Pharmaceutical composition for preventing, ameliorating or treating diseases associated with brain cognitive function, food composition for improving brain cognitive function, feed composition for improving brain cognitive function, and method for preparing same

Abstract

The present invention relates to a pharmaceutical composition for preventing, ameliorating or treating diseases associated with brain cognitive function, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for preparing same and, specifically, to: a pharmaceutical composition for preventing, ameliorating or treating diseases associated with brain cognitive function, a food composition for improving brain cognitive function, and a feed composition for improving brain cognitive function, each comprising, as active ingredients, extracts of Atractylodes lancea, Eclipta prostrata, and Magnolia officinalis, and a method for preparing same; and a pharmaceutical composition for preventing, ameliorating or treating diseases associated with brain cognitive function, a food composition for improving brain cognitive function, and a feed composition for improving brain cognitive function, each comprising, as active ingredient(s), atractylodin, wedelolactone, and / or honokiol, and a method for preparing same.

Description

Pharmaceutical composition for preventing, improving, or treating diseases related to brain cognitive function, food composition for improving brain cognitive function, feed composition for improving brain cognitive function, and method for manufacturing the same

[0001] The present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same. Specifically, the invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark as active ingredients; and a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising Atractylodin, Wedelolactone, and / or Honokiol as active ingredients.

[0002] In modern society, aging, stress, and changes in lifestyle are rapidly increasing the incidence of various brain cognitive function-related diseases, such as Alzheimer's disease, mild cognitive impairment (MCI), and dementia. These diseases not only reduce the quality of life for patients but also cause serious economic and emotional burdens on families and society as a whole.

[0003] Major pathological causes associated with the decline in brain cognitive function include neurotransmitter imbalance, oxidative stress, inflammatory responses, protein aggregation, and brain cell damage. For example, a decrease in acetylcholine has been identified as a major issue in Alzheimer's disease, and acetylcholinesterase inhibitors have been developed to regulate this. Additionally, drugs targeting NMDA receptors are used to prevent neuronal damage caused by glutamate excess, and drugs that regulate dopamine levels or stimulate receptors are utilized in the treatment of Parkinson's disease.

[0004] Oxidative stress and inflammatory responses are another major cause of cognitive decline. The accumulation of reactive oxygen species (ROS) damages neurons, which accelerates the progression of neurodegenerative diseases. Antioxidants are being studied to prevent this damage, and drugs are being developed to protect brain cells by regulating inflammatory cytokines or inhibiting the overactivation of microglia.

[0005] Protein aggregation is also closely related to the decline in cognitive function. Beta-amyloid plaques and tau protein aggregations in Alzheimer's disease, as well as alpha-synuclein aggregates in Parkinson's disease, interfere with neuronal function, and active research is underway on protein aggregation inhibitors or antibody therapies targeting these factors.

[0006] While currently used cognitive function-enhancing drugs primarily focus on symptom relief, new treatments are being developed to fundamentally prevent neuronal damage, enhance neural circuit plasticity, and promote neuronal regeneration. Furthermore, gene therapy and neural stem cell-based therapies are garnering attention, and these treatments are maximizing their effectiveness when combined with non-pharmacological approaches such as cognitive training, dietary adjustments, and exercise.

[0007] Under these circumstances, pharmaceutical and food compositions based on naturally derived ingredients are attracting attention as alternatives for the prevention and treatment of brain cognitive function-related diseases, based on their high safety and diverse physiological activities.

[0008] The present invention provides pharmaceutical and food compositions effective for preventing and improving the decline of brain cognitive function by including such natural extracts as active ingredients, and further proposes a new paradigm for the treatment and prevention of brain cognitive function-related diseases by presenting a manufacturing method that combines safety and efficacy.

[0009] Furthermore, with the recent rapid increase in the pet-owning population, interest in the health and welfare of pets is rising, and the status of pets is shifting to that of family members.

[0010] In particular, as the average lifespan of companion animals increases, various age-related diseases in elderly companion animals are emerging as important subjects for research and management. Among these diseases, cognitive decline and dementia are identified as major causes that significantly reduce the quality of life of companion animals. These diseases not only reduce the quality of life of companion animals but also cause emotional burdens for owners and the household.

[0011] It is known that Cognitive Dysfunction Syndrome (CDS), characterized by symptoms such as cognitive decline, memory loss, disorientation, and behavioral changes, occurs frequently in companion animals, particularly dogs and cats, as they age. This condition is reported to have pathological mechanisms similar to those of Alzheimer's disease in humans, involving a complex interplay of factors including neuronal damage, beta-amyloid deposition, increased oxidative stress, and neuroinflammatory responses.

[0012] Major pathological causes associated with cognitive decline include neurotransmitter imbalances, oxidative stress, inflammatory responses, protein aggregation, and brain cell damage. For example, cognitive decline in aging dogs has been identified as a primary cause of neurotransmitter imbalances, such as a decrease in acetylcholine; while drugs and supplements are being developed to regulate this, effective treatments remain limited.

[0013] In addition, oxidative stress and inflammatory responses are other major causes that can exacerbate cognitive decline in companion animals. The accumulation of reactive oxygen species (ROS) damages brain cells, which can accelerate cognitive impairment. Antioxidants are being studied to prevent this damage, and drugs are being developed to protect brain cells by regulating inflammatory cytokines or inhibiting the overactivation of microglia.

[0014] Protein aggregation is also closely related to the decline in cognitive function in companion animals. Cognitive impairment in older dogs exhibiting pathological features similar to Alzheimer's disease may be caused by beta-amyloid plaques or tau protein aggregation interfering with brain cell function. Consequently, active research is underway on protein aggregation inhibitors and antibody therapies targeting these factors.

[0015] Currently used cognitive function-enhancing drugs for companion animals primarily focus on symptom relief. While new treatments are being developed to fundamentally prevent neuronal damage, enhance neural circuit plasticity, and promote neuronal regeneration, they remain limited in their ability to fundamentally inhibit disease progression or achieve recovery. Particularly for companion animals, a differentiated approach compared to human treatments is required due to concerns regarding drug safety and side effects from long-term administration.

[0016] Under these circumstances, pharmaceutical and feed compositions based on naturally derived ingredients are attracting attention as alternatives for the prevention and treatment of brain cognitive function-related diseases in companion animals, based on their high safety and diverse physiological activities.

[0017] Furthermore, as pet owners' awareness of brain cognitive function-related diseases in companion animals expands, the demand for early diagnosis and personalized treatment is increasing. Consequently, the need for developing new drugs or feed ingredients that can effectively improve or protect the brain cognitive function of companion animals is being further emphasized.

[0018] The present invention provides pharmaceutical and feed compositions effective for preventing and improving the decline of brain cognitive function in companion animals by including such natural extracts as active ingredients, and further proposes a new paradigm for the treatment and prevention of cognitive dysfunction syndrome in companion animals by presenting a manufacturing method that combines safety and efficacy.

[0019] The inventors of the present invention have made diligent research efforts to develop a composition for improving brain cognitive function. As a result, the present invention was completed by identifying that extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis are effective in improving brain cognitive function.

[0020] In addition, the present invention was completed by identifying that Atractylodin, Wedelolactone, and Honokiol are effective in improving brain cognitive function.

[0021] Accordingly, the objective of the present invention is to provide a pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0022] Another objective of the present invention is to provide a composition for improving brain cognitive function comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0023] Another objective of the present invention is to provide a method for preparing a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0024] Another objective of the present invention is to provide a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for the prevention, improvement, or treatment of brain cognitive function-related diseases.

[0025] Another objective of the present invention is to provide a use for improving brain cognitive function of a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0026] Another objective of the present invention is to provide a method for preventing, improving, or treating brain cognitive function-related diseases using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0027] Another objective of the present invention is to provide a method for improving brain cognitive function using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0028] Another objective of the present invention is to provide a pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, comprising attractilodine, wedelolactone, and / or honokiol as active ingredients.

[0029] Another objective of the present invention is to provide a composition for improving brain cognitive function comprising attractylodin, wedelolactone and / or honokiol as active ingredients.

[0030] Another object of the present invention is to provide a method for preparing a composition comprising attractylodin, wedelolactone and / or honokiol as an active ingredient.

[0031] Another objective of the present invention is to provide a composition comprising attractilodine, wedelolactone and / or honokiol as an active ingredient for the prevention, improvement, or treatment of brain cognitive function-related diseases.

[0032] Another objective of the present invention is to provide a use for improving brain cognitive function of a composition comprising attractylodin, wedelolactone and / or honokiol as an active ingredient.

[0033] Another objective of the present invention is to provide a method for preventing, improving, or treating brain cognitive function-related diseases using a composition comprising atractillodin, wedelolactone and / or honokiol as active ingredients.

[0034] Another objective of the present invention is to provide a method for improving brain cognitive function using a composition comprising attractylodin, wedelolactone and / or honokiol as active ingredients.

[0035] The inventors have made diligent research efforts to develop a composition for improving brain cognitive function in companion animals. As a result, the present invention was completed by identifying that extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis are effective in improving brain cognitive function in companion animals.

[0036] Accordingly, the objective of the present invention is to provide a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases in companion animals, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0037] Another objective of the present invention is to provide a feed composition for improving brain cognitive function in companion animals, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0038] Another objective of the present invention is to provide a method for preparing a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0039] Another objective of the present invention is to provide a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for the prevention, improvement, or treatment of brain cognitive function-related diseases in companion animals.

[0040] Another objective of the present invention is to provide a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for improving brain cognitive function in companion animals.

[0041] Another objective of the present invention is to provide a method for preventing, improving, or treating brain cognitive function-related diseases in companion animals using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0042] Another objective of the present invention is to provide a method for improving brain cognitive function in companion animals using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0043] The present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same. Specifically, the invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark as active ingredients; and a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising Atractylodin, Wedelolactone, and / or Honokiol as active ingredients.

[0044] The present invention will be described in more detail below.

[0045] One example of the present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0046] In this specification, the term "containing as an active ingredient" means containing an amount sufficient to achieve the efficacy or activity of the extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark.

[0047] The term "pharmaceutical effective amount" in this specification means an amount sufficient to achieve the efficacy or activity of the aforementioned extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark.

[0048] In this specification, the term "improvement" refers to any act in which symptoms of brain cognitive function-related diseases are improved or completely cured by the administration of the composition according to the present invention.

