Method for preparing composition capsule containing sea squirt extract, DHA and NMN

Abstract

The invention provides a method for preparing a composition capsule containing a sea squirt extract, DHA and NMN, and relates to the technical field of health-care food. The composition capsule is composed of the sea squirt extract, free docosahexaenoic acid, nicotinamide mononucleotide and vitamin E. The composition comprises the following components in parts by weight: 14-20 parts of the sea squirt extract, 20-40 parts of the free docosahexaenoic acid, 40-53 parts of the nicotinamide mononucleotide and 0.2-2 parts of the vitamin E. The plasmalogen in the sea squirt extract can improve the function of a cell membrane in brain cells and improve the immunity of the brain cells. DHA has the effect of promoting formation and development of brain cells, NMN is utilized for promoting synthesis and release of neurotransmitters to improve the function stability of a nervous system, vitamin E serves as an antioxidant and an anti-inflammatory agent to enhance nerve protection, and the composition capsule has the synergistic effect, so that the brain function is remarkably enhanced, brain aging is delayed, and the effect of improving the brain function is achieved. The senile dementia is effectively prevented and improved.

Description

Technical Field

[0001] The present invention relates to the technical field of health foods, and particularly to a method for preparing a composition capsule containing ascidian extract, DHA and NMN. Background Art

[0002] With the accelerating aging of the population and the increase in average life expectancy, the number of Alzheimer's disease patients is gradually increasing. Alzheimer's disease is increasingly becoming a serious social problem and has become one of the most serious diseases threatening the lives and health of the elderly in China. With the increase in age, the prevalence of Alzheimer's disease gradually rises. Alzheimer's disease is a progressive neurodegenerative disease, and primary degenerative dementia and vascular dementia are important conditions of Alzheimer's disease. Currently, there is no ideal treatment plan for the middle and late stages of Alzheimer's disease in medical technology. Therefore, the prevention and treatment of Alzheimer's disease in the early stage are of great significance.

[0003] According to an antioxidant composition containing NMN and its preparation method disclosed in Chinese Patent Publication No. "CN114272273A", it is prepared from raw materials containing the following parts by mass: 10-13 parts of nicotinamide mononucleotide, 0.02-0.06 part of coenzyme Q10, 0.2-0.5 part of plasmalogen, 0.8-2.0 parts of nucleic acid, 0.05-1 part of amino acid, 0.2-0.5 part of vitamin, and 0.005-0.08 part of trace element. The antioxidant composition provided by the present invention adopts specific components and specific ratios, with reasonable formulation and proper compatibility, and has the advantages of good antioxidant and anti-aging effects, no harm to the human body, and stable product quality. It is convenient to use, has good absorption effect, and no adverse and toxic side effects.

[0004] The above patent documents and the prior art have the following technical problems in use:

[0005] Problem 1: Many health foods or drugs for Alzheimer's disease rely on a single active ingredient (DHA or antioxidant), with a single mechanism of action, making it difficult to simultaneously solve multiple pathological problems such as nerve cell membrane damage, insufficient brain cell development, neurotransmitter imbalance, and oxidative stress, resulting in insufficient comprehensive enhancement of brain function and limited effects in preventing and treating Alzheimer's disease.

[0006] Problem 2: The prior art usually extracts plasmalogen from fish oil, meat (salmon, chicken), etc. The plasmalogen content from these sources is low, the purification is difficult, and its endogenous antioxidant properties are not fully utilized, resulting in limited effects in the treatment of Alzheimer's disease and being unable to effectively support the repair and antioxidant needs of brain cells.

[0007] Problem 3: Most of the existing health products or drug formulas have fixed ratios and cannot be adjusted according to the individual differences (gene background) of patients. Moreover, they lack the comprehensive support of multi-phospholipid synergy, antioxidant, and microcirculation improvement, resulting in poor efficacy in specific populations, limited scope of application, and inability to meet the needs of early prevention and early treatment of Alzheimer's disease simultaneously. Summary of the Invention

[0008] Technical Problems to be Solved

[0009] In view of the deficiencies of the prior art, the present invention provides a method for preparing a composition capsule containing ascidian extract, DHA, and NMN, and solves the following problems:

[0010] 1. Aiming at the problem that the mechanism of action of a single component in the technology is limited and it is difficult to comprehensively address the multiple pathological characteristics of Alzheimer's disease;

[0011] 2. Aiming at the problem that the extraction source of traditional plasmalogens has low content and poor purity, which limits their application effect in the treatment of Alzheimer's disease;

[0012] 3. Aiming at the problem that the existing treatment of Alzheimer's disease lacks personalized design and multi-functional support, has a narrow scope of application, and weak efficacy targeting.

[0013] Technical Solution

[0014] To achieve the above objectives, the present invention is achieved through the following technical solutions: A method for preparing a composition capsule containing ascidian extract, DHA, and NMN, the composition capsule is composed of ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E, and the preparation method of the composition capsule is as follows:

[0015] Sp1: Ascidian extract extraction: Using ascidian as the extraction raw material, a mixed solvent of an alcohol entrainer and supercritical CO2 is used to remove the grease and neutral fat in the ascidian, and a mixed solvent of an alcohol entrainer and supercritical CO2 is used to extract phospholipid active substances with relatively high polarity to obtain ascidian extract. The extraction environment is 40°C and 300 bar;

[0016] Sp2: Preparation of the content: Mix the ascidian extract, free DHA, NMN, and vitamin E to obtain a preliminary mixed content, then use a colloid mill to perform three homogenization treatments on the preliminary mixed content, and then place the homogenized preliminary mixed content in a stirring tank and stir for 40 - 60 min, and then perform filtration treatment to obtain a content with uniform components for standby;

[0017] Sp3: Sol preparation: Add glycerol and purified water to a gel-making tank, heat to 70 - 80°C, then add gelatin, stir while adding, and perform heat preservation treatment until all the gelatin is dissolved to obtain a mixed gel solution;

[0018] Sp4: Vacuum extraction and degassing: Place the mixed adhesive solution under vacuum conditions until no bubbles are generated on the surface of the mixed adhesive solution. After adjusting the viscosity of the mixed adhesive solution, filter the mixed adhesive solution. Place the filtered mixed adhesive solution in a heat preservation barrel and let it stand at a temperature of 65°C ± 8°C for not less than 2 hours to obtain the capsule material.

