Application of N1, N10-dicaffeoyl spermidine in preparation of medicine for preventing and treating Alzheimer disease
By using N1,N10-dicafenimide as the main ingredient, the existing AD therapeutic drug targets are single, significant side effects and high cost are solved, and the memory capacity of AD mice is significantly improved and the inflammatory factors and Aβ40/42 levels are achieved.
Patent Information
- Application Number
- CN202510558375.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-29
- Publication Date
- 2025-06-20
AI Technical Summary
The existing Alzheimer's disease (AD) therapeutic drugs have problems such as single targets, inability to reverse the pathological process, significant side effects and high costs. It is urgent to develop new drugs that have both multi-target regulation, neuroprotection and better safety.
N1,N10-dicafenimide was used as the main active ingredient, and it was found that it significantly improved the effect of AD through previous animal experiments, and it was developed as a drug for preventing and treating AD.
N1,N10-dicafenimide significantly improves the working memory, spatial memory and learning ability of AD mice, reduces brain tissue inflammatory factors and Aβ40/42 levels, proving that it has the effect of preventing and treating AD.
Smart Images
Figure CN120168447A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedical technologies, and more specifically relates to the use of N1,N10-dicaffeoyl spermidine in the preparation of drugs for preventing and treating Alzheimer's disease. Background Art
[0002] Alzheimer's Disease (AD) is the most common neurodegenerative disease globally, accounting for approximately 60%-70% of dementia cases in the elderly. Statistics from the World Health Organization show that the number of AD patients globally has exceeded 55 million and is expected to increase to 139 million by 2050. Its incidence rate increases exponentially with the aggravation of population aging. AD not only leads to the progressive loss of cognitive function and the degradation of living ability in patients, but also imposes a heavy burden on the social medical system. The global medical costs related to AD exceeded $1.5 trillion in 2023. The pathological mechanism of AD is complex, and its core features include β-amyloid (Aβ) deposition, hyperphosphorylation of Tau protein, cholinergic system defects, and neuroinflammation and oxidative stress, etc.
[0003] Currently, the clinical treatment drugs for AD mainly include acetylcholinesterase (AChE) inhibitors, such as donepezil and galantamine, which improve cognition in the short term by inhibiting AChE to increase the synaptic ACh concentration, but cannot delay the disease progression, and are accompanied by gastrointestinal side effects and decreasing efficacy. NMDA receptor antagonists, such as memantine, regulate glutamate excitotoxicity, but the single-drug effect is limited and often needs to be used in combination with AChE inhibitors. Aβ-targeted monoclonal antibodies: such as Lecanemab, can remove Aβ plaques in the brain, but there are risks such as cerebral edema and microbleeding, and the efficacy for mid- and late-stage patients is not significant. Tau protein inhibitors: such as antisense oligonucleotides, are still in the clinical trial stage, and their long-term safety remains to be verified. Existing drugs generally have problems such as single targets, inability to reverse the pathological process, significant side effects, and high costs, and there is an urgent need to develop new drugs with multi-target regulation, neuroprotection, and better safety.
[0004] Natural products have become an important direction for AD drug research and development due to their advantages such as multi-target synergistic potential, blood-brain barrier penetrability, and long-term application safety. At the same time, the natural product library is rich in phenolic acids, alkaloids, and terpene compounds, and their unique structures can provide templates for designing novel AChE / Aβ dual-target inhibitors.
[0005] The compound N1,N10-dicaffeoyl of the present invention is an active compound isolated from traditional Chinese medicines such as wolfberry. Currently, there are few reports on the biological activities of N1,N10-dicaffeoyl spermidine, and there is no relevant research on the use of N1,N10-dicaffeoyl spermidine for preventing and treating Alzheimer's disease. Summary of the Invention
[0006] In view of this, the present invention provides the use of N1,N10-dicaffeoylspermidine in the preparation of drugs for preventing and treating Alzheimer's disease. Through preliminary animal experiments, the present invention found that N1,N10-dicaffeoylspermidine has a significant effect on improving Alzheimer's disease. Further, developing it into a drug that can be used to prevent and treat Alzheimer's disease will have great clinical application value.
[0007] N1,N10-dicaffeoylspermidine, with the molecular formula C 25 H 31 N3O6, a molecular weight of 469.22, and the full English name of (E)-3-(3,4-dihydroxyphenyl)-N-(3-((4-((E)-3-(3,4-dihydroxyphenyl)acrylamido)butyl)amino)propyl)acrylamide, abbreviated as N1,N3-bis(caffeoyl)spermidine or N1,N3-di-caffeoylspermidine. Its chemical structural formula is shown in Formula (1): Formula (1) To achieve the above object, the present invention adopts the following technical solutions: The use of N1,N10-dicaffeoylspermidine in the preparation of drugs for preventing and treating Alzheimer's disease (AD).
