituent. However, when R ² , R ³ and R ⁴ are hydrogen atoms, R ¹ is hydroxyl. Not a group.)

In R ¹ , the substituent includes a hydrocarbon group {eg, alkyl group [eg, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group]. Group, linear or branched alkyl group such as pentyl group (eg C _1-12 alkyl group, preferably C _1-8 alkyl group)], cycloalkyl group (eg cyclopentyl group, cyclohexyl group, cycloheptyl) Group, a C _4-10 cycloalkyl group such as a cyclooctyl group, preferably a C _5-8 cycloalkyl group), an aralkyl group (for example, a C _6-10 aryl C _1-4 alkyl group such as a benzyl group or a phenethyl group) Saturated with a polycyclic aliphatic hydrocarbon group (for example, a decalinyl group, a norbornyl group, an adamantyl group, a dimethyladamantyl group, etc.) Saturated aliphatic hydrocarbon group; an aryl group (e.g., phenyl group, a tolyl group, C _6-10 aryl group such as xylyl) aromatic hydrocarbon group}, a hetero atom (a nitrogen atom such as an oxygen atom, a sulfur atom, Phosphorus atom etc.) containing group {eg oxygen atom containing group [eg hydroxyl group, oxo group (═O), group —OR ^a , group —O—CO—R ^a , group —O—CO—N (R ^b ) ₂ , group —O—CO—O—R ^a , group —O—CO—S—R ^a , group —OR ^c , group —O—SO ₂ —OH, group —O—PO ₂ —OH, group — (OR ^d ) _k —R ^e , carboxyl group, group —CO—O—R ^a , group —CO—N (R ^b ) ₂ etc.], nitrogen atom-containing group [for example, amino group, group —NR ^a R ^b A group —NR ^b —CO—O—R ^a , a group —NR ^b —CO—N (R ^b ) ₂ , a nitrogen-containing ring group (eg Group corresponding to pyridine, pyrroline, pyrrole, indole, etc.)], a sulfur atom-containing group [eg, mercapto group, group —SR ^a , group —S—S—R ^a , sulfo group (—SO ₃ H) , Group —SO ₂ —R ^b , group —S—CO—N (R ^b ) _{2 and the} like], phosphorus atom-containing group [eg, phosphate group (H ₂ PO ₄ —), group —PO ₃ H and the like], An amino acid group [or a residue of an amino acid, for example, formed by an ester bond between a hydroxyl group at position 3 constituting diosgenin (corresponding to the substitution position of R ^{1 in} the above formula) and a carboxyl group of an amino acid (glycine, alanine, etc.) Group], a halogen atom (for example, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, etc.).

In the above formula, R ^a is a hydrocarbon group (eg, the exemplified hydrocarbon group such as an alkyl group), and R ^b is a hydrogen atom or a hydrocarbon group (eg, the exemplified hydrocarbon group such as an alkyl group). ), R ^c is a sugar (or sugar chain or sugar residue), R ^d is an alkylene group (eg, C _2-4 alkylene group such as ethylene group, propylene group, trimethylene group, etc.), R ^e is a hydrogen atom, hydroxyl group A group or a hydrocarbon group (for example, the above exemplified hydrocarbon group such as an alkyl group (such as a methyl group)), k represents an integer of 2 or more (for example, 2 to 10), and R ^a and R ^b are the same Or they may be different groups, and when R ^b is plural, they may be the same or different.

In R ^a and R ^b , the hydrocarbon group (such as an alkyl group) may further have a substituent. The substituent is not particularly limited, but includes the above-exemplified substituents, for example, an oxygen atom-containing group (for example, a hydroxyl group, a carboxyl group, a group —OR ^a , a group —O—CO—R ^a, etc.), a nitrogen atom-containing group A group (for example, an amino group, a group —NR ^a R ^b ), a sulfur atom-containing group (for example, a mercapto group, a group —SR ^a , a sulfo group, a group —SO ₂ —R ^b, etc.).
The hydrocarbon group may have these substituents alone or in combination of two or more.
When the hydrocarbon group has a substituent, the number of substituents may be 1 or more, for example, 1 to 10 (eg, 1 to 8), preferably 1 to 6 (eg, 1 to 4), Preferably, it may be about 1 to 3.