[0019] Sp5: After pelletizing, shaping, drying, selecting pellets and packaging the capsule material obtained in step Sp4 and the content obtained in step Sp2, fish oil soft capsules are obtained.

[0020] Preferably, the component combination of the sea squirt extract, free docosahexaenoic acid, nicotinamide mononucleotide and vitamin E in the composition capsule is: 14 - 20 parts of sea squirt extract, 20 - 40 parts of free docosahexaenoic acid, 40 - 53 parts of nicotinamide mononucleotide and 0.2 - 2 parts of vitamin E, and the total component combination of the composition capsule is 100 parts.

[0021] Preferably, when extracting the sea squirt extract, the extraction time is 2 - 4 hours, the CO2 flow rate is 10 - 15 L / min, 5% - 10% ethanol is added as a co-solvent during the extraction process, the content of plasmalogen in the obtained sea squirt extract is not less than 1%, the extraction pressure is 10 - 45 MPa, and the extraction temperature is 30 - 50°C.

[0022] Preferably, the raw material of the free docosahexaenoic acid is fish oil, which is refined by supercritical fluid extraction technology to remove heavy metals and impurities.

[0023] Preferably, lecithin and cephalin are further added as auxiliary phospholipid components in the composition capsule, where the content of lecithin is 10% - 20% of the sea squirt extract, and the content of cephalin is 5% - 10% of the sea squirt extract.

[0024] Preferably, ginkgo biloba extract is added as an additional neuroprotective component in the composition capsule, and the content of ginkgo biloba extract is 5% - 10% of the total weight of the composition.

[0025] Preferably, prebiotics are added as enteric-brain axis regulating components in the composition capsule, and the content of prebiotics is 10% - 15% of the total weight of the composition.

[0026] Preferably, the ratio range of the sea squirt extract to the free docosahexaenoic acid in the combination capsule is between 1:3 and 1:1.

[0027] Beneficial effects

[0028] The present invention provides a method for preparing a composition capsule containing a sea squirt extract, DHA and NMN.

[0029] It has the following beneficial effects:

[0030] 1. The present invention uses ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E as the main raw materials to prepare a composition capsule. The plasmalogens in the ascidian extract improve the cell membrane function in brain cells. DHA plays a role in promoting the formation and development of brain cells. NMN is used to promote the synthesis and release of neurotransmitters to improve the functional stability of the nervous system. As an antioxidant and anti-inflammatory agent, vitamin E enhances neuroprotection. The four components of the entire composition capsule act synergistically to significantly enhance brain function, delay brain aging, and effectively prevent and improve Alzheimer's disease.

[0031] 2. The present invention changes the traditional method of extracting plasmalogens from fish oil and meat, and selects ascidians with higher content and purity of plasmalogens. At the same time, plasmalogens are used for the prevention of Alzheimer's disease. Since the vinyl ether bond at the sn-1 position of plasmalogens is vulnerable to free radical attack, it can act as an effective endogenous antioxidant to protect easily oxidized lipids and lipoproteins. At the same time, because plasmalogens are distributed on the cell membrane and participate in various physiological activities of cells, it can reduce the effects leading to central nervous system diseases, promote the growth and recovery of brain cells in the elderly, and prevent brain degeneration caused by the increase in the age of the elderly, thus effectively preventing Alzheimer's disease and improving memory.

[0032] 3. The present invention uses ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E as the main raw materials to prepare a composition capsule, and adjusts the ratio according to the genetic background and symptoms of actual patients. When the entire composition capsule is prepared, in addition to the main components, lecithin, cephalin, and plasmalogens are added to act synergistically, enhancing the repair and protection ability of the nerve cell membrane. The combination of different phospholipid types provides more comprehensive lipid support, improving the prevention and treatment effect of the formula on neurodegenerative diseases. At the same time, the combined action of natural ginkgo biloba extract and vitamin E provides dual antioxidant protection, reducing the damage of oxidative stress to brain cells, and improving cerebral microcirculation at the same time, significantly enhancing the neuroprotective efficacy of the capsule, realizing the multi-functional adaptation use and targeted ratio of the entire composition capsule formula, and carrying out the early prevention and early treatment of Alzheimer's symptoms, so as to achieve the effects of cost savings and wider application scope. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a structural diagram of the composition capsule formula of the present invention;

[0034] Figure 2 It is a preparation method diagram of the composition capsule of the present invention;

[0035] Figure 3 It is a content table of plasmalogens in each raw material of the present invention;

[0036] Figure 4 Bar chart of the content of plasmalogens in each raw material of the present invention;

[0037] Figure 5 Comparison chart of the combination of plasmalogens of the present invention with other components. Detailed implementation manners

[0038] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention. Specific embodiment 1:

[0040] As Figures 1 to 5 shown, a method for preparing a composition capsule containing ascidian extract, DHA, and NMN, the composition capsule is composed of ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E, and the components of ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E in the composition capsule are: 14-20 parts of ascidian extract, 20-40 parts of free docosahexaenoic acid, 40-53 parts of nicotinamide mononucleotide, and 0.2-2 parts of vitamin E. The ratio range of ascidian extract to free docosahexaenoic acid in the combined capsule is between 1:3 and 1:1. The preparation method of the composition capsule is as follows:

[0041] Sp1: Ascidian extract: Using ascidian as the extraction raw material, a mixed solvent of alcohol entrainer and supercritical CO2 is used to remove the oils and neutral fats in the ascidian, and a mixed solvent of alcohol entrainer and supercritical CO2 is used to extract the phospholipid active substances with relatively large polarity to obtain the ascidian extract. The extraction environment is 40°C, 300 bar, the extraction time is 2-4 h, the CO2 flow rate is 10-15 L / min, 5%-10% ethanol is added as a cosolvent during the extraction process, the content in the obtained ascidian extract is not less than 30%, the extraction pressure is 10-45 MPa, and the extraction temperature is 30-50°C;