[0049] In this specification, the term "prevention" refers to any act of suppressing or delaying the symptoms of brain cognitive function-related diseases through the administration of a composition according to the present invention.

[0050] In this specification, the term "treatment" refers to any act in which symptoms of brain cognitive function-related diseases are improved or completely cured by the administration of a composition according to the present invention.

[0051] The term "Changchul (Atractylodes lancea and Atractylodes chinensis)" in this specification refers to a perennial herbaceous plant belonging to the Asteraceae family, which has long been used for medicinal purposes, mainly in East Asia. The rhizome of this plant is used in a dried form for medicinal purposes, and its efficacy is widely recognized in Eastern medicine.

[0052] Changchul grows to a stem height of approximately 30 to 100 cm. Its leaves are simple or deeply lobed, and its flowers feature a small capitulum structure characteristic of the Asteraceae family. It primarily grows wild in mountainous regions and prefers well-drained soil and semi-shaded environments. It is also resistant to cold and is relatively easy to cultivate and manage. To be used as a medicinal herb, the rhizomes are harvested in the autumn, washed, and dried.

[0053] The main medicinal effects of Atractylodes are strengthening the digestive organs (spleen and stomach), promoting digestive function, and eliminating excess moisture accumulated in the body. In Korean traditional medicine, it is widely used to improve digestive disorders such as loss of appetite, diarrhea, and abdominal bloating. It is also known to be effective in relieving body heaviness, fatigue, and joint pain caused by a damp environment.

[0054] Interest in Atractylodes lanceolata is also growing in modern medical research. Due to its anti-inflammatory, immune-modulating, and antioxidant effects, Atractylodes lanceolata is being suggested as a potential candidate for the prevention and treatment of chronic diseases. These efficacyes are being confirmed in various studies aiming to scientifically validate its traditional medicinal value.

[0055] In the present invention, the atractylodes may be one or more selected from the group consisting of *Atractylodes lancea DC* and *Atractylodes chinensis Koidzumi*, but is not limited thereto.

[0056] In the present invention, the plant may be one or more selected from the group consisting of leaves, stems, fruits, roots, and flowers, and may be, for example, roots, but is not limited thereto.

[0057] The term "Eclipta prostrata" in this specification refers to an annual or perennial herb belonging to the Asteraceae family. It grows naturally in tropical and subtropical regions, including East Asia, and is widely used for medicinal and edible purposes. In Korean, it is also called Mukhanryeon or Yeonjacho, and is regarded as an important medicinal herb in traditional medicine. Eclipta prostrata grows to a height of about 20 to 60 cm and has stiff hairs on its leaves and stems. The leaves are opposite and willow-leaf shaped, measuring about 3 to 9 cm in length and 5 to 15 mm in width. The stems grow upright or lying obliquely and have the tendency to branch out from every leaf axil. Small, white, capitulum-shaped flowers bloom from summer to autumn; after the flowers wither, oval or round fruits form, containing small brown seeds inside.

[0058] This plant thrives in fertile, well-drained soil and grows rapidly. It is primarily harvested in the summer and autumn, and the entire plant can be used for medicinal purposes. In traditional medicine, *Hallyeocho* is known to protect the liver and kidneys, improve blood circulation, and support hair and skin health. In particular, it is effective in alleviating symptoms of graying or hair loss, and is also used to treat eye diseases or jaundice caused by liver dysfunction. Furthermore, due to its antipyretic, hemostatic, and detoxifying properties, it is useful for wound healing and reducing inflammation.

[0059] According to current research, *Hallyeocho* contains bioactive substances such as flavonoids, coumarins, and wedelolactone, and these components have been found to exhibit anti-inflammatory, anticancer, antibacterial, and antioxidant effects. In particular, it is highly effective in reducing inflammation and oxidative stress, and some studies have also confirmed its ability to inhibit cancer cell proliferation.

[0060] As such, the water hyacinth is a versatile plant attracting attention from both traditional medicine and modern science, possessing diverse pharmacological potential.

[0061] In the present invention, the *Hallyeocho* plant may be one or more selected from the group consisting of leaves, stems, fruits, roots, and flowers, and may be, for example, leaves, but is not limited thereto.

[0062] The term "Magnolia officinalis" in this specification refers to a deciduous tree belonging to the Magnoliaceae family, a medicinal plant native to East Asia, particularly China, Korea, and Japan. Magnolia officinalis is characterized by large flowers and thick bark, and has a long history and tradition as a medicinal plant. This tree typically grows to a height of 10 to 15 meters and has thick, sturdy trunks reaching a diameter of 30 to 50 cm. The leaves are elliptical or oblong, measuring 10 to 20 cm in length and 5 to 10 cm in width; they are glossy dark green and have slightly wavy margins.

[0063] The flowers of the Machilus thunbergii bloom in spring; they are large, white or light pink flowers that emit a slightly fragrant scent. In summer, round fruits form where the flowers have withered, containing numerous seeds. The Machilus thunbergii grows wild in mountainous regions and prefers fertile, well-drained soil. It grows quickly, and propagation is easy as branches frequently sprout from the roots.

[0064] The part of Magnolia bark used as a medicinal ingredient is primarily the bark. Magnolia bark has a thick, tough surface and emits a distinctive fragrance, playing an important role as a medicinal material. In Korean traditional medicine, Magnolia is mainly utilized for digestive system disorders and is effective in alleviating symptoms such as gastrointestinal discomfort, indigestion, and abdominal bloating. Magnolol and honokiol, the main pharmacological components of Magnolia, promote digestion and contribute to soothing the stomach and intestines through anti-inflammatory, antibacterial, and sedative effects.

[0065] Magnolia bark helps relieve stress and reduce anxiety, and can act as a nerve tranquilizer. It is also used to treat respiratory conditions such as coughs, phlegm, and asthma, and these efficacyes have been confirmed in both traditional medicine and modern research. Magnolia bark has been found to possess antioxidant and anti-inflammatory properties, and due to its sedative and antipyretic effects, it is used to alleviate symptoms caused by high fever.

[0066] Due to these diverse pharmacological effects, Magnolia bark has established itself as a valuable medicinal plant utilized for various medical purposes in both the East and the West.

[0067] In the present invention, the Magnolia bark may be one or more selected from the group consisting of leaves, stems, fruits, roots, seeds, and flowers, for example, it may be a stem, and specifically, it may be the bark of a stem, but is not limited thereto.

[0068] Another example of the present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, comprising attractylodin, wedelolactone, and / or honokiol as active ingredients.

[0069] In this invention, atractylodin is known to inhibit the production of pro-inflammatory mediators PGE2 and nitric oxide (NO) by suppressing the inflammatory cytokine IL-6. Accordingly, this invention confirmed that atractylodin has the effect of regulating inflammatory responses in the brain and inhibiting the expression of Lipocalin-2 (LCN2), a factor related to neuroregeneration. LCN2 is known to be elevated in patients with mild cognitive impairment and Alzheimer's disease, and recent studies have confirmed that LCN2 can inhibit neurogenesis through binding to the SLC22A17 receptor. Through this study, it was revealed that atractylodin can effectively regulate such expression.

[0070] In addition, it was found that atractylodin inhibits the reactivity of astrocytes and microglia that are abnormally activated due to iron excess and oxidative stress, prevents neuronal damage by reducing the expression of inflammatory cytokines (IL-1β, TNF-α), and alleviates the inhibitory effect on neurogenesis caused by inflammatory responses and oxidative stress. Through this, the present invention has revealed that atractylodin can contribute to neuronal protection and generation, as well as the maintenance of brain cognitive function.

[0071] In the present invention, the structural formula of atractylodine is as shown in the following structural formula 1.

[0072] [Structural Formula 1]

[0073]

[0074] In the present invention, wedelolactone has the effect of mitigating iron metabolism and oxidative stress responses by regulating the Keap1 / Nrf2 pathway. In the present invention, it was confirmed that wedelolactone increases the activity of Nrf2 by inhibiting the activation of Keap1, and thereby promotes the expression of Ferritin and HMOX1, which has the effect of reducing free iron accumulation and ROS production. In addition, wedelolactone was shown to reduce the expression of LCN2 / SLC22A17 by inhibiting the overactivation of the NF-κB and STAT3 pathways, thereby mitigating iron metabolism disturbances and inhibiting neuronal damage and inflammatory responses.

[0075] Furthermore, wedelolactone showed the effect of alleviating oxidative damage and neuroinflammation through the Fenton reaction and preventing the vicious cycle of tissue damage by blocking the overactivation of the LCN2 / SLC22A17 pathway and inhibiting free iron accumulation and ROS generation. Through this, the present invention revealed that wedelolactone can perform iron metabolism regulation and neuroprotective effects.

[0076] In the present invention, the structural formula of wedelolactone is as shown in structural formula 2 below.

[0077] [Structural Formula 2]

[0078]

[0079] In the present invention, honokiol exhibits a neuroprotective effect by protecting mitochondrial function, alleviating energy metabolism disorders, and suppressing the overactivity of astrocytes and microglia. Accordingly, the present invention has confirmed that honokiol has the effect of preventing a decrease in ATP production of glial cells (astrocytes and microglia) and promoting the normal differentiation and proliferation of neural progenitor cells.

[0080] Furthermore, honokiol was shown to prevent energy metabolism problems in astrocytes by reducing OxPhos (oxidative phosphorylation) and activating glycolysis, thereby decreasing the amount of lipid droplets accumulated in astrocytes due to lipid metabolism issues, and by regulating the acetylation and activation of STAT3 through the inhibition of excessive acetyl-CoA accumulation.

[0081] Furthermore, honokiol maintains the function of ABCA1 to facilitate cholesterol delivery from astrocytes to neurons, thereby playing a role in preventing neuronal developmental and functional decline, and promotes the normalization of glial cells through interaction with the two aforementioned substances.

[0082] In the present invention, the structural formula of honokiol is as shown in structural formula 3 below.

[0083] [Structural Formula 3]

[0084]

[0085] In the present invention, it was determined that the combination of the three substances above regulates energy metabolism and exhibits an effect that contributes to neuronal protection and functional recovery. Specifically, it was confirmed that the reduction of AMPK / PGC-1α is restored to normal levels, and the reduction of NGFR (NGF Receptor) is alleviated through the regulation of the activity of PFKP, a key enzyme in glycolysis.