When R ¹ is a substituent, representative R ¹ includes, for example, a hydrocarbon group [eg, alkyl group (eg, group — (CH ₂ ) _n —CH ₃ ), cycloalkyl group, aralkyl group, etc.] , Heteroatom-containing groups {eg oxygen atom-containing groups [eg hydroxyl groups, groups —O— (CH ₂ ) _n —CH ₃ , groups —O— (CH ₂ ) _m —NH ₂ , groups —O— (CH ₂ ) _m —COOH, group —O— (CH ₂ ) _m —SO ₃ H, group —O—CO— (CH ₂ ) _n —CH ₃ , group —O—CO—NH— (CH ₂ ) _n —CH ₃ , group —O—CO—NR— (CH ₂ ) _n —CH ₃ , group —O—CO—NH—CH (R ^b ) —COOH, group —O— (CH ₂ ) _n —CO—NH—AD (In the formula, AD is an adamantyl group (1-adamantyl group, 2,6-dimethyladamantan-1-yl A group, etc.)), groups _{-O-CO-NH- (CH 2} ) m -SO 3 H, group _{-O-CO-NH- (CH 2} ) m -COOH, group -O-CO-O- ( CH ₂ ) _n —CH ₃ , group —O—CO—S— (CH ₂ ) _n —CH ₃ , group —O—SU (wherein SU represents a sugar chain), group —O—SO ₂ — OH, group —O—PO ₂ —OH, group — (OCH ₂ CH ₂ ) _m —CH ₃ , group — (OCH ₂ CH ₂ CH ₂ ) _m —CH ₃ , carboxyl group, group —COO (CH ₂ ) _n CH ₃ , group —CO—NH— (CH ₂ ) _n —CH ₃ , group —SO ₃ H, group —SO ₂ — (CH ₂ ) _n —CH ₃ , group —SO ₂ —Ph where Ph is A phenyl group)), a group —CO—NH—CH (R ^b ) —COOH, a group —CO—NH— (CH ₂ ) _n —SO ₃ H, etc.], A nitrogen atom-containing group [for example, an amino group, a group —NH— (CH ₂ ) _n —CH ₃ , a group —NH— (CH ₂ ) _n —NH _{2, a} group —NH—CH (R ^b ) —COOH, a group — NH— (CH ₂ ) _m —SO ₃ H, group —NH— (CH ₂ ) _m —SO ₂ H, group —NH—CO—O— (CH ₂ ) _n —CH ₃ , group —NH—CO—NH ₂ , a group —NH—CO—NH—AD (wherein AD represents an adamantyl group (1-adamantyl group, 2,6-dimethyladamantan-1-yl group, etc.)), a group —NH—CO—NH— CH (R ^b ) —COOH, groups —NH—CO—NH— (CH ₂ ) _m —SO ₃ H, groups —NH—CO—NH— (CH ₂ ) _m —COOH, etc.], sulfur atom-containing groups [eg , Mercapto group, group -S- (CH ₂ ) _n -CH ₃ , group -S- (CH ₂ ) _m -C OOH, group —S— (CH ₂ ) _m —CH (NH ₂ ) —COOH, group —S—CO—NH—AD (where AD is an adamantyl group (1-adamantyl group, 2,6-dimethyladamantane— 1-yl group, etc.)), groups —SS— (CH ₂ ) _m —CH (NH ₂ ) —COOH, groups —SO ₃ H etc.], phosphorus atom-containing groups [eg group —PO ₃ H Etc.], amino acid groups (eg, group —O—CO—CH ₂ —NH ₂ etc.)}, halogen atoms (eg, fluorine atom, chlorine atom, bromine atom, iodine atom etc.) and the like.

In the above formula, m is an integer of 1 or more (eg, 1 to 10, preferably 1 to 4, more preferably 1 or 2), and n is an integer of 0 or more (eg, 0 to 10, preferably 0 to 2). 7) and R ^b is the same as the above [that is, a hydrogen atom or a hydrocarbon group (for example, an alkyl group, etc.)].

In R ² , R ³ and R ⁴ , examples of the substituent include the same substituents as those exemplified in the section of R ¹ .
When R ² and / or R ⁴ is a substituent, typical examples of the substituent include an oxygen atom-containing group, a nitrogen atom-containing group, a sulfur atom-containing group, an amino acid group, and a halogen atom.
In addition, when R ³ is a substituent, typical substituents include a halogen atom.

In the formula (I-1), combinations of R ¹ to R ⁴ are not limited, and all combinations are included. Typical combinations of R ¹ to R ⁴ include, for example, the following combinations.
(1) A combination in which R ¹ is a substituent other than a hydroxyl group, and R ² to R ⁴ are hydrogen atoms. (2) R ¹ is a substituent other than a hydroxyl group, R ² is a substituent, R ³ and R ⁴ are hydrogen. Combinations that are atoms (3) Combinations in which R ¹ is a substituent other than a hydroxyl group, R ³ is a substituent, R ² and R ⁴ are hydrogen atoms (4) R ¹ is a substituent other than a hydroxyl group, R ² and A combination in which R ³ is a substituent and R ⁴ is a hydrogen atom (5) A combination in which R ¹ is a substituent other than a hydroxyl group, and R ² , R ³ and R ⁴ are substituents (6) R ¹ is other than a hydroxyl group A combination in which R ² and R ³ are hydrogen atoms and R ⁴ is a substituent (7) R ¹ is a substituent other than a hydroxyl group, R ³ is a hydrogen atom, and R ² and R ⁴ are substituents Combination (8) R ¹ is hydroxyl A substituent other than a group, a combination in which R ² is a hydrogen atom, R ³ and R ⁴ are substituents (9) a combination in which R ¹ is a hydroxyl group, R ² is a substituent, and R ³ and R ⁴ are hydrogen atoms ( 10) A combination in which R ¹ is a hydroxyl group, R ³ is a substituent, R ² and R ⁴ are hydrogen atoms (11) R ¹ is a hydroxyl group, R ² and R ³ are substituents, and R ⁴ is a hydrogen atom Combination (12) A combination in which R ¹ is a hydroxyl group and R ² to R ⁴ are substituents (13) A combination in which R ¹ is a hydroxyl group, R ² and R ³ are hydrogen atoms, and R ⁴ is a substituent (14) A combination in which R ¹ is a hydroxyl group, R ² is a hydrogen atom, R ³ and R ⁴ are substituents (15) A combination in which R ¹ is a hydroxyl group, R ³ is a hydrogen atom, and R ² and R ⁴ are substituents

Representative diosgenin derivatives include, for example, the following formula (II)

(3β, 25R) -3- (2-aminoethanoyloxy) -spirost-5-ene {(3β, 25R) -3- (2-Aminoethanoyloxy) -spirost-5-ene}, represented by the following chemical formula ( III)

(3β, 25R) -3-fluorospirost-ene {(3β, 25R) -3-Fluorospirost-5-ene} represented by the formula (3β, 25R) -3- (2-aminoethylsulfonyloxy) -Spirost-5-ene {(3β, 25R) -3- (2-Aminoethylsulfonyl-oxy) -spirost-5-ene}, (3β, 25R) -3- (2-aminopropylsulfonyloxy) -spirost-5 -Ene, (3β, 25R) -3- [N- (2,6-dimethyladamantan-1-yl) carbamoyloxy] -spirost-5-ene, (3β, 25R) -3-{[N- (2 , 6-Dimethyladamantan-1-yl) carbamoyl] amino} -spirost-5-ene, (3β, 25R) -3- [N- (2,6-dimethyladamantan-1-yl) carbamoylthio] -spirost- 5-ene, (3β, 25R) -3-{[N- Adamantan-1-yl) carbamoyl] amino} -spirost-5-ene, (3β, 25R) -3- [N- (adamantan-1-yl) carbamoylthio] -spirost-5-ene, and (3β, 25R ) -3- [N- (adamantan-1-yl) carbamoyloxy] -spirost-5-ene [or (3β, 25R) -3- (1-adamantyl-aminocarbonyloxy) -spirost-5-ene {( 3β, 25R) -3- (1-adamantyl-aminocarbonyloxy) -spirost-5-ene}].

