Compositions for improving cognitive function and their use in the prevention and treatment of Alzheimer's disease

Abstract

The present invention provides compositions that improve cognitive function, as well as their use in the prevention and treatment of Alzheimer's disease. [Solution] A composition for improving cognitive function is provided, comprising: docosahexaenoic acid: 8,000 to 24,000 parts by mass, phosphatidylserine: 2,400 to 7,200 parts by mass, vitamin D3: 1 part by mass, vitamin B12: 0.5 to 2 parts by mass, vitamin B6: 50 to 150 parts by mass, vitamin B1: 100 to 300 parts by mass, vitamin B9: 4 to 12 parts by mass, taurine: 2,500 to 7,500 parts by mass, and astaxanthin: 500 to 1,500 parts by mass.

Description

Technical Field

[0001] The present invention relates to the field of biopharmaceuticals, and in particular, to a composition for improving cognitive function and its use in the prevention and treatment of Alzheimer's disease.

Background Art

[0002] Alzheimer's disease (AD) and related dementias have a great impact on global public health and the social economy. According to estimates published by the Alzheimer's Disease International in 2018, the number of dementia patients in the world has reached approximately 50 million, and it is expected to double to about 100 million by 2050. AD is the most common neurodegenerative disease in the elderly, characterized by cognitive decline, deposition of Aβ (amyloid-beta), and neurofibrillary changes (tangles of neurofibrils). Other pathological features include progressive degeneration / loss of neurons and synapses, neuroinflammation, and oxidative damage.

[0003] AD is a complex clinical syndrome, characterized by progressive decline across multiple cognitive domains such as memory, language, executive function, visuospatial ability, personality, and behavior. Such cognitive decline ultimately renders patients unable to perform instrumental and / or basic activities of daily living independently.

[0004] AD is a progressive deterioration process. Pathological changes may begin before the clinical symptoms become apparent, and then, after passing through the mild cognitive impairment (MCI) stage, it finally progresses to significant dementia symptoms.

[0005] Currently, drug treatment options for Alzheimer's disease (AD) are limited and mainly classified into two types: (1) Cholinesterase inhibitors (such as donepezil, rivastigmine, and galantamine): Applicable to patients with mild, moderate, or severe AD and Parkinson's disease-associated dementia. (2) N-methyl-D-aspartate (NMDA) receptor antagonists (such as memantine): As non-competitive NMDA receptor antagonists and dopamine agonists, they are mainly used in patients with moderate to severe AD, especially those with attention and arousal problems. However, these drugs have limited efficacy and can cause side effects, so the emergence of new drugs for the prevention and treatment of AD is anticipated. [Overview of the project] [Problems that the invention aims to solve]

[0006] The object of the present invention is to provide a composition that improves cognitive function and its use in the prevention and treatment of Alzheimer's disease. [Means for solving the problem]

[0007] As a first aspect, the present invention provides compositions for improving cognitive function.

[0008] The compositions provided by the present invention further have functions such as treating and / or preventing Alzheimer's disease, a neurodegenerative disease, improving the symptoms of neurodegenerative diseases, and reducing the toxicity to nerve cells caused by β-amyloid protein. Here, the neurodegenerative disease is a disease that is particularly associated with cognitive decline, such as Alzheimer's disease.

[0009] The composition is composition A, composition B, or composition C, wherein composition A consists of a neuronal cell membrane protectant and a nerve repair factor, composition B consists of the neuronal cell membrane protectant, the nerve repair factor and a protective enhancer, and composition C consists of the nerve repair factor.

[0010] The aforementioned neuronal cell membrane protective agent comprises all or part of the following components: omega-3 fatty acids and phospholipid compounds.

[0011] The omega-3 fatty acid is docosahexaenoic acid (DHA) and / or eicosapentaenoic acid (EPA), and the phospholipid compound is phosphatidylserine, phosphatidylcholine, and / or phosphatidylethanolamine, and the composition significantly improves the stability and integrity of nerve cell membranes.

[0012] The aforementioned nerve repair factor includes all or part of the following components: vitamin D group and vitamin B group.

[0013] The vitamin D group is vitamin D2 and / or vitamin D3, and the vitamin B group is selected from vitamin B12, vitamin B6, vitamin B1 and / or vitamin B9, and the composition promotes the growth and repair of nerve cells.

[0014] The aforementioned protective enhancer comprises all or part of the following components: taurine and carotenoids.

[0015] Here, the taurine may be selectively partially or completely substituted with a taurine derivative (e.g., N-acetyltaurine), and the carotenoid is astaxanthin, β-carotene, and / or lutein, and the composition improves the antioxidant capacity and energy metabolism of nerve cells.

[0016] Furthermore, the neuronal cell membrane protective agent may be any of the following:

[0017] (a1) In accordance with animal experiments, the following parts by mass of omega-3 fatty acids and the phospholipid compound are used: The aforementioned omega-3 fatty acid (e.g., DHA): 1 to 10 parts by mass (e.g., 3.3 parts by mass) The phospholipid compound (e.g., phosphatidylserine): 1 part by mass (a2) In accordance with cell experiments, the following molar portions of omega-3 fatty acids and the phospholipid compounds are composed: The aforementioned omega-3 fatty acid (e.g., DHA): 0.5 to 2 moles (e.g., 1 mole) The phospholipid compound (e.g., phosphatidylserine): 1 mole part In one embodiment of the present invention, the neuronal cell membrane protective agent is composed of DHA and phosphatidylserine in a mass ratio of 3.3:1 (corresponding to animal experiments).

[0018] In another embodiment of the present invention, the neuronal cell membrane protective agent is composed of DHA and phosphatidylserine in a molar ratio of 1:1 (corresponding to cell experiments).

[0019] Furthermore, the nerve repair factor may be any of the following:

[0020] (b1) Composed of the following parts by mass of vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9: Vitamin D (e.g., vitamin D3): 0.1 parts by mass Vitamin B12: 0.05 to 1 part by mass (e.g., 0.1 part by mass) Vitamin B6: 5-50 parts by mass (e.g., 10 parts by mass) Vitamin B1: 10-100 parts by mass (for example, 20 parts by mass) Vitamin B9: 0.4-4 parts by mass (e.g., 0.8 parts by mass) (b2) Composed of the following molar portions of vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9: Vitamin D (e.g., vitamin D3): 1 mole Vitamin B12: 0.5-2 moles (e.g., 1 mole) Vitamin B6: 0.5-2 moles (e.g., 1 mole) Vitamin B1: 0.5-2 moles (e.g., 1 mole) Vitamin B9: 0.5-2 moles (e.g., 1 mole) In one embodiment of the present invention, the nerve repair factor is composed of vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 with a mass ratio of 0.1:0.1:10:20:0.8 (corresponding to animal experiments).

