Use of thienopyrimidinone derivatives for the preparation of medicaments for the treatment of alzheimer's disease
Thiophene pyrimidinone derivatives provide a highly effective treatment for Alzheimer's disease by inhibiting BChE activity, significantly improving cognitive deficits and overcoming the shortcomings of existing BChE inhibitors.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- LUDONG UNIVERSITY
- Filing Date
- 2026-03-13
- Publication Date
- 2026-06-26
AI Technical Summary
There is a lack of highly effective and safe butyrylcholinesterase (BChE) inhibitors for the treatment of Alzheimer's disease in the current technology, and existing drugs have not yet fully passed clinical validation.
The thienpyrimidinone derivative 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one was used as a BChE inhibitor to treat Alzheimer's disease by inhibiting BChE activity.
This thiophene pyrimidinone derivative exhibits highly effective inhibition of BChE, significantly improving cognitive deficits in AD models. The effect is dose-dependent and superior to existing drugs such as donepezil.
Smart Images
Figure CN121818655B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of thiophene pyrimidinone derivatives in the preparation of drugs for treating Alzheimer's disease. Background Technology
[0002] Alzheimer's disease (AD), a neurodegenerative disease, seriously threatens the health and quality of life of the elderly; however, highly effective treatments are still lacking. Butyrylcholinesterase (BChE) plays a crucial role in the pathological process of AD, and its unique distribution and functional characteristics make it an important target for AD treatment research.
[0003] BChE is a substrate-specific enzyme widely distributed in the central nervous system, found in areas closely related to learning and memory, such as the basal forebrain, hippocampus, and cortex. It is expressed in glial cells, neurons, and cerebral vascular endothelial cells, and also exhibits high activity in peripheral plasma, liver, and kidneys. In Alzheimer's disease (AD), BChE activity is significantly upregulated, driving disease progression through multiple mechanisms, including accelerating acetylcholine hydrolysis, promoting β-amyloid (Aβ) aggregation and fibrosis, and inducing neuroinflammation. Therefore, it has become an important target for AD treatment, and combined inhibition or targeted intervention strategies centered on BChE have shown good therapeutic potential. However, most related inhibitors are currently in the experimental stage, and their safety and efficacy have not been fully verified by clinical trials. Novel, highly effective, and safe BChE-related therapeutic drugs remain a direction that urgently needs to be explored in the field of AD treatment. Summary of the Invention
[0004] The purpose of this invention is to provide the application of thienylpyrimidinone derivatives in the preparation of drugs for treating Alzheimer's disease. 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one has a highly efficient inhibitory effect on BChE and can significantly improve cognitive function deficits in AD models, providing a novel and highly effective BChE-targeting candidate inhibitor for the treatment of AD.
[0005] To achieve the above objectives, this invention provides the use of a thiophene pyrimidinone derivative in the preparation of a drug for treating Alzheimer's disease. The thiophene pyrimidinone derivative is 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thiopheno[2,3-d]pyrimidin-4-one, with PubChem CID 17398588, and the structural formula is:
[0006] .
[0007] Preferably, Alzheimer's disease is a neurodegenerative disease caused by a deficiency in the cholinergic system.
[0008] Furthermore, cholinergic system dysfunction manifests as upregulation of butyrylcholinesterase activity, which compensatorily becomes the main cholinesterase in the brain.
[0009] Preferably, 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one treats Alzheimer's disease by inhibiting butyrylcholinesterase activity.
[0010] The present invention also provides a drug for treating Alzheimer's disease, comprising 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one and a drug carrier, wherein the drug carrier may be any known to those skilled in the art.
[0011] Therefore, the application of the above-mentioned thiophene pyrimidinone derivatives in the preparation of drugs for treating Alzheimer's disease has the following beneficial effects:
[0012] (1) In this invention, 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one has a highly efficient inhibitory effect on BChE, with a half-maximal inhibitory concentration (IC50) of 96.0 nM. Moreover, the inhibitory activity is significantly dose-dependent. It can directly block the hydrolytic effect of BChE on ACh, maintain the normal level of ACh in the synaptic cleft, and thus repair the cholinergic neurotransmission function. This provides a novel and highly efficient BChE-targeting candidate inhibitor for the treatment of AD.
[0013] (2) In this invention, 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one significantly improved the cognitive deficits in the AD model, and the effect showed a clear dose-dependent effect. It was superior to donepezil, a commonly used clinical drug, and provided a new potential intervention direction for solving the core symptoms of AD patients.
[0014] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description
[0015] Figure 1 The present invention describes the inhibitory effect of different concentrations of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one on BChE activity.
[0016] Figure 2This invention relates to the time and number of times mice in each group of the SAMP8 water maze experiment first crossed the platform position;
[0017] in Figure 2 In this context, 'a' represents the time when mice in each SAMP8 group first crossed the platform location. Figure 2 In the figure, b represents the number of times each group of mice in SAMP8 crossed the platform location. Detailed Implementation
[0018] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.
[0019] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.