Examples of the glycoside include diosgenin or a compound in which R ¹ is a hydroxyl group in the above formula (I-1) and a sugar (such as the sugar exemplified above) is bonded to a hydroxy group (for example, diosine). .

Examples of salts (particularly pharmaceutically acceptable salts) include hydrohalides (for example, hydrofluorates, hydrochlorides, hydrobromides, hydroiodates, etc.) and inorganic acid salts. (Eg sulfate, nitrate, perchlorate, phosphate, carbonate, bicarbonate, etc.), organic carboxylates (eg acetate, oxalate, maleate, tartrate, fumarate, citric acid Acid salts), organic sulfonates (eg methanesulfonate, trifluoromethanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, camphorsulfonate, etc.), amino acid salts (eg aspartic acid) Salt, glutamate, etc.), organic amine salt (for example, trimethylamine, triethylamine, pyridine, picoline, dicyclohexylamine, N, N′-dibenzylethylenedi) Salts with organic bases such as min, arginine and lysine), quaternary amine salts, alkali metal salts (eg sodium salts, potassium salts etc.), alkaline earth metal salts (eg magnesium salts, calcium salts etc.), etc. .

Representative diosgenins include diosgenin, compounds represented by the above formula (I-1), diosin, and salts thereof. Particularly preferred diosgenins include diosgenin, a compound represented by the formula (II), a compound represented by the formula (III), diosine, and salts thereof.

[Form / Administration]
The agent or composition of this invention should just contain any one of a component (A) and a component (B), and may combine a component (A) and a component (B). Usually, in the composition of this invention, you may combine a component (A) and a component (B).
By combining the component (A) and the component (B), it is easier to obtain a memory improvement effect and the like.

When combining a component (A) and a component (B), a component (A) and a component (B) are good also as another agent or a composition, and the agent (A) containing a component (A) and a component (B) ( A mixture) or a composition may be used.

The same applies to the component (C).

When the component (A) and the component (B) are combined, these ratios can be appropriately selected according to the desired effect. For example, the ratio between the component (A) and the component (B) is the former / the latter (Weight ratio) = 1 / 0.001 to 1/1000, preferably 1 / 0.01 to 1/100, more preferably 1 / 0.02 to 1/50, particularly 1 / 0.1 to 1/10 It may be a degree.

In particular, in the case of combining the osteoclastic extract as the component (A) and the sorghum extract as the component (B), the ratio of the former / the latter (weight ratio) = 1 / 0.001 to 1/1000, Preferably, it may be 1 / 0.01 to 1/100, more preferably 1 / 0.02 to 1/50, and particularly about 1 / 0.1 to 1/10.

In addition, when the osteoclast supplement and the sorghum extract contain a solvent component etc., the said ratio is a ratio of an active ingredient (or solid content) conversion.

When the component (C) is combined, the ratio of the component (A) and / or the component (B) to the component (C) is the former / the latter (weight ratio) = 1 / 0.001 to 1/1000, preferably May be 1 / 0.01 to 1/100, more preferably 1 / 0.02 to 1/50, especially about 1 / 0.1 to 1/10.

The form of the agent or composition is not particularly limited, and component (A) and / or component (B) may be used as an agent (for example, powder) or composition as it is, and other components (carrier, excipient, etc.). You may formulate with it.

In particular, for formulation, for example, excipients, binders, disintegrants, coating agents, lubricants, colorants, flavoring agents, and if necessary, stabilizers, emulsifiers, absorption enhancers, surfactants, A pH adjusting agent, preservative, antioxidant and the like can be used, and it can be formulated by a conventional method by incorporating components generally used as a raw material of the formulation.

The component (carrier) used for the formulation is not particularly limited, but for example, animal and vegetable oils such as soybean oil, beef tallow and synthetic glycerides; hydrocarbons such as liquid paraffin, squalane and solid paraffin; octyldodecyl myristate, myristic acid Ester oil such as isopropyl; Higher alcohol such as cetostearyl alcohol and behenyl alcohol; Silicone resin; Silicone oil; Polyoxyethylene fatty acid ester, sorbitan fatty acid ester, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, poly Surfactants such as oxyethylene polyoxypropylene block copolymer; hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl polymer, polyethylene glycol Water-soluble polymers such as ethanol, polyvinylpyrrolidone and methylcellulose; lower alcohols such as ethanol and isopropanol; polyhydric alcohols such as glycerin, propylene glycol, dipropylene glycol and sorbitol; sugars such as glucose and sucrose; Inorganic powders such as aluminum magnesium silicate and aluminum silicate; purified water and the like.

Examples of excipients include lactose, corn starch, sucrose, glucose, mannitol, sorbitol, crystalline cellulose, silicon dioxide and the like.

Examples of the binder include polyvinyl alcohol, gelatin, methylcellulose, ethylcellulose, gum arabic, tragacanth, gelatin, shellac, hydroxypropylmethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, polyvinyl acetal diethylaminoacetate, corn starch and the like.

