Application of caffeoyl butanediamine in preparation of drugs, health care products or functional food for preventing and treating Alzheimer's disease

Abstract

The invention discloses application of caffeoyl butanediamine in preparation of drugs, health care products or functional food for preventing and treating Alzheimer's disease. Belongs to the technical field of biological medicine. Animal experiments prove that caffeoyl butanediamine can prevent and treat Alzheimer's disease, can significantly improve the working memory, spatial memory and learning ability of mice with Alzheimer's disease (AD) induced by combination of D-galactose and aluminum trichloride, and can significantly reduce the levels of Abeta40 and Abeta42 of brain tissues of the mice with AD. The caffeoyl butanediamine provided by the invention can be used for preventing and treating neurodegenerative diseases such as Alzheimer's disease.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and more specifically relates to the application of caffeoylbutanediamide in the preparation of drugs, health products or functional foods for preventing and treating Alzheimer's disease. Background Art

[0002] Alzheimer's disease (AD), also known as senile dementia, is a type of disease characterized by progressive cognitive impairment and memory loss, accompanied by neuronal dysfunction and even neuronal death. Currently, there are approximately 36.5 million AD patients globally, and the number of patients in China alone accounts for one-sixth of the total. With the aggravation of population aging, the number of AD patients will increase exponentially. It is estimated that the number of global patients will exceed 100 million by 2050. As the country with the largest total number of AD patients in the world, China will have more than 20 million AD patients, and the incidence rate among the elderly over 85 years old will exceed 1 / 3. It has become one of the important diseases threatening the health of the elderly, second only to heart disease, cancer and stroke (stroke), seriously affecting the quality of life of the elderly, and bringing a heavy medical burden and serious social problems. Thus, the prevention and treatment of AD are urgent problems to be solved in the medical field.

[0003] The pathogenesis of AD is complex, and its core pathological basis mainly involves the following three categories: one is the neurofibrillary tangles in the brain; the second is the abnormal aggregation of tau protein in neuronal cells in the brain; the third is the accumulation of β-amyloid peptide (Aβ) in the brain and hippocampal region, and the formation of senile plaques outside the cells. Due to the complexity of its etiology, there is no treatment plan in the existing treatment options that can completely cure AD, and most of them only control the aggravation of the condition.

[0004] The existing drugs for treating AD are mainly cholinesterase inhibitors and N-methyl-D-aspartic acid (NMDA) receptor antagonists, such as tacrine, donepezil, memantine, etc., which can temporarily relieve symptoms but are accompanied by serious side effects. It is worth noting that in recent years, significant breakthroughs have been made in the field of natural medicine research. A large number of experimental studies have shown that a variety of active monomer components of traditional Chinese medicine can show significant efficacy in the prevention and treatment of AD by inhibiting the activity of cholinesterase and other mechanisms. These important findings not only open up new research directions for the prevention and treatment of AD, but also make the screening and development of monomer components with anti-AD activity from the treasure house of natural medicine become the current forefront hot spot in drug research.

[0005] Caffeoylbutanediamide is a monomer active ingredient isolated by the inventor from traditional Chinese medicines such as Cortex Lycii Radicis and Fructus Lycii. Currently, there are few research reports on the biological activity of caffeoylbutanediamide, and there is no relevant record of caffeoylbutanediamide being used for the prevention and treatment of AD. Summary of the Invention

[0006] In view of this, the present invention provides the application of caffeoylbutanediamide in the preparation of medicaments, health products or functional foods for preventing and treating Alzheimer's disease. Through previous animal experiments, the present invention found that caffeoylbutanediamide has a significant effect on alleviating Alzheimer's disease. Further, developing it into a drug that can be used to prevent and treat Alzheimer's disease, or a health product or functional food that can prevent and treat Alzheimer's disease, will have great clinical application value.

[0007] Caffeoylbutanediamide, with the molecular formula C 13 H 18 N2O3, the English name is N-(4-aminobutyl)-3-(3,4-dihydroxyphenyl)propenamide, the English abbreviation is N-caffeoylputrescine or Caffeoylputrescine, the molecular weight is 250.14, and its chemical structural formula is shown in Formula (1):

[0008] Formula (1) In order to achieve the above object, the present invention adopts the following technical solutions: The application of caffeoylbutanediamide in the preparation of medicaments for preventing and treating Alzheimer's disease, or in the preparation of health products or functional foods for preventing and treating Alzheimer's disease.

[0009] Further, caffeoylbutanediamide can significantly increase the spontaneous alternation response rate of Alzheimer's disease mice in the Y-maze experiment, that is, improve their working memory ability.

[0010] Further, caffeoylbutanediamide can significantly increase the ability of Alzheimer's disease mice to recognize new things, that is, improve their learning ability.