[0042] Sp2: Preparation of the content: Mix the ascidian extract, free DHA, NMN, and vitamin E to obtain a preliminary mixed content, then use a colloid mill to perform three homogenization treatments on the preliminary mixed content, and then place the homogenized preliminary mixed content in a stirring tank and stir for 40-60 min, and then perform a filtration treatment to obtain a content with uniform composition for standby;

[0043] Sp3: Gel sol: Add glycerol and purified water to a gel melting tank, heat to 70-80°C, then add gelatin, stir while adding, and perform heat preservation treatment until all the gelatin is dissolved to obtain a mixed gel solution;

[0044] Sp4: Vacuum degassing: Place the mixed adhesive solution under vacuum conditions until no bubbles are generated on the surface of the mixed adhesive solution. After adjusting the viscosity of the mixed adhesive solution, filter the mixed adhesive solution. Place the filtered mixed adhesive solution in a heat preservation bucket and let it stand at a temperature of 65°C ± 8°C for not less than 2 hours to obtain the capsule material.

[0045] Sp5: After pelletizing, shaping, drying, pill selection, and packaging of the capsule material obtained in step Sp4 and the content obtained in step Sp2, soft capsules are obtained.

[0046] For the preparation method of the above composition capsules, during actual preparation, the actual weight ratios of the respective components of the generally selected composition capsules are as follows:

[0047] In the composition capsules: 90 mg of ascidian extract, 130 mg of free docosahexaenoic acid, 250 mg of β-nicotinamide mononucleotide, and 5 mg of vitamin E. The weight of the capsule cap and capsule body is 94 mg, and the total weight of the composition capsules is 570 ± 5% mg. The specification of the capsules used in this ratio is size 0 capsules of relevant national standards, which is the standard ratio. During actual use, the content of each component can be adjusted correspondingly according to the corresponding ratio in combination with the patient's case or the requirements of the production capsule model. For example, size 1 capsules are smaller in size than size 0 capsules, and the amount of powder they can hold is relatively reduced. Therefore, during preparation, the amount of the corresponding components is adjusted proportionally. The free docosahexaenoic acid uses high-concentration Omega-3 DHA fish oil from German KDPharma, which is refined by supercritical fluid extraction technology to remove heavy metals and impurities. The composition capsules are prepared using ascidian extract, free docosahexaenoic acid, nicotinamide mononucleotide, and vitamin E as the main raw materials. The plasmalogens extracted from ascidians in the composition capsules improve the cell membrane function in brain cells. DHA plays a role in promoting the formation and development of brain cells. NMN is used to promote the synthesis and release of neurotransmitters to improve the functional stability of the nervous system. As an antioxidant and anti-inflammatory agent, vitamin E enhances neuroprotection. The four components of the entire composition capsule work synergistically to significantly enhance brain function, delay brain aging, effectively prevent and improve Alzheimer's disease. At the same time, it changes the traditional way of extracting plasmalogens from fish oil and meat, selects ascidians with higher content and purity of plasmalogens, and uses plasmalogens for the treatment of Alzheimer's disease. Since the vinyl ether bond at the sn-1 position of plasmalogens is vulnerable to free radical attack, it can achieve the effect of an effective endogenous antioxidant, protecting easily oxidized lipids and lipoproteins. At the same time, because plasmalogens are distributed on the cell membrane and participate in various physiological activities of cells, it can reduce the effects leading to central nervous system diseases, promote the growth and recovery of brain cells in the elderly, and prevent brain degeneration caused by the aging of the elderly, thereby effectively preventing Alzheimer's disease and improving memory.

[0048] To increase the personalization of the entire composition capsule for treatment and expand the scope of adaptable treatment, during actual use, lecithin and cephalin are further added to the composition capsule as auxiliary phospholipid components. The content of lecithin is 10%-20% of the ascidian extract, and the content of cephalin is 5%-10% of the ascidian extract, so as to enhance the synergistic effect among various phospholipids and improve the neuroprotective effect. Lecithin and cephalin are further added to the composition capsule as auxiliary phospholipid components. The content of lecithin is 10%-20% of the ascidian extract (if the ascidian extract is 100 mg, then lecithin is 10-20 mg), and the content of cephalin is 5%-10% of the ascidian extract (if the ascidian extract is 100 mg, then cephalin is 5-10 mg), aiming to enhance the synergistic effect among various phospholipids and improve the neuroprotective effect; Lecithin (mainly phosphatidylcholine) supports cell membrane fluidity and acetylcholine synthesis, and synergistically maintains membrane stability with the antioxidant properties of plasmalogens; Cephalin (mainly phosphatidylethanolamine and phosphatidylserine) supports membrane fusion and neuron survival, and jointly resists oxidative stress with plasmalogens to provide comprehensive lipid support, which may improve brain function and delay cognitive decline.

[0049] Ginkgo biloba extract is added to the composition capsule as an additional neuroprotective component. The content of Ginkgo biloba extract is 5%-10% of the total weight of the composition, so as to provide additional antioxidant effects and improve cerebral blood flow. Ginkgo biloba extract is added as an additional neuroprotective component, with a content of 5%-10% of the total weight of the composition (if the total weight is 570 mg, then it is 28.5-57 mg). It scavenges free radicals through the antioxidant effects of flavonoids and terpenoids, reduces oxidative stress, forms a double protection with vitamin E, and improves cerebral blood flow by dilating blood vessels, promotes the delivery of oxygen and nutrients, and synergistically enhances the neuronal environment with DHA and plasmalogens, which may significantly improve the curative effect, especially suitable for patients with cerebrovascular problems.

[0050] After the capsule particles are encapsulated, the microcapsule technology is added, and microcapsules with controlled release function are formed through the spray drying process. The release time of the microcapsules is adjusted to 4-12 h according to needs, so as to achieve the sustained release of active ingredients. The microcapsules are gradually decomposed in the gastrointestinal tract to ensure the sustained release of easily degradable components such as DHA and NMN into the blood and brain, improve the bioavailability and medication compliance, and are especially suitable for elderly patients. Research shows that the controlled release technology can improve the drug efficacy.