[0008] Further, N1,N10-dicaffeoylspermidine can significantly increase the spontaneous alternation response rate of AD mice in the Y-maze experiment, that is, improve their working memory ability.
[0009] Further, N1,N10-dicaffeoylspermidine can significantly increase the novel object recognition ability of AD mice, that is, improve their learning ability.
[0010] Further, N1,N10-dicaffeoylspermidine can significantly reduce the escape latency of AD mice in the Morris water maze experiment and increase the number of times of crossing the platform, that is, improve their spatial memory ability.
[0011] Further, N1,N10-dicaffeoylspermidine can significantly reduce the contents of inflammatory factors TNF-α, IL-1β and IL-6 in AD mice.
[0012] Further, N1,N10-dicaffeoylspermidine can significantly reduce Aβ 40 and Aβ 42 contents in the brain tissue of AD mice.
[0013] A drug for preventing and treating Alzheimer's disease, characterized by comprising N1,N10-dicaffeoyl spermidine.
[0014] Furthermore, the dosage form is an orally administrable dosage form, an external patch or an injection dosage form permitted in pharmacy.
[0015] Through the above technical solutions, it can be seen that compared with the prior art, the beneficial effects achieved by the present invention are as follows: within the dose range of 10-25 mg / kg administered to experimental animals, N1,N10-dicaffeoyl spermidine can significantly improve the working memory, spatial memory and learning ability of D-galactose combined with aluminum trichloride-induced AD mice, and significantly reduce the contents of inflammatory factors TNF-α, IL-1β and IL-6 in the brain tissue of AD mice. At the same time, it significantly reduces Aβ 40 and Aβ 42 levels. The above confirms that N1,N10-dicaffeoyl spermidine 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
[0016] 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, other drawings can be obtained according to the provided drawings without creative efforts.
[0017] Figure 1 Effect of N1,N10-dicaffeoyl spermidine on the body weight of AD mice (C is the normal group, M is the model group, P is the positive drug donepezil group, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the normal group, ## p <0.01; compared with the model group, ** p <0.01, *** p <0.001) Figure 2 Effect of N1,N10-dicaffeoyl spermidine 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, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the normal group, ## p <0.01; compared with the model group, ** p <0.01, *** p <0.001) Figure 3 To investigate the effect of N1,N10-dicaffeoyl spermidine on the ability of AD mice to discriminate novel objects in the novel object recognition experiment (C is the normal group, M is the model group, P is the positive drug donepezil group, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the normal group, # p <0.05; compared with the model group, ** p <0.01, *** p <0.001) Figure 4 To investigate the effect of N1,N10-dicaffeoyl spermidine 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, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the model group, * p <0.05, ** p <0.01) Figure 5 To investigate the content of inflammatory factors TNF-α, IL-1β, and IL-6 in the brain tissue of AD mice by N1,N10-dicaffeoyl spermidine (C is the normal group, M is the model group, P is the positive drug donepezil group, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the normal group, ### p <0.001; compared with the model group, * p <0.05, ** p <0.01, *** p <0.001) Figure 6 To investigate the effect of N1,N10-dicaffeoyl spermidine 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, NNDCP-H is the high-dose group of N1,N10-dicaffeoyl spermidine, NNDCP-L is the low-dose group of N1,N10-dicaffeoyl spermidine; compared with the normal group, ## p <0.01, ### p <0.001; compared with the model group, ** p <0.01, *** p <0.001) Detailed implementation manners
[0018] 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 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.
[0019] In the following examples, N1,N10-dicaffeoyl spermidine used is the compound shown in the above formula (1), and can be obtained through commercial purchase or experimental self-preparation.
[0020] The medicaments required for the present invention are conventional experimental medicaments and 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
[0021] A drug for preventing and treating Alzheimer's disease, with an oral dosage form, including N1,N10-dicaffeoyl spermidine, wherein the single application dose of N1,N10-dicaffeoyl spermidine is 10 mg / kg. Example 2
[0022] A drug for preventing and treating Alzheimer's disease, with an oral dosage form, including N1,N10-dicaffeoyl spermidine, wherein the single application dose of N1,N10-dicaffeoyl spermidine is 25 mg / kg. Example 3
[0023] A drug for preventing and treating Alzheimer's disease, with an injection dosage form, including N1,N10-dicaffeoyl spermidine, wherein the single application dose of N1,N10-dicaffeoyl spermidine is 10 mg / kg. Example 4
[0024] A drug for preventing and treating Alzheimer's disease, with an external patch dosage form, including N1,N10-dicaffeoyl spermidine, wherein the single application dose of N1,N10-dicaffeoyl spermidine is 25 mg / kg.