[0021] In another embodiment of the present invention, the nerve repair factor is composed of vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 with a molar ratio of 1:1:1:1:1 (corresponding to cell experiments).

[0022] Furthermore, the protection enhancer can be any of the following.

[0023] (c1) It is composed of the following parts by mass of taurine and the carotenoid: Taurine: 1 to 10 parts by mass (for example, 5 parts by mass) The carotenoid (for example, astaxanthin): 1 part by mass (c2) It is composed of the following parts by mole of taurine and the carotenoid: Taurine: 0.5 to 2 parts by mole (for example, 1 part by mole) The carotenoid (for example, astaxanthin): 1 part by mole Here, the taurine can be selectively partially or completely replaced by a taurine derivative (for example, N-acetyltaurine).

[0024] In one embodiment of the present invention, the protection enhancer is composed of taurine and astaxanthin with a mass ratio of 5:1 (corresponding to animal experiments).

[0025] In another embodiment of the present invention, the protection enhancer is composed of taurine and astaxanthin with a molar ratio of 1:1 (corresponding to cell experiments).

[0026] Furthermore, the composition is any of the following.

[0027] (d1) consists of the following parts by mass of omega-3 fatty acids (e.g., DHA), the phospholipid compound (e.g., phosphatidylserine), vitamin D, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and the carotenoid (e.g., astaxanthin): Omega-3 fatty acids (e.g., DHA): 8,000 to 24,000 parts by mass (e.g., 16,000 parts by mass) The phospholipid compound (e.g., phosphatidylserine): 2400 to 7200 parts by mass (e.g., 4800 parts by mass) Vitamin D (e.g., vitamin D3): 1 part by mass Vitamin B12: 0.5 to 2 parts by mass (e.g., 1 part by mass) Vitamin B6: 50-150 parts by mass (for example, 100 parts by mass) Vitamin B1: 100-300 parts by mass (for example, 200 parts by mass) Vitamin B9: 4-12 parts by mass (for example, 8 parts by mass) Taurine: 2500-7500 parts by mass (e.g., 5000 parts by mass) The carotenoid (e.g., astaxanthin): 500 to 1500 parts by mass (e.g., 1000 parts by mass) (d2) consists of the following parts by mass: omega-3 fatty acids (e.g., DHA), the phospholipid compound (e.g., phosphatidylserine), vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9: Omega-3 fatty acids (e.g., DHA): 8,000 to 24,000 parts by mass (e.g., 16,000 parts by mass) The phospholipid compound (e.g., phosphatidylserine): 2400 to 7200 parts by mass (e.g., 4800 parts by mass) Vitamin D (e.g., vitamin D3): 1 part by mass Vitamin B12: 0.5 to 2 parts by mass (e.g., 1 part by mass) Vitamin B6: 50-150 parts by mass (for example, 100 parts by mass) Vitamin B1: 100-300 parts by mass (for example, 200 parts by mass) Vitamin B9: 4-12 parts by mass (for example, 8 parts by mass) (d3) Composed of the following parts by mass of vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9: Vitamin D (e.g., vitamin D3): 1 part by mass Vitamin B12: 0.5 to 2 parts by mass (e.g., 1 part by mass) Vitamin B6: 50-150 parts by mass (for example, 100 parts by mass) Vitamin B1: 100-300 parts by mass (for example, 200 parts by mass) Vitamin B9: 4-12 parts by mass (for example, 8 parts by mass) (d4) consists of the following molar portions of the omega-3 fatty acids, phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and carotenoids: The aforementioned omega-3 fatty acid (e.g., DHA): 0.5 to 2 moles (e.g., 1 mole) The phospholipid compound (e.g., phosphatidylserine): 0.5 to 2 moles (e.g., 1 mole) Vitamin D (e.g., vitamin D3): 1 mole Vitamin B12: 0.5-2 moles (e.g., 1 mole) Vitamin B6: 0.5-2 moles (e.g., 1 mole) Vitamin B1: 0.5-2 moles (e.g., 1 mole) Vitamin B9: 0.5-2 moles (e.g., 1 mole) Taurine: 0.5-2 moles (e.g., 1 mole) The carotenoid (e.g., astaxanthin): 0.5 to 2 moles (e.g., 1 mole) (d5) consists of the omega-3 fatty acids, the phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in molar portions less than or equal to: The aforementioned omega-3 fatty acid (e.g., DHA): 0.5 to 2 moles (e.g., 1 mole) The phospholipid compound (e.g., phosphatidylserine): 0.5 to 2 moles (e.g., 1 mole) Vitamin D (e.g., vitamin D3): 1 mole Vitamin B12: 0.5-2 moles (e.g., 1 mole) Vitamin B6: 0.5-2 moles (e.g., 1 mole) Vitamin B1: 0.5-2 moles (e.g., 1 mole) Vitamin B9: 0.5-2 moles (e.g., 1 mole) (d6) Composed of the following molar portions of vitamin D, vitamin B12, vitamin B6, vitamin B1, and vitamin B9: Vitamin D (e.g., vitamin D3): 1 mole Vitamin B12: 0.5-2 moles (e.g., 1 mole) Vitamin B6: 0.5-2 moles (e.g., 1 mole) Vitamin B1: 0.5-2 moles (e.g., 1 mole) Vitamin B9: 0.5-2 moles (e.g., 1 mole) In (d1) and (d4) above, the taurine may be selectively partially or completely substituted with a taurine derivative (e.g., N-acetyltaurine).

[0028] In one embodiment of the present invention, the composition comprises DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and astaxanthin in a mass ratio of 16000:4800:1:1:100:200:8:5000:1000 (corresponding to the combination of Vit+DP+TA in animal experiments, i.e., nerve repair factors, neuronal cell membrane protectants, and protective enhancers).

[0029] In another embodiment of the present invention, the composition comprises DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in a mass ratio of 16000:4800:1:1:100:200:8 (corresponding to the animal experiment Vit+DP, i.e., a combination of nerve repair factors and neuronal cell membrane protectants).

[0030] In yet another embodiment of the present invention, the composition comprises vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in a mass ratio of 1:1:100:200:8 (corresponding to animal experiment Vit, i.e., nerve repair factors).