Examples of the disintegrant include corn starch, low-substituted hydroxypropylcellulose, crospovidone, crystalline cellulose, precipitated calcium carbonate, croscarmellose sodium, calcium citrate, dextrin, pectin, carboxymethylcellulose calcium and the like.

Examples of the lubricant include magnesium stearate, talc, polyethylene glycol, light anhydrous silicic acid, sucrose fatty acid ester, and the flavoring agents include cocoa powder, menthol, aroma powder, mint oil, dragonfly, For example, cinnamon powder is used.

It should be noted that the agent or composition of the present invention may be further combined with another component useful for improving neurological diseases and memory as long as the effects of the present invention are not impaired. As an aspect combined, the aspect used separately from an agent or a composition other than the aspect contained in an agent or a composition is mentioned.

Examples of the form (dosage form) of the agent or composition of the present invention include tablets, powders, fine granules, granules, dry syrups, coated tablets, orally disintegrating tablets, chewable tablets, capsules, soft capsules, and syrups. Agent, oral solution, troche, jelly, inhalant, suppository, injection, ointment, eye drop, eye ointment, nasal drop, ear drops, poultice, lotion, external solution, spray, External aerosols, creams, gels, tapes, buccal tablets, sublingual tablets, vaginal suppositories, vaginal tablets, rectal soft capsules and the like can be mentioned.

The administration (taking) form of the agent or composition of the present invention is not particularly limited, and may be oral administration or parenteral administration. Parenteral administration includes, for example, rectal administration, nasal administration, pulmonary administration, injection administration (eg, intravenous administration, intrathecal administration, intraepidural administration, intramuscular administration, subcutaneous administration, intraperitoneal administration) Intraarterial administration, intraarticular administration, intracardiac administration, intracapsular administration, intradermal administration, intralesional administration, intraocular administration, intrathoracic administration, subarachnoid administration, intrauterine administration, intraventricular administration) and the like.

A typical administration form is oral administration.

The amount of component (A) and / or component (B) contained in the agent or composition of the present invention is not particularly limited, and is preferably a dose sufficient to obtain a desired effect.
For example, the dose varies depending on, for example, the degree of symptom, age, sex, body weight, dosage form, salt type, specific type of disease, etc., but usually per day, component (A) and / or Component (B) may be about 0.01 to 100 mg / kg, preferably about 0.1 to 10 mg / kg, more preferably about 0.5 to 5 mg / kg.

In particular, the dose (intake, dose) of component (A) is 0.01 to 100 mg / kg, preferably 0.1 to 10 mg / kg, more preferably 0.5 to 5 mg / kg per day. It may be a degree.

In the case of administering the osteoclastic extract, the dosage (intake, dose) of the osteoclastic extract is 10 to 2000 mg / kg, preferably 20 to 1000 mg / kg, more preferably 50 to 100 per day. It may be about 800 mg / kg (for example, 100 to 700 mg / kg).

In addition, the dose (intake, dose) of component (B) (such as Ezokogi extract) is 10 to 2000 mg / kg, preferably 20 to 1000 mg / kg, more preferably 50 to 800 mg / kg per day ( For example, it may be about 100 to 700 mg / kg).

Further, when the components (C) are administered in combination, the dose (intake, dose) of the component (C) is 0.01 to 100 mg / kg, preferably 0.1 to 10 mg / kg per day. More preferably, it may be about 0.5 to 5 mg / kg.

Administration may be divided once or multiple times.

The agent or composition of the present invention can be provided in the form of a kit if desired. Such a kit may be, for example, a kit comprising a container such as a pack or a dispenser device that may contain one or more unit dosage forms containing component (A) and / or component (B). Good.

In the present invention, separate agents or compositions can be combined in a kit form. For example, such a kit may be composed of a container containing an agent or composition containing component (A) and a container containing an agent or composition containing component (B).

Such kits typically include containers for containing separate agents or compositions, such as split bottles or split foil packets, but include the separate compositions in a single unsplit container. You can also. Kit forms are particularly useful when different components are administered in different dosage forms, when separate components are administered at different dosage intervals, or when the combined individual components need to be titrated by the prescribing physician. is there.

Packs can include, for example, metal or plastic foil, blister packs and the like. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packs generally consist of a sheet of relatively hard material covered by a foil of transparent plastic material. During the packaging process, recesses are formed in the plastic foil. These indentations are tailored to the size and shape of the individual tablets or capsules to be packed. The tablet or capsule is then placed in the recess and the sheet of relatively hard material is sealed against the plastic foil at the foil surface opposite to the direction in which the recess was formed. As a result, the tablets or capsules are sealed in the recesses between the plastic foil and the sheet. The strength of the sheet is preferably such that the tablet or capsule can be removed from the blister pack by manually applying pressure to the recess so that an opening is formed in the sheet at the location of the recess. Tablets or capsules can be removed through the opening.

* Package or dispenser device can be accompanied by package inserts, product inserts, etc. for administration. Containers such as packs or dispensers can be adapted to the notifications of government agencies and authorities that regulate the manufacture, use or sale of medicines.

<Food &Drink>
The present invention includes foods and drinks containing the component (A), the component (B) (and other components such as the component (C)) (that is, foods and drinks containing the agent or composition).
In such a food and drink, preferred embodiments of the component (A) and the component (B), the ratio thereof, and the like are the same as described above.

In addition to general foods, including so-called health foods, foods and drinks also include health foods such as foods for specified health use and foods with nutrient function specified by the Ministry of Health, Labor and Welfare. Nutritional supplements), feed, food additives, and the like are also included in the food and drink of the present invention.

In addition, the food and drink may be food or drink for a specific subject [for example, for elderly people, patients or patients (for example, for patients having a neurological disease (such as the above-described neurological diseases)]). Good.