[0051] Prebiotics are added to the composition capsules as gut-brain axis regulating components, and the content of prebiotics is 10%-15% of the total weight of the composition to enhance brain health and cognitive function by regulating the gut microbiota. Prebiotics are added as gut-brain axis regulating components, with a content of 10%-15% of the total weight (if the total weight is 570 mg, then 57-85.5 mg), such as fructooligosaccharides or inulin, which promote the growth of beneficial bacteria, improve the gut barrier, reduce inflammatory metabolites, regulate neuroinflammation and neurotransmitter synthesis through short-chain fatty acids, enhance energy metabolism with NMN, enhance absorption efficiency with plasmalogens and DHA, and indirectly improve cognitive function. Research supports its association with brain health.

[0052] These improvements enable the capsules to enhance neuroprotection and adaptability through phospholipid synergy, antioxidant, controlled release, and gut-brain axis regulation, and are suitable for patients with mild cognitive impairment, Alzheimer's disease with vascular problems, poor medication compliance, or gut disorders. However, further clinical trials are needed to verify the long-term effects and optimal dosage. Lecithin and cephalin provide diverse lipid support and strengthen cell membrane protection; Ginkgo biloba extract supplements antioxidant and blood flow improvement effects; microcapsule technology achieves controlled release and improves the persistence of efficacy; prebiotics support brain health through the gut-brain axis, significantly enhancing the versatility and specificity of the composition capsules, making them more suitable for personalized treatment and meeting different needs.

[0053] The ratio of the composition capsules is personalized according to the individual's genetic background or metabolic characteristics. For carriers of the APOE4 genotype, the proportion of free DHA is increased to 40-45 parts, and the proportion of NMN is adjusted to 55-60 parts to optimize the cognitive enhancement effect for specific populations. And the ratio is adjusted according to the actual patient's genetic background and symptoms. When preparing the whole composition capsules, in addition to the main components, lecithin, cephalin, and plasmalogens are added to act synergistically, enhancing the repair and protection ability of the nerve cell membrane. The combination of different phospholipid types provides more comprehensive lipid support and improves the prevention and treatment effect of the formula on neurodegenerative diseases. At the same time, the combined action of natural Ginkgo biloba extract and vitamin E provides dual antioxidant protection, reduces the damage of oxidative stress to brain cells, and improves cerebral microcirculation, significantly enhancing the neuroprotective efficacy of the capsules, realizing the multi-functional adaptation use and targeted ratio of the whole composition capsule formula, and conducting early prevention and early treatment of Alzheimer's symptoms, thus achieving the effects of cost savings and increased scope of application. Specific Example 2:

[0055] As Figures 1 to 5 shown, based on the content in the above specific examples, the following content is further disclosed:

[0056] To verify the interaction and irreplaceability of each component in the composition capsule, this application conducts a control experiment. By comparing the effects of the complete formula and the formula lacking a single ingredient on cognitive function, and at the same time comparing the effects of the components in the formula after being replaced, a control experiment is carried out. The experimental content is as follows:

[0057] The following experiment is set according to the actual weight in the proportion of 14 - 20 parts of the ascidian extract, 20 - 40 parts of free docosahexaenoic acid, 40 - 53 parts of nicotinamide mononucleotide, and 0.2 - 2 parts of vitamin E as described above. The following experiment uses capsules with adjusted total amounts. The specific weights are: 45 mg of ascidian extract, 65 mg of free docosahexaenoic acid, 125 mg of β-nicotinamide mononucleotide, and 2.5 mg of vitamin E;

[0058] Experimental design: Use the control group and the experimental group to remove the ascidian extract, DHA, NMN, or vitamin E respectively, and compare the cognitive function, oxidative stress, and neuroprotective effects;

[0059] Experimental content: The APP / PS1 transgenic mice are selected as the animal model. The experimental grouping is shown in Table 1 below:

[0060]

[0061] Table 1

[0062] Administration: Oral administration is given daily for 3 months;

[0063] Measurement: Morris water maze, fear conditioning, novel object recognition test are selected; Biomarkers include amyloid-β level in the brain, tau protein phosphorylation, oxidative stress marker (MDA), inflammatory markers (TNF-α, IL-6), NAD+ level;

[0064] Result analysis: Analyze according to the measurement experiment. The analysis results are shown in the following table:

[0065] The results of the behavioral tests are shown in Tables 2, 3, and 4:

[0066] The results of the Morris water maze test are shown in Table 2 below: In the pre-escape period (Day 5, seconds):

[0067]

[0068] Table 2

[0069] In Table 2 above, the escape latency of the complete group on the 5th day is 10 ± 2 seconds, which is significantly lower than that of the control group (25 ± 3 seconds, p < 0.001). After removing any component or replacing it, the escape latency is prolonged. For example, the escape latency of the group without DHA is 18 ± 3 seconds, and the group without NMN is 16 ± 2 seconds.

[0070] The results of the fear conditioning test are shown in Table 3 below:

[0071]

[0072]

[0073] Table 3

[0074] In Table 3 above, the freezing time of the complete group in the context test was 60 ± 6%, significantly higher than that of the control group (30 ± 5%, p < 0.01). After removing the components, the freezing time decreased, such as 45 ± 5% for the group without the ascidian extract.

[0075] The results of the novel object recognition test are shown in Table 4 below:

[0076]

[0077]

[0078] Table 4

[0079] In Table 4 above, the discrimination index of the complete group was 0.6 ± 0.06, significantly higher than that of the control group (0.2 ± 0.05, p < 0.001). After removing or replacing the components, the discrimination index decreased, such as 0.25 ± 0.03 for the group replacing DHA with EPA.

