A composition for improving mild cognitive impairment

Abstract

The present invention relates to a composition containing krill oil, folic acid and vitamin D. The advantages of the present invention are as follows: The composition can protect cardiovascular health. While meeting the needs of supplementing folic acid and vitamin D, it can synergistically improve the symptoms of mild cognitive impairment, enhance memory, and reduce the risk of dementia. In addition, by optimizing the proportion of components in the composition and reducing the dose of krill oil, the risk of arsenic element accumulation in the body that may be caused by long-term high-dose intake of krill oil can be reduced, as well as the risk of cognitive impairment and sports injuries caused by excessive arsenic.

Description

Technical Field

[0001] The present invention relates to a composition containing krill oil, folic acid and vitamin D and its use, belonging to the technical field of great health. Background Art

[0002] With the extension of the population lifespan and the aging of society, the prevalence of Alzheimer's disease (AD) is on the rise globally. Although countries around the world have invested a large amount of manpower and material resources in the research and development of AD treatment drugs, no drug that can stop the progression of AD has been developed yet. Therefore, AD prevention has gradually shifted to the pre-AD stage - mild cognitive impairment (MCI). MCI is an intermediate stage from normal cognition to dementia onset. Patients have mild memory impairment, but their daily living abilities are basically retained and do not meet the dementia diagnosis criteria. As the condition worsens, patients gradually show further decline in cognition and daily living abilities, rely on others for care, and transform into dementia. Currently, it is generally agreed that MCI is an important transitional stage in the pre-dementia stage of AD, and MCI patients are at high risk of developing AD. Preventing and intervening in MCI is of great significance for delaying the onset of dementia and improving the quality of life.

[0003] Krill oil is an extract extracted from Antarctic krill species and contains ω-3 essential fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). These polyunsaturated fatty acids have the function of regulating blood lipids, are beneficial to promoting human health, and have certain protective effects on the cardiovascular system, nervous system, bones and joints, vision, skin, etc. However, krill contains relatively more arsenic, mainly because arsenic elements existing in sedimentary rocks and pyroclastic rocks in nature enter seawater mainly through surface runoff and other ways, and are enriched in marine organisms through the food chain. At the same time, most arsenic compounds in Antarctic krill will migrate to krill oil during the extraction process. Folic acid belongs to B vitamins and is an essential substance for the growth and reproduction of body cells. Lack of folic acid in pregnant women may lead to fetal neural tube malformation, megaloblastic anemia, fetal growth retardation, etc. Vitamin D is a fat-soluble vitamin, which has the functions of promoting calcium and phosphorus absorption, bone matrix calcification and beneficial bone formation, can promote bone development, and prevent rickets in children and osteoporosis in middle-aged and elderly people. Summary of the Invention

[0004] We unexpectedly found in scientific experiments that when krill oil, folic acid, and vitamin D are used in combination, they can synergistically improve the symptoms of movement, behavior, and cognitive impairment in elderly MCI rats. By optimizing the composition ratio of the composition and reducing the dosage of krill oil, on the one hand, the cost is reduced, and on the other hand, through the combination with folic acid and vitamin D, it is used for the supplementation of folic acid and vitamin D in the MCI population, improving the phenomenon of single nutritional components in current krill oil products, better assisting in the prevention and improvement of cognitive dysfunction, and achieving the long-term goal of delaying the onset of dementia, which is a significant improvement over existing krill oil products. In addition, it was found that the combination of high-dose krill oil with folic acid and vitamin D can significantly promote the metabolic excretion of arsenic in krill oil, reduce the risk of arsenic accumulation in the body caused by long-term and large-dose intake of krill oil, and improve the food safety.

[0005] The present invention adopts the following technical solutions:

[0006] A composition for improving mild cognitive impairment, wherein the composition consists of krill oil: folic acid substance: vitamin D in a weight ratio of: 200 - 1000: 0.08 - 0.5: 0.002 - 0.02.

[0007] In the composition of the present invention, as a preference, the weight ratio of krill oil: folic acid substance: vitamin D is: 500 - 1000: 0.1 - 0.4: 0.002 - 0.01.

[0008] In the present invention, the "parts" can be micrograms, milligrams, grams, microliters, milliliters, deciliters, etc.

[0009] In the composition of the present invention, the krill oil is selected from one or more of Antarctic krill oil, deep-sea krill oil, and krill oil containing ω-3.

[0010] In the composition of the present invention, the folic acid substance is selected from one or more of 5-methyltetrahydrofolic acid, calcium formyltetrahydrofolate, dihydrofolic acid, folic acid, or the active metabolite of folate salt and the substance that can release / generate folic acid substance in the body.