[0086] Through this, it was confirmed that the expression of BDNF, which was reduced as NGFR was activated, increased, and the functions of NPY (Neuropeptide Y), ABCA (ATP-binding cassette transporter A), etc. were restored in turn.

[0087] In addition, the combination of the three substances was shown to alleviate the neuroinflammatory response induced by LCN2 and the inhibitory effects on neurogenesis and survival by suppressing the expression of LCN2 / SLC22A17. Although LCN2 / SLC22A17 can interfere with the mechanism of neuronal generation by inhibiting the NGFR signaling pathway, the combination of the three substances of the present invention reduces the expression of LCN2 / SLC22A17 and thereby plays a role in regulating NGFR signaling to function normally.

[0088] Therefore, the present invention has identified that a combination of three substances has the effect of restoring the function of NGFR, regulating energy metabolism, and suppressing neuroinflammation and neural circuit damage induced by LCN2.

[0089] In the present invention, brain cognitive function-related diseases may be one or more selected from the group consisting of neurodegenerative diseases, mental health and mood disorders, cognitive impairment and dementia, developmental disorders, nerve damage and trauma, Creutzfeldt-Jakob disease, chronic traumatic encephalopathy (CTE), and epilepsy, but are not limited thereto.

[0090] In the present invention, neurodegenerative diseases may be one or more selected from the group consisting of Alzheimer's disease, Parkinson's disease, Dementia with Lewy Bodies, Huntington's disease, Amyotrophic Lateral Sclerosis (ALS), Multiple Sclerosis (MS), Multiple System Atrophy (MSA), and Progressive Supranuclear Palsy (PSP), but are not limited thereto.

[0091] In the present invention, mental health and mood disorders may be one or more selected from the group consisting of depression, bipolar disorder, schizophrenia, generalized anxiety disorder (GAD), obsessive-compulsive disorder (OCD), and post-traumatic stress disorder (PTSD), but are not limited thereto.

[0092] In the present invention, cognitive impairment and dementia may be one or more selected from the group consisting of vascular dementia, frontotemporal dementia, mild cognitive impairment (MCI), Creutzfeldt-Jakob disease (CJD), and senile dementia, but are not limited thereto.

[0093] In the present invention, developmental disability may be one or more selected from the group consisting of Attention Deficit Hyperactivity Disorder (ADHD), Autism Spectrum Disorder (ASD), Learning Disabilities, and Intellectual Disability, but is not limited thereto.

[0094] In the present invention, nerve damage and trauma may be one or more selected from the group consisting of traumatic brain injury (TBI), stroke, chronic traumatic encephalopathy (CTE), and hypoxic brain injury, but are not limited thereto.

[0095] The pharmaceutical composition of the present invention may contain pharmaceutically effective amounts of extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark.

[0096] The pharmaceutical composition of the present invention may comprise a pharmaceutically effective amount of attractilodine, wedelolactone and / or honokiol.

[0097] The composition of the present invention may further comprise various auxiliary agents, such as pharmaceutically suitable and physiologically acceptable carriers, excipients, diluents, preservatives, stabilizers, flavoring agents, and flavoring agents, in addition to extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark; or atractylodine, wedelolactone, and / or honokiol. Furthermore, the present invention may appropriately select such auxiliary agents so as to interact with the active ingredients of the composition to improve the stability, absorption rate, bioavailability, etc., of the composition.

[0098] Carriers, excipients, and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0099] When formulating, commonly used diluents or excipients such as fillers, fillers, binders, wetting agents, disintegrants, and surfactants may be used. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and these solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin, with the extract. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used, which contribute to improving the flowability of the formulation and enhancing the mechanical properties of tablets or capsules. Additionally, substances such as disintegrants (e.g., sodium croscarmellose, crospovidone) may be included to improve the disintegration of the solid dosage form.

[0100] Preparations for oral administration include suspensions, liquid formulations, emulsions, syrups, and ointments. In addition to commonly used simple diluents such as water and liquid paraffin, these preparations may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. For example, polyethylene glycol (PEG) may be used as a humectant, and sucrose, fructose, and artificial sweeteners (e.g., aspartame, sucralose) may be used as sweeteners. Additionally, menthol and zinc compounds may be utilized as flavorings, and parabens, sodium benzoate, and potassium sorbate may be used as preservatives to enhance product stability and contribute to inhibiting microbial growth.

[0101] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and dermal preparations. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol (PEG), vegetable oils such as olive oil, injectable esters such as ethyl oleate, mineral oil, medium-chain triglycerides (MCT oil), and argan oil. Additionally, in injectable preparations, benzyl alcohol, chlorobutanol, polysorbate, and stearic acid may be used as additives, which can contribute to improving the stability, solubility, and bioavailability of the preparation.

[0102] Witepsol, Macrogol, Tween 61, cacao oil, lauroyl, glycerogelatin, hydrogel, and cocoa butter may be used as formulations for suppositories. These substrates play an important role in maintaining the shape of the suppository and controlling the release of the drug. In addition, some suppositories may include stabilizers or flavoring agents in addition to these substrates, which can contribute to improving the stability of the formulation and patient acceptance.

[0103] The pharmaceutical composition according to the present invention may be administered to mammals, including humans, via various routes. The method of administration may be any commonly used method, such as oral, skin, intravenous, intramuscular, subcutaneous, ophthalmic, or inhalation. In particular, oral administration may be preferred, and methods such as topical administration through the skin or intravenous, intramuscular, or subcutaneous injection may also be considered. Each method of administration may be selected according to the characteristics of the drug and may contribute to optimizing the effective delivery and in vivo absorption rate of the drug.

[0104] For example, the composition of the present invention may be administered orally, intraorally, or sublingually in the form of tablets containing starch or lactose, or as capsules containing excipients or alone, or as elixirs or suspensions containing chemicals for flavoring or coloring. These liquid formulations may be formulated with pharmaceutically acceptable additives such as glyceride mixtures (e.g., methylcellulose, semi-synthetic glycerides such as Witepsol, mixtures of apricot kernel oil and PEG-6 esters, or mixtures of PEG-8 and caprylic / capric glycerides). Additionally, the formulation may further include various additives (e.g., preservatives, flavoring agents, flavoring agents, etc.) to improve stability, absorption rate, and bioavailability, which may contribute to improving the quality of the formulation and increasing patient acceptance.

[0105] The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on various factors, such as the formulation method, mode of administration, patient's age, body weight, gender, pathological condition, food intake, time of administration, route of administration, excretion rate, and response sensitivity. The appropriate dosage varies according to these factors, and typically, a skilled physician can easily determine and prescribe an effective dosage suitable for the patient's condition and the purpose of treatment or prevention. Furthermore, the dosage may be adjusted stepwise as needed, and it is important to establish the optimal dosage by monitoring the patient's response during the treatment period.

[0106] In specific examples of applying the composition of the present invention to humans, the composition of the present invention may be administered alone, but may also be administered mixed with a pharmaceutical carrier selected in consideration of the method of administration and standard pharmaceutical practice. Such a carrier may include a substance suitable for oral, injectable, or topical formulations, and may be, for example, an excipient, a lubricant, a preservative, a stabilizer, etc. Furthermore, the pharmaceutical carrier may play a role in improving the bioavailability of the drug, maintaining the stability of the drug, and improving patient acceptance.

[0107] The dosage of the extract-containing composition of the present invention may vary depending on the patient's age, body weight, gender, form of administration, health status, and the severity of the disease. Additionally, the dosage may be adjusted according to the purpose of treatment and the patient's response, and may be administered in divided doses from once to several times a day at regular intervals at the discretion of a physician or pharmacist. In this case, the appropriate dosage can be gradually optimized while monitoring the patient's condition and treatment response. Furthermore, for specific treatments or prevention, high or low dose administration may be necessary, and such adjustments should be made in accordance with the guidance of a medical professional.

[0108] In the present invention, the pharmaceutical composition may be administered by adjusting the content of extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark to a range from 5 mg / day to 500 mg / day per kg of body weight. For example, it may be set to a range such as 5 mg / day to 500 mg / day, 10 mg / day to 400 mg / day, 20 mg / day to 300 mg / day, 50 mg / day to 200 mg / day, etc., and may usually be administered in the range of 50 mg / day to 100 mg / day. These dosages may be adjusted according to the patient's condition, treatment purpose, and response.

[0109] If the daily dosage of the composition containing extracts of *Changchang*, *Hallyeocho*, and *Houbak* of the present invention is less than the above dosage, a significant effect cannot be obtained; and if it exceeds the above dosage, it is not only uneconomical but also exceeds the commercial dosage range, so there is a risk of undesirable side effects. Therefore, it is preferable to administer within the above range.

[0110] In the present invention, the pharmaceutical composition may be administered with the content of atractilodine, wedelolactone, and honokiol adjusted to a range from 5 mg / day to 500 mg / day per kg of body weight. For example, it may be set to a range such as 5 mg / day to 500 mg / day, 10 mg / day to 400 mg / day, 20 mg / day to 300 mg / day, 50 mg / day to 200 mg / day, etc., and may usually be administered in the range of 50 mg / day to 100 mg / day. These dosages may be adjusted according to the patient's condition, treatment purpose, and response.

[0111] If the daily dosage of the composition containing atractylodin, wedelolactone, and honokiol of the present invention is less than the above dosage, a significant effect cannot be obtained; and if it exceeds the above dosage, it is not only uneconomical but also outside the range of commercial dosage, so there is a risk of undesirable side effects. Therefore, it is preferable to administer within the above range.

[0112] In the present invention, the extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis may be crude extracts obtained by extraction with a polar solvent, a non-polar solvent, or a mixture thereof. Polar solvents may include water, alcohol, acetic acid, dimethyl formamide (DMF), dimethyl sulfoxide (DMSO), etc., and non-polar solvents may include hexane, chloroform, benzene, etc. These solvents may be selected according to the characteristics of the extract, but are not limited thereto.

[0113] In the present invention, the alcohol may be a straight-chain or branched alcohol having 1 to 4 carbon atoms, for example, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol, ethylene glycol, or a mixture thereof, but is not limited thereto. Such alcohol may be used as a polar solvent in the extraction and dissolution process of the extract of the present invention, and specific alcohols may be selected according to solubility and extraction efficiency. In addition, the type of alcohol that can be used in the present invention may vary depending on the characteristics of the substance to be extracted.