[0080] The biomarker results are shown in Tables 5, 6, and 7:

[0081] The levels of amyloid-β (Aβ) and phosphorylated tau protein are shown in Table 5 below:

[0082]

[0083] Table 5

[0084] The levels of oxidative stress and inflammation markers are shown in Table 6:

[0085]

[0086]

[0087] The NAD+ levels in Table 6 are shown in Table 7:

[0088]

[0089]

[0090] Table 7

[0091] According to the above biochemical biomarker results, the complete group performed best in all biochemical biomarkers:

[0092] Amyloid-β level: 200 ± 20 pg / mg protein, significantly lower than that of the control group (500 ± 50, p < 0.001);

[0093] Phosphorylation of tau protein: 0.3 ± 0.05, significantly lower than that of the control group (0.8 ± 0.1, p < 0.01);

[0094] MDA level: 2 ± 0.2 nmol / mg protein, significantly lower than that of the control group (5 ± 0.5, p < 0.001);

[0095] TNF-α level: 40 ± 4 pg / mg protein, significantly lower than that of the control group (100 ± 10, p < 0.01);

[0096] IL-6 level: 15 ± 1.5 pg / mg protein, significantly lower than that of the control group (50 ± 5, p < 0.001);

[0097] NAD+ level: 200 ± 20 nmol / mg protein, significantly higher than that of the control group (100 ± 10, p < 0.01);

[0098] After removing or replacing the components, the levels of each biomarker deteriorated. For example, in the group without NMN, the NAD+ level decreased to 120 ± 12;

[0099] A notable finding is that after replacing NMN with nicotinamide, although the NAD+ level was lower than that of the complete group, it was higher than that of the group without NMN, suggesting that nicotinamide may partially compensate for the role of NMN, but the effect is not as good as direct supplementation of NMN.

[0100] Result analysis: The complete capsule group performed the best. In all behavioral tests, the complete capsule group showed the shortest escape latency, the highest freezing time, and discrimination index, indicating that it significantly improved spatial memory, fear memory, and novel object recognition ability; among the biochemical markers, the Aβ level, tau phosphorylation, oxidative stress, and inflammatory markers of the complete group were the lowest, and the NAD+ level was the highest, showing its excellent effects in neuroprotection and anti-aging.

[0101] The results of the component removal effect are as follows:

[0102] Group without ascidian extract: The Aβ level and tau phosphorylation increased, indicating impaired nerve cell membrane function;

[0103] Group without free DHA: The decline in cognitive function was the most obvious (escape latency 20 ± 4 seconds), and the Aβ level increased significantly, suggesting the irreplaceability of DHA for brain cell development;

[0104] Group without NMN: The NAD+ level was the lowest (120 ± 12), and cognitive function and neurotransmitter synthesis were impaired;

[0105] Group without vitamin E: Slightly elevated oxidative stress and inflammation markers, weakened antioxidant protection.

[0106] The results of the component replacement effect are as follows:

[0107] Replacing the ascidian extract (soybean lecithin): The effect is inferior to the complete group, and the Aβ level rises to 350 ± 35, indicating that the unique antioxidant effect of the ascidian extract is irreplaceable;

[0108] Replacing free DHA (EPA): The cognitive function is the worst (discrimination index 0.25 ± 0.03), suggesting that the specific effect of DHA is superior to other fatty acids;

[0109] Replacing NMN (nicotinamide): The NAD+ level (150 ± 15) is higher than the group without NMN but lower than the complete group, indicating that nicotinamide partially compensates but the effect is not as good as NMN;

[0110] Replacing vitamin E: The antioxidant and anti-inflammatory effects are weakened, and the IL-6 level rises to 28 ± 2.8.

[0111] Verification of synergistic effect: Removing or replacing any one component leads to a decrease in effect, proving that the synergistic effect of ascidian extract, DHA, NMN, and vitamin E is crucial;

[0112] Experimental conclusion: The tabular data clearly show that the complete formula capsule has significantly better effects in cognitive function, neuroprotection, and anti-aging than the control group and other experimental groups. Removing or replacing any component leads to a decrease in curative effect, verifying the irreplaceability of each component and its synergistic effect, providing strong experimental evidence for the application of this composition capsule in the prevention and improvement of Alzheimer's disease. Specific Example 3:

[0114] As Figures 1 to 5 shown, based on the content in the above specific examples, the following content is further disclosed:

[0115] To verify the rationality of the proportions of the various components in the composition capsule, a countermeasure research experiment on dosage is conducted. By adjusting the amounts of components DHA, NMN, and vitamin E, the maximized effect on cognitive function is evaluated;

[0116] Experimental content: APP / PS1 transgenic mice are used, and the experimental groups are as shown in Table 8 below:

[0117]

[0118] Table 8

[0119] Administration: Oral administration daily for 3 months;

[0120] Measurements: Morris water maze, fear conditioning, novel object recognition test were selected; Biomarkers include amyloid-β level in the brain, tau protein phosphorylation, oxidative stress markers (MDA), inflammatory markers (TNF-α, IL-6), NAD+ level;

[0121] Personalized experiment: Wild-type mice and APOE knockout mice were used, and standard capsules were given to observe the differences in responses; The ratio was adjusted according to the differences (DHA was increased to 150 mg for APOE knockout mice), and then the effects were tested;

[0122] The experimental results are shown in Tables 9, 10, 11, and 12 as follows:

[0123] The results of the Morris water maze test in the behavioral test are shown in Table 9 as follows:

[0124]

[0125]

[0126] The results of the amyloid-β (Aβ) level of the biochemical marker are shown in Table 10 as follows:

[0127]

[0128]

[0129] Table 10

[0130] The NAD+ level of the biochemical marker (only the NMN dose group) is shown in Table 11 as follows:

[0131]

[0132] Table 11

[0133] The level of the oxidative stress marker MDA of the biochemical marker (only the vitamin E dose group) is shown in Table 12 as follows:

[0134]

[0135]

[0136] Table 12

[0137] The level of the inflammatory marker TNF-α of the biochemical marker (only the vitamin E dose group) is shown in Table 13 as follows:

[0138]

[0139] Table 13

[0140] The results of the personalized experiment are shown in Table 14 as follows:

[0141]

[0142] Table 14

[0143] Result analysis:

[0144] DHA dose group: Optimal dose: 65 - 75 mg, shortest escape latency (9 - 10 s), lowest Aβ level (165 - 200 pg / mg protein). The effect is slightly reduced at 100 mg, probably due to the excessive metabolic burden caused by overdose; 65 mg is the standard ratio, and 75 mg is better under specific circumstances (APOE knockout);

[0145] NMN dose group: Optimal dose: 125 - 150 mg, highest NAD+ level (200 - 220 nmol / mg protein), shortest escape latency (9 - 10 s). The effect is not further improved at 200 mg; 125 mg is the standard ratio, and 150 mg may be the upper limit;

[0146] Vitamin E dose group: Optimal dose: 2.5 - 3 mg, lowest MDA (2.2 - 2.5 nmol / mg protein) and TNF-α (35 - 40 pg / mg protein). No additional benefit is seen at 3.5 mg; 2.5 mg is the standard ratio, and 3 mg may be slightly better but the difference is not significant;

[0147] Ascidian extract dose group: Optimal dose: 45 - 60 mg, shortest escape latency (9 - 10 s), lowest Aβ level (190 - 200 pg / mg protein). The effect is insufficient at 30 mg and slightly reduced at 80 mg; 45 mg is the standard ratio, and 60 mg can be used as an alternative;

[0148] Personalized experiment: Wild-type mice: The standard capsule (45 mg of ascidian extract, 65 mg of free docosahexaenoic acid, 125 mg of β-nicotinamide mononucleotide, and 2.5 mg of vitamin E) has the best effect; APOE knockout mice: The effect of the standard capsule is poor. Increasing DHA to 100 mg significantly improves the effect, approaching the wild-type level, indicating that the genetic background affects the optimal ratio;

[0149] Experimental conclusion: The tabular data shows that the standard capsule: 45 mg of ascidian extract, 65 mg of free docosahexaenoic acid, 125 mg of β-nicotinamide mononucleotide, and 2.5 mg of vitamin E is reasonable and close to the optimal range in most cases:

[0150] DHA: Optimal at 65 - 75 mg, 65 mg as the standard;

[0151] NMN: Optimal at 125 - 150 mg, 125 mg as the standard;

[0152] Vitamin E: The optimal amount is 2.5 - 3 mg, with 225 g as the standard;

[0153] Ascidian extract: The optimal amount is 45 - 60 mg, with 45 mg as the standard.

[0154] Personalized experiment tips: For APOE knockout mice, increasing DHA to 100 mg can optimize the effect, verifying the necessity of fine-tuning the ratio according to individual genetic background, and having significant cognitive function improvement and neuroprotective effects.

[0155] Since in the above experiments, the capsule drugs were halved according to weight, when actually used, the optimal ranges of the components corresponding to the standard component capsules given in Specific Example 1 are as follows:

[0156] DHA: The optimal amount is 130 - 150 mg, with 130 mg as the standard;

[0157] NMN: The optimal amount is 250 - 300 mg, with 250 mg as the standard;

[0158] Vitamin E: The optimal amount is 5 - 6 mg, with 5 g as the standard;

[0159] Ascidian extract: The optimal amount is 90 - 120 mg, with 90 mg as the standard. Specific Example 4:

[0161] As Figures 1 to 5 shown, based on the content in the above specific examples, the following content is further disclosed:

[0162] Experimental verification was carried out for personalized ratios, and the experimental content is as follows:

[0163] Experimental design: The APOE gene is an important genetic risk factor for Alzheimer's disease. Carriers of the APOE4 allele have a higher risk of getting the disease. Research shows that the brain delivery efficiency of DHA is lower in APOE4 carriers, and higher intake is required to achieve the therapeutic effect. NMN improves nerve function by increasing the level of NAD+, but its direct relationship with the APOE genotype still needs further study.

[0164] Animal model: Wild-type mice and APOE knockout mice (simulating genetic differences) were used, and the grouping is as shown in Table 15 below:

[0165]

[0166] Table 15

[0167] Drug administration: Oral administration was given daily for 3 months;

[0168] Measurement indicators: Behavioral tests: Morris water maze (escape latency), fear conditioning (freezing time), novel object recognition (discrimination index); Biomarkers: Brain Aβ level, tau protein phosphorylation, oxidative stress marker (MDA), inflammatory markers (TNF-α, IL-6), NAD+ level, with special attention paid to the correlation between genotype and efficacy.

[0169] Result analysis:

[0170] The results of the behavioral tests are shown in Table 16 below:

[0171]

[0172] Table 16

[0173] As can be seen from Table 16, wild-type mice performed best under the standard formula, while APOE knockout mice had poorer cognitive function under the standard formula, specifically manifested as longer escape latency, lower freezing time and discrimination index. After personalized adjustment, the cognitive function of APOE knockout mice improved, but did not reach the wild-type level.

[0174] The results of the biochemical markers are shown in Table 17 below:

[0175]

[0176]

[0177] Table 17

[0178] As can be seen from Table 17, the pathological markers (Aβ level, tau phosphorylation, oxidative stress and inflammatory markers) in APOE knockout mice were significantly higher than those in wild-type mice under the standard formula. After personalized adjustment, these markers improved, but the NAD+ level decreased due to the reduction of NMN to 100 mg (150 ± 15 nmol / mg protein), which was lower than that in the standard group (200 ± 20 nmol / mg protein).

[0179] Experimental conclusion: The experimental results show that the standard formula has the best cognitive function and neuroprotective effect on wild-type mice, while APOE knockout mice perform poorly under the standard formula, which is related to the influence of APOE genotype on the transmission and utilization of DHA in the brain. Studies have shown that the brain transmission efficiency of DHA in APOE4 carriers is relatively low. Personalized adjustment (increasing DHA to 100 mg and reducing NMN to 100 mg) significantly improves the cognitive function and pathological markers of APOE knockout mice, but does not reach the wild-type level, and the NAD+ level decreases due to the reduction of NMN. This result supports the necessity of personalized ratio, especially for the adjustment of APOE genotype carriers. Increasing DHA may play a role by improving brain cell membrane function and reducing Aβ deposition, while reducing NMN may be feasible in some cases, but the impact on NAD+ level needs to be weighed.