[0011] In the composition of the present invention, the vitamin D is selected from vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), or various forms of vitamin D from other sources.

[0012] In the composition of the present invention, the krill oil contains DHA, and the DHA accounts for more than 5% of the weight of the krill oil.

[0013] The composition described in the present invention may also contain acceptable excipients or carriers or mixtures thereof, and the acceptable excipients or carriers or mixtures thereof include one or more of food sugars or functional sweeteners, fillers, wetting agents, binders, and lubricants.

[0014] The dosage forms of the composition described in the present invention are oral preparations such as tablets, capsules, gummies, liquid preparations, granules, syrups, and powders.

[0015] In the present invention, the composition may exist in the form of drugs and the like.

[0016] Use of the composition described in the present invention in the preparation of a product for preventing or improving symptoms of mild cognitive impairment of the brain.

[0017] Improving brain cognitive function in the use of the composition described in the present invention includes improving symptoms such as cognitive function decline caused by anxiety disorders, depression, schizophrenia, autism, juvenile hyperactivity disorder, inability to concentrate, Alzheimer's disease (AD), Parkinson's disease, multiple sclerosis (MS), postoperative cognitive dysfunction (POCD), spinal cord injury (SCI), AIDS dementia complex (ADC), ischemia, stroke, traumatic brain injury (TBI), brain or central nervous system infections, and other diseases.

[0018] Compared with the prior art, the present invention provides a composition containing krill oil, and the components produce a significant synergistic effect in improving brain cognitive function, and long-term or high-dose application will not cause arsenic element accumulation, which is safer and more effective, and is a significant improvement over the prior art. Detailed implementation manners

[0019] The following examples only represent the preferred implementation manners of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the invention patent shall be subject to the appended claims.

[0020] The raw materials used in the composition, use, and product provided by the present invention can all be purchased from the market. The folic acid involved in the examples was purchased from DSM; the krill oil was purchased from Xi'an Weite Biotechnology Co., Ltd.; the vitamin D was purchased from DSM.

[0021] Example 1: Effects of the composition of krill oil, folic acid, and vitamin D on rats with mild cognitive impairment in old age

[0022] We designed the Morris water maze test to detect the effect of the composition of the present invention on the cognitive function of rats through behavioral experiments. At the same time, the contents of malondialdehyde (MDA) and glutathione (GSH) in the cerebral cortex and hippocampus of rats were detected by colorimetry to evaluate the oxidative stress function of the brain tissue.

[0023] Experimental animals and grouping: SPF-grade Wistar rats, half male and half female, 18 - 20 months old, were raised in an environment with a room temperature of 18 - 28 °C and a relative humidity of 40% - 70%, with free access to food and water. After 1 week of adaptive feeding with normal feed, 10 rats were randomly selected as the blank control group, and the rest were modeled according to the following method.

[0024] Preparation of animal model: Refer to the method of incomplete occlusion of the common carotid artery in rats with controllable stenosis degree for modeling. Fast for 8 - 12 h before surgery, allow free drinking water, anesthetize by intraperitoneal injection of 10% chloral hydrate (3 ml / kg), routinely prepare the skin in the middle of the neck for disinfection and incision, with an incision of 2 - 3 cm, bluntly separate the bilateral common carotid arteries, and parallelly place a No. 0.45 injection needle (causing severe stenosis of the common carotid artery) at about 1.5 cm from the bifurcation of the internal carotid artery and external carotid artery on the proximal side of the common carotid artery for the model control group and the experimental group rats, gently tie the common carotid artery and the syringe needle with 6-0 silk thread, then carefully pull out the needle, and the blood flow resumes while the coil is still ligated. Intermittently suture the skin and routinely disinfect the wound. In the blank control group, only the bilateral common carotid arteries were separated but not ligated. Intramuscular injection of penicillin was given continuously for 5 days after surgery to prevent infection. After 30 days of modeling, the Morris water maze test was performed to detect the behavioral indexes of rats in each group. Taking the average value of the escape time of rats in the blank control group as the reference value, calculate the ratio of the difference between the average escape latency of each rat in the remaining groups at each time period and the reference value to the average escape latency time of the rat. A value < 20% was defined as rats with mild cognitive impairment.