[0114] In the present invention, when the solvent is a mixture of water and alcohol, the alcohol concentration of the mixture may be in the range of 10% or more to less than 100% (v / v), 20% or more to less than 100% (v / v), 30% or more to less than 100% (v / v), 10% or more to 90% (v / v), 20% or more to 90% (v / v), 30% or more to 90% (v / v), 10% or more to 80% (v / v), 20% or more to 80% (v / v), 30% or more to 80% (v / v), 10% or more to 70% (v / v), 20% or more to 70% (v / v), and 30% or more to 70% (v / v). For example, an aqueous solution of a straight-chain or branched alcohol having 1 to 4 carbon atoms at a concentration of 30% (v / v) This may be possible. Such an alcohol concentration range can be optimized according to extraction efficiency and solubility, and the alcohol concentration that can be used in the present invention is not limited thereto. In addition, the concentration and type of alcohol may vary depending on the properties and uses of the extract.

[0115] In the present invention, the fractions may consist of various fractions obtained by sequentially fractionating a crude extract obtained by solvent extraction with solvents such as normal hexane, chloroform, ethyl acetate, and water-saturated butanol; for example, they may include a normal hexane fraction, a chloroform fraction, an ethyl acetate fraction, and a water-saturated butanol fraction. Additionally, crude extracts obtained using hydrophilic or non-hydrophilic solvents such as water and ethanol may also be included. Water extraction is primarily used to extract water-soluble components, while ethanol extraction can extract both water-soluble and water-insoluble components, enabling the extraction of various compounds. Furthermore, high-efficiency extraction methods such as supercritical extracts may also be used. These extracts may exhibit physiological effects depending on their different physicochemical properties, and a specific fraction may contain the most effective active ingredient. In the present invention, mixtures or individual fractions of these fractions may be suitably used, but are not limited thereto.

[0116] In the present invention, the extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis may be mixed in a volume ratio of 1 to 8:1 to 8:1 to 8, for example, in a volume ratio of 4.5:4.5:1, but are not limited thereto.

[0117] In the present invention, the concentration of the extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis may be 100 to 400 mg / kg, for example, 200 mg / kg, but is not limited thereto.

[0118] The pharmaceutical composition of the present invention may comprise pharmaceutically effective amounts of attractilodine, wedelolactone, and honokiol.

[0119] Another example of the present invention relates to a food composition for improving brain cognitive function comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0120] Another example of the present invention relates to a food composition for improving brain cognitive function comprising atractylodin, wedelolactone, and honokiol as active ingredients.

[0121] In the present invention, there are no special limitations on the types of food. Examples of foods to which the above composition can be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, diet foods, special purpose foods (e.g., immunity boosting, digestion promotion, etc.), and include all foods in the conventional sense.

[0122] In the present invention, the beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, as well as natural sweeteners such as steviol glycosides and monk fruit extract, or synthetic sweeteners such as saccharin, aspartame, acesulfame potassium, and sucralose. Additionally, ingredients such as dietary fiber, vitamins, minerals, amino acids, and lactic acid bacteria may be added to enhance the functionality of the beverage. The ratio of the natural carbohydrates and other added ingredients may be appropriately determined by the choice of a person skilled in the art.

[0123] In addition to the above, the food composition of the present invention may contain various nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, it may include functional ingredients such as sweeteners (e.g., natural sweeteners and synthetic sweeteners), emulsifiers, probiotics, prebiotics, polyphenols, and flavonoids. In addition, the food composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination, and the proportion of the corresponding additives may also be appropriately selected by those skilled in the art.

[0124] When the food composition of the present invention is used as a food additive, the food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. Generally, when manufacturing food or beverages, the food composition of the present invention may be added in a range of 0.01 to 5 weight% relative to the raw material, preferably in an amount of 0.05 to 3 weight%, and more preferably in an amount of 0.1 to 2 weight%.

[0125] In the food composition of the present invention, details that overlap with the pharmaceutical composition are omitted for the sake of complexity of this specification.

[0126] Another example of the present invention relates to a method for preparing a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0127] The term "extract" as used in this specification refers to a crude extract as commonly used in the art, but in a broader sense, it also includes a fraction that has undergone an additional fractionation process. That is, the extract of the present invention includes not only a primary extract obtained using a specific extraction solvent, but also a fraction obtained by additionally applying a purification process thereto.

[0128] For example, 20g of each ground sample is weighed, and 200mL of ethanol (EtOH) at concentrations of 10%, 30%, and 70% is added to each sample, and the extract is extracted by water bath (Digital General Purpose Water Bath, DAIHAN Scientific Co., Ltd., Seoul, Republic of Korea) at 65℃ for 2 hours. The extracted solution is filtered, concentrated using a vacuum concentrator (Rotary Evaporator N-Series, EYELA, Tokyo Rikakikai Co., Ltd., Tokyo, Japan), and freeze-dried at -70℃ to produce a powder-form extract, which is then stored frozen at -20℃.

[0129] The extracted substance may undergo additional purification processes, for example, including fractionation using an ultrafiltration membrane having a specific molecular weight cutoff value, and separation processes by various chromatographs (e.g., size exclusion, ion exchange, hydrophobic interaction, affinity chromatography, etc.). Fractions obtained through such additional purification methods are also included in the extract of the present invention. Furthermore, the extract of the present invention may be provided in the form of a solution, concentrate, or powder.

[0130] The extraction method used in the present invention may include various methods that can be commonly used. For example, maceration, hot water extraction, ultrasonic extraction, reflux extraction, vacuum extraction, microwave-assisted extraction, etc. may be used, but are not limited thereto.

[0131] The process of preparing extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark according to the present invention is described in more detail as follows:

[0132] First, a certain amount of powders of Atractylodes macrocephala, Eclipta prostrata, and Magnolia officinalis is prepared, and an extraction solvent of about 8 to 50 times (by weight) is added to them to perform extraction at room temperature or by heating. After extraction, the filtrate is recovered by filtration. There are no specific limitations on the extraction temperature, but a range of 15 to 110°C, preferably 20 to 90°C, is appropriate.

[0133] The extraction process may be repeated once or several times, and in one preferred embodiment of the present invention, a method of performing re-extraction after the first extraction may be adopted. This is because it is difficult to obtain sufficient extraction efficiency with only the first extraction during mass production, and losses may occur during the extraction process in the case of raw materials with high inherent moisture content. As a result of verifying the extraction efficiency at each stage, it was found that approximately 80 to 90% of the total active ingredients are extracted when a second extraction is performed.

[0134] In the present invention, reflux extraction can be performed preferably twice, and more preferably 8 to 12 times by volume. After extraction, the mixture is filtered, and when repeated extraction is performed in the same manner, an extraction solvent (about 5 to 15 volume ratio) is added again to the obtained residue, combined with the filtrate, and then concentrated under reduced pressure to produce extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark. Although extraction efficiency can be maximized by performing two extractions in this manner and mixing the filtrates after each extraction, the extracts of the present invention are not necessarily limited to a specific number of extractions.

[0135] When preparing extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis, using too little extraction solvent makes stirring difficult and may reduce extraction efficiency due to low solubility of active ingredients. Conversely, using an excessive amount of solvent may necessitate excessive solvent removal during the subsequent purification process, which can lower economic efficiency and make handling difficult. Therefore, it is desirable to maintain an appropriate amount of solvent.

[0136] The filtered extract, after extraction is complete, can have the content of residual lower alcohols controlled to make it suitable for use as a raw material for pharmaceuticals and food. To this end, water of about 10 to 30 times (preferably 15 to 25 times, more preferably about 20 times) the total amount of concentrate is added and azeotropically concentrated 1 to 5 times (preferably 2 to 3 times), then an equal amount of water is added again to homogeneously suspend the mixture, and then freeze-dried to produce powdered extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis.

[0137] Another example of the present invention relates to a method for preparing a composition comprising attractylodin, wedelolactone and / or honokiol as active ingredients.

[0138] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0139] Another example of the present invention is to provide a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for the prevention, improvement, or treatment of brain cognitive function-related diseases.

[0140] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0141] Another example of the present invention relates to the use of a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for improving brain cognitive function.

[0142] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0143] Another example of the present invention relates to a method for preventing, improving, or treating brain cognitive function-related diseases using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0144] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0145] Another example of the present invention relates to a method for improving brain cognitive function using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0146] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0147] Another example of the present invention is to provide a composition comprising attractilodine, wedelolactone, and honokiol as active ingredients for the prevention, improvement, or treatment of brain cognitive function-related diseases.

[0148] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0149] Another example of the present invention relates to the use of a composition comprising atractylodin, wedelolactone, and honokiol as active ingredients for improving brain cognitive function.

[0150] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0151] Another example of the present invention relates to a method for preventing, improving, or treating brain cognitive function-related diseases using a composition comprising atractylodin, wedelolactone, and honokiol as active ingredients.

[0152] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0153] Another example of the present invention relates to a method for improving brain cognitive function using a composition comprising atractylodin, wedelolactone, and honokiol as active ingredients.

[0154] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0155] Another example of the present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases in companion animals, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0156] In the present invention, brain cognitive function-related diseases of companion animals may be Cognitive Dysfunction Syndrome (CDS), Canine Dementia (Feline Dementia), Cerebrovascular Disease, or Meningoencephalitis, but are not limited thereto.

[0157] In specific examples of applying the composition of the present invention to companion animals, the composition of the present invention may be administered alone, but may also be administered mixed with a veterinary carrier selected in consideration of the method of administration and veterinary standard practice. Such a carrier may include a substance suitable for oral, injectable, or topical formulations, and may be, for example, an excipient, a lubricant, a preservative, a stabilizer, etc. Furthermore, the veterinary carrier may play a role in improving the bioavailability of the drug, maintaining stability, and improving palatability so that the companion animal can ingest it more easily.

[0158] The dosage of the extract-containing composition of the present invention may vary depending on the age, weight, breed, form of administration, health status, and the degree of progression of cognitive impairment syndrome of the pet. Additionally, the dosage may be adjusted according to the therapeutic purpose and the pet's response, and may be administered in divided doses from once to several times a day at regular intervals at the discretion of a veterinarian. In this case, the appropriate dosage can be gradually optimized while monitoring the pet's condition and therapeutic response. Furthermore, high or low dose administration may be necessary for specific therapeutic or preventive purposes, and such adjustments should be made in accordance with veterinary guidelines and expert recommendations.