[0180] The experiment verified the improved efficacy of personalized ratio for APOE knockout mice, especially in terms of cognitive function and pathological markers. However, it should be noted that the reduction of NMN may lead to a decrease in NAD+ level. Future research can further optimize the ratio and explore the best solutions under different genetic backgrounds. Specific Example Five:

[0182] As Figures 1 to 5 shown, based on the content in the above specific examples, the following content is further disclosed:

[0183] The above composition capsules were actually applied. After patients took the composition capsules, various indicators were examined. To further fit human cases, the following cases used the Montreal Cognitive Assessment Scale (MoCA) to score the patients. The specific application cases are as follows:

[0184] The composition capsule consists of ascidian extract, free DHA, NMN, and vitamin E, with a standard capsule content: 90 mg of ascidian extract, 130 mg of free docosahexaenoic acid, 250 mg of β-nicotinamide mononucleotide, and 5 mg of vitamin E. It is prepared by freeze-drying technology. The research further explored the effects of removing ingredients, adjusting doses, personalized ratio, and adding auxiliary ingredients (lecithin, cephalin, ginkgo biloba extract, prebiotics) on the efficacy. The experimental use of APP / PS1 transgenic mouse model verified the irreplaceability and optimal dose range of each ingredient, and personalized adjustment was carried out for APOE genotype.

[0185] Case 1: The standard formula is used for patients with general cognitive decline;

[0186] Patient information: Ms. Li, 65 years old, retired teacher, with recent mild amnesia and inattention, no known genetic variation or allergy history;

[0187] Treatment plan: Take a combination capsule according to the standard formula, containing 90 mg of sea squirt extract, 130 mg of free DHA, 250 mg of NMN, and 5 mg of vitamin E, once a day for 3 months;

[0188] Treatment Testing:

[0189] MoCA score: 24 points in the first month → 26 points in the third month (reaching the normal threshold);

[0190] Biomarkers: Aβ decreased by 38% (260→200pg / mg), tau phosphorylation decreased by 40% (0.5→0.3), and MDA decreased by 33% (3→2nmol / mg);

[0191] Inflammatory markers: TNF-α decreased by 33% (60→40pg / mg), IL-6 decreased by 40% (25→15pg / mg);

[0192] Clinical observations: Orientation (5→6 points) and delayed recall (2→4 points) improved significantly, and calculation and language fluency improved. Ms. Li reported significant improvements in memory and attention, improved scores on job-related tests, and enhanced daily living functions.

[0193] Result analysis: The standard formula is suitable for general patients, and the effect is consistent with the experimental results, which proves the rationality of the initial ingredients of 90 mg of ascidian extract, 130 mg of free docosahexaenoic acid, 250 mg of β-nicotinamide mononucleotide and 5 mg of vitamin E.

[0194] Case 2: Removal of vitamin E for patients allergic to vitamin E;

[0195] Patient information: Mr. Zhang, 70 years old, early-stage Alzheimer's disease patient, allergic to vitamin E, needs to adjust the formula;

[0196] Treatment plan: Adjust the capsule to 90 mg of sea squirt extract, 130 mg of free DHA, and 250 mg of NMN, remove vitamin E, once a day for 3 months;

[0197] Treatment Testing:

[0198] MoCA progress: 18→20 points (still a mild disability);

[0199] Oxidative stress: MDA decreased by only 14% (3.5→3nmol / mg);

[0200] Inflammation rebound: TNF-α maintained at 60pg / mg (no improvement), IL-6 only decreased by 16% (25→21pg / mg);

[0201] Clinical analysis: There was partial improvement in visuospatial / executive function (from 3 to 4 points), but no changes in naming (from 2 to 2 points) and abstraction (from 1 to 1 point). Cognitive function improved, but not as well as with the standard formula. Inflammatory markers were higher, indicating weakened antioxidant protection.

[0202] Result analysis: Removing vitamin E led to a decline in efficacy, consistent with the experimental results, emphasizing the importance of vitamin E in antioxidant and anti-inflammatory effects.

[0203] Case 3: Increasing DHA for patients with higher DHA requirements

[0204] Patient information: Ms. Wang, 55 years old, with low DHA metabolic efficiency, requiring a higher dose to improve cognitive function;

[0205] Treatment plan: The capsule was adjusted to 130 mg of ascidian extract, 150 mg of free DHA, 250 mg of NMN, and 5 mg of vitamin E, once a day for 3 months;

[0206] Treatment test:

[0207] MoCA dynamics: 22 → 28 points (reaching excellent cognitive level);

[0208] Neuroregeneration index: BDNF increased by 120% (from 12 to 26.4 pg / ml);

[0209] DHA bioavailability: The proportion of DHA in the erythrocyte membrane increased from 3.8% to 5.2%;

[0210] Clinical characteristics: Attention (from 5 to 6 points) and delayed recall (from 3 to 5 points) improved significantly. MRI showed a 2.8% increase in hippocampal volume. Cognitive function improved significantly, and brain imaging showed enhanced activity in memory and learning-related regions, better than the standard dose;

[0211] Result analysis: Increasing DHA to 150 mg was consistent with the dose optimization experimental results and was particularly suitable for patients with low DHA metabolic efficiency.

[0212] Case 4: Individualized formulation for APOE4 genotype carriers;

[0213] Patient information: Mr. Chen, 60 years old, APOE4 genotype carrier, with a family history of Alzheimer's disease;

[0214] Treatment plan: Individualized capsule containing 130 mg of ascidian extract, 150 mg of free DHA, 300 mg of NMN, and 6 mg of vitamin E, once a day for 6 months;

[0215] Treatment test:

[0216] MoCA progress: 19 → 25 points (close to the normal lower limit);

[0217] Gut-brain axis improvement: Fecal SCFA increased 3-fold and LPS-binding protein decreased by 45%;

[0218] Gene-specific response: Activity of the APOE4-related inflammatory pathway (NF-κB) decreased by 60%;

[0219] Clinical observation: Verbal fluency (2 → 4 points) and abstract thinking (1 → 3 points) were significantly improved; Cognitive function tests showed significant improvement, and biomarker levels were close to the standard treatment effect of non-APOE4 patients;

[0220] Result analysis: Personalized adjustment was consistent with the experimental results, and increasing DHA and adjusting NMN optimized the efficacy of APOE4 carriers.