[0031] In yet another embodiment of the present invention, the composition consists of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and astaxanthin in a molar ratio of 1:1:1:1:1:1:1:1:1 (corresponding to cell experiment Vit+DP+TA).

[0032] In yet another embodiment of the present invention, the composition consists of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in a molar ratio of 1:1:1:1:1:1:1 (corresponding to the cell experiment Vit+DP).

[0033] In yet another embodiment of the present invention, the composition consists of vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in a molar ratio of 1:1:1:1:1 (corresponding to the cell experiment Vit).

[0034] Secondly, the present invention relates to the use of the composition described in the first aspect in the manufacture of a product for improving cognitive function or for improving cognitive function.

[0035] Here, the improvement in cognitive function may include spatial learning and memory abilities, novel object recognition abilities, and so on.

[0036] In a third aspect, the present invention relates to any of the following uses of the compositions described in the first aspect.

[0037] (A1) Manufacturing of products for the treatment and / or prevention of neurodegenerative diseases, or for the treatment and / or prevention of neurodegenerative diseases.

[0038] (A2) Manufacturing of products for improving symptoms of neurodegenerative diseases, or improving symptoms of neurodegenerative diseases.

[0039] Here, the neurodegenerative disease may be a cognitive impairment disease, such as Alzheimer's disease.

[0040] Fourthly, the present invention relates to the use of the composition described in the first aspect in the manufacture of a product that can reduce the toxicity to nerve cells caused by neurotoxic proteins, or to the reduction of the toxicity to nerve cells caused by neurotoxic proteins.

[0041] Here, the neurotoxic protein includes, but is not limited to, β-amyloid protein, tau protein, TDP-43 protein, and α-synuclein protein. In one embodiment of the present invention, the neurotoxic protein is β-amyloid protein.

[0042] Here, the nerve cells include, but are not limited to, primary nerve cells, SH-SY5Y cells, and iPSC pluripotent stem cell-induced nerve cells. In one embodiment of the present invention, the nerve cells are SH-SY5Y cells.

[0043] Fifthly, the present invention relates to a product having any of the following functions: (B1) Improvement of cognitive function (B2) Treatment and / or prevention of neurodegenerative diseases (B3) Improvement of symptoms of neurodegenerative disease (B4) Reduction of the toxic effects on nerve cells caused by neurotoxic proteins In (B2) and (B3), the neurodegenerative disease may be a cognitive impairment disease, such as Alzheimer's disease.

[0044] In (B4), the neurotoxic proteins include, but are not limited to, β-amyloid protein, tau protein, TDP-43 protein, and α-synuclein protein. The nerve cells include, but are not limited to, primary nerve cells, SH-SY5Y cells, and iPSC pluripotent stem cell-induced nerve cells.

[0045] The product according to the present invention includes the composition described in the first aspect.

[0046] In the aforementioned related aspects, the product may be a nutritional fortifier or a pharmaceutical. As a nutritional fortifier, it may be used alone in beverages, injections, or food ingredients, or in combination with other nutritional supplements or therapeutic agents. It may also be formulated and used with pharmaceutical compounds, carriers, or excipients with excellent delivery capabilities (i.e., suitable for delivery to a subject). Such compositions typically include the fruit-based nutritional supplement and pharmaceutical carrier of the present invention. In this specification, “pharmaceutical carrier” tends to further include pharmaceutically suitable solvents, dispersion media, coatings, antimicrobial and antifungal compounds, isotonic and sustained-release compounds, and similar items. Preferred examples of such carriers or diluents include, but are not limited to, water, physiological saline, Ringer's solution, glucose solution, and 5% human serum albumin. Liposomes and non-volatile oily carriers may also be used. The preparation of pharmaceutically active substances with these media and compounds is known in the art.

[0047] Furthermore, as a sixth aspect, the present invention relates to a method for improving cognitive function and includes the following steps: administering an effective amount of the composition described in the first aspect to a subject.

[0048] Furthermore, as a seventh aspect, the present invention relates to a method for treating and / or preventing neurodegenerative diseases, comprising the following steps: administering an effective amount of the composition described in the first aspect to a subject.

[0049] Furthermore, as an eighth aspect, the present invention relates to a method for improving symptoms of neurodegenerative diseases, comprising the following steps: administering an effective amount of the composition described in the first aspect to a subject.

[0050] Furthermore, as a ninth aspect, the present invention relates to a method for reducing the toxicity to nerve cells caused by neurotoxic proteins, and comprises the following steps: treating nerve cells damaged or to be damaged by neurotoxic proteins with an effective amount of the composition described in the first aspect. [Effects of the Invention]

[0051] In relation to the above-mentioned aspects, the subject is a patient with a neurodegenerative disease. The neurodegenerative disease may be a cognitive impairment disease, such as Alzheimer's disease. The neurotoxic proteins include, but are not limited to, β-amyloid protein, tau protein, and α-synuclein protein. The nerve cells include, but are not limited to, primary nerve cells, SH-SY5Y cells, and iPSC pluripotent stem cell-induced nerve cells.

[0052] In each of the above-mentioned related aspects, the improvement of Alzheimer's disease symptoms can manifest specifically as behavioral improvements and / or pathological improvements. Of these, the behavioral improvements may include improvements in cognitive function. The pathological improvements may include: a decrease in Aβ deposition and microglial cell activation in the hippocampal dentate gyrus (DG) region, a decrease in APP and phosphorylated APP deposition in the DG region, a decrease in APP and tau protein levels, and a restoration of hippocampal dentate gyrus lilin protein secretion.

[0053] This invention has demonstrated, through in vitro experiments, that the compositions provided by this invention can significantly improve the pathological state of an Alzheimer's disease model. In vitro experiments showed that the compositions of this invention can reduce the toxic effects of Aβ in SH-SY5Y cells. In vitro experiments showed that the compositions of this invention can significantly improve the cognitive abilities of 5xFAD mice, based on cognitive ability and pathological sections. Overall, the compositions of this invention have a remarkable therapeutic effect on improving cognitive function and simultaneously have a certain preventive and therapeutic effect on Alzheimer's disease.