[0025] Behavioral detection: After the Morris water maze test confirmed the success of modeling, random grouping and drug administration were carried out according to Table 1 below, and rats in each group were continuously gavaged for 30 days. After 30 days of gavage, the Morris water maze experiment was performed on rats in each group to detect learning and memory abilities. The experiment lasted for 5 days. On the first day, rats were allowed to swim freely for 2 min to familiarize themselves with the environment, and the formal experiment started on the second day. Experimental content: ① Place navigation experiment: Used to detect the spatial learning ability of rats. If the rat did not find the platform within 90 s, it was guided to the platform and allowed to stay on the platform for 10 s, and the latency at this time was recorded as 90 s, and the average search latency of the platform was observed. ② Spatial exploration experiment: Used to detect the spatial memory ability of rats. After removing the platform, the swimming trajectory of the rat within 60 s was recorded, and the number of times the rat crossed the original platform position and the percentage of swimming time in the original platform quadrant were used as observation indexes.

[0026] Detection of malondialdehyde and glutathione: After the Morris water maze experiment, the animals were sacrificed, and the brain was quickly removed and rinsed with cold saline to remove blood and impurities. The hippocampus and cortical tissues were separated, weighed, and made into 10% tissue homogenates with normal saline at 4°C. The tissue homogenates were centrifuged at 4000 g for 10 min, and the supernatants were taken for detection. The detection of malondialdehyde (MDA) and glutathione (GSH) was carried out strictly according to the instructions of the kit.

[0027] Experimental results: To confirm the scientificity of the pharmaceutical composition provided by the present invention, and to illustrate that the three components of the pharmaceutical composition are rationally formulated and can exert a synergistic effect when combined with each other, rather than a simple superposition of pharmacological effects, the Jin Zhengjun Q value method was introduced for analysis. The Jin Zhengjun Q value method is also known as the probability addition method. According to the pharmacological effects of the combination of two drugs and the pharmacological effects of the two drugs used alone in the dose-effect curve region, it is calculated using the following formula: Q = E A+B / (E A +E B -E A *E B ), where the numerator represents the "measured combined effect" and the denominator represents the "expected combined effect". (To satisfy the analysis of the pharmacological effect relationship between the components and the composition, their pharmacological effects are transformed into effects that can intuitively reflect the strength of the pharmacological effect. The calculation formula is: E i = 1 - P i / P 模型组 , P i is the pharmacological index of each component, and P 模型组 is the pharmacological index of the model group), and Q is the ratio of the two: When Q is less than 0.85, the combination of the two drugs is considered an antagonistic effect; when it is less than 1.15 and greater than 0.85, it is considered an additive effect; when it is greater than 1.15, it is considered a synergistic effect. Based on the calculation formula for the combination of two drugs, the pharmacological effects of the combination of three drugs and the pharmacological effects of the three drugs used alone are calculated using the following formula: Q = E A+B+C / (E A +E B +E C -E A *E B -E A *E C -E B *E C -E A *E B *E C ),

[0028] As shown in the experimental results in Table 1. In the place navigation experiment, after appropriate training, rats can gradually form spatial memory, and their ability to search for the target gradually increases. The average latency of each group of rats to search for the platform gradually shortens. Compared with the blank control group, the average escape latency of the rats in the model control group in the water maze place navigation experiment significantly increases, indicating that the model of rats with mild cognitive impairment in old age is successfully established.

[0029] When Antarctic krill oil was used alone, it showed a significant effect of reducing the escape latency on the 4th and 5th days. Folic acid (FA) or vitamin D (V D ) had no significant effect at the administered doses. By calculating the Q value for the pairwise combination of drugs, except for the Q value of 1.18 for krill oil + folic acid in the test on the 5th day, no synergistic effect was observed in the data of other groups (Q < 1.15). As can be seen from Table 1, for the effect of the triple-drug combination on the average escape latency, through the Q value analysis of the effects of the three single drugs, on the 3rd - 5th days, the Q values were all greater than 1.15, showing an obvious synergistic effect.

[0030] Under the dose conditions of 20 - 100 mg / kg of krill oil + 0.008 - 0.04 mg / kg of folic acid + 0.0002 - 0.002 mg / kg of vitamin D, the average escape latency of rats significantly decreased. Increasing the dose did not produce a further effect, and too low a dose did not produce a significant effect.

[0031] As shown in Table 2 for the results of the effects on the number of times of crossing the original platform and the percentage of swimming time in the original platform quadrant in the water maze spatial exploration experiment, a similar conclusion was obtained as in Table 1 above: The combination of Antarctic krill oil, folic acid, and vitamin has an obvious synergistic effect in increasing the number of times of crossing the original platform and the percentage of swimming time in the original platform quadrant in the water maze spatial exploration experiment of rats with mild cognitive impairment in old age, indicating that the composition of this invention has an obvious effect in improving mild brain cognitive impairment.

[0032] Table 1 Average escape latency of rats in each group in the water maze place navigation experiment( n = 9 - 10)

[0033]

[0034] Note: Compared with the blank control group, bb P < 0.01; compared with the model group, a P < 0.05, aa P < 0.01. The Q value is calculated for the effect of the composition and the single-component group.