[0025] The following animal experiments further illustrate the effects of the above Examples 1 to 4: I. Animal experiment design An Alzheimer's disease (AD) mouse model was induced by D-galactose combined with aluminum trichloride (AlCl3). After intragastric administration of N1,N10-dicaffeoyl spermidine, through mouse body weight, Y-maze experiment, novel object recognition experiment, Morris water maze experiment, Aβ 40 and Aβ 42Evaluate the preventive and therapeutic effects of N1,N10-dicaffeoyl spermidine on Alzheimer's disease.
[0026] II. Experimental procedures 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). They were housed in an SPF-grade barrier environment with a 12-hour light and 12-hour dark cycle and had free access to food and water.
[0027] 2. Experimental methods An Alzheimer's disease (AD) mouse model was induced by D-galactose combined with aluminum chloride (AlCl3). After 3 days of adaptive feeding, 48 mice were randomly divided into 5 groups according to their body weight: normal group (saline), model group (saline), donepezil positive drug group (5 mg / kg), N1,N10-dicaffeoyl spermidine low-dose group (NNDCP-L, 10 mg / kg / d), and N1,N10-dicaffeoyl spermidine high-dose group (NNDCP-H, 25 mg / kg / d), with 10 mice in each group. Donepezil and N1,N10-dicaffeoyl spermidine were dissolved in saline and administered by gavage at a dose of 10 ml / kg. Mice were subcutaneously injected with D-galactose (200 mg / kg) at a fixed time point every day, and simultaneously gavaged with 150 mg / kg AlCl3 to induce the AD model (the success of the model was judged by behavioral experiments 5 weeks after mouse modeling). Mice in the normal group were subcutaneously injected and gavaged with an equal volume of saline; gavage administration was performed 4 hours later. The above modeling and gavage were carried out once a day for 10 consecutive weeks of modeling and drug administration.
[0028] 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 N1,N10-dicaffeoyl spermidine on the working memory ability of AD mice, and the spontaneous alternation response rate of each mouse was calculated for evaluation.
[0029] 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 N1,N10-dicaffeoyl spermidine on the learning and memory ability of AD mice, and the preference index of each mouse was calculated for evaluation.
[0030] 5. Morris water maze experiment After 10 weeks of drug administration to each group of mice, the Morris water maze experiment (continuously measured for 5 days) was used to investigate the effect of N1,N10-dicaffeoyl spermidine 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.
[0031] 6, TNF-α, 1L-1β, 1L-6, Aβ 40 and Aβ 42 Content determination After 10 weeks of drug administration and completion of behavioral tests in each group of mice, after taking serum, the mice were decapitated and sacrificed, and the whole brain was taken by decapitation on ice, immediately frozen in liquid nitrogen, and 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 TNF-α, 1L-1β, 1L-6, Aβ 40 and Aβ 42 Content.
[0032] 7. Data processing The data processing of this invention uses ImageJ and Graphpad Prism 9.5 for data analysis and statistics. The obtained results are all expressed as mean ± standard deviation (X±S). One-way ANOVA is used for data processing of the inter-group difference comparison. The difference is considered statistically significant with p<0.05 as the standard.
[0033] III. Experimental results 1. Effect on mouse body weight The results of the effect of N1,N10-dicaffeoyl spermidine on mouse body weight are shown in Figure 1 the figure.
[0034] After 10 weeks of drug administration in 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 group of N1,N10-dicaffeoyl spermidine was significantly increased ( p <0.001 or p <0.01), approaching the normal group level. The above indicates that N1,N10-dicaffeoyl spermidine has a certain improvement effect on the weight loss of mice caused by modeling.
[0035] 2. Effect on the spontaneous alternation response rate of mice The results of the Y maze experiment examining the effect of N1,N10-dicaffeoyl spermidine on the spontaneous alternation response rate of AD mice are shown in Figure 2 the figure.
[0036] 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 mouse model of Alzheimer's disease 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 rate of mice in the positive drug group and the N1,N10-dicaffeoyl spermidine group increased significantly ( p <0.001 or p <0.01). The above results suggest that both the positive drug and N1,N10-dicaffeoyl spermidine have obvious protective effects on the damage of working memory ability in AD mice.
[0037] 3. Effects on the ability of mice to recognize new things The results of the effects of N1,N10-dicaffeoyl spermidine on the ability of AD mice to recognize new things are shown in Figure 3 the figure.