In order to use the component (A) and / or the component (B) in foods and drinks, it is necessary to produce foods and drinks as they are or together with various nutritional components and the like in raw materials for foods and drinks such as processed meat and soft drinks. it can.

In addition, when the components (A) and / or (B) are used as health foods, dietary supplements, etc., for example, using conventional means, tablets, capsules (soft capsules, hard capsules, etc.), powders, granules, liquids (Suspensions, syrups, etc.), emulsions, jellies, sticks, etc. The tablet includes a disintegrating tablet (orally disintegrating tablet).

The food and drink may contain a food additive (food additive) as long as it contains bone fracture supplement or an extract of bone fracture supplement. Although it does not specifically limit as a food additive, For example, excipient | fillers (for example, wheat starch, corn starch, cellulose, lactose, sucrose, mannitol, sorbitol, xylitol, pregelatinized starch, casein, magnesium aluminate silicate, Calcium silicate, etc.), binder (eg, pregelatinized starch, hydroxypropylmethylcellulose, polyvinylpyrrolidone, etc.), disintegrant (eg, cellulose, hydroxypropylcellulose, corn starch, etc.), fluidizing agent (eg, light anhydrous silicic acid) Sucrose fatty acid esters, etc.), oils (eg, vegetable oils such as soybean oil, sesame oil, olive oil, linseed oil, sesame oil, rapeseed oil, coconut oil, corn oil, etc.), nutrients (eg, various Minera , Various vitamins, amino acids), flavorings, sweeteners, flavoring agents, coloring agents, solvents (ethanol), salts, surfactants, pH regulators, buffers, antioxidants, stabilizers, gelling agents, thickening Agents, lubricants, encapsulating agents, suspending agents, coating agents, preservatives and the like.

Food additives may be used alone or in combination of two or more, and in particular, two or more may be used. When two or more kinds of additives are used, two or more components (for example, two or more components selected from the components classified as excipients) may be selected from the same type of additives. May be combined (for example, a combination of an excipient and a binder), or both of them may be selected. In particular, the additive may be a combination of different additives.

In addition, in the food and drink in the form of tablets and the like, at least two or more additives selected from excipients, binders, and disintegrants may be used.

When the component (A) and / or the component (B) is used as a food additive, the food or drink is not particularly well-maintained. However, for example, food [e.g., noodles (such as soba, udon, Chinese noodles, instant noodles), Confectionery (cake, candy, gum, chocolate, snacks (potato chips, etc.), biscuits, cookies, gummy, jelly, jam, butter, cream (cream puffs, cakes, etc.), breads, marine or livestock processed foods (kamaboko) , Ham, sausage, etc.), dairy products (processed milk, fermented milk, etc.), fats and oils and processed foods (salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing, etc.), seasonings (sauce, sauce, etc.) , Retort food (curry, stew, rice cake, porridge, miscellaneous food, etc.), frozen dessert (ice cream, sha Bet, etc. shaved ice), fried (croquettes, fries, etc. fried chicken), etc.], drinks (tea drinks, soft drinks, carbonated drinks, energy drinks, fruit drinks, such as lactic acid beverage), and the like.

The amount of component (A) and / or component (B) in the above-mentioned food and drink varies depending on the addition form and administration form, and can be selected from a wide range, for example, 0.001 to 90% by weight, preferably 0. 001 to 80% by weight, more preferably 0.1 to 70% by weight (eg 1 to 50% by weight), usually 0.01 to 50% by weight (eg 0.1 to 30% by weight) Also good.

In particular, when the food or drink of the present invention contains a bone crush supplement, the proportion of the bone crush extract is 1 to 90% by weight, preferably 10 to 70% by weight, more preferably about 20 to 50% by weight. May be.

In addition, particularly when the food or drink of the present invention contains an elephant extract, the ratio of the extract is, for example, 1 to 90% by weight, preferably 10 to 70% by weight, more preferably about 20 to 50% by weight. May be.

Furthermore, when the food or drink of the present invention contains the component (C), the proportion of the component (C) is, for example, 0.001 to 90% by weight, preferably 0.001 to 80% by weight, more preferably 0.1 to 70% by weight (for example, 1 to 50% by weight), usually about 0.01 to 50% by weight (for example, 0.1 to 30% by weight), or about 1 to 90% by weight It may be.

In addition, in food and drink, the dose (or intake or dose) of component (A) and / or component (B) (and component (C)) can be selected from the same range as described above.

EXAMPLES The present invention will be described more specifically with reference to the following examples. However, the present invention is not limited to these examples, and many modifications are within the technical scope of the present invention. This is possible by those with ordinary knowledge.

In addition, in the Example, the bone crushing extract (bone crushing water extraction) used what was obtained as follows.

Twenty times the amount of water was added to osteoclast (Tochimoto Tenkaido, Osaka), and extraction was performed using a decoction device (Uchida Wakayaku, Tokyo). The mixture was heated on a strong fire until the water added to the crude drug boiled (15-20 minutes), and then the heat was continued for 1 hour. The filtrate was filtered and then filtered with a cotton plug while hot. The filtrate was allowed to cool at room temperature and then frozen with liquid nitrogen. Furthermore, it was made to dry with a freeze dryer, and the extract of the powdered osteoclastic supplement was obtained (yield 7.8%).

In the examples, the object recognition memory test was performed as follows.
<Object recognition memory test>
The object recognition memory test is a test using a habit of showing interest in new things by animals. That is, in the test stage, it is a test for confirming whether or not the object seen in the training stage is remembered. The test was performed as described in the literature (Int J Neurosci, 121, pp.181-190, 2011. and Int J Neurosci, 121, pp.641-648, 2011.). Specifically, it was performed as follows.