[0035] Table 2 Situation of rats in each group in the water maze spatial exploration experiment( n = 9 - 10)

[0036]

[0037] Note: Compared with the blank control group, bb P < 0.01; compared with the model group, a P < 0.05, aa P < 0.01. The Q value is calculated for the effect of the composition and the single - component group.

[0038] Example 2: Determination of Arsenic Content in the Composition of Krill Oil, Folic Acid and Vitamin D

[0039] To illustrate that the krill oil, folic acid and vitamin D composition of the present invention can reduce the risk of a large accumulation of arsenic in the body caused by long - term consumption of krill oil, we designed an experiment to continuously intragastrically administer for 28 days to determine the arsenic content.

[0040] 28 - day continuous intragastric administration experiment: Twenty healthy adult Wistar rats, with 10 males and 10 females, were selected. Thirty healthy Wistar rats were randomly divided into 3 groups, namely the blank control group, the Antarctic krill oil group, and the Antarctic krill oil + folic acid + vitamin D group, with 10 rats in each group. 12 hours before the experiment, the rats were fasted but allowed to drink water. They were administered drugs according to the doses in Table 3 below. The rats in each group were continuously intragastrically administered for 4 weeks. 24 hours after the last intragastric administration, the rats were anesthetized with sodium pentobarbital, and blood was taken from the abdominal aorta to sacrifice the rats. The liver and kidneys were dissected and stored at - 80 °C for later use.

[0041] Determination of arsenic content: Weigh 0.1 g of each of the liver and kidney, add 1 ml of normal saline to make a homogenate. Take 0.2 ml of the homogenate from each organ and add 1 ml of a mixed acid of nitric acid: perchloric acid (volume ratio 3:1); take 0.5 ml of heparin - anticoagulated whole blood and 1 ml of the mixed acid; take about 1 ml of urine sample, add 0.25 ml of nitric acid and 0.5 ml of hydrogen peroxide solution, and heat and digest on an electric hot plate until colorless crystals are formed. After dissolving with deionized water, transfer it into a colorimetric tube, and use hydride generation - atomic fluorescence spectrometry to determine the arsenic content in each organ tissue.

[0042] Table 3 Arsenic content in the liver and kidney of rats fed for 28 days ( n = 10)

[0043]

[0044] Note: Compared with the Antarctic krill oil group, aa P < 0.01.

[0045] In this example, considering that the liver is the most important metabolic organ in animals and humans, and arsenic metabolism mainly occurs in the liver, while the kidney is the main excretory organ of arsenic, the arsenic contents in the livers and kidneys of rats were measured. As shown in the experimental results of Table 3. After continuous intragastric administration for 28 days, there were significant differences in the arsenic contents in the livers and kidneys of rats in the Antarctic krill oil group compared with those in the control group (P≤0.01), indicating that long-term administration of a large dose of Antarctic krill oil resulted in the accumulation of arsenic in the liver and kidney tissues of rats. There were significant differences in the arsenic contents in the liver and kidney tissues between the Antarctic krill oil + folic acid + vitamin D group and the Antarctic krill oil group (P≤0.01). It can be seen that the Antarctic krill oil + folic acid + vitamin D group can significantly reduce the arsenic contents in the liver and kidney tissues of rats when excessive krill oil is used. The Antarctic krill oil + folic acid + vitamin D composition has the effect of reducing the arsenic contents in the livers and kidneys of rats, which may be related to the fact that folic acid is one of the main sources of methyl donors in food, can affect arsenic methylation metabolism, reduce blood arsenic content, promote arsenic excretion, and reduce body accumulation.

Claims

1. A composition for improving mild cognitive impairment of the brain, which is composed of krill oil, folic acid substances and vitamin D. The weight ratio of krill oil: folic acid substances: vitamin D is 200 - 1000: 0.08 - 0.4: 0.002 - 0.02, wherein the krill oil is Antarctic krill oil and the folic acid substances are folic acid.

2. The composition according to claim 1, wherein The weight ratio of krill oil: folic acid substances: vitamin D is 500 - 1000: 0.1 - 0.4: 0.002 - 0.

01.

3. The composition according to claim 1 or 2, characterized in that: The krill oil contains DHA, and the DHA accounts for more than 5% of the weight of the krill oil.

4. The composition according to claim 1, characterized in that, The dosage form of the composition is one of tablets, capsules, gummies, liquid preparations, granules, syrups and powders.

5. Use of the composition according to any one of claims 1 - 4 in the preparation of a product for improving mild cognitive impairment of the brain.

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