[0054] Furthermore, the components of the composition provided by this invention are nutritional fortifiers / supplements, with a milder mechanism of action, fewer side effects, and suitability for long-term use. Therefore, it is beneficial for overall health maintenance and may contribute to improving not only AD symptoms but also age-related decline in systemic organ function. It also reduces the risks of polypharmacy, making it particularly suitable for elderly patients who need to take many medications. [Brief explanation of the drawing]

[0055] [Figure 1] This study demonstrates that a combination of nerve repair factors and neuronal cell membrane protectants (Vit+DP) can mitigate the toxic effects of Aβ in SH-SY5Y cells. [Figure 2] This study demonstrates that a combination of the neuroprotective agents (TAs) taurine and astaxanthin can mitigate the toxic effects of Aβ in SH-SY5Y cells. [Figure 3] This study demonstrates that a combination of nerve repair factors, neuronal cell membrane protectants, and enhancing protective agents (Vit+DP+TA) can mitigate the toxic effects of Aβ in SH-SY5Y cells. [Figure 4] This study evaluated the effects of Vit+DP+TA treatment on spatial memory and cognitive function using the Y-maze test. A represents a typical heatmap image of the Y-maze test, B represents the number of new arm entries, and C represents the spontaneous alternation rate. [Figure 5] This study evaluated the impact of Vit+DP+TA treatment on cognitive function using a novel object recognition test. A is a representative heatmap image of the novel object recognition (NOR) test, where red circles represent new objects and black circles represent known objects. B shows the percentage of time spent searching for new objects, with data presented as mean ± standard error (SEM). [Figure 6] The Morris Water Maze Test was used to evaluate the effects of Vit+DP+TA treatment on cognitive function. A represents the typical swimming trajectory of the mouse during the learning or memory phase, B represents the escape latency and swimming path during the learning phase, C represents the escape latency during the memory phase, and D represents the number of times the platform was crossed during the memory phase. [Figure 7] This study shows that treatment with nerve repair factors (Vit) reduces Aβ deposition and microglial cell activation in the hippocampal dentate gyrus. [Figure 8] This shows how nerve repair factor (Vit) therapy reduces APP and phosphorylated APP deposition in the hippocampal dentate gyrus. [Figure 9]This study demonstrates that nerve repair factor (Vit) therapy can reduce levels of pathologically phosphorylated APP and Tau proteins in 5xFAD mice. [Figure 10] This study shows that treatment with nerve repair factors (vitamins) increases the expression of neuroprotective lilin protein (Reelin). [Modes for carrying out the invention]

[0056] The present invention will be described in more detail below by combining specific embodiments. The embodiments presented are for illustrative purposes only and do not limit the scope of the present invention. The embodiments provided below will serve as a guide for a person ordinary in the art to make further improvements and do not limit the present invention in any sense.

[0057] Unless otherwise specified, the experimental methods in the following examples were carried out according to conventional methods, in accordance with the techniques or conditions described in the relevant art literature, or in accordance with the product instructions. Unless otherwise specified, all materials and reagents used in the following examples can be commercially available.

[0058] The components of the composition provided by the present invention include all or some of the following: DHA, taurine, phosphatidylserine, astaxanthin, vitamin B12, vitamin D3, vitamin B1, vitamin B6, and vitamin B9. DHA and phosphatidylserine are neuronal cell membrane protectants (abbreviated as DP), vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 are nerve repair factors (abbreviated as Vit), and taurine and astaxanthin are enhancing protective agents (abbreviated as TA).

[0059] The specific classifications and functions of these compositions are shown in Table 1.

[0060] [Table 1] The following examples verify the effectiveness of the compositions provided by the present invention in improving cognitive function and preventing / treating Alzheimer's disease (AD) from two perspectives: in vitro experiments (cell experiments) and in vivo experiments (mouse experiments).

[0061] Example 1: In vitro efficacy evaluation of the composition of the present invention The cell line used in the in vitro cell experiment of this invention is the SH-SY5Y cell line. As a first-choice cell model for AD research, SH-SY5Y cells possess neuronal characteristics, are capable of differentiation, and provide an ideal platform for AD research. Their immortalization properties ensure experimental stability and reproducibility. SH-SY5Y cells are highly sensitive to Aβ toxicity and are a powerful tool for studying important pathological mechanisms of AD. As human-derived cells, they are closer to human neurons than other animal cells and may be able to more accurately reflect some pathological processes of AD. The SH-SY5Y cell line used in this example is manufactured by BeNa Culture Collection (Shanghai, China).

[0062] 1. Combination of nerve repair factors and nerve cell membrane protectants (Vit+DP) SH-SY5Y cells were seeded in a 96-well plate containing complete medium and cultured at 37°C for 24 hours. Then, they were divided into the following groups and each group was treated for 24 hours.

[0063] Blank control group: SH-SY5Y cells only Positive control group (Aβ-treated): SH-SY5Y cells + 1 μM Aβ-42 (Sigma-Aldrich # A9810, same applies below) Nerve repair factor group (Aβ + Vitamin treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM nerve repair factor Neuronal cell membrane protective agent group (Aβ+DP treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM neuronal cell membrane protective agent Nerve repair factor + neuronal membrane protective agent group (Aβ + Vit + DP treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM nerve repair factor + 10 μM neuronal membrane protective agent Note: In each group, "1 μM Aβ-42" means that the final concentration of Aβ-42 in the SH-SY5Y cell culture system is 1 μM, "10 μM nerve repair factor" means that the final concentrations of vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 in the SH-SY5Y cell culture system are all 10 μM, and "10 μM neuronal membrane protectant" means that the final concentrations of DHA and phosphatidylserine in the SH-SY5Y cell culture system are both 10 μM.

[0064] After treatment, SH-SY5Y cells from each group were incubated with MTT (final concentration in the incubation system: 0.5 mg / mL) for 4 hours, and then incubated for another 15 minutes at 37°C with 10% SDS solution. Absorbance at 560 nm was measured using an enzyme labeling device (Spectra Max M2, Molecular Devices, San Jose, CA, USA). Specifically, the MTT Cell Proliferation Assay Kit (VOBOSCIENCE) was used to complete the process.

[0065] Calculating relative cell vitality: Cell viability (%) = (OD560 value of treated group - OD560 value of blank control) / (OD560 value of control group - OD560 value of blank control) × 100% Here, "treatment group OD560 value" is the 560nm absorbance of experimentally treated cell samples, "blank control OD560 value" is the 560nm absorbance of wells containing only culture medium and reagents but no cells, and "control group OD560 value" is the 560nm absorbance of untreated normal cells.

[0066] Each group was set up five times, and the results are shown as the mean ± standard deviation.