The test was conducted in a relatively well-lit room (approximately 100 lux). Appropriate time intervals between the training phase and the test phase were determined in advance by testing with different groups of mice. There are no landmarks on the inner wall of the open field box under test. In the training phase, place two identical objects in the field and let them search for 10 minutes. During the test phase, one of the objects is replaced with a new object, but the place is not changed, and a 10-minute exploratory action is performed. The increase in the number of times the mouse performs an exploratory action with interest in the replaced new object is used as an index of the object memory ability.
In the example, the ratio (%) of the number of searches for a new object with respect to the total search time is calculated as a search index (Preference index).

Example 1
Naringenin and naringenin-7'-O-glucuronide, one of the glycosides of naringenin, were synthesized in order to investigate the activity against nerve cells. First, projections on normal cortical neurons The extension action was examined. On the third day of culture of primary cultured mouse cerebral cortical neurons, each compound was treated at a concentration of 0.01, 0.1, 1, 10 μM, and after 4 days, the cells were fixed, and then axons (pNF− The length per cell (H positive) was examined by immunostaining.

The structures of these two compounds and naringenin-4′-O-glucuronide described later are shown below.

The results are shown in FIG. In FIG. 1, “Cont” indicates control, “Naringenin” indicates naringenin, and “Naringenin-7-O” indicates naringenin-7′-O-glucuronide. The value is 100 μm / neuron. It is converted.

As is clear from the results of FIG. 1, it was found that both naringenin and naringenin-7'-O-glucuronide showed activity.

(Example 2)
In Example 1, in addition to the synthesized naringenin and naringenin-7′-O-glucuronide, naringenin-4′-O-glucuronide, which is a glycoside of naringenin, was synthesized, and axon of Aβ (25-35) was synthesized. The stretching action was examined as follows.

On the third day of culture of primary cultured mouse cerebral cortical neurons, 10 μM Aβ (25-35) was treated, and three days later, each compound was treated at concentrations of 0.01, 0.1, 1, and 10 μM. After 4 days, the cells were fixed, and the length per cell of the axon (pNF-H positive) was examined by immunostaining.

The results are shown in FIG. In FIG. 2, “Cont” is the control (no treatment of Aβ (25-35)), “Veh” is the solvent only, “Naringenin” is naringenin, “Naringenin-4′-O” is naringenin-4′- O-glucuronide, “Naringenin-7-O” indicates naringenin-7′-O-glucuronide, and the value is converted to μm / neuron with the control value being 100.

As is apparent from the results of FIG. 2, re-extension activity of any of naringenin, naringenin-4′-O-glucuronide and naringenin-7′-O-glucuronide against axonal atrophy caused by Aβ (25-35) Was recognized.

In the following examples, osteoprosthesis known to contain naringenin was examined.

(Example 3)
After the novel object recognition test, each mouse was anesthetized and transcardially perfused with chilled saline. The whole brain was carefully removed from the skull, soaked in 4% paraformaldehyde overnight, then soaked in 30% sucrose-PBS and stored at −30 ° C. Brains were cut into 20 μm continuous coronal sections using a cryostat (CM3050S, Leica, Heidelberg, Germany).

Slices were fixed with 4% paraformaldehyde and polyclonal antibodies against Aβ (1-40 / 42) (1: 300) (Chemicon, Temecula, Calif., USA), pNF-H (1: 300). ) And a monoclonal antibody (Covance, Emeryville, CA, USA) for 20 hours at 4 ° C. As secondary antibodies, Alexa Fluor 488-labeled goat anti-rabbit IgG antibody (1: 300) and Alexa Fluor 568-labeled goat anti-mouse antibody (1: 300) (Molecular Probes, Eugene, Oregon) , USA).

Fluorescence images of axons and Aβ (1-40 / 42) were taken at 324 μm × 430 μm using a fluorescence microscope (BX-61).

The area of extracellular amyloid plaques was measured using image analysis software ImageJ (http://rsbweb.nih.gov/ij). The area of pNF-H positive hypertrophied and axon terminal was measured using ImageJ for all amyloid plaques.

The bone replenishment extract was dissolved in physiological saline and orally administered once daily to 5XFAD mice (6-8 months old, male and female) at a dose of 500 mg / kg / day for 31 days. The amount of the drug solution to be administered orally was 10 ml / kg body weight.

The results are shown in Figs. In FIGS. 3 to 10, “Wild” is a wild-type mouse, “5XFAD” is “5XFAD mouse”, “Veh” is a solvent only, “Ext500” is a group administered with a bone replenishment extract at a rate of 500 mg / kg (Mouse).

As is apparent from the results of FIGS. 3 to 10, amyloid plaques are not observed in wild-type mice in any part of the prefrontal cortex, periorbital cortex, hippocampal CA1, and hippocampal dentate gyrus, whereas 5 × FAD mice It was increasing.

On the other hand, the amyloid plaque area significantly decreased in the osteoclastic rehydration extract 500 mg / kg administration group. Moreover, when the area of the enlarged terminal end of the enlarged axon observed in the vicinity of the amyloid plaque was quantified, the degenerated axon was wild in any part of the prefrontal cortex, periolfactory cortex, hippocampal CA1, and hippocampal dentate gyrus. It was not observed in type mice, but increased in 5XFAD mice. In the osteoclastic rehydration extract 500 mg / kg administration group, a tendency to decrease this was shown.

Example 4
The bone replenishment extract was dissolved in physiological saline and orally administered to 5XFAD mice (45 weeks old, male) once a day at a dose of 500 mg / kg / day. A 5XFAD mouse group (45 weeks old, male) to which only the solvent was administered and a wild type mouse group (45 weeks old, male) to which only the solvent was administered were also provided.

The amount of drug solution to be administered orally was 10 ml / kg body weight. The administration period was 22 days. An object recognition memory test was performed on these mice, and after confirming that a memory improving effect was observed in the bone-fed rehydration extract administered group, the cerebral cortex of the mice was extracted after refluxing and lysate was prepared.