[0159] In the present invention, the pharmaceutical composition for companion animals may be administered by adjusting the content of extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark to a range from 5 mg / day to 500 mg / day per kg of the animal's body weight. For example, for dogs, the standard dose for small dogs weighing 10 kg or less is one packet per day based on a core ingredient purity of 20%; for medium dogs (over 10 kg to 20 kg or less), it may be 1.5 times the standard dose for small dogs; for large dogs (over 20 kg to 40 kg), it may be 2 times the standard dose for small dogs; and for extra-large dogs (40 kg or more), it may be 3 times the standard dose for small dogs. For cats, the dosage may generally be adjusted by considering a ratio half that of dogs weighing 10 kg or less. These dosages may be adjusted according to the animal's condition, the progression of the disease, the purpose of treatment, and the response; it is preferable to determine the optimal dosage based on the judgment and guidelines of a veterinarian.

[0160] If the daily dosage of the composition containing extracts of *Changchang*, *Hallyeocho*, and *Houbak* of the present invention is less than the above dosage, a significant effect cannot be obtained; and if it exceeds the above dosage, it is not only uneconomical but also exceeds the commercial dosage range, so there is a risk of undesirable side effects. Therefore, it is preferable to administer within the above range.

[0161] To avoid complexity in the specification, descriptions of parts identical to those described above are omitted.

[0162] Another example of the present invention relates to a feed composition for improving brain cognitive function comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0163] As used in this specification, the term "feed composition" refers to food fed to animals. The said feed composition refers to a substance that supplies organic or inorganic nutrients necessary for maintaining the life of an animal or for producing meat, milk, etc. The said feed composition may additionally include nutritional components necessary for maintaining the life of an animal or for producing meat, milk, etc.

[0164] In the present invention, the feed composition may be prepared in the form of powder or granules, and may additionally include one or more of the following: organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid; phosphates such as sodium phosphate, potassium phosphate, acidic pyrophosphate, and polyphosphate; and natural antioxidants such as polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid, but is not limited thereto.

[0165] In the present invention, the feed composition may further include grains, for example, ground or crushed wheat, oats, barley, corn, and rice; plant protein feed, for example, feed with rapeseed, soybeans, and sunflower as main components; animal protein feed, for example, blood meal, meat meal, bone meal, and fish meal; and dried components consisting of sugars and dairy products, for example, various milk powders and whey powders, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, etc., but is not limited thereto.

[0166] In the present invention, the feed composition may be administered to animals alone or in combination with other feed additives among edible carriers. Additionally, the feed composition may be easily administered to animals by directly mixing it with feed or as an oral formulation separate from the feed.

[0167] When the above feed composition is administered separately from the feed, it may be prepared into an immediate-release or sustained-release formulation by combining it with a feed-grade edible carrier, as is well known in the art. Such edible carriers may be solid or liquid, for example, corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When a solid carrier is used, the feed composition may be a tablet, capsule, powder, troche, sugar-containing tablet, or a finely dispersed top dressing. When a liquid carrier is used, the feed composition may be in the form of a gelatin soft capsule, syrup, suspension, emulsion, or solution, but is not limited thereto.

[0168] In the present invention, the feed composition may contain, for example, preservatives, stabilizers, wetting agents or emulsifiers, solution accelerators, etc., but is not limited thereto.

[0169] In the present invention, the feed composition may be used by adding it to animal feed by soaking, spraying, or mixing, but is not limited thereto.

[0170] In the present invention, the feed composition can be applied to the diets of a number of animals, including mammals and poultry. The mammals may be used for pigs, cattle, sheep, goats, laboratory rodents, and other animals, as well as companion animals, such as dogs and cats, and the poultry may be used for chickens, turkeys, ducks, geese, pheasants, and quail, but are not limited thereto.

[0171] In the present invention, the feed composition can be formulated in the form of a conventional feed and may include conventional feed ingredients.

[0172] In the present invention, the feed composition is designed to maximize the improvement of cognitive function and neuroprotective effects in companion animals through a combination of auxiliary components.

[0173] Accordingly, the feed composition of the present invention may additionally include auxiliary components that perform various physiological roles for maintaining cognitive function and neuroprotecting companion animals.

[0174] In the present invention, the auxiliary component may be citric acid. Citric acid promotes intracellular metabolism and plays a role in the process of energy generation in the body, thereby helping nerve cells function smoothly.

[0175] In addition, the auxiliary component in the present invention may be zeolite. Zeolite acts as a stabilizer and viscosity modifier to maintain the homogeneity of the composition and improve the bioavailability of the active ingredient.

[0176] In the feed composition of the present invention, details that overlap with the pharmaceutical composition are omitted for the sake of complexity of this specification.

[0177] Another example of the present invention is to provide a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for the prevention, improvement, or treatment of brain cognitive function-related diseases.

[0178] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0179] Another example of the present invention relates to the use of a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients for improving brain cognitive function.

[0180] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0181] Another example of the present invention relates to a method for preventing, improving, or treating brain cognitive function-related diseases using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0182] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0183] Another example of the present invention relates to a method for improving brain cognitive function using a composition comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0184] Regarding this, to avoid complexity in the specification, the description of parts identical to those described above is omitted.

[0185] The present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same. Specifically, the invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark as active ingredients; and a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising Atractylodin, Wedelolactone, and / or Honokiol as active ingredients.

[0186] FIG. 1a is a graph showing the results of a safety evaluation in 10% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0187] FIG. 1b is a graph showing the results of a safety evaluation in 30% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0188] FIG. 1c is a graph showing the results of a safety evaluation in 70% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0189] FIG. 2a is a graph showing the results of evaluating the anti-inflammatory efficacy of 10% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0190] FIG. 2b is a graph showing the results of evaluating the anti-inflammatory efficacy of 30% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0191] FIG. 2c is a graph showing the results of evaluating the anti-inflammatory efficacy of 70% EtOH extracts of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis, according to one embodiment of the present invention.

[0192] FIG. 3a is a graph showing the results of the cytotoxicity evaluation of a mixture of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark according to one embodiment of the present invention.

[0193] FIG. 3b is a graph showing the results of evaluating the anti-inflammatory efficacy of a mixture of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark according to one embodiment of the present invention.

[0194] Figure 4 is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark, and a single extract, according to one embodiment of the present invention.

[0195] Figure 5 is a graph showing the results of an evaluation of the memory-improving efficacy of three types of mixtures of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark, according to one embodiment of the present invention.

[0196] FIG. 6a is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes japonica, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in a scopolamine-induced amnesia model according to one embodiment of the present invention.

[0197] FIG. 6b is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes macrocephala, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in a Scopolamine-induced amnesia model according to one embodiment of the present invention.

[0198] FIG. 6c is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes japonica, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in a Scopolamine-induced amnesia model according to one embodiment of the present invention.

[0199] FIG. 7a is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes japonica, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in an animal model of Amyloid beta-induced Alzheimer's disease according to one embodiment of the present invention.

[0200] FIG. 7b is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes japonica, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in an animal model of Amyloid beta-induced Alzheimer's disease according to one embodiment of the present invention.

[0201] FIG. 7c is a graph showing the results of evaluating the memory-improving efficacy of a mixture of Atractylodes japonica, Erythronium japonicum, and Magnolia officinalis (4.5:4.5:1) in an animal model of Amyloid beta-induced Alzheimer's disease according to one embodiment of the present invention.

[0202] A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

[0203] The present invention will be explained in more detail below through the following examples. However, these examples are merely illustrative of the invention, and the scope of the invention is not limited by these examples.

[0204] Throughout this specification, "%" used to indicate the concentration of a particular substance is (weight / weight) % for solid / solid, (weight / volume) % for solid / liquid, and (volume / volume) % for liquid / liquid, unless otherwise noted.

[0205]

[0206] Example 1. Preparation of a natural complex for cognitive improvement

[0207] 1-1. Preparation of Natural Product Extracts

[0208] The samples listed in Table 1 below were ground, weighed at a rate of 20g each, and 200mL of 10%, 30%, and 70% EtOH were added, respectively. Extraction was performed by heating in a water bath (Digital General Purpose Water Bath, DAIHAN Scientific Co., Ltd., Seoul, Republic of Korea, Cat. No. DH.WB000106) at 65℃ for 2 hours. Subsequently, the extracts were filtered and concentrated using a vacuum concentrator (Rotary Evaporator N-Series, EYELA (Tokyo Rikakikai Co., Ltd.), Tokyo, Japan, Cat. NO. 243820). Then, the extracts were freeze-dried at -70℃ (Bench Top Freeze Dryer, For Laboratory, OPERON, Gyeonggi-do, Republic of Korea, Cat. No. FDB-5503) to produce a powder-form extract, which was then stored frozen at -20℃.

[0209] In addition, the yield in a completely powdered state after freeze-drying was measured using the following formula 1 and is shown in Table 1.

[0210] [Formula 1]

[0211] Yield (%) = (Weight of extract / Weight of initial sample) x 100

[0212] Classification Scientific Name Yield (%) 10% EtOH 30% EtOH 70% EtOH Atractylodes lancea DC 28.60 21.30 7.95 Atractylodes chinensis Koidzumi 22.50 13.25 13.45 Eclipta prostrata Linne 22.15 22.25 16.80 Machilus thunbergii Siebold & Zucc. 5.45 7.20 5.25

[0213]

[0214] 1-2. Preparation of Extracts and Formulations

[0215] Considering the difficulty of weighing each extract and mixture at the microscopic level, the stock solution prepared at a 100-fold concentration was diluted at a ratio of 1:99 in FBS-free Dulbecco's Modified Eagle's Medium (1X), liquid (With 4500 mg / L D-glucose, L-glutamine)) (DMEM) for use. Single samples of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis were weighed according to their respective concentrations, dissolved in a vehicle, and prepared by vortexing followed by a 5-minute sonication process (POWER SONIC410, Gyeonggi-do, Republic of Korea). Mixture samples were weighed according to the ratio of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis based on the final concentration, dissolved in a vehicle in the same manner, and used after vortexing followed by a 5-minute sonication process.