[0020] In this invention, unless otherwise specified, the test materials and instruments are all conventional test materials in the field and can be purchased through commercial channels.
[0021] Example 1
[0022] The inhibitory effect of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one on BChE activity was determined by the Ellman colorimetric method.
[0023] The experiment was conducted in a 96-well microplate, with three replicates per well, and experimental, blank, and control groups were set up.
[0024] Experimental group: First, 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thiopheno[2,3-d]pyrimidin-4-one was prepared into a stock solution with dimethyl sulfoxide (DMSO), and then serially diluted with phosphate buffer (PBS) to obtain thiophenepyrimidinone derivative solutions with concentrations of 3 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1000 nM, 3000 nM, and 10000 nM. Add 30 μL of PBS buffer, 10 μL of thiophenepyrimidine ketone derivative solution, and 20 μL of BChE enzyme solution extracted from rat brain to a 96-well microplate sequentially (e.g., add 30 μL of PBS buffer, 10 μL of 3 nM thiophenepyrimidine ketone derivative solution, and 20 μL of BChE enzyme solution to the first well; add 30 μL of PBS buffer, 10 μL of 10 nM thiophenepyrimidine ketone derivative solution, and 20 μL of BChE enzyme solution to the second well; and so on). Pre-incubate at 37°C for 10 minutes to allow the thiophenepyrimidine ketone derivative to fully bind to the enzyme. Then add 60 μL of acetylthiocholine substrate to initiate the enzymatic reaction. After reacting for 15 minutes, add 80 μL of DTNB chromogenic reagent to terminate the reaction and develop color.
[0025] Blank group: Add 60 μL of PBS buffer to a 96-well microplate, followed by 60 μL of acetylthiocholine substrate. After 15 minutes, add 80 μL of DTNB chromogenic reagent and develop the color.
[0026] Control group: 40 μL of PBS buffer and 20 μL of BChE enzyme solution extracted from rat brain were added to a 96-well microplate in sequence. The plate was pre-incubated at 37°C for 10 minutes. Then, 60 μL of acetylthiocholine substrate was added to start the enzymatic reaction. After 15 minutes of reaction, 80 μL of DTNB chromogenic reagent was added to stop the reaction and develop the color.
[0027] After the color development stabilizes, the absorbance value A of each well is measured at a wavelength of 412 nm using a microplate reader, according to the formula:
[0028] Inhibition rate = [1-(A 实验 -A 空白 ) / (A 对照 -A 空白 )]×100%;
[0029] The inhibition rate of different concentrations of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thienro[2,3-d]pyrimidin-4-one on enzyme activity was calculated. A graph was plotted with the logarithm of the concentration (C) of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thienro[2,3-d]pyrimidin-4-one as the x-axis and the inhibition rate as the y-axis. The results are shown below. Figure 1 As shown, nonlinear regression analysis was performed using software such as GraphPad Prism to fit the dose-response curve, and then the concentration of the thiophene pyrimidine derivative (IC50 value) at an inhibition rate of 50% was calculated.
[0030] To ensure the reliability of the experimental results, the concentration of DMSO was controlled to not exceed 1% throughout the entire experimental process, and all experiments were independently repeated 3 times.
[0031] from Figure 1 As can be seen, with the increase of the concentration of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thieno[2,3-d]pyrimidin-4-one, the inhibition rate of BChE activity showed a significant upward trend, exhibiting a typical dose-dependent relationship. Using GraphPad Prism software, nonlinear regression analysis was performed on the data based on the Log (Inhibitor) vs. Response model. The IC50 of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thieno[2,3-d]pyrimidin-4-one against BChE was found to be 96.0 nM (95% confidence interval: 78.9–116.9 nM), and the coefficient of determination R of the fitted curve was [value missing]. 2 The value was 0.98 and the slope was 0.73, indicating a good fit of the data and that the results were statistically significant.
[0032] Example 2
[0033] The water maze test was used as a behavioral experiment to examine the effect of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one on improving cognitive impairment.
[0034] Laboratory animals: 9-month-old male SPF-grade rapidly aging mice (SAM mice), provided by the Laboratory Animal Facility of Ludong University.
[0035] Experimental grouping and administration regimen: SAMP8 mice were randomly divided into 4 groups of 10 mice each, including: SAMP8 control group; SAMP8-donepezil (Don) group (administered at a dose of 5 mg / kg, designated as SAMP8-Don group); SAMP8-2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one low-dose group (administered at a dose of 5 mg / kg, designated as SAMP8-thienpyrimidinone derivative low-dose group); and SAMP8-2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one high-dose group (administered at a dose of 15 mg / kg, designated as SAMP8-thienpyrimidinone derivative high-dose group). The drug was administered by gavage once daily for two consecutive months before a behavioral experiment was conducted.
[0036] All animals have free access to water and food. The environmental control standards are: room temperature 24±1℃, humidity 50%-60%, and 12 hours of light per day (8:00-20:00).