The direct binding protein search experiment of the osteoclast rehydration extract found the involvement of Collapsin response mediator protein-2 (CRMP2, also known as dihydropyrimidinase-related protein 2, TOAD-64) protein prior to this example.

The degree of CRMP2 phosphorylation in cerebral cortical lysate was examined by Western blotting. The antibody used was an antibody that detects phosphorylation of threonine of the 514th amino acid of CRMP2.

The results are shown in FIG. In FIG. 11, “Wild” is a wild-type mouse, “5XFAD” is “5XFAD mouse”, “Veh” is a solvent only, and “Ext” is a group administered with a crushed rehydration extract at a rate of 500 mg / kg ).

As is clear from FIG. 11, CRMP2 phosphorylation was enhanced in the 5XFAD mouse solvent-administered group compared to wild-type mice, whereas CRMP2 phosphorylation was significantly decreased in the 5XFAD mouse administered with the crushed water extract. did.
Reduction of CRMP2 phosphorylation has already been recognized as a mechanism that leads to promotion of axonal extension. Therefore, suppression of CRMP2 phosphorylation in the mouse brain by administration of a bone replenishing water extract is associated with axonal elongation and It is clear that this is part of the mechanism of memory enhancement.

(Example 5)
The components contained in the osteoclastic prosthesis were analyzed, and which components can contribute to axonal extension was examined as follows.

First, the bone replenishing water extract was dissolved in sterilized purified water. Mouse cerebral cortical neurons were prepared from embryonic day 14 ddY mice.

The crushed rehydration extract was further fractionated into a petroleum ether fraction, an ethyl acetate fraction, an n-butanol fraction, and a water-soluble fraction.

On day 3 of culture of primary cultured mouse cerebral cortical neurons, 10 μMAβ (25-35) was treated, 3 days later, each fraction was treated at a concentration of 1 or 10 μg / mL, and another 4 days later After fixing the cells, the length per cell of axons (pNF-H positive) and dendrites (MAP2 positive) was examined by immunostaining.

The results are shown in FIG. In FIG. 12, “Cont” is the control, “Veh” is the solvent only, “Pe” is the petroleum fraction, “EtOAc” is the ethyl acetate fraction, “BuOH” is the n-butanol fraction, “Water”. Indicates a water-soluble fraction, and the value is converted to μm / neuron with a control value of 100.

As is apparent from FIG. 12, the axon density was significantly decreased in Aβ25-35 cells compared to Aβ25-35 untreated cells. Axon extension tendency was observed in all fractions, but axons were significantly extended especially by the post-treatment of the n-butanol fraction (10 μg / mL).

(Example 6)
The n-butanol fraction obtained in Example 4 was analyzed in detail, and the components that can contribute to axonal extension at the compound level were examined as follows.

First, (A) n-butanol fraction was subdivided with silica gel to isolate five compounds, which were naringenin (1), neoeriocitrin (2), 5,7-dihydroxychromone-7-O-neohe It was identified from the results of LC-MS mass analysis that it was spidoside (4) and protocatechuic acid (5).

The LC-MS chart of the n-butanol fraction is shown in FIG. 13, and the structure of each identified compound is shown in the following formula.

Then, 10 μM Aβ (25-35) was treated on the 3rd day of culture of primary cultured mouse cerebral cortical neurons, and 3 days later, 5 compounds each were treated at a concentration of 1 or 10 μM. After fixing the cells one day later, the length per cell of axons (pNF-H positive) and dendrites (MAP2 positive) was examined by immunostaining.

The results are shown in FIG. In FIG. 14, “Cont” is the control, “Veh” is the solvent only, numbers such as “1” indicate each compound, and the value is the value of the control, which is converted to μm / neuron.

As is apparent from FIG. 14, the axon density was significantly decreased in the Aβ25-35 cells compared to the Aβ25-35 untreated cells, but the treatment with the compounds 1 to 5 extended the axons. . In particular, neoeriocitrin (compound 2), which is a glycoside of eriodictyol, and compound 4, which is a glycoside of caffeic acid, showed a remarkable axon extension effect.

(Example 7)
Dissolve bone replenishment extract in physiological saline and administer once a day at a dose of 500 mg / kg / day, or dissolve elephant leaf water extract in physiological saline once a day as 500 mg / kg / day, Alternatively, a bone replenishing water extract (500 mg / kg / day) and an Ekogi leaf water extract (50 mg / kg / day) were orally administered to 5XFAD mice once a day at the same time.

In addition, the Ezoukogi leaf extract was used by freeze-drying a filter obtained by infiltrating the leaves of Eleutherococcus senticosus in the family Urugiaceae into water at 85 degrees for 30 minutes and filtering.

5XFAD mice were female, 27-56 weeks old, and wild-type mice in the control group were female, 52 weeks old. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 15 days. An object recognition memory test was performed on these mice. The interval between the training stage and the test stage was set to two conditions of 1 hour (Test 1) and 24 hours (Test 2).

The results are shown in FIG. In FIG. 15, “Wild” is a wild-type mouse, “5XFAD” is “5XFAD mouse”, “Veh” is a solvent alone, “Kotsusaiho” is a group to which only a bone rehydration extract is administered, “Ezoukogi” is an Ekoukogi leaf A group to which only the water extract was administered, “Kotsu / Ezoukogi” represents a group to which both the bone replenishing water extract and the Ezoukogi leaf water extract were administered.

As is clear from the results of FIG. 15, object recognition memory was maintained in both test 1 and test 2 in the wild type mouse, whereas memory impairment occurred in the solvent administration group of the 5XFAD mouse.