[0216]

[0217] Example 2. Experimental Method

[0218] 2-1. Cell Culture

[0219] Mouse-derived brain macrophages, BV2 microglia (Cytion, Cat #305156), were cultured in a medium prepared by adding 100 unit / mL penicillin-streptomycin (P / S) and 10% fetal bovine serum (FBS) to Dulbecco's Modified Eagle's Medium (1X), liquid (With 4500 mg / L D-glucose, L-glutamine) (DMEM). Cells were cultured in a CO2 incubator (Air Jacket, VISION SCIENTIFIC Co., Ltd, Daejeon-Si, Republic of Korea) at 37°C and 5% CO2 conditions and subcultured every 3 days.

[0220]

[0221] 2-2. Cytotoxicity Assessment

[0222] 2 x 10 BV2 microglia cells 5 Cells were seeded into a 96-well cell culture plate at cells / well and cultured in an incubator for 24 hours. Subsequently, the culture medium was replaced with FBS-free DMEM, and the cells were cultured in the incubator for another 24 hours. After 24 hours, the culture medium was removed, and 100 μL of medium containing the extract was added to each well. After exposing the cells to the DMEM containing the extract for 24 hours, 5 mg / mL MTT reagent was mixed with FBS-free DMEM at a 1:9 ratio, and the cells were cultured in the incubator for 4 hours. Subsequently, the supernatant was removed, and 100 μL of a mixture of 99.5% DMSO and iso-propanol in a 1:1 ratio was dispensed into each well to lyse the cells. Cell viability was calculated by measuring absorbance at 540 nm using a SpectraMax 190 Microplate reader (Molecular Devices, San Jose, CA, USA).

[0223]

[0224] 2-3. Evaluation of Anti-inflammatory Efficacy

[0225] 2 x 10 BV2 microglia cells 6Cells were seeded into a 6-well cell culture plate at a cell / well ratio and cultured in an incubator for 24 hours. Subsequently, the culture medium was replaced with FBS-free DMEM, and the cells were cultured in the incubator for another 24 hours. After 24 hours, the extract was added to the culture medium for pretreatment, and 1 hour later, LPS (1 μg / mL) was added to all wells except the control group to stimulate the cells for 24 hours to induce an inflammatory response. Afterward, the cell culture supernatant was mixed with Griess reagent in a 1:1 ratio and reacted; 200 μL of the mixture was dispensed into 96-well plates, and the absorbance was measured at 540 nm using a SpectraMax 190 Microplate reader (Molecular Devices, San Jose, CA, USA). NO concentration was quantified after constructing a standard curve using sodium nitric oxide (NaNO2).

[0226]

[0227] 2-4. Test Animals

[0228] Male ICR mice aged 5 weeks were obtained from Coatec Co., Ltd. (Pyeongtaek, Korea) and reared at the Laboratory Animal Center of Kangwon National University. The rearing environment was maintained at a temperature of 23 ± 2 ℃, humidity of 50 ± 10%, and a 12-hour light-dark cycle (06:00-18:00). During the study period, laboratory solid feed (2018S; Envigo, Madison, WI, USA) and water were provided free of charge via tap water. In addition, to minimize stress on the animals, all experiments were performed after an acclimatization period of at least one week following introduction to the water. This animal experiment was conducted with the approval of the Kangwon National University Animal Ethics Committee (KW-241104-1).

[0229]

[0230] 2-5. Scopolamine Induction and Test Substance Administration

[0231] Scopolamine is a muscarinic receptor antagonist used to induce short-term amnesia. 1 mg / kg of scopolamine was administered intraperitoneally (ip) 30 minutes prior to all behavioral tests, and the test substance was administered orally (po) 30 minutes prior to scopolamine administration to expose the experimental animals.

[0232]

[0233] 2-6. Amyloid beta injection and administration of test substance

[0234] Amyloid beta (Aβ 1-42, Bachem) was dissolved in hexafluoroisopropanol (HFIP) and incubated at room temperature for 2 hours to decompose into nanoparticles. Subsequently, the HFIP was completely evaporated to remove it, and the nanoparticle amyloid beta was dissolved in 1X PBS and incubated at 37°C for 48 hours to induce aggregation. Experimental animals were anesthetized with avertin, and with their bodies and heads immobilized, amyloid beta (20 μM, 3 μL) was slowly injected via intraventricular (ICV) injection. The control group was injected with the same dose of sterile 1X PBS under identical conditions, and the incision site was sutured. After acclimatizing the animals for one week to allow the amyloid beta to oligomerize within the brain, the test substance was administered orally (per oral, po) for one week. Subsequently, the behavioral study schedule was carried out.

[0235]

[0236] 2-7. Y-shaped Maze Test

[0237] A Y-shaped maze consisting of three branches was randomly designated as A, B, and C. The experimental animals were placed in the center of the maze and allowed to explore freely for 8 minutes, after which the branches the animals entered were recorded. Cases where animals entered three different branches in sequence were judged as actual alternation, and spontaneous alternation (%) was calculated using Formula 2 below.

[0238] [Formula 2]

[0239] Spontaneous alternation (%) = {Actual alternation / Total entry count - 2} × 100

[0240]

[0241] 2-8. Manual Avoidance Test

[0242] The experiment was conducted for two days. On the first day, during the acquisition trial, experimental animals were placed in a bright area, and after 10 seconds, a guillotine-shaped doorway was opened to measure the latency time it took for the animals to move into the dark area. Once the animals entered the dark area, they were trained by administering a 0.5 mA electric shock for 3 seconds. For the retention trial, 24 hours later, the animals were placed in a bright area in the same manner as the previous day, and after the guillotine doorway was opened after 10 seconds, the time it took for them to enter the dark area was measured up to a maximum of 300 seconds.

[0243]

[0244] Example 3. Derivation of Optimal Mixing Ratio

[0245] 3-1. Evaluation of the Efficacy of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in BV2 microglia cells

[0246] 3.1.1. Results of Safety Evaluation of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in BV2 microglia cells

[0247] To evaluate the safety of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis, cytotoxicity was assessed by performing an MTT assay in a BV2 microglia cell model. Cytotoxicity was defined as a decrease in cell viability to 0.7 or lower when the control group's viability was standardized to 1, and the results are shown in Figures 1a to 1c and Tables 2 to 4 (Explanation of figures: *** p<0.001, ** p<0.01, * p<0.05 vs CON).

[0248] GROUPO.DMeanStd. DeviationStd. Error of MeanCON0.9907160.9727621.0402121.02688710.032030.0143200.4468540.5547730.5813230.5114930.54710.073140.03271 Parent Creation 1000.8936740.9632070.9176780.8970140.9190.027850.012452000.9496180.9474030.8756880.8767460.9220.042120.01884500 0.8998570.918670.888880.91990.036880.01844Hallyeoncho1000.835350.9451870.9484610.945220.92070.048290.021592000.9117270.862760.9042550.83740.8790.030430.013615000.8787960.8388220.8493030.858395 0.85770.015010.006711Magnolia bark 1000.883160.906040.9788130.9366570.91940.038940.017412000.9669430.9385420.9551720.9452530.93610.036010.01615000.9804660.971770.9735230.9540150.96940.009840.004401

[0249] GROUPO.DMeanStd. DeviationStd. Error of MeanCON0.9618121.0483410.9688241.07387310.057050.0255100.3046780.4210670.563190.5140380.44110.10090 .04512More creation1000.9710391.0265660.9587250.9531220.97160.031820.014232000.9270871.0800130.6701521.0351550 .83730.25790.11535001.0163451.1352611.0157621.0009331.0420.062530.03127Hanryeoncho1000.8933341.0449180.5199761.0330410.90290.22270.099592001.0869581.0347861.0568290.5015570.94490.24860.11125000.5129310.491458 0.6647830.55640.094480.05455Magnolia bark 1001.1333591.1300041.0833011.1010491.0960.041820.01872001.1201740.9702330.5243721.0479380.96050.25270.1135000.1573550.3641310.182250.1541010.20440.089960.04023

[0250] GROUPO.DMeanStd. DeviationStd. Error of MeanCON1.0034910.9478790.9968371.04245210.034020.0152100.5684640.4156430.50 83840.4221370.47030.066160.02959More creation1000.9958081.0127170.970220.9403250.9745 0.029760.013312000.937561.02851.0095340.98250.99140.034410.015395000.9011390.8833310.9259560.9377530.90840.022730.01016Hallyeoncho1000.9901830.9296850.9439890. 9622160.95310.023880.010682000.9521870.9282060.956880.9746890.9450.024330.010885000.3927560.3748180.3555310.1937420.33720.081290.03636Hubak1000.446311.07 99331.0671720.6397310.75630.29760.13312000.0308280.0588590.0867610.0300880.046890.025650.011475000.0694990.0781460.061880.0611090.066460.0073910.003305

[0251] As a result, Atractylodes lanceolata, Erythronium japonicum, and Magnolia officinalis did not exhibit cytotoxicity in the 10% EtOH extract (Fig. 1a), while cytotoxicity was confirmed in the 30% EtOH extract at 500 μg / mL of Erythronium japonicum and 500 μg / mL of Magnolia officinalis, as the cell viability was lower than 0.7 (Fig. 1b). Additionally, strong toxicity was confirmed in the 70% EtOH extract at 500 μg / mL of Erythronium japonicum and 200 and 500 μg / mL of Magnolia officinalis, as the cell viability was lower than 0.7 (Fig. 1c).

[0252]

[0253] 3.1.2. Evaluation of Anti-inflammatory Efficacy of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in BV2 Microglia Cells

[0254] To evaluate the anti-inflammatory efficacy of Atractylodes rhizome, Eclipta prostrata, and Magnolia officinalis, a NO assay was performed in a BV2 microglia cell model to assess cytotoxicity, and the results are shown in Figures 2a to 2c (Description of drawings: *** p<0.001 vs CON; ### p<0.001 vs 0 (LPS alone)).