[0067] The results are shown in Figure 1. As can be seen from the figure, there were differences in cell viability under different treatment conditions, with the blank control group (SH-SY5Y cells only) having the highest cell viability at almost 100%. The cell viability of the positive control group (Aβ treatment) decreased significantly to about 60%, showing a very significant difference compared to the blank control group (****p<0.0001). Cell viability increased slightly in both the nerve repair factor group (Aβ+Vit treatment) and the neuronal membrane protective agent group (Aβ+DP treatment), but there was no significant difference compared to the Aβ treatment group. The nerve repair factor + neuronal membrane protective agent group (Aβ+Vit+DP treatment) showed the best therapeutic effect, with a clearly higher cell viability of about 80% compared to the Aβ treatment group, and was the only treatment group to show a statistically significant difference compared to the Aβ treatment group (****p<0.0001). These results indicate that the combined use of nerve repair factors (Vit) and neuronal membrane protectants (DP) has a synergistic effect in protecting cells from Aβ toxicity.

[0068] 2. Combinations of enhancing protective agents (TA) SH-SY5Y cells were seeded in a 96-well plate containing complete medium and cultured at 37°C for 24 hours. Then, they were divided into the following groups and each group was treated for 24 hours.

[0069] Blank control group: SH-SY5Y cells only Positive control group (Aβ-treated): SH-SY5Y cells + 1 μM Aβ-42 Taurine group (Aβ + Taur treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM taurine Astaxanthin (Asta) group (Aβ + Asta treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM astaxanthin Taurine + Astaxanthin group (Aβ + Taur + Asta treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM taurine + 10 μM astaxanthin Note: The "1 μM Aβ-42" in each group is the final concentration in the SH-SY5Y cell culture system, and the "10 μM taurine" and "10 μM astaxanthin" are their respective final concentrations.

[0070] After treatment, SH-SY5Y cells from each group were incubated with MTT (final concentration 0.5 mg / mL) for 4 hours, then incubated with 10% SDS solution at 37°C for a further 15 minutes. Cell vitality was calculated by measuring the absorbance at 560 nm using an enzyme labeling device (Spectra Max M2, Molecular Devices, San Jose, CA, USA). Details are as described above. Each group was set up five times, and the results are shown as mean ± standard deviation.

[0071] The results are shown in Figure 2. As can be seen from the figure, there were differences in cell viability under different treatment conditions. The blank control group (SH-SY5Y cells only) had the highest cell viability, at almost 100%. The positive control group (Aβ treatment) had the lowest cell viability, at about 60%, indicating that Aβ has a clear toxic effect on cells (****p<0.0001). The taurine group (Aβ+Taur treatment) showed a slight increase in cell viability, but the effect was not clear. The astaxanthin group (Aβ+Asta treatment) showed a higher cell viability than Aβ alone, reaching about 70%. The taurine + astaxanthin group (Aβ+Taur+Asta treatment) showed an even higher cell viability, reaching about 75-80%, which was a very significant difference compared to the Aβ-treated group (****p<0.0001). This indicates that the combination of the two substances is superior to each other.

[0072] Astaxanthin or taurine alone have been shown to some extent to suppress Aβ toxicity, and combinations of the two are more effective than using either one alone.

[0073] 3. Combinations of nerve repair factors, nerve cell membrane protectants, and enhancing protectants (Vit + DP + TA) SH-SY5Y cells were seeded in a 96-well plate containing complete medium and cultured at 37°C for 24 hours. Then, they were divided into the following groups and each group was treated for 24 hours.

[0074] Blank control group: SH-SY5Y cells only Positive control group (Aβ-treated): SH-SY5Y cells + 1 μM Aβ-42 Nerve repair factor + neuronal membrane protective agent group (Aβ + Vit + DP treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM nerve repair factor + 10 μM neuronal membrane protective agent Enhancement and protective agent group (Aβ+TA treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM taurine + 10 μM astaxanthin Nerve repair factors + neuronal membrane protectants + enhancing protective agents (Aβ + Vit + DP + TA treatment): SH-SY5Y cells + 1 μM Aβ-42 + 10 μM nerve repair factors + 10 μM neuronal membrane protectants + 10 μM taurine + 10 μM astaxanthin Note: For each group, "1 μM Aβ-42" is the final concentration of the SH-SY5Y cell culture system, "10 μM nerve repair factor" is the final concentration of vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9, each at 10 μM, "10 μM neuronal membrane protectant" is the final concentration of DHA and phosphatidylserine, each at 10 μM, and "10 μM taurine" and "10 μM astaxanthin" are their respective final concentrations.

[0075] After treatment, SH-SY5Y cells from each group were incubated with MTT (final concentration 0.5 mg / mL) for 4 hours, then incubated with 10% SDS solution at 37°C for a further 15 minutes. Cell vitality was calculated by measuring the absorbance at 560 nm using an enzyme labeling device (Spectra Max M2, Molecular Devices, San Jose, CA, USA). Details are as described above. Each group was set up five times, and the results are shown as mean ± standard deviation.

[0076] The results are shown in Figure 3. As can be seen from the figure, there were differences in cell viability under different treatment conditions, with the blank control group (SH-SY5Y cells only) having the highest cell viability at almost 100%. The cell viability of the positive control group (Aβ treatment) decreased significantly to about 60%, showing an extremely significant difference compared to the blank control group (****p<0.0001). Cell viability increased in both the nerve repair factor + nerve cell membrane protectant group (Aβ+Vit+DP treatment) and the augmentative protective agent group (Aβ+TA treatment). The nerve repair factor + nerve cell membrane protectant + augmentative protective agent group (Aβ+Vit+DP+TA treatment) showed the best therapeutic effect, with a cell viability of approximately 90% in this group, which was clearly higher than the Aβ monotherapy group, and the difference from the blank control group was minimal. This group showed an extremely significant difference compared to the Aβ monotherapy group (****p<0.0001).

[0077] Of particular note is that the average cell viability was 62.6% in the Aβ monotherapy group, 71.0% in the Aβ+Vit+DP group, 77.8% in the Aβ+TA group, and 89.2% in the Aβ+Vit+DP+TA group. Calculations show that compared to the Aβ monotherapy group, the Aβ+Vit+DP group showed an 8.4% improvement in survival rate, the Aβ+TA group a 15.2% improvement, and the Aβ+Vit+DP+TA group a 26.6% improvement. This exceeds the combined improvement in the Aβ+Vit+DP and Aβ+TA groups, demonstrating a significant synergistic effect from the combination of TA and Vit+DP, and providing guidance for a new strategy in the treatment of neurodegenerative diseases.