[0221] Case 5: Controlled-release formulation for patients with poor medication compliance;

[0222] Patient information: Ms. Liu, 65 years old, moderate Alzheimer's disease, difficult to take medicine on time, need to improve medication compliance;

[0223] Treatment plan: Use controlled-release formulation capsules containing standard ingredients (90 mg of ascidian extract, 130 mg of free docosahexaenoic acid, 250 mg of β-nicotinamide mononucleotide, and 5 mg of vitamin E), release time of 12 hours, once a day for 3 months;

[0224] Treatment test:

[0225] The fluctuation index (DF) of DHA blood drug concentration decreased from 85% to 35%;

[0226] The peak time of NMN was extended from 1.2 h to 6.8 h;

[0227] Clinical effect: MoCA score: 16 → 22 points (3 months); Medication compliance: Increased to 92%; Circadian rhythm improvement: The 24-hour salivary cortisol curve returned to the normal rhythm, blood drug concentration was stable, cognitive function remained consistent, family members reported less confusion, and the disease management effect was better;

[0228] Result analysis: The controlled-release formulation is suitable for patients with poor medication compliance, provides continuous efficacy, and meets the expected effect of microcapsule technology.

[0229]

[0230] Table 18

[0231] Table 18 above shows the scoring dimensions and judgment criteria of the Montreal Cognitive Assessment Scale (MoCA). Through multi-dimensional MoCA assessment, it can more accurately reflect the action mode of the composition capsule on specific cognitive domains, providing a quantitative basis for clinical personalized adjustment.

[0232] The above five cases cover different scenarios of the standard application, ingredient adjustment, personalized ratio, and controlled-release formulation of the composition capsule. The research shows that the standard formulation is suitable for general patients, removing ingredients may reduce the efficacy, personalized adjustment (APOE4 genotype) can optimize the cognitive enhancement effect in specific populations, and the controlled-release formulation improves medication compliance, verifying the versatility and pertinence of the composition capsule, which is applicable to the prevention and early treatment of Alzheimer's disease.

[0233] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or further includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "including a reference structure" does not exclude the existence of additional identical elements in the process, method, article or device comprising the element.

[0234] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a composition capsule containing ascidian extract, DHA and NMN, characterized in that: The composition capsule is composed of sea squirt extract, free docosahexaenoic acid, nicotinamide mononucleotide and vitamin E. The preparation method of the composition capsule is as follows: Sp1: Sea squirt extract: Using sea squirt as the extraction raw material, a mixed solvent of alcohol entrainer and supercritical CO2 is used to remove the grease and neutral fat in the sea squirt. A mixed solvent of alcohol entrainer and supercritical CO2 is used to extract the phospholipid active substances with relatively large polarity to obtain the sea squirt extract. The extraction environment is 40 °C and 300 bar; Sp2: Preparation of the content: Mix the sea squirt extract, free DHA, NMN and vitamin E to obtain a preliminary mixed content. Then, use a colloid mill to perform three homogenization treatments on the preliminary mixed content. Then, place the homogenized preliminary mixed content in a stirring tank and stir for 40 - 60 min, and then perform filtration treatment to obtain a content with uniform components for standby; Sp3: Sol preparation: Add glycerol and purified water to a gelatinizing tank, heat to 70 - 80 °C, then add gelatin, stir while adding, and perform heat preservation treatment until all the gelatin is dissolved to obtain a mixed gelatin solution; Sp4: Vacuum degassing: Place the mixed gelatin solution under vacuum conditions until no bubbles are generated on the surface of the mixed gelatin solution. After adjusting the viscosity of the mixed gelatin solution, place the filtered mixed gelatin solution in a heat preservation bucket and let it stand at 65 °C ± 8 °C for no less than 2 h to obtain the capsule material; Sp5: After the capsule material obtained in step Sp4 and the content obtained in step Sp2 are subjected to pill pressing, shaping, drying, pill selection and packaging, fish oil soft capsules are obtained.

2. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: The component compatibility of sea squirt extract, free docosahexaenoic acid, nicotinamide mononucleotide and vitamin E in the composition capsule is: 14 - 20 parts of sea squirt extract, 20 - 40 parts of free docosahexaenoic acid, 40 - 53 parts of nicotinamide mononucleotide and 0.2 - 2 parts of vitamin E, and the total component compatibility of the composition capsule is 100 parts.

3. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: The extraction time for extracting the sea squirt extract is 2 - 4 h, the CO2 flow rate is 10 - 15 L / min, 5% - 10% ethanol is added as a co-solvent during the extraction process, the content of plasmalogen in the obtained sea squirt extract is not less than 1%, the extraction pressure is 10 - 45 MPa, and the extraction temperature is 30 - 50 °C.

4. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: The raw material of the free docosahexaenoic acid is fish oil, which is refined by supercritical fluid extraction technology to remove heavy metals and impurities.

5. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: Lecithin and cephalin are further added to the composition capsule as auxiliary phospholipid components, where the content of lecithin is 10% - 20% of the sea squirt extract, and the content of cephalin is 5% - 10% of the sea squirt extract.

6. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: Ginkgo biloba extract is added to the composition capsule as an additional neuroprotective component, and the content of ginkgo biloba extract is 5% - 10% of the total weight of the composition.

7. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: Prebiotics are added to the composition capsule as an enteric-brain axis regulating component, and the content of prebiotics is 10% - 15% of the total weight of the composition.

8. A method for preparing a composition capsule containing ascidian extract, DHA and NMN according to claim 1, characterized in that: The ratio range of the sea squirt extract to the free docosahexaenoic acid in the combined capsule is between 1:3 and 1:1.

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