[0011] Further, caffeoylbutanediamide can significantly reduce the escape latency of Alzheimer's disease mice in the Morris water maze experiment and increase the number of times of crossing the platform, that is, improve their spatial memory ability.

[0012] Further, caffeoylbutanediamide can significantly reduce the content of Aβ 40 and Aβ 42 in the brain tissue of Alzheimer's disease mice.

[0013] A medicament for preventing and treating Alzheimer's disease, characterized in that it comprises caffeoylbutanediamide.

[0014] Further, the dosage form is an orally administrable dosage form, an external patch or an injection dosage form permitted in pharmacy.

[0015] A health care product for preventing and treating Alzheimer's disease, comprising cafebutanediamide.

[0016] A functional food for preventing and treating Alzheimer's disease, comprising cafebutanediamide.

[0017] Through the above technical solutions, compared with the prior art, the beneficial effects obtained by the present invention are as follows: in the dose range of 10 - 25 mg / kg given to experimental animals, cafebutanediamide can significantly improve the working memory, spatial memory and learning ability of D-galactose combined with aluminum trichloride-induced AD mice, and significantly reduce Aβ in the brain tissue of AD mice 40 and Aβ 42 levels. The above all confirm that cafebutanediamide has the effect of preventing and treating AD and can be used for the prevention and treatment of neurodegenerative diseases such as AD. Brief Description of the Drawings

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained according to the provided drawings.

[0019] Figure 1 Effect of cafebutanediamide on the body weight of AD mice (C is the normal group, M is the model group, P is the positive drug donepezil group, NCP-H is the high-dose cafebutanediamide group, NCP-L is the low-dose cafebutanediamide group; compared with the normal group, ## p <0.01; compared with the model group, ** p <0.01) Figure 2 Effect of cafebutanediamide on the spontaneous alternation response rate of AD mice investigated by the Y-maze experiment (C is the normal group, M is the model group, P is the positive drug donepezil group, NCP-H is the high-dose cafebutanediamide group, NCP-L is the low-dose cafebutanediamide group; compared with the normal group, ## p <0.01; compared with the model group, ** p <0.01, *** p <0.001) Figure 3 Effect of cafebutanediamide on the new object discrimination ability of AD mice investigated by the novel object recognition experiment (C is the normal group, M is the model group, P is the positive drug donepezil group, NCP-H is the high-dose cafebutanediamide group, NCP-L is the low-dose cafebutanediamide group; compared with the normal group, #p < 0.05; compared with the model group, * p < 0.05, *** p < 0.001) Figure 4 To investigate the effects of cafoylputrescine on the escape latency and the number of platform crossings of AD mice in the Morris water maze experiment (C is the normal group, M is the model group, P is the positive drug donepezil group, NCP-H is the high-dose cafoylputrescine group, NCP-L is the low-dose cafoylputrescine group; compared with the model group, * p < 0.05, ** p < 0.01) Figure 5 To investigate the effects of cafoylputrescine on Aβ in the brain tissue of AD mice 40 and Aβ 42 content (C is the normal group, M is the model group, P is the positive drug donepezil group, NCP-H is the high-dose cafoylputrescine group, NCP-L is the low-dose cafoylputrescine group; compared with the normal group, ### p < 0.001; compared with the model group, ** p < 0.01, *** p < 0.001) Detailed implementation methods

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to 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 of 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.

[0021] The cafoylputrescine used in the following examples is the compound shown in the above formula (1), which can be obtained by commercial purchase or experimental self-preparation.

[0022] The medicaments required for the present invention are conventional experimental medicaments, which are purchased from commercial channels; the experimental methods not mentioned are conventional experimental methods and will not be elaborated here one by one. Example 1

[0023] A drug for preventing and treating Alzheimer's disease, in an oral dosage form, comprising cafoylputrescine, wherein the single application dose of cafoylputrescine is 10 mg / kg. Example 2

[0024] A drug for preventing and treating Alzheimer's disease, in an oral dosage form, comprising cafoylputrescine, wherein the single application dose of cafoylputrescine is 25 mg / kg. Example 3

[0025] A health care product for preventing and treating Alzheimer's disease, in the form of an oral health care product, comprising cafebutyramide, wherein the single application dose of cafebutyramide is 25 mg / kg. Example 4

[0026] A functional food for preventing and treating Alzheimer's disease, in oral form, comprising cafebutyramide, wherein the single application dose of cafebutyramide is 10 mg / kg. Example 5

[0027] A medicine for preventing and treating Alzheimer's disease, in injection form, comprising cafebutyramide, wherein the single application dose of cafebutyramide is 10 mg / kg. Example 6

[0028] A medicine for preventing and treating Alzheimer's disease, in the form of a topical patch, comprising cafebutyramide, wherein the single application dose of cafebutyramide is 25 mg / kg.