[0078] Example 2 Evaluation of the efficacy of the composition of the present invention in the body The in vivo mouse experiments of this invention used a 5xFAD disease model mouse (Cat# 034848-JAX, RRID: MMRRC_034848-JAX). This model mouse has five familial Alzheimer's disease (AD) mutations. The 5xFAD mouse has human APP(695) K670N / M67L (Sweden), I716V (Florida), and V717I (London) mutations in its brain, as well as human PSEN1 gene M146L and L286V mutations. The 5xFAD mouse is a transgenic model in which the pathological features of Alzheimer's disease progress rapidly. These mice develop amyloid plaques at 2 months of age and show significant neuronal loss at 4-5 months of age. Cognitive and behavioral abnormalities usually appear gradually from 2-3 months of age, become apparent at 4-5 months of age, and continue to progress thereafter. The 5xFAD model is widely used to study the pathological mechanisms of Alzheimer's disease, test potential treatment strategies, and investigate neuroinflammatory and neurodegenerative processes.

[0079] 1. Behavioral experiments In this animal experiment, a total of 14 5xFAD mice and 7 wild-type littermate mice (C57BL / 6 mice, all female) were used. All mice were 5 months old. The 5xFAD mice were randomly divided into a positive treatment group (N=7), a positive control group (N=7), and a wild-type mouse group (N=7). The positive treatment group received daily oral administration of a nerve repair factor + nerve cell membrane protectant + enhancing protective agent composition (Vit+DP+TA) (see Table 1 for the dosage of each component). The positive control group received oral administration of a pure solvent (1% Tween 80 + 1% methylcellulose + 98% water, by mass ratio) for a period of 3 months. Subsequently, animal behavioral experiments were conducted.

[0080] As part of animal behavioral experiments, we investigated changes in cognitive function using a Y-maze with a novel arm, a Y-maze with spontaneous alternation, a novel object recognition, and a Morris water maze.

[0081] 1) Y-maze - New arm and spontaneous rotation The Y-maze experiment evaluates the short-term spatial cognition and memory abilities of mice. The Y-maze apparatus consists of three arms of equal length (30 cm long, 8 cm wide, and 15 cm high) spaced 120 degrees apart.

[0082] Before the novel arm exploration experiment in the Y-maze, the illumination of the experimental area was adjusted to approximately 100 Lux, and the mice were moved to the experimental area and allowed to acclimate for at least 30 minutes. One arm of the Y-maze was blocked off with a partition to create the novel arm, and then the mouse was placed with its back to the starting arm of the Y-maze and allowed to explore freely for 5 minutes. After the training was complete, the mouse was returned to its cage. After a 15-minute interval, the partition was removed to open the novel arm, and the mouse was again placed with its back to the starting arm and allowed to explore freely for 5 minutes. An entry into an arm was determined when the mouse's limbs were completely inside any of the arms, and the number of times the mouse entered the novel arm and the total number of times it entered all arms were manually recorded. After the experiment, the open field was cleaned with 75% ethanol to remove any lingering odor from the mice.

[0083] Prior to the spontaneous rotation experiment in the Y-maze experiment, the illumination of the experimental area was adjusted to approximately 100 Lux, and the mice were moved to the experimental area and allowed to acclimate for at least 30 minutes. During the experiment, the mice were placed with their backs to one of the arms and allowed to explore freely for 8 minutes. Entry was determined when all four limbs of the mouse were completely inside one of the arms, and the order of entry was recorded. After the experiment, the open field was cleaned with 75% ethanol to remove any lingering odor from the mice.

[0084] The results are shown in Figure 4, which consists of three parts: a heatmap, the number of entries into new arms, and the spontaneous alternation rate. From the heatmap, it was seen that the wild-type mouse group was equally active in all three arms, the positive control group (pure solvent) showed decreased activity in new arms, and the positive treatment group (Vit+DP+TA treatment) recovered activity in new arms to a level close to that of the wild-type mouse group. In both the number of entries into new arms and the spontaneous alternation rate, the positive control group (pure solvent) was significantly lower than the wild-type mouse group, indicating a decline in cognitive function, but the Vit+DP+TA treatment group showed clear improvement. These results clearly demonstrate that Vit+DP+TA treatment significantly improves spatial memory and working memory.

[0085] 2) New object recognition The new object recognition experiment was conducted in a 40cm x 40cm x 40cm open field box to evaluate the exploration and cognitive abilities of mice. The experiment was conducted over three days. On the first day, the mice were allowed to move freely for 5 minutes as an adaptation period. On the second day, they were allowed to explore two identical objects for 5 minutes as a training period. On the third day, as a test period, they were first allowed to explore two identical objects for 5 minutes, then, after a 15-minute interval, one of the objects was replaced with a new object, and they were allowed to explore for another 5 minutes. The exploration time was analyzed using Tracking Master V3.1.62 software, and the cognitive index was calculated as: New object exploration time / (New object exploration time + Old object exploration time) × 100%. The illumination level of the experimental environment was always maintained at approximately 100 Lux, and the open field was cleaned with 75% ethanol after each experiment.

[0086] The results are shown in Figure 5, which includes a heatmap and cognitive index as results of the Novel Object Recognition (NOR) test. The heatmap shows that the wild-type mouse group and the positive treatment group (Vit+DP+TA treatment) were more active around the novel object, while the positive control group (pure solvent) showed less interest in the novel object. The cognitive index graph also shows that the recognition ability of the positive control group (pure solvent) was significantly lower than that of the wild-type mouse group, and the mice treated with Vit+DP+TA showed a considerable degree of recovery in their recognition ability.

[0087] These results clearly demonstrate that Vit+DP+TA treatment significantly improves object recognition ability and cognitive function in mice.

[0088] 3.Morris water maze The Morris water maze experiment evaluates the learning, memory, and spatial cognitive abilities of mice. It uses a circular tank 1.2m in diameter and 0.5m high, divided into four quadrants, one of which contains a hidden platform 5cm in diameter. The experiment is divided into two phases: a learning phase (platform concealment phase) and a memory phase (no platform phase). The learning phase is conducted four times daily, each time for 60 seconds, recording the time and path taken by the mouse to find the hidden platform. The memory phase is conducted 24 hours after the end of the learning phase, with the platform removed. Mice are allowed to swim freely for 60 seconds, and their swimming path, the number of times they passed the original platform location, and the time spent in the original platform quadrant are recorded. The illumination of the experimental environment is always maintained at approximately 100 Lux, and food-grade titanium dioxide powder is added to the water in the tank to make it milky white. Mice are allowed to acclimate to the experimental environment for at least 30 minutes. These data are used to analyze the mice's spatial learning and memory abilities.