[0255] GROUPO.DStd. DeviationStd. Error of MeanCON2.8933334.080.83910.5933082.8666785.106671.5841.12MoChangCheol10060.1733359.40.54680.386720050.6666750.466670.14140.150063.0133364.693331.1880.84Hanryeoncho10049.6133349.77 3330.11310.0820035.1066734.533330.40540.286750025.9466726.440.34880.2467Hubak10044.1466745.120.68820.486720034.4133334.680.18860.133350024.6933324.626670.047140.03333

[0256] GROUPO.DStd. DeviationStd. Error of MeanCON2.8933332.7466670.10370.07333087.3866785.521.320.9333MoChangCheol10058.1733359.133330.67880.4820016.6666716.866670.14140.150010.4666710.026670.31110.22Hanryeoncho10035 34.933330.047140.0333420021.6933318.453332.2911.6250010.1333310.120.0094260.006665Hoobak10010.94667110.037710.026672003.924.160.16970.125006.927.3866670.330.2333

[0257] GROUPO.DStd. DeviationStd. Error of MeanCON2.62.480.084850.06084.8133386.253331.0180.72Mochangchul10035.8133336.3333330.36770.2620014.815.160.25460.185002.5066672.6933330.1320.09333Hallyeoncho10027.7733327.693330 .056570.0420013.7333314.106670.2640.18675005.4533335.4666670.0094290.006667Hubak1004.124.0933330.018860.013332005.8933336.20.21680.15335007.488.1866670.49970.3533

[0258] As a result, in the 10% EtOH extract, *Atractylodes macrocephala*, *Atractylodes japonica*, *Eclipta prostrata*, and *Magnolia officinalis* all significantly reduced the amount of NO produced increased by LPS. However, the inhibitory effect was not distinct, and in particular, *Atractylodes japonica* 100 μg / mL showed a tendency for NO production to actually increase compared to the group treated with LPS alone (Fig. 2a). In the 30% EtOH extract, *Atractylodes macrocephala*, *Atractylodes japonica*, *Eclipta prostrata*, and *Magnolia officinalis* all significantly reduced the amount of NO produced increased by LPS, and the degree of inhibition of NO production was more pronounced than in the 10% EtOH extract (Fig. 2b). In the 70% EtOH extract, *Atractylodes macrocephala*, *Eclipta prostrata*, and *Magnolia officinalis* all significantly reduced the amount of NO produced increased by LPS, and the degree of inhibition of NO production was more pronounced than in the 10% and 30% EtOH extracts (Fig. 2c).

[0259] Consequently, the 70% EtOH extract demonstrated the best anti-inflammatory efficacy, but it presented a limitation due to the strong cytotoxicity of *Hyundai-cho* and *Magnolia officinalis*. Furthermore, considering the lowest yield of the 70% EtOH extract, it was determined that the 30% EtOH extract would be most suitable for the study, as it exhibits low cytotoxicity, anti-inflammatory efficacy, and a consistent yield. Accordingly, subsequent research was conducted primarily using the 30% EtOH extract. Additionally, since *Magnolia officinalis* extract may exhibit strong cytotoxicity when used alone, it was combined with *Atractylodes lanceolata*, *Hyundai-cho*, and *Magnolia officinalis* to address this. As this combination has the potential to maximize anti-inflammatory efficacy through synergistic effects, experiments were conducted on the blend to mitigate the strong toxicity of *Magnolia officinalis* and verify the synergistic effects.

[0260]

[0261] 3.1.3. Evaluation of Cytotoxicity and Anti-inflammatory Efficacy of the Formula in BV2 Microglia Cells

[0262] As a result of combining Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in various ratios, no cytotoxicity was observed when the proportion of Magnolia officinalis was 1 / 10 or less relative to the total (Fig. 3a). As a result of combining Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in various ratios, anti-inflammatory efficacy was confirmed by a concentration-dependent decrease in NO production in all combinations (Fig. 3b). Furthermore, when comparing the anti-inflammatory efficacy of the individual extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis with that of the combinations, no synergistic effect of anti-inflammatory efficacy was observed with the combination of the three natural products (Fig. 3b) (Explanation of figure: *** p<0.001vs CON. ### p<0.001 vs 0 (LPS alone)).

[0263] GROUPO.DMeanStd. DeviationStd. Error of MeanCON1.0628620.9966280.9900670.99941410.043530.0137600.6191990.5914890.588080.6139210.57960.037480.01185 Changchul:Hanryeoncho:Hubak (4.5:4.5:1)300.9099770.8784180.8295580.8676050.86540.031720.014181000.9176750.8584410.8540060.9478040.89020.040870.018283001.0617991.0426651.0213691.0501431.0520.022750.01018Changchul:Hallyeoncho:Houbak (8:1:1)300.9174550.852210.8560220.9412430.86850.064760.028961000.9660220.9188480.9509930.9024270.93050.026770.011973001.1211421.0620921.0702291.0338321.0780.034530.01544Changchul:Hallyeoncho:Hupak (1:8:1)300.9367350.8893780.90251.0201230.93950.051170.022881000.8355690.8661020.8836230.9415730.87990.038810.017363000.9578481.0372040.9737921.1156811.0120.065580.02933Changchul:Hallyeoncho:Hupak (1:1:0)300.8744230.8865190.8615940.9910560.89720.053230.023811000.9447990.8810570.9202770.9840190.89760.086660.038763000.8677880.9871711.0980871.172275 1.0480.12080.05404창출300.9107870.9221780.9488850.8798120.91240.025660.011481000.8477020.8809820.8541380.8674220.85860.015510.0069363000.8589220.9225910.8667690.8479430.87620.02920.01306Hallyeoncho300.925930.9233490.9381470.9379070.92660.012640.0056521000.9781730.9112690.8970890.91650.91310.041980.018773000.9449271.027870.9986511.00649810 .033080.01479Hubak300.8777130.9075170.8270870.9374250.87240.052780.023611000.9540481.0021270.9454440.9654050.95240.038730.017323000.7779750.7066310.6997130.765310.73580.034810.01557.

[0264] GROUPO.DStd. DeviationStd. Error of MeanCON10.0857111.614291.0810.7643095.6857198.428571.9391.371Changchul:Hallyeoncho:Hupak (4.5:4.5:1)3075.7714377.1857110.707110051.0142951.914290.63640.4530015.6571416.842860.83840.5929Changchul:Hallyeoncho:Hupak (8:1:1)3068.1857170.071431.3330.942910029.6142930.542860.65660.464330014.413.885710.36370.2571Changchul:Hallyeocho:Hupak (1:8:1)3059.4285760.91.040.735710034.735.80.77780.5530025.2228625.2228600Changchul:Hallyeocho:Hupak (1:1:0)3056.958.914291.4241.00710042.2571443.242860.6970.49293002425.685711.1920.8429Changchul3064.7285768.614292.7481.94310060.3857162.414291.4341.01430017.9285718.671430.52530.3714Hallyeoncho3 055.1714356.057140.62630.442910033.4857134.457140.68690.485730013.9428613.928570.01010.007145Hubak3052.4714353.80.93940.664310016.0142916.214290.14140.13008.8714299.2857140.29290.2071

[0265] Cell experiments have limitations in that they cannot perfectly simulate the complex environment within the brain; to compensate for this, subsequent research was conducted using animal models to explore the optimal formulation ratio.

[0266]

[0267] 3-2. Evaluation of the Efficacy of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in an Animal Model of Scopolamine-Induced Amnesia

[0268] 3.2.1. Evaluation of the efficacy of single extracts and a combination of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in improving memory in an animal model of scopolamine-induced amnesia.

[0269] The ratios of the mixtures were configured identically to those in the cell experiment. Mix (1) indicated in the graph is a mixture prepared with Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in a ratio of 4.5:4.5:1, Mix (2) in a ratio of 8:1:1, Mix (3) in a ratio of 1:8:1, and Mix (4) in a ratio of 1:1:0. It was confirmed that the test system was well established through the fact that the latency time of donepezil (DNZ), the positive control, was significantly higher than that of the scopolamine-alone group in the Passive Avoidance Test (PAT). As a result of evaluating long-term memory ability through PAT, the latency time in Mixes (1), (2), and (3) was higher than that of the single extracts of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis, indicating superior memory ability. Through this, it was confirmed that the mixtures exhibit a synergistic effect in improving memory ability compared to the single extracts (Fig. 4) (Explanation of the drawing: ** p<0.01 vs CON. ### p<0.001, ## p<0.01 vs - (Scopolamine alone)).

[0270] GROUPAcquisition (sec)MeanStd. DeviationStd. Error of MeanCON6023203026283519353330.911.7423.7132- (SCO)3722411959603642466042.214.6954.6471Mix (1)4144606060604034606051.910.7343.3943Mix(2)2960454060602157605548.714.3924.5511Mix(3)2550263437512960403238.411.7683.7214Mix(4)3446484535434860364143.67.7922.4640Window GROUPRetention (sec)MeanStd. DeviationStd. Error of MeanCON300*941698487161145925014411441.38813.7961- (SCO)1521222168*381824253424.27.3792.4595Mix (1)578124086825644477430010788.46727.9758Mix(2)4827679534677312505013210488.43327.965 0Mix(3)30023112892543833378715111590.01028.4638Mix(4)27202668287191941285358.881.7992 5.8671Changchul668730918929704415414980.943.68213.8134Hanryeoncho80476058348093661283267.829.0099.1734Hupak4127698223472626322840.120.3116.4230DNZ3419891216167422307110613412971.43822.5908

[0271]

[0272] 3.2.2. Comparison of Memory-Improving Efficacy of Three Combinations of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in an Animal Model of Scopolamine-Induced Amnesia

[0273] The ratios of the mixtures were configured in the same way as in the previous experiment. Mix (1) was composed of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in a ratio of 4.5:4.5:1, Mix (2) in a ratio of 8:1:1, and Mix (3) in a ratio of 1:8:1. The concentrations of each mixture were set to 100 and 200 mg / kg for evaluation. It was confirmed that the test system was well established by the fact that the latency time of donepezil (DNZ), the positive control, was significantly higher compared to the scopolamine monotherapy group. As a result of evaluating long-term memory ability using PAT, the latency time of the 200 mg / kg group of Mix (1) among the three mixture ratios was significantly increased compared to the scopolamine monotherapy group, thereby confirming that memory ability improved (Fig. 5) (Explanation of the figure: *** p<0.001 vs CON. ### p<0.001 vs - (Scopolamine alone)).