[0038] The results showed that compared with the control group, the cognitive index of new objects in the model group of mice was significantly decreased ( p <0.05), indicating that the mouse model of Alzheimer's disease 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 cognitive ability of mice in the positive drug group and the N1,N10-dicaffeoyl spermidine group increased most significantly ( p <0.001 or p <0.01). The above results suggest that both the positive drug and N1,N10-dicaffeoyl spermidine have obvious protective effects on the damage of the ability of AD mice to distinguish new things.
[0039] 4. Effects on the escape latency and the number of platform crossings of mice The results of the Morris water maze experiment examining the effects of N1,N10-dicaffeoyl spermidine on the escape latency and the number of platform crossings of AD mice are shown in Figure 4 the figure.
[0040] The results showed that compared with the control group, the escape latency of 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 mice in the positive drug group decreased, and the number of platform crossings increased significantly ( p <0.05), and the escape latency of mice in the N1,N10-dicaffeoyl spermidine group decreased, and the number of platform crossings increased significantly ( p <0.01). The above results indicate that N1,N10-dicaffeoyl spermidine can significantly improve the spatial memory ability of AD mice.
[0041] 5. Effects on the contents of inflammatory factors TNF-α, IL-1β, and IL-6 in the brain tissue of mice The results of the effects of N1,N10-dicaffeoyl spermidine on the contents of inflammatory factors TNF-α, IL-1β, and IL-6 in the brain tissue of AD mice are shown inFigure 5 as shown
[0042] The results showed that, compared with the control group, the contents of TNF-α, IL-1β, and IL-6 in the brain tissues of mice in the model group were significantly increased ( p <0.001), suggesting that D-galactose combined with AlCl3 induced an increase in inflammatory factors in AD mice. Compared with the model group, the contents of TNF-α, IL-1β, and IL-6 in the brain tissues of mice in the positive drug group and the N1,N10-dicaffeoyl spermidine group were significantly decreased ( p <0.001, p <0.01 or p <0.05). The above suggests that the positive drug and N1,N10-dicaffeoyl spermidine can significantly reduce the levels of inflammatory factors in the brain tissues of AD mice.
[0043] 6. Effects on the contents of Aβ 40 and Aβ 42 in the brain tissues of mice The effects of N1,N10-dicaffeoyl spermidine on the contents of Aβ 40 and Aβ 42 in the brain tissues of AD mice are shown in Figure 6 as shown
[0044] The results showed that, compared with the control group, the contents of Aβ 40 and Aβ 42 in the brain tissues of mice in the model group were significantly increased ( p <0.001 or p <0.01), suggesting that the pathogenesis of learning and memory impairment induced by D-galactose combined with AlCl3 in AD mice 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 tissues of mice in the positive drug group and the N1,N10-dicaffeoyl spermidine group were significantly decreased ( p <0.001 or p <0.01). The results showed that the protective effects of the positive drug and N1,N10-dicaffeoyl spermidine 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 tissues.
[0045] During the whole process of drug administration to animals, no mice died, and there were no abnormal conditions in the appearance and behavior of the mice.
[0046] 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.
[0047] The foregoing 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 readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. Application of N1,N10-dicaffeoylspermidine in the preparation of drugs for preventing and treating Alzheimer's disease.
2. The use according to claim 1, characterized in that: N1,N10-dicaffeoylspermidine can significantly increase the spontaneous alternation response rate of Alzheimer's disease mice in the Y-maze test, that is, improve their working memory ability.
3. The use according to claim 1, characterized in that: N1,N10-dicaffeoylspermidine can significantly increase the ability of Alzheimer's disease mice to recognize new objects, that is, improve their learning ability.
4. The use according to claim 1, characterized in that: N1,N10-dicaffeoylspermidine can significantly reduce the escape latency of Alzheimer's disease mice in the Morris water maze test and increase the number of platform crossings, that is, improve their spatial memory ability.
5. The use according to claim 1, characterized in that: N1,N10-dicaffeoylspermidine significantly reduces Aβ in brain tissue of Alzheimer's disease mice 40 and Aβ 42 content.
6. The use according to claim 1, characterized in that: N1,N10-dicaffeoylspermidine can significantly reduce the levels of inflammatory factors TNF-α, 1L-1β and 1L-6 in the brain tissue of Alzheimer's disease mice.
7. A drug for preventing and treating Alzheimer's disease, characterized in that: Including N1,N10-dicaffeoylspermidine.
8. The drug according to claim 7, characterized in that The dosage form is a pharmaceutically acceptable oral dosage form, external patch or injection dosage form.