CN115813917A - A compound capable of rapidly improving cognitive function and its application - Google Patents

Abstract

The invention belongs to the field of medicines, and discloses a compound for improving cognitive impairment, which can rapidly increase the activity of neurons and enhance the plasticity of nerve synapses, thereby rapidly improving cognitive functions.

Description

A compound capable of rapidly improving cognitive function and its application

technical field

The invention relates to the technical field of medicine preparation, and more specifically relates to a compound capable of rapidly improving cognitive function and its application.

Background technique

Alzheimer's disease (AD) causes memory and other cognitive decline and is responsible for more than 60% of dementia cases. Most of the existing marketed drugs are aimed at a single target as brain metabolism improving agents or neuroprotective agents, but their effects are minimal. In recent years, with the gradual deepening of pharmacological activity research, the use of hallucinogens to improve the cognitive function of AD patients has gradually increased, and it shows a considerable development trend in alleviating AD symptoms and cognitive impairment. Since Alzheimer's disease is currently incurable, improving its main symptom, cognitive impairment, has become the main treatment option, and related drugs have also become a rigid demand.

Therefore, how to provide a compound that improves cognitive function is an urgent problem to be solved by those skilled in the art.

Contents of the invention

In view of this, the present invention provides a compound capable of rapidly improving cognitive function, which can rapidly improve spatial memory ability and cognitive function.

In order to achieve the above object, the present invention adopts the following technical solutions:

The chemical formula of the compound with rapid improvement of cognitive function provided by the present invention is:

The medicine dimethyltryptamine in the present invention is the first class of psychotropic drugs and belongs to tryptamine hallucinogens. Structurally similar to neurotransmitters, serotonin and other hallucinogens psilocybin. Dimethyltryptamine was originally found mainly in plants and in trace amounts in mammalian brains. The drug can then also be synthesized in the laboratory.

The compounds of the present invention that can rapidly improve cognitive function and improve Alzheimer's disease can be prepared into preparations such as injections and sprays.

The preparation for rapidly improving memory and cognitive ability and improving senile dementia described in the present invention can also add pharmaceutically acceptable conventional auxiliary materials.

Indications for the above compounds: memory loss; impairment of cognitive function; Alzheimer's disease.

The pharmacological properties of the present invention are as follows: the activity of neurons can be rapidly increased, the plasticity of nerve synapses can be enhanced, and cognitive functions can be rapidly improved.

The compound described in the present invention can be prepared into a pharmaceutically acceptable salt, and the compound is used in the treatment of neurons damaged by neurotoxin and has obvious repairing effect.

In the present invention, the raw material medicine capable of rapidly improving cognitive function and neuroprotection is dissolved in physiological saline to 0.25-1 mg/ml.

It can be seen through the above-mentioned technical solution that, compared with the prior art, the beneficial effects of the present invention are as follows:

The invention discloses a new application of a dimethyltryptamine compound in the preparation of neuroprotective drugs, which can rapidly improve cognitive impairment and is used for preparing neuroprotective drugs.

Description of drawings

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 that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only It is an embodiment of the present invention, and those skilled in the art can also obtain other drawings according to the provided drawings without creative work.

Figure 1 shows the effects of drugs that rapidly improve cognitive function on LTP in the mouse hippocampus.

Figure 2 shows the effects of drugs that rapidly improve cognitive function on calcium activity and neuronal ATP in the hippocampus of mice.

Detailed ways

The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

The drugs used in the examples were approved by the Department of Health of the Hong Kong Special Administrative Region and purchased from Sigma Corporation of the United States. The drugs used in the comparative examples were purchased from Sigma Corporation of the United States.

In the embodiment, the raw material medicine that can improve cognitive function and neuroprotection is dissolved in physiological saline to 1 mg/ml.

This comparative example is donepezil, and the preparation method of this comparative example is: dissolving the raw material of donepezil in medicinal saline to 0.5 mg/ml.