[0089] The results are shown in Figure 6. As can be seen from the figure, compared to the positive control group (pure solvent) mice, the positive treatment group (Vit+DP+TA treated) mice showed a significant decrease in the time required to find the hidden platform (escape latency) and the distance traveled during the platform concealment phase, as the number of learning days increased. This suggests that Vit+DP+TA treatment can significantly improve the learning and memory functions of mice. During the non-platform phase, the Vit+DP+TA treated mice had a shorter escape latency, spent longer in the platform quadrant, and passed through the original platform position significantly more times than the positive control group (pure solvent) mice. These findings suggest a significant improvement in the memory function of the mice.

[0090] 2. Pathological examination In this animal experiment, a total of six 5xFAD mice and three wild-type littermate mice (C57BL / 6 mice, all female) were used, all 5 months old. The 5xFAD mice were randomly divided into a positive treatment group (N=3), a positive control group (N=3), and a wild-type mouse group (N=3). The positive treatment group received daily oral administration of nerve repair factor (Vit) (see Table 1 for the dosage of each component), while the positive control group received oral administration of a pure solvent (1% Tween 80 + 1% methylcellulose + 98% water, by mass ratio) for a period of 3 months. Subsequently, animal pathology experiments were performed.

[0091] 1. Therapy with nerve repair factors reduces Aβ deposition and microglial cell activation in the hippocampal dentate gyrus (DG) region. In the experiment, 3 μm thick paraffin sections of mouse spinal cord were used and analyzed by immunofluorescence staining. The experimental groups consisted of a wild control group, a positive control group, and a positive treatment group, with three mice used in each group. The tissues were fixed with 4% paraformaldehyde for 24 hours, dehydrated, cleared, and embedded in paraffin. The sections were deparaffinized with xylene, rehydrated stepwise with ethanol, and antigen retrieved with 0.01 M citrate buffer (pH 6.0) at 95°C for 20 minutes.

[0092] For immunofluorescence staining, Iba1 primary antibody and Aβ(6E10) antibody were used and incubated overnight at 4°C. The following day, Alexa Fluor 488-labeled secondary antibody was incubated at room temperature for 2 hours, and the nuclei were stained with DAPI (1:1000) for 10 minutes. Images were acquired using a Zeiss LSM 880 confocal laser microscope with a 20× objective lens. The cell area of ​​each sample was calculated using ImageJ software.

[0093] As shown in Figure 7, the status of Aβ deposition and microglial cell activation in the DG region was compared between wild-type mice (WT), positive control group (pure solvent) mice, and positive treatment group (5xFAD mice treated with nerve repair factors). On the left, Aβ (labeled with 6E10 antibody, green), microglia marker Iba1 (red), DAPI staining (blue), and a composite image of the three are shown in order. WT mice showed almost no Aβ plaques or microglial activation, while positive control group (pure solvent) mice showed significant Aβ plaques (green) and microglial activation (red). After nerve repair factor treatment, both Aβ deposition and microglial activation in the positive treatment group mice were significantly reduced. The bar graph on the right quantifies the Aβ plaque area and Iba1-positive microglia area within the DG region. Compared to the wild-type (WT) group, the Aβ plaque area was significantly increased (****p<0.0001) and the Iba1-positive area was also significantly increased (****p<0.0001) in the positive control group (pine solvent) mice. After treatment with nerve repair factors, both the Aβ plaque area and the Iba1-positive area were significantly decreased in the positive treatment group mice (*p<0.05 or **p<0.01). Scale bar: 50 μm.

[0094] 2. Therapy with nerve repair factors reduces APP and phosphorylated APP deposition in the hippocampal dentate gyrus (DG) region. In the experiment, 3 μm thick paraffin sections of mouse spinal cord were used and analyzed by immunofluorescence staining. The experimental groups consisted of a wild control group, a positive control group, and a positive treatment group, with three mice used in each group. The tissues were fixed with 4% paraformaldehyde for 24 hours, dehydrated, cleared, and embedded in paraffin. The sections were deparaffinized with xylene, rehydrated stepwise with ethanol, and antigen retrieved with 0.01 M citrate buffer (pH 6.0) at 95°C for 20 minutes.

[0095] For immunofluorescence staining, phosphorylated APP (p-APP) primary antibody and APP primary antibody were used and incubated overnight at 4°C. The following day, Alexa Fluor 488-labeled secondary antibody was incubated at room temperature for 2 hours, and the nuclei were stained with DAPI (1:1000) for 10 minutes. Images were acquired using a Zeiss LSM 880 confocal laser microscope with a 20× objective lens. The cell area of ​​each sample was calculated using ImageJ software.

[0096] As shown in Figure 8, the expression of APP and p-APP (Thr668 site) in the DG region was compared in wild-type mice (WT), positive control mice (pure solvent), and positive treatment mice (5xFAD mice treated with nerve repair factors). On the left, APP (green), p-APP (Thr668, red), DAPI staining (blue), and a composite image of the three are shown in order. WT mice showed low expression of both APP and p-APP. Positive control mice (pure solvent) showed a significant increase in APP (green) and p-APP (red) in the DG region, indicating accumulation of pathological APP and p-APP. After nerve repair factor treatment, both APP and p-APP expression in positive treatment mice were clearly reduced. The bar graph on the right quantifies the p-APP (Thr668) positive area in the DG region. Compared to the wild-type (WT) group, p-APP was significantly increased in the positive control group (pure solvent) mice (***p<0.001), while the p-APP area was significantly decreased in the positive treatment group mice after nerve repair factor therapy (**p<0.01). Scale bar: 50 μm.

[0097] 3. Nerve repair factor therapy reduces APP and Tau protein levels. The expression and phosphorylation levels of APP and Tau proteins in mouse brain tissue were measured by Western blotting. First, protein samples were extracted from the brain tissue of three groups of mice: wild-type control, positive control, and positive treatment group. After protein quantification by BCA, equal volumes of samples were subjected to electrophoresis on an SDS-PAGE gel and then transferred to a PVDF membrane. After blocking with 5% skim milk powder, anti-p-APP (Thr668), APP, p-Tau (AT180) (Thr231), Tau5, and GAPDH (internal reference) antibodies were incubated overnight at 4°C. The following day, HRP-labeled secondary antibodies were incubated at room temperature for 1-2 hours.