[0274] GROUP Acquisition (sec)MeanStd. DeviationStd. Error of MeanCON3343242421244531363131.28.2702.6153- (SCO)412660363759514652224312.9874.1069Mix (1)1003060556052522422603845.315.3344.84892004232375540432926495240.59.7213.0741Mix(2)1002745582451602860604345.614.6614.63612006060544732244029476045.313.5574.2871Mix(3)1003329416049193243424038.811.3313.58332004030224252202647503736.611.5973.6673DNZ2138253960563854603442.514.2224.4975GROUP Retention (sec)MeanStd. DeviationStd. Error of MeanCON3003003006030014410430030030024197.35730.7870- (SCO)120*54557579794226462453.321.1547.0514Mix (1)1003728204*5813732671224312772.344.11914.7064200245395491300114136300270108166102.32532.3581Mix(2)10032168321135249300651146398.882.81326.187720013842669290581754716017310452.79616.6956Mix(3)10034413474300*323650252739.214.9904.996620050684538149*384060413846.410.8873.6290DNZ30030075633001253008430070192115.31936.4670

[0275]

[0276] 3.2.3. Evaluation of the memory-improving efficacy of a mixture of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark (4.5:4.5:1) in an animal model of scopolamine-induced amnesia

[0277] When evaluating previous experiments comprehensively, Mix (1) showed the most superior efficacy in improving memory; based on this, the efficacy of Mix (1) in improving memory at different concentrations was evaluated. As a result of evaluating long-term memory ability via PAT, the delay time at Mix (1) 200 and 400 mg / kg significantly increased compared to the scopolamine monotherapy group, confirming that memory ability improved (Fig. 6a). To evaluate short-term memory ability, a Y-maze was performed, and it was confirmed that the experimental system was well established as the change behavior (%) of the positive control DNZ significantly increased compared to the scopolamine monotherapy group. However, there was no significant difference at any concentration of Mix (1) (Fig. 6b). In addition, when measuring the number of times each branch was entered during the Y-maze, there was no significant difference among all groups (Fig. 6c) (Explanation of figure: *** p<0.001 vs CON. # p<0.05, ### p<0.001 vs - (scopolamine alone)).

[0278] GROUPAcquisition (sec)MeanStd. DeviationStd. Error of MeanCON4524275423304042242833.710.7613.4028- (SCO)3131313719343639325134.18.0482.5449Mix (1) 503533453146283131336037.39.9893.1589Mix (1) 1003423423328275528224934.111.1403.5228Mix (1) 2005140372829454360334541.19.9493.1462Mix (1) 4003356303843602829405040.711.4413.6180DNZ39512941304243344645407.1022.2460GROUPRetention (sec)MeanStd. DeviationStd. Error of MeanCON3002721701938830021830017130023174.61623.5956- (SCO)302225101*57363138784139.817.6195.8731Mix (1) 503415744326351304041300*54.739.71813.2393Mix (1) 1002848151389132293958300*57.140.32513.4417Mix (1) 20014623881595015530013120130016692.01329.0969Mix (1) 400233138139941113002931382366617583.80226.5006DNZ12318830012411770343002297215694.74829.9618

[0279] GROUPCONSCOMixture (1)DNZ50100200400Spontaneous Alternation (%)68.29339.28648.00045.00067.85764.00077.19366.66740.00047.05950.00037.93149.18064.70669.23141.66778.37867.85737.73654.90265.62561.76553.70461.90544.00060.00057.89561.90572.22252.83036.36437.50064.28645.71453.33369.69755.00033.33357.89557.14345.71447.82657.89531.81847.50060.00051.56348.83766.66766.66745.23848.78044.44468.18266.66745.94671.05353.06153.84649.20637.25570.00071.05387.50050.00064.58366.66747.05958.69649.254Mean69.146.35252.352.956.260.4Std. Deviation7.787.8413.410.312.48.7910.7Std. Error of Mean2.462.484.243.263.932.783.37

[0280] GROUPCONSCOMixture (1)DNZ50100200400Total arm entries (NO.)43302722305259322236323163532826393055533436562327422144385535583037323542264058372559244227664541384443474644394051156553524026525047364869Mean37.540.233.639.544.745.243.6Std. Deviation9.1913.510.714.41311.513.3Std. Error of Mean2.914.273.374.564.13.654.2

[0281]

[0282] 3-3. Evaluation of the Efficacy of Atractylodes lanceolata, Eclipta prostrata, and Magnolia officinalis in an Animal Model of Amyloid Beta-Induced Alzheimer's Disease

[0283] Experiments were conducted to verify the memory-improving efficacy confirmed in an amyloid beta-induced Alzheimer's disease animal model in an amyloid beta-induced amnesia animal model. It was confirmed that the test system was well established by the fact that the latency time of donepezil (DNZ), the positive control, was significantly higher than that of the amyloid beta monotherapy group in the PAT. As a result of evaluating long-term memory ability through PAT, it was confirmed that memory was impaired by amyloid beta, which led to a decrease in latency time, and that the impaired memory ability was significantly improved at all concentrations of Mix (1) 50, 100, 200, and 400 mg / kg (Fig. 7a). It was confirmed that the test system was well established by the fact that the change behavior (%) of DNZ, the positive control, was significantly increased compared to the amyloid beta monotherapy group in the Y-maze test. As a result of evaluating short-term memory ability using Y-maze, it was confirmed that short-term memory ability was improved in the 400 mg / kg administration group of Mix (1) through a significant increase in change behavior (Fig. 7b) (Explanation of figure: * p<0.05, *** p<0.001 vs SHAM. # p<0.05, ## p<0.01, ### p<0.001 vs - (amyloid beta alone)). Also, Fig. 7c shows that Mix (1) 200 mg / kg and 400 mg / kg is effective in alleviating neurological damage caused by amyloid beta while maintaining exploratory behavior close to normal levels, which supports the neuroprotective and cognitive function-improving effects of Mix (1).

[0284] GROUPAcquisition (sec)MeanStd. DeviationStd. Error of MeanSHAM2917403426273132202127.77.0252.2214- (Aβ)1530263934434533332332.19.1582.8962Mix (1) 504933514024212228274033.510.9673.4681Mix (1) 1001830262723262036231924.85.4731.7308Mix (1) 2004129244335232934225433.410.2333.2359Mix (1) 400402222423142253040563510.8933.4448DNZ3750433429301931313033.48.4492.6717GROUPRetention (sec)MeanStd. DeviationStd. Error of MeanSHAM24296553002091993006329313318996.83030.6203- (Aβ)64232*309025364932724949.721.8697.2896Mix (1) 50300300300129911353058202142169102.08032.2804Mix (1) 1003373493003001056330070259155117.81537.2564Mix (1) 20068300852403003003512863300182115.54336.5378Mix (1) 40030019713330030019210625723718822169.72222.0479DNZ32300249921891367624718830018194.62629.9234

[0285] GROUPSHAMAβMixture (1)DNZ50100200400Spontaneous Alternation (%)59.45965.38558.62170.58873.33380.00069.23172.41444.44462.06975.00064.00068.75050.00075.6760*57.89562.50048.14860.71455.17260.52663.15864.00060.00067.64761.53868.75058.06556.66762.96358.33364.70661.11168.42175.00034.14665.45554.54564.28666.66762.50088.23568.08543.47860.87082.35361.90575.00063.63650.00053.12552.94145.83360.00069.23161.29062.96359.57450.00070.96873.68480.00059.37534.48350.00067.74264.70666.66768.421Mean67.453.357.761.364.666.166.7Std. Deviation9.9913.16.947.9610.96.578.81Std. Error of Mean3.164.372.192.523.442.082.79

[0286] GROUPSHAMAβMixture (1)DNZ50100200400Total arm entries (NO.)39283119322215312931222734183932211829303140402727364118333229141920213084*572416261836492525192314463634192627283329492233211734311033534121Mean36.13431.422.32928.520.1Std. Deviation4.916.7813.45.3310.77.855.47Std. Error of Mean1.552.264.221.693.392.481.73

[0287] The present invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same. Specifically, the invention relates to a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising extracts of Atractylodes rhizome, Eclipta prostrata, and Magnolia bark as active ingredients; and a pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases, a food composition for improving brain cognitive function, a feed composition for improving brain cognitive function, and a method for manufacturing the same, comprising Atractylodin, Wedelolactone, and / or Honokiol as active ingredients.

Claims

A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, comprising atractylodin, wedelolactone, and / or honokiol as active ingredients. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein, in claim 1 or 2, one or more are selected from the group consisting of brain cognitive function-related diseases, neurodegenerative diseases, mental health and mood disorders, cognitive impairment and dementia, developmental disorders, nerve damage and trauma, Creutzfeldt-Jakob disease, chronic traumatic encephalopathy (CTE), and epilepsy. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein the neurodegenerative disease is one or more selected from the group consisting of Alzheimer's disease, Parkinson's disease, Dementia with Lewy Bodies, Huntington's disease, Amyotrophic Lateral Sclerosis (ALS), Multiple Sclerosis (MS), Multiple System Atrophy (MSA), and Progressive Supranuclear Palsy (PSP). A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein, in paragraph 3, the mental health and mood disorders are one or more selected from the group consisting of depression, bipolar disorder, schizophrenia, generalized anxiety disorder (GAD), obsessive-compulsive disorder (OCD), and post-traumatic stress disorder (PTSD). A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein, in paragraph 3, the cognitive impairment and dementia are one or more selected from the group consisting of vascular dementia, frontotemporal dementia, mild cognitive impairment (MCI), Creutzfeldt-Jakob disease (CJD), and senile dementia. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein, in paragraph 3, the developmental disorder is one or more selected from the group consisting of Attention Deficit Hyperactivity Disorder (ADHD), Autism Spectrum Disorder (ASD), Learning Disabilities, and Intellectual Disability. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases, wherein, in paragraph 3, the nerve damage and trauma are one or more selected from the group consisting of traumatic brain injury (TBI), stroke, chronic traumatic encephalopathy (CTE), and hypoxic brain injury. A food composition for improving brain cognitive function comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients. A food composition for improving brain cognitive function comprising attractylodin, wedelolactone and / or honokiol as active ingredients. A pharmaceutical composition for preventing, improving, or treating brain cognitive function-related diseases in companion animals, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients. A pharmaceutical composition for the prevention, improvement, or treatment of brain cognitive function-related diseases in companion animals, wherein, in claim 11, the brain cognitive function-related disease of the companion animal is Cognitive Dysfunction Syndrome (CDS), Canine Dementia (Feline Dementia), Cerebrovascular Disease, or Meningoencephalitis. A feed composition for improving brain cognitive function in companion animals, comprising extracts of Atractylodes rhizome, Erythronium japonicum, and Magnolia bark as active ingredients.

Images