The following tests show that the compound of the chemical formula of the present invention has rapid improvement of cognitive function and neuroprotective effect, and can be used in medicines for treating or preventing senile dementia.

Example 1

(1) Test material

The experimental drug is the compound with rapid improvement of cognitive function and neuroprotective effect in the embodiment and donepezil as the comparative example.

The experimental animals were Alzheimer's disease transgenic mice (3TG-AD; 5×FAD), purchased from Jackson Laboratory in the United States; SPF grade C57BL/6 mice were purchased from the Comparative Medicine Research Center of the University of Hong Kong, license number: 5345- 20. All mice were raised in an environment with a temperature of 23±1°C and a humidity of about 55%. Keep the lighting time for 12 hours, irradiate the natural light evenly, 6/cage, the mice can take food and water freely.

(2) Test method

Grouping and administration of mice: mice were divided into control group (C57BL/6), model group (3TG-AD or 5×FAD mice), and treatment group (compound of the present invention). The drugs in the treatment group were dissolved in physiological saline to an appropriate concentration, and a single dose was administered intraperitoneally to the mice. The dose for the example group was 12 mg/kg, and the dose for the control group was 5 mg/kg.

Animal behavior test: mice were injected intraperitoneally with the drug of the present invention for about 30 minutes to perform a Y maze test to detect the effect of the drug of the present invention on rapidly improving the spatial memory and cognitive ability of mice.

Y maze: Y maze consists of three identical arms, each arm has an angle of 120 degrees, each arm measures 30cm*8cm*15cm (length*width*height), and there is a movable partition in the center . The three arms of each Y-maze were randomly assigned: the new arm, the starting arm, and the other arm. Novel arm: blocked in the first phase of the experiment, that is, the training phase, and opened in the second phase of the test phase; starting arm: the arm where the mice entered the maze. The start arm and other arms were kept open throughout the experiment, allowing animals free access. After each training or testing session, remove animal excrement with alcohol to prevent residual animal odor disturbance.

The Y maze test of animal spatial recognition and memory ability includes two stages, with an interval of 1 hour. The first stage is the acquisition stage. The novel arm is closed, and the animal is allowed to explore freely in the other two arms for 5 minutes. A second experiment (recall phase) was performed 1 hour later, with all arms opened and the animals moving freely in three arms for 5 min. The time and distance the animals explored in each arm were recorded using the SMART video tracking system.

Mouse hippocampal long-term potentiation (LTP) test: the mouse to be tested was quickly decapitated, and the scalp was cut to remove the skull and dura mater. The whole brain was quickly removed and placed in artificial cerebrospinal fluid (ACSF) saturated with 95% O ₂ and 5% CO ₂ at 0-4° C. for slight cooling. Cut off the cerebellum and 1/3 of the forebrain, divide the brain into two along the midline, and fix the brain tissue containing the hippocampus on the loading bath dish with glue. Brain slices with a thickness of 350 μm were sectioned coronally with a vibrating microtome. The brain slices were placed in an incubation tank immersed in a nylon mesh under the liquid surface, filled with mixed gas continuously, and incubated in a constant temperature water bath at 34°C for 0.5h, and then incubated at room temperature for use. In the laboratory, the recording point was placed on the vertebral cell layer in the CA1 area under the operating microscope, and the recorded signal can be displayed on the memory oscilloscope and the computer display screen at the same time. The changes of excitatory postsynaptic potential (fEPSP) and its slope were monitored every 20s for 60min.