[0037] Experimental setup: Consists of a cylindrical metal pool (60cm high, 120cm in diameter), an automatic display and recording system, and a black platform (10cm in diameter). The pool walls are covered with black wallpaper. After filling with water, the water level is 1cm above the platform, and the water temperature is maintained at 23±1℃. The experiment was conducted in a dimly lit, soundproof room. Fixed shape markers were affixed to the surrounding walls to ensure that mice could not locate the platform using sight, hearing, or smell, thus testing spatial memory ability. The pool was divided into four quadrants (I-IV), with the platform fixed at the center of quadrant II.
[0038] Experimental Procedure: For the first 6 days, a navigational training experiment was conducted: each mouse received two platform-finding training sessions daily, entering the water headfirst from two semi-random locations (e.g., the midpoint between quadrants I and II, or the midpoint between quadrants II and III). The time to find the platform (latency period) was recorded, and the average of the two readings was taken as the result for that day. If the platform was not found within 90 seconds, the latency period was recorded as 90 seconds. After finding the platform, the mouse was allowed to remain on it for 30 seconds to reinforce memory. After the navigational training, an exploration experiment was conducted: the platform was removed, and the mice swam freely for 90 seconds. The time to first cross the platform location and the number of times the mice crossed the platform were recorded.
[0039] Data Analysis: Experimental results are expressed as mean ± SEM. Statistical analysis was performed using SPSS 17.0 software, and graphs were plotted using GraphPad Prism 5 software. One-way ANOVA was used for the water maze exploration experiment, and post-hoc NK tests were used for comparisons between groups. p <0.05 indicates a statistically significant difference.
[0040] The results of the water maze exploration experiment are as follows Figure 2 As shown, from Figure 2 The results show that the time to first cross the platform in the SAMP8 control group was 65.9±7.5 seconds, and the number of crossings was 1.1±0.3; the time and number of crossings in the SAMP8-Don group were 43.2±3.6 seconds and 2.7±0.3, respectively; the time and number of crossings in the low-dose SAMP8-thiaphenidone derivative group were 35.6±4.3 seconds and 3.4±0.4, respectively; and the time and number of crossings in the high-dose SAMP8-thiaphenidone derivative group were 26.4±5.8 seconds and 4.8±0.7, respectively. Compared with the SAMP8 control group, the time to first cross the platform was significantly shorter in the SAMP8-Don group. p <0.01, the time was significantly shortened in the low-dose group of SAMP8-thiophenepyrimidine derivative ( p <0.01, the time was significantly shortened in the high-dose group of SAMP8-thiophenepyrimidine derivative (SAMP8-thiophenepyrimidine derivative). p <0.001); Number of platform crossings: The number of platform crossings in the SAMP8-Don group increased significantly ( p <0.05); the number of platform crossings was significantly increased in the low-dose group of SAMP8-thiophenepyrimidine derivative ( p <0.01); the high-dose group of SAMP8-thiophenepyrimidine derivatives showed a highly significant increase in the number of platform crossings ( p <0.001).
[0041] The results of the water maze test showed that both low and high doses of 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one significantly improved the learning and memory abilities of SAMP8 mice in a dose-dependent manner. The number of platform crossings and the latency to the first platform crossing were better in the low-dose group and the high-dose group of SAMP8-thienpyrimidinone derivative than in the SAMP8-Don group.
[0042] Therefore, this invention utilizes the above-mentioned thiophene pyrimidinone derivative in the preparation of drugs for treating Alzheimer's disease. 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thiopheno[2,3-d]pyrimidin-4-one has a highly efficient inhibitory effect on BChE and can significantly improve cognitive function deficits in AD models, providing a novel and highly effective BChE-targeting candidate inhibitor for the treatment of AD.
[0043] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.
Claims
1. The application of thiophene pyrimidinone derivatives in the preparation of drugs for treating Alzheimer's disease, characterized in that: The thienylpyrimidinone derivative is 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thienro[2,3-d]pyrimidin-4-one, with the following structural formula: 。 2. The use of the thiophene pyrimidinone derivative according to claim 1 in the preparation of drugs for treating Alzheimer's disease, characterized in that: Alzheimer's disease is a neurodegenerative disease caused by a deficiency in the cholinergic system.
3. The use of the thiophene pyrimidinone derivative according to claim 2 in the preparation of drugs for treating Alzheimer's disease, characterized in that: Cholinergic system dysfunction manifests as upregulation of butyrylcholinesterase activity, which compensatorily becomes the main cholinesterase in the brain.
4. The use of the thiophene pyrimidinone derivative according to claim 1 in the preparation of drugs for treating Alzheimer's disease, characterized in that: 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thien-2-yl)thieno[2,3-d]pyrimidin-4-one treats Alzheimer's disease by inhibiting butyrylcholinesterase activity.
5. A drug for treating Alzheimer's disease, characterized in that: Includes 2-[(4-methylpiperidin-1-yl)methyl]-3-(tetrahydrofuran-2-ylmethyl)-5-(thiophen-2-yl)thiopheno[2,3-d]pyrimidin-4-one and drug carriers.