[0029] The following animal experiments further illustrate the effects of the above Examples 1 to 6: I. Animal experiment design A mouse model of Alzheimer's disease (AD) was induced by D-galactose combined with aluminum trichloride (AlCl3). After intragastric administration of cafebutyramide, the preventive and therapeutic effects of cafebutyramide on Alzheimer's disease were evaluated by mouse body weight, Y-maze experiment, novel object recognition experiment, Morris water maze experiment, Aβ 40 and Aβ 42 content.

[0030] II. Experimental procedure 1. Experimental animals Healthy SPF-grade ICR mice, male, weighing 20 - 30 g, were provided by the Experimental Animal Center of Ningxia Medical University (Experimental Animal Use License SYSK (Ning) 2020 - 0001), and were housed in an SPF-grade barrier environment with 12 h of light and 12 h of darkness, and free access to food and water.

[0031] 2. Experimental method The D-galactose combined with AlCl3 was used to induce an Alzheimer's disease (AD) mouse model. After 48 mice were adaptively fed for 3 days, they were randomly divided into 5 groups according to their body weights: a normal group (saline), a model group (saline), a donepezil positive drug group (5 mg / kg), a low-dose cafebutyramide group (NCP-L, 10 mg / kg / d), and a high-dose cafebutyramide group (NCP-H, 25 mg / kg / d). Donepezil and cafebutyramide were both dissolved in saline and administered by gavage at a dose of 10 ml / kg. The mice were subcutaneously injected with D-galactose (200 mg / kg) at a fixed time point every day, and at the same time, 150 mg / kg AlCl3 was administered by gavage to induce the AD model (after 5 weeks of modeling, the success of the modeling was judged by behavioral experiments). The mice in the normal group were subcutaneously injected and gavaged with an equal volume of saline; 4 hours later, the drugs were administered by gavage. The above modeling and gavage were both performed once a day for 10 consecutive weeks of modeling and drug administration.

[0032] 3. Y-maze experiment After 10 weeks of drug administration to each group of mice, the Y-maze experiment was used to investigate the effect of cafebutyramide on the working memory ability of AD mice, and the spontaneous alternation response rate of each mouse was calculated for evaluation.

[0033] 4. Novel object recognition experiment After 10 weeks of drug administration to each group of mice, the novel object recognition experiment was used to investigate the effect of cafebutyramide on the learning and memory ability of AD mice, and the preference index of each mouse was calculated for evaluation.

[0034] 5. Morris water maze experiment After 10 weeks of drug administration to each group of mice, the Morris experiment (continuously measured for 5 days) was used to investigate the effect of cafebutyramide on the spatial memory ability of AD mice, and the escape latency and the number of times of crossing the platform of each mouse were used for evaluation.

[0035] 6. Aβ 40 and Aβ 42 Content determination After 10 weeks of drug administration to each group of mice and completion of the behavioral tests, serum was collected, and then the mice were decapitated under the neck and sacrificed. The whole brain was taken immediately on ice and frozen in liquid nitrogen, and then transferred to -80 °C for storage and standby after the experiment. The mouse brain tissue was ground evenly with a tissue grinder to make a brain tissue homogenate, centrifuged at 5000 xg / min for 10 min at 4 °C, the supernatant was collected, and a kit was used to detect the content of Aβ 40 and Aβ 42 in the supernatant.

[0036] 7. Data processing For data processing of the present invention, ImageJ and Graphpad Prism 9.5 were used for data analysis and statistics. The obtained results were all expressed as mean ± standard deviation (X±S). One-way ANOVA was used for data processing of inter-group difference comparison, and p<0.05 was used as the standard for statistical significance of differences.

[0037] III. Experimental Results 1. Effect on the body weight of mice The results of the effect of cafoyl succinamide on the body weight of mice are shown in Figure 1 as follows.

[0038] After 10 weeks of drug administration to each group of mice, compared with the control group, the body weight of the mice in the model group was significantly reduced ( p <0.01). Compared with the model group, the body weight of the mice in the donepezil positive drug group had an upward trend, but the difference was not statistically significant; the body weight of the mice in the low-dose cafoyl succinamide group was significantly increased ( p <0.01), approaching the level of the normal group. The above indicates that cafoyl succinamide has a certain improvement effect on the body weight loss of mice caused by modeling.

[0039] 2. Effect on the spontaneous alternation response rate of mice The results of the Y maze experiment to investigate the effect of cafoyl succinamide on the spontaneous alternation response rate of AD mice are shown in Figure 2 as follows.