Calcium activity in mouse hippocampal cells and ATP level in hippocampal neurons: Optical fiber photometry was used to detect calcium signals in the mouse hippocampus and ATP levels in neurons. Anesthetized mice were fixed on a stereotaxic apparatus. Cut the mouse scalp along the midline, clean the soft tissue on the surface of the mouse, and position the skull surface: refer to the mouse atlas bregma: AP, -1.85mm; ML, ±1.10mm; DV, 2.04mm. A certain amount of virus is sucked up by a micro-syringe pump. The mice were craniotomized, and after 5 minutes of rest, the virus (AAV9-syn-RCaMP; AAV9-syn-ATP1.0) was injected with a microinjection pump at a rate of 50nL/min, and the injection volume was 200nL. After the injection is completed, withdraw slowly after stopping for 10 minutes. Clean up the injection site and surrounding debris and suture the scalp. After the mice woke up, they were put back into the cage. After the virus injection, continue to feed for 2 weeks, implant the optical fiber in the hippocampus of the mouse, and continue to feed for 1 week. After the virus infects neurons and fully expresses Ca ²⁺ fluorescent indicator protein (RCaMP) and ATP fluorescent protein (ATP1.0), Record the Ca ²⁺ level in the hippocampus and the ATP level in the hippocampal neurons after the injection of the drug in the awake and free state of the mice. At the same time, the camera is used to collect the behavioral data synchronously, and the recording lasts for about 30 minutes.

Data processing method: The experimental data are expressed as mean ± standard deviation (mean ± S.E.). Statistically significant differences According to the one-way ANOVA analysis of Graphpadprism 9.0 software, the differences between groups were tested (Tukey's test was performed afterwards), and P<0.05 was considered to be significant and statistically significant.

(3) Experimental results:

Y maze test results: the experimental results of each group of mice in the new arm are shown in Table 1, the treatment group is the drug treatment group of the present invention, and the test is carried out 30 minutes after the single dose injection administration, the time of the mice in the new arm significantly increased (P<0.05).

Table 1 Effects of fast-improving cognitive function drugs on mouse Y maze experiment

group number of animals New different arm time (%) P value Blank control group 10 24.85±2.781 model group 10 5.48±2.98 P<0.0001 Example group 10 17.86±2.47 P<0.01 Comparative group 10 2.54±2.00 P＞0.05

Note: Data are expressed as mean ± standard deviation (n = 10/group). P<0.01, P<0.0001 means extremely significant.

Mouse hippocampal long-term potentiation (LTP) test: the treatment group (the drug of the present invention) took brain slices to test the mouse hippocampal LTP within 1 hour after the mice were injected with a single dose, and the results showed that the treatment group significantly enhanced the excitement of the AD mice Sexual postsynaptic potential (fEPSP) (P<0.0001) (Figure 1), the average fEPSP recorded 50-60 minutes after theta pulse stimulation (TBS), treatment drugs also significantly increased fEPSP in AD mice, and the treatment group significantly higher than that of the blank control group, indicating that the drug of the present invention can rapidly change LTP and improve the synaptic function of the mouse hippocampus.

As shown in Figure 1, Figure 1 shows the effect of drugs that rapidly improve cognitive function on LTP in the hippocampus of mice. The figure shows the excitatory postsynaptic potential (fEPSP) of the hippocampus of mice in each group 15 minutes before theta pulse stimulation and 60 minutes after the stimulation.

Table 2 Effects of rapidly improving cognitive drugs on mouse hippocampal LTP

Note: Data are mean fEPSP recorded 50-60 minutes after theta pulse stimulation (TBS), expressed as mean ± standard deviation (n = 10/group). P<0.01, P<0.0001 means extremely significant.

Calcium ions in mouse hippocampal cells and ATP levels in hippocampal neurons: The optical fiber system was used to record the changes of calcium activity and ATP in the hippocampus of mice in each group in the state of waking and free activity. The results showed that after a single injection of the drug of the present invention in mice, the treatment group obviously induced the change of Ca ²⁺ (Fig. 2), and the calcium signal increased rapidly from 5 minutes after the injection of the drug of the present invention, and the Ca in the hippocampus of the mice of the treatment group ²⁺ signal increased significantly (P<0.05). It shows that the medicine of the present invention can significantly increase the activity of mouse hippocampal neurons.