On the other hand, in the osteoclastic rehydration extract alone administration group and the Ezoukogi leaf water extract administration group, memory impairment was improved when the interval time was as short as 1 hour, but test 2 with an interval time of 24 hours showed an improvement in memory. I can't. On the other hand, in the simultaneous administration group of the crushed water replenishing extract and the Ezoukogi leaf water extract, both the test 1 and the test 2 showed significant improvement in memory impairment.

This result shows that the osteoclast replenishment extract and Ezoukogi leaf water extract alone have a memory improvement effect, but the combined use of the osteoclast rehydration extract and Ezoukogi leaf water extract has a synergistic and stronger memory improvement effect. It shows that it is obtained.

(Example 8)
Ezoukogi leaf water extract was dissolved in physiological saline and administered orally to normal mice (ddY, male, 7 weeks old) once a day at a dose of 500 mg / kg / day. The amount of the drug solution to be administered orally was 10 mL / kg body weight. The administration period was 27 days.
In addition, Ezoukogi leaf water extract was used by freeze-drying a leaf of Eleutherococcus senticosus belonging to the family Urugiaceae infiltrated with water at 85 degrees for 30 minutes.
An object recognition memory test was performed on these mice. Training was performed on the 24th day after administration, and the test was performed on the 27th day after administration. The training and test interval was 72 hours.
The results are shown in FIG. In FIG. 16, “Control” and a control group to which only the solvent is administered, “Extract” is an Ezocogi leaf water extract administration group, “Training” is training, and “Test” is a test.
As is clear from the results in FIG. 16, the object recognition memory of normal mice was significantly enhanced in the Ezocogi leaf water extract administration group compared to the control group to which only the solvent was administered.

In the present invention, an agent or composition useful for improving or improving memory ability and cognitive function can be obtained.

Claims (24)

1. Including at least one component (A) or sorghum component (B) selected from naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof, and salts thereof, An agent for use in at least one application selected from the following (1), (2) and (3).
   (1) Restoration and / or extension of neurites (2) Improvement or improvement of memory and / or cognitive function (3) Prevention and / or treatment of neurological diseases (however, it contains only naringenin as an active ingredient and prevents Alzheimer's disease) And / or excluding therapeutic agents)
2. The agent according to claim 1, wherein the component (A) comprises at least one component selected from naringenin, a glycoside of naringenin, and salts thereof.
3. The agent according to claim 1 or 2, wherein the component (A) comprises at least one component selected from naringenin, naringin, naringen glucuronide, and pharmaceutically acceptable salts thereof.
4. The agent according to any one of claims 1 to 3, wherein the component containing the component (A) comprises a bone crushing auxiliary component.
5. The agent according to any one of claims 1 to 4, wherein the component (A) and the component (B) are used in combination.
6. The agent according to any one of claims 1 to 5, comprising a component (A) and a component (B).
7. The component (A) contains a bone crush extract, and the component (B) contains a carp extract, and the ratio of the bone crush extract and the carp extract is the former / the latter (weight ratio) = 1 / 0.01-1 The agent according to claim 5 or 6, which is / 100.
8. Furthermore, the agent according to any one of claims 1 to 7, which is used in combination with at least one component (C) selected from diosgenins, dihydroxyvitamin D3 and desonomin.
9. The agent according to claim 8, wherein the ratio of component (A) and / or component (B) to component (C) is the former / the latter (weight ratio) = 1 / 0.01 to 1/100.
10. At least one component (A) selected from naringenin, eriodictyol, 5,7-dihydroxychromone, caffeic acid, protocatechuic acid, glycosides thereof, and salts thereof, and the sorghum component (B) A composition comprising.
11. The composition according to claim 10, wherein the component (A) comprises at least one component selected from naringenin, a glycoside of naringenin, and salts thereof.
12. The composition according to claim 10 or 11, wherein the component (A) comprises at least one component selected from naringenin, naringin, naringen glucuronide, and salts thereof.
13. The composition according to any one of claims 10 to 12, comprising an osteoclastic component as a component containing the component (A).
14. The component (A) contains a bone crush extract, and the component (B) contains a carp extract, and the ratio of the bone crush extract and the carp extract is the former / the latter (weight ratio) = 1 / 0.01-1 The composition according to any one of claims 10 to 13, which is / 100.
15. Furthermore, the composition according to any one of claims 10 to 14, which is used in combination with at least one component (C) selected from diosgenins, dihydroxyvitamin D3 and desonomin.
16. 16. The composition according to claim 15, wherein the ratio of component (A) and / or component (B) to component (C) is the former / the latter (weight ratio) = 1 / 0.01 to 1/100.
17. The composition according to any one of claims 10 to 16, which is a composition for use in at least one application selected from the following (1), (2) and (3).
    (1) Restoration and / or extension of neurites (2) Improvement or improvement of memory and / or cognitive function (3) Prevention and / or treatment of neurological diseases
18. A food or drink containing the agent or composition according to any one of claims 5 to 16.
19. The food or drink according to claim 18, which is a supplement, health food, functional labeling food, nutritional functional food or food for specified health use.
20. The food or drink according to claim 18 or 19, comprising 1 to 90% by weight of a bone fracture supplement.
21. The food or drink according to any one of claims 18 to 20, comprising 1 to 90% by weight of Ezoukogi extract.
22. It contains 1-90% by weight of the osteoclastic extract and 1-90% by weight of the elephant extract, and the ratio of the osteoclastic extract to the elephant extract is the former / the latter (weight ratio) = 1 / 0.01 to 1/100. The food or drink according to any one of claims 18 to 21.
23. The food or drink according to any one of claims 18 to 22, further comprising at least one component (C) selected from diosgenins, dihydroxyvitamin D3, and desonomin.
24. The food / beverage product according to claim 23, wherein the ratio of component (A) and / or component (B) to component (C) is the former / the latter (weight ratio) = 1 / 0.01 to 1/100.

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