[0040] The results showed that compared with the control group, the spontaneous alternation rate of the mice in the model group was significantly reduced ( p <0.01), indicating that the AD mouse model was successfully established, and D-galactose combined with AlCl3 could significantly damage the working memory ability of mice. Compared with the model group, the spontaneous alternation rates of the mice in the positive drug group and the cafoyl succinamide group were significantly increased ( p <0.001 or p <0.01). The above suggests that both the positive drug and cafoyl succinamide have obvious protective effects on the damage of the working memory ability of AD mice.

[0041] 3. Effect on the ability of mice to recognize new things The results of the effect of cafoyl succinamide on the ability of AD mice to recognize new things are shown in Figure 3 as follows.

[0042] The results showed that compared with the control group, the new object recognition index of the mice in the model group was significantly reduced ( p <0.05), indicating that the AD mouse model was successfully established, and D-galactose combined with AlCl3 could significantly damage the learning and memory ability of mice. Compared with the model group, the cognition of the mice in the positive drug group and the high-dose cafoyl succinamide group increased most significantly (p < 0.001 or p < 0.05). The above results indicate that both the positive drug and cafestolide have significant protective effects on the impairment of the ability to discriminate novel objects in AD mice.

[0043] 4. Effects on the escape latency and the number of platform crossings in mice The results of the Morris water maze experiment examining the effects of cafestolide on the escape latency and the number of platform crossings in AD mice are shown in Figure 4 the figure below.

[0044] The results showed that compared with the control group, the escape latency of the mice in the model group increased, and the number of platform crossings tended to decrease, but the difference was not statistically significant. Compared with the model group, the escape latency of the mice in the positive drug group decreased, and the number of platform crossings increased significantly ( p < 0.05), and the escape latency of the mice in the cafestolide group decreased, and the number of platform crossings increased significantly ( p < 0.01 or p < 0.05). The above results indicate that cafestolide can significantly improve the spatial memory ability of AD mice.

[0045] 5. Effects on the content of Aβ 40 and Aβ 42 in the mouse brain The results of the effects of cafestolide on the content of Aβ 40 and Aβ 42 in the brain tissue of AD mice are shown in Figure 5 the figure below.

[0046] The results showed that compared with the control group, the contents of Aβ 40 and Aβ 42 in the brain tissue of the mice in the model group increased significantly ( p < 0.001 or p < 0.01), suggesting that the pathogenesis of learning and memory impairment in AD mice induced by D-galactose combined with AlCl3 may be related to Aβ 40 and Aβ 42 . Compared with the model group, the contents of Aβ 40 and Aβ 42 in the brain tissue of the mice in the positive drug group and the cafestolide group decreased significantly ( p < 0.001 or p < 0.01). The results indicate that the protective effects of the positive drug and cafestolide on learning and memory impairment in AD mice may be related to their reduction of the contents of Aβ 40 and Aβ 42 in the brain tissue.

[0047] During the whole process of animal administration, no mice died, and there were no abnormalities in the appearance and behavior of the mice.

[0048] The various embodiments in this specification are described in a progressive manner. Each embodiment focuses on the differences from other embodiments. For the same or similar parts among the various embodiments, reference can be made to each other.

[0049] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but rather to the broadest scope consistent with the principles and novel features disclosed herein.

Claims

1. Use of cafoyl succinyl diamine in the preparation of medicaments for preventing and treating Alzheimer's disease, or in the preparation of health care products or functional foods for preventing and treating Alzheimer's disease.

2. The application according to claim 1, wherein Cafoyl succinyl diamine can significantly increase the spontaneous alternation response rate of Alzheimer's disease mice in the Y-maze experiment, that is, improve their working memory ability.

3. The application according to claim 1, characterized in that Cafoyl succinyl diamine can significantly increase the novel object recognition ability of Alzheimer's disease mice, that is, improve their learning ability.

4. The application according to claim 1, characterized in that, Cafoyl succinyl diamine can significantly reduce the escape latency of Alzheimer's disease mice in the Morris water maze experiment and increase the number of times of crossing the platform, that is, improve their spatial memory ability.

5. The application according to claim 1, wherein Caffeoyl succinamide can significantly reduce the levels of Aβ in the brain tissue of Alzheimer's disease mice 40 and Aβ 42 content.

6. A drug for preventing and treating Alzheimer's disease, characterized in that, It includes cafoyl succinyl diamine.

7. The drug according to claim 6, wherein, The dosage form is an orally administrable dosage form, external patch or injection dosage form permitted in pharmacy.

8. A health product for preventing and treating Alzheimer's disease, characterized in that, It includes cafoyl succinyl diamine.

9. A functional food for preventing and treating Alzheimer's disease, characterized in that, It includes cafoyl succinyl diamine.

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