The optical fiber system was used to record the ATP level of mouse hippocampal neurons after a single injection of the drug of the present invention, and the results showed that compared with the control group (injection of normal saline), the drug of the present invention can significantly increase the release of ATP in the mouse hippocampus (P<0.05).

Figure 2 shows the effects of drugs that rapidly improve cognitive function on calcium activity and neuronal ATP in the hippocampus of mice. AD mice were injected with physiological saline and the drug of the present invention, respectively, and the calcium activity and neuronal ATP signal in the hippocampus of the mice were recorded and compared (n=4-5). P<0.05 represents a significant difference.

Example 2

(1) Test material

The experimental drug is the compound with rapid improvement of cognitive function and neuroprotective effect in the examples. The primary hippocampal neurons of mice were used in this experiment, and the mice were provided by the Center for Comparative Medicine, University of Hong Kong, license number: 5369-20.

(2) Test method

The protective effects of the drugs in the above examples on Aβ _25-35- induced damage to primary hippocampal neurons of mice were investigated. Cells were seeded in a 96-well plate at a density of 8×10 ³ cells/well, cultured in a 5% CO ₂ , 37°C constant temperature incubator for 7 days, and the cell state was observed _. Modeling treatment was performed on the cells at _-35 , and the blank group was not treated, and they were co-incubated for 24 hours. Add CCK-8 solution to the 96-well plate in the dark, and incubate for 2 hours in the dark, then detect its absorbance at 450 nm, and calculate the change of cell viability.

The survival rate of each group was calculated with the survival rate of the blank group as 100%. Experimental data are expressed as mean ± standard error (mean ± S.E.). Statistically significant differences According to the one-way ANOVA analysis of GraphPadprism 9.0 software, the differences between groups were tested (Tukey's test was performed afterwards), and P<0.05 was considered to be significant and statistically significant.

(3) Test results

Table 3 shows the effects of the drugs with rapid improvement of cognitive function and neuroprotective effect on the viability of mouse primary hippocampal neurons in the examples. Compared with the blank group, Aβ _25-35 (final concentration 10 μM) decreased the cell viability to 70.53% (P<0.01), and when the administration concentration was 2 μM, the cell viability in the example group increased significantly (P<0.001). Drug preconditioning that rapidly improves cognitive function has a significant neuroprotective effect on Aβ _25-35- induced cell damage.

Table 3 Neuroprotective effects of drugs that can rapidly improve cognitive function on primary hippocampal neurons in mice

group Cell viability (%) P value blank group 100.00±0.00 control group 70.53±0.63 P<0.01 Example group 109.00±0.63 P<0.001

The compound provided in the present invention with rapid improvement of cognitive function can significantly increase the time of mice in the new arm in the 30min test after administration, improve the spatial memory of AD mice, and can rapidly enhance the intracellular calcium ion level at the same time, ATP levels in hippocampal neurons and long-term potentiation (LTP) in the hippocampus of mice, thereby rapidly increasing synaptic transmission in the hippocampus.

The above experiments proved that compared with the control group and the model group, the treatment group showed good and rapid improvement of cognitive function and spatial memory ability, and at the same time, it could rapidly enhance the ATP level and neuron activity of neurons in the hippocampus. In vitro experiments have shown that drugs that rapidly improve cognitive function have certain neuroprotective effects. To sum up, the compound of the present invention can be used as a compound for rapidly improving cognition and memory ability, preventing and treating Alzheimer's disease.

Each embodiment in this specification is described in a progressive manner, each embodiment focuses on the difference from other embodiments, and the same and similar parts of each embodiment can be referred to each other. As for the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and for the related information, please refer to the description of the method part.

The above description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the 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 invention. Therefore, the present invention will not 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 (2)

1. A compound having a rapid cognitive improvement, characterized by the chemical structure:

2. use of a compound according to claim 1 for the manufacture of a medicament for improving cognition.

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