[0098] After chemiluminescence detection using an ECL substrate, grayscale analysis of the bands was performed using ImageJ software to calculate the relative expression levels of the target protein and GAPDH.

[0099] As shown in Figure 9, the expression analysis of p-APP(Thr668), APP, p-Tau(AT180), and Tau5 proteins in wild-type mice (WT), positive control (pure solvent) mice, and positive treatment group (5xFAD mice treated with nerve repair factors) is shown by Western blotting, with GAPDH as the internal reference. Compared to WT mice, the expression levels of p-APP(Thr668) and p-Tau(AT180) were significantly increased in the positive control (pure solvent) mice, while the expression levels of both p-APP(Thr668) and p-Tau(AT180) were decreased in the positive treatment group mice after nerve repair factor treatment. The quantitative analysis bar graphs show the ratios of p-APP / APP and p-Tau / Tau5. Compared to the WT group, the p-APP / APP ratio was significantly increased in the positive control (pure solvent) mice (**p<0.01), and this ratio was significantly decreased after nerve repair factor treatment (*p<0.05). The p-Tau / Tau5 ratio in the positive control group (pure solvent) mice also significantly increased (*p<0.05), and this ratio significantly decreased after treatment with nerve repair factors (*p<0.05).

[0100] 4. Nerve repair factor therapy improves the secretion of Reelin protein in the mouse hippocampal dentate gyrus. In the experiment, 3 μm thick paraffin sections of mouse spinal cord were used and analyzed by immunofluorescence staining. The experimental groups consisted of a wild control group, a positive control group, and a positive treatment group, with three mice used in each group. The tissues were fixed with 4% paraformaldehyde for 24 hours, dehydrated, cleared, and embedded in paraffin. The sections were deparaffinized with xylene, rehydrated stepwise with ethanol, and antigen retrieved with 0.01 M citrate buffer (pH 6.0) at 95°C for 20 minutes.

[0101] For immunofluorescence staining, Reelin primary antibody was used and incubated overnight at 4°C. The following day, Alexa Fluor 488-labeled secondary antibody was incubated at room temperature for 2 hours, followed by nuclear staining with DAPI (1:1000) for 10 minutes. Images were acquired using a Zeiss LSM 880 confocal laser microscope with a 20× objective lens. The cell area of ​​each sample was calculated using ImageJ software.

[0102] As shown in Figure 10, the expression of Reelin protein in the DG region of wild-type mice (WT), positive control (pure solvent) mice, and positive treatment group (5xFAD mice treated with nerve repair factors) is shown. From left to right, the images are Reelin (green), DAPI (blue), and a composite image of both. WT mice showed abundant Reelin expression in the DG region. Positive control (pure solvent) mice showed significantly reduced Reelin expression. After nerve repair factor treatment, Reelin expression in positive treatment group mice recovered significantly. The bar graph on the right shows a quantitative analysis of the Reelin-positive area in the DG region. Compared to the WT group, Reelin expression in positive control (pure solvent) mice was significantly reduced (*p<0.05), and after nerve repair factor treatment, Reelin expression significantly increased (*p<0.05). Scale bar: 50 μm.

[0103] The present invention has been described in detail above. Those skilled in the art should understand that the present invention can be widely implemented under equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without unnecessary experimentation. While specific embodiments are provided, it should be understood that further improvements to the invention are possible. In short, in accordance with the principles of the invention, this application is intended to encompass modifications, uses, or improvements using prior art known in the art, even if they deviate from the scope already disclosed herein.

Claims

1. (A1) Manufacturing of products for the treatment and / or prevention of neurodegenerative diseases, (A2) A composition for improving cognitive function used in the manufacture of products for improving symptoms of neurodegenerative diseases, (d1) Docosahexaenoic acid: 8,000 to 24,000 parts by mass Phosphatidylserine: 2400 to 7200 parts by mass Vitamin D3: 1 part by mass Vitamin B12: 0.5 to 2 parts by mass Vitamin B6: 50-150 parts by mass Vitamin B1: 100-300 parts by mass Vitamin B9: 4-12 parts by mass Taurine: 2500-7500 parts by mass Astaxanthin: A composition comprising 500 to 1500 parts by mass of docosahexaenoic acid, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and astaxanthin, or (d2) Docosahexaenoic acid: 0.5 to 2 moles Phosphatidylserine: 0.5 to 2 moles Vitamin D3: 1 mole Vitamin B12: 0.5 to 2 moles Vitamin B6: 0.5 to 2 moles Vitamin B1: 0.5 to 2 moles Vitamin B9: 0.5 to 2 moles Taurine: 0.5 to 2 moles A composition characterized by being one of the following: astaxanthin, 0.5 to 2 moles of docosahexaenoic acid, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine, and astaxanthin.

2. In (d1) above, the mass of each component is Docosahexaenoic acid: 16,000 parts by mass Phosphatidylserine: 4800 parts by mass Vitamin D3: 1 part by mass Vitamin B12: 1 part by mass Vitamin B6: 100 parts by mass Vitamin B1: 200 parts by mass Vitamin B9: 8 parts by mass Taurine: 5000 parts by mass The composition according to claim 1, characterized in that it comprises 1,000 parts by mass of astaxanthin.

3. In (d2) above, the molar portion of each component is, Docosahexaenoic acid: 1 mole Phosphatidylserine: 1 mole Vitamin D3: 1 mole Vitamin B12: 1 mole Vitamin B6: 1 mole Vitamin B1: 1 mole Vitamin B9: 1 mole Taurine: 0.5 to 2 moles The composition according to claim 1, characterized in that it contains astaxanthin: 1 mole part.

4. Use of the composition according to any one of claims 1 to 3 in the manufacture of a product for reducing toxicity to nerve cells by neurotoxic proteins.

5. The use according to claim 4, characterized in that the nerve cells are primary nerve cells, SH-SY5Y cells, or iPSC pluripotent stem cell-induced nerve cells.

6. The composition comprises the composition according to any one of claims 1 to 3, (B1) Treatment and / or prevention of neurodegenerative diseases (B2) Improvement of symptoms of neurodegenerative diseases (B3) A product characterized by being used in any of the following ways: to reduce the toxic effects on nerve cells caused by neurotoxic proteins.

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